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Clinical effects of thin anterolateral thigh perforator flaps combined with finger splitting and finger web plasty in sequential treatment of destructive wounds of total hand degloving
Yin Shanqing, Zhu Feng, Huang Yaopeng, Pan Jiadong, Xiao Dongchao, Liu Linhai, Li Xueyuan, Wang Xin
, Available online  , doi: 10.3760/cma.j.cn501225-20240723-00275
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Abstract:
  Objective  To investigate the clinical effects of thinned anterolateral thigh perforator flaps combined with finger splitting and finger web plasty in sequential treatment of destructive wounds of total hand degloving.  Methods  A retrospective observational study was conducted. From January 2012 to January 2023, a total of 15 cases who met the inclusion criteria with destructive wounds of total hand degloving were treated in Ningbo No.6 Hospital, including 10 males and 5 females, aged 17-75 years. The wounds were all combined with exposed bones and tendon. Debridement and vacuum sealing drainage were performed in all cases before flap transplantation in stage Ⅰ. After complete debridement, the wound area was 11.0 cm×3.0 cm-23.0 cm×13.5 cm. One or both anterolateral thigh perforator flaps with size of 12.5 cm×5.0 cm-25.0 cm×15.5 cm were designed and cut according to the defect area, and the flaps were thinned to repair the skin and soft tissue defects of the hand. The donor site was sutured directly or repaired with medium-thickness skin graft from the opposite thigh. According to the need, the flap was reconstructed by finger split and finger web plasty once or more times every 3 months after the flap transplantation. The survival and complications of flap and wound healing at the donor site were observed during the follow-up. The appearance of flap, two-point discrimination distance, and hand function were followed up after stage Ⅰoperation. At the final follow-up, the function of the affected hand was evaluated by the trial standards for evaluation of partial function of upper extremity by the Hand Surgery Society of Chinese Medical Association.  Results  After the operation of stage Ⅰ, all the 15 cases of patients with flap survived completely, including 1 case with arterial crisis of flap, which survived completely after exploring and re-anastomosis of blood vessels; all the wounds at the donor site healed. Follow-up for 6 to 18 months after stage Ⅰ, the flap was slightly swollen, with a little pigmentation, and the two-point discrimination distance in the finger flap was 8-11 mm. The fingers can complete the basic life functions such as flexion, extension, pinch, and grip. At the last follow-up, 3 cases were excellent, 9 cases were good, and 3 cases were acceptable.  Conclusions  For destructive wounds of total hand degloving, free transplantation of one or both thinned anterolateral thigh perforator flaps is used for repair in stage Ⅰ, and finger split and finger web plasty is used to repair the flaps in the later stage, which can basically restore the pinch and grip function of the affected hand, and is worthy of clinical promotion.
Role and mechanism of ferroptosis in acute lung injury associated with combined burn-blast trauma in rats
Zhang Hao, Guan Hao, Wang Yuhang, Zhang Wanfu, Tian Linqiang, Ren Wenjie
, Available online  , doi: 10.3760/cma.j.cn501225-20240528-00199
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  Objective  To investigates the role and mechanism of ferroptosis in acute lung injury associated with combined burn-blast trauma in rats.  Methods  This was an experimental study. Twenty-four 8-week-old male Sprague-Dawley rats were randomly divided into control group and experimental group, each containing 12 animals. The rats in experimental group were anesthetized and treated with explosion to make the model of acute lung injury associated with combined burn-blast trauma, whereas the rats in control group underwent sham injury. Twenty-four hours post-injury, the pathological morphology of lung tissue was observed by hematoxylin-eosin staining and immunohistochemical staining methods. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 in the supernatant of bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay. The arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2) of abdominal aortic blood were measured by automatic animal blood gas analyzer. The lung tissue was weighed and the dry-wet ratio was calculated. The total protein concentration in BALF was measured by bicinchoninic acid protein concentration determination kit. The levels of oxidative stress factors, such as reactive oxygen species, malondialdehyde, superoxide dismutase (SOD), glutathione, and ferrous ion in lung tissue homogenates of rats were detected by related kits. The expression of ferroptosis related molecule glutathione peroxidase 4 (GPX4), lipid peroxides related molecule 4-hydroxynonenal (4-HNE) and DNA oxidative damage related molecule 8-hydroxydeoxyguanosine (8-OHdG) in lung tissue were detected by immunofluorescence and immunohistochemistry methods. Mitochondrial morphology was observed under transmission electron microscopy.  