中华烧伤与创面修复杂志

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Objective To explore the mechanism of dendritic epidermal T lymphocytes (DETCs) in promoting healing of full-thickness skin defect wound on mice by regulating the proliferation and differentiation of epidermal stem cells (ESCs) in mice. Methods (1) Ten 8-week-old wild type (WT) male C57BL/6 mice (the same sex and kind below) were sacrificed to collect the skin of back for extracting DETCs to culture. Five WT and five 8-week-old T cell receptor (TCR) δ^-/^- mice were selected and enrolled in WT control group and TCR δ^-/^- control group, respectively. A full-thickness skin defect wound with diameter of 6 mm was made on both sides of spinal line on the back of mice without any treatment after injury. Another fifteen 8-week-old TCR δ^-/^- mice were selected and divided into phosphate buffer solution (PBS), DETC, and insulin-like growth factor-Ⅰ(IGF-Ⅰ) groups according to the random number table (the same grouping method below), with 5 mice in each group, and the same full-thickness skin defect wound was made on each mouse. Immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 μL sterile PBS , DETCs (cell concentration of 1×10⁶/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. The percentage of the residual wound area was calculated on post injury day (PID) 2, 4, 6, and 8. (2) Three 8-week-old WT mice were enrolled in WT control group and nine 8-week-old TCR δ^-/^- mice were divided into TCR δ^-/^- control group, PBS group, and DETC group, with 3 mice in each group. The full-thickness skin defect wound was made as in experiment (1) . On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin was detected by chemiluminescence imaging analyzer. (3) Three 8-week-old WT mice were enrolled in WT control group and six 8-week-old TCR δ^-/^- mice were divided into PBS and DETC groups, with 3 mice in each group, and the full-thickness skin defect wound was made as in experiment (1). On PID3, DETCs were extracted from the wound margin epidermis tissue to detect the percentage of DETCs expressing IGF-Ⅰ by flow cytometer. (4) The mice were taken as in experiment (2) and divided into WT control, PBS, DETC, and IGF-Ⅰ groups. A straight full-thickness skin defect incision with length of 3 cm was made in the direction of one inner ear. Mice in WT control group didn′t have any other treatment after injury, and immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 μL sterile PBS, DETCs (cell concentration of 1×10⁶/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. On PID 12, epidermis tissue of wound margin was collected, and immunofluorescence staining was performed to observe the number of keratin 15 positive cells. (5) The same mice were collected, grouped, and treated as in experiment (4). On PID12, the epidermis tissue of wound margin was collected and immunofluorescence staining was performed to observe the number of keratin 10 positive cells. (6) Twenty 3-day-old WT mice (the same below) were sacrificed to collect the whole skin, which was used to extract ESCs, with 5 mice detecting one index. The ESCs were divided into DETC co-culture group and control group, which were added with 1 mL DETCs (cell concentration of 1.25×10⁶/mL) and DETC medium, respectively. The percentage of 5-ethynyl-2′-deoxyuridine (EdU) positive cell on culture day (CD) 3, the percentages of CD49f⁺ CD71^- and keratin 14 positive cells on CD 5, and the percentage of keratin 10 positive cell on CD 10 in 2 groups were detected by flow cytometer. (7) Twenty mice were taken to extract ESCs, with 5 mice detecting one index. The ESCs were divided into control group and IGF-Ⅰ group, which were added with 1 mL sterile PBS and 10 ng/mL recombinant mice IGF-Ⅰ, respectively. The percentages of EdU positive cell, CD49f⁺ CD71^- cell, keratin10 positive cell, and keratin 14 positive cell were detected as in experiment (6). The sample in each group of experiments (6) and (7) was three. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and t test. Results (1) On PID 4, 6, and 8, the percentage of residual wound area in TCR δ^-/^- control group was significantly higher than that in WT control group (t=2.78, 3.39, 3.66, P<0.05 or P<0.01). The percentage of residual wound area in DETC group and IGF-Ⅰgroup on PID 4, 6, and 8 was apparently lower than that in PBS group (t=2.61, 3.21, 3.88, 2.84, 2.91, 2.49, P<0.05 or P<0.01). (2) On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in TCR δ^-/^- control group was significantly lower than that in WT control group (t=17.34, P<0.01). The protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in DETC group was significantly higher than that in PBS group (t=11.71, P<0.01). (3) On PID 3, the percentage of DETCs expressing IGF-Ⅰ in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group and DETC group (t=24.95, 27.23, P<0.01). (4) On PID 12, the number of keratin 15 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group, DETC group, and IGF-Ⅰ group (t=17.97, 11.95, 7.63, P<0.01). (5) The number of keratin 10 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly higher than that in WT control group, DETC group, and IGF-Ⅰ group (t=11.59, 9.51, 3.48, P<0.05 or P<0.01). (6) The percentages of EdU positive cells on CD 3, CD49f⁺ CD71^- cells on CD 5, and keratin 14 positive cells on CD 5 in DETC co-culture group were respectively (43.5±0.6)%, (66.5±0.5)%, (69.3±1.7)%, apparently higher than (32.3±1.3)%, (56.4±0.3)%, (54.9±1.3)% in control group (t=7.97, 17.10, 6.66, P<0.01). The percentage of keratin 10 positive cells on CD 10 in DETC co-culture group was (55.7±0.7)%, significantly lower than (67.1±1.2)% in control group (t=8.34, P<0.01). (7) The percentages of EdU positive cells on CD 3, CD49f⁺ CD71^- cells on CD 5, and keratin 14 positive cells on CD 5 in IGF-Ⅰ group were respectively (42.1±0.9)%, (81.1±1.3)%, (66.8±1.0)%, apparently higher than (32.4±0.7)%, (74.9±0.7)%, (52.0±1.9)% in control group (t=8.39, 4.24, 7.25, P<0.05 or P<0.01). The percentage of keratin 10 positive cells on CD 10 in IGF-Ⅰ group was (53.5±1.1)% , significantly lower than (58.2±0.3)% in control group (t=3.99, P<0.05). Conclusions DETCs can promote the proliferation and anti-apoptotic potential of ESCs and inhibit their differentiation into end-stage by secreting IGF-Ⅰ, thus promoting wound healing of full-thickness skin defects in mice.

