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2019 Vol. 35, No. 1

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A Letter from the Editor-in-Chief
Set sail again and look forward to new 60 years
Peng Yizhi
2019, 35(1): 1-2. doi: 10.3760/cma.j.issn.1009-2587.2019.01.001
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The outstanding work of Chinese Journal of Burns in 2018: the successful planning and display of the " Burn Medicine Over the Past 60 Years" column, the launch of seven guidelines and consensus for burn injury treatment, and the opening of the " Online First" and " Video Center" sections in the journal website. Specifically, the " Burn Medicine Over the Past 60 Years" celebration column invited more than 30 papers with sincerity from academicians, seniors in the burn academic field, and listed departments in Fudan University′s academic ranking as representatives to praise 60 years discipline development history and exchange the future construction and development concepts. We hope that the fellows enjoy and ponder over the discussions to explore new academic journeys. Our journal, with firm hearts, will accompany you to work together and accomplish new peaks in burn filed.
Academician Forum
Role of cytokines in sepsis and its current situation of clinical application
Xia Zhaofan, Wu Guosheng
2019, 35(1): 3-7. doi: 10.3760/cma.j.issn.1009-2587.2019.01.002
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Sepsis is defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection, which is a global health crisis. The cytokines are a class of protein mediators secreted by cells with low molecular weights and biological activity. There are two types of cytokines, pro-inflammatory and anti-inflammatory cytokines, participating in regulation of immunity and inflammation in sepsis. Cytokines are also used as biomarkers for early warning, diagnosis, and prognostic assessment of sepsis and as therapeutic targets for prevention and treatment of sepsis. This paper briefly summarizes the research advance of some cytokines in sepsis and the related work carried out by author′s institute, and elaborates the roles of cytokines in sepsis and their clinical application value, helping to provide some ideas and reference for the future research of cytokines in sepsis.
Expert Forum
Developing trend of wound dressing
Huan Jingning
2019, 35(1): 8-11. doi: 10.3760/cma.j.issn.1009-2587.2019.01.003
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The ideal wound dressing should have the functions of keeping wound moist and warm, preventing and treating wound infection, promoting wound healing, and so on; However there is no such ideal wound dressing in clinic. Dressings are likely to capable application to different kinds of wounds with multi-functions in the future. For the purpose of good tissue compatibility and permanent wound cover, auto- or allo- skin living cells should be integrated with biological dressings as real artificial skin by employing tissue engineering technology. Clinical application of smart dressings can enable wound management more personalized, effective, optimized, and convenient.
Original Article·New Technology and Dressing of Wound Treatment
Content characteristics and cytotoxicity detection of silver ion products
Su Xuerong, Gan Junying, Xue Yuying, Li Qiang
2019, 35(1): 12-17. doi: 10.3760/cma.j.issn.1009-2587.2019.01.004
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Objective To analyze the silver content, homogeneity, and cytotoxicity of silver-containing products. Methods (1) Five kinds of silver-containing products A, B, C, D, and E were purchased from the market, and products A, B, C, and D are liquid or gel form while product E was dressing form. The silver content of each product and the homogeneity of product E were determined by flame method. The sample number was 3. (2) Human hepatocellular carcinoma cell line (HepG2) was selected as the evaluation model. Four silver-containing products A, B, C, and D were diluted with high-glucose dulbecco′s modified eagle medium (DMEM) at multiple ratios of 1∶100, 1∶200, 1∶400, and 1∶800, and then they were used for cell culture. Cells cultured with high-glucose DMEM and high-glucose DMEM containing 20 μg/mL silver nitrate were used as blank control and positive control, respectively. The cell viability was determined by methyl thiazolyl tetrazolium assay, and each sample number was 5. (3) Four mass concentrations of 0.031 3, 0.062 5, 0.125 0, and 0.250 0 μg/mL were prepared from silver-containing product A, and then they were used to culture HepG2 cell. Cells cultured with high-glucose DMEM containing fetal calf serum and 294 μg/mL potassium dichromate were used as positive control, while those containing fetal calf serum were used as blank control. Hoechst 33258 staining method was used to detect apoptosis rate of cells. The tail moment, tail length, and the percentage of DNA in the tail of cells were observed by comet assay to evaluate DNA damage. The sample numbers were all 3. Data were processed with one-way analysis of variance and least significant difference-t test. Results The silver content of products A, B, C, and D was (256.5±1.5) μg/mL, (271.5±1.3) μg/mL, (652.4±2.6) μg/g , (330.0±2.1) μg/g, which was in accordance with labelled amount. The silver content of product E was (0.158±0.013) mg/g, and the silver content of each piece of product E was (0.125±0.017) mg/g, showing good uniformity of product E. (2) Compared with the rate of blank control, the cell survival rates of product A at the dilution ratio of 1∶100, product B at the dilution ratio of 1∶100, and product C at the dilution ratio of 1∶100 and 1∶200 were significantly reduced (t=35.506, 8.914, 37.594, 30.693, P<0.01). Compared with the rate of positive control, the cell survival rates of product A at the dilution ratio of 1∶200, 1∶400, and 1∶800, product C at the dilution ratio of 1∶400 and 1∶800, products B and D at each dilution ratio were increased significantly (t=27.537, 18.262, 18.709, 26.333, 41.762, 15.776, 19.759, 20.443, 15.715, 26.792, 24.963, 31.803, 30.537, P<0.01). (3) The apoptosis rates of cells treated by 0.250 0 μg/mL product A and positive control were (6.1±0.4)% and (62.2±3.9)% respectively, which were significantly higher than the apoptosis rate of blank control [(3.3±0.7)%, t=13.327, 30.475, P<0.05]. The apoptosis rates of cells treated by 0.031 3, 0.062 5, 0.125 0 μg/mL product A were (2.9±0.4)%, (3.1±0.4)%, and (4.2±0.9)% respectively, which were close to the apoptosis rate of blank control (t=1.181, 0.133, 1.097, P>0.05). (4) The tail moment, tail length, and tail DNA percentage of cells cultured with 0.125 0 and 0.250 0 μg/mL product A were significantly higher than those cultured with blank control (t=29.026, 51.194, 21.851, 36.138, 24.721, 50.455, P<0.05 or P<0.01). However, the tail moment, tail length, and tail DNA percentage of cells cultured with 0.031 3 and 0.062 5 μg/mL product A were close to those cultured with blank control (t=5.878, 3.429, 2.779, 1.960, 1.328, 7.763, P>0.05). Conclusions The silver content of silver-containing products meets the requirements of the labeling. The concentration of product C is higher than that of other products, leading to a greater possibility of decreasing the survival rate of HepG2 cells. It is suggested that the products A and B should be taken as reference in the concentration setting of silver ion products. The product solution with higher concentration may have higher risk of damage to cell DNA. Therefore, it is not recommended to upregulate silver content of relevant products blindly in order to achieve better antibacterial effect.
