Assessment of oral ciprofloxacin impaired gut barrier integrity on gut bacteria in mice

Gut bacteria and gut barrier plays important roles in body homeostasis. Ciprofloxacin (CPFX) is widely used to treat bacterial infections. However, whether high dosage of CPFX has side effects on gut barrier integrity is still unclear. Our results indicated that the High CPFX treatment (1 mg/ml) caused weight loss, nervousness, anorexia, and increased apoptosis cells in gut, but less influence was observed in the Low CPFX group (0.2 mg/ml). Meanwhile, the High CPFX treatment impaired tight junction molecules Ocln/ZO-1 level and down-regulated antibacterial genes expression (reg3γ, pla2g2α and defb1). Further, the High CPFX treatment increased pro-inflammatory cytokine IL-1β in intestinal tract, decreased IL-17A of duodenum but increased IL-17A of colon at day 37. In addition, the gut bacterial diversity and richness behaved significantly loss regarding CPFX treatment, especially in the High CPFX group during the experiment. Indole exhibited sharply decline in both Low and High CPFX groups at day 7, and the High CPFX mice needed longer time on restoring indole level. Meanwhile, CPFX treatment strongly decreased the concentrations of butyric acid and valeric acid at day 1. Correlation analysis indicated that the linked patterns between the key bacteria (families Bacteroidales_S247, Ruminococcaceae and Desulfovibrionaceae) and metabolites (indole and butyric acid) were disturbed via the CPFX treatment. In conclusion, the High CPFX treatment impaired the gut barrier with the evidence of reduced expression of tight junction proteins, increased apoptosis cells and inflammatory cells, decreased the bacterial diversity and composition, which suggesting a proper antibiotic-dosage use should be carefully considered in disease treatment.     

阴道粘膜上皮细胞的分离培养和鉴定

目的 初步建立阴道粘膜上皮细胞体外培养方法,为阴道粘膜上皮研究提供实验模型.方法 取雌性新西兰大白兔阴道粘膜组织小块,胶原酶Ⅳ和胰蛋白酶联合消化分离法收集上皮细胞,接种于角朊细胞无血清培养液中静置培养、传代.动态观察细胞生长增殖情况,扫描和透射电镜观察超微结构,流式细胞仪测定细胞增殖周期,并进行免疫组织化学鉴定.结果 体外培养的阴道粘膜上皮细胞为二倍体细胞,增殖状态良好,细胞间可见桥粒连接,免疫组化角蛋白染色阳性.细胞的超微结构和免疫组化染色均具有上皮细胞特征.结论 在本实验条件下,体外培养的阴道粘膜上皮细胞具有较好的增殖能力,可作为阴道粘膜上皮研究的理想实验模型.     

CFTR stabilizes ENaC at the plasma membrane

CFTR was reported to regulate ENaC channel opening, decreasing ENaC activity in airways and increasing it in sweat ducts. We generated MDCK-I cell lines stably expressing tagged alphabetagammaENaC+CFTR or ENaC alone, and developed an assay to quantify cell-surface half-life of ENaC. Surprisingly, we found that co-expressed CFTR stabilizes ENaC at the plasma membrane, suggesting that CFTR regulates ENaC stability, not just opening.     

外源性三磷酸腺苷对肾小管上皮细胞增殖的影响

观察外源性三磷酸腺苷（ＡＴＰ）对肾小管上皮细胞株ＬＬＣＰＫ１增殖的影响。方法通过测定３Ｈ－胸腺嘧啶掺入、细胞计数以及细胞内丝裂素活化蛋白激酶活性，并与表皮生长因子（ＥＧＦ）的作用比较。结果发现ＡＴＰ呈浓度依赖性促进细胞的ＤＮＡ合成，并可使细胞计数增加，同时激活细胞内的丝裂素活化蛋白激酶，其作用与ＥＧＦ相似。腺苷与ＡＴＰ有类似作用，但较弱。核苷酸转运蛋白抑制剂对４－硝基苯６－硫基甙并不能抑制ＡＴＰ及腺苷的作用。结论细胞外ＡＴＰ可促进肾小管上皮细胞增殖，并可增强ＥＧＦ的促增殖作用，这一作用可能是通过膜受体介导的细胞内丝裂素活化蛋白激酶活化而实现的     

