戊二醛交联的异种/异体脱细胞真皮基质的制作及临床应用观察

目的：为降低异种／异体真皮组织的抗原性,便于临床大面积烧伤患者复合皮（ＣＳ）移植的推广。方法：将胰蛋白酶和戊二醛等处理的、含基底膜成分的１８例人（ｈ－ＣＳ）和８例猪（ｓ－ＣＳ）无细胞真皮基质（ＡＤＭ）与自体刃厚表皮一起重叠移植于１３种烧伤患者Ⅲ度切痂创面上,同时以邻近移植的自体刃厚皮（ＯＴＳ）和自体薄中厚皮（ＴＴＳ）为对照,观察无细胞真皮制作前后的组织结构变化和各组皮片移植的成活率及外观效果。结果     

降钙素基因相关肽对阿霉素心肌细胞毒性的保护作用

目的观察降钙素基因相关肽（ＣＧＲＰ）对阿霉素所致心肌细胞毒性作用的影响。方法采用原代培养乳鼠心肌细胞，以１０－６ｍｏｌＬ－１阿霉素造成急性心肌细胞毒性模型，ＣＧＲＰ作用浓度为１０－８ｍｏｌＬ－１。测定培养基中ＬＤＨ活性及心肌细胞内丙二醛（ＭＤＡ）、钙含量。透射电镜观察心肌细胞超微结构改变。结果阿霉素引起心肌细胞ＬＤＨ漏出明显增加；细胞内ＭＤＡ、钙含量水平明显升高；心肌细胞线粒体明显肿胀、嵴排列不整齐、断裂或消失，肌浆网明显扩张，染色质凝集。阿霉素所致心肌细胞ＬＤＨ漏出与细胞内ＭＤＡ含量水平呈正相关。ＣＧＲＰ可明显减轻阿霉素所致心肌细胞ＬＤＨ漏出及细胞内ＭＤＡ、钙的蓄积，明显减轻心肌细胞超微结构的改变。结论ＣＧＲＰ通过抑制脂质过氧化反应和减轻细胞内钙超载对阿霉素心肌细胞毒性具有保护作用     

复合皮移植的实验研究与临床应用

目的寻求一种较理想的真皮替代物,用以修复全厚皮肤缺损创面。方法(1)将异体／异种脱细胞真皮基质(allo／xeno-ADM) 自体刃厚皮组成复合皮(CS),以自体刃厚皮作对照,采用一步移植法进行实验研究和临床研究。移植术后应用生物化学及免疫学方法观察移植物抗原性的改变、表皮一真皮连接区(DEJ)基底膜带结构的再形成情况。(2)在此基础上,应用前述CS移植技术治疗53例全厚皮缺损或瘢痕切除患者,观察术后皮片成活情况并作随访。结果CS一步法移植后的成活情况良好。(1)移植后28周可见DEJ基底膜带重新形成,基底膜细胞呈极性生长并呈波浪形排列,真皮乳头与皮钉再生。allo-ADM的抗原性明显低于xeno-ADM,仅在术后早期有少数炎性细胞浸润,未见明显的免疫排斥反应。(2)移植于53例患者的70块CS中,65块完全存活占92．9％,2块因更换敷料不当部分存活,3块因创面感染移植失败。患者最长随访时间为8．5年,所移植的CS质地和色泽接近正常皮肤。结论allo／xeno-ADM的抗原性很低,移植后能发挥长期的真皮支架或诱导组织再生的真皮“模板”作用。     

抑癌基因MTS1对人瘢痕疙瘩成纤维细胞的生物学影响

目的：探讨多肿瘤抑制基因1（MTS1）对人瘢痕疙瘩成纤维细胞在生长增殖、DNA合成代谢及胶原合成量等方面的影响。方法：将外源性MTS1基因导入人瘢痕疙瘩成纤维细胞后，通过MTT法测定细胞的生长曲线，通过^3H-胸腺嘧啶核苷（^3H-TdR）及^3H-脯氨酸掺入的方法测定细胞的DNA合成量及胶原合成量，来分别比较转染前后细胞的生物学变化。结果：在转染MTS1后瘢痕疙瘩成纤维细胞的生长增殖受到明显的抑制，同时其DNA合成量及胶原合成量也明显降低，而转染空载体组及正常组细胞均未出现上述变化。结论：外源性多肿瘤抑制基因1(MTS1）能明显抑制人瘢痕疙瘩成纤维细胞在生长增殖、DNA合成及胶原合成等方面生物学功能，为瘢痕疙瘩的临床治疗提供了新的思路。     