Analysis of differential gene expressions of inflammatory and repair-related factors in chronic refractory wounds in clinic
Wang Lian, Guo Fei, Min Dinghong, Liao Xincheng, Yu Shaoqing, Long Xingxing, Ding xiang, Guo Guanghua
2019, 35(1): 18-24. doi: 10.3760/cma.j.issn.1009-2587.2019.01.005
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Objective To compare the tissue morphology and gene expressions of inflammatory and repair-related factors in chronic refractory wound tissue including pressure ulcers and diabetic feet. Methods During August 2016 to September 2017, 10 samples of prepuce were collected after circumcision of 10 urological patients [all male, aged (38±4) years old] admitted in the First Affiliated Hospital of Nanchang University and included in normal skin group, samples of tissue around the edge of wounds with blood supply were collected from 9 heat or electric burn patients [6 male patients, 3 female patients, aged (51±8) years old], 13 pressure ulcer patients [9 male patients, 4 female patients, aged (51±14) years old] and 10 diabetic foot patients [8 male patients, 2 female patients, aged (61±10) years old] during the operations. The samples were divided into burn wound group (9 samples), pressure ulcer group (13 samples), and diabetic foot group (10 samples). Ten slices were taken from pressure ulcer group and diabetic foot group respectively, and 5 slices in each group were used to observe the tissue morphology and expressions of Ki67 and CD31 of wounds respectively with immunofluorescence method. Ten samples from normal skin group, 9 samples from burn wound group, 13 samples from pressure ulcer group, and 10 samples from diabetic foot group were collected for analysis of mRNA expressions of vascular endothelial growth factor 192 (VEGF192), transforming growth factor β (TGF-β), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) , interleukin-1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α) by real time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with Mann-Whitney U test and Kruskal-Wallis rank-sum test. Results (1) The expression level of Ki67 in diabetic foot group (390±100) was higher than that of pressure ulcer group (182±14, Z=-2.611, P<0.01). (2) Although there were a large number of vascular endothelial cells (CD31 positive cells) in wounds of diabetic foot group, their distribution was disordered and failed to form intact lumen. There were less vascular endothelial cells in wounds of pressure ulcer group than those of diabetic foot group, but the complete lumen was formed. (3) The mRNA expression levels of VEGF192 in wounds of burn wound group, pressure ulcer group, and diabetic foot group were significantly lower than the level in normal skin group (H=13.72, 30.50, 15.20, P<0.05 or P<0.01), and the level was the lowest in pressure ulcer group. The mRNA expression level of VEGF192 in wounds of pressure ulcer group was significantly lower than that of diabetic foot group (H=15.30, P<0.01). Compared with that of normal skin group, the mRNA expression level of TGF-β in wounds of burn wound group showed no significant difference (H=-9.50, P>0.05), while the mRNA expression levels of TGF-β in wounds of pressure ulcer group and diabetic foot group were significantly decreased (H=18.04, 14.50, P<0.01). The mRNA expression level of TGF-β in wounds of pressure ulcer group was similar to that of diabetic foot group (H=3.54, P>0.05). (4) Compared with those of normal skin group, the mRNA expression levels of VCAM-1 in wounds of burn wound group and pressure ulcer group were significantly increased (H=-22.50, -11.50, P<0.05 or P<0.01), and there was no significant difference in the mRNA expression level of VCAM-1 in wounds of diabetic foot group (H=10.00, P>0.05); the mRNA expression level of ICAM-1 in wounds of burn wound group showed no significant difference (H=-9.50, P>0.05), and the levels of ICAM-1 in wounds of pressure ulcer group and diabetic foot group were significantly decreased (H=16.50, 16.50, P<0.01). The mRNA expression level of VCAM-1 in wounds of pressure ulcer group was significantly higher than that of diabetic foot group (H=-21.50, P<0.01), the mRNA expression level of ICAM-1 in wounds of pressure ulcer group was similar to that of diabetic foot group (H=0, P>0.05). (5) Compared with those of normal skin group, except for the mRNA expression level of IL-1β in wounds of diabetic foot group showed no significant difference (H=-10.00, P>0.05), the mRNA expression levels of IL-1β in wounds of burn wound group and pressure ulcer group were significantly increased (H=-32.50, -21.50, P<0.01); the mRNA expression levels of IL-6 were significantly increased in wounds of burn wound group, pressure ulcer group, and diabetic foot group (H=-17.50, -30.50, -11.80, P<0.05 or P<0.01); except for the mRNA expression level of TNF-α in wounds of burn wound group showed no significant difference (H=-9.50, P>0.05), the mRNA expression levels of TNF-α in wounds of pressure ulcer group and diabetic foot group were significantly decreased (H=18.04, 14.50, P<0.01). The mRNA expression levels of IL-1β and TNF-α in wounds of pressure ulcer group were significantly lower than those of burn wound group (H=11.00, 27.54, P<0.05 or P<0.01), while the mRNA expression level of IL-6 was significantly higher (H=-13.00, P<0.05). The mRNA expression levels of IL-1β and TNF-α in wounds of diabetic foot group were significantly lower than those of burn wound group (H=22.50, 24.00, P<0.01), while the mRNA expression level of IL-6 showed no significant difference (H=5.70, P>0.05). Conclusions The phenotypes of diabetic foot and pressure ulcer vary from the expressions levels of proliferating cell nuclear antigen and blood vessels forming ability to the expression levels of growth factors, cell adhesion factors, and inflammatory cytokines.
