Objective To investigate the application of pulse contour cardiac output (PiCCO) monitoring technology in fluid resuscitation of severe burn patients in shock period. Methods From January 2015 to December 2019, 33 patients with severe burns who were hospitalized in Guangzhou Red Cross Hospital, meeting the inclusion criteria, were recruited into a retrospective cohort study with their clinical information collected. The patients were divided into PiCCO monitoring group with 15 cases (13 males and 2 females, aged (43±13) years) and routine monitoring group with 18 cases (14 males and 4 females, aged (39±9) years) according to the monitoring method used. After admission, all the patients were rehydrated following the rehydration formula of the Third Military Medical University for shock period. In routine monitoring group, the fluid resuscitation of patients was performed by monitoring indicators such as urine volume and blood pressure, while PiCCO monitoring was performed among patients in PiCCO monitoring group, and their fluid resuscitation was guided by the patient′s condition and the hemodynamic parameters (without pursuing normal levels of the parameters) of PiCCO monitoring on the basis of normal monitoring indicators in routine monitoring group. The colloids coefficients, the electrolyte coefficients (compared with the corresponding rehydration formula value of 0.75 mL·kg^-1·% total body surface area (TBSA)^-1 of the Third Military Medical University for shock period during the first 24 h post injury), the total rehydration coefficients, and the urine volumes during the first and second 24 h post injury, the lactic acid level, the base excess level, and the oxygenation index at admission and 24, 48 h after admission, and the mechanical ventilation time, the wound healing time, and the death ratio of patients in the two groups were recorded. The cardiac index, the global end-diastolic volume index (GEDVI), the intrathoracic blood volume index (ITBVI), the extravascular lung water index (EVLWI), and the systemic vascular resistance index (SVRI) of patients in PiCCO monitoring group at post injury hour 24, 48, and 72 and the abnormal cases were recorded. Data were statistically analyzed with Fisher′s exact probability test, independent-sample or one-sample

test, analysis of variance for repeated measurement, and Bonferroni correction. Results During the first 24 h post injury, the colloids coefficients of patients in PiCCO monitoring group was (0.69±0.15) mL·kg^-1·%TBSA^-1, which was significantly less than (0.85±0.16) mL·kg^-1·%TBSA^-1 in routine monitoring group (

=-2.612,

<0.05). Compared with the rehydration formula value of the Third Military Medical University for shock period, only the colloids coefficient of patients in routine monitoring group during the first 24 h post injury was significantly increased (

=2.847,

<0.05). There were no statistically significant differences between the two groups in the colloids coefficients of patients during the second 24 h post injury, or the electrolyte coefficients, the total rehydration coefficients, the urine volumes of patients during the first and the second 24 h post injury (

=0.579, -0.011, 0.417, -1.321, -0.137, 0.031, 1.348,

>0.05). The lactic acid level, the base excess level, the oxygenation index of patients at admission and 48 h after admission, and the oxygenation index of patients at 24 h after admission between the two groups were similar (

=-1.837, 0.620, 0.292, -1.792, 1.912, -0.167, 1.695,

>0.05). The levels of lactic acid and base excess of patients in PiCCO monitoring group were (4.8±1.4) and (1.2±5.5)mmol/L, respectively, which were significantly better than (7.0±1.5) and (-2.8±3.0) mmol/L in routine monitoring group at 24 h after admission (

=-3.904, 2.562,

<0.05 or

<0.01). There were no statistically significant differences between the two groups in the mechanical ventilation time or the wound healing time of patients (

=-0.699, -0.697,

>0.05), or the death ratio of patients (

>0.05). In PiCCO monitoring group, the GEDVI, and the ITBVI of patients were lower than the normal low values at post injury hour 24 and 48, which were in the normal range at post injury hour 72; the cardiac index of patients increased gradually and recovered to normal at post injury hour 48; the SVRI of patients increased significantly at post injury hour 24 and then gradually decreased to normal; the EVLWI average of patients at all time points post injury were less than 10 mL/kg. At post injury hour 24, most of the hemodynamic parameters of more than or equal to 8/15 patients in PiCCO monitoring group were abnormal, and the abnormal proportion decreased later. Conclusions On the basis of traditional monitoring indicators, the use of PiCCO monitoring technology combined with the patient′s condition (without pursuing normal levels of the parameters) in guiding the fluid resuscitation in severe burn patients can reduce the usage of colloid and better improve tissue perfusion, with the resuscitation effect being better than conventional monitoring.

