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.     

胰岛素样生长因子-Ⅰ和胰岛素样生长因子结合蛋白-1与妊高征的关系

目的：探讨胰岛素样生长因子 I(IGF-Ⅰ)和胰岛素样生长因子结合蛋白 1(IGFBP—1)与妊高征(PIH)的关系。方法：采用免疫放射法测定50例妊高征孕妇(PIH组)和108例正常孕妇(对照组)的血清 IGF-Ⅰ、IGFBP-1水平，并对其结果进行相关性分析。结果：①妊高征孕妇血清 IGF-Ⅰ水平低于正常孕妇(P<0.05)，且重度妊高征孕妇血清 IGF-Ⅰ水平低于中度和轻度孕妇(P＜0.05，P＜0.01)。②妊高征孕妇血清 IGFBP-1水平高于正常孕妇(P＜0.05)，且重度妊高征孕妇血清 IGFBP-1水平高于中度和轻度孕妇(P＜0.05，P＜0.01)。③两组孕妇血清 IGF-Ⅰ水平与血清 IGFBP-1水平呈负相关(r=-0.386，P＜0.05)。④妊高征孕妇血清 IGF-Ⅰ水平与舒张压呈负相关(r=-0.386，P＜0.05)，血清 IGFBP-1水平与舒张压呈正相关(r=0.632，P＜0.01)。结论：妊高征孕妇血清 IGF-Ⅰ、IGFBP-1水平的高低可反映妊高征的严重程度。     

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.     

胰岛素样生长因子-1基因转染脂肪间充质干细胞向软骨细胞分化的研究

目的 观察胰岛素样生长因子(IGF)-1基因转染的脂肪间充质干细胞(ADSCs)向软骨细胞分化的效果.方法 原代培养兔ADSCs,免疫荧光法检测细胞表面抗原CD44、CIM9;脂质体介导人IGF-1基因转染兔ADSCs联合低浓度血清培养基向软骨细胞分化诱导,RT-PCR及Western blot方法检测IGF-1的表达,MMT法绘制细胞增殖曲线、甲苯胺蓝染色软骨结节、免疫组织化学检测Ⅱ型胶原的表达.结果 脂肪间充质干细胞CD44、CD109表达阳性,基因转染后细胞IGF-1表达阳性,细胞增殖速度增快,出现软骨结节,Ⅱ型胶原表达增高.结论 从脂肪组织中能够分离出增殖旺盛的ADSCs,IGF-1在ADSCs内获得稳定表达,细胞增殖能力增强,促进其向软骨细胞分化.     

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.     

转化生长因子β1对人牙髓细胞微丝骨架的重组作用

目的:了解转化生长因子β1(TGF-β1)对人牙髓细胞微丝和微管骨架的作用。方法:I型胶原酶消化组织块法体外培养人牙髓细胞,以20ng/ml的TGF-β1处理细胞,分别在第30min、1、6和24h收集细胞爬片,BODYPY-Phalloidin对微丝作直接荧光染色、Rhodamine RedTM对微管蛋白α(tubulin-α)作间接免疫荧光染色,采用激光扫描共聚焦显微镜观察TGF-β1作用于牙髓细胞后不同时间点微丝和微管的变化情况。结果:TGF-β1作用于人牙髓细胞后,微丝出现解聚重组现象,在30min时间点,肌动蛋白(actin)在细胞膜下聚合成纤维形肌动蛋白F-actin,同时胞质内的F-actin解聚,6h解聚最明显,24h后可见胞质内微丝重组。观察的各时间点,微管结构未见明显解聚重组现象。结论:TGF-β1能够使牙髓细胞微丝骨架重组。     

大肠癌患者血清MMP-9与VEGF水平的变化及意义

目的本文检测血管内皮生长因子(VEGF)和基质金属蛋白酶(MMP-9)在大肠癌患者血清中的水平,探讨其与大肠癌恶性程度、浸润、转移的关系。方法用ELISA法对48例大肠癌患者及40例正常对照者血清VEGF和MMP-9进行检测。结果大肠癌患者血清VEGF、MMP-9的水平分别为(665.8±232.8)ng/L;(532.7±208.9)ug/L。显著高于对照组(391.6±135.2)ng/L;(317.8±132.2)ug/L(P<0.01)。并与组织分化程度、Duke’s分期有关。低分化者及C、D期患者较中、高度分化及A、B期高(P<0.01)。结论大肠癌患者血清VEGF和MMP-9水平增高,与肿瘤的组织分化程度、转移、分期有关,提示VEGF、MMP-9对大肠癌恶性程度的判断有一定的价值。