病理性损伤因素对肾小管上皮细胞表型转化的影响

目的 :观察低血清、高糖、高蛋白刺激下肾小管上皮细胞向间叶性细胞的表型转化 ,探讨损伤的肾小管上皮细胞在肾小管间质纤维化中的作用。方法 :以体外培养的人近端肾小管上皮细胞为对象 ,分别用含低血清(0 .2mol·L-1小牛血清 )、高葡萄糖 (4.5 g·L-1)、高白蛋白 (15 g·L-1)的培养液培养 7d ,透射电镜检测细胞形态变化 ;免疫组化和Westernblot检测细胞表型改变 ,包括CK、Vimentin、α SMA、Ⅰ和Ⅲ型胶原 ,原位杂交检测Ⅰ型胶原mRNA表达 ,同时检测细胞TGFβ1蛋白表达。 结果 :低血清、高糖、高蛋白直接刺激下 ,肾小管上皮细胞出现明显形态学改变 ,包括细胞变为长梭形 ;透射电镜下细胞内线粒体明显减少 ,粗面内质网明显增加 ,并出现actin样微丝。免疫组化染色和Westernblot显示CK表达明显减弱 ,Vimentin、Ⅰ、Ⅲ型胶原、α SMA和TGFβ1表达增强。原位杂交显示Ⅰ型胶原mRNA表达增强。结论 :低血清、高糖、高蛋白可直接引起人近端肾小管上皮细胞TGFβ1表达增加 ,并发生向间叶性细胞的表型转化     

毛囊细胞移植诱导裸鼠毛囊样结构形成的研究

目的:观察毛囊细胞移植诱导裸鼠毛发再生和毛囊重建情况.方法:采用细胞培养技术和裸鼠移植技术,将培养的毛囊毛乳头细胞、真皮鞘细胞和头皮真皮成纤维细胞按比例与毛囊上皮细胞混合,移植到裸鼠皮下,观察毛囊形成情况.结果:在毛囊毛乳头细胞与毛囊上皮细胞混合后移植到裸鼠皮下后可见毛囊样结构形成,而毛囊真皮鞘细胞和头皮成纤维细胞与毛囊上皮细胞混合则不能诱导裸鼠毛囊样结构形成.结论:低传代培养的毛乳头细胞与毛囊上皮细胞混合后在体内可诱导毛囊样结构形成.     

The estimation of mean nuclear volume in the diagnosis of breast carcinoma.

Sixty-two cases of breast pathology were randomly selected from the files of the Dunedin Public Hospital for evaluation of mean epithelial nuclear volume. The cases were comprised of both benign and malignant ductal epithelial disease, diagnosed in cytological smears or in histological sections. Nuclear volume in histological preparations was estimated by the stereological technique of point-line intercept measurements to derive volume-weighted mean volumes (vV). An index of the nuclear volume (Vi) of cytology smears was calculated from measurements of nuclear areas by either image analysis Vi(e) or point-line intercepts Vi(p). By all methods of analysis a clear distinction of nuclear volume was found between the benign [means for the cytology were 148 microns 3 (Vi(e)) or 246 microns 3 (Vi(p)), and 203 microns 3 (Vv) for the histology specimens] and malignant diseased cases [means for cytology: 524 microns 3 (Vi(e)) or 886 microns 3 (Vi(p)), and 587 microns 3 (vV) for the histology specimens].     

喉上皮细胞条件液诱导胚胎干细胞分化为上皮细胞的实验研究

目的:探讨体外用喉上皮细胞条件培养液诱导胚胎干细胞分化为上皮细胞的方法及诱导条件。方法:将E14小鼠胚胎干细胞体外培养形成拟胚体后，在人喉上皮细胞条件培养液及因子诱导条件下共培养 14 d，形态学观察并采用免疫细胞化学方法检测上皮细胞的角蛋白Ck 4、14、19蛋白质水平的表达情况。结果:在诱导14 d的培养系统中较对照细胞可见更多上皮样细胞出现，免疫细胞化学染色显示上皮样细胞胞浆中可见角蛋白Ck 4、14、19均有表达。结论:胚胎干细胞在喉上皮细胞条件培养液诱导条件下可形成上皮样细胞，为组织工程制作人工喉上皮材料奠定了实验基础。