四倍体滋养层干细胞的分离培养与鉴定

目的 建立可在体外培养的小鼠四倍体滋养层干细胞(TTSCs)系,为研究胎盘发育提供新的细胞模型.方法 用电融合的方法获得四倍体胚胎并进行体外培养,挑取克隆并传代后获得细胞系.通过中期染色体计数检测染色体数目;RT-qPCR、Western blot、免疫荧光检测标志基因表达;显微注射检测细胞的体内发育潜能.结果 建立了...     

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。     

烧伤后前臂骨筋膜室综合征的诊治

目的总结烧伤后前臂骨筋膜室综合征的诊治经验。方法回顾性分析近3年来我院收治的9例前臂烧伤后并发骨筋膜室综合征患者的临床资料。结果早期明确诊断6例,果断行筋膜切开减压治疗,上肢未出现功能障碍。3例在14～72小时后才明确诊断,虽经筋膜切开减压、脱水及改善微循环等处理但仍出现不同程度肌肉坏死,上肢遗留功能障碍。结论对于上肢深度烧伤,应早期行焦痂切开减压。上肢切、削痂后植皮创面,包扎不能过紧,术后前臂疼痛需考虑臂丛麻醉后可能减轻疼痛感等因素的影响,并结合观祭指端的感觉和运动功能,及早明确骨筋膜室综合征的诊断。     

氢氟酸烧伤合并重度吸入性损伤一例

患者男，28岁。因装盛氢氟酸（体积分数为55％）的容器爆裂致头面部、前胸及四肢烧伤。伤后即用冷水冲洗创面2～3min。1 h后收入笔者单位。入院时患者烦躁不安，体温37．2℃，脉搏89次／min，呼吸32次／min。双眼角膜混浊，口腔黏膜苍白，双肺呼吸音粗糙。头面部、左小腿、右大腿创面呈灰白色或皮革样，表面有散在斑点状黑色坏死灶，双前臂及前胸部创面基底苍白。     

非小细胞肺癌所致肺不张的放射治疗

临床上非小细胞肺癌 (ＮＳＣＬＣ)所致肺不张较为常见 ,常规放射治疗的疗效较低 ,近年由于采用非常规放射治疗取得了较好的疗效。我们自 1995年 1月至 1999年 12月 ,对ＮＳＣＬＣ所致肺不张采用大分次量放射治疗 ,获得较好的疗效 ,现报告如下。1　临床资料1.1　一般资料ＮＳＣＬＣ所致肺不张患者 14 8例 ,均经病理诊断及ＣＴ证实。男性 117例 ,女性 31例。年龄 43～ 74岁 ,中位年龄 5 7.8岁。肺鳞癌 114例 ,腺癌 2 5例 ,腺鳞癌 9例。ＡＪＣＣ分期 :Ⅱ期17例 ,Ⅲａ 期 10 4例 ,Ⅲｂ 期 2 7例。ＫＰＳ评分均≥ 6 0分。1.2　治疗方法①大分次量…     

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.     

应用时序序列探讨缺血性中风急性期证候与OCSP分型的相关性

目的使用时序序列探讨缺血性中风急性期证候与牛津郡社区卒中研究分型(OCSP)的相关性。方法选择846例缺血性中风急性期患者,进行OCSP分型,并将首次(发病1～3 d)、第5天、第7天、第10天、第14天作为证候演变的关键时间点,采用《缺血性中风证候要素诊断量表》判断证候,运用时序序列对4 230例次的临床数据进行分析。结果寻找出OCSP分型4种类型的核心证候经历5个时点的演变规律,得出40条规则。通过比较OCSP分型4种类型的首次、中间过程、最终的核心证候,发现存在的共性之处,及每一个OCSP分型证候演变的各自特征。结论缺血性中风急性期,中医证候与OCSP分型存在相关性,体现了"病证结合"的理念,为临床医师通过患者的中医证候把握疾病发展规律提供了数据支持。     

中枢型原始神经外胚层瘤一例

患者男，30岁。胸背部疼痛3个月余伴双下肢麻木半个月，MRI示T2～4椎体平面椎管内占位，于2001年7月6日在全麻下行T2～4椎管内硬膜下髓外肿瘤切除术，术后病理为髓内星形细胞瘤(Ⅱ级交界性)。术后3周行放疗，靶区包括T1～5，剂量44Gy分22次4．2周完成。放疗后胸背部疼痛及双下肢麻木明显减轻，每3个月复查MRI未见异常。2002年9月患者无明显诱因出现双下肢阵发性疼痛及触电感。MRI示T12～L1椎体平面椎管内占位，L5～S1椎管内见3个大小不一结节状块状信号。