2019 Vol. 35, No. 3
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2019, 35(3): 161-162.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.001
Abstract
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Abstract:
Objective A series of pathophysiological changes occur in a body with burn, among which the pathophysiological changes in organs are rather insidious but have far-reaching effects. At present, in clinical practice, we mainly adopt symptomatic or alternative treatment to deal with organ dysfunction, lacking systematic intervention measures based on clear and perfect theoretical data. Theme of the key article of this issue is the injury and protection of organs in the early stage after burns. The collected articles show the current status and frontiers of the research on this topic in China, including the retrospective analysis of clinical data, exploration of cell-level injury mechanism, and experimental treatment scheme in animal models. However, until now, few instructive or inspiring articles have been published. I appeal more colleagues could focus on this, explore more, research more, and share more.
Objective A series of pathophysiological changes occur in a body with burn, among which the pathophysiological changes in organs are rather insidious but have far-reaching effects. At present, in clinical practice, we mainly adopt symptomatic or alternative treatment to deal with organ dysfunction, lacking systematic intervention measures based on clear and perfect theoretical data. Theme of the key article of this issue is the injury and protection of organs in the early stage after burns. The collected articles show the current status and frontiers of the research on this topic in China, including the retrospective analysis of clinical data, exploration of cell-level injury mechanism, and experimental treatment scheme in animal models. However, until now, few instructive or inspiring articles have been published. I appeal more colleagues could focus on this, explore more, research more, and share more.
Wang Wensheng,
Xiang Fei,
Song Huapei,
Zhang Can,
Zhang Bingqian,
Lyu Yanling,
Yuan Hongping,
Hu Gaozhong,
Huang Yuesheng
2019, 35(3): 163-168.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.002
Abstract:
Objective To analyze the clinical characteristics of early organ injury in elderly patients with severe burns and the effects on the prognosis of patients. Methods From January 2010 to August 2018, 62 patients with severe burns (43 men and 19 women, aged from 60 to 89 years at the time of admission) who were hospitalized in the Institute of Burn Research of the First Affiliated Hospital of Army Medical University (the Third Military Medical University, hereinafter referred to as the author′s affiliation), meeting the inclusion criteria, were included in elderly (E) group, and 124 patients with severe burns (86 men and 38 women, aged from 18 to 59 years at the time of admission) at the same term were included in young and middle-aged (YM) group. Treatment of patients in the 2 groups followed the conventional procedures of the author′s affiliation. The following data of patients in the 2 groups were retrospectively analyzed. (1) Fluid replacement volume and urine volume within the first and second post injury hour (PIH) 24 were recorded. The levels of hemoglobin, haematocrit, and blood lactic acid at admission, PIH 24 and 48 were recorded. (2) The creatine kinase isozyme-MB (CK-MB), total bilirubin, blood creatinine, oxygenation index, and blood platelet count at admission, at shock stage, and on post injury day (PID) 3 to 7 were collected. (3) The days of seriously or critically ill and deaths were recorded. Data were processed with chi-square test, groupt test, Mann-Whitney U test, analysis of variance for repeated measurement, and Bonferroni correction.
Results (1) There were no statistically significant differences in fluid replacement volume within the first and second PIH 24, and urine volume within the second PIH 24 between patients in the 2 groups (t =0.351, 1.307, 1.110, P >0.05). The urine volume of patients in group E within the first PIH 24 was significantly less than that in group YM (t =5.628, P <0.05). There were no statistically significant differences in levels of hemoglobin (t =0.011, 1.075, 0.239), haematocrit (t =0, 0.033, 0.199), and blood lactic acid (t =0.017, 1.002, 0.739) at admission, PIH 24 and 48 between patients in the 2 groups (P >0.05). (2) There were no statistically significant differences in levels of CK-MB at admission and on PID 3 to 7 between patients in the 2 groups (t =0.069, 0.001, P >0.05). The level of CK-MB of patients in group E at shock stage was significantly higher than that in group YM (t =4.017, P <0.05). There were no statistically significant differences in levels of total bilirubin at admission and on PID 3 to 7 between patients in the 2 groups (t =0.227, 0.002, P >0.05). However, the level of total bilirubin of patients in group E at shock stage was significantly higher than that in group YM (t =6.485, P <0.05). The levels of blood creatinine of patients in group E at admission and shock stage were significantly higher than those in group YM (t =4.226, 12.299, P <0.05 or P <0.01), while there was no statistically significant difference between them on PID 3 to 7 (t =0.693, P >0.05). The oxygenation indexes of patients in group E at admission and shock stage and on PID 3 to 7 [(371±16), (263±16), and (228±18) mmHg (1 mmHg=0.133 kPa)] were lower than (420±13), (327±13), and (281±17) mmHg of patients in group YM, respectively (t =5.650, 9.782, 4.856, P <0.05 or P <0.01). There were no statistically significant differences in levels of blood platelet count at admission and shock stage between patients in the 2 groups (t =0.038, 0.588, P >0.05), while the level of blood platelet count of patients in group E on PID 3 to 7 was significantly lower than that in group YM (t =6.636, P <0.05). (3) The days of seriously or critically ill and death rate of patients in group E were respectively longer or higher than those in group YM (Z =-2.303, χ 2=13.676, P <0.05 or P <0.01).
Conclusions In the case of the same tissue perfusion at shock stage, injuries in heart, liver, kidney, lung, and coagulation system in elderly patients with severe burns are more obvious than those in young and middle-aged patients, with more severe illness and higher mortality.
Objective To analyze the clinical characteristics of early organ injury in elderly patients with severe burns and the effects on the prognosis of patients. Methods From January 2010 to August 2018, 62 patients with severe burns (43 men and 19 women, aged from 60 to 89 years at the time of admission) who were hospitalized in the Institute of Burn Research of the First Affiliated Hospital of Army Medical University (the Third Military Medical University, hereinafter referred to as the author′s affiliation), meeting the inclusion criteria, were included in elderly (E) group, and 124 patients with severe burns (86 men and 38 women, aged from 18 to 59 years at the time of admission) at the same term were included in young and middle-aged (YM) group. Treatment of patients in the 2 groups followed the conventional procedures of the author′s affiliation. The following data of patients in the 2 groups were retrospectively analyzed. (1) Fluid replacement volume and urine volume within the first and second post injury hour (PIH) 24 were recorded. The levels of hemoglobin, haematocrit, and blood lactic acid at admission, PIH 24 and 48 were recorded. (2) The creatine kinase isozyme-MB (CK-MB), total bilirubin, blood creatinine, oxygenation index, and blood platelet count at admission, at shock stage, and on post injury day (PID) 3 to 7 were collected. (3) The days of seriously or critically ill and deaths were recorded. Data were processed with chi-square test, group
Lin Jiezhi,
Yi Ruofan,
Zhang Xingyue,
Jia Jiezhi,
Zhang Qiong,
Cui Lin,
Yang Lei,
Ye Jingying,
Zhang Dongxia,
Lyu Yanling,
Huang Yuesheng
2019, 35(3): 169-178.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.003
Abstract:
Objective To investigate the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes cultured in vitro. Methods The hearts of 20 C57BL/6 mice aged 1-2 days no matter male or female were isolated to culture primary cardiomyocytes which were used in the following experiments. (1) The cells were divided into 5 groups according to the random number table (the same grouping method below), i. e., normal control group and sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups. The cells in normal control group were routinely cultured for 54.0 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), and the cells in sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups were firstly regularly cultured for 53.5, 53.0, 51.0, 48.0 h and then cultured with replaced sugar-free serum-free medium for 0.5, 1.0, 3.0, and 6.0 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ), autophagy-related protein 5, and adenosine triphosphatase V1 region E1 subunit (ATP6V1E1) were detected by Western blotting. (2) The cells were divided into normal control group and sugar-free serum-free 3.0 h group. The cells in corresponding groups were treated the same as those in experiment (1), and the cell lysosomal acidification level was observed and detected under a laser scanning confocal microscope. (3) Two batches of cells were grouped and treated the same as those in experiment (1). The protein expression of human antigen R in the whole protein of cells of one batch and its protein expression in the cytoplasm and nucleus protein of cells of the other batch were detected by Western blotting. (4) The cells were divided into normal control group, simple control small interfering RNA (siRNA) group, simple human antigen R-siRNA1 (HuR-siRNA1) group, simple HuR-siRNA2 group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group. After 48 hours of regular culture, the cells in simple control siRNA group and sugar-free serum-free+ control siRNA group were transfected with negative control siRNA for 6 h, the cells in simple HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA1 group were transfected with HuR-siRNA1 for 6 h, and the cells in simple HuR-siRNA2 group and sugar-free serum-free+ HuR-siRNA2 group were transfected with HuR-siRNA2 for 6 h. Hereafter, the cells in these 8 groups were continuously cultured for 48 h with regular conditon, and then the cells in normal control group and each simple siRNA-treated group were replaced with DMEM/F12 medium, the cells in the other groups were replaced with sugar-free serum-free medium, and they were cultured for 3 h. The protein expression of human antigen R in the whole protein of cells was detected by Western blotting. (5) Two batches of cells were divided into sugar-free serum-free+ control siRNA group and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The distribution and expression of human antigen R in the cells of one batch were observed and detected by immunofluorescence method, and the lysosomal acidification level in the cells of the other batch was observed and detected under a laser scanning confocal microscope. (6) Three batches of cells were divided into sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group, and the cells in corresponding groups were treated the same as those in experiment (4). The protein expressions of cathepsin D in the whole protein of cells of one batch, human antigen R in the cytoplasm protein of cells of one batch, and ATP6V1E1 in the whole protein of cells of the other batch were detected by Western blotting. (7) The cells were divided into normal control group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The mRNA expression of ATP6V1E1 in cells was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The sample number of each experiment was 3. Data were processed with independent datat test, one-way analysis of variance, least significant difference t test, and Bonferroni correction.
