重组人白细胞介素10对角朊细胞增殖及产生细胞因子的影响

目的：研究重组人白细胞介素10(rhIL-10)对无血清培养的角朊细胞增殖和产生细胞因子的影响，探讨其治疗银屑病的作用机制。方法：用四甲基偶氮唑蓝比色法(MTT)测定其对细胞增殖的影响；小鼠胸腺细胞增殖法检测白细胞介素1(IL-1)；ELISA法检测白细胞介素6(IL-6)、白细胞介素8(IL-8)。结果：重组人白细胞介素10抑制角朊细胞增殖与IL-1、IL-6及IL-8的分泌，并呈剂量依赖关系。结论：重组人白细胞介素10抑制角朊细胞与细胞因子分泌，可能是治疗银屑病的作用机理之一．     

M-CSF胞质内稳定表达细胞系的构建和鉴定

目的构建真核细胞pCMV/cyto/myc-M-CSF载体,建立M-CSF胞质内稳定表达细胞系,为进一步研究M-CSF的胞内作用奠定基础。方法构建靶向定位载体pCMV/cyto/myc-M-CSF,PCR及双酶切鉴定后转染HeLa细胞,G418筛选阳性克隆,RT-PCR和免疫细胞化学鉴定M-CSF的表达及M-CSF的蛋白定位。结果重组质粒pCMV/cyto/myc-M-CSF用M-CSF特异性的引物进行PCR扩增,结果显示插入片段约为1 400 bp,双酶切后分别得到约5.0 kb和1 400 bp的两条带,M-CSF分子大小与预期一致。RT-PCR、免疫细胞化学结果表明转染M-CSF的HeLa细胞高表达M-CSF（P〈0.05）,并且转染M-CSF的HeLa细胞表达的M-CSF蛋白定位于细胞质。结论成功构建了稳定高表达胞质M-CSF的HeLa细胞系。     

浸润性乳腺癌组织上皮钙依赖粘附素相关分子的表达

目的探讨乳腺浸润性小叶癌(ILC)和浸润性导管癌(IDC)中上皮钙依赖粘附素相关分子α-cat、β-cat、γ-Cat的表达及其意义.方法采用免疫组织化学SP法检测38例ILC(其中有淋巴结转移17例)和64例IDC(有淋巴结转移14例)组织中α-Cat、β-Cat、γ-Cat的表达,按阳性癌细胞占肿瘤细胞的比例进行半定量分析,结果38例ILC中α-Cat、β-Cat、γ-Cat表达缺失和低表达者分别为32例(84.21%),22例(57.89%)和33例(86.84%),而64例IDC癌组织中3者的表达缺失和低表达者分别为50例(90.62%),5例(7.81%)和52例(81.25%),α-Cat与β-Cat在ILC和IDC中的表达具有明显的正相关性(P＜0.05);β-Cat的表达缺失和低表达者在乳腺ILC和IDC与有无淋巴结转移呈负相关性(P＜0.05),而α-Cat、γ-Cat在乳腺ILC和IDC的表达与有无淋巴结转移无关(P＞0.05).结论α-Cat、β-Cat、γ-Cat在乳腺ILC和IDC中表达均明显减少和缺失,提示上皮钙依赖粘附素相关分子在乳腺浸润性癌发生发展中丧失了其正常的细胞粘附功能.β-Catenin的表达缺失和低表达可能与乳腺癌的淋巴结转移有关.     

神经干细胞分化研究进展

神经干细胞具有比预期广的分化能力。要充分发挥神经干细胞的潜能,必须揭示其向特异性细胞系分化的机制。近来研究证实,细胞因子及内环境信号均影响神经干细胞的分化方向。对神经干细胞分化机制认识的加深,将大大加速神经干细胞的临床应用。     

Direct intrauterine sampling with Uterobrush: cell preparation by the "flicked" method

The aim of this study was to evaluate the efficacy of endometrial cytology using the Uterobrush in the collection of samples for the diagnosis of endometrial lesions. The preparation technique for endometrial brushing specimens was also demonstrated. In their earlier study, the authors described the cyto-architectural criteria that appear to be more useful for the cytological assessment of endometrial lesions. Therefore, for the application of the diagnostic criteria, endometrial cytological sampling will become more important. With regard to the cell sample collection, the authors used the Uterobrush because the insertion into the uterine cavity is easy and painless. The authors compared the Uterobrush with the Endocyte, since they thought that cell clumps using the former device tended to be smaller. The purpose of the current study was to improve and evaluate cell preparation methods using the Uterobrush. The authors investigated three methods [i.e., conventional and improved techniques ("flicked" method) with the Uterobrush and the Endocyte] of endometrial cell collection and preparation. Using conventional methods, a brush was rolled along a glass slide and the collected material spread and smeared. However, using the "flicked" method, a brush is strongly flicked with forceps, so that the cells are transferred to the slide and its position is moved along the slide little by little and smeared. The frequency by size of cell clumps with tube or sheet-shaped pattern was examined in the preparations. Cell block specimens with the Uterobrush were also investigated. Endometrial cytology from a total of 90 women was evaluated. Most were outpatients and all were older than 20 yr (ranging from 20 to 54, average 42.7 yr). Of these, 30 cases from each group were examined by three methods.Uterobrush samples prepared by the "flicked" method have a greater quantity of cell clumps than those using the Endocyte sampler, while the frequency-by-size of cell clumps was by degree the same as the Endocyte. The cell clumps obtained in the Uterobrush "flicked" method preparation was considered equivalent or superior as an aid to making a diagnosis of endometrial lesions and it became obvious that the same criteria were applicable to both of instruments. Our cytological examination may be a potent aid to making a diagnosis of endometrial lesions and these findings will be helpful in the standardization of criteria in direct intrauterine cell samples.     