ATP─MgCl2复合液抑制烫伤大鼠氧自由基和肿瘤坏死因子的生成

目的：研究ＡＴＰ－ＭｇＣｌ２对烧伤早期自由基及肿瘤坏死因子（ＴＮＦ）的生成所产生的影响．方法：通过大鼠３０％总体表面积（ＴＢＳＡ）三度烫伤模型观察经ＡＴＰ－ＭｇＣｌ２治疗后，血清ＴＮＦ－α生物学活性及全血细胞化学发光峰时和峰值的变化．结果：烫伤６ｈ后，血清ＴＮＦ－α生物学活性明显升高（Ｐ＜０．０５或Ｐ＜０．０１），至伤后２４ｈ均维持在较高水平．伤后立即经ＡＴＰ－ＭｇＣｌ２治疗可明显抑制血清ＴＮＦ－α活性升高．伤后６～２４ｈ，中性粒细胞（ＰＭＮ）产生氧自由基的能力明显增强（Ｐ＜０．０５或Ｐ＜０．０１），但血浆调理活性降低．伤后立即给予ＡＴＰ－ＭｇＣｌ２治疗，可抑制ＰＭＮ产生氧自由基的能力，同时提高血浆对ＰＭＮ的调理作用．结论：ＡＴＰ－ＭｇＣｌ２可降低烧伤早期炎症介质ＴＮＦ的生物学活性，抑制ＰＭＮ过度产生自由基，有利于严重烧伤机体重要脏器功能的维护     

离体角质形成细胞热损伤模型建立及细胞凋亡观察

Objective To reproduce a model of heat injured KC in vitro and explore its apoptosis rate of KC due to heat injury at different temperature. Methods Human KCs were cultured in vitro, and they were incubated at 37, 41, 43,45, 48, and 51 ℃ respectively for 10 mins in water bath. Trypan blue staining and Hoechst 33258 fluorescence staining were used respectively to determine necrosis and apoptosis of KC. Rates of apoptosis and necrosis of KC were analyzed quantitatively by flow cytometer. The prolifera-tion activity of KC after heat injury was detected by MTT test. Results The results of trypan blue stai-ning, Hoechst 33258 fluorescence staining, and flow cytometer demonstrated that number of apoptotic and necrotic KC increased gradually along with a rise of water bath temperature. The rates of apoptosis and necro-sis of KCwererespectively(12.3±3.2)% and (14.1±1.6)% at 45℃, (27.7±5.1)% and (58.0± 4.2)% at 48 ℃. Rate of KC necrosis reached up to (83.0±5.3)% at 51℃. Inhibition of KC growth reached a stationary phase when the injurious temperature reached 45 ℃ as observed with MTT test. Con-clusions Heat injury can induce apoptosis and growth inhibition of KC in vitro. Incubating KC at 45 ℃ for 10 mins is a good condition to reproduce a model of heat injured KC in vitro. This model may be used to study the biological character and apoptosis of KC after burn injury.     

ATP－MgCl_2对烫伤大鼠肠粘膜保护作用的实验观察

为研究ＡＴＰ－ＭｇＣｌ２对创伤应激动物肠粘膜的保护作用，采用３０％ＴＢＳＡⅢ度烫伤大鼠模型，观察了腹腔注射ＡＴＰ－ＭｇＣｌ２复合液对回肠粘膜丙二醛（ＭＤＡ）含量及回肠组织形态学改变的影响。结果显示：烫伤后６～２４小时，ＭＤＡ含量明显升高，并伴有形态学病理改变。经ＡＴＰ－ＭｇＣｌ２注射治疗的大鼠伤后２４小时内ＭＤＡ含量维持于正常对照水平，且回肠粘膜病理改变减轻。提示：ＡＴＰ－ＭｇＣｌ２能够保护烫伤大鼠肠粘膜，其作用机理可能与降低肠粘膜的脂质过氧化反应有关。     