Results (1) Compared with those of normal control group, the protein expressions of LC3Ⅱ and ATP6V1E1 in the whole protein of cells of sugar-free serum-free 1.0, 3.0, and 6.0 h groups were significantly increased (t =12.16, 4.05, 4.82, 11.64, 3.29, 8.37, P <0.05 or P <0.01). Compared with that of normal control group, the protein expression of autophagy-related protein 5 in the whole protein of cells of sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups was significantly increased (t =6.88, 10.56, 5.76, 9.91, P <0.05 or P <0.01). (2) Compared with 1.03±0.08 of normal control group, the lysosomal acidification level in the cells of sugar-free serum-free 3.0 group (2.92±0.30) was significantly increased (t =6.01, P <0.01). (3) There was no statistically significant difference in the overall comparison of protein expression of human antigen R in the whole protein of cells among the 5 groups (F =1.09, P >0.05). Compared with that of normal control group, the protein expression of human antigen R in the cytoplasm protein of cells was significantly increased in sugar-free serum-free 1.0, 3.0, and 6.0 h groups (t =43.05, 11.07, 5.39, P <0.05 or P <0.01), while the protein expression of human antigen R in the nucleus protein of cells was significantly decreased in sugar-free serum-free 3.0 and 6.0 h groups (t =11.18, 12.71, P <0.01). (4) Compared with that of simple control siRNA group, the protein expression of human antigen R in the whole protein of cells of simple HuR-siRNA1 group and simple HuR-siRNA2 group was significantly decreased (t =4.82, 4.44, P <0.05). Compared with that of sugar-free serum-free+ control siRNA group, the protein expression of human antigen R in the whole protein of cells of sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group was significantly decreased (t =4.39, 6.27, P <0.05). (5) Compared with those of sugar-free serum-free+ control siRNA group, the distribution of human antigen R in the cytoplasm of cells and its expression level were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group (t =10.13, P <0.01). Compared with 1.00±0.06 of sugar-free serum-free+ control siRNA group, the lysosomal acidification level (0.73±0.06) in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t =3.28, P <0.01). (6) Compared with those of sugar-free serum-free+ control siRNA group, the protein expressions of cathepsin D in the whole protein of cells, human antigen R in the cytoplasm protein of cells, and ATP6V1E1 in the whole protein of cells were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group (t =4.16, 3.99, 4.81, 5.07, 11.68, 12.97, P <0.05 or P <0.01). (7) Compared with that of normal control group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free 3.0 h group was significantly increased (t =5.51, P <0.05). Compared with that of sugar-free serum-free+ control siRNA group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t =5.97, P <0.05).
Conclusions After sugar-free serum-free treatment in vitro, the autophagy in mouse primary cardiomyocytes is activated, the lysosomal acidification is enhanced, and the expression of human antigen R in cytoplasm is increased. Human antigen R function is activated and involved in maintaining lysosomal acidification during autophagy in mouse cardiomyocytes.
Objective To investigate the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes cultured in vitro. Methods The hearts of 20 C57BL/6 mice aged 1-2 days no matter male or female were isolated to culture primary cardiomyocytes which were used in the following experiments. (1) The cells were divided into 5 groups according to the random number table (the same grouping method below), i. e., normal control group and sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups. The cells in normal control group were routinely cultured for 54.0 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), and the cells in sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups were firstly regularly cultured for 53.5, 53.0, 51.0, 48.0 h and then cultured with replaced sugar-free serum-free medium for 0.5, 1.0, 3.0, and 6.0 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ), autophagy-related protein 5, and adenosine triphosphatase V1 region E1 subunit (ATP6V1E1) were detected by Western blotting. (2) The cells were divided into normal control group and sugar-free serum-free 3.0 h group. The cells in corresponding groups were treated the same as those in experiment (1), and the cell lysosomal acidification level was observed and detected under a laser scanning confocal microscope. (3) Two batches of cells were grouped and treated the same as those in experiment (1). The protein expression of human antigen R in the whole protein of cells of one batch and its protein expression in the cytoplasm and nucleus protein of cells of the other batch were detected by Western blotting. (4) The cells were divided into normal control group, simple control small interfering RNA (siRNA) group, simple human antigen R-siRNA1 (HuR-siRNA1) group, simple HuR-siRNA2 group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group. After 48 hours of regular culture, the cells in simple control siRNA group and sugar-free serum-free+ control siRNA group were transfected with negative control siRNA for 6 h, the cells in simple HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA1 group were transfected with HuR-siRNA1 for 6 h, and the cells in simple HuR-siRNA2 group and sugar-free serum-free+ HuR-siRNA2 group were transfected with HuR-siRNA2 for 6 h. Hereafter, the cells in these 8 groups were continuously cultured for 48 h with regular conditon, and then the cells in normal control group and each simple siRNA-treated group were replaced with DMEM/F12 medium, the cells in the other groups were replaced with sugar-free serum-free medium, and they were cultured for 3 h. The protein expression of human antigen R in the whole protein of cells was detected by Western blotting. (5) Two batches of cells were divided into sugar-free serum-free+ control siRNA group and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The distribution and expression of human antigen R in the cells of one batch were observed and detected by immunofluorescence method, and the lysosomal acidification level in the cells of the other batch was observed and detected under a laser scanning confocal microscope. (6) Three batches of cells were divided into sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group, and the cells in corresponding groups were treated the same as those in experiment (4). The protein expressions of cathepsin D in the whole protein of cells of one batch, human antigen R in the cytoplasm protein of cells of one batch, and ATP6V1E1 in the whole protein of cells of the other batch were detected by Western blotting. (7) The cells were divided into normal control group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The mRNA expression of ATP6V1E1 in cells was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The sample number of each experiment was 3. Data were processed with independent data
2019, 35(3): 179-185.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.004
Abstract:
Objective To explore the effects of different doses of dopamine on organ function of rats at early stage of severe scald. Methods Thirty-two male Wistar rats aged 8 to 12 weeks were divided into sham injury (SI) group, simple resuscitation (SR) group, small dose (SD) group, and moderate dose (MD) group according to the random number table, with 8 rats in each group. After rats in the 4 groups were performed cardiac catheterization, rats in group SI were sham injured on the back by immersing in 37 ℃ warm water for 18 s, and rats in the other 3 groups were inflicted with 30% total body surface area (TBSA) full-thickness scald on the back by immersing in 97 ℃ hot water for 18 s. Rats in group SI were not treated after the injury, while rats in the other 3 groups were performed fluid resuscitation for 24 h through jugular vein catheter with micro syringe pump according to the Parkland formula. They were given 4.0 mL·kg-1·% TBSA-1 normal saline during the first 24 h, of which they were given half of the total amount for the first 8 h, and they were given half of the total amount for the second and third 8 h. Rats in group SR were infused normal saline only, while rats in group SD and group MD were infused normal saline+ 1.25 μg·kg-1·min-1dopamine and normal saline+ 6.00 μg·kg-1·min-1 dopamine respectively. Volume of 