Results  Twenty-four hours post-injury, the lung tissue structure of rats in control group was clear and complete, and the alveolar wall was normal; in experimental group, the lung tissue edema of rats was obvious, the alveolar wall became thicker, and the structure was not clear. Twenty-four hours post-injury, compared with those in control group, the levels of TNF-α, IL-1β, and IL-6 in BALF supernatant of experimental group were significantly increased (with t values of 3.96, 9.84, and 10.60, respectively, P<0.05); the dry-wet ratio of lung tissue, lung injury score, and total protein concentration in BALF of rats in experimental group were significantly increased (with t values of 6.91, 6.64, and 10.04, respectively, P<0.05), PaO2 of abdominal aortic blood decreased significantly (t=8.85, P<0.05) while PaCO2 did not change significantly (t=0.20, P>0.05); the levels of SOD and glutathione in the lung homogenate of rats in experimental group were significantly decreased (with t values of 4.36 and 8.56, respectively, P<0.05), while the average levels of reactive oxygen species, malondialdehyde, and ferrous ion were significantly increased (with t values of 11.55, 9.78, and 14.77, respectively, P<0.05). Twenty-four hours post-injury, immunofluorescence staining and immunohistochemical staining showed that the expression of GPX4 in lung tissue of rats in experimental group were 0.245±0.024 and 0.786±0.240, respectively, which were significantly lower than 1.000±0.305 and 1.000±0.200 in control group (with t values of 6.05 and 2.60, respectively, P<0.05). The expression values of 4-HNE in lung tissue of rats in experimental group were 5.93±1.05 and 2.21±0.23 by immunofluorescence and immunohistochemical staining, respectively, which were significantly higher than 1.00±0.29 and 1.00±0.23 in control group (with t values of 11.13 and 9.16, respectively, P<0.05). The expression values of 8-OHdG in lung tissues of rats in experimental group were 2.08±0.40 and 1.61±0.29 by immunofluorescence and immunohistochemical staining, respectively, which were significantly higher than 1.00±0.40 and 1.00±0.26 in control group (with t values of 4.72 and 3.87, respectively, P<0.05). Twenty-four hours post-injury, compared with that in control group, the density of the double membrane of mitochondria in the lung tissue cells of rats in experimental group increased, the outer membrane ruptured, and the crista decreased  Conclusions  In rats with acute lung injury associated with combined burn-blast trauma in rats, there is oxidative DNA damage in lung tissue cells, the imbalance of antioxidant system in lung tissue, and the expression of GPX4, the key molecule against ferroptosis, is decreased, suggesting that ferroptosis is involved in the pathophysiological process.
Respiratory care for burn-related lung injury--shoulder heavy responsibilities, and a long way to go
Zhu Feng, Guo Guanghua
, Available online  , doi: 10.3760/cma.j.cn501225-20240802-00291
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Burns is often accompanied by respiratory tract and lung injuries, which seriously affects the prognosis and treatment effect of such patients. Respiratory therapy refers to the prevention, evaluation, diagnosis, treatment, management, and care of patients with cardiopulmonary insufficiency or abnormalities under the guidance and cooperation of professional doctors, which is an important part of acute respiratory distress syndrome treatment, and also gradually applied in burn-related lung injuries. Respiratory therapy covers prevention and treatment, and interacts with burn wound repair and overall rehabilitation, which requires more attention, research, and practice from practitioners. This paper, under the background of the rapid development of respiratory therapy and the relatively limited burn related lung injury research and practice, tries to elaborate and analyze the significance of respiratory therapy for burn related lung injury, specific measures, and shortcomings, aiming to encourage more practitioners to study and participation, through accumulation and summary, to achieve the purpose of scientific and standardized implementation of respiratory care for burn-related lung injury.
