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.     

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.     

Mechanism of Chinese processing in patients with Broca aphasia

AIM: To explore the mechanism of Chinese processing in patients with Broca aphasia so as to offering accordance for speech rehabilitation. METHODS: The test of orthographic, semantic information and phonological were conducted. The scores in each group with Chi-square test in patients were compared to those in 10 normal people. RESULTS: No significant difference between two groups in the vocabulary identification and in the vocabulary identification from homophony (P>0.05) was observed (P>0.05). There was significant difference in the identification picture from vocabularies, the identification vocabulary from pictures and the vocabulary identification from sound (P>0.05). CONCLUSION: The patient with Broca aphasia is independent in the course of the orthographic input, and the ability to retrieve semantic information from orthographic activation word may not affected by phonological lexicon.     

Effect of external counterpulsation on asymmetry diethylarginine and endothelin levels in blood

AIM:To observe changes of serum asymmetry dimethylarginine (ADMA) and endothelin-1 (ET-1) levels,which reflect blood vessel endothelial function,after therapy of enhanced external counterpulsation(EECP) in patients who had taken percutaneous transluminal coronary angioplasty and stent.METHODS:Fifty one coronary heart disease patients (all of them had taken percutaneous tranluminal coronary angioplasty and stent) were distributed into two groups by matching them with ratio of 1∶2,17 patients in EECP group and 34 patients in control group.Both of two groups were given conventional medicine,in addition,EECP group was undertaken three courses of treatment of EECP.ADMA was detected by HPLC-fluorescence method,and ET-1 was detected by radio-immunity method.RESULTS:In EECP group ，compared with prior treatment,ADMA and ET-1 levels was obviously reduced (P<0.05).In control group,compared with prior treatment,ADMA had no significant difference (P>0.05) and ET-1 was higher (P<0.01).Compared between two groups,extent of decrease in ADMA and ET-1 levels in EECP group were more obvious than control group (P<0.01).Improvement of angina and decrease of frequency of angina was more obvious in EECP group than control group.Besides,decrease of ADMA level was positive correlation with them (r=0.85，0.70,respectively P<0.01).CONCLUSION:EECP reduces serum ADMA and ET-1 levels by increasing shear stress to vessel endothelium in coronary heart disease patients who had taken percutaneous tranluminal coronary angioplasty.It hints that EECP can improve endothelial function and provides experimental evidence for the combined treatment of coronary heart disease.     

罗非鱼无乳链球菌生物学特性与致病力研究

本试验分离纯化1株致使罗非鱼具有链球菌发病症状的病原菌,药敏试验结果表明,该菌对氯霉素等10种药物敏感。PCR扩增该菌的cfb基因,将扩增片段测序后与GenBank上登录的罗非鱼无乳链球菌的cfb基因序列进行比对。同时对该病原菌的生物学特性进行研究,包括培养时间和pH对该菌生长的影响;培养温度对其致病力的影响;探讨了不同攻毒方式(腹腔、胸腔和背部肌肉注射)与鱼发病快慢的关系;并对不同规格(100和10 g左右)的罗非鱼对该菌的易感性进行分析。试验结果表明,该病原菌为无乳链球菌(Streptococcus agalactiae);该菌的最适培养时间为11.5 h,最适培养pH为7.5;腹腔、胸腔和背部肌肉注射后,其发病时间几乎一致,但胸腔注射的试验鱼死亡速度最慢,背部肌肉注射是较安全可行的攻毒方式;33℃培养的无乳链球菌攻毒的罗非鱼发病最快,死亡最集中;100 g的试验鱼比10 g的试验鱼更易感染。     

副猪嗜血杆菌ompP2基因的克隆鉴定及序列分析

用PCR技术对华南地区分离的17株不同血清型副猪嗜血杆菌菌株ompP2基因进行克隆鉴定,并以CLUSTALS1和PHYLIP-3.68软件将ompP2基因序列进行比对和遗传进化分析.17株副猪嗜血杆菌茵株中均能扩出ompP2基因,克隆测序结果发现ompP2基因大小有所不同,与参考序列ABKM01000007的同源性在92％～99％之间.序列比较结果显示,ompP2基因与GenBank公布的副猪嗜血杆茵全基因组测序中的ompP2基因序列具有较高的同源性,不同菌株ompP2基因序列大小存在差异,为进一步验证ompP2蛋白的功能及相关研究奠定了基础.     

Sulodexide protects against hyperglycemia-induced retinal vascular injury via suppressing NOX4/NLRP3 pathway

