中华烧伤与创面修复杂志

) thermo-sensitive hydrogel on the wound healing of full-thickness skin defects in diabetic mice. Methods (1) According to the volume ratio of bacteria to medium of 1∶100, about 5×10⁸ colony forming units/mL (the same concentration below)

L

.

lactis

was cultured in M17GS liquid medium. The growth conditions were observed at 0 (immediately), 2, 4, 6, 8, 10, and 12 h of culture with a microplate reader. In addition, another colony of the bacteria was taken and cultured under the same condition mentioned above. The culture medium was collected at the same time points as mentioned above, and the supernatant of bacterial culture was isolated. With the supernatant, the pH value was measured with a desktop pH meter, and the concentration of L-lactic acid at 0 (immediately), 2, 4, 8, and 12 h of culture was determined by the L-lactic acid detection and analysis kit (

n

=3). (2) To prepare a simple thermo-sensitive hydrogel, the poloxamer thermo-sensitive polymer and M17GS liquid medium were mixed thoroughly according to the mass-volume ratio of 0.2 g∶1 mL.

L

.

lactis

was added to the simple thermo-sensitive hydrogel according to the volume ratio of bacteria to hydrogel of 1∶100, and the

L

.

lactis

thermo-sensitive hydrogel was prepared after thorough mixing. Afterwards, the morphology of

L

.

lactis

thermo-sensitive hydrogel was observed after 4 ℃, 37 ℃ incubation and again at 4 ℃ incubation after gelation. The storage modulus and loss modulus of the

L

.

lactis

thermo-sensitive hydrogel at 10-40 ℃ were measured by rheometer, and the gel forming temperature was observed. After freeze-drying the

L

.

lactis

thermo-sensitive hydrogel, the surface and the morphological structure of

L

.

lactis

in the hydrogel were observed by scanning electron microscope. (3) Mouse macrophages Raw264.7 cells were M1-type polarization stimulated by culturing with lipopolysaccharide and interferon γ in the final mass concentration of 100 and 10 ng/mL respectively for 24 h. The cells were divided into blank control group (without other treatment),

L

.

lactis

thermo-sensitive hydrogel group, and lactic acid group.

L

.

lactis

thermo-sensitive hydrogel in the volume of 1 mL was added to the cells of

L

.

lactis

thermo-sensitive hydrogel group, while lactic acid with the final molarity of 30 mmol/L was added to the cells in lactic acid group. After being cultured at 37 ℃ for 24 h, mRNA expressions of the markers arginase 1 and CD206 of M2-type macrophages were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) (

n

=3), and the immunofluorescence method was used to detect the protein localization and expression of arginase 1 and CD206. (4) Fifteen female BALB/c mice aged 8-10 weeks were induced into diabetic mouse models by the method of streptozotocin combined with high-sugar and high-fat diet, and a full-thickness wound with the diameter of 6 mm was made on the back of each mouse. The mice were divided into blank control group (without other treatment), thermo-sensitive hydrogel alone group, and

L

.

lactis

thermo-sensitive hydrogel group according to the random number table, with 5 mice in each group. The mice in the hydrogel treatment two groups were dripped with 200 μL corresponding hydrogel to the wound surface immediately after injury, and the hydrogel was replaced every day. After treatment for 0 (immediately), 3, 6, 9, and 12 days in the hydrogel treatment two groups, wound healing was observed, and wound area was measured. After 12 days of treatment, the wound tissue was taken to observe the thickness of granulation tissue by hematoxylin-eosin staining and CD206 and the marker of M1-type macrophages of inducible nitric oxide synthase (iNOS) positive cells by immunofluorescence method. The mice in blank control group were observed at the same time points as mentioned above. (5) Nine female BALB/c mice aged 8-10 weeks were induced into diabetic mouse models by the same method of experiment (4). Then, they were divided into normal skin group (without other treatment), wound alone group, and

L

.

lactis

thermo-sensitive hydrogel group according to the random number table, with 3 mice in each group. Mice in wound alone group and

L

.

lactis

thermo-sensitive hydrogel group were prepared with full-thickness skin defect wounds according to the method of experiment (4). Mice in the former group was left untreated after injury, and in the latter group, 200 μL

L

.

lactis

thermo-sensitive hydrogel was dripped onto the wound surface immediately after injury. After treatment for 1 day in hydrogel treatment group, the wound tissue of mice was taken, and the mRNA expressions of interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), and nuclear factor κB were detected by real-time fluorescence quantitative RT-PCR; after the eyeball blood was collected, the leukocyte count, lymphocyte count, and monocyte count in peripheral blood were measured by an automatic blood cell analyzer, and the serum L-lactic acid concentration was measured by the L-lactic acid detection and analysis kit. At the same time point mentioned above, normal skin tissue was taken from the corresponding parts of mice in normal skin group, wound tissue was taken from mice in wound alone group, and blood was taken from mice of the two groups for corresponding detection. Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, Tukey, and Dunnett test. Results (1) The growth of

L

.

lactis

reached the plateau in about 6 h of culture. In the culture supernatant of

L

.

lactis,

the pH value gradually decreased, reaching the nadir about 4.9 after 8 h of culture, and the L-lactic acid concentration gradually increased, which peaked about 70 mmol/L after 8 h of culture. (2) The

L

.

lactis

thermo-sensitive hydrogel was a liquid at 4 ℃, and a solid gel at 37 ℃. After gelation, it became a liquid again after incubating at 4 ℃. The gel forming temperature was about 25 ℃. The storage modulus was about 3 000 Pa, and the loss modulus was about 1 000 Pa after gelation. Under the scanning electron microscope, the

L

.

lactis

thermo-sensitive hydrogel showed a loose three-dimensional porous structure, and the

L

.

lactis

had an ellipsoidal shape being wrapped inside the hydrogel. (3) After 24 h of culture, compared with those in blank control group, the expression of arginase 1 increased significantly (

q

=11.620, 15.250,

P

<0.01), the expression of CD206 mRNA increased significantly (

q

=16.770, 19.030,

P

<0.01), and the expression of CD206 protein located in the cell membrane and arginase 1 protein located in the cytoplasm increased significantly in the macrophages of

L

.

lactis

thermo-sensitive hydrogel group and lactic acid group. The expressions of arginase 1 and CD206 mRNA in the macrophages between lactic acid group and

L

.

lactis

thermo-sensitive hydrogel group were similar (

q

=3.629, 2.259,

P

>0.05). (4) After 3-12 days of treatment, compared with those in blank control group and thermo-sensitive hydrogel alone group, the wound of mice in

L

.