Comparative study on patients with burn-blast combined injuries caused by liquid plastic and liquid metal foreign objects in the body
Ke Jiaxiang, Yu Fengge, Li Ning, Bian Xi, Deng Ruomeizi, Li Bei
, Available online  , doi: 10.3760/cma.j.cn501225-20240327-00107
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  Objective  To compare the clinical characteristics, diagnostic plans, and treatment strategies of patients with burn-blast combined injuries caused by liquid plastic and liquid metal foreign objects in the body.  Methods  This study was a retrospective cohort study. From January 2009 to July 2019, 41 patients with burn-blast combined injuries caused by hot solution explosion who met the inclusion criteria were admitted to Department of Burns and Plastic Surgery of Qingdao Hospital ofRehabilitation University. The following indexes of all patients were collected, including gender, age, total burn area, admission time after injury, site of burn-blast combined injuries, and type of foreign objects in the body. According to the type of foreign objects in the body, the patients were divided into liquid plastic group (30 cases) and liquid metal group (11 cases). The following indexes of the two groups of patients were collected, including the clinical characteristics (swelling in the injury site pain, fever, abscess formation, depth of injury, activity of foreign objects, and difficulty in removing foreign objects), imaging examinations (ultrasound, computed radiography, computed tomography, and magnetic resonance imaging examinations), treatment (repair period and repair method), and incidence of complications during follow-up after discharge.  Results  There were 33 males and 8 females among the patients, aged 18-65 years. The total burn area was 1% to 78% total body surface area, the admission time after injury was 2 h to 7 d, the burn-blast combined injuries mainly occurred in the limbs and trunk, and the foreign objects in the body were liquid plastics (polyethylene and acrylonitrile butadiene styrene) and liquid metals (liquid iron and aluminum). The proportions with swelling in the injury site, injury with depth to the bone, poor mobility of foreign objects, and difficulty in removing foreign objects in patients in liquid plastic group was significantly higher than those in liquid metal group (P<0.05). The proportions of confirming foreign objects in the body through computed radiography and computed tomography examinations in patients in liquid metal group were 7/7 and 8/8, respectively, which were significantly higher than 1/5 and 3/20 in liquid plastic group (P<0.05); the proportions of confirming foreign objects through ultrasound and magnetic resonance imaging examinations in patients in liquid metal group were 11/11 and 2/2, respectively, which were similar to 24/26 (P>0.05) and same to 4/4 in liquid plastic group. The proportions of patients in liquid plastic group who underwent stage Ⅰ wound repair and direct suture were significantly lower than those in liquid metal group (P<0.05), while the proportions of patients who underwent delayed wound repair and skin grafting were significantly higher than those in liquid metal group (P<0.05). There were no statistically significant differences in the incidence of complications between the two groups of patients (P>0.05).  Conclusions  Patients with burn-blast combined injuries may experience varying degrees of damage due to different types and locations of foreign objects in the body. Computed radiography and computed tomography examinations can be used to diagnose liquid metal foreign objects, while ultrasound and computed radiography examinations can be used to diagnose liquid plastic foreign objects in the body. Multidisciplinary collaboration and comprehensive treatment are important means of treating such kind of patients.
A multicenter study on the effect of the early infusion rate on prognosis and the influential factors of the infusion rate in patients with severe burns and inhalation injury
Huang Shengyu, Ma Qimin, Wang Yusong, Tang Wenbin, Chu Zhigang, Xin Haiming, Chang Liu, Li Xiaoliang, Guo Guanghua, Zhu Feng
, Available online  , doi: 10.3760/cma.j.cn501225-20240409-00130
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  Objective  To investigate the effect of the early infusion rate on prognosis and the influential factors of the infusion rate in patients with severe burns and inhalation injury.  Methods  This study was a multicenter retrospective case series research. From January 2015 to December 2020, 220 patients with severe burns and inhalation injury meeting the inclusion criteria were admitted to 7 burn treatment centers in China, including 13 cases in the Fourth People's Hospital of Dalian, 26 cases in the First Affiliated Hospital of Naval Medical University, 73 cases in the Guangzhou Red Cross Hospital of Jinan University, 21 cases in the 924th Hospital of PLA, 30 cases in the First Affiliated Hospital of Nanchang University, 30 cases in the Tongren Hospital of Wuhan University ＆ Wuhan Third Hospital, and 27 cases in Zhengzhou First People's Hospital. There were 163 males and 57 females, and their ages ranged from 18 to 91 years. The patients were divided into survival group and death group according to the survival within 28 d post injury. The following data of patients in the 2 groups were collected, including basic information (gender, age, body weight, body temperature, and etc.), the injury characteristics (total burn surface area, post-injury