Objective To explore the value of renal injury marker protein in early diagnosis of acute kidney injury (AKI) in burn patients with delayed resuscitation. Methods The retrospective case-control research was conducted. Forty-three burn patients with delayed resuscitation (27 males and 16 females, with age of 18-75 (35±3) years)who were admitted to Zhengzhou First People′s Hospital from May 2018 to May 2020 met the inclusion criteria. The patients were divided into AKI group with 23 patients and non-AKI group with 20 patients according to whether AKI occurred within 7 days after burns. The gender, age, deep partial-thickness burn area, full-thickness burn area, and acute physiology and chronic health evaluation Ⅱ of patients were compared between the two groups.The fluid supplement volume and serum creatinine at 12, 24, and 48 h after burn, serum albumin/fibrinogen ratio (AFR), urinary heat shock protein 70 (HSP70), tissue inhibitor of metalloproteinase-2 (TIMP-2)×insulin-like growth factor binding protein 7 (IGFBP-7), and neutrophil gelatinase associated lipocalin (NGAL)at 12, 24, 48, 72, 120, and 168 h after burn were detected.Data were statistically analyzed with Mann-Whitney

test, analysis of variance for repeated measurement, independent-samples

test, chi-square test and Bonferroni correction. The independent variable to predict the occurrence of AKI was screened by multi-factor logistic regression analysis. The receiver′s operating characteristic curve was drawn for predicting the occurrence of AKI in burn delayed resuscitation patients, and the area under the curve (AUC), the best threshold, and the sensitivity and specificity under the best threshold were calculated. Results The gender, age, deep partial-thickness burn area, full-thickness burn area, acute physiology and chronic health evaluation Ⅱ of patients in two groups were similar (

²=1.98,

=1.98, 1.99, 1.99, 1.99,

>0.05). The fluid supplement volume of patients in AKI group at 24 and 48 h after burn was significantly less than that in non-AKI group (

=15.37, 6.51,

<0.01). The serum creatinine of patients in AKI group at 12, 24, and 48 h after burn was significantly higher than that in non-AKI group (

=2.16, 5.62, 6.72,

<0.01). The serum AFR of patients in AKI group at 12, 24, 48, 72, 120, and 168 h after burn was significantly lower than that in non-AKI group (

=16.14, 35.35, 19.60, 20.47, 30.20, 20.17,

<0.01). The levels of urinary HSP70 of patients in AKI group at 12, 24, 48, 72, 120, and 168 h after burn were (6.89±0.87), (6.42±0.73), (5.81±0.72), (5.17±0.56), (4.63±0.51), (3.89±0.51) μg/L, which were significantly higher than (3.89±0.75), (3.57±0.63), (2.66±0.41), (1.83±0.35), (1.48±0.19), (1.28±0.19) μg/L in non-AKI group (

=12.00, 13.61, 17.39, 22.98, 26.34, 21.59,

<0.01). Urinary TIMP-2×IGFBP-7 and NGAL of patients in AKI group at 12, 24, 48, 72, 120, 168 h after burn were significantly higher than those in non-AKI group (

=26.94, 101.11, 35.50, 66.89, 17.34, 14.30, 14.00, 13.78, 12.32, 14.80, 21.36, 22.62,

<0.01). Urinary HSP70 and serum AFR at 12 h after burn, urinary TIMP-2×IGFBP-7 and NGAL at 24 h after burn were included into multi-factor logistic regression analysis (odds ratio=2.42, 3.47, 7.52, 5.61, 95% confidence interval=1.99-2.95, 1.86-3.92, 2.87-9.68, 2.14-14.69,