Exploration of the mechanism of human platelet-rich plasma in regulating and controlling human epidermal stem cells for promoting wound re-epithelialization at transcriptome level

Xu Pengcheng, He Weifeng, Cheng Biao

2020, 36(10): 915-922. doi: 10.3760/cma.j.cn501120-20200707-00341

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Objective To analyze target genes of human platelet-rich plasma (PRP) in regulating and controlling human epidermal stem cells (ESCs). Methods (1) The discarded foreskin tissues were collected from 6 male patients of the First Affiliated Hospital of Army Medical University after urological surgery. The patients aged 5 to 25 years with good health and without urinary system infection. Human ESCs were cultivated using quick attachment method, and were subjected to morphological observation and identification. Venous blood sample in the volume of 40 mL was collected from a female healthy volunteer (aged 29 years) of General Hospital of Southern Theater Command of PLA, and PRP was extracted by second centrifugation method. (2) The successfully cultured primary human ESCs were divided into control group and PRP group according to the random number table, with 3 wells in each group. The cells in control group were not specially treated. In PRP group, PRP was added to the ESC medium to achieve final volume fraction of 2.5% after the cells were adhered for 12 hours. RNA was extracted, and transcriptome sequencing and data analysis of human ESCs of two groups were performed using RNA sequencing technology. Using the false discovery rate less than 0.05 and the fold change more than or equal to 4 as the standard, the differentially expressed genes were screened by Dr. Tom data mining system. Gene ontology (GO) enrichment analysis was performed on the obtained differentially expressed genes to find out the GO entries with significant enrichment. Then Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation analysis was used to further analyze the biological processes or metabolic pathways in which differentially expressed genes might be involved. Finally, the genes related to re-epithelialization and significantly differentially expressed were selected, and the differential expression of genes was verified by real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR). Data were statistically analyzed with independent-samples t test. Results (1) The cultured cells were cloned with a paving stone-like shape and positive rate of CD49f of 95.132% and CD71 of 0.006%, which proved that the primary culture of ESCs was successful. (2) The quality control data analysis showed that the selected samples had better quality and higher sequence alignment rate, which met the requirements of sequencing. (3) Sequencing data showed that there were a total of 449 differentially expressed genes between the two groups, including 354 up-regulated genes and 95 down-regulated genes. Further cluster analysis determined that there were 18 significantly up-regulated genes and 5 significantly down-regulated genes between the two groups. GO enrichment analysis and KEGG pathway annotation analysis showed that the significantly differentially expressed genes were mainly enriched in the epidermis construction and keratinization process, which also might be related to interleukin 17 signaling pathway. (4) Keratin 19, keratin 10, and S100A7 genes which were related to the process of re-epithelialization and significantly differentially expressed were selected for verification. Real-time fluorescent quantitative RT-PCR showed that compared with those of control group, the mRNA expressions of keratin 19 and S100A7 of cells in PRP group were significantly increased (t=10.270, 5.690, P<0.01), while the mRNA expression of keratin 10 was significantly decreased (t=7.306, P<0.01), which was consistent with the result of sequencing data. Conclusions PRP regulates function of human ESCs and promotes wound re-epithelialization involving transcriptional regulation of multiple genes, including keratin 19, keratin 10, and S100A7. In-depth exploration of the possible regulatory network of PRP affecting human ESCs will provide the basis for its subsequent clinical application.

