几丁质膜作为表皮干细胞培养支架的实验研究

Objective To investigate the feasibility of constructing a skin tissue engineering cover-ing on chitinous membrane using rat epidermal stem cells (ESCs). Methods Rat ESCs were isolated and cultured by cold digestive method and collagen type Ⅳ adherent method. Cell colonies were observed with in-verted microscope. Expressions of DNA and RNA of ESCs were detected with laser scanning confocal micro-scope. Growth curves of cells were determined with Alamar BlueTM colorimetric method. Expressions of sur-face markers of ESCs (CD29, CD71, CD49d, and CD34) were detected with flow cytometer. Positive expres-sions of CK15, CK19, and P63 of ESCs were determined by immunohistochemistry. Influence of original chi-tinous membrane leachate in different dilutions on ESCs was observed. Condition of growth of ESCs on the ve-hicle was observed. Results Isolated cultured cells were verified as ESCs, of which the doubling generation time was 48 hs. CD29 and CD49d were positive;CD71 and CD34 were negative;CKi9, CK15, and P63 were positive. Compared with that of control group, ESCs cultured in chitinous membrane leachate showed slight cell proliferation when diluted to 1:8-1:512 dilutions, but there was no statistically significant difference (P>0.05). The checkerboard-form cell colonies of ESCs could be visualized with naked eyes on the chi-tinous membrane in 2-4 weeks of culture. A multitude of ESCs were seen to grow on fibres under microscope. Conclusions Chitinous membrane may be used as ESCs culture vehicle, and biological compatibility is good.     

中风病证候标准发展历程述评

本文分析“中风病中医诊断、疗效评定标准”(一代标准)、“脑血管病中医辨证量表”、“中风病辨证诊断标准”、“中风病诊断与疗效评定标准”(二代标准)、“缺血性中风证候要素诊断量表”的重要意义和不足之处,并对证候诊断标准的研制进行方法学探讨,体现了证候诊断、疗效评价的标准化对中医药诊疗体系的重要性。     

和肝颗粒治疗肝郁脾虚型非糜烂性反流病疗效及对5-HT,VIP的影响

目的:观察和肝颗粒治疗肝郁脾虚型非糜烂性反流病患者的临床疗效及对患者生活质量,5-羟色胺(5-HT),血管活性肠肽(VIP)水平的影响。方法:将肝郁脾虚型非糜烂性反流病患者随机分为治疗组32例和对照组33例,治疗组采用和肝颗粒10 g/袋,每次1袋,每日2次,餐后口服。对照组采用奥美拉唑肠溶片20 mg/粒,每日2次,餐前口服。两组疗程均为8周。观察两组治疗前后中医症状总积分及SF-36健康量表评分及5-HT,VIP水平。结果:两组患者中医疗效比较,治疗组、对照组中医疗效总有效率分别为87. 5%,81. 8%,治疗组优于对照组(P 0. 05);治疗后治疗组在特殊症状、次要症状方面改善优于对照组(P 0. 05);在主要症状方面两组疗效相当;治疗后治疗组SF-36健康量表在生理职能、总体健康、活力、社会功能、情感职能、心理健康6个维度评分均较高于对照组(P 0. 05);治疗后两组患者5-HT,VIP水平均降低,治疗组低于对照组(P 0. 05)。结论:和肝颗粒治疗肝郁脾虚型非糜烂性反流病患者具有较好的临床疗效,能够改善患者的中医症状,并能提高患者的生活质量,降低食管内脏敏感性。     

《缺血性中风证候要素诊断量表》临床验证

目的 验证《缺血性中风病证候诊断量表》的科学性和可操作性.方法 采用中医证候诊断量表4+1的临床验证新模式,进行了两轮多中心临床验证,共有21个省市的23家脑病专科协作组单位参加,共验证急性期和恢复期不同时点的中风患者1 053例.结果 《缺血性中风证候要素诊断量表》在特异性、敏感性和判断准确率上明显优于1994年《中风病辨证诊断标准》.结论 该量表适合于对中风病急性期和恢复早期患者证候的诊断,可操作性强.     

深圳市龙岗区部分中专学生营养知识、态度及饮食行为调查

目的 了解深圳市龙岗中专学生的营养知识、态度和饮食行为,为开展营养健康教育及干预提供依据.方法 采用自行设计的营养知识、态度及饮食行为问卷,对龙岗中专278名学生进行营养知识、态度及饮食行为调查.结果 学生对营养知识缺乏全面深入的了解,营养知识知晓率为51.3%;普遍存在不能坚持每天吃早餐,三餐食量分配不合理等不良饮食行为.结论 应采取多种形式对龙岗中专学生加强营养知识的健康教育和饮食行为的正确引导.     

