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Sub-cytotoxic concentrations of ionic silver promote the proliferation of human keratinocytes by inducing the production of reactive oxygen species

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Frontiers of Medicine 2018年第3期12卷 289-300页

作者： Xiaodong Duan daizhi peng Yilan Zhang Yalan Huang Xiao Liu Ruifu Li Xin Zhou Jing Liu Institute of Burn Research Southwest Hospital Third Military Medical University Chongqing 400038 China Burn and Plastic Surgery Department 209 Hospital of PLA Mudanjiang 157011 China Tissue Engineering Research Unit State Key Laboratory of Trauma Burns and Combined Injury Third Military Medical University Chongqing 400038 China

Silver-containing preparations are widely used in the management of skin wounds, but the effects of silver ions on skin wound healing remain poorly understood. This study investigated the effects of silver ions （Ag^＋） on the proliferation of human skin keratinocytes （HaCaT） and the production of intracellular reactive oxygen species （ROS）. After treating HaCaT cells with Ag^＋ and/or the active oxygen scavenger N-acetyl cysteine （NAC）, cell proliferation and intracellular ROS generation were assessed using CCK-8 reagent and DCFH-DA fluorescent probe, respectively. In addition, 5-bromo-2-deoxyUridine （BrdU） incorporation assays, cell cycle flow cytometry, and proliferating cell nuclear antigen （PCNA） immunocytochemistry were conducted to further evaluate the effects of sub-cytotoxic Ag^＋ concentrations on HaCaT cells. The proliferation of HaCaT cells was promoted in the presence of 10^-6 and 10^-5s mol/L Ag^＋at 24, 48, and 72 h. Intracellular ROS generation also significantly increased for 5 60 min after exposure to Ag^＋ The number of BrdU-positive cells and the presence of PCNA in HaCaT cells increased 48 h after the addition of 10^-6 and 10^-5 mol/L Ag^＋, with 10^-5 mol/L Ag^＋ markedly increasing the cell proliferation index. These effects of sub-cytotoxic Ag^＋ concentrations were repressed by 5 mmol/L NAC. Our results suggest that sub-cytotoxic Ag^＋ concentrations promote the proliferation of human keratinocytes and might be associated with a moderate increase in intracellular ROS levels. This study provides important experimental evidence for developing novel silver-based wound agents or dressings with few or no cytotoxicity.

关键词： ionic silver human keratinocyte cell proliferation reactive oxygen species active oxygen scavenger NAC

The progress of Chinese burn medicine from the Third Military Medical University—in memory of its pioneer,Professor Li Ao

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Burns & Trauma 2017年第2期5卷 68-78页

作者： Haisheng Li Junyi Zhou Yizhi peng Jiaping Zhang Xi peng Qizhi Luo Zhiqiang Yuan Hong Yan daizhi peng Weifeng He Fengjun Wang Guangping Liang Yuesheng Huang Jun Wu Gaoxing Luo Institute of Burn Research State Key Laboratory of Trauma Burn and Combined Injury Southwest Hospital Third Military Medical UniversityGaotanyan Street no.29 Shapingba District Chongqing 400038 China

Professor Li Ao was one of the founders of Chinese burn medicine and one of the most renowned doctors and researchers of burns in China. He established one of the Chinese earliest special departments for burns at Third Military Medical University (TMMU) in 1958. To memorialize Professor Li Ao on his 100th birthday in 2017 and introduce our extensive experience, it is our honor to briefly review the development and achievement of the Chinese burn medicine from TMMU. The epidemiology and outcomes of admitted burn patients since 1958 were reviewed. Furthermore, main achievements of basic and clinical research for the past roughly 60 years were presented. These achievements mainly included the Chinese Rule of Nine, fluid resuscitation protocol, experience in inhalation injury, wound treatment strategies, prevention and treatment of burn infections, nutrition therapy, organ support therapies, and rehabilitation. The progress shaped and enriched modern Chinese burn medicine and promoted the development of world burn medicine.