0.5 mL venous blood of all rats were taken through the cardiac catheter with serum separated at post injury hour (PIH) 1, 3, 6, 12, and 24. Serum content of cardiac troponin I (cTnI) was determined by enzyme-linked immunosorbent assay; serum content of diamine oxidase (DAO) was detected by ultraviolet spectrophotometer; serum content of β2-microglobulin (β2-MG) was determined by latex-enhanced immunoturbidimetric assay; serum content of total bile acid (TBA) was determined by enzyme colorimetry; serum content of lactic acid, malondialdehyde, and myeloperoxidase (MPO) was determined by ultraviolet spectrophotometer. Data were processed with analysis of variance for repeated measurement, one-way analysis of variance, least significant difference test, and Bonferroni correction. Results (1) At PIH 1, 3, 6, 12, and 24, serum content of cTnI of rats in group SR, group SD, and group MD [(2.69±0.19), (3.04±0.19), (4.96±0.25), (6.88±0.28), (4.75±0.31) μg/L, (2.70±0.14), (3.08±0.13), (5.06±0.19), (7.11±0.21), (4.89±0.16) μg/L, (2.18±0.14), (2.54±0.09), (3.97±0.14), (5.46±0.34), (3.32±0.33) μg/L] were higher than that in group SI [(1.70±0.08), (1.70±0.08), (1.69±0.11), (1.69±0.08), (1.70±0.08) μg/L,P <0.05], serum content of cTnI of rats in group SR and group SD was similar (P >0.05), and serum content of cTnI of rats in group MD was lower than that in group SR and group SD (P <0.05). (2) At PIH 1 to 24, serum content of DAO of rats in group SR, group SD, and group MD was higher than that in group SI (P <0.05), serum content of DAO of rats in group SR and group MD was similar (P >0.05), and serum content of DAO of rats in group SD was lower than that in group SR and group MD (P <0.05). (3) At PIH 1 to 24, serum content of β2-MG of rats in group SR, group SD, and group MD was higher than that in group SI (P <0.05), serum content of β2-MG of rats in group SR and group MD was similar (P >0.05), and serum content of β2-MG of rats in group SD was lower than that in group SR and group MD (P <0.05). (4) At PIH 1 to 24, serum content of TBA of rats in group SR, group SD, and group MD was similar (P >0.05) and higher than that in group SI (P <0.05). (5) At PIH 1 to 24, serum content of lactic acid of rats in group SR, group SD, and group MD was higher than that in group SI (P <0.05), serum content of lactic acid of rats in group SR and group MD was similar (P >0.05), and serum content of lactic acid of rats in group SD was lower than that in group SR and group MD (P <0.05). (6) At PIH 1 to 24, serum content of malondialdehyde and MPO of rats in group SR, group SD, and group MD was higher than that in group SI (P <0.05), serum content of malondialdehyde and MPO of rats in group SR and group MD was similar (P >0.05), and serum content of malondialdehyde and MPO of rats in group SD was significantly lower than that in group SR and group MD (P <0.05).
Conclusions With effective liquid recovery, dopamine of MD can improve early cardiac function of rats with severe scald, while dopamine of SD can alleviate tissue ischemia and hypoxia, reduce oxygen free radical damage in internal organs, and improve functions of intestine and kidney.
Objective To explore the effects of different doses of dopamine on organ function of rats at early stage of severe scald. Methods Thirty-two male Wistar rats aged 8 to 12 weeks were divided into sham injury (SI) group, simple resuscitation (SR) group, small dose (SD) group, and moderate dose (MD) group according to the random number table, with 8 rats in each group. After rats in the 4 groups were performed cardiac catheterization, rats in group SI were sham injured on the back by immersing in 37 ℃ warm water for 18 s, and rats in the other 3 groups were inflicted with 30% total body surface area (TBSA) full-thickness scald on the back by immersing in 97 ℃ hot water for 18 s. Rats in group SI were not treated after the injury, while rats in the other 3 groups were performed fluid resuscitation for 24 h through jugular vein catheter with micro syringe pump according to the Parkland formula. They were given 4.0 mL·kg-1·% TBSA-1 normal saline during the first 24 h, of which they were given half of the total amount for the first 8 h, and they were given half of the total amount for the second and third 8 h. Rats in group SR were infused normal saline only, while rats in group SD and group MD were infused normal saline+ 1.25 μg·kg-1·min-1dopamine and normal saline+ 6.00 μg·kg-1·min-1 dopamine respectively. Volume of 0.5 mL venous blood of all rats were taken through the cardiac catheter with serum separated at post injury hour (PIH) 1, 3, 6, 12, and 24. Serum content of cardiac troponin I (cTnI) was determined by enzyme-linked immunosorbent assay; serum content of diamine oxidase (DAO) was detected by ultraviolet spectrophotometer; serum content of β2-microglobulin (β2-MG) was determined by latex-enhanced immunoturbidimetric assay; serum content of total bile acid (TBA) was determined by enzyme colorimetry; serum content of lactic acid, malondialdehyde, and myeloperoxidase (MPO) was determined by ultraviolet spectrophotometer. Data were processed with analysis of variance for repeated measurement, one-way analysis of variance, least significant difference test, and Bonferroni correction. Results (1) At PIH 1, 3, 6, 12, and 24, serum content of cTnI of rats in group SR, group SD, and group MD [(2.69±0.19), (3.04±0.19), (4.96±0.25), (6.88±0.28), (4.75±0.31) μg/L, (2.70±0.14), (3.08±0.13), (5.06±0.19), (7.11±0.21), (4.89±0.16) μg/L, (2.18±0.14), (2.54±0.09), (3.97±0.14), (5.46±0.34), (3.32±0.33) μg/L] were higher than that in group SI [(1.70±0.08), (1.70±0.08), (1.69±0.11), (1.69±0.08), (1.70±0.08) μg/L,
Wei Jinyu,
Cui Lin,
Lin Jiezhi,
Zhang Qiong,
Yuan Hongping,
Xiang Fei,
Song Huapei,
Jia Jiezhi,
Lyu Yanling,
Zhang Dongxia,
Huang Yuesheng
2019, 35(3): 186-192.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.005
Abstract:
Objective To explore the effects of transient receptor potential vanilloid 1 (TRPV1) on autophagy in early hypoxic mouse cardiomyocytes and the mechanism in vitro. Methods The hearts of 120 C57BL/6 mice aged 1-2 days, no matter male or female, were isolated, and then primary cardiomyocytes were cultured and used for the following experiments, the random number table was used for grouping. (1) The cells were divided into normoxia group and hypoxia 3, 6, and 9 h groups, with one well in each group. The cells in normoxia group were routinely cultured (the same below), the cells in hypoxia 3, 6, and 9 h groups were treated with fetal bovine serum-free and glucose-free Dulbecco′ s modified Eagle medium under low oxygen condition in a volume fraction of 1% oxygen, 5% carbon dioxide, and 94% nitrogen for 3, 6, and 9 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, TRPV1 were determined with Western botting. (2) The cells were divided into normoxia group and hypoxia group, with two coverslips in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above. The positive expression of TRPV1 was detected by immunofluorescence assay. (3) The cells were divided into 4 groups, with one well in each group. The cells in simple hypoxia group were treated with hypoxia for 6 h as above, and the cells in hypoxia+ 0.1 μmol/L capsaicin group, hypoxia+ 1.0 μmol/L capsaicin group, and hypoxia+ 10.0 μmol/L capsaicin group were respectively treated with 0.1, 1.0, 10.0 μmol/L capsaicin for 30 min before hypoxia for 6 h. The protein expressions of LC3, Beclin-1, and TRPV1 were detected by Western blotting. (4) The cells were divided into 5 groups, with 5 wells in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ chloroquine group, hypoxia+ capsaicin group, and hypoxia+ capsaicin+ chloroquine group were treated with hypoxia for 6 h after being cultured with 50 μmol/L chloroquine, 10.0 μmol/L capsaicin, and 50 μmol/L chloroquine+ 10.0 μmol/L capsaicin for 30 min, respectively. Viability of cells was detected by cell counting kit 8 assay. (5) The cells were divided into simple hypoxia group and hypoxia+ 10.0 μmol/L capsaicin group, with one well in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ 10.0 μmol/L capsaicin group were treated with 10.0 μmol/L capsaicin for 30 minutes and then with hypoxia for 6 h. The protein expressions of lysosomal associated membrane protein 1 (LAMP-1) and LAMP-2 were detected by Western blotting. Each experiment was repeated for 3 or 5 times. Data were processed with one-way analysis of variance, least significant differencet test, and Bonferroni correction.