admission time, and etc.), the underlying diseases, the post-injury fluid resuscitation condition (infusion rate and ratio of infused electrolyte solution to colloid solution in the first 24 h post injury, and etc.), the results of laboratory tests on admission (blood urea nitrogen, blood creatinine, albumin, pH value, base excess, blood lactate, oxygenation index on admission, and etc.), and treatment condition (inhaled oxygen volume fraction, hospitalization day, renal replacement therapy, and etc.). After adjusting covariates using univariate Cox regression analysis, the multivariate COX regression was performed to analyze the effect of infusion rate in the first 24 h post injury on death. The receiver operator characteristic curve for the infusion rate in the first 24 h post injury to predict the risk of death was plotted, and the maximum Youden index was calculated. Patients were divided into 2 groups according to the cutoff value for predicting risk of death by the infusion rate in the first 24 h post injury determined by the maximum Youden index, and the risk of death was compared between the 2 groups. The correlation between the previously mentioned clinical data and the infusion rate in the first 24 h post injury was analyzed; after the univariate linear regression analysis was used to screen the independent variables, the multivariate linear regression analysis was performed to screen the independent influential factors on the infusion rate in the first 24 h post injury.  Results  Compared with those in survival group, patients in death group had significantly higher age and total burn surface area (with Z values of 12.08 and 23.71, respectively, P<0.05), the infusion rate, inhaled oxygen volume fraction, blood urea nitrogen, blood creatinine, and blood lactic acid in the first 24 h post injury (with Z values of 7.99, 4.01, 11.76, 23.24, and 5.97, respectively, P<0.05), and the proportion of patients treated with renal replacement therapy (P<0.05) were significantly higher, the albumin, pH value, and base excess were significantly lower (t=2.72, 8.18 and with Z values of 9.70, respectively, P<0.05), and the hospitalization day was significantly shorter (Z=85.47, P<0.05). After adjusting covariates, the infusion rate in the first 24 h post injury was the independent influential factor on death (with standardized hazard ratio of 1.69, 95% confidence interval of 1.21-2.37, P<0.05). Patients in infusion rate ≥2.03 mL·kg-1·% total body surface area (TBSA)-1 group had a significantly higher risk of death than those in infusion rate <2.03 mL·kg-1·% TBSA-1 group (with hazard ratio of 3.47, 95% confidence interval of 1.48-8.13, P<0.05). There was a significant correlation between total burn surface area, body weight, inhaled oxygen volume fraction, body temperature, post-injury admission time, the ratio of infused electrolyte solution to colloid solution in the first 24 h post injury, and oxygenation index <300 on admission and the infusion rate in the first 24 h post injury (with r values of -0.19, -0.21, 0.14, -0.16, -0.25, and 0.31, respectively, Z=4.48, P<0.05). Total burn surface area, body weight, post-injury admission time, and oxygenation index <300 on admission were the independent influential factors on the infusion rate in the first 24 h post injury (with standardized β values of -0.22, -0.22, -0.19, and 0.46, respectively, 95% confidence intervals of -0.34 to 0.09, -0.34 to 0.10, -0.32 to 0.06, and 0.22 to 0.71, respectively, P<0.05).  Conclusions  The infusion rate in the first 24 h post injury in patients with severe burns and inhalation injury was the independent influential factor on death, and patients with infusion rate ≥2.03 mL·kg-1·%TBSA-1 in the first 24 h post injury have a significantly increased risk of death. The total burn surface area, body weight, post-injury admission time, and oxygenation index <300 on admission were the independent influential factors of the infusion rate in the first 24 h post injury.
The role and mechanism of human umbilical cord mesenchymal stem cell exosomes in wounds with scabbing and skin grafting in scalded rats
Wang Di, Dou Shuqian, Wu Kongjia, Zhang Gaofei, Lou Hanxiao, Zhang Chenying, Yang Guoxun, Jin Chengbo, Que Ting, Liu Wenjun
, Available online  , doi: 10.3760/cma.j.cn501225-20231201-00223
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  Objective  To investigate the role and mechanism of human umbilical cord mesenchymal stem cell exosomes (hUCMSC-ex) in wounds with scabbing and skin grafting in scalded rats.  Methods  The study was an experimental study. Twelve male Sprague-Dawley rats aged 6-8weeks were divided into combined treatment group, fixed+allogeneic skin group, autologous skin+allogeneic skin group, and allogeneic skin group by random number table method (the same rats and grouping methods were used hereafter), with 3 rats in each group. The four groups of rats were inflicted with scalded wounds on the back and with scabbing, and then the wounds of rats in combined treatment group were fixed with a metal ring and transplanted with autologous skin grafts and allogeneic skin grafts, and the other three groups were fixed and/or transplanted with skin grafts corresponding to the group name. On 14, 21, and 28 d after surgery, the wound healing area in four groups of rats was measured. Another 15 rats were divided into normal group with no treatment, high exosome group, low exosome group, supernatant group, and phosphate buffered solution (PBS) group, with 3 rats in