<0.01). For 43 patients with burn delayed resuscitation, the AUC of receiver′s operating characteristic curve of serum AFR at 12 h after burn for predicting AKI was 0.739 (95% confidence interval=0.576-0.903), the optimal threshold was 9.90, the sensitivity was 82%, and the specificity was 90%. The AUC of urinary HSP70 at 12 h after burn was 0.990 (95% confidence interval=0.920-1.000), the optimal threshold was 1.40 μg/L, the sensitivity was 98%, and the specificity was 96%. The AUC of urinary TIMP-2×IGFBP-7 at 24 h after burn was 0.715 (95% confidence interval=0.512-0.890), the optimal threshold was 114.20 μg²/L², the sensitivity was 91%, and the specificity was 95%. The AUC of urinary NGAL at 24 h after burn was 0.972 (95% confidence interval=0.860-1.000), the optimal threshold was 78 μg/L, the sensitivity was 95%, and the specificity was 96%. Conclusions Urinary HSP70 and NGAL have higher value in early diagnosis of AKI in burn patients with delayed resuscitation.

Objective To study the coagulation characteristics of adult patients with extensively severe burn in shock stage and its alarming value. Methods Retrospective cohort study was performed on medical records of 37 adult patients with extensively severe burn who were admitted to the First Affiliated Hospital of Naval Medical University from January 2014 to December 2019 and met the inclusion criteria. The patients were divided into survival group (

=23, 17 males and 6 females, aged 41 (31, 51) years) and death group (

=14, 11 males and 3 females, aged 50 (43, 58) years) according to the prognosis of within 60 d after burn. Basic data of patients in the two groups and their routine coagulation indexes during shock period including prothrombin time (PT), thrombin time, activated partial thromboplastin time (APTT), D-Dimer, fibrinogen degradation product (FDP), fibrinogen, platelet, and international normalized ratio (INR) were recorded. Data were statistically analyzed with Wilcoxon rank sum test and Fisher′s exact probability test, prognosis-related factors was analyzed with single factor and multivariate logistic regression analysis (α selected=0.05, α excluded=0.1), and receiver operating characteristic (ROC) curve analysis were established to screen out the risk factors. All the patients were grouped into high score group and low score group according to the optimal threshold value, Kaplan-Meier method was used for survival analysis and Log-rank test was performed between the two groups. Results Total burn surface area (TBSA) of patients in death group was obviously larger than that in survival group (

=2.980,

<0.01), while there were no statistically significant difference in the other indexes between the two groups (

>0.05). Compared with those in survival group (16.10 (14.30, 16.90) s, 40.80 (36.20, 42.80) s, 1.30 (1.10, 1.40)), PT (18.70 (16.30, 22.70) s), APTT (46.45 (41.00, 57.10) s) and INR (1.55 (1.30, 1.96)) of patients in death group were significantly increased (

=2.540, 2.330, 2.300,

<0.05), there were no statistically significant difference in the other indexes between the two groups (

>0.05). Single factor logistic regression analysis showed TBSA, PT, and APTT were factors related to death of adult patients with extensively severe burn within 60 d after burn (odds ratio (OR)=1.190, 1.214, 1.109, 95% confidence interval (CI)=1.053-1.346, 1.008-1.461, 1.012-1.215,

<0.05 or

<0.01). FDP and INR were potential factors related to death of adult patients with extensively severe burn within 60 d after burn (OR=1.040 and 4.559, 95% CI =0.998-1.083 and 0.918-22.641,

<0.1). Multivariate logistic stepwise regression was used to build models of APTT+ FDP+ TBSA and APTT+ FDP. Area under the curve (AUC) of APTT+ FDP+ TBSA model score was 0.944 (95% CI= 0.873-1.000), which was higher than AUC of APTT+ FDP model score (0.843, 95% CI=0.713-0.973) by ROC curve analysis. Optimal threshold value of APTT+ FDP+ TBSA model score was -0.879 4 with sensitivity of 100% (95% CI=100%-100%) and specificity of 87% (95% CI=74%-100%). Survival ratio of patients in high score group with optimal threshold value higher than -0.879 4 was significantly lower than that in low score group with optimal threshold value lower than -0.879 4,

²=27.090,

<0.01. Conclusions The coagulation state of adult patients with extensively severe burn in shock stage is characterized with procoagulant and hemostatic dysfunctions accompanied by enhanced fibrinolytic activity. The risk of death is significantly increased in adult patients with extensively severe burn with APTT+ FDP+ TBSA model score higher than -0.879 4.