关键词： Burn injury Chinese burn medicine Li Ao Third Military Medical University Inhalation injury

Characteristics of burn deaths from 2003 to 2009 in a burn center: A retrospective study

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Burns & Trauma 2013年第2期1卷 80-86页

作者： Jian Chen Hong Yan Gaoxing Luo Qizhi Luo Xiaolu Li Jiaping Zhang Zhiqiang Yuan daizhi peng Yizhi peng Jianian Hu Jun Wu Institute of Burn Research Southwest Hospital State Key Laboratory of Trauma Burns and Combined Injury The Third Military Medical University Chongqing China

Mortality remains one of the most important end-point quality control parameters to evaluate a burn care system. We retrospectively reviewed the characteristics and multiple organ dysfunction syndrome (MODS) patterns of burn deaths in our center from January 2003 to December 2009. The mortality rate during this time period was 2.3%. Fifty-six patients died, including 49 males and 7 females. The mean survival time was 28.45 ± 24.60 days. The burn percentage was (76.70 ± 26.86) % total burn surface area (TBSA), with (27.74 ± 24.95) % deep-partial thickness burns and (46.88 ± 33.84) % full-thickness burns. Inhalation injury was diagnosed in 36 (64.29%) patients. Patients who had undergone an operation, particularly in the first week post-burn, had a significantly longer survival time. An average of 5.50 ± 1.35 malfunctioning organs per patient and a mean sequential organ failure assessment (SOFA) score of 13.91 ± 3.65 were observed. The most frequently malfunctioning organs were involved in the respiratory, hematologic, circulatory, and central nervous systems. Most of the organ damage occurred during the first week post-burn, followed by 4 weeks later, with relatively less organ damage observed in the third week. Among patients with a TBSA over 50%, non-survivors had larger burn sizes (particularly larger full-thickness burns) and a higher incidence of inhalation injury compared with survivors;nonsurvivors were also more likely to have microorganism-positive blood and sputum cultures. In conclusion, burn deaths are related to a higher burn percentage, inhalation injury, MODS, and infection. Early operation may help improve survival duration.

关键词： Burns death multiple organ dysfunction syndrome

Guideline for diagnosis, prophylaxis and treatment of invasive fungal infection post burn injury in China 2013

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Burns & Trauma 2014年第2期2卷 45-52页

作者： Gaoxing Luo Jianglin Tan Yizhi peng Jun Wu Yuesheng Huang daizhi peng Xu Wang Dahai Hu Shongtao Xi Guoan Zhang Chunmao Han Xiaoyuan Huang Ciyu Jia Jiake Chai Jingning Huan Guanghua Guo Jianhua Zhan Weiguo Xie Ying Cen Rong Yu Huade Chen Xihua Niu Yibing Wang Jinfeng Fu Baosheng Xue State Key Laboratory of Trauma Burns and Combined InjuryInstitute of Burn ResearchSouthwest Hospitalthe Third Military Medical UniversityChongqing Department of Burns and Cutaneous Surgery Xijing HospitalFourth Military Medical UniversityXi'anShanxi Department of Burns Beijing Jishuitan HospitalForth Medical College of Peking UniversityBeijing Department of Burns&Wound Care Center the Second Affiliated Hospital of Zhejiang UniversityCollege of MedicineHangzhou Department of Burns and Plastic Surgery Central South UniversityChangshaHunan Graduate School Medical College of Chinese PLABeijing Department of Burn&Plastic Surgery the First Affiliated Hospital of PLA General HospitalBeijing Department of Burn and Plastic Surgery Ruijin HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghai Department of Critical Care Medicine the First Affiliated Hospital of Nanchang UniversityNanchangJiangxi Department of Anesthesia and Critical Care Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubei Department of Otorhinolaryngology Head and Neck Surgery West China HospitalSichuan UniversityChengduSichuan Department of Burns General Hospital of Guangdong ProvinceGuangzhouGuangdong Department of Otolaryngology Henan Province HospitalZhengzhouHenan Department of Aesthetic Plastic and Burn Surgery Provincial Hospital Affiliated to Shandong UniversityJinanShandong Department of Burns Second Affiliated Hospital of Kunmin Medical UniversityYunnan Department of Burns the First Affiliated Hospital of Chinese Medical UniversityShenyangLiaoningChina

Invasive fungal infection is one of the major complication of severe burns which can induce local or systemic inflammatory response and cause serious substantial damage to the patient. The incidence of fungal infection for burn victims is increasing dramatically during recent years. This guideline, organized by Chinese Society of Burn Surgeons, aims to standardize the diagnosis, prevention and treatment of burn invasive fungal infection. It can be used as one of the tools for treatment of major burn patients.