each group. The last 4 groups of rats were treated as that in the above-mentioned combined treatment group, and then were injected around the wounds with 200 μL of PBS containing 100 μg of hUCMSC-ex, 200 μL of PBS containing 50 μg of hUCMSC-ex, supernatant with no hUCMSC-ex, and PBS at 0 (immediately), 7, 14, and 21 d after surgery, respectively. On 14, 21, and 28 d after surgery, the wound healing area in four groups of rats was measured. The wound neo-epithelial tissue of rats in high exosome group and PBS group on 28 d after surgery and the normal skin tissue of rats in normal group at the same time point were taken, and the differentially expressed proteins were screened by label-free quantitative proteomics technology; the protein expressions of IgG1 heavy chain constant region (IGHG1) and cystatin A (CSTA) which were the differentially expressed proteins with the most obvious differences between high exosome group and PBS group were detected by Western blotting. The number of samples in all experiments was 3.  Results  On 14, 21, and 28 d after surgery, the wound healing area in combined treatment group, autologous skin+allogeneic skin group and allogeneic skin group of rats was significantly larger than that in fixed+allogeneic skin group (P<0.05), the wound healing area in autologous skin+allogeneic skin group of rats on 21 d after surgery and that in allogeneic skin group of rats on 14 and 21 d after surgery was significantly larger than that in combined treatment group (P<0.05), and the wound healing area in allogeneic skin group of rats on 14 d after surgery was significantly larger than that in autologous skin+allogeneic skin group (P<0.05). The wound healing area of rats in high exosome group and low exosome group on 14, 21, and 28 d after surgery and in supernatant group on 14 and 28 d after surgery were significantly larger than those in PBS group (P<0.05); the wound healing area in high exosome group of rats on 14 and 28 d after surgery was significantly larger than that in supernatant group (P<0.05), and that on 14 d after surgery was significantly larger than low exosome group (P<0.05); the wound healing area in low exosome group of rats on 14 d after surgery was significantly larger than that in supernatant group (P<0.05). Compared with that in PBS group, there were 332 differentially expressed proteins in the wound neo-epithelial tissue in high exosome group on 28 d after surgery (P<0.05), among which IgG1 heavy chain constant region IGHG1 and CSTA proteins were significantly up-regulated (with foldchange of 12.60 and 2.27, respectively, P<0.05). Compared with those of normal skin tissue in normal group, the differentially expressed proteins in the wound neo-epithelial tissue in high exosome group and PBS group of rats on 28 d after surgery were 1 400 and 1 057, respectively. The protein expressions of IGHG1 and CSTA in the wound neo-epithelial tissue in high exosome group of rats on 28 d after surgery were significantly larger than those in PBS group (P<0.05) and those of normal skin tissue of rats in normal group (P<0.05).  Conclusions  hUCMSC-ex may accelerate the repair process of wounds with scabbing and skin grafting and improve the wound healing quality in scalded rats by regulating the protein expressions of IGHG1 and CSTA.
Re-cognization of inhalation injuries
Guo Guanghua, Huang Shengyu
, Available online  , doi: 10.3760/cma.j.cn501225-20240616-00234
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As a critical risk factor for mortality in burn patients, inhalation injury has garnered the attention of various healthcare professionals. In recent years, while the incidence of burns with inhalation injury has been decreasing, the importance of treating inhalation injury should not be overlooked, as there is still scope for its diagnosis and treatment. With the deepening understanding of inhalation injury mechanisms and the emergence of new concepts on lung injury, it may offer novel insights and strategies for clinical diagnosis and treatment. This paper addresses some of these insights and concepts from the perspectives of molecular mechanisms, diagnostic evaluations, and treatment for the reference of our colleagues.
Strategies for the diagnosis and treatment of inhalation injuries in children
Yu Jia'ao, Zhang Xiuhang
, Available online  , doi: 10.3760/cma.j.cn501225-20240816-00310
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In the expansive field of burn medicine, inhalation injury represents a complex and challenging topic that has consistently captured the attention of the burn community. The comprehension of inhalation injury has become increasingly profound and comprehensive. The complexity of their condition and the uniqueness of their treatment options become especially significant when our attention is focused on the special group of children. The objective of this paper is to examine the intricate causes of inhalation injuries in children, with a particular emphasis on the distinctive characteristics of inhalation injuries in children and the subtle alterations in their pathophysiological mechanisms. In light of the aforementioned considerations, we will undertake an investigation into the precise diagnostic methods and efficacious therapeutic options for this type of injury. Our objective is to provide a foundation for the advancement and practical implementation of inhalation injuries in children, thereby facilitating the updating of knowledge and technological innovation in this domain.