Results (1) Compared with those of normoxia group, the protein expressions of LC3, Beclin-1, and TRPV1 were significantly increased in cardiomyocytes of hypoxia 3, 6, and 9 h groups (t 3 h=4.891, 5.890, 4.928; t 6 h=9.790, 6.750, 10.590; t 9 h=6.948, 6.764, 5.049, P <0.05 or P <0.01), which of hypoxia 6 h group were the highest (1.08±0.05, 1.12±0.10, 0.953±0.071, respectively). (2) The density of TRPV1 in cell membrane and inside the cardiomyocytes in hypoxia group was significantly increased with lump-like distribution, and the expression of TRPV1 was higher than that in normoxia group. (3) Compared with those of simple hypoxia group, the protein expression of Beclin-1 in cardiomyocytes of hypoxia+ 0.1 μmol/L capsaicin group was increased (t =10.488, P <0.01), while the protein expressions of LC3 and TRPV1 were increased without statistically significant differences (t =4.372, 3.026, P >0.05); the protein expressions of LC3, TRPV1, and Beclin-1 in cardiomyocytes of hypoxia+ 1.0 μmol/L capsaicin group and hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (t =15.505, 5.773, 13.430; 20.915, 8.054, 16.384; P <0.05 or P <0.01), which of hypoxia+ 10.0 μmol/L capsaicin group were the highest (2.33±0.09, 1.34±0.07, 1.246±0.053, respectively). (4) Compared with 0.585±0.045 in normoxia group, the cardiomyocyte viability in hypoxia group was significantly decreased (0.471±0.037, t =4.365, P <0.05). Compared with that in hypoxia group, the cardiomyocyte viability in hypoxia+ chloroquine group was further decreased (0.350±0.023, t =6.216, P <0.01), while 0.564±0.047 in hypoxia+ capsaicin group was significantly increased (t =3.489, P <0.05). Compared with that in hypoxia+ chloroquine group, the cardiomyocyte viability in hypoxia+ capsaicin+ chloroquine group did not significantly change (0.364±0.050, t =0.545, P >0.05). (5) Compared with 0.99±0.04 and 0.54±0.04 in simple hypoxia group, the protein expressions of LAMP-1 and LAMP-2 in hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (1.49±0.06, 0.81±0.05, t =12.550, 7.442, P <0.01).
Conclusions TRPV1 can further promote the expression of autophagy-related proteins in hypoxic cardiomyocytes through autophagy-lysosomal pathway, enhance autophagy activity, and improve autophagic flow for alleviating early hypoxic cardiomyocyte injury.
Objective To explore the effects of transient receptor potential vanilloid 1 (TRPV1) on autophagy in early hypoxic mouse cardiomyocytes and the mechanism in vitro. Methods The hearts of 120 C57BL/6 mice aged 1-2 days, no matter male or female, were isolated, and then primary cardiomyocytes were cultured and used for the following experiments, the random number table was used for grouping. (1) The cells were divided into normoxia group and hypoxia 3, 6, and 9 h groups, with one well in each group. The cells in normoxia group were routinely cultured (the same below), the cells in hypoxia 3, 6, and 9 h groups were treated with fetal bovine serum-free and glucose-free Dulbecco′ s modified Eagle medium under low oxygen condition in a volume fraction of 1% oxygen, 5% carbon dioxide, and 94% nitrogen for 3, 6, and 9 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, TRPV1 were determined with Western botting. (2) The cells were divided into normoxia group and hypoxia group, with two coverslips in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above. The positive expression of TRPV1 was detected by immunofluorescence assay. (3) The cells were divided into 4 groups, with one well in each group. The cells in simple hypoxia group were treated with hypoxia for 6 h as above, and the cells in hypoxia+ 0.1 μmol/L capsaicin group, hypoxia+ 1.0 μmol/L capsaicin group, and hypoxia+ 10.0 μmol/L capsaicin group were respectively treated with 0.1, 1.0, 10.0 μmol/L capsaicin for 30 min before hypoxia for 6 h. The protein expressions of LC3, Beclin-1, and TRPV1 were detected by Western blotting. (4) The cells were divided into 5 groups, with 5 wells in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ chloroquine group, hypoxia+ capsaicin group, and hypoxia+ capsaicin+ chloroquine group were treated with hypoxia for 6 h after being cultured with 50 μmol/L chloroquine, 10.0 μmol/L capsaicin, and 50 μmol/L chloroquine+ 10.0 μmol/L capsaicin for 30 min, respectively. Viability of cells was detected by cell counting kit 8 assay. (5) The cells were divided into simple hypoxia group and hypoxia+ 10.0 μmol/L capsaicin group, with one well in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ 10.0 μmol/L capsaicin group were treated with 10.0 μmol/L capsaicin for 30 minutes and then with hypoxia for 6 h. The protein expressions of lysosomal associated membrane protein 1 (LAMP-1) and LAMP-2 were detected by Western blotting. Each experiment was repeated for 3 or 5 times. Data were processed with one-way analysis of variance, least significant difference
2019, 35(3): 193-197.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.006
Abstract:
Objective To explore the effects of transcutaneous electrical acupoint stimulation (TEAS) on pain of patients in expansion process of skin soft tissue dilator on forehead by water injection. Methods From June 2016 to June 2017, 100 patients in expansion process of skin soft tissue dilator on forehead by water injection meeting the inclusion criteria were admitted to Outpatient Department of Orthopedic Surgery of Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine. There were 43 men and 57 women among the patients, aged 27 to 55 years, and the prospective randomized controlled study was performed on them. The patients were divided into TEAS nursing group and routine nursing group according to the random number table, with 50 patients in each group. Patients in routine nursing group were performed with routine nursing in every water injection in outpatient department and 2 days later. On the basis of routine nursing, patients in TEAS nursing group were performed with TEAS treatment by responsible nurses in each water injection in outpatient department. The Shangxing, Diwei, and Hegu points were positioned accurately, and electrical stimulation was performed on the 3 points simultaneously by pulse acupuncture treatment instrument, with 30 minutes each time. Two days after every water injection of outpatient department, TEAS nursing was performed at home by patients and their family members under remote guidance of the responsible nurses, with 2 times each day and 30 minutes each time. Besides, follow-up was done by phone by the responsible nurses everyday. The nursing of patients in the 2 groups lasted the whole expansion process. After the expansion process, the overall pain degree and the most severe pain degree of patients during expansion process were scored by numerical rating scale, and the overall comfort degree and its dimensions of patients during expansion process were scored by the responsible nurses every day by simplified Comfort Status Scale. Data were processed with independent samplet test and chi-square test.
Results During expansion process, the overall pain score degree and the most severe pain degree score of patients in TEAS nursing group were (5.4±1.2) and (6.5±1.0) points, which were significantly lower than (6.1±1.3) and (7.5±1.4) points of patients in routine nursing group (t =-2.62, -4.00, P <0.05 or P <0.01). During expansion process, the physiological dimension, sociocultural dimension, psychological spirit dimension, environmental dimension, and total score of the overall comfort degree of patients in TEAS nursing group were (9.6±2.9), (20.1±2.8), (29.1±1.9), (22±3), and (80±6) points, significantly higher than (5.7±2.1), (16.8±2.8), (26.0±2.8), (21±4), and (69±8) points of patients in routine nursing group (t =8.03, 6.35, 7.60, 2.11, 10.64, P <0.05 or P <0.01).