离体角质形成细胞热损伤模型建立及细胞凋亡观察

目的 建立离体KC热损伤模型,了解不同温度热损伤后KC凋亡情况.方法 体外培养人KC,分别以37、41、43、45、48、51℃孵育KC 10 min.用锥虫蓝染色初步观察KC死亡情况,Hoechst 33258荧光染色检测细胞凋亡情况,应用流式细胞仪对细胞凋亡和死亡情况进行定量分析.采用噻唑蓝法检测热损伤对KC增殖活性的影响.结果 锥虫蓝染色、Hoechst 33258荧光染色、流式细胞仪检测结果 显示,随着热损伤温度的升高,KC中凋亡和死亡细胞逐渐增加;45℃时KC凋亡率和死亡率分别为(12.3±3.2)%、(14.1±1.6)%,48℃时各为(27.7±5.1)%、(58.0±4.2)%,51℃时KC死亡率高达(83.0±5.3)%.噻唑蓝法检测结果 显示,45℃热损伤条件下KC生长出现平台期.结论 热损伤能造成离体KC凋亡、生长受抑制,45℃孵育10 min为构建KC热损伤模型的较好条件.该模型可用于研究KC热损伤后的凋亡情况及其生物学特性.     

野生型p16基因体外诱导人瘢痕疙瘩成纤维细胞衰老的研究

目的：观察野生型p16基因对人瘢痕疙瘩成纤维细胞的生物学效应，探讨对瘢痕疙瘩进行基因治疗的可行性。方法：通过基因转染将含p16 cDNA的真核表达载体pcDNA3-p16导入人瘢痕疙瘩成纤维细胞中，筛选出阳性克隆后用测定细胞生长曲线、细胞周期及细胞超微结构等方法观察细胞的变化，用衰老相关-β-半乳糖苷酶染色法鉴定细胞形态。结果：细胞在转染野生型p16基因后自第3代起形态发生明显变化，细胞增殖明显减慢，细胞周期分析显示有96．7％的细胞处于G1期；电镜分析表明，细胞出现明显的溶酶体超载现象。衰老相关-β-半乳糖苷酶染色证实细胞进入衰老期。结论：野生型p16基因可诱导入瘢痕疙瘩成纤维细胞提前进入衰老阶段，这为瘢痕疙瘩基因治疗提供了实验依据。     

离体角质形成细胞热损伤模型建立及细胞凋亡观察

Objective To reproduce a model of heat injured KC in vitro and explore its apoptosis rate of KC due to heat injury at different temperature. Methods Human KCs were cultured in vitro, and they were incubated at 37, 41, 43,45, 48, and 51 ℃ respectively for 10 mins in water bath. Trypan blue staining and Hoechst 33258 fluorescence staining were used respectively to determine necrosis and apoptosis of KC. Rates of apoptosis and necrosis of KC were analyzed quantitatively by flow cytometer. The prolifera-tion activity of KC after heat injury was detected by MTT test. Results The results of trypan blue stai-ning, Hoechst 33258 fluorescence staining, and flow cytometer demonstrated that number of apoptotic and necrotic KC increased gradually along with a rise of water bath temperature. The rates of apoptosis and necro-sis of KCwererespectively(12.3±3.2)% and (14.1±1.6)% at 45℃, (27.7±5.1)% and (58.0± 4.2)% at 48 ℃. Rate of KC necrosis reached up to (83.0±5.3)% at 51℃. Inhibition of KC growth reached a stationary phase when the injurious temperature reached 45 ℃ as observed with MTT test. Con-clusions Heat injury can induce apoptosis and growth inhibition of KC in vitro. Incubating KC at 45 ℃ for 10 mins is a good condition to reproduce a model of heat injured KC in vitro. This model may be used to study the biological character and apoptosis of KC after burn injury.