Two-sample Mendelian randomization analysis of the causal relationship between inhalation injury and circulating inflammatory proteins in humans
Dai Zhanzhan, Zhu Qin, Tong Xirui, Ma Bing, Xia Zhaofan, Fang He
, Available online  , doi: 10.3760/cma.j.cn501225-20240429-00155
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  Objective  To explore the causal relationship between inhalation injury and circulating inflammatory proteins in humans.  Methods  This research was based on two-sample Mendelian randomization (MR) analysis. With inhalation injury as the exposure factor and circulating inflammatory proteins as the result, data on inhalation injury (216 993 samples) and 91 circulating inflammatory proteins (14 824 samples) were obtained from the genome-wide association study database, and analysis was conducted by two-sample MR analysis methods. Independent single nucleotide polymorphisms (SNPs) significantly associated with inhalation injury were identified as the instrumental variables using the method of linkage disequilibrium analysis. The inverse variance weighted (IVW) method was mainly used for analysis of the causal relationship between inhalation injury and 91 circulating inflammatory proteins, and further verification was carried out using the weighted median method, weighted pattern method, MR-Egger method, and simple pattern method. Based on the IVW method analysis results, SNPs of inhalation injury conformed to the hypothesis were subjected to Cochran's Q test for heterogeneity assessment, the MR-Egger regression test and the MR-PRESSO outlier test for assessment of horizontal pleiotropy, and the leave-one-out analysis for reliability assessment.  Results  Six SNPs with a significant threshold (P<5×10-5) were identified as representative instrumental variables of inhalation injury, with F values greater than 10, indicating strong correlated instrumental variables. Based on the 6 inhalation injury SNPs, the IVW method analysis revealed a significant causal relationship between inhalation injury and interleukin-20 (IL-20), IL-20 receptor subunit alpha (IL-20RA), IL-5, and tumor necrosis factor receptor superfamily member 9 (TNFRSF9), with odds ratios of 1.01, 1.01, 1.02, and 1.01, respectively, and 95% confidence intervals of 1.00-1.02, 1.00-1.03, 1.01-1.03, and 1.00-1.03, respectively, P<0.05. Results obtained via verification through the weighted median method and MR-Egger method confirmed that the causal relationships between inhalation injury and IL-5 (with odds ratios of 1.02 and 1.03 respectively, confidence intervals of 1.00-1.04 and 1.01-1.04, respectively, P<0.05) as well as TNFRSF9 (with odds ratios of 1.02 and 1.03, respectively, confidence intervals of 1.00-1.04 and 1.01-1.04, respectively, P<0.05) were statistically significant. Conversely, results obtained via verification through the weighted pattern method and simple pattern method indicated that the causal relationships between inhalation injury and IL-20, IL-20RA, IL-5, and TNFRSF9 were not statistically significant (all P values >0.05), thus reinforcing reliance on IVW method outcomes was warranted in this context. Based on the IVW method analysis results, the Cochran's Q test demonstrated no existence of significant heterogeneity among the 6 inhalation injury SNPs that had significant causal relationships with IL-20, IL-20RA, IL-5, and TNFRSF9 (with Q values of 2.67, 5.00, 5.17, and 5.29, respectively, P>0.05); assessments using the MR-Egger regression test along with MR-PRESSO outlier test confirmed that there was no significant horizontal pleiotropy among the 6 inhalation injury SNPs that had significant causal relationships with IL-20, IL-20RA, IL-5, and TNFRSF9 (with intercepts of 0.01, <0.01, -0.02, and -0.03, respectively, RSSobs values of 3.33, 9.00, 7.88, and 7.26, respectively, P>0.05); the leave-one-out analysis showed that the significant causal relationship between inhalation injury and IL-20, IL-20RA, IL-5, and TNFRSF9 was stable and reliable after removing the 6 inhalation injury SNPs one by one.  Conclusions  Two-sample MR analysis elucidates significant causal relationships between inhalation injury and four circulating inflammatory proteins, namely IL-20, IL-20RA, IL-5, and TNFRSF9, suggesting an upward trend in the generation of the above four circulating inflammatory proteins following instances of inhalation injury.
