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.     

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。     

新型穿膜肽介导 p53融合蛋白的表达及其免疫原性分析

目的：原核表达新型细胞穿膜肽RDP介导p53融合蛋白,并检测其免疫原性的强弱．方法：以质粒pET28a‐p53为模板扩增p53基因,并克隆至原核表达载体pET28a‐RDP中,构建重组表达质粒pET28a‐RDP‐p53,转化至大肠杆菌Rosetta ,IPTG诱导表达蛋白并纯化,SDS‐PAGE确定该蛋白准确性．同时,将昆明小鼠分为空白对照组（等容生理盐水）和RDP‐p53融合蛋白高、中、低剂量（4 mg／kg ,2 mg／kg ,1 mg／kg）组,经腹腔注射免疫,每隔2 d给药1次,30 d后眼球取血,用酶联免疫吸附法（ELISA）测定血清中IgG的含量．结果：双酶切及测序结果显示, p53基因已克隆入表达载体中;RDP‐p53融合蛋白在上清和沉淀中均获得表达．ELISA测定结果表明,与对照组比较,低、中剂量组小鼠血清中IgG含量没有明显升高（p＞0.05）,高剂量组显著升高（p＜0.05）．结论：成功表达并纯化RD P‐p53融合蛋白,其免疫原性较弱且与给药剂量相关,为进一步研究RD P‐p53融合蛋白对脑肿瘤的药理作用奠定基础．     

大肠癌中HSP27、p14^ARF和caspase-3蛋白的表达及意义

目的：探讨HSP27、p14^ARF和caspase-3蛋白在大肠癌中的表达及其与大肠癌I临床病理特征的关系。方法：利用sP法检测大肠癌48例和大肠腺瘤10例以及正常大肠黏膜组织15例中HSP27、p14^ARF和caspase-3蛋白的表达。结果：大肠癌组织中HSP27、p14^ARF和caspase-3蛋白阳性表达率分别为79．2％、58．4％和66．7％,大肠腺瘤组织中分别为40．0％、70．0％和100．0％,正常大肠黏膜组织中分别为33．3％、80．0％和93．3％。p14^ARF和caspase-3蛋白的表达在高、中分化癌组的表达率分别为64．1％、74．4％,明显高于低分化组22．2％、33．3％（Pd0．05）。结论：HSP27、p14^ARF和caspase-3与大肠癌的发生有关,可作为大肠癌临床评价肿瘤生物学行为的指标。     

Effects of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus

This study was conducted to evaluate the effects of different concentrations of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus. Diets were formulated to contain 40% crude protein from solvent-extracted menhaden fish meal and 0, 7, 14 or 21% lipid from menhaden fish oil. The basal diet, without supplemental fish oil, contained lipid at 0.4% of dry weight. The diets were fed to groups of 25 juvenile red drum initially averaging 7.3 ± 0.18 g fish^–1 in a recirculating culture system for 8 weeks and weight gain was recorded. After an additional 8 weeks, 16 fish from each treatment were sacrificed and the following measurements were recorded: hepatosomatic index (HSI), intraperitoneal fat (IPF) ratio, and liver -tocopherol, malondialdehyde (MDA) formation, and cytochrome P-4501A activity (measured as 7-ethoxyresorufin O-deethylase (EROD) activity). The activity of alanine and aspartate aminotransferases and concentrations of -tocopherol also were measured in plasma.Weight gain was significantly (p<0.05) affected by dietary lipid concentration, with values ranging from 361% of initial weight for fish fed the basal diet to 527% of initial weight for fish fed the diet containing 7% lipid. The HSI and IPF ratio values also were significantly affected by lipid with the lowest values recorded for fish fed the basal diet and the highest values observed in fish fed the diet containing 21% lipid. Increasing dietary lipid significantly increased oxidative stress as reflected in reduced -tocopherol in liver and plasma and increased MDA formation in the liver, although no overt pathological signs were observed. These findings suggest that lipid concentrations between 7 and 14%, when the diet contains 60 IU vitamin E kg^–1, are likely to limit oxidative stress and result in normal physiological responses of red drum.     

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、...     

Effects of Jiedu-Qingfei mixture on NF-κB and p38 MAPK pathways in lung tissues of rats with Mycoplasma pneumoniae infection

