bFGF和硫糖铝局部应用促扩张术方法研究

目的探讨碱性成纤维细胞生长因子(bFGF)不同剂量、用药方法对扩张组织的影响.方法以白色家猪为实验动物,采用持续恒压扩张技术,观察不同剂量和不同给药时间,bFGF和硫糖铝对扩张组织的影响及组织结构变化.结果扩张术同时每日2次局部应用bFGF 9AU/cm2 硫糖铝100μg/ml,共7 d,浸于明胶海绵上缓慢持续释放对扩张组织的影响最明显,效果最佳,扩张器内实际注液量、扩张所获得的皮肤净增面积明显增加,皮瓣回缩率降低.表皮细胞层数增多,胶原纤维、弹力纤维、成纤维细胞密度和毛细血管密度显著增高.药量增加效果未随之增加.结论持续恒压扩张术时合用bF-GF 9AU/cm2 硫糖铝100μg/ml,每日2次注药浸入明胶海绵持续缓慢释放效果最佳.     

碱性成纤维细胞生长因子在实验性硬脑膜重建中的应用

[目的] 探讨碱性成纤维细胞生长因子( bFGF)在实验性自体筋膜硬脑膜重建中的应用价值.[方法]取 SD大鼠 35只,分成 5组.其中 A组 3只,直接将背部筋膜覆盖在硬膜外, B组 5只,采用自体筋膜硬脑膜重建模型,通过免疫组织化学方法观察这两组大鼠自体筋膜硬脑膜移植的愈合过程及细胞因子 bFGF在其中的表达; C组、 D组、 E组各 9只,分别进行自体筋膜硬脑膜重建, D组加用外源性 bFGF, E组仅用明胶海绵,通过脑脊液漏研究、免疫组织化学方法研究外源性 bFGF对自体筋膜重建硬脑膜愈合过程的影响并用 RT- PCR方法研究内源性 bFGF mRNA的表达情况.[结果] 大鼠自体筋膜移植硬脑膜部位 bFGF表达明显; C、 D、 E组大鼠重建硬脑膜抵抗脑脊液漏压力值( mmH2O)分别为 311± 75, 497± 153, 338± 88,Ⅰ型胶原纤维表达值分别为 4.9± 0.8, 10.9± 1.6, 5.3± 0.9, D组均好于对照组( P< 0.05);内源性 bFGF mRNA的表达没有明显变化.[结论] 在自体筋膜移植硬脑膜的愈合过程中 bFGF可能起了重要的作用;外源性 bFGF处理的大鼠其移植筋膜愈合要好于对照组.     

Relaxin down-regulates renal fibroblast function and promotes matrix remodelling in vitro.

BACKGROUND: Renal fibroblasts are important effector cells in tubulointerstitial fibrosis, with experimental antifibrotic strategies focusing on the functional down-regulation of these cells. Several experimental models of fibrosis have provided evidence for the effectiveness of the polypeptide hormone relaxin as a potential antifibrotic agent. This study was conducted to further elucidate the antifibrotic mechanisms of relaxin on renal fibroblasts in vitro. METHODS: Rat cortical fibroblasts were obtained from outgrowth culture of renal tissue isolated from kidneys 3 days post-unilateral ureteric obstruction and constituted 100% of cells studied. A relaxin radio-receptor assay was used to establish binding of relaxin to renal fibroblasts in vitro. Functional studies then examined the effects of H2 relaxin (0, 1, 10 and 100 ng/ml) on fibroblast kinetics, expression of alpha-smooth muscle actin (alpha-SMA), total collagen synthesis, collagenase production and collagen-I lattice contraction. CTGF mRNA expression was also measured by northern analysis. RESULTS: H2 relaxin bound with high affinity to rat renal fibroblasts, but receptor numbers were low. Consistent with its previously reported bimodal effect, transforming growth factor (TGF-beta 1) reduced fibroblast proliferation, an effect abrogated by H2 relaxin. Fibroblasts exposed to H2 relaxin (100 ng/ml) for 24 h demonstrated decreased immunostaining for alpha-SMA and reduced alpha-SMA protein expression compared with controls. There was a trend for a relaxin-mediated reduction in total collagen synthesis and alpha 1(I) mRNA expression with large dose-related increases in collagenase protein expression being observed. TGF-beta 1-stimulated collagen-I lattice contraction was significantly inhibited following co-incubation with 100 ng/ml relaxin. Incremental doses of H2 relaxin had no significant effect on CTGF mRNA expression. CONCLUSIONS: The findings of this study suggest that the antifibrotic effects of relaxin involve down-regulation of fibroblast activity, increase in collagenase synthesis and restructuring of collagen-I lattices, which are consistent with its known physiological role of matrix remodelling. Although there appears to be an interaction between TGF-beta 1 and H2 relaxin, this does not appear to involve a reduction in CTGF mRNA expression.     