Conclusions TEAS with appropriate intensity, frequency, and duration can alleviate the pain of patients during expansion process of skin soft tissue dilator on forehead by water injection and improve their comfort degree.
Objective To explore the effects of transcutaneous electrical acupoint stimulation (TEAS) on pain of patients in expansion process of skin soft tissue dilator on forehead by water injection. Methods From June 2016 to June 2017, 100 patients in expansion process of skin soft tissue dilator on forehead by water injection meeting the inclusion criteria were admitted to Outpatient Department of Orthopedic Surgery of Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine. There were 43 men and 57 women among the patients, aged 27 to 55 years, and the prospective randomized controlled study was performed on them. The patients were divided into TEAS nursing group and routine nursing group according to the random number table, with 50 patients in each group. Patients in routine nursing group were performed with routine nursing in every water injection in outpatient department and 2 days later. On the basis of routine nursing, patients in TEAS nursing group were performed with TEAS treatment by responsible nurses in each water injection in outpatient department. The Shangxing, Diwei, and Hegu points were positioned accurately, and electrical stimulation was performed on the 3 points simultaneously by pulse acupuncture treatment instrument, with 30 minutes each time. Two days after every water injection of outpatient department, TEAS nursing was performed at home by patients and their family members under remote guidance of the responsible nurses, with 2 times each day and 30 minutes each time. Besides, follow-up was done by phone by the responsible nurses everyday. The nursing of patients in the 2 groups lasted the whole expansion process. After the expansion process, the overall pain degree and the most severe pain degree of patients during expansion process were scored by numerical rating scale, and the overall comfort degree and its dimensions of patients during expansion process were scored by the responsible nurses every day by simplified Comfort Status Scale. Data were processed with independent sample
2019, 35(3): 198-204.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.007
Abstract:
Objective To assess the cognitive level of first aid knowledge regarding the small area burn among the child caregivers in Shanghai and improve the level of first aid for small area burn in children. Methods From November 2017 to March 2018, 7 municipal districts in Shanghai were selected according to the random number table, from which 2 750 students of 4 nurseries, 5 kindergartens, 6 primary schools, and 2 junior middle schools were selected by adopting the convenience sampling method. Each student was limited to one caregiver as the research object. A cross-sectional survey was conducted on the cognitive level of first aid knowledge regarding small area burn among the caregivers with self-designed questionnaire through WeChat and Tencent QQ. The age, burn experience, and scarring after burns in children, the prevalence rate of burn in children of different age groups, the educational background of caregivers and their social relationship with their children, and the measures taken by caregivers firstly after small area burn occurred among their children were recorded. The choices of applying the folk prescription drugs to the wounds of their children made by caregivers and those with different educational backgrounds were recorded. The choices of applying daily necessities to the wound of their children made by caregivers were recorded. The caregivers′ knowledge of standard first aid measures for small area burn, and the knowledge of caregivers with different educational backgrounds of all standard first aid measures for small area burn were recorded. The caregivers′ choices of hospitals for treatment the first time, and the choices of going to the Grade Ⅲ Level A hospital with burn specialty for treatment made by caregivers with different knowledge levels about first aid measures for small area burn and those by caregivers whose children did or didn′t have burn experience were recorded. The caregivers′ choices of different types of medical institutions with burn specialty or specialized in burn treatment, and choices of going to burn department of comprehensive Grade Ⅲ Level A hospital for treatment made by caregivers with different knowledge levels about first aid measures for small area burn were recorded. Data were processed with Pearson chi-square test and partitions of chi-square test. Results The effective recovery rate of questionnaire was 99.0% (2 723/2 750). The ages of children were mainly 6-11 years [64.7% (1 762/2 723)]The prevalence of burn in children was 19.4% (527/2 723). There was no statistically significant difference in the overall comparison of burn prevalence of children among the age groups (χ 2=1.424, P >0.05). The percentage of scar formation after burn in children was 27.3% (144/527). The education backgrounds of caregivers were mainly undergraduate [40.2% (1 094/2 723)], and their social relationships with children were mainly children′s mothers [74.6% (2 030/2 723)]. Assuming that their children suffered from minor burns, the measures firstly taken by 74.0% (2 016/2 723) of the caregivers was to immediately access cool running water and remove clothing on the wound of children. Totally 19.2% (523/2 723) of the caregivers chose to apply folk prescription drugs for their burn children by themselves, and the percentage of caregivers with education background of junior middle school choosing to apply folk prescription drugs for their burn children by themselves was significantly higher than that of caregivers with education background of junior college, undergraduate, or graduate (χ 2=18.502, 20.642, 13.319, P <0.05). Totally 49.2% (1 340/2 723) of caregivers chose to daub many kinds of daily necessities for their burn children by themselves. Totally 39.2% (1 068/2 723) of caregivers knew all standard first aid measures for small area burn, the percentage of caregivers with education background of undergraduate knowing all standard first aid measures for small area burn was significantly higher than that of caregivers with education background of senior high school and secondary specialized school (χ 2=11.234, P <0.05). Assuming that their children suffered from minor burns, 39.0% (1 063/2 723) of the caregivers chose to go to the nearest hospital for treatment the first time, the percentage of caregivers who knew all standard first aid measures for small area burn choosing to go to Grade Ⅲ Level A hospital with burn specialty for treatment the first time was similar with that of caregivers who did not know/did not fully know (χ 2=3.528, P >0.05), and the percentage of caregivers whose children had burn experience choosing to go to Grade Ⅲ Level A hospital with burn specialty for treatment in the first time was similar with that of caregivers whose children didn′t have burn experience (χ 2=3.521, P >0.05). Among all medical institutions with burn specialty or specialized in burn treatment, 28.0% (762/2 723) of the caregivers chose to go to comprehensive Grade Ⅲ Level A hospital for treatment, and the percentage of caregivers who knew all standard first aid measures for small area burn choosing to go to comprehensive Grade Ⅲ Level A hospital for treatment was significantly higher than that of caregivers who did not know/did not fully know (χ 2=4.890, P <0.05).
Conclusions The caregivers of children are mainly children′s mothers with education background of undergraduate in Shanghai, and caregivers′ cognitive levels of first aid knowledge regarding the small area burn are low. Only a few caregivers know all standard first aid measures for small area burn, and there are still some caregivers who have the wrong idea of applying folk prescription drugs or daily necessities for children by themselves. The publicity and education of basic first aid knowledge of burn should be strengthened through various channels such as burn simulation exercise and network, and caregivers should be guided to take their children to hospitals with burn specialty for treatment after occurrence of burn in children, so as to obtain more professional medical treatment.
