Regulation of PTC phenotype and function by TGF-β1: implications for transdifferentiation

Introduction There is now increasing evidence that proximal tubular cells (PTCs) contribute to renal interstitial fibrosis by alteration of matrix turnover and by the generation of pro‐fibrotic cytokines such as TGF‐β1. Recent studies suggest that, through a process of transdifferentiation, the PTCs are one source of the interstitial myofibroblasts that directly drive the fibrotic process. The aim of this work was to examine the role and mechanism by which TGF‐β1 may regulate PTC phenotype and function. Methods Experiments were performed using both primary‐cultures of PTC and the human PTC cell line HK2. All experiments were performed on growth‐arrested cells in the absence of serum. Results TGF‐β1 altered cell phenotype, assessed by light microscopy, with cells appearing elongated and spindle‐shaped. This was associated with loss of cell–cell contact and rearrangement of the actin cytoskeleton, increased formation of stress fibres and focal adhesions. Disruption of the actin cytoskeleton with cytochalasin‐D prevented phenotypic alterations following addition of TGF‐β1. Transient transfection with Smad‐2/‐4 or Smad‐3/‐4 expression vectors did not alter cell phenotype. Previously, we have demonstrated β‐catenin translocation to PTC nuclei and its association with Smad proteins following addition of TGF‐β1, suggesting the possibility that TGF‐β1 may modulate Wnt signalling. Wnt‐responsive Xtwn‐reporter construct was, however, silent in response to TGF‐β1. Similarly, a second Wnt‐/LEF‐1‐regulated element Toplflash, which does not contain Smad‐binding sites, was insensitive to TGF‐β1 signalling. In contrast, phenotypic changes in response to TGF‐β1 were abrogated by inhibitors of the RhoA downstream target ROCK, which also prevented loss of cell–cell contact and adherens junction disassembly. Removal of TGF‐β1 and addition of 1% FCS, however, reverted cell phenotype to a typical cobblestone epitheliod appearance, suggesting that TGF‐β1 did not result in terminal PTC transdifferentiation. Cells grown on tissue culture dishes coated with either type‐I or type‐III collagen also acquired an elongated fibroblastic phenotype; this effect was exaggerated by the addition of TGF‐β1. In contrast to the cells stimulated with TGF‐β1 alone, following stimulation by both TGF‐β1 and exposure to interstitial collagens, cell phenotype was stable in that it was not reversed upon removal of TGF‐β1 and addition of FCS. Addition of TGF‐β1 to cells grown on type‐IV collagen had no greater effect than TGF‐β1 alone. Addition of TGF‐β1 alone had little effect on the expression of α‐SMA. In contrast, cells grown on either type‐I or type‐III collagen, following addition of TGF‐β1, demonstrated marked increased expression of α‐SMA, which appeared to be incorporated into the cell cytoskeleton. Similarly, the combination of interstitial collagen (either type‐I or type‐III) and TGF‐β1 had synergistic effect on the relocation and down‐regulation of the epithelial markers E‐cadherin and cytokeratin. Finally, the results demonstrated synergistic effects of coating with interstitial collagen (either type‐I or type‐III), on cell ‘fibroblastic’ cell function as assessed by cell migration and by the synthesis of type‐III and type‐IV collagen. Conclusion The results of these in vitro experiments suggest that terminal transdifferentiation of proximal tubular epithelial cells is the result of a combination of the effects of the pro‐fibrotic cytokine TGF‐β1 and exposure of the cells to components of the interstitial extra‐cellular matrix to which the cells are not exposed in the absence of damage to the tubular basement membrane.     

CpG ODN增强乙肝疫苗在老年小鼠中的免疫应答

目的：探讨CpG ODN对老年小鼠的体液和细胞免疫应答的增强作用。方法：选用老年C57BL／6小鼠，将乙肝疫苗和10μg、20μg CpG ODN同时或单独肌注到小鼠体内，两周后以同样剂量加强免疫一次，再过3周后摘除眼球取血，用EILSA方法检测抗HBs16；抗体和IL-12；无菌取脾脏作HE染色，观察脾脏淋巴细胞变化。结果：10μg和20μg CpG ODN与疫苗同时注射组产生的抗体绝对量分别是单独注射疫苗组的3倍和4倍；产生的IL-12水平较对照组有明显升高，且20μg比10μCpG组产生的IL-12水平更高。光镜下各组的脾脏淋巴细胞的变化如下：正常老年鼠组脾脏淋巴细胞较正常青年鼠组明显稀少；老年鼠 10μgCpG组脾脏淋巴细胞较正常老年鼠组有了明显增加，且细胞核也明显增大；20μgCpG组增加的更加明显。结论：CpG ODN能增强乙肝疫苗在老年小鼠中的体液和细胞免疫应答。     

腰麻辅助硬膜外麻醉在全子宫切除术中的应用分析

目的:观察腰麻辅助硬膜外麻醉在全子宫切除术中的应用效果。方法:选择328例全子宫切除术病人,随机分为试验组(143例)、对照组(185例),均取L_(1-2)棘突间隙行硬膜外腔穿刺置管,对照组常规用药,试验组另取L_(3-4)棘突间隙行腰麻,观察各组的麻醉效果,术中探查牵拉子宫时病人的反应,血压及心率的变化。结果:试验组的麻醉良好率、肌松情况优于对照组(P<0.01),骶神经阻滞情况试验组完全阻滞,而对照组出现骶神经阻滞不全,首次剂量试验组少于对照组(P<0.01)。牵拉子宫后各组病人血压、心率均较牵拉前下降,且与牵拉前比较有显著差异(P<0.05)。结论:在全子宫切除术中应用腰麻辅助硬膜外麻醉优于单纯硬膜外麻醉,有一定临床应用意义。     

