Thrombin impairs the angiogenic activity of extravillous trophoblast cells via monocyte chemotactic protein-1 (MCP-1): A possible link with preeclampsia

Cytokines’ secretion from the decidua and trophoblast cells has been known to regulate trophoblast cell functions, such as Extravillous trophoblasts (EVTs) cell migration and invasion and remodeling of spiral arteries. Defective angiogenesis and spiral arteries transformation are mainly caused by proinflammatory cytokines and excessive thrombin generation during preeclampsia. Monocyte chemotactic protein-1 (MCP-1), a crucial cytokine, has a role in maintaining normal pregnancy. In this study, we explored whether thrombin regulates the secretion of MCP-1 in HTR-8/SVneo cells; if yes, what is its function? We used HTR-8/SVneo cells, developed from ?rst trimester villous explants of early pregnancy, as the model of EVTs. MCP-1 gene silencing was performed using gene-specific siRNA. qPCR and ELISA were performed to estimate the expression and secretion of MCP-1. Here, we found that thrombin enhanced the secretion of MCP-1 in HTR-8/SVneo cells. Proteinase-activated receptor-1 (PAR-1) was found as the primary receptor, regulating MCP-1 secretion in these cells. Furthermore, MCP-1 secretion is modulated via protein kinase C (PKC) α, β, and Rho/Rho-kinase-dependent pathways. Thrombin negatively regulates HTR-8/SVneo cells’ ability to mimic tube formation in an MCP-1 dependent manner. In conclusion, we propose that thrombin-controlled MCP-1 secretion may play an essential role in normal placental development and successful pregnancy maintenance. Improper thrombin production and MCP-1 secretion during pregnancy might cause inadequate vascular formation and transformation of spiral arteries, which may contribute to pregnancy disorders, such as preeclampsia.     

新疆猪粪便戊型肝炎病毒RNA的检测及序列分析

从戊型肝炎病毒(Hepatitis Evirus,HEV)IgG检测阳性的新疆某猪场采集70份猪粪便,利用逆转录套式聚合酶链方法(RT-nPCR),检测HEV RNA,其中13份为阳性,阳性率18．57％。将PCR扩增产物克隆到pMD18-T载体上,构建成重组质粒并测序,结果表明,13株猪源HEV分离株在HEV ORF2 348bp核苷酸序列的同源性为97．1％～100％,为同一基因型;与HEVⅠ、Ⅱ、Ⅲ、Ⅳ的同源性分别为74．1％-77．6％,71．6％-74．1％,73.3％～78．2％和82．8％-91.4％,与ⅣA亚型的同源性同源性最高达89．4％-91．4％。以该核苷酸片段绘制的基因进化树显示13株猪源HEV与HEV Ⅳ T1株在同一分支上,属基因Ⅳ型;与国内其他猪源HEV分离株该片段核苷酸序列的同源性为82．6％-91．3％,提示中国猪源HEV的基因型比较一致,同属HEV Ⅳ型。     

新疆猪粪便戊型肝炎病毒RNA的检测及序列分析

从戊型肝炎病毒(Hepatitis E virus,HEV)IgG检测阳性的新疆某猪场采集70份猪粪便,利用逆转录套式聚合酶链方法(RT-nPCR),检测HEV RNA,其中13份为阳性,阳性率18.57%.将PCR扩增产物克隆到pMD18-T载体上,构建成重组质粒并测序,结果表明,13株猪源HEV分离株在HEV ORF2 348bp核苷酸序列的同源性为97.1%～100%,为同一基因型;与HEV Ⅰ、Ⅱ、Ⅲ、Ⅳ的同源性分别为74.1%～77.6%,71.6%～74.1%,73.3%～78.2%和82.8%～91.4%,与ⅣA亚型的同源性同源性最高达89.4%～91.4%.以该核苷酸片段绘制的基因进化树显示13株猪源HEV与HEVⅣT1株在同一分支上,属基因Ⅳ型;与国内其他猪源HEV分离株该片段核苷酸序列的同源性为82.6%～91.3%,提示中国猪源HEV的基因型比较一致,同属HEVⅣ型.     

