Relationship of p21WAF1 gene polymorphisms with protein expression in breast carcinomas

AIM: To investigate the relationship between p21^WAF1gene polymorphisms and protein expression in breast carcinoma. METHODS: Polymerase chain reaction single-strand conformation polymorphisms technique (PCR-SSCP) and immunohistochemical assay of S-P immunostaining technique were used to study polymorphisms of p21^WAF1 and protein expression respectively on the specimen of paraffin-embedded tissues in 100 cases of breast carcinomas and 40 benign breast diseases as control. RESULTS: Two p21^WAF1 gene polymorphisms were found in 18% (18/100) of breast carcinomas and 5% (2/40) of control samples. The difference between the two groups was statistically significant (χ²=3.94, P＜0.05). The positive immunohistochemical reaction of p21^WAF1 protein were found in 50% (50/100) of breast carcinomas and 12.5% (5/40) of control samples. The difference between the two groups was statistically significant (χ²=16.84, P＜0.01). The positive immunohistochemical reaction of p21^WAF1 protein were found in 100% (18/18) of breast carcinomas with p21^WAF1 gene polymorphisms and 39% (32/82) of no p21^WAF1 gene polymorphisms. The difference between two groups was statistically significant (χ²=21.95, P＜0.01). The p21^WAF1 gene polymorphisms were correlated with the protein expression in breast carcinomas (r=0.576, P＜0.01). CONCLUSION: p21^WAF1 gene polymorphisms may create the different copies of mRNA and may make relevant protein molecules.     

法氏囊活性肽BP7调节鸡未成熟B细胞的基因表达谱及功能分析

试验旨在探索法氏囊活性肽BP7调节鸡未成熟B细胞的分子基础.利用BP7刺激禽前B淋巴细胞DT40细胞,采用荧光定量PCR(qPCR)检测IgM的mRNA水平,并采用基因芯片分析基因表达谱及其生物学功能.结果 显示,BP7刺激的DT40细胞产生IgM的mRNA水平明显升高.基因芯片分析发现,BP7处理的DT40细胞中共有...     

miR-186-5p与α-MSH互作对绵羊黑素细胞黑色素生成的影响

黑色素合成受多种基因和miRNAs的调控,为了研究miR-186-5p与α-MSH协同作用对绵羊黑素细胞黑色素生成的影响.本研究在绵羊黑素细胞中转染miR-186-5p表达载体,同时添加α-MSH,通过实时荧光PCR(quantitative real time polymerase chain reaction,qR...     

卵黄和低密度脂蛋白对冷冻保存绵羊精子胆固醇、磷脂含量的影响

为了探索绵羊冷冻精液受精率低的原因,在稀释液中用低密度脂蛋白代替卵黄冷冻绵羊精液,研究LDL对冷冻中精子脂类含量的影响。结果表明,以卵黄稀释液冷冻精液,精子胆固醇含量从鲜精的234．54nmol／10^9降低到163．76nmol／10^9精子,胆固醇与磷脂的比值（c／p）从0．54下降到0．36;以9g／100mL LDL代替卵黄,冷冻后胆固醇含量为243．73nmol／10^9精子,c／p值保持在0．50。研究得出在卵黄稀释液中冷冻绵羊精液,精子胆固醇流失,c／p值下降;以LDL代替卵黄,可有效保护精子胆固醇,避免流失,维持了精子c／p值。     

MicroRNA-142-3p modulates atherosclerosis-associated endothelial cell apoptosis via targeting Rictor

AIM To investigate the role of microRNA-142-3p （miR-142-3p） in endothelial cell apoptosis during atherosclerosis （AS） and the underlying mechanism. METHODS Human aortic endothelial cells （HAECs） were treated with oxidized low-density lipoprotein （ox-LDL）. The expression level of miR-142-3p was detected by RT-qPCR. Apoptosis was determined via flow cytometry （FCM） and caspase-3 activity assay. Prediction of the binding site between miR-142-3p and 3’-UTR of Rictor mRNA was performed by bioinformatics analysis and confirmed by dual-luciferase reporter assay. RESULTS The expression of miR-142-3p was substantially up-regulated during the ox-LDL-elicited apoptosis in HAECs （P<0.05,P<0.01）. Forced expression of miR-142-3p exacerbated apoptosis in HAECs whereas inhibition of miR-142-3p partly alleviated apoptotic cell death mediated by ox-LDL. Further analysis identified Rictor as a direct target gene of miR-142-3p, and Rictor knock-down abolished the anti-apoptotic effect of miR-142-3p inhibitor. Moreover, the Akt/endothelial nitric oxide synthase （eNOS） signaling pathway was found to mediate the beneficial effect of miR-142-3p inhibitor on endothelial cells apoptosis. CONCLUSION Down-regulation of miR-142-3p inhibits endothelial cell apoptosis and atherosclerotic development by up-regulating the expression of Rictor and activating the Akt/eNOS signaling pathway.     

