Development and Primary Application of an Indirect ELISA Based on Rep Protein to Analyze Antibodies against Porcine Cocirvirus-like Virus (PCLV)

Porcine circovirus-like virus (PCLV) is a member of circovirus that contains a single-strand DNA genome, which may be one of the pathogens that causes diarrheal symptoms in pigs. The Rep protein encoded by the genome of PCLV may be responsible for viral genome replication. The development of serological detection methods for PCLV is of great necessity for clinical diagnosis, as well as epidemiological investigations. Therefore, this study attempted to build an indirect enzyme-linked immunosorbent assay (ELISA) to examine antibodies against PCLV based on the His-tagged recombinant Rep protein. Full-length PCLV Rep protein was induced and expressed in E. coli and was purified as an antigen to establish an ELISA detection kit. The purified Rep protein was used to inject into mice to produce specific antibodies. There was no cross-reaction of Rep-based ELISA with antisera against other porcine viruses. The intra-assay and inter-assay coefficient variations (CVs) were 0.644–8.211% and 0.859–7.246%, respectively, indicating good repeatability. The non-cross-reaction with TGEV, PRRSV and PCV2 testing showed high sensitivity and high specificity for this ELISA assay. A total of 1593 serum samples collected from different pig farms in Jiangxi Province were tested for anti-PCLV Rep antibodies, and 284 (17.83%) of the 1593 samples were Rep antibody positive. Altogether, the indirect ELISA detection tool developed in this study could be applied to examine serum of PCLV antibodies with good repeatability, high sensitivity and high specificity. In addition, field sample detection results suggested that the PCLV antibody has a low prevalence in pig populations in Jiangxi Province of China.     

Role of 5-lipoxygenase pathway in the regulation of RAW 264.7 macrophage proliferation

Arachidonic acid (AA) metabolites control cell proliferation, among other physiologic functions. RAW 264.7 macrophages can metabolise AA through the cyclooxygenase and lipoxygenase (LOX) pathways. We aimed to study the role of AA-metabolites derived from 5-LOX in the control of RAW 264.7 macrophage growth. Our results show that zileuton, a specific 5-LOX inhibitor, and nordihydroguaiaretic acid (NDGA), a non-specific LOX inhibitor, inhibit cell proliferation and [(3)H]-thymidine incorporation in a concentration-dependent fashion. Growth inhibition induced by NDGA can be explained by an apoptotic process, while zileuton does not seem to induce apoptosis. Moreover, these treatments delay the cell cycle, as analysed by flow cytometry. On the other hand, the leukotriene (LT) B(4) receptor antagonist U-75302, the LTD(4) receptor antagonists LY-171883 and MK-571, and the cysteinyl-LT receptor antagonist REV-5901 also inhibit cell proliferation and [(3)H]-thymidine incorporation in a concentration-dependent manner, and delay the RAW 264.7 cell cycle. However, these antagonists did not induce annexin V staining, caspase activation or DNA fragmentation. Furthermore, we demonstrated that exogenous addition of LTB(4) or LTD(4) revert the cell growth inhibition induced by zileuton or the leukotriene receptor antagonists mentioned above. Finally, we observed that LTB(4) and LTD(4), in the absence of growth factors, have pro-proliferative effects on macrophages, and we obtained preliminary evidences that this effect could be through mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways. In conclusion, our results show that the interaction between LTB(4) and LTD(4) with its respective receptor is involved in the control of RAW 264.7 macrophage growth.     

Smoking Exposure Is Associated with Serum Vitamin D Deficiency in Children: Evidence from the Japan Environment and Children’s Study

Tobacco smoke exposure is known to lower serum 25-hydroxyvitamin D (25(OH)D) concentrations. This study evaluated the association between passive smoking and vitamin D deficiency (VDD) in young children using data from the Japan Environment and Children’s Study (JECS), the largest birth cohort study in Japan. Information on parental smoking status was extracted from a survey of JECS for children aged 1.5 years and data for serum 25(OH)D concentrations were obtained from blood tests in the Sub-Cohort Study of JECS performed at age 2 years. Logistic regression and linear models were fitted to evaluate the association between these variables. Data were analyzed for 4593 children. After adjusting for covariates, smoke exposure was significantly associated with increased incidence of VDD (OR 1.35; 95% CI, 1.14–1.59) according to the logistic model. The linear model indicated that passive smoking negatively predicted de-seasonalized serum 25(OH)D concentrations (β −0.5; 95% CI −0.95 to −0.08) in children aged 2 years. The results suggest that smoke exposure is a risk factor for VDD in children. Given that VD plays a crucial role in bone metabolism and the immune system, our findings are significant for clinical and public health.     