关键词： Burn injury guideline diagnosis prophylaxis treatment invasive fungal infection

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微粒皮移植术修复深度烧伤创面的研究进展

中华损伤与修复杂志（电子版） 2016年第4期11卷 310-313页

作者：张宜澜彭代智第三军医大学西南医院全军烧伤研究所创伤、烧伤与复合伤国家重点实验室重庆400038 第三军医大学大坪医院医院野战外科研究所颌面外科重庆400038

微粒皮移植术自应用临床以来,已经成功治疗了大量严重烧伤患者,并逐渐成为大面积深度烧伤患者主要的创面修复方式之一。但是,微粒皮移植术仍然存在很多的问题,如I期愈合率相对较低,创面愈合后反复破溃,后期瘢痕增生较严重等。本文就近... 详细信息

微粒皮移植术自应用临床以来,已经成功治疗了大量严重烧伤患者,并逐渐成为大面积深度烧伤患者主要的创面修复方式之一。但是,微粒皮移植术仍然存在很多的问题,如I期愈合率相对较低,创面愈合后反复破溃,后期瘢痕增生较严重等。本文就近年来国内外学者对微粒皮移植的大量基础研究和临床观察进行了总结,重点阐述了异体微粒皮、真皮基质以及干细胞在微粒皮移植中的应用。这些方法对深度烧伤的创面愈合速度及愈后质量均有显著提高。

关键词：深度烧伤创面修复微粒皮混合移植复合移植干细胞

新生小牛皮肤的形态学和生物力学特点及脱细胞方法探索

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重庆医学 2012年第11期41卷 1043-1046,F0002页

作者：陈博彭代智左海斌郑必祥周灵第三军医大学西南医院全军烧伤研究所/创伤、烧伤与复合伤国家重点实验室重庆400038

目的通过新生小牛与成人皮肤组织在形态学和生物力学性能方面的对比研究,寻找临床应用牛真皮基质的可能证据,并初步探索对牛真皮组织进行脱细胞处理的方法。方法收集新生小牛和成人背部全层皮肤标本,采用大体观察、HE染色、Masson三色... 详细信息

目的通过新生小牛与成人皮肤组织在形态学和生物力学性能方面的对比研究,寻找临床应用牛真皮基质的可能证据,并初步探索对牛真皮组织进行脱细胞处理的方法。方法收集新生小牛和成人背部全层皮肤标本,采用大体观察、HE染色、Masson三色染色、天狼猩红染色、Gomori醛复红染色、扫描电镜和透射电镜的方法,显微照相后进行形态学观察和图像分析软件的测量;采用力学生物材料试验机检测皮肤的力学性能。采用胰蛋白酶与去污剂的不同浓度和时间组合方式对牛真皮组织进行脱细胞处理,初步观察其脱细胞效果。结果新生小牛与人的皮肤比较,真皮和表皮的厚度均明显变薄,两者的比值显著减小;真皮内Ⅰ型和Ⅲ型胶原纤维分别较人真皮增多和减少,但真皮胶原纤维束的间隙率[(41.72±1.81)∶(40.66±1.40),P=0.467]和胶原纤维束粗细[(11.28±0.18)μm∶(10.88±0.66)μm,P=0.368]差异却没有统计学意义。扫描电镜观察可见新生牛真皮胶纤维束与人真皮胶原纤维束均较纤细,排列疏松,且有一定的孔隙分布;透射电镜显示新生牛真皮胶原原纤维横纹周期长度较人真皮短,但两者的胶原原纤维直径显著差异。生物力学检测示新生牛皮肤最大应力[(21.08±0.91)MPa]和杨氏模量[(82.12±1.23)MPa]均要明显高于人皮肤的最大应力[(12.76±1.60)MPa,P=0.001]和杨氏模量[(48.63±5.50)MPa,P=0.001],而最大应变却明显低于人真皮[(0.51±0.002)mm∶(0.75±0.028)mm,P=0.001]。脱细胞方法探索显示随着胰酶浓度的增加、处理时间的延长,新生牛真皮基质内细胞核成分越来越少,但对真皮胶原结构的破坏也越来越大。结论新生牛皮肤虽然在表皮与真皮厚度、真皮胶原类型比例以及生物力学上与人皮肤有明显差异,但真皮的胶原纤维束三维结构两者之间存在较大相似性。适当浓度和时间的胰蛋白酶组合可制备出空间结构和脱细胞效果较为理想的新生牛脱细胞真皮基质,为修复人体软组织缺损提供有应用前景的生物材料。

