Effect of DRB on the Biological Characteristics of Human Laryngeal Carcinoma Hep-2 Cell Line

In order to study the effect of 5, 6-Dichloro-1-β-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68±0.19)%, (1.95±0.12)%, (8.51±0.26)%, (11.26±0.17)% and (14.99±0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concen-tration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.     

天蓝淡红链霉菌SIPI-1482中dnrX基因的敲除及多柔比星产生菌的构建

柔红霉素是合成重要抗肿瘤抗生素多柔比星的前体,由微生物发酵产生。通过PCR从国内柔红霉素生产菌种天蓝淡红链霉菌SIPI-1482基因组DNA中扩增出约1080bp的dnrX部分序列,将PCR扩增片段中985bp的片段亚克隆至大肠埃希菌质粒pYG813构建了同源重组突变质粒pYG963,转化SIPI-1482原生质体并成功地敲除了dnrX基因,得到一株稳定的突变株。该突变株的发酵试验表明dnrX基因所编码的酶的功能已经被有效地阻断,对酸敏感的副产物减少,柔红霉素的产量增加了近3倍,将原野生菌改造成为多柔比星产生菌,发酵效价与原野生菌株柔红霉素的发酵效价相当,为直接发酵法生产多柔比星打下了基础。     

Involvement of p38 mitogen-activated protein kinase in the regulation of platelet-derived growth factor induced cell migration

The aim of this study was to investigate the role of p38 mitogen-activated protein kinase (MAPK) in cell migration induced by platelet-derived growth factor (PDGF). Western blot was performed to detect the phosphorylation of p38 in NIH3T3 cells treated with PDGF. A Transwell cell migration system was used to determine the effects of PDGF treatment on the migration of NIH3T3 cells and the influence of p38 deficiency on this process in a p38 gene knockout (p38^−/−) mouse embryonic fibroblast cell line. On the stimulation of PDGF, the migration of NIH3T3 cells was significantly increased (P < 0.001) compared to the control and p38 MAP kinase was simultaneously phosphorylated. Furthermore, the PDGF-induced cell migration was significantly blocked in p38 gene knockout (p38^−/−) mouse embryonic fibroblasts (MEFs) (P < 0.001) as compared with the wild type cells (p38^+/+). p38 MAPK plays an important role in the regulation of cell migration induced by PDGF. Translated from Medical Journal of Chinese People’s Liberation Army, 2007, 32(3): 205–207 [译自: 解放军医学杂志]     

小鼠肝枯否细胞的分离、培养与鉴定

目的研究BALB／c鼠肝枯否细胞分离、培养和鉴定。方法选用健康雌性8—10w龄鼠，麻醉无菌取肝，D—Hank’s液注射冲洗去血并剪去部分结缔组织，剪碎肝组织，加入链霉蛋白酶E和Ⅰ型胶原酶，水浴振荡消化，再加入DNase Ⅰ，共同消化。不锈钢筛网滤过，用0．005％DNase Ⅰ的1640液清洗细胞液，加Tris—NH4CL溶解红细胞。50％-70％Percoll梯度离心，用含10％FCS的RPMI-1640贴壁培养4—6h洗去非贴壁细胞。通过吞噬墨水实验鉴定枯否细胞。结果枯否细胞的数量为（1．07±0．87）×10^7／g，贴壁率40．3％，用0．4％台盼蓝染色鉴定，细胞存活率在95％以上，吞噬实验发现90％的贴壁细胞具有吞噬功能，即KC的纯度可达90％。贴壁的枯否细胞的形态多样，已经完全伸出了伪足并且伸展贴壁，呈多边形、多角形或者梭形等不规则形状，并且胞浆内可见吞噬的微粒状物质。结论酶消化水浴振荡法结合梯度离心和贴壁培养是分离BALB／c鼠枯否细胞的可靠方法，为进一步实验打下了基础。     