Changes in biological characteristics of adipose-derived stem cells from obese patients post successful weight loss
Wei Zhiru, Dong Yan, Qiao Gaihong, Liu Linbo, Li Guangshuai
, Available online  , doi: 10.3760/cma.j.cn501225-20231205-00225
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Abstract:
  Objective  To explore the changes in biological characteristics of adipose-derived stem cells (ASCs) from obese patients post successful weight loss, so as to provide a reference for the clinical application of these ASCs in the repair of refractory wounds.  Methods  This study was an experimental study. Twelve obese patients (8 females and 4 males, aged (50±9) years) who successfully lost weight, underwent abdominal skin tightening surgery, and were hospitalized at the First Affiliated Hospital of Zhengzhou University from April 2021 to April 2023 were included in weight loss group. During the same period, 12 healthy volunteers (10 females and 2 males, aged (50±9) years) who underwent abdominal liposuction followed by facial fat grafting in the same institution were recruited into healthy group. Adipose tissue was collected from patients in weight loss group and volunteers in healthy group, and ASCs were extracted. Experiments were conducted using ASCs at passages 4 and 5 in their logarithmic growth phase. Cell proliferation levels were assessed using the methyl thiazolyl tetrazolium assay at 0 (immediately), 24, 48, and 72 hours of culture. The cell scratch test was performed and the cell migration rates at 12 and 24 hours after scratching were calculated. The cell Transwell assay was performed and the number of migration cells at 24 hours after culture was counted. Adipogenic and osteogenic induction assays were carried out, and the adipogenic and osteogenic differentiation levels of cells were detected at 18 and 21 days of culture, respectively. Real-time fluorecence quantitative reverse transcription polymerase chain reaction was employed to measure the mRNA expressions of lipoprotein lipase (LPL), peroxisome proliferator-activated receptor gamma (PPARγ), Runt-related transcription factor 2 (Runx2), osteopontin, alkaline phosphatase (ALP), matrix metalloproteinase 9 (MMP-9), and transforming growth factor beta (TGF-β). The sample number of each experiment was 12.  Results  At 0 hour of culture, the cell proliferation levels of patients in weight loss group and volunteers in healthy group were 1.022±0.056 and 1.000±0.144, respectively, with no statistically significant difference between the groups (P>0.05). At 24, 48, and 72 hours of culture, the cell proliferation levels of patients in weight loss group were 1.366±0.030, 1.353±0.012, and 1.390±0.016, respectively, which were significantly lower than 1.755±0.077, 1.737±0.014, and 1.700±0.023 of volunteers in healthy group (with t values of 16.27, 71.35, and 38.56, respectively, P values all <0.05). At 12 and 24 hours after scratching, the cell migration rates of patients in weight loss group were lower than those of volunteers in healthy group, but the differences were not statistically significant (P>0.05). In the cell Transwell assay, after 24 hours of culture, there was no statistically significant difference in the number of migrated cells between patients in weight loss group and volunteers in healthy group (P>0.05). After 18 days of adipogenic induction, the cell adipogenic differentiation level of patients in weight loss group was significantly lower than that of volunteers in healthy group (t=27.81, P<0.05). After 21 days of osteogenic induction, the cell osteogenic differentiation level of patients in weight loss group was significantly lower than that of volunteers in healthy group (t=14.85, P<0.05). Compared with those of volunteers in healthy group, the mRNA expressions of LPL, PPARγ, TGF-β, and Runx2 of patients in weight loss group were significantly reduced (with t values of 59.48, 146.10, 46.10, and 3.13, respectively, P<0.05), while there were no statistically significant changes in the mRNA expressions of osteopontin, ALP, or MMP-9 (P>0.05).  Conclusions  Compared with healthy volunteers, ASCs from patients who successfully lose weight exhibit significantly diminished proliferative capacity, relatively weakened differentiation potential, and downregulation of some genes corresponding to adipogenic and osteogenic differentiation, which affects the therapeutic efficacy of these ASCs in treating refractory wounds caused by burns, diabetes, or radiation injuries. Therefore, in clinical application, the donor variability of ASCs needs to be considered.