Clinical characteristics and treatment of diabetic patients with superficial partial-thickness burn on feet
Ling Xiangwei, Zhang Tingting, Dai Wentong, Xia Weidong, Lin Cai
2019, 35(1): 25-30. doi: 10.3760/cma.j.issn.1009-2587.2019.01.006
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Objective To analyze the characteristics and treatment of diabetic patients with superficial partial-thickness burn on feet. Methods Eighty-three patients with superficial partial-thickness burn on 119 feet were hospitalized in our unit from January 2011 to December 2017. The medical records of the patients with 46 men and 37 women, aged 60±11 were retrospectively analyzed. The patients were divided into diabetes group and non-diabetes group according to whether they had diabetes or not, with 41 patients (60 burn feet) in diabetes group and 42 patients (59 burn feet) in non-diabetes group. Patients in diabetes group and non-diabetes group were given systemic treatment and wound dressing change. Thirty-seven diabetic patients whose wounds deepened to deep partial-thickness were divided into eschar shaving group and non-eschar shaving group according to patients′ willingness and the treatment, with 14 patients in eschar shaving group and 23 patients in non-eschar shaving group. Patients in eschar shaving group were given eschar shaving operation at early stage, and patients in non-eschar shaving group were given wound dressing change. The length of hospital stay, hospitalization treatment expenses, pulse of arteria dorsal pedis and posterior tibial artery immediately after admission, deepening of wounds on feet during hospital stay, and rates of wound healing on feet of patients in diabetes group and non-diabetes group were observed and calculated. Pulses of arteria dorsal pedis and posterior tibial artery immediately after admission, deepening of wounds on feet during hospital stay, positive rates of bacteria and fungus in wounds on feet, and rates of wound healing on feet of patients in eschar shaving group and non-eschar shaving group were observed and calculated. Data were processed with chi-square test, t test, Fisher′s exact propability method, and Mann-Whitney U test. Results The length of hospital stay of patients in diabetes group was (29±20) d, which was significantly longer than that of patients in non-diabetes group [(19±13) d, t=2.730, P<0.01]. The hospitalization treatment expense of patients in diabetes group was (46 988±41 322) yuan, which was significantly more than that of patients in non-diabetes group [(29 106±24 813) yuan, t=2.396, P<0.05]. The pulses of arteria dorsal pedis and posterior tibial artery of patients in diabetes group were significantly weaker than those of patients in non-diabetes group (Z=3.278, 2.194, P<0.05 or P<0.01). The percentages of wounds on feet of patients in diabetes group deepening to deep partial-thickness burn, full-thickness skin defect with bone and tendon exposure were respectively 88.3% (53/60) and 23.3% (14/60), which were significantly higher than those of patients in non-diabetes group [47.5% (28/59) and 1.7% (1/59), χ2=22.867, 12.644, P<0.01]. Rate of wound healing on feet of patients in diabetes group was 78.3% (47/60), which was significantly lower than 100.0% (59/59) of patients in non-diabetes group ( χ2=14.351, P<0.01). There were respectively 21 and 32 feet in patients of eschar shaving group and non-eschar shaving group. There were no significantly statistical differences in pulses of arteria dorsal pedis and posterior tibial artery of patients between eschar shaving group and non-eschar shaving group (Z=0, 0.453, P>0.05). The percentage of wounds on feet of patients in non-eschar shaving group deepening to full-thickness skin defect with bone and tendon exposure was 43.8% (14/32), which was significantly higher than 0 of patients in eschar shaving group ( χ2=12.486, P<0.01). Positive rates of bacteria and fungus in wounds on feet of patients in eschar shaving group was significantly lower than that of patients in non-eschar shaving group (χ2=4.386, P<0.05 ). Rate of wound healing on feet of patients in non-eschar shaving group was 59.4% (19/32), which was significantly lower than that of patients in eschar shaving group [100.0% (21/21), P<0.01]. Conclusions Diabetes patients with superficial partial-thickness burn wounds on feet has long length of hospital stay, high hospitalization treatment expenses. Wounds of the patients are easy to deepen, with low wound healing rate. Eschar shaving at early stage when the wounds deepened to deep partial-thickness burn is a good way to increase wound healing rate and prevent further deepening of wounds.