几丁质膜作为表皮干细胞培养支架的实验研究

Objective To investigate the feasibility of constructing a skin tissue engineering cover-ing on chitinous membrane using rat epidermal stem cells (ESCs). Methods Rat ESCs were isolated and cultured by cold digestive method and collagen type Ⅳ adherent method. Cell colonies were observed with in-verted microscope. Expressions of DNA and RNA of ESCs were detected with laser scanning confocal micro-scope. Growth curves of cells were determined with Alamar BlueTM colorimetric method. Expressions of sur-face markers of ESCs (CD29, CD71, CD49d, and CD34) were detected with flow cytometer. Positive expres-sions of CK15, CK19, and P63 of ESCs were determined by immunohistochemistry. Influence of original chi-tinous membrane leachate in different dilutions on ESCs was observed. Condition of growth of ESCs on the ve-hicle was observed. Results Isolated cultured cells were verified as ESCs, of which the doubling generation time was 48 hs. CD29 and CD49d were positive;CD71 and CD34 were negative;CKi9, CK15, and P63 were positive. Compared with that of control group, ESCs cultured in chitinous membrane leachate showed slight cell proliferation when diluted to 1:8-1:512 dilutions, but there was no statistically significant difference (P>0.05). The checkerboard-form cell colonies of ESCs could be visualized with naked eyes on the chi-tinous membrane in 2-4 weeks of culture. A multitude of ESCs were seen to grow on fibres under microscope. Conclusions Chitinous membrane may be used as ESCs culture vehicle, and biological compatibility is good.     

乳没生肌散对组织工程皮肤修复皮肤缺损的研究

目的 研究乳没生肌散对组织工程皮肤修复皮肤缺损创面的促进作用.方法 将人成纤维细胞种植入胶原海绵,人角质形成细胞种植在表层,构建组织工程皮肤,并移植入裸鼠皮肤缺损创面.移植后2 d,将SPF级裸鼠随机分为实验组和模型对照组,每组4只.实验组选用乳没生肌散对创面进行外敷治疗,隔日换药;模型对照组创面不用药物治疗,隔日涂以生理盐水.组织工程皮肤移植后9 d取材,取移植处组织行冰冻切片,HE染色,观察创面局部表皮厚度及真皮中毛细血管增生情况,免疫荧光双标检测真皮层中Ln、Ⅰ型胶原的表达.结果 组织工程皮肤移植后9 d,实验组与模型对照组相比,创面移植处表皮增厚,真皮中毛细血管增生较多,Ln、Ⅰ型胶原的的分泌增强.结论 乳没生肌散具有促进组织工程皮肤修复皮肤创面的作用.     

脱模——带模法制作金属烤瓷长桥支架

金属烤瓷桥是目前口腔修复中应用较广泛的一种修复体 ,深受患者欢迎。但金属烤瓷长桥支架由于其跨度长、容易变形 ,在制作中存在较大的难度。在常规制作此类支架的方法中 ,通常有带模铸造法、脱模铸造法、分段制作焊接法。笔者结合带模铸造法和脱模铸造的优点 ,总结出脱模—带模铸造法制作金属烤瓷长桥支架 ,效果较为理想。　　作者单位 :5 10 0 60广州 ,中山医科大学光华口腔医院　　 1.材料 :嵌体蜡、间隙剂、磷酸盐系冠桥包埋料、观测仪、Ｎｉ—Ｃｒ合金、琼脂、真空搅拌机、高频铸造机。2 .方法 :①石膏模型处理 ,用石膏打磨机修整好石…     

RhoA/ROCK-I信号通路与增生性瘢痕成纤维细胞结缔组织生长因子的表达

背景：前期实验表明增生性瘢痕中RhoA和ROCK-I基因表达较正常皮肤高,提示RhoA/ROCK-I信号通路可能参与了增生性瘢痕的发生,但其在病理性瘢痕中的作用尚不清楚。 目的：研究RhoA/ROCK-I信号通路在增生性瘢痕成纤维细胞结缔组织生长因子(connective tissue growth factor,CTGF)表达调控中的作用。 方法：分离培养人增生性瘢痕组织来源的成纤维细胞,应用转化生长因子β1及Rho激酶的特异抑制剂Y-27632对细胞进行干预实验。采用实时荧光定量PCR及免疫荧光细胞化学方法检测瘢痕成纤维细胞中RhoA,ROCK-I及CTGF mRNA与蛋白的表达。 结果与结论：给予转化生长因子β1后,增生性瘢痕成纤维细胞中RhoA,ROCK-I及CTGF mRNA与蛋白表达明显增多(P < 0.01);而Y-27632能阻碍转化生长因子β1的作用;但单独给予Y-27632并不引起瘢痕成纤维细胞中RhoA,ROCK-I及CTGF的mRNA与蛋白表达改变。说明转化生长因子β1可通过RhoA/ROCK-I信号通路调控CTGF mRNA与蛋白的表达,即RhoA/ROCK-I信号通路参与了瘢痕成纤维细胞CTGF的表达调控,阻断RhoA下游通路是增生性瘢痕治疗靶点之一。     

17例心脏刀刺伤的急救体会

目的：分析本院1990年10月－2000年10月17例心脏刀刺伤患者的抢救经过，以提高早期 诊断和急救水平。方法：根据外伤史，临床表现，必要的体表伤口扩大探查及其他辅助检查确定诊断，17例患者剖胸探查。结果：抢救成功14例,死亡3例，抢救 成功率为82.4%。其中3例患者抢救无效导致死亡的原因都和未能及时明确诊断，果料剖胸探查有关。结论；心脏刀刺伤抢救的 关键在于 及时确明诊断，抗休克的同时果断剖胸解除心包压塞，修补心肌裂口。