用反转录—聚合酶链反应检测B组轮状病毒

轮状病毒（Ｒｏｔａｖｉｒｕｓ）是属于呼肠病毒科（Ｒｅｏｖｉｒｉｄａｅ）的双链ＲＮＡ（ｄｓＲＮＡ）病毒。至今已将轮状病毒分为七个组（Ａ～Ｇ）。已经发现的Ｂ组轮状病毒分别来自人、大鼠、牛、猪、羊。近十年来,通过轮状病毒的研究,轮状病毒Ｂ组已被公认为引起人...     

Synthetic LXR agonist inhibits the development of atherosclerosis in New Zealand White rabbits

The nuclear receptors Liver X receptors, LXRα and LXRβ, regulate cholesterol and triglyceride metabolism. We and others have previously reported that synthetic LXR agonists reduced atherosclerosis in models of mouse with no detectable plasma cholesteryl ester transfer protein (CETP) activity, which plays an important role in reverse cholesterol transport. In the present study, we investigated the effect of LXR activation in rabbits to elucidate the influence of CETP activity. First, we cloned rabbit LXRs cDNA. The data indicated that rabbit LXRα was mostly highly expressed in the liver, whereas LXRβ expression was ubiquitous. Next, we investigated the effect of LXR agonist on lipid levels. Treatment with LXR agonist T0901317 increased plasma CETP activity and consequently elevated LDL, but no change in HDL. High cholesterol (HC) diet-feeding, which is thought to provide oxysterols as the natural agonists, could also increase expression of CETP and other LXR target genes. Finally, we tested T0901317 in the atherosclerosis intervention study. Chronic administration of T0901317 significantly reduced atherosclerosis in HC diet-fed rabbits despite less favorable lipid profiles, i.e. increases of plasma triglycerides and no change of HDL. T0901317 induced ATP-binding cassette transporters ABCA1 and ABCG1 and suppressed inflammatory genes expression in the aorta, suggesting that direct actions of LXR agonist on vascular gene expression are likely to contribute to the antiatherogenic effect. The present work strongly supports the idea that LXR agonists could be beneficial as therapeutic agents for treatment of atherosclerosis.     

应用RT-PCR制备登革病毒诊断基因芯片探针

根据GenBank数据库中的生物信息,利用BLAST免费分析软件找出4种型别登革病毒的保守序列及各型特异性序列,针对上述序列设计引物经RT-PCR扩增登革病毒的特异片段,利用此RT-PCR法收集探针是一种快速、简便制备基因芯片探针的实用方法。     

Evidence of absorption rate dependency of ibuprofen inversion in the rat

Ibuprofen (IB) is a chiral 2-arylpropionic acid derivative used as a nonsteroidal antiinflammatory drug (NSAID). It undergoes substantial R to S chiral inversion in humans and rats. In addition to systemic inversion, presystemic chiral inversion has been suggested for IB in humans but only after administration of formulations with slow absorption rates. In search for a suitable animal model, the absorption rate dependency of the extent of inversion was examined in male Sprague–Dawley rats given 20 mg/kg of racemic IB in aqueous solution (T_max, 0.6 h), suspension (T_max, 1 h) or as sustained release granules (T_max, 2.3 h). In addition, (R)-IB (5 mg/liter) was incubated in the presence of everted rat gut segments in an organ bath at 37°. After sustained release granules, the S:R AUC ratios (7.3 ± 1.5) were significantly higher than suspension (3.6 ± 1.1) and solution (3.5 ± 0.2). Accordingly, AUC_S and AUC_R, as percent of the total AUC (S + R), significantly increased and decreased, respectively, after administration of the sustained released granules as compared with the solution and suspension. A significant positive linear correlation was found between the S:R AUC ratios and the corresponding T_max for (R)-IB (r = 0.82). In vitro, (R)-IB was inverted by everted jejunum (12.2 ± 1.6%), ileum (14.2 ± 2.0%), and colon (4.4 ± 0.6%) segments. IB was also glucuronidated in the presence of the intestinal segments. Therefore, similar to earlier observations made in humans, in the rat, the S:R AUC ratio was positively and significantly correlated with the absorption rate from the dosage form. Rat small intestine was capable of inverting and conjugating (R)-IB. Hence, rat is a suitable model for studying the chiral inversion of IB. © 1994 Wiley-Liss, Inc.     