Objective To assess the cognitive level of first aid knowledge regarding the small area burn among the child caregivers in Shanghai and improve the level of first aid for small area burn in children. Methods From November 2017 to March 2018, 7 municipal districts in Shanghai were selected according to the random number table, from which 2 750 students of 4 nurseries, 5 kindergartens, 6 primary schools, and 2 junior middle schools were selected by adopting the convenience sampling method. Each student was limited to one caregiver as the research object. A cross-sectional survey was conducted on the cognitive level of first aid knowledge regarding small area burn among the caregivers with self-designed questionnaire through WeChat and Tencent QQ. The age, burn experience, and scarring after burns in children, the prevalence rate of burn in children of different age groups, the educational background of caregivers and their social relationship with their children, and the measures taken by caregivers firstly after small area burn occurred among their children were recorded. The choices of applying the folk prescription drugs to the wounds of their children made by caregivers and those with different educational backgrounds were recorded. The choices of applying daily necessities to the wound of their children made by caregivers were recorded. The caregivers′ knowledge of standard first aid measures for small area burn, and the knowledge of caregivers with different educational backgrounds of all standard first aid measures for small area burn were recorded. The caregivers′ choices of hospitals for treatment the first time, and the choices of going to the Grade Ⅲ Level A hospital with burn specialty for treatment made by caregivers with different knowledge levels about first aid measures for small area burn and those by caregivers whose children did or didn′t have burn experience were recorded. The caregivers′ choices of different types of medical institutions with burn specialty or specialized in burn treatment, and choices of going to burn department of comprehensive Grade Ⅲ Level A hospital for treatment made by caregivers with different knowledge levels about first aid measures for small area burn were recorded. Data were processed with Pearson chi-square test and partitions of chi-square test. Results The effective recovery rate of questionnaire was 99.0% (2 723/2 750). The ages of children were mainly 6-11 years [64.7% (1 762/2 723)]The prevalence of burn in children was 19.4% (527/2 723). There was no statistically significant difference in the overall comparison of burn prevalence of children among the age groups (
Zhou Jian,
Wei Zairong,
Sun Guangfeng,
Jin Wenhu,
Chang Shusen,
Li Hai,
Nie Kaiyu,
Tang Xiujun,
Gong Feiyu
2019, 35(3): 205-208.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.008
Abstract:
Objective To investigate the effects of free mini-flap on tibial side of third toe on repairing skin and soft tissue defect of finger pulp at the end of finger. Methods From August 2013 to May 2017, 18 patients with skin and soft tissue defect of finger pulp at the end of finger were admitted to our unit, with 12 men and 6 women aged 16 to 54 years. As the skin and soft tissue defect sites, there were 3 cases of thumb, 8 cases of index finger, 4 cases of middle finger, and 3 cases of ring finger. The area of defects ranged from 2.0 cm×1.4 cm to 3.5 cm×2.4 cm. Free mini-flaps on tibial side of third toes were designed according to area and shape of defects, and the length and width of flaps were 0.1 to 0.2 cm longer than the length and width of the defects, respectively. The area of flaps ranged from 2.1 cm×1.5 cm to 3.7 cm×2.6 cm. The end-to-end anastomosis of subcutaneous veins of flaps and superficial veins of the finger-palm side or superficial dorsal digital vein, the end-to-end tension-free anastomosis of the base metatarsal arteries on tibial side of third toe and proper digital arteries of recipient finger were performed. Besides, anastomosis of base metatarsal nerve on tibial side of third toe and proper digital nerve of recipient finger was performed. The donor sites on feet were sutured directly or repaired with full-thickness skin grafts on medial upper leg of the same side. The survival of flaps after operation and the follow-up of patients were observed. Results All flaps survived well, with good blood supply. Among the 18 patients, 2 patients lost to follow-up, and 16 patients were followed up for 4 to 36 months. The shape and texture of flaps were good. After reconstruction, finger pulps at the end of finger were plump, with fingerprint. Function of the finger restored well, and the two-point discriminatory distances of flaps were 5 to 10 mm. The donor sites on feet of 14 patients healed after the operation, the other 2 patients had necrosis on edge and central area of skin grafts, and the necrotic area healed after dressing change. The skin graft areas on feet were wear-resistant, with slight damage to donor sites and did not influence shoes wearing and walking. Besides, patients did not feel uncomfortable. Conclusions Skin and soft tissue defects of finger pulp at the end of finger repaired by free mini-flaps on tibial side of third toe are with good shape and slight damage to donor sites, and the operation is simple. It is worthy of popularization and application in clinic.
Objective To investigate the effects of free mini-flap on tibial side of third toe on repairing skin and soft tissue defect of finger pulp at the end of finger. Methods From August 2013 to May 2017, 18 patients with skin and soft tissue defect of finger pulp at the end of finger were admitted to our unit, with 12 men and 6 women aged 16 to 54 years. As the skin and soft tissue defect sites, there were 3 cases of thumb, 8 cases of index finger, 4 cases of middle finger, and 3 cases of ring finger. The area of defects ranged from 2.0 cm×1.4 cm to 3.5 cm×2.4 cm. Free mini-flaps on tibial side of third toes were designed according to area and shape of defects, and the length and width of flaps were 0.1 to 0.2 cm longer than the length and width of the defects, respectively. The area of flaps ranged from 2.1 cm×1.5 cm to 3.7 cm×2.6 cm. The end-to-end anastomosis of subcutaneous veins of flaps and superficial veins of the finger-palm side or superficial dorsal digital vein, the end-to-end tension-free anastomosis of the base metatarsal arteries on tibial side of third toe and proper digital arteries of recipient finger were performed. Besides, anastomosis of base metatarsal nerve on tibial side of third toe and proper digital nerve of recipient finger was performed. The donor sites on feet were sutured directly or repaired with full-thickness skin grafts on medial upper leg of the same side. The survival of flaps after operation and the follow-up of patients were observed. Results All flaps survived well, with good blood supply. Among the 18 patients, 2 patients lost to follow-up, and 16 patients were followed up for 4 to 36 months. The shape and texture of flaps were good. After reconstruction, finger pulps at the end of finger were plump, with fingerprint. Function of the finger restored well, and the two-point discriminatory distances of flaps were 5 to 10 mm. The donor sites on feet of 14 patients healed after the operation, the other 2 patients had necrosis on edge and central area of skin grafts, and the necrotic area healed after dressing change. The skin graft areas on feet were wear-resistant, with slight damage to donor sites and did not influence shoes wearing and walking. Besides, patients did not feel uncomfortable. Conclusions Skin and soft tissue defects of finger pulp at the end of finger repaired by free mini-flaps on tibial side of third toe are with good shape and slight damage to donor sites, and the operation is simple. It is worthy of popularization and application in clinic.
Hu Delin,
Yu Youxin,
Liang Rong,
Zhou Shunying,
Duan Shengliang,
Jiang Zhiyong,
Meng Chengying,
Jiang Wei,
Wang Huan,
Sun Yexiang,
Fang Linsen
2019, 35(3): 209-217.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.009
Abstract:
Objective To investigate the regulation of hypoxia-inducible factor-1α (HIF-1α) on permeability of rat vascular endothelial cells and the mechanism. Methods Twelve male Sprague-Dawley rats aged 35 to 38 days were collected and vascular endothelial cells were separated and cultured. The morphology of cells was observed after 4 days of culture, and the following experiments were performed on the 2nd or 3rd passage of cells. (1) Rat vascular endothelial cells were collected and divided into blank control group, negative control group, HIF-1α interference sequence 1 group, HIF-1α interference sequence 2 group, and HIF-1α interference sequence 3 group according to the random number table (the same grouping method below), with 3 wells in each group. Cells in negative control group, HIF-1α interference sequence 1 group, HIF-1α interference sequence 2 group, and HIF-1α interference sequence 3 group were transfected with GV248 empty plasmid, recombinant plasmid respectively containing HIF-1α interference sequence 1, interference sequence 2, and interference sequence 3 with liposome 2000. Cells in blank control group were only transfected with liposome 2000. After transfection of 24 h, expression levels of HIF-1α mRNA and protein of cells in each group were respectively detected by reverse transcription real-time fluorescent quantitative polymerase chain reaction and Western blotting (the same detecting methods below) . The sequence with the highest interference efficiency was selected. (2) Another batch of rat vascular endothelial cells were collected and divided into blank control group, negative control group, and HIF-1α low expression group, with 3 wells in each group. Cells in blank control group were only transfected with liposome 2000, and cells in negative control group and HIF-1α low expression group were respectively transfected with GV248 empty plasmid and low expression HIF-1α recombinant plasmid selected in experiment (1) with liposome 2000. After 14 days of culture, the mRNA and protein expressions of HIF-1α in each group were detected. (3) Another batch of rat vascular endothelial cells were collected and divided into blank control group, negative control group, and HIF-1α high expression group, with 3 wells in each group. Cells in blank control group were transfected with liposome 2000, and cells in negative control group and HIF-1α high expression group were respectively transfected with GV230 empty plasmid and HIF-1α high expression recombinant plasmid with liposome 2000. After 14 days of culture, the mRNA and protein expressions of HIF-1α of cells in each group were detected. (4) After transfection of 24 h, cells of three groups in experiment (1) and three groups in experiment (2) were collected, and mRNA and protein expressions of myosin light chain kinase (MLCK), phosphorylated myosin light chain (p-MLC), and zonula occludens 1 (ZO-1) of cells were detected. Data were processed with one-way analysis of variance andt test.