Study on mechanisms of interleukin-17A regulating the expressions of interleukin-1β and interleukin-23 in mouse keratinocytes

Li Yashu, Zhang Xiaorong, Yu Meijie, Hu Xiaohong, Yang Jiacai, Huang Yong, Luo Gaoxing, He Weifeng

2020, 36(10): 923-929. doi: 10.3760/cma.j.cn501120-20200619-00315

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Objective To investigate the mechanisms of interleukin-17A (IL-17A) regulating the expressions of IL-1β and IL-23 in mouse keratinocytes (KCs). Methods Primary KCs were isolated from the skin of 400 newborn male and female wild type C57BL/6 mice and cultured in 24-well plates with Roswell Park Memorial Institute 1640 medium containing fetal bovine serum in the volume fraction of 10% for the following experiments. (1) The cells were divided into phosphate buffer solution (PBS) control group and IL-17A stimulation group according to the random number table (the same grouping method below), which were cultured with 10 μL PBS or 10 μL IL-17A in the mass concentration of 100 ng/mL for 6 hours, respectively. The expression levels of IL-1β and IL-23 mRNA in cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR), with 3 samples in each group. (2) The cells were divided into dimethyl sulfoxide (DMSO) control group, IL-17A+ DMSO group, IL-17A+ nuclear factor κB (NF-κB) inhibitor group, IL-17A+ signal transduction and activator of transcription 3 (STAT3) inhibitor group, IL-17A+ extracellular signal-regulated kinase 1 (ERK1) inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ c-Jun N-terminal kinase (JNK) inhibitor group. The reagents were added to cells in corresponding groups respectively and cultured for 6 hours. The volume of each reagent was 10 μL, the mass concentration of IL-17A was 100 ng/mL, and the molarity concentrations of NF-κB, STAT3, ERK1, ERK2, JNK signal pathway inhibitors PDTC, S3I-201, SCH772984, SCH772984, SP600125 were 5 μmol/L, 100 μmol/L, 4 nmol/L, 1 nmol/L, and 10 μmol/L, respectively. The expression levels of IL-1β mRNA and IL-23 mRNA in cells were detected by real-time fluorescence quantitative RT-PCR, with 3 samples in each group. (3) The cells were grouped and treated the same as those in experiment (1). The levels of NF-κB phosphorylation, STAT3 phosphorylation, ERK phosphorylation, and JNK phosphorylation were detected by Western blotting, with 3 samples in each group. Data were statistically analyzed with two-tailed Student t test, one-way analysis of variance, t test, and Bonferroni correction. Results (1) After culture of 6 hours, compared with those in PBS control group, the expression levels of IL-1β and IL-23 mRNA in cells in IL-17A stimulation group were significantly increased (t=13.46, 6.72, P<0.01). (2) After culture of 6 hours, the expression levels of IL-1β and IL-23 mRNA in cells in DMSO control group, IL-17A+ DMSO group, IL-17A+ NF-κB inhibitor group, IL-17A+ STAT3 inhibitor group, IL-17A+ ERK1 inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ JNK inhibitor group were 1.00±0.11, 4.01±0.32, 0.32±0.06, 1.76±0.43, 3.62±0.24, 3.80±0.43, 4.26±0.74 and 1.03±0.29, 4.08±0.34, 4.76±0.38, 4.70±0.21, 1.06±0.42, 0.92±0.21, 0.39±0.05, respectively. Compared with those in DMSO control group, the expression levels of IL-1β and IL-23 mRNA in cells in IL-17A+ DMSO group were significantly increased (t=9.24, 12.60, P<0.01). Compared with that in IL-17A+ DMSO group, the expression level of IL-1β mRNA was significantly decreased in cells in IL-17A+ NF-κB inhibitor group and IL-17A+ STAT3 inhibitor group (t=11.34, 6.91, P<0.01). Compared with that in IL-17A+ DMSO group, the expression level of IL-23 mRNA was significantly decreased in cells in IL-17A+ ERK1 inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ JNK inhibitor group (t=12.44, 13.03, 15.21, P<0.01). (3) After culture of 6 hours, compared with those in PBS control group, the levels of NF-κB phosphorylation, STAT3 phosphorylation, ERK phosphorylation, and JNK phosphorylation in cells in IL-17A stimulation group were significantly increased. Conclusions IL-17A promotes the transcription of IL-1β in mouse KCs through the phosphorylation of NF-κB and STAT3 pathways and IL-23 through the phosphorylation of ERK and JNK pathways.