Sotalol and metoprolol comparison of their anti-hypertensive effect

Summary 28 patients, aged 35–62 years, with uncomplicated hypertension, entered a double-blind, crossover study, in which the effects of single daily doses of sotalol and metoprolol were compared. Both drugs exerted a clinically useful anti-hypertensive effect as monotherapy, or in combination with a thiazide diuretic. No significant difference in hypotensive effects was noted between the two beta-blocking agents, when the dose was titrated to an optimal clinical effect. Treatment with sotalol and metoprolol was associated with a clinically insignificant increase in serum uric acid concentration. The side-effects observed were few, and in only two cases was therapy discontinued. We regard both sotalol and metoprolol as useful anti-hypertensive drugs.     

Ionized and total magnesium concentration in patients with severe preeclampsia-eclampsia undergoing magnesium sulfate therapy

AIM: As ionized magnesium is the active form of magnesium and exerts a therapeutic effect, the present study was performed to determine the levels and correlations between ionized and total magnesium under baseline and therapeutic conditions in patients with severe preeclampsia and eclampsia receiving magnesium sulfate. METHODS: Fifty singleton patients with severe preeclampsia received a loading dose of 4 g of magnesium sulfate, followed by 2 g per hour as maintenance dose until 24 h after delivery, or 24 h after the last seizure in case of postpartum convulsions. Serial blood samples were taken before magnesium sulfate infusion, 30 min and 240 min after the initiation of the infusion and 4 h after the discontinuation of the drug. Data were analyzed by repeated measure ANOVA and paired t-test. RESULTS: Baseline levels of total and ionized magnesium were 2.4+/-0.6 mEq/L and 1.3+/-0.5 mEq/L (mean+/-SD), respectively. Putative level of 4 mEq/L of total magnesium was not obtained in up to 42% of patients during the treatment. There was not any significant correlation between the two forms of magnesium under baseline and therapeutic conditions. CONCLUSION: Despite the effectiveness of the standard regimen of magnesium sulfate in the treatment and prevention of eclamptic seizures, it can not provide the proposed therapeutic level of magnesium in all patients. With respect to the lack of correlation between ionized and total magnesium, further studies are necessary to investigate the superiority of measurement of ionized, rather than total magnesium, for titration of therapeutic magnesium sulfate infusion.     

2型糖尿病患者血尿酸水平与尿白蛋白排泄率及颈动脉内中膜厚度的关系

目的 探讨2型糖尿病(T2DM)患者血清尿酸(SUA)水平与尿白蛋白排泄率(UAER)和颈动脉内中膜厚度(CIMT)的关系.方法 2008年5月-2009年5月收治T2DM患者372例,其中男性184例,女性188例.收集相关临床资料.Pearson法检验患者SUA水平与各项临床指标的相关性;多元逐步回归法分析影响UAER和CIMT的相关因素.结果 男性T2DM患者SUA水平明显高于女性(P<0.01).男性和女性T2DM患者SUA水平均与UAER相关;在校正内生肌酐清除率(Ccr)的影响后,相关性仍然存在(男性:r=0.24, P<0.01;女性:r=0.29, P<0.01).女性T2DM患者SUA水平与CIMT相关(r=0.29, P<0.01). 多元逐步回归分析显示,SUA是T2DM患者UAER(男性: β=0.16,P<0.05;女性: β=0.20,P<0.05)及女性T2DM患者CIMT的独立相关因素(β=0.16,P<0.05).结论 SUA在T2DM患者肾病和心血管病的发生和发展中起一定作用,控制SUA水平可能有助于延缓糖尿病肾病和防治糖尿病血管并发症.     

白头翁提取液对溃疡性结肠炎模型大鼠血清TNF-α、IL-10的影响

目的 研究白头翁提取液对溃疡性结肠炎模型大鼠血清细胞因子TNF-a、IL-10的影响.方法 将Wistar大鼠48只随机分为6组:正常对照组、模型组、中药高、中、低剂量组、SASP组,采用TNBS法制备溃疡性结肠炎大鼠模型,中药高、中、低各剂量组分别按相应剂量给予白头翁水提取液灌胃;SASP组给予SASP水溶液灌胃;正常对照组和模型组给予等量生理盐水灌胃,各组均连续给药15 d后观察大鼠结肠黏膜损伤指数(CM-DI)及检测血清中TNF-α、IL-10含量.结果 与正常对照组比较,模型组大鼠CMDI评分均增加;与模型组比较,白头翁各剂量组与SASP组CMDI评分均降低;与SASP组比较,白头翁中剂量组显著降低.与正常对照组相比,模型组大鼠血清中TNF-α含量显著上升,IL-10含量显著下降;与模型组相比,白头翁各剂量组与SASP组大鼠血清中TNF-α含量均显著下降,IL-10含量显著上升.结论 白头翁水提取液在溃疡性结肠炎的治疗中可能通过降低血浆中TNF-α水平和升高IL-10水平,起到促进溃疡性结肠炎肠黏膜修复与溃疡愈合的作用.