Effects of platelet-rich plasma combined with polylactic acid/polycaprolactone on healing of pig deep soft tissue defect caused by fragment injury
Yao Dan, Hao Daifeng, Zhao Fan, Hao Xiufeng, Feng Guang, Chu Wanli, Chen Zequn
2019, 35(1): 31-39. doi: 10.3760/cma.j.issn.1009-2587.2019.01.007
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Objective To investigate the effects of platelet-rich plasma (PRP) combined with polylactic acid/polycaprolactone (PLA/PCL) on healing of mininature pig deep soft tissue defect caused by fragment injury. Methods Two male Bama miniature pigs with 11 to 12 months (the same below) were selected by lottery to prepare PRP. The other twenty-seven male Bama miniature pigs were used to reproduce deep soft tissue defect caused by high-explosive ammunition fragment injury on bilateral posterior femoral region. According to the random number table, 27 pigs were divided into control group, material group, and PRP+material group, with 9 pigs in each group. After debridement, wounds of pigs in material group and PRP+material group were filled with PLA/PCL and PLA/PCL+2 mL activated PRP, respectively. Pigs in each group received suture of full-thickness skin to close the wounds. The operative duration was recorded. The length and volume of wounds of pigs in the above groups were measured immediately after surgery. In 1, 2, and 4 weeks after surgery, 3 pigs in each group were sacrificed to collect femoral wounds tissue on two sides, and PLA/PCL were collected from wounds of pigs in material group and PRP+material group for general observation of wounds tissue and degradation of the material. In 2 and 4 weeks after surgery, wounds tissue was obtained to observe the histological changes by hematoxylin-eosin staining, and expressions of transforming growth factor β (TGF-β) and vascular endothelial growth factor (VEGF), and angiogenesis were determined by immunohistochemical method. In 1, 2, and 4 weeks after surgery, wounds tissue was collected to determine mRNA expressions of TGF-β and VEGF by real-time quantitative reverse transcription polymerase chain reaction. Data were processed with analysis of variance of factorial design, one-way analysis of variance, and least significant difference-t test. Results (1) There were no significantly statistical differences in length and volume of the wounds of pigs among the three groups (F=0.336, 0.282, P>0.05). The operative duration in control group [(30.9±2.1)min] was significantly shorter than that of material group [(39.7±2.2)min] and PRP+material group[(40.0±2.6)min], t=-11.45, -11.88, P<0.01. (2) There were respectively 10, 7, and 5 wounds tissue with infection in pigs of control group, material group, and PRP+material group. In 1, 2, 4 weeks after surgery, all of the wounds tissue of pigs was infected in control group, while none of wounds tissue of pigs was infected in material group and PRP+material group. In pigs of material group and PRP+material group, materials and tissue were easily separated in 1 week after surgery; some materials were integrated with tissue and showed a tendency of degradation in 2 weeks after surgery; materials were completely embedded with tissue in 4 weeks after surgery. (3) In pigs of control group, erythrocytes and inflammatory cells infiltration in wounds tissue were observed in 2 weeks after surgery, and necrotic tissue and inflammatory cells infiltration in wounds tissue were still observed in 4 weeks after surgery. In pigs of material group and PRP+material group, a large number of erythrocytes and inflammatory cells infiltration were observed in 2 weeks after surgery. Compared with that of material group, wounds tissue of pigs in PRP+material group had no inflammatory cells infiltration in 4 weeks after surgery. (4) Protein expressions of TGF-β in fibroblasts and multinuclear macrophagocytes, VEGF in fibroblasts and vascular endothelial cells, and blood vessel formation in wounds tissue of pigs in PRP+material group were significantly more than those of pigs in control group and material group in 2 and 4 weeks after surgery. (5) The mRNA expression of TGF-β in wounds tissue of pigs in material group was significantly higher than that in control group in 4 weeks after surgery (t=-3.93, P<0.01). Compared with those of pigs in control group and material group, the mRNA expression of TGF-β in wounds tissue of pigs in PRP+material group was significantly increased at each time point (t=9.23, 13.81, 11.73, -7.51, -12.04, -7.80, P<0.01). The mRNA expression of VEGF in wounds tissue of pigs increased significantly in material group compared with that of pigs in control group in 4 weeks after surgery (t=-3.94, P<0.01). Compared with those of pigs in control group and material group, the mRNA expression of VEGF in wounds tissue increased significantly in wound tissue of pigs in PRP+material group at each time point (t=12.33, 3.95, 7.97, -11.36, -2.97, -4.04, P<0.01). Conclusions PRP combined with PLA/PCL can accelerate wound healing of deep soft tissue defect of mininature pigs caused by fragment injury by providing physical scaffold for newborn tissue growth, promoting mRNA and protein expressions of TGF-β and VEGF.