Preparation and preliminary research on the characteristics of modified nano-bioglass hydrogel

Wei Lichun, Zhang Yijie, Huang Sha, Yao Bin, Li Xiang, Chen Xuyuan, Li Yan, Fu Xiaobing, Wu Xu

2020, 36(10): 930-938. doi: 10.3760/cma.j.cn501120-20190806-00337

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Objective To explore the preparation and preliminary research on the characteristics of modified nano-bioglass hydrogel. Methods (1) The nano-bioglass suspension was prepared by adding 67 mL nano-silica suspension into 400 mL saturated calcium hydroxide solution, and its suspension stability was observed. (2) The hydrogel with final mass fraction of 10% gelatin and 1% sodium alginate was prepared and set as control group. On the basis of the hydrogel in control group, the nano-bioglass suspension prepared in experiment (1) was added to prepare the hydrogel with the final mass fraction of 0.5% bioglass, 10% gelatin, and 1% sodium alginate, and the hydrogel was set as the experimental group. The gelling time at 4 and 25 ℃and the dissolution time at 37 ℃ of hydrogel in 2 groups were recorded, and the gelation at 4 and 25 ℃and dissolution condition at 37 ℃of the hydrogel in 2 groups were observed. The hydrogel in 2 groups were collected and cross-linked with 25 g/L calcium chloride solution after cold bath at 4 ℃, and the compression modulus was measured by Young′s modulus tester. In addition, the hydrogel in 2 groups were collected and cross-linked as before, and freeze-drying hydrogel was made at -20 ℃. The relative volumes were measured and the porosity of hydrogel in 2 groups was calculated. The number of sample in the experiment was 3. (3) Fibroblasts (Fbs) were isolated and cultured from 12 C57BL/6J mice of 24 hours old and the morphology was observed by inverted microscope, and the third passage of Fbs were cultured for the following experiment. Fbs were collected to make single cell suspension with the cell concentration of 1×10⁵/mL. The single cell suspension was divided into experimental group and control group according the random number table (the same grouping method below), which were added with hydrogel in experimental group and control group prepared in experiment (2), respectively. At culture hour 12, 24, and 48, cells of 3 wells in each group were collected to detect the survival rate by cell counting kit 8 method. (4) The third passage Fbs were collected to prepare the single cell suspension with the cell concentration of (3.0~4.5)×10⁷/mL, which was divided into experimental group and control group, with 1 tube in each group. The single cell suspension in 2 groups were added with green fluorescent probe DIO for staining and then added with 9 mL hydrogel in experimental group and control group prepared in experiment (2), respectively. The mixed solution of Fbs and hydrogel in 2 groups was cross-linked as before to make cell-loaded hydrogel. On culture day 3, the survival of cells in the hydrogel was observed by laser confocal microscope. The cell-loaded hydrogel was prepared as before and without added with green fluorescent probe DIO. On culture day 7, the adhesion and extension of cells in the hydrogel were observed by scanning electron microscope. (5) Twelve 6-week-old female BALB/c-nu nude mice were collected and divided into experimental group and control group, with 6 mice in each group. A round full-thickness skin defect wound with diameter of 1 cm was made on the back of each mouse. Immediately after injury, one cell-loaded hydrogel block in the experimental group and the control group prepared in experiment (4) was placed in the wound of each mouse in the experimental group and the control group, respectively. On post injury day (PID) 7 and 14, 3 nude mice in each group were sacrificed to collect the wound and wound margin tissue, which was stained with hematoxylin-eosin to observe the wound healing. Data were statistically analyzed with independent sample t test. Results (1) The nano-bioglass particles could be uniformly dispersed in water and had good suspension stability. (2) The hydrogels of the 2 groups were molten at 37 ℃, and no precipitation of particle was observed. The dissolving time of the hydrogel in the experimental group and the control group at 37 ℃ was 5 and 10 min, respectively. The gelation time of the hydrogel in the experimental group and the control group at 25 ℃ was 30 and 180 min, respectively, and the gelation time of the 2 groups at 4 ℃ was 5 and 10 min, respectively. The compression modulus of hydrogel in the experimental group was (53±6) kPa, which was significantly higher than (23±6) kPa in the control group (t=6.364, P<0.01). The porosity of the hydrogel in the experimental group was (86.1±2.1)%, which was similar to (88.2±4.4)% in the control group (t=1.210, P>0.05). (3) The cells were in long fusiform, and the proportion of nuclei was high, which was accorded with the morphological characteristics of Fbs. At culture hour 12, 24, and 48, the survival rate of cells in the experimental group was (84±4)%, (89±4)%, and (130±10)%, which was similar to (89±5)%, (90±4)%, and (130±11)% in the control group, respectively (t=1. 534, 0.611, 0.148, P>0.05). (4) On culture day 3, the cells in the two groups had complete morphology in the hydrogel, no nuclear lysis or disappearance were observed, the cytoplasm remained intact, and the fluorescence intensity of the cells in the experimental group was significantly stronger than that in the control group. On culture day 7, the cells in the experimental group and the control group adhered and stretched in the hydrogel, and the number of cells in the experimental group adhered to the hydrogel was significantly more than that in the control group. On PID 7, the wound area of the nude mice in the control group and the experimental group were reduced, the reduction area of mice in the experimental group was more obvious, and a large amount of inflammatory cells were seen in and around the wound in the 2 groups. On PID 14, the wound area of the nude mice in the control group was larger than that of the experimental group, and the number of inflammatory cells in and around the wound was significantly more than that in the experimental group. Conclusions Nano-bioglass hydrogel possesses good physical, chemical, and biological properties, cell loading potential, and the ability to promote wound healing, which means it has a good potential in clinical application.