花椰菜新品种夏雪40的选育

夏雪４０是以９１Ａ－Ｃ－２－Ｄ１３－９－１５为母本,９１Ｂ－Ｃ－１－２－５－８为父本配制的一代杂种,极早熟,成熟期４５天左右,株型直立紧凑,花球洁白,较紧实,平均单球重０．４５ｋｇ,耐高温高湿,每６６７ｍ＾２产量１４８０￣１６８０ｋｇ,比对照白峰早熟７天左右。     

热研4号王草干燥特性的试验研究

在将热研4号王草调制成青贮饲料或干草的加工中,干燥是一个关键性环节,也是发展热研4号王草产业的瓶颈问题。为此,采用快速薄层干燥的试验方法,研究热研4号王草的含水量与干燥时间、干燥介质温度两者之间的关系,并分析影响其营养成分的因素。结果表明:热研4号王草的干燥最佳介质温度为150°～180℃,鲜草刈割后必须在24h内进行快速干燥。     

静原鸡let-7a-5p的靶基因预测及组织表达分析

【目的】了解let-7a-5p的生物学信息,初步探索其在静原鸡肌肉组织中的功能作用。【方法】利用生物信息学方法对转录组测序获得的静原鸡let-7a-5p的成熟序列进行保守性分析,使用miRDB、TargetScan和miRanda在线软件预测let-7a-5p的靶基因,取其交集进行GO功能和KEGG通路富集分析,并利用实时荧光定量PCR检测let-7a-5p在静原鸡公鸡和母鸡各组织中的表达水平。【结果】let-7a-5p的成熟序列在各物种间高度保守,共预测得到38个基因集合。在生物过程、细胞组分和分子功能中靶基因富集较多的GO条目有细胞过程、生物调节、细胞、结合和催化活性;靶基因显著富集于聚酮糖单元生物合成、AGE-RAGE信号通路在糖尿病并发症中的作用、泛素介导的蛋白水解、TGF-β信号通路和加压素调节的水重吸收信号通路,且富集最多的是代谢途径、AGE-RAGE信号通路在糖尿病并发症中的作用、泛素介导的蛋白水解和MAPK信号通路。let-7a-5p-靶基因互作主要通过作用于靶mRNA的3′-UTR抑制表达或使其降解。组织表达发现,let-7a-5p在静原鸡公、母鸡的心脏、肝脏、脾脏、...     

Increased expression of CD40 ligand by peripheral blood mononuclear cells in patients with systemic lupus erythematosus

AIM:To investigate expression and function of CD40 ligand by peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE).METHODS:Expression of CD40 ligand by PBMCs in patients with SLE and control were examined by flow cytometric analysis before and after stimulated by phytohemagglutinin(PHA)and depressed by Dexamethasone(Dex). The correlation between expression of CD40 ligand and SLE activity index(SLEDAI) was analysed in patients with SLE.RESULTS:The expression of CD40 ligand by PBMCs in patients with active SLE was higher than that in patients with inactive SLE and control. Though the expression of CD40 ligang by PBMCs could be stimulated by PHA in three groups, it was the highest in patients with active SLE. Dex depressed the expression of CD40 ligand by PBMCs significantly in patients with SLE, but not in control. There was high positive correlation between expression of CD40 ligand and SLEDAI in patients with active and inactive SLE.CONCLUSION:Increased expression of CD40L by PBMCs in patients with SLE may play an important role in pathogenesis of SLE.     

牛朊蛋白和酵母朊毒体结构域融合蛋白的原核表达及其单克隆抗体的制备

利用DNA重组技术将酵母Ure2p朊蛋白结构域(UPD)基因插入牛朊蛋白(BPrP)表达质粒pET-PrP中,构建原核表达载体pET-PrP-UPD.阳性质粒转化宿主菌BL21 (DE3),在IPTG诱导下获得高效表达.Westernblot检测表明表达的重组蛋白与单克隆抗体6H4呈现特异性反应,且纯化的PrP-UPD融合蛋白在体外具有聚集成淀粉样纤维、抵抗蛋白酶K消化等朊毒体的结构特点.利用纯化的PrP-UPD免疫BALB/c小鼠,取其脾细胞与SP2/0细胞融合,经克隆和筛选,获得3株稳定分泌抗重组PrP单抗的杂交瘤细胞,分别命名为1G3、3A4、4D1,其中1G3分泌的单抗能识别细胞型牛朊蛋白,其Ig亚类为IgG2b,腹水ELISA效价为1×105,Western blot表明该单抗具有较强的特异性.