Effects of adipose-derived mesenchymal stem cells from type 2 diabetes mellitus patients on wound healing of pressure ulcers in mice
Deng Chengliang, Yao Yuanzhen, Liu Zhiyuan, Wang Bo, Wang Dali, Wei Zairong
2019, 35(1): 40-47. doi: 10.3760/cma.j.issn.1009-2587.2019.01.008
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To investigate the effects of adipose-derived mesenchymal stem cells (AMSCs) from type 2 diabetes mellitus patients on wound healing of pressure ulcers in mice. Methods (1) In September 2016, the subcutaneous adipose tissue of a 60-year-old woman with type 2 diabetes mellitus was harvested, and then AMSCs were extracted by collagenase digestion and cultured. The third passage of cells were used for subsequent experiments. The morphology of cells was observed, and their osteogenic, chondrogenic, and adipogenic differentiation abilities were identified. The expressions of cell surface markers CD90, CD105, CD73, and CD34 were detected by flow cytometer (n=3). (2) Sixteen female C57BL/6 wild-type mice aged 6-8 weeks were selected, and one pressure ulcer wound was created on each side of the spine of each mouse by pressing the skin with two magnets. The two wounds of each mouse were paired and divided into diabetic AMSCs group and negative control group, injected with 100 μL phosphate buffer solution (PBS) containing green fluorescent protein-labeled AMSCs (1×106 cells) and 100 μL PBS, respectively. The wound healing status of the two groups within post injection day (PID) 21 was observed, and their wound healing rates on PID 5, 13, and 17 were calculated. Three mice were sacrificed on PID 11 and 21, respectively, and tissue of three wounds was harvested from each group. The skin structure was observed by hematoxylin-eosin staining, the collagen deposition was evaluated by Masson staining, and the positive expression of CD31, i. e., the number of new blood vessels was counted by immunohistochemistry. Wound tissue samples of two groups prepared on PID 21 as above-mentioned were harvested, and the positive cell rate of S100, representing the regeneration of Schwann cells, was detected by immunohistochemistry. Wound tissue samples of diabetic AMSCs group prepared on PID 11 as above-mentioned were harvested, and the colonization of AMSCs was observed by fluorescence tracer method. Data were processed with paired t test and Bonferroni correction. Results (1) The third passage of cells isolated and cultured from the subcutaneous adipose tissue of a type 2 diabetes mellitus patient grew adherently to the wall in a long spindle and vortex-like manner. After induction, the cells showed osteogenic, chondrogenic, and lipogenic differentiation abilities. The positive expression rates of CD90, CD105, and CD73 on the cell surface were higher than 90.00%, and the expression rate of CD34 was 0.46%. The cells were identified as AMSCs. (2) The mice wounds of diabetic AMSCs group healed quickly, and all the wounds healed completely on PID 17, while the mice wounds in negative control group were not completely closed at this time, and there was still scab on the surface. On PID 5, 13, and 17, the healing rates of mice wounds of diabetic AMSCs group were (35.6±6.5)%, (87.1±2.5)%, and 100.0%, respectively, significantly higher than (19.8±7.2)%, (66.2±5.2)%, and (86.9±5.3)% of negative control group (t=6.49, 14.31, 9.73, P<0.05). Compared with that of negative control group, the inflammatory cell infiltration was reduced in mice wounds tissue of diabetic AMSCs group on PID 11, and thicker epidermis and dermis as well as regenerated skin appendages were observed on PID 21. On PID 11 and 21, the collagen percentages of mice wounds tissue in diabetic AMSCs group was (48.3±1.3)% and (54.1±1.7)%, respectively, significantly higher than (41.4±1.7)% and (50.3±1.2)% of negative control group (t=6.98, 3.99, P<0.01). On PID 11 and 21, the numbers of new blood vessels in mice wounds tissue of diabetic AMSCs group were 17.2±1.3 and 18.0±2.1, respectively, significantly more than 8.0±1.4 and 14.0±1.5 of negative control group (t=10.69, 3.38, P<0.01). On PID 21, the S100 positive cell percentage in mice wounds tissue of diabetic AMSCs group was (1.76±0.12)%, significantly higher than (0.55±0.03)% of negative control group (t=21.68, P<0.001). On PID 11, the colonization of AMSCs in mice wounds tissue of diabetic AMSCs group was observed. Conclusions Transplantation of AMSCs from type 2 diabetic mellitus patients can accelerate wound healing of pressure ulcers in mice by promoting angiogenesis, collagen deposition, and Schwann cell regeneration.
