正天丸治疗头痛临床研究

正天丸是首创于1985年的中成药复合组方,由钩藤、川芎、白芍、当归、地黄、白芷等15味中草药组成,具有疏风活血、养血平肝、通络止痛的功效,广泛适用于多种头痛.笔者将正天丸上市至今20余年有关临床研究总结分析如下.     

利用石膏颈部固定架制作颈部石蜡补偿块的技术应用

颈段食道癌由于颈部的活动度较大,颈部与肩部的组织厚度差异较大,因此其定位精度、摆位的重复性及射野内剂量的均匀性较差,使此部位肿瘤的放射治疗疗效受到一定的影响.虽然目前有颈肩部真空固定垫和颈肩部高分子塑料固定架解决了定位和摆位的精度,但未能解决颈肩部组织厚度差异较大的问题,因设备成本较高并非所有的放射治疗单位都有此装备.我科根据现有的条件用石膏绷带制作颈肩部固定架的同时利用石蜡作为等效补偿物对颈肩部的组织厚度进行补偿,在颈段食道癌的放射治疗中取得了较好的疗效现将制作方法报告如下.     

人骨髓间充质干细胞分离培养鉴定及VEGF基因的转染

目的：探讨人骨髓间充质干细胞（hMSC）分离培养、鉴定方法及人血管内皮生长因子（hVEGF）165基因转染hMSC后表达情况,拟构建血管化组织工程皮肤.方法：用密度梯度离心法分离骨髓间充质干细胞,筛选贴壁细胞培养并传代.观察细胞形态,生长情况.检验细胞CD44、CD29、CD14、CI45等表面标记.利用基因重组技术,构建pCDN3.1＋VEGF表达质粒,并将其转染第3代hMSC.经G418抗性筛选后扩大培养并用Western blot实验检测其蛋白产物.结果：骨髓密度梯度离心法分离hMSC,接种培养瓶后26 h呈对数生长,6d后达到高峰.hMSC表达CD29,CD44,不表达造血细胞标记物CD14和CD45.通过酶切鉴定验证成功构建pCDN3.1＋ VEGF表达质粒.Western blotting实验证实转染后hMSCs表达VEGF增加.结论：成功建立hMSC分离培养鉴定体系.利用基因重组技术可成功将hVEGF基因导入hMSC并产生预期效应.     

高压电击伤创面的修复

目的为了早期修复电击伤深部创面,避免并发严重感染、进行性坏死、深部组织外露和大出血,减少截肢率,最大限度地保持肢体的功能。方法早期创面彻底清创,祛除坏死病灶,有目的地保留虽变性而解剖结构尚完整的神经和肌腱,采用轴型带蒂皮瓣和肌皮瓣修复创面。结果58例皮瓣移植中仅4例皮瓣部分失活,1例皮瓣移植中断,需行截肢术,其余皮瓣成活良好,外形满意,肢体功能均有不同程度的改善。结论电击伤深部创面早期清创,应用皮瓣和肌皮瓣修复,可有效封闭创面,防止并发症,改善局部血运,保存肢体功能,降低截肢率。     

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RheA/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RheA,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RheA下游通路是增生性瘢痕治疗靶点之一.     

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.     

深圳某中专学生开展艾滋病知识健康教育效果评价

目的了解中专学生艾滋病相关知识知晓情况及其态度和行为，评价实施学校健康教育干预的效果。方法采用整群随机抽样方法，对深圳某中专学生于入学时和预防艾滋病健康教育后1个月，运用自编的同一问卷进行调查。结果教育前后，中专学生艾滋病知识知晓率有显著提高；对艾滋病相关的态度和行为，教育后有不同程度的改善；同伴教育是学生最愿意接受的教育方式；但在非传播途径和对性行为态度方面存在一定误区。结论对中专学生开展预防艾滋病健康教育有明显成效，艾滋病的非传播途径、性道德教育应作为艾滋病知识健康教育的重点内容；在今后的预防艾滋病健康教育应通过多种方式全方位开展，可重点采用同伴教育方式。     

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.     

积雪草苷对增生性瘢痕成纤维细胞结缔组织生长因子及RhoA/ROCK-Ⅰ调控信号的影响

 目的 研究积雪草苷对增生性瘢痕成纤维细胞结缔组织生长因子（CTGF）的影响及RhoA/ROCK-Ⅰ信号通路在其中的作用。方法 分离培养人增生性瘢痕组织来源的成纤维细胞,应用积雪草苷300 μg·mL-1及RhoA的下游效应器Rho激酶的特异抑制剂Y-27632对实验细胞进行干扰实验。分为对照组、积雪草苷组、Y-27632组、积雪草苷+ Y-27632组。运用荧光定量PCR（SYBR Green）及免疫荧光细胞化学的方法检测各组瘢痕成纤维细胞中RhA、ROCK-Ⅰ、CTGF mRNA与蛋白表达。结果 积雪草苷300 μg·mL-1能抑制RhoA、ROCK-Ⅰ、CTGF的mRNA与蛋白表达,与对照组比较具有统计学意义（P＜0.05）;Y-27632能阻碍积雪草苷的作用,积雪草苷（300 μg·mL-1）+ Y-27632组,CTGF的mRNA与蛋白表达明显高于单纯积雪草苷组（P＜0.05）,与对照组及Y-27632组比较无显著性差异（P＞0.05）;Y-27632组与对照组比较RhoA、ROCK-Ⅰ、CTGF的mRNA与蛋白表达没有统计学意义。结论 积雪草苷300 μg·mL-1能抑制RhoA、ROCK-Ⅰ、CTGF的mRNA与蛋白表达;而且可通过RhoA/ROCK-Ⅰ信号通路抑制瘢痕成纤维细胞中CTGF mRNA与蛋白表达。这可能是积雪草苷治疗瘢痕的作用机制之一。     

复方皂矾丸在放射治疗中的临床疗效观察

目的：观察复方皂矾丸加放疗治疗恶性肿瘤改善生存质量、升高白细胞作用与不良反应。方法：采用随机对照方法将120例患者分为复方皂矾丸加放疗的治疗组55例，单纯放疗的对照组65例。结果：治疗组升白有效率为74．5％，对照组为13．8％，两组有显著性差异（P〈0．01）。两组患者治疗前后症状改善，卡氏评分和体重变化均有显著差异（P〈0．05）。结论：复方皂矾丸可明显升高白细胞并改善生存质量，且不良反应小。