Results After 4 days of culture, the cells were spindle-shaped, and rat vascular endothelial cells were successfully cultured. (1) The interference efficiencies of HIF-1α of cells in HIF-1α interference sequence 1 group, HIF-1α interference sequence 2 group, and HIF-1α interference sequence 3 group were 47.66%, 45.79%, and 62.62%, respectively, and the interference sequence 3 group had the highest interference efficiency. After transfection of 24 h, the mRNA and protein expression levels of HIF-1α of cells in interference sequence 3 group were significantly lower than those in blank control group (t =18.404, 9.140, P <0.01) and negative control group (t =15.099, 7.096, P <0.01). (2) After cultured for 14 days, the mRNA and protein expression levels of HIF-1α of cells in HIF-1α low expression group were significantly lower than those in blank control group (t =21.140, 5.440, P <0.01) and negative control group (t = 14.310, 5.210, P <0.01). (3) After cultured for 14 days, the mRNA and protein expression levels of HIF-1α of cells in HIF-1α high expression group were significantly higher than those in blank control group (t =19.160, 7.710, P <0.01) and negative control group (t = 19.890, 7.500, P <0.01). (4) After transfection of 24 h, the mRNA expression levels of MLCK and p-MLC of cells in HIF-1α low expression group were significantly lower than those in blank control group (t =2.709, 4.011, P <0.05 or P <0.01) and negative control group (t =2.373, 3.744, P <0.05 or P <0.01). The mRNA expression level of ZO-1 of cells in HIF-1α low expression group was significantly higher than that in blank control group and negative control group (t =4.285, 5.050, P <0.01). The mRNA expression levels of MLCK and p-MLC of cells in HIF-1α high expression group were significantly higher than those in blank control group (t =9.118, 11.313, P <0.01) and negative control group (t =9.073, 11.280, P <0.01). The mRNA expression level of ZO-1 of cells in HIF-1α high expression group was significantly lower than that in blank control group and negative control group (t =2.889, 2.640, P <0.05). (5) After transfection of 24 h, the protein expression levels of MLCK and p-MLC of cells in HIF-1α low expression group were significantly lower than those in blank control group (t =2.652, 3.983, P <0.05 or P <0.01) and negative control group (t =2.792, 4.065, P <0.05 or P <0.01). The protein expression of ZO-1 of cells in HIF-1α low expression group was significantly higher than that in blank control group and negative control group (t =3.881, 3.570, P <0.01). The protein expression levels of MLCK and p-MLC of cells in HIF-1α high expression group were 1.18±0.24 and 0.68±0.22, which were significantly higher than 0.41±0.21 and 0.35±0.14 in blank control group (t =5.011, 3.982, P <0.05 or P <0.01) and 0.43±0.20 and 0.36±0.12 in negative control group (t = 4.880, 3.862, P <0.05 or P <0.01). The protein expression level of ZO-1 of cells in HIF-1α high expression group was 0.08±0.06, which was significantly lower than 0.20±0.09 in blank control group and 0.19±0.09 in negative control group (t =4.178, 3.830, P <0.05 or P <0.01).
Conclusions HIF-1α up-regulates expressions of MLCK and p-MLC and down-regulates expression of ZO-1, thereby increasing the permeability of rat vascular endothelial cells.
Objective To investigate the regulation of hypoxia-inducible factor-1α (HIF-1α) on permeability of rat vascular endothelial cells and the mechanism. Methods Twelve male Sprague-Dawley rats aged 35 to 38 days were collected and vascular endothelial cells were separated and cultured. The morphology of cells was observed after 4 days of culture, and the following experiments were performed on the 2nd or 3rd passage of cells. (1) Rat vascular endothelial cells were collected and divided into blank control group, negative control group, HIF-1α interference sequence 1 group, HIF-1α interference sequence 2 group, and HIF-1α interference sequence 3 group according to the random number table (the same grouping method below), with 3 wells in each group. Cells in negative control group, HIF-1α interference sequence 1 group, HIF-1α interference sequence 2 group, and HIF-1α interference sequence 3 group were transfected with GV248 empty plasmid, recombinant plasmid respectively containing HIF-1α interference sequence 1, interference sequence 2, and interference sequence 3 with liposome 2000. Cells in blank control group were only transfected with liposome 2000. After transfection of 24 h, expression levels of HIF-1α mRNA and protein of cells in each group were respectively detected by reverse transcription real-time fluorescent quantitative polymerase chain reaction and Western blotting (the same detecting methods below) . The sequence with the highest interference efficiency was selected. (2) Another batch of rat vascular endothelial cells were collected and divided into blank control group, negative control group, and HIF-1α low expression group, with 3 wells in each group. Cells in blank control group were only transfected with liposome 2000, and cells in negative control group and HIF-1α low expression group were respectively transfected with GV248 empty plasmid and low expression HIF-1α recombinant plasmid selected in experiment (1) with liposome 2000. After 14 days of culture, the mRNA and protein expressions of HIF-1α in each group were detected. (3) Another batch of rat vascular endothelial cells were collected and divided into blank control group, negative control group, and HIF-1α high expression group, with 3 wells in each group. Cells in blank control group were transfected with liposome 2000, and cells in negative control group and HIF-1α high expression group were respectively transfected with GV230 empty plasmid and HIF-1α high expression recombinant plasmid with liposome 2000. After 14 days of culture, the mRNA and protein expressions of HIF-1α of cells in each group were detected. (4) After transfection of 24 h, cells of three groups in experiment (1) and three groups in experiment (2) were collected, and mRNA and protein expressions of myosin light chain kinase (MLCK), phosphorylated myosin light chain (p-MLC), and zonula occludens 1 (ZO-1) of cells were detected. Data were processed with one-way analysis of variance and
Jin Wenhu,
Chang Shusen,
Wei Zairong,
Li Hai,
Zhou Jian,
Chen Wei,
Sun Guangfeng,
Tang Xiujun,
Wang Bo
2019, 35(3): 218-220.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.010
Abstract:
Objective To explore the clinical effects of heel lateral flap in repair of skin and soft tissue defects at posterior heel region. Methods From September 2007 to April 2016, 24 patients (17 males and 7 females, aged 16-70 years) with skin and soft tissue defects at posterior heel region were admitted to our department. The size of skin and soft tissue defects after debridement ranged from 3.0 cm×2.0 cm to 5.0 cm×4.0 cm. The defects were repaired with heel lateral flaps, with size ranging from 3.5 cm×2.5 cm to 6.0 cm×5.0 cm. The flaps were transferred to the donor sites through the loose subcutaneous tunnel. The donor site was repaired by full-thickness skin graft collected from inguinal region. The survival of flaps and the follow-up of patients were observed. Results All flaps of 24 patients survived successfully. The recipient sites and donor sites were all healed. The patients all had follow-up of 6 to 24 months. At the last follow-up, the flaps were in good shape, with nearly normal color and soft texture. There were 6 cases of grade S3 sensation and 16 cases of grade S3+ sensation. The distance of two-point discrimination of flaps ranged from 6 to 11 mm. The lateral foot skin grafts healed well, and the skin of the lateral foot was numb in the range of 4.0 cm×2.0 cm to 9.0 cm×3.0 cm. Conclusions Heel lateral flap can not only repair the skin and soft tissue defects in the posterior region, but also reconstruct the sensory function of the posterior region. It is an ideal method to repair the skin and soft tissue defects in the posterior region.
Objective To explore the clinical effects of heel lateral flap in repair of skin and soft tissue defects at posterior heel region. Methods From September 2007 to April 2016, 24 patients (17 males and 7 females, aged 16-70 years) with skin and soft tissue defects at posterior heel region were admitted to our department. The size of skin and soft tissue defects after debridement ranged from 3.0 cm×2.0 cm to 5.0 cm×4.0 cm. The defects were repaired with heel lateral flaps, with size ranging from 3.5 cm×2.5 cm to 6.0 cm×5.0 cm. The flaps were transferred to the donor sites through the loose subcutaneous tunnel. The donor site was repaired by full-thickness skin graft collected from inguinal region. The survival of flaps and the follow-up of patients were observed. Results All flaps of 24 patients survived successfully. The recipient sites and donor sites were all healed. The patients all had follow-up of 6 to 24 months. At the last follow-up, the flaps were in good shape, with nearly normal color and soft texture. There were 6 cases of grade S3 sensation and 16 cases of grade S3+ sensation. The distance of two-point discrimination of flaps ranged from 6 to 11 mm. The lateral foot skin grafts healed well, and the skin of the lateral foot was numb in the range of 4.0 cm×2.0 cm to 9.0 cm×3.0 cm. Conclusions Heel lateral flap can not only repair the skin and soft tissue defects in the posterior region, but also reconstruct the sensory function of the posterior region. It is an ideal method to repair the skin and soft tissue defects in the posterior region.