AIM: To observe the therapeutic effect of Jiedu-Qingfei mixture on Mycoplasma pneumoniae (MP)-infected rat lung tissues and to explore its mechanism. METHODS: SD rats (n=40) were randomly divided into 4 groups:blank control group, model group, Jiedu-Qingfei group and positive control group, with 10 rats in each group. The rats in experimental groups were slowly dripped with 1×10⁹ CFU/L MP solution into their nostrils for 4 d. One rat in each group was sacrificed for MP nucleic acid detection at the second day after inoculation, and the other rats were given gavage therapy. The rats in blank control group and model group were intragastrically given the same volume of normal saline, the rats in Jiedu-Qingfei group were given 8 mL/kg Jiedu-Qingfei mixture daily for 4 weeks, and the rats in psoitive control group were given dexmethasone sodium phosphate (0.5 mg·kg^-1·d^-1). After the experiment, the rats were killed. The serum and bronchoalveolar lavage fluid (BALF) were collected for detecting the levels of interleukin-12 (IL-12), IL-13 and TNF-α by ELISA. The right lung tissues were used for pathological observation and HE staining, while the left lung tissues were used to detect the expression of NF-κB p50, I-κBα and p38 mitogen-activated protein kinase (p38 MAPK) at mRNA and protein levels. RESULTS: The results of MP nucleic acid detection showed that all the rats except blank control group were MP nucleic acid positive, indicating that the rat model of MP infection was successfully established. On the 1st day of the treatment, the pathological scores of the lung tissues in model group and Jiedu-Qingfei group were significantly higher than those in blank control group (P<0.05). After treatment, the pathological scores of the lung tissues in mo-del group were significantly higher than those in blank control group and Jiedu-Qingfei group. The levels of IL-12 in the serum and BALF in model group were significantly lower than those in blank control group after MP infection (P<0.05), while those after treatment with Jiedu-Qingfei mixture were significantly higher than those in model group (P<0.05). The levels of IL-13 and TNF-α in the serum and BALF of MP-infected rats were increased significantly, while those after treatment with Jiedu-Qingfei mixture were significantly lower than those in model group (P<0.05). The mRNA expression levels of NF-κB p50 and p38 MAPK in model group were increased significantly (P<0.01). After treatment, the mRNA expression levels of NF-κB p50 and p38 MAPK were decreased significantly compared with model group (P<0.01). The mRNA expression level of I-κBα in model group was significantly lower than that in control group. After treatment, the mRNA expression of I-κBα in Jiedu-Qingfei group was significantly higher than that in model group (P<0.05). The protein levels of NF-κB p50 and p38 MAPK in the lung tissues of model group were significantly higher than those of blank control group. After treatment, the protein expression of NF-κB p50 and p38 MAPK was decreased significantly. The protein level of I-κBα in model group was significantly lower than that in blank control group, and after treatment with Jiedu-Qingfei mixture, the protein expression level of I-κBα was increased significantly (P<0.05). CONCLUSION: Jiedu-Qingfei mixture may attenuate lung tissue inflammation caused by MP through NF-κB and p38 MAPK pathways.     

猪miR-199a-5p对脂质生成的影响及其靶基因的预测和分析

本研究旨在分析miR-199a-5p对猪肌内脂肪细胞脂质生成的影响及作用机制。采集淮南猪不同育肥时期(育肥前期、中期和后期)的背最长肌和皮下脂肪组织,通过实时荧光定量PCR分析其表达变化趋势;合成miR-199a-5p的mimics,转染猪原代肌内脂肪细胞,诱导分化后通过油红O染色观察过表达miR-199a-5p对脂质生成的影响;结合之前筛选到的脂肪型和瘦肉型猪差异表达mRNAs、lncRNAs和circRNAs,使用miRanda软件筛选miR-199a-5p的靶分子,并用GO和KEGG富集分析法对这些mRNAs、lncRNAs的共表达基因和circRNAs的来源基因进行功能分析。结果显示,随着育肥进程,miR-199a-5p在背最长肌中表达量持续上调,而在皮下脂肪组织中表达量先下调后上调;过表达miR-199a-5p可抑制肌内脂肪细胞的脂质生成。共筛选到9个mRNAs、5个lncRNAs和1258个circRNAs包含miR-199a-5p结合位点,GO分析主要富集于内质网和肌动蛋白结合,KEGG分析发现主要富集于糖、脂质和蛋白质代谢。提示miR-199a-5p可能通过与lncRNAs和circRNAs互作间接调控靶基因,参与调控肌肉发育、脂质生成和代谢,可作为影响肉质性状的候选miRNA。     

Cyclooxygenase (COX) inhibitors and the intestine

Nonsteroidal anti-inflammatory drugs (NSAIDs) have long been used for the treatment of pain and inflammation because of their inhibitory effects on cyclooxygenase (COX). For almost as long as NSAIDs have been in use, multiple adverse effects have been noted. Assessment of many of these adverse effects have been complicated because of the discovery of multiple splice variants of the cox gene, and a greater array of COX inhibitors, especially the COX-2 selective inhibitors have become available. Some of these adverse effects cannot be readily explained by the effect of these drugs on COX. This has sparked a new field of investigation into the COX-independent effects of the COX inhibitors. The major noncyclooxygenase targets of the COX inhibitors of particular relevance to inflammation and the gastrointestinal tract are phosphatidylinositol 3'-kinase Akt signaling, uncoupling of oxidative phosphorylation, PPARgamma, nuclear factor KB, mitogen activated protein kinases, and heat shock proteins.