关键词：皮肤组织学生物力学脱细胞真皮基质

Guidelines for burn rehabilitation in China

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Burns & Trauma 2015年第3期3卷 133-142页

作者： Chinese Burn Association Chinese Association of Burn Surgeons Ying Cen Jiake Chai Huade Chen Jian Chen Guanghua Guo Chunmao Han Dahai Hu Jingning Huan Xiaoyuan Huang Chiyu Jia Cecilia WP Li-Tsang Jianan Li Zongyu Li Qun Liu Yi Liu Gaoxing Luo Guozhong Lv Xihua Niu daizhi peng Yizhi peng Hongyan Qi Shunzhen Qi Zhiyong Sheng Dan Tang Yibing Wang Jun Wu Zhaofan Xia Weiguo Xie Hongming Yang Xianfeng Yi Lehua Yu Guoan Zhang The Chinese Burn Care and Rehabilitation Association Department of Burn and Plastic Surgery West China School of MedicineWest China HospitalSichuan UniversityChengduSichuanChina Department of Burn&Plastic Surgery the First Hospital Affiliated to General Hospital of PLABeijingChina Department of Burns General Hospital of Guangdong ProvinceGuangzhouGuangdongChina State Key Laboratory of Trauma Burns and Combined InjuryInstitute of Burn ResearchSouthwest Hospitalthe Third Military Medical UniversityChongqingChina Department of Burns the First Affiliated Hospital of Nanchang UniverisityResearch Center of Technology of Wound Repair Engineering in Jiangxi ProvinceNanchangJiangxiChina Department of Burns and Wound Center The Second Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhouZhejiangChina Department of Burns and Cutaneous Surgery Xijing HospitalFourth Military Medical UniversityXi’anShanxiChina Department of Burn and Plastic Surgery Ruijin HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghaiChina Department of Burns and Plastic Surgery Central South UniversityChangshaHunanChina Plastic Beauty and Burn Repair Center the 309th Hospital of the Chinese PLABeijingChina Department of Rehabilitation Sciences the Hong Kong Polytechnic UniversityHung HomHong KongChina Department of Rehabilitation Medicine the First Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsuChina Department of Burns and Plastic Surgery the Fifth Hospital of HarbinHarbinHeilongjiang ProvinceChina Department of Burn and Plastic Surgery the Fourth Hospital of TianjinBurn Institution of TianjinTianjinChina Burns and Plastic Surgery Center PLA Lanzhou General Hospital of Lanzhou CommandLanzhouGansuChina Department of Burn Surgery the Third People’s Hospital of WuxiJiangsuChina Department of Burn Surgery the First People’s Hospital of ZhengZhouZhengzhouHenanChina Department of Burn Surgery Beijing Children’s HospitalBeijingChina The Center of Burn and Plastic of Hebei Province Bethune International Peace Hospita

Quality of life and functional recovery after burn injury is the final goal of burn care,especially as most of burn patients survive the injury due to advanced medical ***,dysfunction,disfigurement,contractures,psychological problems and other discomforts due to burns and the consequent scars are common,and physical therapy and occupational therapy provide alternative treatments for these problems of burn *** guideline,organized by the Chinese Burn Association and Chinese Association of Burn Surgeons aims to emphasize the importance of team work in burn care and provide a brief introduction of the outlines of physical and occupational therapies during burn treatment,which is suitable for the current medical circumstances of *** can be used as the start of the tools for burn rehabilitation.

关键词： Burn Rehabilitation Physical therapy Occupational Therapy Scar

渗透冲击联合超声和去污剂处理对新生牛真皮组织学和生物活性成分的影响

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第三军医大学学报 2012年第9期34卷 797-800页

作者：陈博彭代智左海斌郑必祥周灵周新刘敬第三军医大学西南医院全军烧伤研究所创伤烧伤与复合伤国家重点实验室重庆400038

目的采用渗透冲击联合超声和去污剂处理新生牛真皮,观察处理前后新生牛真皮基质的组织学和生物活性成分的变化。方法从8头健康雄性新生牛皮肤获取网状层真皮组织。采用反复渗透压改变、超声震荡和去污剂联合法对新生牛网状层真皮进行处... 详细信息