2019, 35(3): 221-223.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.011
Abstract:
From June to November 2016, 5 patients with severe burns were admitted to our unit. Broad-spectrum antibiotic and fluconazole were used on patients as earlier empirical anti-infection therapy of bacteria and fungi. Seven to twenty-one days after injury, 5 patients developed fungal infection. Antifungal agents of caspofungin, voriconazole, and amphotericin B liposomewere were used according to the results of fungal culture, and the infected wounds were also treated with repeated debridement and dressing change. Multiple autologous skin grafts were performed after infection control of wounds. With the above antifungal infection treatment for 5 to 11 days, 2 patients′ condition tended to be stable, and no fungus was found in wound secretion after cultured for many times. The patients were discharged with wounds healed after 52 to 54 days′ hospital stay. Due to severe burns degree and or elder age, fungal infection aggravated and expanded to the trunk in the other 3 patients, then developed into burn sepsis, resulting in patients died of multiple organ failure secondary to sepsis.
From June to November 2016, 5 patients with severe burns were admitted to our unit. Broad-spectrum antibiotic and fluconazole were used on patients as earlier empirical anti-infection therapy of bacteria and fungi. Seven to twenty-one days after injury, 5 patients developed fungal infection. Antifungal agents of caspofungin, voriconazole, and amphotericin B liposomewere were used according to the results of fungal culture, and the infected wounds were also treated with repeated debridement and dressing change. Multiple autologous skin grafts were performed after infection control of wounds. With the above antifungal infection treatment for 5 to 11 days, 2 patients′ condition tended to be stable, and no fungus was found in wound secretion after cultured for many times. The patients were discharged with wounds healed after 52 to 54 days′ hospital stay. Due to severe burns degree and or elder age, fungal infection aggravated and expanded to the trunk in the other 3 patients, then developed into burn sepsis, resulting in patients died of multiple organ failure secondary to sepsis.
2019, 35(3): 224-227.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.012
Abstract:
On 17th June 2017, a 50 years old man with refractory gout was admitted in our hospital. During the treatment, he was accompanied by intermittent fever (39 to 40 ℃) of unknown origin for 60 days and gastrointestinal bleeding, with difficult wound repair. After comprehensive treatment of thorough debridement, vacuum sealing drainage, skin graft, skin flap repair, and drug administration, the patient was discharged fully recovered on post hospitalization day 104.
On 17th June 2017, a 50 years old man with refractory gout was admitted in our hospital. During the treatment, he was accompanied by intermittent fever (39 to 40 ℃) of unknown origin for 60 days and gastrointestinal bleeding, with difficult wound repair. After comprehensive treatment of thorough debridement, vacuum sealing drainage, skin graft, skin flap repair, and drug administration, the patient was discharged fully recovered on post hospitalization day 104.
2019, 35(3): 227-228.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.013
Abstract:
A 33 years old male patient who suffered from a flame burn of 88% total body surface area was admitted to our hospital on November 28th, 2016. During his hospitalization, we repeatedly performed central vein catheterization in internal jugular veins, subclavian veins, or femoral veins for fluid transfusion. We incidentally found bilateral internal jugular vein thrombosis by performing a point-of-care ultrasound examination before catheterizing sometime. We treated the patient by avoiding catheterization in the affected internal jugular veins, anticoagulating with low molecular weight heparin, closing the wounds with skin autografting, and guiding the patient to practice functional exercise. The thrombus disappeared in the end. The patient was cured and discharged 3 months post burn.
A 33 years old male patient who suffered from a flame burn of 88% total body surface area was admitted to our hospital on November 28th, 2016. During his hospitalization, we repeatedly performed central vein catheterization in internal jugular veins, subclavian veins, or femoral veins for fluid transfusion. We incidentally found bilateral internal jugular vein thrombosis by performing a point-of-care ultrasound examination before catheterizing sometime. We treated the patient by avoiding catheterization in the affected internal jugular veins, anticoagulating with low molecular weight heparin, closing the wounds with skin autografting, and guiding the patient to practice functional exercise. The thrombus disappeared in the end. The patient was cured and discharged 3 months post burn.
2019, 35(3): 229-232.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.014
Abstract:
Deepening of wounds not only prolongs the wound repair time, increases the chance of scar formation after healing, but also is one of the important causes of death in severe burn patients. How to prevent wound deepening is a clinical problem in the treatment of burns. The mechanism of deepening burn wounds has not been fully elucidated, and the prevention and treatment measures in clinic need to be further explored. Based on the research results of the early deepening mechanism and prevention measures of burn wounds at home and abroad, this paper intends to summarize the three aspects including deepening mechanism, prevention measures, and research prospects of burn wounds.
Deepening of wounds not only prolongs the wound repair time, increases the chance of scar formation after healing, but also is one of the important causes of death in severe burn patients. How to prevent wound deepening is a clinical problem in the treatment of burns. The mechanism of deepening burn wounds has not been fully elucidated, and the prevention and treatment measures in clinic need to be further explored. Based on the research results of the early deepening mechanism and prevention measures of burn wounds at home and abroad, this paper intends to summarize the three aspects including deepening mechanism, prevention measures, and research prospects of burn wounds.
2019, 35(3): 233-236.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.015
Abstract:
Scars formed by various injuries can affect the appearance and psychology of patients. Therefore, more and more people pay attention to the prevention and treatment of scar. With the development of the autologous fat grafting, it has been gradually applied to the prevention and treatment of scar. At present, it has been confirmed by scholars that the autologous fat grafting could be an effective method to prevent and treat scar from basic research and clinical practice. At the same time, the deficiency of autologous fat grafting in the prevention and treatment of scar was also pointed out. This paper reviews the application, mechanism, and deficiency of autologous fat grafting in scar prevention and treatment.
Scars formed by various injuries can affect the appearance and psychology of patients. Therefore, more and more people pay attention to the prevention and treatment of scar. With the development of the autologous fat grafting, it has been gradually applied to the prevention and treatment of scar. At present, it has been confirmed by scholars that the autologous fat grafting could be an effective method to prevent and treat scar from basic research and clinical practice. At the same time, the deficiency of autologous fat grafting in the prevention and treatment of scar was also pointed out. This paper reviews the application, mechanism, and deficiency of autologous fat grafting in scar prevention and treatment.
2019, 35(3): 237-240.
doi: 10.3760/cma.j.issn.1009-2587.2019.03.016
Abstract:
Severe skin damage not only causes a mass of tissue defect, but also leads to the loss of various sensory functions. Tissue engineering skin provides a new way for high-quality wound repair, while there are still many problems in the recovery of sensory function, such as abnormality or loss of sensation of pain, touch, and temperature. Therefore, when tissue engineering skin is used to promote wound healing, regeneration and functional recovery of sensory nerve have attracted more and more attention. This article introduces the kind, distribution, regeneration, and factors influencing regeneration of sensory nerve in skin, and explores strategies in promoting regeneration of sensory nerve from dermal scaffold, seed cell, and neurturin of tissue engineering skin.
Severe skin damage not only causes a mass of tissue defect, but also leads to the loss of various sensory functions. Tissue engineering skin provides a new way for high-quality wound repair, while there are still many problems in the recovery of sensory function, such as abnormality or loss of sensation of pain, touch, and temperature. Therefore, when tissue engineering skin is used to promote wound healing, regeneration and functional recovery of sensory nerve have attracted more and more attention. This article introduces the kind, distribution, regeneration, and factors influencing regeneration of sensory nerve in skin, and explores strategies in promoting regeneration of sensory nerve from dermal scaffold, seed cell, and neurturin of tissue engineering skin.