Objective To observe the effect of immunofluorescence double staining for foamy macrophages and

(MTB) in paraffin-embedded tissue of clinical tuberculous wound, in comparison with three routine staining methods. Methods The experimental method was used. From April 2019 to May 2020, 10 patients with tuberculous wound (5 males and 5 females, aged 28-77 years) meeting the inclusion criteria were treated in the Department of Burns and Plastic & Wound Repair Surgery of Xiang′an Hospital of Xiamen University. The paraffin-embedded wound tissue were collected during extended debridement and preserved in the Department of Pathology of this hospital. Forty paraffin sections were made from the wound tissue of each patient. Hematoxylin-eosin (HE) staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining were performed respectively, with 10 sections in each method. The section rejection rate of four staining methods were calculated. The recognition and detection of wound granuloma tissue in the four staining methods were observed and counted, and the recognition and detection of foamy macrophages in the wound tissue stained with four methods were observed. The MTB detection in the wound granuloma tissue and non-granuloma tissue in the four staining methods were compared. The subtyping and distribution of foamy macrophages and detection rate of MTB in the wound granuloma tissue and non-granuloma tissue, the morphologic clarity of foamy macrophages, as well as the non-specific staining rate and the loss rate of positive reaction of MTB and foamy macrophages by Ziehl-Neelsen and immunohistochemical double staining were compared with those of immunofluorescence double staining. Data were statistically analyzed with Fisher′s exact probability test, one-way analysis of variance, independent sample

test and Wilcoxon signed rank test. Results The section rejection rate of HE staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, and immunofluorescence double staining were 3% (3/100), 1% (1/100), 6% (6/100), and 2% (2/100), respectively. There was no statistically significant difference among the four groups (

=0.26). All the four staining methods could identify granuloma tissue, and the number of granuloma structures was similar (

=1.284,

=0.28). All the four staining methods were able to identify foamy macrophages in the wound tissue, which was detected in each section. No MTB was observed in the wound granuloma tissue or non-granuloma tissue by HE staining or immunohistochemical staining. MTB was observed distributing in the wound granuloma tissue and non-granuloma tissue by Ziehl-Neelsen and immunohistochemical double staining and immunofluorescence double staining, and most MTB distributed in the wound granuloma tissue. Ziehl-Neelsen and immunohistochemical double staining could not distinguish foamy macrophages engulfed MTB from that non-engulfed MTB. Immunofluorescence double staining showed that foamy macrophages engulfed MTB mostly distributed in the wound granuloma tissue, and the foamy macrophages non-engulfed MTB mostly distributed in the wound non-granuloma tissue. The detection rates of MTB in wound granuloma and non-granuloma tissue in immunofluorescence double staining were (89.00±0.08)% and (82.67±0.05)%, respectively, which were significantly higher than (54.56±0.14)% and (44.44±0.13)% in Ziehl-Neelsen and immunohistochemical double staining (

=-12.495, -7.961,

<0.01). Compared with that of Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining showed better foamy macrophages clarity in wound tissue (

=-3.162,

<0.01). The nonspecific staining rate and positive reaction loss rate of MTB and foamy macrophages in wound tissue of immunofluorescence double staining were (9.11±0.07)% and (9.22±0.07)%, respectively, which were significantly lower than (20.67±0.06)% and (44.00±0.12)% of Ziehl-Neelsen and immunohistochemical double staining (

=4.569, 15.519,

<0.01). Conclusions Compared with HE staining, immunohistochemical staining, and Ziehl-Neelsen and immunohistochemical double staining, the immunofluorescence double staining is easy to operate, giving clear and intuitive images. It allows accurate imaging co-localization of MTB and foamy macrophages in paraffin-embedded tissue of clinical tuberculous wound.