Influence and mechanism of baicalin on healing of full-thickness skin defect wounds in diabetic mice
Shi Yan, Yi Liang, Zhang Weiqiang, Liu Nike, Wen Huicai, Yang Ronghua
, Available online  , doi: 10.3760/cma.j.cn501225-20231104-00179
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Abstract:
  Objective  To investigate the influence and mechanism of baicalin on the healing of full-thickness skin defect wounds in diabetic mice.  Methods  This study was an experimental research. Mononuclear cells were isolated from five male C57BL/6J mice aged 8-12 weeks and induced to differentiate into macrophages for conducting the subsequent experiments. According to the random number table (the same grouping method below), macrophages in a high-glucose environment were divided into 0 μmol/L baicalin group (no baicalin solution was added), 5 μmol/L baicalin group, 15 μmol/L baicalin group, 25 μmol/L baicalin group, 50 μmol/L baicalin group, and 75 μmol/L baicalin group treated with the corresponding final molarity of baicalin and 1 μg/mL endotoxin/lipopolysaccharide (LPS). After treatment of 48 h, the cell proliferation activity was detected using a microplate reader. Macrophages in a high-glucose environment were divided into LPS group treated with 1 μg/mL LPS and LPS+baicalin group treated with 50 μmol/L baicalin+1 μg/mL LPS. After treatment of 48 h, the percentage of double-positive cells for inducible nitric oxide synthase (iNOS) and CD80, as well as that for arginase 1 (Arg1) and CD206 among the cells, were detected using immunofluorescence method, the secretion levels of interleukin 1β (IL-1β), IL-6, IL-23, IL-10, insulin-like growth factor (IGF), and transforming growth factor β1 (TGF-β1) by the cells were measured using enzyme-linked immunosorbent assay, the expression of reactive oxygen species in the cells was detected using a fluorescent probe method, the protein expression of nuclear factor κB in the cells were detected using Western blotting, and the expression of nuclear factor 2 in the cells was observed using immunofluorescence method. The number of cell experimental samples was 3. Twenty-four 8-week-old male db/db mice (diabetic mice) were selected. After preparing full-thickness skin defect wounds on their backs, they were divided into baicalin group and normal saline group (with 12 mice in each group). On the third day after injury, 50 μmol/L baicalin and normal saline were injected into the wounds of the two groups of mice, respectively. The wound healing situation was observed and the percentage of the residual wound was calculated on the 4th, 8th, and 12th day after injury. The wound tissue was sampled on the 8th day after injury, hematoxylin-eosin staining was performed to observe the epithelial regeneration and inflammatory cell infiltration, the protein expression of CD31 was detected by Western blotting, and the expression of reactive oxygen species was detected by a microplate reader. The number of animal experimental samples was 6.  Results  After treatment of 48 h, only the proliferation activity of macrophages in 50 μmol/L baicalin group was significantly higher than that in 0 μmol/L baicalin group (P<0.05). After treatment of 48 h, the percentage of double-positive cells for iNOS and CD80 among the macrophages in LPS+baicalin group was (21.0±2.4)%, which was significantly lower than (66.6±4.5)% in LPS group (t=15.63, P<0.05); the percentage of double-positive cells for Arg1 and CD206 among the macrophages in LPS+baicalin group was (59.1±2.1)%, which was significantly higher than (18.6±1.7)% in LPS group (t=25.38, P<0.05); compared with those in LPS group, the secretion levels of IL-1β, IL-6, and IL-23 by the macrophages in LPS+baicalin group were significantly decreased (with t values of 14.26, 15.95, and 12.23, respectively, P<0.05), while the secretion levels of IL-10, IGF, and TGF-β1 were significantly increased (with t values of 8.49, 11.98, and 13.84, respectively, P<0.05); the expression of reactive oxygen species in the macrophages in LPS+baicalin group was significantly lower than that in LPS group (t=5.54, P<0.05); compared with those in LPS group, the protein expression of nuclear factor κB in the nucleus of the macrophages in LPS+baicalin group was significantly decreased (t=36.22, P<0.05), while that in the cytoplasm was significantly increased (t=14.47, P<0.05), and the expression of nuclear factor 2 in the nucleus was increased. On the 4th and 8th day after injury, the wound area of mice in baicalin group was significantly smaller than that in normal saline group, and the wounds of mice in baicalin group basically healed on the 12th day after injury. On the 4th, 8th, and 12th day after injury, the residual wound percentage of mice in baicalin group was significantly lower than that in normal saline group (with t values of 13.29, 10.08, and 11.72, respectively, P<0.05). On the 8th day after injury, compared with those in saline group, the wound tissue of mice in baicalin group showed significant re-epithelization, reduced inflammatory cell infiltration, significantly increased protein expression of CD31 (t=17.23, P<0.05), and significantly reduced expression of reactive oxygen species (t=5.78, P<0.05).  Conclusions  Baicalin reduces the inflammatory response of macrophages by lowering the level of reactive oxygen species in cells and promoting the polarization of macrophages to the M2 type, thereby facilitating the healing of full-thickness skin defect wounds in diabetic mice.

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