lactis

thermo-sensitive hydrogel group healed faster, the wound area was significantly reduced, and the inflammation of the wound edge tissue was reduced. After treatment of 3, 6, 9, 12 days, the wound areas of mice in

L

.

lactis

thermo-sensitive hydrogel group were (25.8±5.9), (21.2±4.6), (16.0±2.4), (8.4±2.4) mm² respectively, which were significantly smaller than (31.8±5.3), (28.0±3.4), (22.6±3.7), (17.0±1.0) mm² in blank control group (

q

=3.506, 3.973, 3.856, 5.025,

P

<0.05 or

P

<0.01). After treatment of 3 and 6 days, the wound areas of mice in

L

.

lactis

thermo-sensitive hydrogel group were significantly smaller than those in thermo-sensitive hydrogel alone group (

q

=3.739, 3.739,

P

<0.05). After 12 days of treatment, compared with those in blank control group and thermo-sensitive hydrogel alone group, the wound granulation tissue of mice in

L

.

lactis

thermo-sensitive hydrogel group was thicker, with significantly reduced iNOS positive cells and increased CD206 positive cells in wound tissue. (5) After 1 day of treatment, the mRNA expressions of IL-1β, TNF-α, and nuclear factor κB in the wound tissue of mice in wound alone group were significantly higher than those of normal skin tissue of mice in normal skin group (

q

=9.253, 4.819, 6.020,

P

<0.01) but similar to those in

L

.

lactis

thermo-sensitive hydrogel group (

q

=2.850, 2.735, 2.556,

P

>0.05). The peripheral blood leukocyte count, lymphocyte count, and monocyte count of mice in wound alone group were significantly higher than those in normal skin group (

q

=3.523, 5.373, 5.279,

P

<0.05 or

P

<0.01) but similar to those in

L

.

lactis

thermo-sensitive hydrogel group (

q

=0.621, 1.240, 1.293,

P

>0.05). The serum L-lactic acid concentration of mice in the three groups remained within the normal range and the overall comparison among them was not statistically significant (

F

=4.095,

P

>0.05). Conclusions The

L

.

lactis

thermo-sensitive hydrogel was safe to use locally on the wounds of diabetic mice with full-thickness skin defects. It can produce and deliver lactic acid in situ, promote the polarization of macrophages from M1 to M2, reshape the wound healing microenvironment, and promote efficient wound healing.

Influence of porcine urinary bladder matrix and porcine acellular dermal matrix on wound healing of full-thickness skin defect in diabetic mice

Zhao Peng, Yang Minlie, Chu Guoping, Jia Zhigang, Zhou Xiaojin, Lyu Guozhong

2020, 36(12): 1130-1138. doi: 10.3760/cma.j.cn501120-20200901-00399

Abstract

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Abstract:

Objective To compare the difference of pro-healing effect of porcine urinary bladder matrix (UBM) and porcine acellular dermal matrix (ADM) on full-thickness skin defect wounds in diabetic mice. Methods Thirty-six type 2 diabetic BKS db/db mice aged 10 weeks were divided into UBM group and ADM group according to the random number table, with 18 mice in each group and preoperative molarity of non-fasting blood glucose higher than 16.6 mmol/L. A circular full-thickness skin defect wound with 6 mm in diameter was made on the back of each mouse, and porcine UBM and porcine ADM scaffolds were implanted into the wounds of both groups correspondingly. Immediately after operation and on post operation day (POD) 7, 14, and 28, wounds were observed generally. On POD 7, 14, and 28, 6 mice of each group were collected respectively to calculate the rate of wound epithelialization, and then the corresponding mice were sacrificed after calculation, and the wound tissue was harvested to make slices. Six slices of the mice in the 2 groups on POD 7 and 14 were respectively collected to stain with haematoxylin-eosin (HE), and 6 slices on POD 7 and 28 had Masson′s staining, which were used to observe histopathological changes and scaffold degradation. On POD 7 and 14, 24 slices of each mouse in the 2 groups were collected respectively to detect alpha smooth muscle actin (α-SMA) and CD31 positive expression denoting the growth of myofibroblasts and neovessels respectively and observe the distribution and activation of macrophages with immunohistochemical staining. The wound tissue of mice in the 2 groups on POD 7 and 14 was harvested to detect mRNA expressions of fibroblast growth factor 2 (FGF-2), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and transforming growth factor β₁ (TGF-β₁) by real-time fluorescence quantitative reverse transcription polymerase chain reaction. The sample number of above-mentioned indexes in each group at each time point was 6. Data were statistically analyzed with analysis of variance for factorial design,

t

test, and Bonferroni correction. Results (1) General observation showed that integration of UBM scaffold into the wounds of mice in UBM group on most time points was superior, and integration of ADM scaffold into the wounds of mice in ADM group on most time points was inferior. On POD 28, epidermis still did not form in some region of scaffold surface of wounds of mice in ADM group, while wounds of mice in UBM group were completely epithelialized. On POD 7, 14, and 28, wound epithelialization rates of mice in UBM group were respectively (22.4±6.4)%, (68.6±12.4)%, and 100.0%, all significantly higher than (4.5±2.2)%, (23.6±4.6)%, and (64.2±13.2)% in ADM group (

t

=7.427, 9.665, 7.655,

P

<0.01). (2) HE staining and Masson′s staining showed that a large number of cells appeared in wound scaffold of mice in UBM group on POD 7; cells distributed in the whole region of UBM scaffold on POD 14; dermal tissue with structure similar to normal skin formed in the wounds and the fibrous morph of UBM scaffolds disappeared on POD 28. Only a small number of cells appeared in inside of wound scaffolds of mice in ADM group on POD 7; on POD 14, cells were sparsely distributed in ADM scaffolds; on POD 28, the morph of originally robust collagen fiber of ADM scaffolds was still clear and visible. (3) On POD 7, a large number of accumulated myofibroblasts and neovessels appeared in the lower layers of scaffolds of wounds of mice in UBM group; on POD 14, evenly distributed myofibroblasts and neovessels appeared in the upper layers of UBM scaffolds, and most vessels were perfused. On POD 7 and 14, myofibroblasts were sparsely distributed in scaffolds of wounds of mice in ADM group with no or a few neovascular structures perfused unobviously. On POD 7 and 14, α-SMA positive expressions in scaffolds of wounds of mice in UBM group were significantly higher than those in ADM group (

t

=25.340, 6.651,

P

<0.01); CD31 positive expressions were also significantly higher than those in ADM group (

t

=34.225, 10.581,

P

<0.01). (4) On POD 7, a large number of macrophages appeared in the lower layers of scaffolds of wounds of mice in UBM group; on POD 14, macrophages infiltrated into the internal region of UBM scaffolds, and M2 polarization occured without M1 polarization. On POD 7, a small number of macrophages appeared in the bottom of scaffolds of wounds of mice in ADM group; on POD 14, macrophages were few in internal region of ADM scaffold, and neither M2 polarization nor M1 polarization occurred. (5) On POD 7 and 14, mRNA expressions of FGF-2, VEGF, PDGF, and TGF-β₁ in the wound tissue of mice in UBM group were all significantly higher than those in ADM group (

t

=7.007, 14.770, 10.670, 8.939; 7.174, 7.770, 4.374, 4.501,

P

<0.01). Conclusions Porcine UBM scaffold is better than porcine ADM in facilitating wound repair and dermis reconstruction of full-thickness skin defects in diabetic mice through the induction of myofibroblasts and macrophages immigration, the promotion of neovascularization and expression of pro-healing growth factors.