Original Article
Effects of platelet-rich plasma on the survival of ultra-long dorsal random flaps in rats
Tian Xinli, Jiang Bo, Yan Hong
2019, 35(1): 48-53. doi: 10.3760/cma.j.issn.1009-2587.2019.01.009
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Objective To observe the effects of platelet-rich plasma (PRP) on the survival of ultra-long dorsal random flaps in rats. Methods Sixteen male Sprague Dawley rats aged 6 to 8 weeks (the same below) were sacrificed to collect whole blood of 9 to 10 mL from each rat, and PRP was prepared by modified APPLE method. The platelet count of retained whole blood and PRP detected by automated blood cell analyzer showed that PRP was made successfully. The other thirty-two rats were collected and divided into PRP group and control group according to the random number table, with 16 rats in each group. One rectangular ultra-long random flap with area of 8 cm×2 cm was made on the back of each rat and replanted in situ. The equidistant 3 points were designed on both sides of the flap of each rat. Rats in PRP group were injected with 0.1 mL PRP from dermis and subcutaneous tissue of each injection point, while rats in control group were injected with the same volume of normal saline. Eight rats in each group were sacrificed at post operation hour (POH) 24 and on post operation day (POD) 7. On POD 7, survival of flaps of rats in 2 groups was observed, and the survival rates of flaps were calculated. On POD 7, the proximal, middle, and distal flaps of rats in 2 groups were collected, and histological changes of the flaps of rats in 2 groups were observed with hematoxylin-eosin staining. At POH 24 and on POD 7, flaps in 3 to 4 cm to pedicles were taken to detect mRNA expressions of vascular endothelial growth factor (VEGF), platelet-derived growth factor AA (PDGF-AA) and PDGF-BB by real-time fluorescent quantitative reverse transcription polymerase chain reaction, and to determine content of nitric oxide by nitrate reductase method. Data were processed with t test. Results (1) On POD 7, flaps of rats in PRP group were dry without purulent exudate, and covered with scab, and the pink new skin emerged after scab fell off. On POD 7, flaps of rats in control group were with a large amount of inflammatory exudates, 1/2 to 2/3 of flaps at the distal were with necrosis and covered by scab which was not easy to be stripped. The survival rate of flap of rats in PRP group was (67±6)%, significantly higher than (52±10)% of rats in control group (t=1.94, P<0.05). (2) There were no obvious inflammatory cell infiltration and a number of microvessels, and fibrous tissue arranged neatly in the proximal flaps of rats in PRP group. There were a few of inflammatory cell infiltration and a number of microvessels, and fibrous tissue arranged slightly disorderly in the middle flaps of rats in PRP group. There were many more inflammatory cell infiltration and microvessels, a small amount of vascular embolism, and fibrous tissue arranged disorderly in the distal flaps of rats in PRP group. There were a large number of inflammatory cells infiltration and a few of microvessels, and fibrous tissue arranged disorderly in the proximal, middle, and distal flaps of rats in control group. (3) At POH 24 and on POD 7, mRNA expressions of VEGF, PDGF-AA, and PDGF-BB of rats in PRP group were significantly higher than those of rats in control group (t=6.46, 5.61, 2.88, 10.18, 6.10, 7.67, P<0.001). (4) At POH 24, content of nitric oxide in flap of rats in PRP group was (5.0±0.9) μmol/g, significantly higher than (3.4±0.9) μmol/g of rats in control group (t=19.14, P<0.001). On POD 7, content of nitric oxide in flap of rats in PRP group was (3.3±0.8) μmol/g, which was close to (3.0±0.6) μmol/g of rats in control group (t=2.93, P>0.05). Conclusions PRP can improve the survival rate of ultra-long dorsal random flap in rats, which may be related to regulation of angiogenesis related factors, increase of nitric oxide content, and inhibition of excessive apoptosis of cells of PRP, so as to alleviate ischemical reperfusion injury and improve microcirculatory disturbance.
Protective effects and mechanism of keratinocyte growth factor combined with hypoxia inducible factor-1α on intestinal crypt epithelial cells of rats with hypoxia stress
Xu Qian, Bai Yanqing, Zeng Tongxu, Yang Bo, Cai Xiaoling, Ha Xiaoqin
2019, 35(1): 54-61. doi: 10.3760/cma.j.issn.1009-2587.2019.01.010
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Objective To investigate the protective effects and mechanism of keratinocyte growth factor (KGF) combined with hypoxia inducible factor-1α (HIF-1α) on intestinal crypt epithelial cells (IEC-6) of rats with hypoxia stress. Methods (1) The routinely cultured IEC-6 of rats were collected and divided into normoxia blank group, normoxia KGF group, normoxia HIF-1α group, and normoxia combine group, according to the random number table, and then the previous mediums were respectively replaced with dulbecco′s modified eagle medium (DMEM), medium with 0.5 ng/mL KGF, medium with 10.0 ng/mL HIF-1α, and medium with 0.5 ng/mL KGF and 30.0 ng/mL HIF-1α. And the cells were cultured in cell incubator with oxygen volume fraction of 21% for 24 hours. (2) Another batch of routinely cultured IEC-6 were collected and divided into normoxia control group, hypoxia control group, hypoxia KGF group, hypoxia HIF-1α group, and hypoxia combine group, according to the random number table. The previous mediums were replaced with DMEM, DMEM, medium with 0.5 ng/mL KGF, medium with 10.0 ng/mL HIF-1α, and medium with 0.5 ng/mL KGF and 30.0 ng/mL HIF-1α respectively. And then, the cells in normoxia control group were cultured routinely for 24 hours, and cells in the other 4 groups were cultured in cells incubator of 3 gases, with oxygen volume fraction of 5% for 24 hours. Cells cultured in normoxic and hypoxic incubators were collected, with 3 samples in each group, and morphological changes of cells were observed with optical microscope. Cells cultured in normoxic and hypoxic incubators were collected, with 3 samples in each group, and survival rates of cells were detected by cell count kit 8. Cells in normoxia control group and cells cultured in hypoxic incubator were collected, with 3 samples in each group. The cell cycle changes and apoptosis rates were detected by flow cytometer, the content of adenosine triphosphate (ATP) was detected by