大鼠卵巢促性腺激素释放激素(GnRH)受体及其特性

采用乳过氧化物酶-葡萄糖氧化酶法放射性碘标记国产GnRH激动剂D-色~6-GnRH,用阴离子交换柱层析分离纯化,自身置换曲线计算出标记物比活性为700～1000μCi/μg。~(125)Ⅰ-D-色~6-GnRH与大鼠卵巢细胞膜有特异性结合,结合不受P物质、胰高糖素、生长素释放抑制素、促甲状腺素释放激素、血管紧张素Ⅱ等的竞争抑制。~(125)ⅠD-色~6-GnRH与肾皮质、骨骼肌、心肌等没有特异性结合,与大鼠胎盘有部分结合。大鼠卵巢GnRH受体与垂体GnRH受体的亲和常数基本相同,Ka2.43×10~9M~(-1);结合位点为垂体的20%左右,46.06fm/mg蛋白。本实验表明大鼠卵巢存在着特异性的GnRH受体,有助于阐明GnRH及其类似物的垂体外作用机理。     

三维超声心动图左室容积值与左室造影参数的相关性分析

应用国内三维超声心动图(3DE)计算机软件系统,测定20例正常人及40例心血管病患者的左室容积值,观察各组心血管病左室长轴,短轴内壁周长及周长缩短率的变化,并就10例患者进行了3DE与左室造影心功能参数的相关性分析。结果表明,正常组与各心血管病组左室容积值差异显著;差异的内在规律集中在心室短轴缩短率的变化上;3DE测定的左室容积值与左室造影所测参数比较,相关性密切。     

不育或流产患者抗精子及抗附睾的人单克隆抗体制备

我们用ＥＢ病毒转化法从免疫性不育或流产患者的淋巴细胞中制备了一批抗精子和抗附睾的人单克隆抗体。抗精子抗体与全精子、精子头、精子尾、赤道部或核后部反应；抗附睾抗体与附睾基细胞或纤毛区反应。这些来源于不育患者淋巴细胞的抗体具有抗生育效应的可能性较大，对人体具有副作用的可能性相对较小。其中抗附睾基细胞抗体的作用区段与精子在附睾内成熟的区段相一致，这种抗体对基细胞的功能及其在生殖过程中的作用均有特殊的研究价值。     

Experimental study of the therapeutic effect of You-Gui-Wan on hypogonadism in rats

Rats raised on a diet containing 0.5% adenine showed atrophy of the testes and retrogressive degeneration of the convoluted seminiferous tubules. The development of spermatids was suppressed, and the cell count decreased. In these rats, the activity of 3β-hydroxysteroid dehydrogenase was markedly decreased, and the blood levels of testosterone and oestradiol were also very low. However, in rats that were also given You-Gui-Wan, retrogressive degeneration of the testes was milder, and the blood levels of testosterone and oestradiol were higher, indicating the beneficial effects of You-Gui-Wan on the histomorphology of the testes.     

Apoptosis and related gene expression in lacrimal gland of cases with Sjögren's syndrome

OBJECTIVE: To assess the roles of apoptosis and expression of the related genes in lacrimal glandular destruction of patients with Sj?gren's syndrome (SS). METHODS: Small pieces of palpebral lobes of lacrimal glands were obtained from 12 patients (10 with primary Sj?gren's syndrome, 2 with secondary Sj?gren's syndrome) and 7 normal controls. They had been investigated by the in situ end labeling and immunohistochemical staining to detect the apoptotic cells and the expression of Fas, FasL, bcl-2 and Bax. RESULTS: The number of epithelial apoptotic cells in lacrimal glands of patients with SS was significantly higher than that in normal glands, and SS epithelial cells showed increased expression of Fas, FasL and Bax. There was significant positive correlation between the number of the apoptotic cells and that of the cells expressing Fas, FasL and Bax, respectively, and there was significant negative correlation between the number of the apoptotic cells and that of the cells expressing bcl-2. CONCLUSIONS: The apoptosis of the epithelial cells in the lacrimal gland may be one of the mechanisms leading to the glandular destruction found in SS. In SS lacrimal glands, the expression of Fas, FasL and Bax may promote apoptosis, and the expression of bcl-2 may inhibit apoptosis.