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.     

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.     

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。     

优质杂交晚籼稻新组合乐优97的选育及栽培制种技术

乐优97是一种典型的晚籼型三系杂交水稻良种,于2022年通过中国国家农作物品种审定委员会审定,系由合肥丰乐种业股份有限公司所属的水稻研究院通过自己创制的三系不育系409A与恢复系R977 杂交选育而成的高产、优质、抗倒杂交晚籼型新型组合。文中详细尽介绍了乐优97的选育步骤,重点介绍了乐优97的特征特性,并归纳出了乐优97优质高产栽培技术及其制种技术关键要点,希望为该产品的以后大规模推广种植打下基础。     

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、...     

普通小麦-柔软滨麦草易位系M97抗条锈基因的遗传分析和分子作图

为了明确M97抗条锈性遗传规律,在苗期用7个小麦条锈菌系对M97与感病品种铭贤169的杂交后代F1、F2、F3和BC1代进行抗条锈性遗传分析,并对M97抗Sun11-4的抗条锈基因进行SSR分子标记。M97对Sun11-4和Sun11-11的抗病性均由1对显性基因控制,对CY29、CY30、CY33的抗病性由1显1隐2对基因共同控制,对CY31的抗病性由2对显性基因独立或重叠作用控制。以接种Sun11-4的F2代分离群体构建作图群体,筛选到Xwmc222、Xwmc147、Xbarc229和Xwmc339等4个与抗病基因连锁的SSR标记,其遗传距离分别为3.4、4.8、7.6和12.1 cM。将该抗病基因定位于小麦1DS染色体,且该基因不同于已知的抗条锈基因,暂命名为YrM97。用YrM97两侧遗传距离最近的2个标记Xwmc222和Xwmc147对42个黄淮麦区主栽小麦品种进行分子检测,仅有9.5%的品种具有与YrM97相同的标记位点。     

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.     

龙岩市地方优良种禽原种场禽白血病p27抗原检测与分析

用酶联免疫吸附试验法对长汀河田鸡原种场、武平象洞鸡原种场、龙岩山麻鸭原种场共686份蛋清样本及801份泄殖腔棉拭子进行禽白血病p27抗原检测。结果为：龙岩山麻鸭的蛋清样本和泄殖腔棉拭子均未检出ALV核酸;鸡蛋清样本阳性率为12.8%（80/626）,明显高于泄殖腔棉拭子阳性率8.9%(66/741);鸡原种场180日龄蛋清样本和泄殖腔棉拭子阳性率均最高,平均阳性率分别为15.0%和11.3%。表明：龙岩市境内地方优良鸡原种场中有禽白血病感染现象。