ultraviolet spectrophotometer, and protein expression of p53 was detected by Western blotting. Data were processed with one-way analysis of variance and least significant difference test. Results (1) After being cultured for 24 h, cells cultured in normoxic incubator grew well with oval or round shapes and clear cytoplasm, and cells cultured in hypoxic incubator showed irregular shapes such as fusiform or starlike shape, with black particle in cytoplasm. (2) After being cultured for 24 h, cell survival rates of normoxia blank group, normoxia KGF group, normoxia HIF-1α group, and normoxia combine group were (107.4±8.7)%, (109.8±2.9)%, (115.8±7.4)%, and (112.8±10.6)% respectively. There was no significantly statistical difference in general comparison of cell survival rates among the above groups (F=0.685, P=0.586). After being cultured for 24 h, cell survival rates of hypoxia control group, hypoxia KGF group, hypoxia HIF-1α group, and hypoxia combine group were (35.1±4.6)%, (52.9±6.8)%, (56.2±3.1)%, and (71.2±9.6)% respectively, which were significantly lower than (106.3±12.3)% of normoxia control group (P<0.001). Survival rates of cells in hypoxia KGF group, hypoxia HIF-1α group, and hypoxia combine group were significantly higher than the rate of cells in hypoxia control group (P=0.023, 0.009, <0.001). Survival rate of cells in hypoxia combine group was significantly higher than the rates of cells in hypoxia KGF group and hypoxia HIF-1α group (P=0.017, 0.045). (3) After being cultured for 24 h, percentage of cells in G1 phase in hypoxia control group was significantly higher than that of cells in normoxia control group (P=0.030), percentages of cells in S phase in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group were obviously lower than the percentage of cells in normoxia control group (P=0.020, 0.031, 0.026), and percentages of cells in different phases in other groups were close to those of cells in normoxia control group (P=0.516, 0.107, 0.052, 0.985, 0.637, 0.465, 0.314, 0.591). After being cultured for 24 h, percentages of cells in G1 phase in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group were obviously higher than the percentage of cells in hypoxia combine group (P=0.001, 0.030, 0.014), and percentages of cells in S phase in the above 3 groups were obviously lower than the percentage of cells in hypoxia combine group (P=0.001, 0.012, 0.010). (4) After being cultured for 24 h, compared with that of cells in normoxia control group, apoptosis rate of cells in hypoxia control group obviously increased (P=0.018), and apoptosis rate of cells in hypoxia combine group obviously decreased (P=0.008). After being cultured for 24 h, compared with that of cells in hypoxia control group, apoptosis rates of cells in hypoxia KGF group and hypoxia combine group obviously decreased (P=0.004, 0.001). Apoptosis rate of cells in hypoxia combine group was obviously lower than those of cells in hypoxia KGF group and hypoxia HIF-1α group (P=0.032, 0.002). (5) After being cultured for 24 h, compared with that of cells in normoxia control group, the content of ATP of cells in hypoxia combine group changed unobviously (P=0.209), and content of ATP of cells in the other groups obviously decreased (P= <0.001, 0.001, 0.002). Content of ATP of cells in hypoxia HIF-1α group and hypoxia combine group was obviously higher than that of cells in hypoxia control group (P=0.044, 0.001). Content of ATP of cells in hypoxia combine group was obviously higher than that of cells in hypoxia KGF group and hypoxia HIF-1α group (P=0.011, 0.020). (6) After being cultured for 24 h, protein expressions of p53 of cells in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group were obviously higher than that of cells in normoxia control group (P<0.001), and protein expression of p53 of cells in hypoxia combine group was obviously lower than those of cells in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group (P=0.001, 0.001, 0.002). Conclusions KGF combined with HIF-1α have significant protective effects on IEC-6 of rats with hypoxia stress, and can improve its survival in hypoxic environment by inhibiting cell cycle arrest, reducing the level of apoptosis, and increasing level of energy metabolism.
2019, 35(1): 61-61. doi: 10.3760/cma.j.issn.1009-2587.2019.01.101
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Brief Original Article
Effects of free superficial temporal fascia flaps and skin grafts in repairing deep wounds in posterior ankle region of extensively burned patients
Yang Xuekang, Chen Qiaohua, Zhang Yue, Li Zhiqiang, Tao Ke, Han Juntao, Hu Dahai
2019, 35(1): 62-64. doi: 10.3760/cma.j.issn.1009-2587.2019.01.011
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Objective To observe the effects of the method of combining free superficial temporal fascia flaps with skin grafts in repairing deep wounds in posterior ankle region of extensively burned patients. Methods From September 2013 to February 2017, 11 extensively burned patients with deep tissue defects in posterior ankle region were treated in our unit. Two patients had tissue defects in bilateral posterior ankle regions. The wound sizes ranged from 5.8 cm×4.6 cm to 11.7 cm×5.2 cm. Free superficial temporal fascia flaps with the same sizes as the wounds were designed and resected to repair wounds in posterior ankle regions after debridement. The proximal end of superficial temporal veins and posterior tibial veins were performed with end-to-end anastomosis, and superficial temporal arteries and posterior tibial arteries were performed with end-to-side anastomosis. Skin grafts were resected to cover the superficial temporal fascia flaps according to patients′ condition of donor sites. The donor sites were sutured directly. Results All fascial flaps in 11 patients survived, while 2 skin grafts had partial necrosis, and they healed after secondary skin graft. All patients were followed up for 6 to 13 months, and the shape and function of the operation sites in all patients recovered well. Conclusions The method of combining free superficial temporal fascia flaps with skin grafts can repair deep wounds in posterior ankle regions of extensively burned patients. It has the advantages of good appearances in the recipient sites, strong resistances to infection of fascia flaps, minimal damages to the donor sites, short course of disease, and good prognosis of patients.