Effect of fluid resuscitation guided by pulse contour cardiac output monitoring technology on organ function in extremely severe burn patients

Jiang Nanhong, Wang Deyun, Li Feng, Xi Maomao, Xie Weiguo

2020, 36(10): 939-946. doi: 10.3760/cma.j.cn501120-20190811-00345

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Objective To investigate the effect of fluid resuscitation guided by pulse contour cardiac output (PiCCO) monitoring technology on the organ function in extremely severe burn patients. Methods From May 2015 to March 2019, 52 patients with extremely severe burn hospitalized in Tongren Hospital of Wuhan University & Wuhan Third Hospital, meeting the inclusion criteria, were recruited to conduct a prospectively randomized control study. The patients were divided into PiCCO monitoring rehydration group (25 cases, 17 males and 8 females) and traditional rehydration group (27 cases, 20 males and 7 females) according to the random number table, with the ages of (47±9) and (49±8) years respectively. After admission, all the patients were rehydrated according to the rehydration formula of the Third Military Medical University during shock stage. In traditional rehydration group, fluid resuscitation of the patients was performed by monitoring the traditional shock indicators such as urine volume and central venous pressure, while PiCCO monitoring was performed in patients in PiCCO monitoring rehydration group, and the global end-diastolic volume index combined with the other relevant indicators of PiCCO monitoring were used to guide rehydration on the basis of the monitoring indicators of traditional rehydration group. The rehydration coefficients and urine volumes per kilogram of body weight per hour during the first and second 24 h post injury were compared between the two groups, which were compared with the corresponding rehydration scheme value of the Third Military Medical University (hereinafter referred to as the scheme value) at the same time. The total rehydration volumes within post injury hour (PIH) 8 and during the first and second 24 h post injury, the urine volumes per hour during the first and second 24 h post injury, and the levels of creatinine, urea nitrogen, lactate clearance rate, procalcitonin, creatine kinase isoenzyme (CK-MB) in blood and mean arterial pressure (MAP) on post injury day (PID) 1, 2, and 3 were measured. The incidence of complications, the application case number of mechanical ventilation, and the mechanical ventilation time within PID 28 were analyzed. Data were statistically analyzed with analysis of variance for repeated measurement, t test, Bonferroni correction, Mann-Whitney U test, chi-square test, and Fisher′s exact probability method test. Results During the second 24 h post injury, the rehydration coefficient of patients in traditional rehydration group was significantly higher than the scheme value (t=5.120, P<0.01). During the first and second 24 h post injury, the rehydration coefficients of patients in PiCCO monitoring rehydration group were significantly higher than the scheme values (t=3.655, 10.894, P<0.01) and those in traditional rehydration group (t=3.172, 2.363, P<0.05 or P<0.01). Within PIH 8, the total rehydration volumes of patients between the two groups were similar. During the first and second 24 h post injury, the total rehydration volumes of patients in PiCCO monitoring rehydration group were significantly higher than those in traditional rehydration group (t=4.428, 3.665, P<0.01). During the first and second 24 h post injury, the urine volumes per kilogram of body weight per hour of patients in traditional rehydration group were significantly higher than the schema values (t=4.293, 6.362, P<0.01), and the urine volumes per kilogram body weight per hour of patients in PiCCO monitoring rehydration group were significantly higher than the schema values (t=6.461, 8.234, P<0.01). The urine volumes per kilogram of body weight per hour and urine volumes per hour of patients in PiCCO monitoring rehydration group during the second 24 h post injury were significantly higher than those in traditional rehydration group (t=2.849, 3.644, P<0.05 or P<0.01). The creatinine levels of patients between the two groups on PID 1, 2, and 3 were similar. The urea nitrogen levels of patients in PiCCO monitoring rehydration group on PID 1, 2, and 3 were (6.8±1.5), (5.6±1.4), (4.4±1.4) mmol/L respectively, which were significantly lower than (8.6±1.8), (6.6±1.5), (5.5±1.4) mmol/L in traditional rehydration group (t=3.817, 2.511, 2.903, P<0.05 or P<0.01). The lactate clearance rates of patients in PiCCO monitoring rehydration group on PID 1, 2, and 3 were significantly higher than those in traditional rehydration group (t=2.516, 4.540, 3.130, P<0.05 or P<0.01). The procalcitonin levels of patients in PiCCO monitoring rehydration group on PID 2 and 3 were significantly lower than those in traditional rehydration group (Z=-2.491, -2.903, P<0.05). The CK-MB level of patients in PiCCO monitoring rehydration group on PID 3 was (35±10) U/L, which was significantly lower than (51±16) U/L in traditional rehydration group (t=4.556, P<0.01). The MAP levels of patients between the two groups on PID 1, 2, and 3 were similar. Within PID 28, the incidence of complications of patients in traditional rehydration group was significantly higher than that in PiCCO monitoring rehydration group (χ²=4.995, P<0.05), and the application case number of mechanical ventilation and the mechanical ventilation time of patients between the two groups were similar. Conclusions The use of PiCCO monitoring technology to guide the early fluid resuscitation of extremely severe burn patients is beneficial for accurate determination of the fluid volume required by the patients and reduction of organ injury caused by improper rehydration.