AIM To investigate the effect of sulodexide (SDX) on high glucose-induced damage in retinal microvascular endothelial cells. METHODS (1) High-fat diet combined with intraperitoneal injection of streptozocin were used to induce type 2 diabetes mellitus (DM) followed by injection of saline or SDX in C57BL/6J male mice. Retinal microvascular leakage and density, and the protein levels of NLRP3 inflammasome-related proteins, zonula occludens-1 (ZO-1) and NADPH oxidase 4 (NOX4) were measured. (2) Human retinal microvascular endothelial cells (HRMECs) were treated with normal glucose or high glucose with or without SDX, and were further transfected with siRNA to knock down NOX4, or infected by adenovirus to over-express NOX4. The protein levels of ZO-1, VE-cadherin (VE-Cad), NOX4 and NLRP3 inflammasome-related proteins as well as the level of reactive oxygen species (ROS) were detected. RESULTS Treatment with SDX increased the protein level of ZO-1, attenuated retinal leakage and NLRP3 inflammasome activation, and enhanced the density of microvasculature and the number of ganglion cells in diabetic retinas. The protein levels of ZO-1 and VE-Cad were decreased, while the levels of NOX4, NLRP3 inflammasome-related proteins and ROS generation were increased in high glucose-treated HRMECs. Silencing of NOX4 inhibited high glucose-induced increases in NLRP3 inflammasome and ROS generation, and decreases in the protein levels of ZO-1 and VE-Cad. Over-expression of NOX4 significantly increased the levels of NLRP3 inflammasome-related proteins and ROS generation in HRMECs, and reduced the protein levels of ZO-1 and VE-Cad. Treatment with SDX partly reversed NOX4 over-expression-induced changes. CONCLUSION SDX alleviates hyperglycemia-induced retinal microvascular endothelial injury via inhibiting NOX4/ROS/NLRP3 pathways.     

Role of endoplasmic reticulum stress in trimethylamine N-oxide-induced oxidative stress in human umbilical vein endothelial cells

AIM: To investigate whether endoplasmic reticulum stress is involved in trimethylamine N-oxide (TMAO)-mediated oxidative stress in human umbilical vein endothelial cells (HUVECs). METHODS: The cell viability was examined by CCK-8 assay. The cells were stained by DCFH-DA, and the intracellular level of reactive oxygen species (ROS) was observed by phase-contrast microscopy and detected by flow cytometric analysis. The protein levels of phospho-IRE-1α, IRE-1α and GRP78/BiP were detected by Western blot. RESULTS: TMAO exerted no significant effect on the viability of HUVECs. For a long period (>24 h), even a low concentration (10 μmol/L) of TMAO increased the oxidative stress level in the HUVECs (P<0.05). TMAO increased the phosphorylation level of IRE-1α and significantly up-regulated the protein level of GRP78/BiP in HUVECs (P<0.01). Pretreatment with STF-083010, an inhibitor of IRE1α, for 1 h reduced TMAO-induced oxidative stress in HUVECs (P<0.05). CONCLUSION: Endoplasmic reticulum stress is involved in TMAO-induced oxidative stress in HUVECs.     

南沙群岛海域春、秋季浮游介形类生态学特征

为了解南沙群岛海域春、秋季浮游介形类生态学特征,根据2013年春季(3月中旬至4月初)及秋季(9月)在南沙群岛开展的两个航次生态环境综合调查资料,对该海域浮游介形类生态学特征进行分析研究。结果显示:(1)南沙群岛海域浮游介形类物种组成较为丰富,记录 45种隶属于2科、4亚科、19属。其中,春季有21种,秋季38种,两季均出现的共有种有14个;(2)春、秋两季浮游介形类优势种差异明显,春季优势种组成较为单一,数量低,主要生态类群为外海暖水类群和广温暖水类群,秋季优势种组成较为复杂,数量多,主要为外海暖水类群。春季优势种由后圆真浮萤、葱萤、尖头毛浮萤组成,秋季由纳米海萤、双叉真浮萤、刺喙葱萤、短形小浮萤、粗大后浮萤组成。研究表明:(1)春秋两季浮游介形类密度差异显著,密度分布不均,局部区域密度较高,与该海域局部环流密切相关;(2)浮游介形类两季平均丰富度、多样性指数和均匀度分别为0.59、1.37和0.55,秋季高于春季;(3)春季及秋季均分为岛礁群落和外海群落,群落分布随洋流改变呈季节性变化;(4)受季风影响的大环流及局部环流是浮游介形类种类组成、数量分布及多样性呈明显季节差异的主要因素。     

Caelastrol and pristimerin derived from Leigongteng inhibit promoter activity of HLA-B*2705 gene

AIM: To screen the effective chemicals, which can suppress the promoter activity of the HLA-B*2705 gene as potential therapeutic agents. METHODS: The HeLa-HLA-B27, 293T-HLA-B27 stable transfectants were used to monitor the effect of 12 264 chemicals through high throughput screening (HTS). Chemicals which regulates HLA-B*2705 promoter activity more than 150% or less than 60% were picked out for the further IC50/EC50 and cell viability detection. RESULTS: (1) The primary screening used by 293T-HLA-B27 stable transfectant yielded about 5.1% hits which either suppressed (556 chemicals) or enhanced (68 chemicals) the HLA-B*2705 promoter activity. (2) A reconfirmation screening was carried out with these 624 of the candidates using transfected HeLa-HLA-B27 cells. Seventy hits were confirmed. (3) Based on the bioinformatics of those positive hits, 40 chemicals were selected for in-depth analysis by dose-response experiment and IC50/EC50 detection. Six suppressors showed potential pharmacological activities. Interestingly, two suppressors (celastrol and pristimerin) are derived from Leigongteng, a herbal medicine already used for several decades for treatment of immune regulatory and inflammatory diseases. Four active chemicals were computer designed with no relevance to the above structures. CONCLUSION: Chinese traditional herb Nansheteng and Leigongteng might be the potential drugs for HLA-B27 positive patients. These results provide new direction for research in both the therapeutics and the pathogenesis of spondyloarthritis.