呼吸机相关肺炎的发生原因及护理对策探讨

目的　为进一步探讨发生呼吸机相关肺炎的因素和护理措施。方法　对 70例使用机械通气辅助治疗的重症监护病房 (ICU)病人 ,取深部痰液进行细菌培养与分离及药敏试验 ,并采用综合护理措施。结果　 70例患者除 4例因原发病病情恶化死亡外 ,无一例因呼吸机相关肺炎而导致病情恶化 ,延误治疗 ,且缩短了住院时间。结论　正确的气道管理 ,合理应用抗生素 ,良好的饮食护理等辅助治疗 ,能够降低呼吸机相关因素肺炎的发病率     

颞部-颌下联合进路切除咽旁颞下区巨大肿瘤(附2例报道)

[目的]探讨切除侵犯中颅窝的咽旁颞下区巨大肿瘤安全而彻底的手术进路.利用颞部切口可充分暴露中颅窝底和颞下窝以及颌下进路显露咽旁区解剖的特点,联合进路切除2例咽旁颞下区巨大肿瘤.2例侵犯中颅窝底的咽旁颞下区肿瘤顺利切除,患者术中组织损伤、出血量相对较少,术后无严重并发症.[结论]颞部-颌下联合进路适合于侵犯中颅窝底的咽旁颞下区肿瘤的手术切除.     

Protection of chickens against highly lethal H5N1 and H7N1 avian influenza viruses with a recombinant fowlpox virus co-expressing H5 haemagglutinin and N1 neuraminidase genes

Inactivated whole avian influenza virus (AIV) vaccine provides protection against homologous haemagglutinin (HA) subtype virus, but poor protection against a heterologous HA virus. Moreover, it induces chickens to produce antibodies to cross-reactive antigens, especially nucleoprotein, which is limits AIV serological surveillance. In this study, a recombinant fowlpox virus co-expressing HA (H5 subtype) and NA (N1 subtype) genes of AIV was evaluated for its ability to protect chickens against intramuscular challenge with a lethal dose of highly pathogenic (HP) AIV. Susceptible chickens were also vaccinated by wing-web puncture with the parent fowlpox vaccine virus. Following challenge 4 weeks later with HPAIV, all chickens vaccinated with recombinant virus were protected, while the chickens vaccinated with either the unaltered parent fowlpox vaccine virus or unvaccinated controls experienced 100% mortality following challenge. This protection was accompanied by the high levels of specific antibody to the respective components of the recombinant vaccine. The above results showed that rFPV-HA-NA could be a potential vaccine to replace current inactivated vaccines for preventing AI.     

经直肠超声引导前列腺穿刺活检203例临床分析

目的评估经直肠超声引导的前列腺六针穿刺活检在前列腺癌及前列腺其他疾病的诊断和鉴别诊断的价值。方法对指肛检查阳性，血清PSA〉4pg／L及经直肠超声检查前列腺声像图异常怀疑有占位性病变的203人进行经直肠超声引导的前列腺穿刺活检。结果穿刺活检的203例病理结果：良性前列腺增生（BPH）104例占51．24％，前列腺癌（PCa）95例占46．80％，前列腺结核及前列腺平滑肌肉瘤各2例，分别占0．98％。结论经直肠超声引导的前列腺穿刺活检其操作简单，病人痛苦小，并发症少，较安全。在前列腺癌及其他前列腺疾病的诊断与鉴别诊断中有重要的临床价值。     

茶氨酸对脑缺血再灌注损伤保护作用的实验研究

目的观察茶氨酸对脑缺血再灌注损伤的保护作用,为临床脑缺血再灌注损伤的预防和治疗提供实验依据。方法将24只健康家兔随机分为四组：假手术组、脑缺血组、茶氨酸予处理组和茶氨酸治疗组。麻醉后分离血管,采用基底动脉、双侧颈总动脉结扎法制备急性全脑缺血再灌注动物模型。假手术组仅分离动脉不结扎,予处理组在缺血前应用茶氨酸,治疗组在缺血后应用茶氨酸,缺血组不作特殊药物处理。各组分别在规定时间点即缺血前、再灌注后30min、1h、2h采血测定神经特异性烯醇化酶(NSE)含量,取脑组织活检观察超微结构,处死动物测定脑含水量。结果茶氨酸予处理组和治疗组以上各项指标均较脑缺血组有显著的改善,提示茶氨酸具有神经保护作用。实验还显示茶氨酸予处理组在再灌注30min时NSE含量与假手术组比较无差异,提示用茶氨酸予处理后可能使缺血后脑损害的发生延迟,为进一步的治疗争取了宝贵时间。结论茶氨酸对脑缺血再灌注损伤有保护作用,值得进一步研究。     

加强眶隔及重叠缝合眼轮匝肌在睑袋修复术中的应用

目的:研究应用加强眶隔及重叠缝合眼轮匝肌缘的方法治疗睑袋。方法:38例睑袋患者,应用加强眶隔及重叠缝合眼轮匝肌缘的手术方法去除睑袋,术后随访六个月。结果:术后27例得到1～6个月的随访,1例术后右眼外视时有牵扯感,2个月后消失。1例术后下睑皮下触及缝线结节,3个月后消失。其余均获得满意效果。结论:应用加强眶隔及重叠缝合眼轮匝肌缘的方法治疗睑袋,术后效果佳。     

辨证治疗特发性水肿43例疗效观察

特发性水肿是一种原因不明的综合征.在祖国医学属水肿.郁证范畴.近几年笔者采用辨证论治治疗本病43例,取得满意疗效,现报告如下.