Effects and mechanism of copper oxide nanozymes on wound healing of full-thickness skin defects in diabetic mice

Peng Yuan, Lu Yifei, Deng Jun, Zhang Yan

2020, 36(12): 1139-1148. doi: 10.3760/cma.j.cn501120-20200929-00426

Abstract

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Abstract:
Objective To investigate the effects and mechanism of copper oxide nanozymes on wound healing of full-thickness skin defects in diabetic mice. Methods (1) Copper oxide nanozymes were synthesized through the reaction of copper chloride and L-ascorbic acid. Transmission electron microscope was used for observing the particle size and morphology of copper oxide nanozymes, and dynamic light scattering particle size analyzers and Zeta potentiometer were used to analyze the hydrated particle size and surface potential of copper oxide nanozymes, respectively. (2) The hydrogen peroxide detection kit, superoxide anion determination kit, and 3, 3′, 5, 5′-tetramethylbenzidine were used to test the hydrogen peroxide, superoxide anion, and hydroxyl radicals scavenging ability of 150 ng/mL copper oxide nanozymes, respectively, and the scavenging proportions of hydrogen peroxide, superoxide anion, and hydroxyl radicals were calculated. The sample numbers were all 3. (3) Mouse fibroblast cell line 3T3 cells were divided into blank control group, simple hydrogen peroxide group, and hydrogen peroxide+ copper oxide group according to the random number table (the same grouping method below), with 3 wells in each group. Cells in hydrogen peroxide+ copper oxide group were pre-treated with copper oxide nanozymes in final mass concentration of 25 ng/mL for 30 minutes, and then hydrogen peroxide in final molarity of 250 μmol/L was added into simple hydrogen peroxide group and hydrogen peroxide+ copper oxide group. Cells in blank control group were routinely cultured. After 24 hours of culture, 2′, 7′-dichlorodihydrofluorescein diacetate fluorescence probe was used to detect the level of reactive oxygen species (indicated by green fluorescence intensity) in cells and cell counting kit-8 assay was performed to detect and calculate the cell survival rate. (4) Ten male BALB/c mice aged 6-8 weeks (the same gender and age below) were divided into phosphate buffer saline (PBS) group and copper oxide group, with 5 mice in each group. The mice in the copper oxide group were injected with 800 ng/kg copper oxide nanozyme at a concentration of 200 ng/mL via the caudal vein, and the mice in PBS group were treated with the same volume of PBS. The mice in the two groups were treated once a day for seven consecutive days. On the eighth day, 5 mice from each group were conducted and blood samples were taken for analysis of blood panel and serum biochemistry, and then the heart, liver, spleen, lung, and kidney were harvested for histopathological observation by hematoxylin-eosin (HE) staining after the mice were sacrificed. (5) Twenty mice were divided into PBS group and copper oxide group, with 10 mice in each group. Diabetes was induced by streptozotocin and high-sugar and high-fat diet and a full-thickness skin defect wound with diameter of 6 mm was reproduced on the back of each diabetic mouse. Immediately after injury, 20 μL PBS and 20 μL copper oxide nanozymes at the concentration of 200 ng/mL were added respectively to the wounds of mice in PBS group and copper oxide group, with the treatment being continued for twelve consecutive days. Three mice were selected from each group, and the wound healing was observed on post injury day (PID) 0 (immediately), 3, 6, 9, and 12 and the un-healed area was calculated. On PID 6, three mice from each group that were not for wound observation were sacrificed, and the content of interleukin 1β (IL-1β), tumor necrosis factor-α (TNF-α), and IL-6 in the wound tissue were determined by enzyme-linked immunosorbent assay. On PID 12, the rest 7 mice in each group were sacrificed for observation of the length of regenerated epidermis in wound tissue by HE staining, and the level of reactive oxygen species (indicated as red fluorescence intensity) in wound tissue by dihydroethidium staining. Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, independent sample t test, and Bonferroni test. Results (1) The prepared copper oxide nanozymes were uniform in size with an average diameter of 3.5-4.0 nm in dry state, the hydrated particle size of 4.5 nm, and the surface potential of (-9.8±0.3) mV. By comprehensive judgment, copper oxide nanozymes had been successfully prepared. (2) After being treated with copper oxide nanozyme for 2 hours, 10 minutes, and 5 minutes, respectively, the scavenging proportions of hydrogen peroxide, superoxide anion, and hydroxyl radicals were (77±5)%, (45±5)%, and (84±4)%, respectively. (3) After 24 hours of culture, the cells in simple hydrogen peroxide group showed a significantly increased level of reactive oxygen species with abnormal morphology and decrease in cell number, while the cells in hydrogen peroxide+ copper oxide group showed a remarkably decreased level of reactive oxygen species with normal morphology similar to that of blank control group. The cell survival rate in simple hydrogen peroxide group was obviously reduced compared with the rates in blank control group and hydrogen peroxide+ copper oxide group (P<0.01), while there was no significant difference in cell survival rate between hydrogen peroxide+ copper oxide group and blank control group. (4) After 7 days of injection, there were no obvious differences in liver and kidney function indexes and blood panel indexes between mice in PBS group and copper oxide group. No necrosis, hyperaemia or hemorrhage in heart, liver, spleen, lung, or kidney was observed in mice in copper oxide group, which was similar to that in PBS group. (5) Compared with that of PBS group, wounds of mice in copper oxide group showed an accelerated healing trend with less redness. On PID 6, 9, and 12, the areas of un-healed wound of mice in copper oxide group (28.8±1.9), (17.6±3.8), and (10.4±1.8) mm², respectively, significantly lower than (38.0±4.3), (30.2±3.0), and (24.2±3.0) mm² in PBS group (t=3.706, 5.075, 5.558, P<0.01). On PID 6, the content of IL-1β, TNF-α, and IL-6 in wounds of mice in copper oxide group were significantly lower than that in PBS group (t=6.115, 11.762, 11.725, P<0.01). On PID 12, the length of regenerated epidermis in wounds of mice in copper oxide group was obviously longer than that in PBS group, the level of reactive oxygen species in wounds of mice in copper oxide group was obviously lower than that in PBS group. Conclusions Copper oxide nanozyme not only has good biocompatibility, but also has efficient reactive oxygen species scavenging activity. It can eliminate the over-expressed reactive oxygen species in the full-thickness defect wounds of diabetic mice, reduce oxidative stress and inflammation, thus promoting wound repair.