目的采用渗透冲击联合超声和去污剂处理新生牛真皮,观察处理前后新生牛真皮基质的组织学和生物活性成分的变化。方法从8头健康雄性新生牛皮肤获取网状层真皮组织。采用反复渗透压改变、超声震荡和去污剂联合法对新生牛网状层真皮进行处理,HE染色、DAPI染色和核酸电泳检测细胞成分残留情况;扫描电镜(SEM)观察胶原束结构和细胞成分;PicoGreen法、DMMB法和BCA法对脱细胞前后真皮基质的DNA、sGAG和蛋白质含量进行定量分析;ELISA法测定脱细胞前后真皮基质内TGF-β1、EGF、bFGF和KGF含量。结果形态学观察可见脱细胞处理后的真皮三维结构完整,胶原纤维束排列较疏松,细胞清除较彻底,未见明显细胞及细胞碎片残留。与脱细胞处理前比较,脱细胞处理后的DNA含量[(2 516.1±324.2)ng/mg vs(249.5±53.8)ng/mg,P=0.000)]、sGAG含量[(5.92±0.50)μg/mg vs(2.48±0.24)μg/mg,P=0.000]、TGF-β1含量[(478.8±196.1)pg/g vs(180.3±111.0)pg/g,P=0.009]和EGF含量[(10.52±2.78)pg/g vs未能检测到EGF含量]均明显减少;而bFGF含量[(788.6±333.8)pg/g vs(364.8±294.8)pg/g,P=0.424]下降了53.7%,KGF含量[(0.033±0.000)pg/g vs(0.033±0.000)pg/g,P=0.433]变化不显著。结论反复渗透冲击联合超声和去污剂处理对新生牛真皮基质明显地清除了细胞及细胞碎片,较好地保留了真皮细胞外基质的三维结构和主要生物活性成分。

关键词：真皮细胞外基质渗透冲击超声生长因子

颗粒状脱细胞真皮基质的形态学特征及理化性能体外研究

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第三军医大学学报 2011年第1期33卷 4-8页

作者：左海斌彭代智郑必祥何升东周新刘敬第三军医大学西南医院全军烧伤研究所创伤烧伤与复合伤国家重点实验室重庆400038

目的研究颗粒状脱细胞真皮基质(particulate acellular dermal matrix,PADM)的体外形态学特征与理化性能。方法收集SD大鼠背部皮肤样本,采用本实验室的脱细胞方法制备片状脱细胞真皮基质(acellular dermal matrix,ADM)并分析其内胞核和... 详细信息

目的研究颗粒状脱细胞真皮基质(particulate acellular dermal matrix,PADM)的体外形态学特征与理化性能。方法收集SD大鼠背部皮肤样本,采用本实验室的脱细胞方法制备片状脱细胞真皮基质(acellular dermal matrix,ADM)并分析其内胞核和DNA残留情况。将片状ADM切割成不同规格的PADM,并结合大体观察、组织学分析、电镜技术、激光粒度分析仪和傅里叶变换红外光谱仪检测其外观轮廓、形态学、粒径分布和胶原分子结构。并将其复合人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)体外培养24 h,观察PADM与HUVEC的黏附情况。结果脱细胞方法有效降低了ADM的DNA含量[(1.51±0.37)μg/mgvs(0.30±0.09)μg/mg]。制备的PADM为白色颗粒,外形近似于长方体或立方体,显微镜下HE染色观察显示颗粒内部胶原纤维束结构疏松。扫描电镜显示颗粒断面结构疏松,透射电镜显示胶原原纤维周期性横纹结构和均匀分布的胶原原纤维间隙。激光粒度分析仪检测显示其粒度分布集中,例如,规格为0.2 mm的PADM,其80%(d0.1～d0.9)的粒径分布于233～487μm。FTIR结果显示,PADM分别在1 659、1 549、1 239 cm-1处出现的特征吸收峰,表明其保留了胶原分子的α-helix、β-sheet和β-turn二级结构,且体外HUVEC较易黏附于PADM。结论室温条件下,切割法制备的新型PADM形态规则,粒径分布集中,胶原纤维束形态和胶原分子的二级结构保存良好。

关键词：细胞外基质颗粒状脱细胞真皮基质粒径分布傅里叶变换红外光谱仪