Recurrence and influencing factors of diabetic foot ulcer in patients with type 2 diabetes mellitus

Shen Jinfu, Jiang Ruimei, Wang Zhuoqun, Li Mao, Li Juan, Xie Shuyong, Kang Jingjing

2020, 36(10): 947-952. doi: 10.3760/cma.j.cn501120-20190726-00315

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Objective To investigate the recurrence and influencing factors of diabetic foot ulcer in patients with type 2 diabetes mellitus. Methods Totally 185 type 2 diabetes patients with new-onset of diabetic foot ulcers admitted to Fuyang People′s Hospital of Anhui Province from January 2011 to December 2015 were enrolled in this study, including 120 males and 65 females, aged 40-79 years. All the patients were followed up for 3 years, and their clinical data were retrospectively analyzed by the case-control study. The Kaplan-Meier cumulative recurrence curve was drawn according to the 3-year cumulative recurrence rate of diabetic foot ulcers. The time to visit, toe involvement, and amputation of involved toes in patients with recurrent diabetic foot ulcer were counted at the initial onset and the recurrence of the ulcers, respectively, and the data were statistically analyzed with t test and chi-square test. According to the recurrence of diabetic foot ulcers, the patients were divided into foot ulcer recurrence group and foot ulcer non-recurrence group. The gender, age, course of diabetes mellitus, length of hospital stay, visit time, body mass index, glycosylated hemoglobin HbA1c, total bilirubin, albumin, creatinine, cholesterol, low density lipoprotein (LDL), high density lipoprotein (HDL), triglycerides, hemoglobin, white blood cell count, toe involvement, toe amputation, ankle-brachial index, diabetic retinopathy (DR), diabetic peripheral neuropathy (DPN), diabetic nephropathy (DN), history of hypertension, cardio-cerebrovascular disease, smoking, residence, solitary life, and walking disorder of patients between the two groups were compared, and the data were statistically analyzed with t test and chi-square test. Log-rank test was performed on the indexes with P<0.1 in comparison between two groups, and the indexes with statistically significant differences in Log-rank test were analyzed by multivariate Cox regression analysis to screen the influencing factors of recurrence of diabetic foot ulcer. Results (1) The 3-year cumulative recurrence rate of diabetic foot ulcers in 185 patients with type 2 diabetes mellitus was 47.0% (87/185). (2) For 87 patients with diabetic foot ulcer recurrence, compared with that at the initial onset of the ulcers, the visit time was significantly shorter (t=10.593, P<0.01), the toe amputation rate was significantly increased (χ²=5.118, P<0.05), but there was no obvious change in toe involvement at the recurrence of the ulcers. (3) There were statistically significant differences in age, course of diabetes mellitus, length of hospital stay, body mass index, glycosylated hemoglobin HbA1c, total bilirubin, albumin, creatinine, cholesterol, LDL, HDL, hemoglobin, white blood cell count, gender, toe amputation, ankle-brachial index, DR, history of cardio-cerebrovascular disease, solitary life, and walking disorder of patients between foot ulcer recurrence group (87 patients) and foot ulcer non-recurrence group (98 patients) (t=5.123, 4.242, 5.324, -24.572, 6.102, -1.984, -9.747, 3.226, 3.076, 3.646, -4.683, -7.502, 8.095, χ²=5.621, 18.433, 4.546, 5.785, 9.655, 7.625, 7.886, P<0.05 or P<0.01), while the rest of the indexes of patients between the two groups were similar. Log-rank test showed that the two groups had statistically significant differences in age, course of diabetes mellitus, length of hospital stay, glycosylated hemoglobin HbA1c, total bilirubin, albumin, creatinine, ankle-brachial index, DPN, and walking disorder (χ²=210.046, 44.837, 34.107, 98.685, 66.532, 294.451, 260.554, 5.012, 6.818, 11.160, P<0.05 or P<0.01). Age, total bilirubin, albumin, DPN, and walking disorder were the influencing factors for the recurrence of diabetic foot ulcers in patients with type 2 diabetes mellitus (hazard ratio=1.024, 0.678, 0.849, 2.335, 4.099, 95% confidence interval=1.001-1.047, 0.558-0.823, 0.797-0.904, 1.280-4.258, 2.044-8.223, P<0.05 or P<0.01). Conclusions The 3-year cumulative recurrence rate of diabetic foot ulcers in patients with type 2 diabetes mellitus is relatively high, with the influencing factors being age, total bilirubin, albumin, DPN, and walking disorder.