转移因子口服液佐治小儿单纯性肾病综合征临床研究

目的：观察转移因子口服液佐治小儿单纯性肾病综合征的疗效。方法：将75例诊断为单纯性肾病综合征的患儿随机分为治疗组和对照组,对照组主要采用泼尼松口服治疗,治疗组另加用转移因子口服液治疗。观察两组的感染率、复发率及治疗前后血IgG、IgA水平。结果：治疗组的感染率和复发率明显较低;治疗后血IgG、IgA水平明显提高,与对照组比较差异明显（P〈0.01）。结论：转移因子口服液佐治小儿单纯性肾病综合征可以降低患者的感染率和复发率,提高血IgG和IgA水平,是治疗肾病综合征的有效方法之一。     

Thyrotrophin-Releasing Hormone Raises Cytosolic Free Calcium Concentration in Human Adenomatous Somatotrophs and Corticotrophs; Comparison with in vivo Responsiveness to Thyrotrophin-Releasing Hormone in Patients with Acromegaly or Cushing's Disease

The effect of thyrotrophin-releasing hormone (TRH) on intracellular free Ca²⁺ concentration, [Ca²⁺)i, was investigated with the fluorescent dye fura-2 in cell suspensions obtained from 13 human growth hormone-secreting adenomas and 6 adrenocorticotrophin-secreting adenomas. Preoperatively, 9 out of 13 acromegalic patients showed a positive growth hormone response to TRH administration while none of the 6 patients with Cushing's disease had a plasma adrenocorticotrophin increase after TRH injection. In all the growth hormone-secreting adenomas the addition of TRH (100 nM) caused a significant rise in [Ca²⁺]i (from a resting level of 133±40 (±SD) to a value of 284±119 nM at 100 nM TRH, n = 42; P<0.001). The transient induced by TRH was found to have a dual origin, one due to Ca²⁺ mobilization from intracellular stores which was maintained in presence of EGTA (3mM) and verapamil (10 μM) and a plateau phase due to Ca²⁺ influx from the extracellular media. Somatostatin (0.1 μM) lowered both resting [Ca²⁺]i and TRH-induced transients. The effect of gonadotrophin-releasing hormone on [Ca²⁺]i was evaluated on cell suspensions obtained from 6 growth hormone-secreting adenomas. Gonadotrophin-releasing hormone (100 nM) caused a marked rise in [Ca²⁺]i (from 179±25 to 283±15nM) on the cell suspension obtained from the only in vivo responsive adenoma while it was ineffective in the remaining 5. Although TRH was ineffective in modifying plasma adrenocorticotrophin levels in all patients with Cushing's disease, in 5 out of 6 tumors the addition of 100 nM TRH caused a significant rise in [Ca²⁺]i (from 102.5 ± 36 to 163±66 nM, n = 22; P < 0.005). However, the effect of TRH on [Ca²⁺]i was significantly lower than that caused by arginine vasopressin, a physiological stimulator of adrenocorticotrophin release ([Ca²⁺]i values; 145±78 nM at 100 nM TRH versus 300±140 at 10 nM arginine vasopressin, n = 15; P<0.05). Moreover, the effect of arginine vasopressin on [Ca²⁺]i was detectable at concentrations as low as 0.1 nM while TRH was effective at concentrations higher than 1 nM. By contrast, gonadotrophin-releasing hormone was ineffective in increasing [Ca²]i in all the adrenocorticotrophin-secreting adenomas studied. Collectively, these data indicate that sensitivity to TRH is present in almost all the growth hormone- and adrenocorticotrophin-secreting adenomas independently of the responsiveness of the individual patients to the peptide.     