Clinical efficacy and influencing factors of different modes of continuous negative pressure wound therapy on venous ulcer wounds of lower limbs

Yang Minlie, Zhou Xiaojin, Zhu Yugang, Jiang Donglin, Ding Lintao, Chu Guoping, Zhao Peng, Cheng Jia, Lyu Guozhong, Li Qingfeng

2020, 36(12): 1149-1158. doi: 10.3760/cma.j.cn501120-20200316-00173

Abstract

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Abstract:
Objective To explore the clinical efficacy of different modes of continuous negative pressure wound therapy (NPWT) on venous ulcer wounds of lower limbs, and to analyze the influencing factors. Methods From January 2018 to December 2019, 53 patients with venous ulcer of lower limbs who met the inclusion criteria and hospitalized in the Affiliated Hospital of Jiangnan University were recruited in this prospective randomized controlled study. According to the random number table, the patients were divided into single negative pressure therapy (SNPT) group (19 patients, 11 males and 8 females), cyclic alternating negative pressure therapy (CANPT) group (17 patients, 12 males and 5 females), and routine dressing change (RDC) group (17 patients, 10 males and 7 females), aged (47±11), (49±10), and (47±10) years respectively. After admission, patients in SNPT group were given continuous NPWT with the single negative pressure setting at -13.3 kPa, patients in CANPT group were also given continuous NPWT but with the cyclic alternating negative pressure setting from -16.0 to -10.7 kPa, while patients in RDC group were given dressing change with vaseline gauze soaked with iodophor. The wound healing rate was calculated on treatment day 7 and 14. Transcutaneous oxygen pressure (TcPO₂) around the wound was detected by TcPO₂ meter before treatment and on treatment day 7 and 14. The wound exudate/drainage fluid was collected on treatment day 1, 4, 7, 10, and 14, with the pH value measured using a pH meter, and the volume of exudate/drainage fluid recorded. Before treatment and on treatment day 7 and 14, venous blood was collected to detect the serum levels of interleukin 1β (IL-1β), IL-6, tumor necrosis factor α(TNF-α), transforming growth factor-β₁ (TGF-β₁), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Before treatment and on treatment day 7 and 14, wound exudates were collected for bacterial culture, and Visual Analogue Scale and Hamilton Anxiety Scale were used to evaluate the degree of wound pain and anxiety of patients respectively. The length of hospital stay and the total treatment cost were counted. Analysis of variance for repeated measurement, one-way analysis of variance, least significant difference test, Kruskal Wallis H test, Mann Whitney U test, chi-square test, Fisher′s exact probability method test, and Bonferroni correction were used to analyze the data. According to the wound healing rate on treatment day 14, the efficiency of patients were divided into two grades of significant healing with wound healing rate≥70% and non significant healing with wound healing rate<70%. According to the two categories of wound healing rate as dependent variables, the levels of TcPO₂, IL-1β, IL-6, TNF-α, TGF-β₁, VEGF, bFGF levels and bacterial detection, wound pain and anxiety before treatment, wound exudate/drainage fluid volume and pH value on treatment day 1 were taken as covariates, and binary classification multifactor logistic regression analysis was used to analyze the risk factors of significant wound healing. Results (1) On treatment day 7, the wound healing rate of patients in SNPT group was (33±10) %, which was significantly higher than (24±9) % of RDC group (P<0.05). On treatment day 14, the wound healing rates of patients in SNPT group and CANPT group were (71±15)% and (66±18)%, respectively, which were significantly higher than (45±19)% of RDC group (P<0.01). (2) Compared with those of RDC group, the TcPO₂ value around the wound of patients was significantly increased in SNPT group on treatment day 14 and in CANPT group on treatment day 7 and 14 (P<0.05 or P<0.01), the pH value of wound drainage fluid of patients was significantly decreased in SNPT group on treatment day 10 and 14 and in CANPT group on treatment day 7 and 14 (P<0.05), the volume of wound drainage fluid of patients was significantly reduced in SNPT group on treatment day 10 and 14 and in CANPT group on treatment day 7, 10, and 14 (Z=-4.060, -4.954, -2.413, -4.085, -4.756, P<0.05 or P<0.01), the serum levels of IL-1β, IL-6, and TNF-α of patients were significantly decreased in SNPT group and CANPT group on treatment day 7 and 14 (P<0.01), the serum level of TGF-β₁ of patients was significantly increased in CANPT group on treatment day 14 (P<0.05), the serum levels of VEGF and bFGF were significantly increased in SNPT group and CANPT group on treatment day 14 (P<0.01), the bacteria detection proportion of wound exudate, wound pain, and anxiety scores of patients were significantly decreased in SNPT group and CANPT group on treatment day 7 and 14 (P<0.01). Compared between the negative pressure therapy two groups, except the wound pain score of patients in CANPT group was significantly lower than that in SNPT group (P<0.01) on treatment day 7, the other indicators mentioned above were similar. (3) The length of hospital stay of patients in SNPT group was similar to that in CANPT group (P>0.05), which were significantly shorter than the time in RDC group (P<0.01). The total treatment cost of patients among the three groups was similar (F=1.766, P>0.05). (4) Before treatment, the serum levels of TNF-α and bFGF, TcPO₂ around the wound, and the degree of wound pain were risk factors for significant wound healing (odds ratio=1.109, 0.950, 1.140, 2.169, 95% confidence interval=1.012-1.217, 0.912-0.988, 1.008-1.290, 1.288-3.651, P<0.05 or P<0.01). Conclusions Clinical application of continuous NPWT under single negative pressure mode and cyclic alternating negative pressure mode has a positive effect on improving the wound base and healing rate of venous ulcer of lower limbs. But cyclic alternating negative pressure mode is significantly more effective than single negative pressure mode in improving TcPO₂ around the wound, reducing wound pH value, reducing exudate volume and relieving pain. The serum levels of TNF-α and bFGF, TcPO₂ around the wound and the degree of wound pain were the risk factors that affect the wound healing significantly.