Effects of smoking on the wound healing of stage 4 pressure ulcers in rats

Song Meiyi, Li Xian, Liu Shanshan, Wang Yao, Zhao Zhihong, Wang Yue, Chen Ziye

2020, 36(10): 953-958. doi: 10.3760/cma.j.cn501120-20190827-00361

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Objective To explore the effect of smoking on the wound healing of stage 4 pressure ulcers in rats. Methods Fifty male Sprague-Dawley rats aged 6-8 weeks were divided into simple pressure ulcer group and smoking+ pressure ulcer group according to the random number table, with 25 rats in each group. After the rats in the smoking+ pressure ulcer group received passive smoking intervention for 12 weeks, an iron plate was placed in the back muscle of each rat in 2 groups, and a magnet was placed outside the skin at the corresponding position of the iron plate for 2 h at each time, with 5 times a day and continuously for 6 days to reproduce stage 4 pressure ulcer model. Immediately after establishing the model, 3 rats in each group were sacrificed and wound tissue was collected, and hematoxylin-eosin staining was applied to observe the pathological changes of the wounds. On 1, 3, 7, and 14 day (s) after establishing the model, 3 rats in each group were collected to measure the pressure ulcer wound area by the paper jam method. After measurement of the wound area, the rats were sacrificed and the wound tissue was collected, and the protein expression levels of matrix metalloproteinases 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in wound tissue were detected by immunohistochemical method, and the ratio of MMP-9/TIMP-1 was calculated.The wound healing time of the remaining 10 rats in each group was recorded. Data were statistically analyzed with analysis of variance for factorial design, two independent sample t test, and Bonferroni correction. Results (1) Immediately after establishing the model, muscle fiber necrosis and dissolution with large areas were seen on the wound, the myofibrils arranged loosely, and more lymphocytes and monocytes infiltration were seen around the wound of rats in simple pressure ulcer group. A large number of necrotic myofibers were dissolved and gradually disappeared, the myofibrils arranged loosely, and the number of diffuse lymphocytes and monocyte infiltration in wound of rats in smoking+ pressure ulcer group were significantly higher than those in simple pressure ulcer group. (2) The wound areas of rats in smoking+ pressure ulcer group were significantly larger than those in simple pressure ulcer group on 1, 3, 7, and 14 day (s) after establishing the model (t=3.019, 2.549, 2.181, 3.674, P<0.05 or P<0.01). (3) On 1 to 14 days after establishing the model, the protein expression levels of MMP-9 and TIMP-1 in the wound tissue and the ratio of MMP-9/TIMP-1 of rats in the two groups increased first and then decreased. On 1, 3, 7, and 14 day (s) after establishing the model, the protein expression levels of MMP-9 in the wound tissue and the ratio of MMP-9/TIMP-1 of rats in smoking+ pressure ulcer group were significantly higher than those in simple pressure ulcer group (t=4.783, 4.508, 6.325, 7.204, 3.078, 2.989, 4.081, 4.696, P<0.05 or P<0.01), and the protein expression levels of TIMP-1 in wound tissue of rats in the two groups were similar. (4) The wound healing time of rats in smoking+ pressure ulcer group was (48.9±2.6) d, which was significantly longer than (35.2±2.3) d of simple pressure ulcer group (t=12.477, P<0.05). Conclusions Smoking can up-regulate the expression of MMP-9 in pressure ulcer wound and result in an imbalance of MMP-9/TIMP-1, thereby affecting the wound healing of stage 4 pressure ulcers in rats.