Objective To investigate the effects and mechanism of sodium hydrosulfide on rat epidermal cells intervened with serum from burned rat (hereinafter referred to as burn serum). Methods The experimental research method was used. Ten male Sprague-Dawley (SD) rats aged eight months were taken to prepare normal rat serum (hereinafter referred to as normal serum), 30 male SD rats aged eight months were taken to prepare burn serum after full-thickness burn, and epidermal cells (the third passage)isolated from 10 SD rats born one day were used for the experiments. The cells were divided into normal serum group treated with normal serum and burn serum group treated with burn serum. Cell counting kit 8 method was used to detect cell survival rate after 1, 2, 4, 6, and 8 h of culture, respectively, to screen the subsequent intervention time of burn serum. The cells were divided into burn serum control group treated only with burn serum and 50, 100, 150, 200, 250 μmol/L sodium hydrosulfide groups treated with burn serum+ sodium hydrosulfide at corresponding final molarity. After 30 min of culture following the burn serum intervention, the cell survival rate was detected as above to screen the subsequent intervention concentration of sodium hydrosulfide. The cells were divided into burn serum control group treated with burn serum only and sodium hydrosulfide only group, glibenclamide only group, and sodium hydrosulfide+ glibenclamide group treated with burn serum+ corresponding reagents. After 5, 10, 15 min of culture following the burn serum intervention, the cell survival rate was detected as above to screen the subsequent intervention time of glibenclamide. The cells were divided into burn serum control group treated with burn serum and sodium hydrosulfide only group, glibenclamide only group, and sodium hydrosulfide+ glibenclamide group treated with burn serum+ corresponding reagents. After completing corresponding culture time of each reagent, the mitochondria were extracted to detect cytochrome c oxidase (CCO) activity using a spectrophotometer, and the protein expression level of adenosine triphosphate (ATP)-sensitive potassium channel was detected by Western blotting. Except for the number of samples for ATP-sensitive potassium channel protein detection, which was 3, the number of samples for the other indicators was 10. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, least significant difference (LSD)-

test, LSD test, and Bonferroni correction. Results Compared with that of normal serum group, the cell survival rate was significantly decreased in burn serum group after only 4 and 6 h of culture (

=4.02, 6.42,

<0.05). An overall comparison showed statistically significant differences in cell survival rate among the time points within normal serum group and burn serum group (

=19.74, 4.48,

<0.05 or

<0.01). Four hours of culture was selected as the subsequent intervention time of burn serum. After 30 min of culture following the burn serum intervention, compared with that of burn serum control group, only 150, 200, 250 μmol/L sodium hydrosulfide groups had a significantly higher cell survival rate (

<0.01), thus 150 μmol/L was selected as the subsequent intervention concentration of sodium hydrosulfide. Compared with that of burn serum control group, the cell survival rate decreased significantly in glibenclamide only group after 5 and 15 min of culture following burn serum intervention (

<0.05) and increased significantly in glibenclamide only group after 10 min of culture following the burn serum intervention and sodium hydrosulfide only group at each time point (

<0.05 or

<0.01). The cell survival rate in sodium hydrosulfide+ glibenclamide group was significantly lower than that of sodium hydrosulfide only group at each time point (

<0.05). The difference in cell survival rate was statistically significant among the time points within glibenclamide only group (

=11.81,

<0.01). Five minutes of culture was selected as the subsequent intervention time of glibenclamide. After 35 min of culture following the burn serum intervention, compared with (1.62±0.08) nmol·min^-1·mg^-1 and 0.682±0.063 in burn serum control group, the CCO activity of cells and the protein expression level of ATP-sensitive potassium channel were significantly increased in sodium hydrosulfide only group ((1.99±0.09) nmol·min^-1·mg^-1 and 0.932±0.014,

<0.01) and significantly decreased in glibenclamide only group ((1.44±0.09) nmol·min^-1·mg^-1 and 0.600±0.012,

<0.01); the CCO activity of cells and the protein expression level of ATP-sensitive potassium channel in sodium hydrosulfide+ glibenclamide group ((1.79±0.06) nmol·min^-1·mg^-1 and 0.744±0.071) was significantly lower than those of sodium hydrosulfide only group (

<0.05 or

<0.01). Conclusions Sodium hydrosulfide can improve the survival rate of rat epidermal cells after burn serum intervention, by a mechanism which is related to the alleviation of epidermal cell mitochondrial damage and mediated by ATP-sensitive potassium channel.