沙培林增强人腹腔巨噬细胞抗癌免疫功能的研究

目的:探讨腹腔内注射沙培林增强人腹腔抗癌免疫功能的机制。方法:72例早中期胃肠道肿瘤患者术前48h和24h腹腔内分别注射生理盐水和5KE的沙培林,术中采集腹腔内巨噬细胞,计数并测定乳酸脱氢酶(LDH)和酸性磷酸酶(ACP)的活性,巨噬细胞吞噬活力,一氧化氮(NO)的分泌以及对人胃癌MKN1细胞的细胞毒性进行分析。同时采集大网膜,对大网膜乳斑的数量和面积进行观察。结果:沙培林显著增加腹腔巨噬细胞(PMΦ)的数量和NO的分泌,增强LDH和ACP的活性,吞噬活力,以及抗癌细胞毒性,也显著增加了大网膜乳斑的数量和面积。结论:腹腔内注射沙培林可显著增加人大网膜乳斑的数量和面积,并因此增加PMΦ的数量,增强PMΦ的活性。因而增强了腹腔巨噬细胞的免疫功能。     

AdEasy1／Cbfa1诱导间充质干细胞向成骨细胞分化的实验研究

目的：观察核心结合因子a1（Cbfa1）对兔骨髓间充质干细胞（MSCs）向成骨细胞分化的诱导作用。方法：体外分离培养兔骨髓MSCs，用AdEasy1／Cbfa1。转染MSCs，在转染后3d,1、2、3和4周时，组织化学和免疫组化等方法检测成骨标志碱性磷酸酶和骨钙素的表达。结果：AdEasy1／Cbfa1转染后的兔骨髓MSCs表现出与成骨细胞相似的形态，并且表达碱性磷酸酶和骨钙素。结论：Cbfa1可诱导兔骨髓MSCs向成骨细胞分化。     

安氏Ⅲ类错畸形病因的logistic分析

目的研究引起安氏Ⅲ类错牙合畸形的病因。方法对50例安氏Ⅲ类错牙合患者和50例正常牙合人作病因问卷调查,将结果用logistic法分析,提取有效病因。结果共有慢性扁桃体炎、遗传因素、咬上唇3项病因进入方程。结论按贡献大小,长期慢性扁桃体炎、经常咬上唇和遗传因素是导致安氏Ⅲ类错牙合畸形的危险因素。     

C端序列的删除对非出血重组蛇毒纤溶酶的影响

[目的]研究C端序列对非出血重组纤溶酶(rFⅡ)特异性、活性和热敏感性的影响.[方法]通过SOEing PCR方法构建删除C端序列的突变体,突变体和rFⅡ分别在P.pastoris中诱导表达.表达和纯化的蛋白用SDS-PAGE和Westeren blot进行鉴定.之后分析其生化特性.[结果]rFⅡ及突变体通过SDS-PAGE和Westeren blot得到证实.生化分析揭示:①突变体对显色底物N-(p-Tosyl)-Gly-Pro-Lys-pNA的催化效率(Kcat/Km)是rFⅡ的1.9倍;②突变体和rFⅡ对氧化的胰岛素B链拥有共同的优先裂解位点,可是在随后的裂解中开始展现差异;③突变体显示了更高的纤(原)活性;④热处理表明突变体相较r FⅡ对温度的增加更敏感;⑤通过圆二色谱测量,突变体的螺旋比例有所增加.[结论]删除的序列参与rFⅡ的特异性、活性和热敏感性,该序列可作为下一步优化的候选序列.