Systematic review of the qualitative researches on care experience of caregivers of burn children

Liu Jiahui, Xiong Yu, Hu Zhaoyang, Jiang Daofeng

2020, 36(10): 959-965. doi: 10.3760/cma.j.cn501120-20200108-00013

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Objective To systematically review the care experience of caregivers of burn children, so as to provide references for guiding the continuing care in hospitals, communities, and homes. Methods Databases including Cochrane Library, PubMed, ScienceDirect, ProQuest, Web of Science, and CINAHL were retrieved with the search terms of " burn" , " care/caregivers/nursing/father/mother/relatives" , " needs/perceptions/exceptions/attitudes/feelings/demands/experiences" , " qualitative research" , and the Chinese Journals Full-text Database, China Biology Medicine disc, VIP Database, and Wanfang Data were retrieved with the search terms in Chinese version of "烧伤/烧伤患儿/烧伤幼儿" , "照顾者/照护者/照料者/照护/父亲/母亲/家属" , "需求/认知/期望/态度/感受/体验" , "质性研究" to search the qualitative researches on care experience of caregivers of burn children published from the establishment of the databases to November 2019. After screening and extracting the data, the quality evaluation criteria for qualitative research of the Australian Joanna Briggs Institute Evidence-Based Health Care Center and its integrative/aggregative synthesis method were used to assess the quality of the included literature and meta-integrate the research results respectively. Results A total of 16 studies and 269 caregivers were enrolled. The quality of one included literature was grade A, and the quality of 15 included literature was grade B. A total of 65 research results were extracted with totally 6 categories formed after summarization, and 2 integrated results obtained as follows: (1) The caregivers experienced heavy psychological pressure and burden in the care process, which had a significant impact on family, social relations, and daily life. (2) With the care time lapsing, through the support of all sectors of society and self-adjustment, the caregivers gradually accepted the reality and actively took various countermeasures, but they still faced many challenges in disease care. Conclusions The caregivers of burn children have many physical and mental health problems and face many care challenges. The government, medical and health institutions, and society should give a great attention to these issues, improve the social support system and security system, reduce the family-related pressure of burn children′s families, and improve the quality of family life.

Clinical characteristics of treadmill abrasion in children

Li Min, Zeng Xiangchun, Xie Weiguo

2020, 36(10): 966-968. doi: 10.3760/cma.j.cn501120-20191022-00408

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Objective To analyze the clinical characteristics of treadmill abrasion in children. Methods The medical records of 12 children with treadmill abrasion who were admitted to Tongren Hospital of Wuhan University & Wuhan Third Hospital (hereinafter referred to as the author′s unit) from August 2017 to August 2019 were retrospectively analyzed. The data of gender, age, time of seeing a doctor after injury, injury environment and cause, abrasion site, abrasion area, abrasion depth, and wound treatment method and outcome of children were analyzed. Results (1) The children included 5 males and 7 females, with the age of 2 to 6 years. Three children were treated on the same day after injury, and 9 children were treated 7 to 15 days after injury. (2) All the children were injured at home, and the abrasion was mainly caused by touching the running treadmill when their parents were using it. The contact area was mainly the gap between the rear track axle and the metal plate at the lower edge. (3) All the children had abrasion in the hands. The index finger, middle finger, and ring finger were the main involved fingers. The area of the abrasion was 2 cm×2 cm to 8 cm×5 cm, all of them were deep-partial thickness and above. (4) The children were mainly treated in outpatient department (9 children), and 10 children were conducted with conservative treatment. Eleven children had local scar formation, after anti-scar treatment and functional exercise, they recovered well without obvious dysfunction. Conclusions Hands in those children are the most vulnerable part of treadmill abrasion injury, with the index finger, middle finger, and ring finger being frequently involved. The wound area is generally small, and most of the wounds are deep abrasion injuries.

Early debridement combined with antidote in the treatment of one patient with chromic acid burn and acute kidney injury

Zhang Ying, Xu Gang, Luo Yi, Jin Junjun

2020, 36(10): 969-970. doi: 10.3760/cma.j.cn501120-20190813-00349

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Research status and prospect on autologous tissue-engineered skin as permanent graft

Kang Kun, Shao Yang, Song Guodong

2020, 36(10): 971-974. doi: 10.3760/cma.j.cn501120-20200204-00043

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