Objective To explore the effects of Freund′s complete adjuvant on autophagy protein expression in rat tuberculous wound model. Methods The experimental research method was used. In the first batch, twelve 6-week-old male Sprague-Dawley (SD) rats were sensitized by subcutaneous injection of Freund′s complete adjuvant into the hips. Three weeks later, the rats were infected with attenuated Bacille Calmette-Guérin (BCG) subcutaneously on both sides of the back spine. After establishing the tuberculosis wound rat model, according to the random number table (the same grouping method below), the rats were divided into 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group, with 3 rats in each group, with continuous normal feeding to the corresponding days after infection. In the second batch, twenty-three 6-week-old male SD rats were divided into blank control group (

=3, normal feeding without any treatment), BCG alone group (

=5), BCG+ rapamycin group (

=6), BCG+ 3-methyladenine group (

=6), and BCG+ starvation group (

=3). The last 4 groups of rats were sensitized as before, and infected as before 1 week later. Rats in BCG alone group were fed normally without any treatment. Rats in BCG+ rapamycin group or BCG+ 3-methyladenine group were intraperitoneally injected with rapamycin or 3-methyladenine once every other day and fed normally. Rats in BCG+ starvation group were fasted for 48 hours after infection and then fed normally. All the rats in the first batch of 4 groups were sacrificed on the corresponding days after infection, and the tissue where the buttocks were injected with Freund′s complete adjuvant was harvested; the tissue of rats in the second batch of BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were harvested the same as before 7 days after infection, and all the rats in blank control group were taken the same tissue at the same time point. Hematoxylin-eosin staining was performed to observe the structure and morphology of cells in the tissue harvested; immunohistochemistry was used to observe the protein expressions of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B) in the tissue harvested. Data were statistically analyzed with Kruskal-Wallis test and Bonferroni correction. Results Inflammatory cell infiltration was observed in the tissue of rats where the Freund′s complete adjuvant was injected in 8 d infection group, granuloma formation was seen in 15 d infection group, part of tissue cell necrosis was seen in 32 d infection group and 43 d infection group, and cell necrosis in 43 d infection group was worse than that in 32 d infection group. Seven days after infection, inflammatory cell infiltration was seen in the tissue of rats where the Freund′s complete adjuvant was injected in BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group, while regular arrangement of cells and no inflammatory cell infiltration were observed in blank control group. There were no statistically significant differences in the protein expressions of Beclin-1 or LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group (

=1.923, 5.821,

>0.05). Seven days after infection, the protein expressions of Beclin-1 and LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in blank control group, BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were respectively 0.325% (0.250%, 0.360%), 3.225% (1.340%, 3.987%), 4.823% (2.630%, 6.559%), 4.216% (1.790%, 5.969%), 1.765% (0.865%, 2.649%), and 0.301% (0.264%, 0.516%), 2.865% (1.455%, 5.768%), 1.033% (0.398%, 1.873%), 1.168% (0.429%, 1.907%), 0.655% (0.283%, 1.652%). The protein expression of Beclin-1 in the tissue of rats where the Freund′s complete adjuvant was injected in BCG+ rapamycin group was significantly higher than that of blank control group (

=4.796,

<0.05). The protein expression of LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in BCG alone group was significantly higher than that of blank control group (

=4.953,

<0.05). Conclusions Freund′s complete adjuvant can enhance the expression levels of local tissue autophagy-related proteins Beclin-1 and LC3B in rat tuberculous wound model.

Objective To investigate the expression and effect of microRNA-205 (miR-205) in human hypertrophic scar. Methods The experimental research method was applied. From October 2019 to January 2020, hypertrophic scar tissue from 6 patients with hypertrophic scar (1 male and 5 females, aged (36±7) years) and remaining normal skin tissue from 6 trauma patients (2 males and 4 females, aged (38±9) years) after flap transplantation operation were collected. The above-mentioned 12 patients were admitted to the General Hospital of Northern Theater Command and met the inclusion criteria. Real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to detect the mRNA expressions of miR-205 and thrombospondin-1 (TSP-1). The hypertrophic scar tissue was taken to culture the 3rd to 5th passage of fibroblasts (Fbs) for the follow-up experiments. Two batches of hypertrophic scar Fbs were divided into TSP-1+ miR-205 control group, TSP-1+ miR-205 mimic group, and TSP-1 mutant+ miR-205 control group, TSP-1 mutant+ miR-205 mimic group, which were transfected with the corresponding sequences. At 48 h after transfection, the expressions of luciferase and renal luciferase were detected by luciferase reporter gene detection kit, and the luciferase/renal luciferase ratio was calculated to indicate the activity of TSP-1. Two batches of hypertrophic scar Fbs were collected and divided into miR-205 control group, miR-205 mimic group, and miR-205 inhibitor group and miR-205 control group, miR-205 mimic group, and miR-205 mimic+ TSP-1 group, respectively, which were transfected with the corresponding sequences. At 0 (immediately), 12, 24, 36, and 48 h after transfection, the cell viability was detected by microplate reader. Two batches of hypertrophic scar Fbs were grouped and treated as described in the cell viability detecting experiment. At 24 h after transfection, Hoechst 33258 staining was performed to observe the nuclear shrinkage, so as to reflect the apoptosis of Fbs. The number of samples in cell experiment was three. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance,