The leader sequence triggers and enhances several functions of clusterin and is instrumental in the progression of human prostate cancer in vivo and in vitro

OBJECTIVE: To investigate the role of the leader sequence (which during clusterin biosynthesis facilitates its proper post-translational processing and secretion) in the functional activities of clusterin, a ubiquitous secretory glycoprotein with many biological functions, reported to be pro-apoptotic and anti-apoptotic in target cells, but for which the dual mechanism remains unclear. MATERIALS AND METHODS: We designed an expression vector starting from the second in-frame ATG on the full-length human clusterin cDNA that was capable of driving the expression of both the full-length and the truncated isoforms of clusterin. We established stable expression clones of the androgen-dependent prostate cancer line LNCaP expressing clusterin with and without the leader sequence. This induced expression provided an opportunity to evaluate both the in vivo and in vitro actions of clusterin expression. RESULTS: The LNCaP cells expressing clusterin with the leader sequence resisted apoptosis induced by tumour necrosis factor (TNF)-alpha, but clones with no leader sequence were highly susceptible to TNF-alpha-induced apoptosis. Furthermore, in the absence of the leader sequence, the expressed clusterin had a molecular weight consistent with that of the predicted holoprotein (40 kDa), suggesting a compromised post-translational processing with diffuse distribution throughout the cytoplasm. However, cells transfected with the full-length vector expressed clusterin of 60 and 35 kDa variants, and located exclusively in the Golgi apparatus. In vivo, only the overexpression of the full-length clusterin is anti-apoptotic and stimulates the proliferation of tumour. CONCLUSION: The leader sequence is important in determining the functions of clusterin, which include anti-apoptotic and anti-necrotic properties. The lack of the leader sequence allowed the incompletely processed clusterin to induce apoptosis in target cells; without the leader sequence, clusterin functions differently. Thus, the leader sequence is a trigger for many functions of clusterin in the progression of human prostate cancer cells.     

塞隆骨提取物对牛Ⅱ型胶原诱导的小鼠关节炎的治疗作用及机制研究

目的了解塞隆骨水提物(SLG-B)对胶原诱导的小鼠关节炎的治疗作用及治疗机制。方法利用牛Ⅱ型胶原诱导DBA/1小鼠类风湿性关节炎模型即CIA。SLG-B口服给药观察小鼠关节炎指数的变化情况,体外培养关节炎小鼠脾细胞及巨噬细胞,ELISA法测定IL-12、IL-2、IFN-γ、TNF-α几种细胞因子。RT-PCR测定SLG-B对关节炎小鼠巨噬细胞IL-1β、IL-6、iNOS mRNA表达的影响。结果SLG-B能明显的抑制牛Ⅱ型胶原诱导关节炎的发生,能够减轻关节炎的各种症状。SLG-B在加抗原和不加抗原的情况下都能够明显的抑制关节炎小鼠脾细胞的增殖同时发现SLG-B能够抑制IL-2、IFN-γ、L-12p40、TNF-α细胞因子的产生还能够抑制小鼠腹腔巨噬细胞IL-1、IL-6及iNOS mRNA的表达。这可能是SLG-B在小鼠关节炎中表现治疗效果的分子基础。结论SLG-B对小鼠关节炎有很好的治疗效果,其作用机制主要是通过抑制巨噬细胞及脾细胞产生炎性细胞因子和抑制炎性细胞因子mRNA的表达来达到治疗类风湿性关节炎的作用。     

尿路上皮癌诊断中 DNA倍体分析和尿脱落细胞学检查的临床价值

目的：探讨DNA倍体分析和尿脱落细胞学检查对尿路上皮癌诊断的临床价值,寻找早期诊断尿路上皮癌的理想方法。方法随机收集121例尿路上皮癌患者的尿液标本作为实验组,另取95例良性血尿非肿瘤患者的尿液标本作为对照组,分别进行尿脱落细胞学检查和DNA倍体分析。结果尿脱落细胞学检查的敏感性高于DNA倍体分析,但特异性低于DNA倍体分析。尿脱落细胞学检查敏感性为85.10%,特异性为76.80%;DNA倍体分析敏感性为81.80%,特异性为81.10%。121例中,膀胱癌103例,上尿路上皮癌18例。对于不同分型的尿路上皮癌,两种方法在高级别肿瘤中均有较好的敏感性,尤其是DNA倍体分析在浸润性膀胱癌和上尿路上皮癌的诊断中具有极好的敏感性,均>94%。结论 DNA倍体分析结合尿脱落细胞学检查,可大大提高尿路上皮癌检测的特异性,以及增加对浸润性膀胱癌及上尿路上皮癌诊断的灵敏度,是对尿路上皮癌检测方法的有利补充。     

枸橼酸碳酸氢盐血液透析液对外周细胞因子和NOS的影响

目的:研究枸橼酸碳酸氢盐血液透析液对外周细胞因子和一氧化氮合成酶的影响。方法:对30例维持性血液透析的患者采取自身前后对照研究,前4周行醋酸碳酸氢盐透析液透析(对照组),后4周行枸橼酸碳酸氢盐透析液透析(实验组)。观察透析前后单核细胞数量的变化;透析前、后及第4周透析后,测定外周血IL-1β、TNF-α及TNOS和iNOS的吸光度,分别计算TNOS和iNOS的活力。结果:透析后实验组单核细胞数明显下降;透析前两组TNF-α、IL-1β、TNOS和iNOS水平没有差异,透析后及第4周透析后,对照组增高均较实验组明显。结论:枸橼酸碳酸氢盐血液透析液能减少透析过程中单核细胞和内皮细胞功能的激活。