Analysis of the dynamic changes in gut microbiota in patients with extremely severe burns by 16S ribosomal RNA high-throughput sequencing technology

Pan Yanyan, Fan Youfen, Li Jiliang, Cui Shengyong, Huang Neng, Jin Guoying, Chen Cui, Zhang Chun

2020, 36(12): 1159-1166. doi: 10.3760/cma.j.cn501120-20200518-00271

Abstract

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Abstract:
Objective To analyze the dynamic change rule of gut microbiota in patients with extremely severe burns using 16S ribosomal RNA (rRNA) high-throughput sequencing technology. Methods Five patients with extremely severe burns who were admitted to Hwa Mei Hospital of University of Chinese Academy of Sciences from February to June 2017 and conformed to the inclusion criteria were included in the prospective observational study. All patients were males with age of 32-48 years. Fecal samples were collected in the shock stage (within 3 days after injury), early stage of acute infection (4-14 d after injury), middle stage of acute infection (15-28 d after injury), late stage of acute infection (from 29 d after injury to 1 week before discharge) and within 1 week before discharge. The number of samples was 5 in each stage. The fecal pH value was measured using a pH meter. High-throughput sequencing technology was applied for sequencing of 16S rRNA V3 and V4 regions of fecal samples. QIIME software was used to analyze the number of operational taxonomic units (OTUs), α diversity (Chao1 index and Shannon index), and the relative abundance of gut microbiota at the phylum and family levels. Unweighted pair group method with arithmetic mean clustering method was used to analyze the β diversity of gut microbiota, and Tax4Fun was used to predict functional changes of gut microbiota. Data were statistically analyzed with one-way analysis of variance for repeated measurement, Bonferroni method, Wilcoxon rank sum test for paired samples, and Bonferroni correction. Results (1) The pH value of feces in the early and middle stages of acute infection in patients with extremely severe burns in this group was 7.40±0.45 and 7.56±0.45 respectively, which were significantly higher than 6.68±0.36 in the shock stage (P<0.05 or P<0.01). (2) A total of 2 333 584 efficient and high-quality sequences were obtained, and the length of the sequences was about 415 bp. A total of 1 209 OTUs were obtained. The sequencing coverage of all samples was over 99.0%. The number of OTUs and Chao1 index in the early, middle, and late stages of acute infection in patients with extremely severe burns in this group were significantly lower than those in the shock stage (Z=2.023, P<0.05). The number of OTUs and Chao1 index within 1 week before discharge were significantly higher than those in the early, middle, and late stages of acute infection, and Shannon index within 1 week before discharge was significantly higher than that in the early and middle stages of acute infection (Z=2.023, P<0.05). (3) The structure of gut microbiota in the shock stage in patients with extremely severe burns in this group was highly similar to that within 1 week before discharge, and lowly similar to that in the early, middle, and late stages of acute infection. The analysis of individual sample showed that the clustering rule of most of the samples was in accordance with that of the staged samples. The weighted Unifrac distance of gut microbiota in the shock stage was significantly shorter than that in the early, middle, and late stages of acute infection (Z=3.326, 2.570, 2.690, P<0.05 or P<0.01), while the weighted Unifrac distance of gut microbiota in the other stages was similar. (4) At the phylum level, compared with that in the shock stage, the relative abundance of Firmicutes was decreased in the early, middle, and late stages of acute infection, while the relative abundance of Bacteroidetes and Proteobacteria increased. However, the relative abundance of the above three phyla within 1 week before discharge was similar to that in the shock stage. At the family level, the top five dominant bacteria in relative abundance in different stages after injury were quite different. The relative abundance of dominant five family bacteria in the shock stage was decreased in the early, middle, and late stages of acute infection. The relative abundance of non-dominant bacteria such as Enterobacteriaceae, Streptococcaceae, and Bacteroidaceae in the shock stage increased significantly in the early, middle, and late stages of acute infection, which became new dominant families in these stages. The relative abundance of some acid-producing bacteria within 1 week before discharge resumed to the similar level in the shock stage. (5) Functions such as some amino acid metabolism, glycolysis and gluconeogenesis, and pyruvate metabolism of gut microbiota were obviously weaker in the early and middle stages of acute infection than those in the shock stage. Functions such as some amino acid metabolism and carbohydrate metabolism of gut microbiota were significantly enhanced in the late stage of acute infection compared with that in the shock stage. The distributions of functional genes in gut microbiota were similar between the shock stage and within 1 week before discharge. Conclusions The internal environment and gut microbial compositions in extremely severe burned patients change significantly in the early and middle stages of acute infection. The pH value increases, the bacterial species and diversity decrease, especially the relative abundance of acid-produced bacteria is significantly reduced, which gradually recover with the improvement of the patient′s condition. The pH value and the changes of Proteobacteria and acid-producing bacteria could be considered as suitable parameters for reflecting the disorder level of gut microbiota in patients with extremely severe burns.

Value of blood routine indexes and their ratios in judging the prognosis of adult patients with extensive burns