test, and chi-square test. Results The mRNA expression of miR-205 in hypertrophic scar tissue was 0.54±0.05, which was significantly lower than 1.26±0.07 in normal skin tissue (

=8.213,

<0.01). The expression of TSP-1 mRNA in hypertrophic scar tissue was 1.46±0.07, which was significantly higher than 0.68±0.11 in normal skin tissue (

=6.031,

<0.01). At 48 h after transfection, the luciferase/renal luciferase ratio reflecting the TSP-1 activity of cells in TSP-1+ miR-205 mimic group was 0.532±0.028, which was significantly lower than 0.998±0.012 in TSP-1+ miR-205 control group (

=26.500,

<0.01), and the luciferase/renal luciferase ratio of cells in TSP-1 mutant+ miR-205 mimic group was 0.963±0.012, which was close to 0.976±0.010 in TSP-1 mutant+ miR-205 control group (

=0.816,

>0.05). At 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 mimic group was significantly lower than that in miR-205 control group (

=6.169, 12.670, 27.130, 12.670,

<0.05 or

<0.01). At 0, 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 inhibitor group was significantly higher than that in miR-205 control group (

=6.169, 7.221, 7.787, 7.835, 13.030,

<0.05 or

<0.01). At 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 mimic group was significantly lower than that in miR-205 control group and miR-205 mimic+ TSP-1 group (

=8.118, 26.970, 39.550, 42.490, 14.570, 12.240, 36.830, 45.220,

<0.05 or

<0.01). At 24 h after transfection, compared with miR-205 control group, the cell apoptosis in miR-205 mimic group was increased, and the cell apoptosis in miR-205 inhibitor group was decreased. At 24 h after transfection, compared with miR-205 mimic group, the cell apoptosis in miR-205 control group and miR-205 mimic+ TSP-1 group were decreased. Conclusions miR-205 can inhibit the proliferation and promote the apoptosis of Fbs in human hypertrophic scar by inhibiting the expression of TSP-1, which has the potential to be a therapeutic target for hypertrophic scar.

Objective To explore the clinical effects of anterograde sural neurovascular flap in repairing skin and soft tissue defect around the knee. Methods Nine patients with skin and soft tissue defect around the knee admitted to Beijing Fengtai YouAnMen Hospital from May 2011 to December 2018, were included in this retrospective descriptive study, including 8 males and 1 female, aged 16 to 65 years. The wound area after debridement ranged from 8 cm×5 cm to 18 cm×10 cm. Anterograde sural neurovascular flap was used to repair the wounds in 9 patients, with the area ranging from 9 cm×6 cm to 20 cm×12 cm. The donor sits of flaps in 2 patients were closed and sutured directly, and the donor sits of flaps in 7 patients were repaired with medial split-thickness skin graft of the ipsilateral thigh. The flap survival, complications, and follow-up after operation were recorded. Results The flaps survived and the blood supply was good in 8 patients and the wounds were closed. One patient developed skin ischemic necrosis which was cured after three weeks of dressing change. All the skin grafts in the donor site of flap in 7 patients survived. In 6 months to 5 years of follow-up after surgery, the skin flap had good texture, color, and shape, and normal sensation. Except for one patient whose knee had poor recovery of function, the knee joint function of the other patients recovered well. Conclusions The anterograde sural neurovascular flap has the advantages of high survival rate, satisfactory appearance and functional recovery post surgery, and is an ideal flap for repairing the skin and soft tissue defect around the knee.