Clinical effects of perforating branch flaps of medial vastus muscle in repairing secondary wounds in donor sites of free anterolateral femoral perforator flaps
Gao Qiufang, Zhang Xiaofeng, Zhang Wanfeng, Ma Bin, Niu Xuetao, Ma Yajun, Li Zibiao, Liu Ning
2019, 35(1): 65-68. doi: 10.3760/cma.j.issn.1009-2587.2019.01.012
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Objective To investigate the clinical effects of perforating branch flaps of medial vastus muscle in repairing secondary wounds in donor sites of free anterolateral femoral perforator flaps. Methods From August 2014 to December 2016, 12 patients (8 males and 4 females, aged 35-72 years) with skin and soft tissue defects of extremities associated with tendon and bone exposure were treated in Hanzhong Central Hospital. The sizes of the primary wounds after debridement were 10 cm×8 cm-22 cm×14 cm, and the wounds were repaired with 12 cm×10 cm-24 cm×16 cm free anterolateral femoral perforator flaps. The anterolateral femoral donor sites, which were 8.0 cm×4.0 cm-14.0 cm×7.5 cm in the secondary wounds after skin extensional suture, were repaired with perforating branch flaps of medial vastus muscle in the size of 9.0 cm×5.0 cm-15.0 cm×8.5 cm. The medial femoral donor sites were sutured directly. Results All the perforating branch flaps of medial vastus muscle and free anterolateral femoral perforator flaps survived in 12 patients. Following up for 6 to 12 months, the medial femoral perforator flaps had good local shape and texture. The flaps of 8 patients without cutaneous nerve transection were sensitive. The sensation of the flaps of the other 4 patients gradually recovered, and the functions of the ipsilateral knee joints were normal. Conclusions The medial femoral perforator flap has a stable anatomy and abundant blood supply, which can be used to repair the secondary wound in the donor site of the free anterolateral femoral perforator flap conveniently. It is safe and easy to be popularized. Moreover, it has a good shape and function after operation.
Case Report
One case of pyoderma gangrenosum in the lower limbs in Tibetan Plateau treated with vacuum sealing drainage combined with irrigation of oxygen loaded fluid
Yang Sisi, Jiang Shuguo, Tudeng Cicheng, Ni zhen, Xiao Chengzhi
2019, 35(1): 69-71. doi: 10.3760/cma.j.issn.1009-2587.2019.01.013
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A 54 years old male patient with chronic leg ulcers was admitted in our hospital in November 2017. He was diagnosed as pyoderma gangrenosum by the pathological examination. Then the wound was treated with simple vacuum sealing drainage combined with irrigation of oxygen loaded fluid. This therapy overcame the shortage of hypoxia in the Tibetan Plateau on wound healing, resulting in a better wound healing. The patient was eventually cured and discharged from hospital.
Acquired blood coagulation factor Ⅴ deficiency in a patient with severe burn
Bi Xiaojie, Jin Xianfu, Zhang Huifei, Su Zhengxian, Shen Bo
2019, 35(1): 71-73. doi: 10.3760/cma.j.issn.1009-2587.2019.01.014
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In March 2017, a severely burned male patient aged 36 years with hypovolemic shock was admitted to our hospital. The patient received large quantities of antibiotics and blood products and repeated skin graft after admission, and then he suffered wound errhysis and throat congestion. The patient was healthy before without family history of bleeding or thrombosis disease. Laboratory tests showed that prothrombin time and activated partial coagulation time were remarkably prolonged, blood coagulation factor Ⅴ activity was extremely low, and the result of qualitative test of coagulation factor inhibitor was positive. Acquired blood coagulation factor Ⅴ deficiency was diagnosed. After application of dexamethasone (5 mg, twice per day) and infusion of fresh frozen plasma, blood coagulation indicators of patients recovered in 4 days, the result of qualitative test of coagulation factor inhibitor was negative, and bleeding symptoms were improved.
Review
Advances in the research of transcatheter arterial thrombolysis for severe frostbite therapy
Zhang Jinlong, Fu Jinxin, Yuan Kai, Yuan Bing, Wang Maoqiang
2019, 35(1): 74-76. doi: 10.3760/cma.j.issn.1009-2587.2019.01.015
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Severe frostbite (grade Ⅲ to Ⅳ) is a common disease accompanied with high disability rate in cold regions, especially for military training and disaster events in cold regions. The treatment of severe frostbite mainly includes rapid rewarming in the early stage and amputation in the later stage; while the damage of vascular endothelial cells, microvascular thrombosis, and decreased tissue perfusion secondary to severe frostbite are important factors affecting prognosis. Transcatheter arterial thrombolysis is a new technique for the treatment of severe frostbite. It has the advantages of minimally invasive, high safety, and significantly reduced amputation rate. We reviewed the advances in the research of transcatheter arterial thrombolysis for the treatment of severe frostbite.
Advances in exosomes derived from the mesenchymal stem cells in the treatment of sepsis
Wang Shan, Yin Haiyan
2019, 35(1): 77-80. doi: 10.3760/cma.j.issn.1009-2587.2019.01.016
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Exosomes derived from mesenchymal stem cells (MSCs) are small membrane vesicles with diameters in 30-150 nm, which are secreted into extracellular matrix by MSCs in resting or activated state. Recent studies have found that exosomes secreted by MSCs can be used as important signal transduction mediator, which can transport bioactive substances such as mRNA, microRNA, and proteins effectively to target cells, and play important roles in regulating tissue regeneration and immunomodulation. Here we give an overview of biological properties, mechanisms, and the roles in treating sepsis of exosome derived from MSCs, in order to provide some basises to highlight the MSCs-exosomes therapy in sepsis.

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