Zheng Jianjun, Wang Zi′en, Zheng Linwen, Xu Zhaorong, Chen Shun, Chen Zhaohong

2020, 36(12): 1167-1172. doi: 10.3760/cma.j.cn501120-20200308-00133

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Objective To investigate the value of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and blood platelet count (BPC) in judging the prognosis of adult patients with extensive burns. Methods From January 2012 to December 2018, 99 adult patients with extensive burns who met the inclusion criteria were admitted to Union Hospital of Fujian Medical University, including 76 males and 23 females, aged 18 to 75 (43±13) years. According to the prognosis, the patients were divided into survival group of 79 cases and death group of 20 cases. Their clinical data were retrospectively analyzed by the method of case-control study. The gender, age, total burn area, inhalation injury, use of mechanical ventilation and white blood cell count, neutrophil count, lymphocyte count, and BPC on post injury day (PID) 1, 3, and 7 were collected, and the NLR, PLR, difference value of BPC on PID 3 and PID 1 (ΔBPC3), difference value of NLR on PID 3 and PID 1 (ΔNLR3), difference value of PLR on PID 3 and PID 1 (ΔPLR3), difference value of BPC on PID 7 and PID 1 (ΔBPC7), difference value of NLR on PID 7 and PID 1 (ΔNLR7), difference value of PLR on PID 7 and PID 1 (ΔPLR7) of patients in the two groups were calculated. Data were statistically analyzed with Mann-Whitney U test, independent sample t test, chi-square test to screen the death-related factors of patients. Binary classification single factor and multifactor logistic regression analysis were used to analyze the death-related factors of patients. The receiver′s operating characteristic (ROC) curve of the independent risk factor of death of patients predicting the prognosis of adult patients with extensive burns was drawn, and the area under the curve, the optimal threshold and its sensitivity and specificity were calculated. Results (1) There were statistically significant differences in total burn area and use of mechanical ventilation of patients between the two groups (Z=-2.615, χ²=7.282, P<0.01). (2) On PID 1, there was statistically significant difference in NLR of patients between the two groups (Z=-2.414, P<0.05). On PID 3, there were statistically significant differences in BPC and ΔNLR3 of patients between the two groups (Z=-2.048, -2.780, P<0.05 or P<0.01). On PID 7, there were statistically significant differences in lymphocyte count, BPC, NLR, and ΔNLR7 of patients between the two groups (Z=-2.248, -2.231, -2.641, -3.669, P<0.05 or P<0.01). (3) Binary classification single factor logistic regression analysis showed that the total burn area, mechanical ventilation, BPC and NLR on PID 7, and ΔNLR7 were related to death of patients (odds ratio=1.038, 0.193, 0.990, 1.086, 1.105, 95% confidence interval=1.010-1.067, 0.062-0.598, 0.982-0.998, 1.012-1.165, 1.037-1.178, P<0.05 or P<0.01). Binary classification multifactor logistic regression analysis showed that ΔNLR7 was the independent risk factor of death of adult patients with extensive burns (odds ratio=1.090, 95% confidence interval=1.008-1.178, P<0.05). (4) The optimal threshold of ROC curve of ΔNLR7 for predicting the prognostic death of 97 adult patients with extensive burns was -0.073 4. The sensitivity under the optimal threshold was 65.0%, and the specificity was 78.5%. The area under the ROC curve was 0.776 (95% confidence interval=0.650-0.882, P<0.01). Conclusions Dynamic monitoring of NLR and BPC is of great significance to assist in judging the prognosis of adult patients with extensive burns. ΔNLR7 is an independent predictor of death in adult patients with extensive burns, while PLR can not predict the death of adult patients with extensive burns.

Effects and mechanism of norepinephrine on the migration of bone marrow mesenchymal stem cells in mice

Kong Yanan, Jin Jing, Cheng Biao

2020, 36(12): 1173-1182. doi: 10.3760/cma.j.cn501120-20200325-00194

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Objective To investigate the effects and mechanism of sympathetic neurotransmitter norepinephrine (NE) on the migration of bone marrow mesenchymal stem cells (BMSCs) in mice. Methods (1) Twenty 3-week-old male C57BL/6 mice were sacrificed for isolating, culturing, and identifying BMSCs from the femur and tibia. Cells of the second or third passages were divided into phosphate buffer solution (PBS) group, 1 μmol/L NE group, 10 μmol/L NE group, and 100 μmol/L NE group, with 8 wells in each group. Cells in 1 μmol/L NE group, 10 μmol/L NE group, and 100 μmol/L NE group were cultured in low-sugar Dulbecco′s modified eagle medium containing 1% volume fraction of fetal bovine serum (hereinafter referred to as low-serum medium) added with NE in final molarity of 1 μmol/L, 10 μmol/L, and 100 μmol/L, respectively. Cells in PBS group were cultured in low-serum medium added with the same volume of PBS. Before stimulation (0 d) and on stimulation day 1, 3, 5, cell counting kit 8 method was used to detect cell proliferation activity (expressed as the absorbance value). (2) In cell scratch test 1, cells were divided into PBS group and simple NE group. After the scratch test, cells in simple NE group were cultured with low-serum medium+ NE in final molarity of 10 μmol/L, and cells in PBS group were cultured with low-serum medium+ the same volume of PBS. In cell scratch test 2, cells were divided into PBS group, propranolol+ NE group, and phentolamine+ NE group. After the scratch test, cells in propranolol+ NE group were pretreated with low-serum medium+ propranolol in final molarity of 1 μmol/L for 30 minutes each day, cells in phentolamine+ NE group were pretreated with low-serum medium+ phentolamine in final molarity of 10 μmol/L for 30 minutes each day, and then they were cultured with low-serum medium+ NE in final molarity of 10 μmol/L. Cells in PBS group were cultured with low-serum medium+ the same volume of PBS. In cell scratch test 3, cells were divided into simple NE group, simple (2E, 6E)-2, 6-bis (4-pyridylmethylene) cyclohexanone (SC-66) group, and SC-66+ NE group. After the scratch test, cells in simple NE group was cultured with low-serum medium+ NE in final molarity of 10 μmol/L, cells in simple SC-66 group were cultured with low-serum medium after being pretreated with SC-66 in final molarity of 30 mmol/L for 30 minutes every day, cells in SC-66+ NE group were cultured with low-serum medium+ NE in final molarity of 10 μmol/L after being pretreated with SC-66 in final molarity of 30 mmol/L for 30 minutes every day. In the above 3 cell scratch tests, the sample numbers in each group were all 6, and the scratch healing rates at post scratch hour (PSH) 24, 48, and 72 were all calculated. (3) Cells were divided into PBS group, simple NE group, propranolol+ NE group, and phentolamine+ NE group, with 3 wells in each group. The lower chamber treatment methods of PBS group and simple NE group were the same as those of the same groups in cell scratch test 1. The lower chamber treatment of propranolol+ NE group and phentolamine+ NE group were the same as those of the same groups in cell scratch test 2. After the Transwell experiment was performed and the cells were routinely cultured for 24 hours, the migrated cells were counted. (4) Cells were divided into PBS group, simple NE group, propranolol+ NE group, and phentolamine+ NE group, with 2 dishes in each group. The cell treatment of PBS group and simple NE group were the same as those of the same groups in cell scratch test 1. The cell treatment of propranolol+ NE group and phentolamine+ NE group were the same as those of the same groups in cell scratch test 2. After 24 hours of routine culture, the phosphorylation level of protein kinase B (Akt) of cells was detected by Western blotting. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, independent sample t test, least significant difference t test, and Bonferroni correction. Results (1) After 1 day of stimulation, the absorbance value of cells in 100 μmol/L NE group was significantly lower than that in PBS group (t=2.986, P<0.05). After 5 days of stimulation, the absorbance value of cells in 10 μmol/L NE group was significantly higher than that in PBS group (t=3.547, P<0.01). (2) In cell scratch test 1, at PSH 24, 48, and 72, the scratch healing rates of cells in simple NE group were (34.4±3.4)%, (52.5±4.7)%, and (70.0±3.8)%, which were significantly lower than (44.1±4.2)%, (80.0±3.6)%, and (95.9±2.2)% in PBS group (t=19.320, 128.319, 221.575, P<0.01). In cell scratch test 2, at PSH 24, 48, and 72, the scratch healing rates of cells in propranolol+ NE group were significantly lower than those in PBS group (t=4.073, 9.618, 15.272, P<0.01). In cell scratch test 3, at PSH 72, the scratch healing rates of cells in NE group was significantly lower than that in simple SC-66 group (t=8.862, P<0.01). At PSH 24, 48, and 72, the scratch healing rates of cells in SC-66+ NE group were significantly lower than those in simple SC-66 group (t=3.862, 4.290, 10.357, P<0.01). (3) The Transwell experiment showed that after 24 hours of culture, the numbers of migrated cells in simple NE group, propranolol+ NE group, and phentolamine+ NE group were significantly less than the number in PBS group (t=11.895, 10.196, 3.222, P<0.01). (4) After 24 hours of culture, the phosphorylation levels of Akt of cells in simple NE group and propranolol+ NE group were significantly higher than the level in PBS group (t=8.186, 5.996, P<0.01). Conclusions NE can inhibit the migration of BMSCs in mice, a process in which the signal pathway of Akt is involved in its regulation.

Effects of Astragaloside Ⅳ on exosome secretion and its microRNA-126 expression in human endothelial progenitor cells

Xiong Wu, Bai Xue, Xiao Hui, Lan Hongwei, Zhu Chenhong, Zhao Shiqing, Wu Yujuan, Chen Jia

2020, 36(12): 1183-1190. doi: 10.3760/cma.j.cn501120-20191222-00466

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Objective To investigate the effects of Astragaloside Ⅳ on the secretion of exosomes in human endothelial progenitor cells (EPCs) and the expression of microRNA (miRNA)-126 in exosomes. Methods The umbilical cord blood from one healthy full-term newborn from the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine in 2019 was harvested for isolating mononuclear cells by density gradient centrifugation and cultured for 7 days. Morphological observation was performed during this period. Cells of the third passage were collected for identification by CD31 immunomagnetic bead sorting and double fluorescence staining. According to the random number table, the identified EPCs were divided into Astragaloside Ⅳ group and phosphate buffer solution (PBS) group. The cells in Astragaloside Ⅳ group were cultured with Astragaloside Ⅳ in final mass concentration of 100 mg/L for 24 hours, and the cells in PBS group were cultured with the same volume of PBS for 24 hours. After culture, the exosomes from the cell culture supernatant of the two groups were collected, and the expressions of characteristic markers of exosomes CD9, CD63, and CD81 were detected by Western blotting, the morphology of EPC exosomes (EPC-Exos) was observed under transmission electron microscope, and the particle size of EPC-Exos was detected by nanoparticle tracking analysis technique. The concentration of EPC-Exos was determined by dioctyl butyric acid method (the sample number was 3), and the expressions of miRNA-126-3p and miRNA-126-5p related to angiogenesis in EPC-Exos were determined by reverse transcription polymerase chain reaction (the sample number was 3). Data were statistically analyzed with independent sample t test. Results (1) On the 4th day of culture, the cells began to adhere to the wall, and the multi-forms such as circle, fusiform, and strip appeared at the same time. On the 7th day of culture, the edge of the cells was clear and arranged like a paving stone, the central cells were round, and the surrounding cells were fusiform. (2) CD31 immunomagnetic beads sorting method identification showed that the membrane was stained with green fluorescence and the nucleus was stained with blue fluorescence. Double fluorescence staining method showed that the cells were orange-yellow. The cells were identified as EPCs. (3) After 24 hours of culture, the expressions of CD9, CD63, and CD81 in EPC-Exos were all positive, confirming that EPC-Exos were extracted successfully in this experiment. (4) After 24 hours of culture, the EPC-Exos of the two groups showed round membrane vesicles, and there was no significant difference in morphology. (5) After 24 hours of culture, the particle size of 98.7% EPC-Exos in Astragaloside Ⅳ group was 84.7 to 143.1 nm, and that of 98.0% EPC-Exos in PBS group was 88.7 to 123.5 nm. (6) After 24 hours of culture, the mass concentration of EPC-Exos in Astragaloside Ⅳ group was (310±5) μg/mL, which was significantly higher than (257±5) μg/mL in PBS group, t=13.369, P<0.01. (7) After 24 hours of culture, there were more miRNA-126-3p (t=16.062, P<0.01) and miRNA-126-5p (t=3.252, P<0.05) in EPC-Exos of Astragaloside Ⅳ group than in PBS group. Conclusions Astragaloside Ⅳ can improve the function of human EPC secretory exosomes, and the secreted exosomes are loaded with miRNA-126.

Meta-analysis of the effects of triamcinolone acetonide alone and in combination with 5-fluorouracil for treating keloids

Liu Xinjian, Cui Zhengjun, Zhang Shutang, Su Weiguo, Meng Qingnan, Guo Pengfei, Wei Aizhou, Zhou Jian, Wang Changyin, Zou Shibo, Sun Jialin, Wang Xu

2020, 36(12): 1191-1198. doi: 10.3760/cma.j.cn501120-20190930-00390

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Objective To compare the efficacy and safety of triamcinolone acetonide (TA) alone and in combination with 5-fluorouracil (5-FU) for treating keloids using meta-analysis. Methods Databases including PubMed, Embase, and Cochrane Library were retrieved with the search terms of " triamcinolone acetonide, 5-fluorouracil, glucocorticoid, fluorouracil, keloid, scar, TAC, 5-FU, hypertrophic scar " and databases including Chinese Journal Full-Text Database, Chinese Biomedical Database, and Wanfang Data were retrieved with the search terms of "曲安奈德,瘢痕疙瘩, 5-氟尿嘧啶,糖皮质激素,增生性瘢痕" in Chinese to obtain the publicly published randomized controlled trials about the effects of TA alone and in combination with 5-fluorouracil for treating keloids from the establishment of each database to august 2019. The outcome indexes included effective proportion of treatment, incidence proportion of adverse reactions, and recurrence proportion of keloids. RevMan 5.3 and Stata 14.0 statistical software were used to conduct a meta-analysis of eligible studies. Results A total of 1 326 patients with keloids were included in 14 studies, including 668 patients in TA+ 5-fluorouracil group whose keloids were injected with TA and 5-fluorouracil and 658 patients in TA alone group whose keloids were injected with TA alone. A total of 7 articles achieved 1 to 3 points in modified Jadad score, while 7 articles achieved 4 to 7 points in modified Jadad score. Patients in TA+ 5-fluorouracil group had a higher effective proportion of treatment than that of TA alone group (relative risk=1.28, 95% confidence interval=1.16-1.41, P<0.01). Subgroup analysis showed that the quality of the included literature and ethnic factors might be the source of heterogeneity in effective proportion of treatment. Patients in TA+ 5-fluorouracil group had a lower incidence proportion of adverse reactions than that of TA alone group (relative risk=0.44, 95% confidence interval=0.25-0.75, P<0.01). Patients in TA+ 5-fluorouracil group had a lower recurrence proportion of keloids than that of TA alone group (relative risk=0.25, 95% confidence interval=0.14-0.44, P<0.01). There was no publication bias in incidence proportion of adverse reactions (P>0.05), while the effective proportion of treatment and recurrence proportion of keloids had publication bias (P<0.05). Conclusions TA combined with 5-fluorouracil is more effective than TA alone for treating keloids, with less incidence of adverse reactions and recurrence.

Methods of repairing large soft tissue defect with latissimus dorsi myocutaneous flap and management of secondary wound in donor site

Ma Chao, Tao Ran, Shu Jun, Lei Yonghong, Han Yan

2020, 36(12): 1199-1203. doi: 10.3760/cma.j.cn501120-20191121-00439

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Objective To explore the methods of repairing large soft tissue defect with latissimus dorsi myocutaneous flap and the management of secondary wound in donor site. Methods From June 2015 to June 2019, 30 patients with soft tissue defect caused by various reasons or hyperplastic scar were hospitalized in the First Medical Center of Chinese PLA General Hospital, including 10 males and 20 females, aged 25-64 years, with 18 cases of head soft tissue defects caused by the growth and rupture of tumor, 7 cases of hypertrophic scar in trunk and limbs, and 5 cases of facial and neck soft tissue defects caused by trauma. The area of primary wound after debridement or enlarged lesion resection was 14 cm×10 cm-18 cm×16 cm. Preoperative evaluation of 20 patients showed that the wound was relatively large, and the donor site could not be directly closed by suturing after resection of conventional single-lobe latissimus dorsi myocutaneous flap, so the bilobed latissimus dorsi myocutaneous flap with area of 14 cm×5 cm-18 cm×8 cm was cut to repair the wound, and the donor site was directly closed by suturing. Preoperative evaluation of 10 patients showed that the donor site could be directly closed by suturing after resection of conventional single-lobe latissimus dorsi myocutaneous flap, so that conventional single-lobe latissimus dorsi myocutaneous flap with area of 11 cm×9 cm-13 cm×10 cm was resected to repair the primary wound, resulting in big tension in donor site and secondary wound with area of 6 cm×4 cm-8 cm×6 cm that couldn′t be directly sutured, which was repaired with donor site local flap with area of 7 cm×4 cm-9 cm×6 cm, and the second donor site was directly closed by suturing. Intraoperative end-to-end anastomosis was performed between the thoracodorsal arteries and veins of the latissimus dorsi myocutaneous flap and the arteries and veins of the primary recipient wound. The survival of latissimus dorsi myocutaneous flaps and local flaps were observed after surgery, and the appearance and function of the donor and recipient areas were observed during follow-up. Results All the latissimus dorsi myocutaneous flaps and local flaps survived in the patients after surgery. Follow-up of 6-12 months showed that the latissimus dorsi myocutaneous flap was similar in color to the surrounding normal skin, with soft texture and good elasticity. The donor site of 20 patients repaired with bilobed latissimus dorsi myocutaneous flaps were only left with linear scars, among which 2 patients had hypertrophic scars and none had functional impairment. The donor site of 10 patients repaired with single-lobe latissimus dorsi myocutaneous flaps and donor site local flaps had good appearance, left with linear scar, irregular shape, but no local traction or dysfunction. Conclusions When repairing a large soft tissue defect, the bilobed latissimus dorsi myocutaneous flap or the single-lobe latissimus dorsi myocutaneous flap combined with the local flap transfer in the donor site can be used after preoperative evaluation so that the donor site wound can be closed at one time while repairing the primary wound. The donor site has less scar, and both the recipient and donor sites have good appearance and function after surgery.

Application effect of sustainable skin-stretching device in scalp and soft tissue defect

Yuan Bo, Liang Haidong, Tong Zhihong, Song Wenji, Ju Shanglian

2020, 36(12): 1204-1207. doi: 10.3760/cma.j.cn501120-20200215-00061

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Objective To explore the application effect of sustainable skin-stretching device in scalp and soft tissue defect. Methods From June 2017 to January 2020, 5 patients (3 males and 2 females, aged 31-57 (38.0±2.1) years) with large area of scalp and soft tissue defect and skull exposure were admitted to Department of Reparative and Reconstructive Surgery of the Second Hospital of Dalian Medical University. The wound area ranged from 16.0 cm×8.0 cm to 18.0 cm×12.0 cm. The sustainable skin-stretching device was installed after debridement operation for scalp wound. The wound was stretched from the 3rd day after installation of the device, at a basic speed of 1 mm/d and finished for 3 times on average. During stretching, close attention was paid to the changes in blood flow of the wound margin and the subjective feeling of the patients. When the result was negative in the squeezing and pinching test for wound margin after stretching, the further stretching was stopped, the final stretching state was maintained for 3 days, and the wounds were sutured directly. The wound healing during stretching of sustainable skin-stretching device and the occurrence of complications were observed. The rest wound areas after stretching for 5, 10, 15, and 20 days were measured. The wound healing and hair growth were observed during follow-up. Results All the wounds of 5 patients was sutured directly after stretching for 19-23 d. There was no tension blister on the margin of wounds during stretching, and the margin of wounds healed well after being sutured without skin necrosis. After stretching treatment for 5-20 d, the wound areas were gradually decreased. During follow-up of 2-11 (4.5±1.5) months, the elasticity, color, feeling, and regenerated hair growth of the stretched scalp tissue were close to those of the surrounding normal skin tissue. The linear scar formed on the margin of wounds, but no scar formed on the wounds. Conclusions The application of sustainable skin-stretching device can reduce the difficulty in repairing scalp and soft tissue defect, with the regenerated hair growing well after treatment, which is worthy of clinical promotion.

Research advances on application of single-cell RNA sequencing in islet cell biology

Liu Xinzhu, Peng Yizhi, Shen Chuan′an

2020, 36(12): 1208-1212. doi: 10.3760/cma.j.cn501120-20191127-00445

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2020 Vol. 37, No. 12

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