Chibby functions in Xenopus ciliary assembly,embryonic development,and the regulation of gene expression

Wnt signaling and ciliogenesis are core features of embryonic development in a range of metazoans. Chibby (Cby), a basal-body associated protein, regulates β-catenin-mediated Wnt signaling in the mouse but not Drosophila. Here we present an analysis of Cby?s embryonic expression and morphant phenotypes in Xenopus laevis. Cby RNA is supplied maternally, negatively regulated by Snail2 but not Twist1, preferentially expressed in the neuroectoderm, and regulates β-catenin-mediated gene expression. Reducing Cby levels reduced the density of multiciliated cells, the number of basal bodies per multiciliated cell, and the numbers of neural tube primary cilia; it also led to abnormal development of the neural crest, central nervous system, and pronephros, all defects that were rescued by a Cby-GFP chimera. Reduction of Cby led to an increase in Wnt8a and decreases in Gli2, Gli3, and Shh RNA levels. Many, but not all, morphant phenotypes were significantly reversed by the Wnt inhibitor SFRP2. These observations extend our understanding of Cby?s role in mediating the network of interactions between ciliogenesis, signaling systems and tissue patterning.     

反义trxs基因对转基因小麦种子内源trxh基因表达的影响

以转反义硫氧还蛋白基因(anti-trxs)株系01TY70-1-17-5及其对照小麦品种‘豫麦70’为试验材料,以小麦中稳定表达的肌动蛋白基因actin为内标,用半定量反转录聚合酶链式反应(semi-QRT-PCR)方法,对转基因株系及其对照种子中trxh基因时空表达情况进行了检测。检测结果表明,花后15~30d转基因株系trxh基因转录量平均比对照下降了20.1%,花后25d显著低于对照(P<0.05);胚乳trxh基因转录量最低,平均比对照低19.4%;种子吸涨24h时间内,转基因株系trxh基因转录量较对照均略低,但差异不显著。表明,外源trxs基因的导入直接干扰了内源基因的表达。     

Suppression of major histocompatibility complex class I and class II gene expression in Listeria monocytogenes-infected murine macrophages

Macrophage cells play a central role during infection with Listeria monocytogenes by both providing a major habitat for bacterial multiplication and presenting bacterial antigens to the immune system. In this study, we investigated the influence of L. monocytogenes infection on the expression of MHC class I and class II genes in two murine macrophage cell lines. Steady-state levels of I-Aβ chain mRNA were decreased in both resting J774A.1 and P388D₁ macrophages infected with L. monocytogenes whereas reduction of H-2K mRNA was only observed in P388D₁ cells. In addition, L. monocytogenes suppressed induction of MHC class I and class II mRNAs in response to γ-interferon as well as the maintenance of the induced state in activated P388D₁ macrophages. Exposure to the non-pathogenic species L. innocua or a deletion mutant of L. monocytogenes, which lacks the lecithinase operon, did not cause a reduction in H-2K and I-Aβ mRNA levels nor suppress expression of Ia antigens. Inhibition of MHC gene expression may represent an important part of the cross-talk between L. monocytogenes and the macrophage that probably influences the efficiency of a T cell-mediated immune response and thus the outcome of a listerial infection.     

PCR方法检测水稻中存在G蛋白基因家族

ＰＣＲ方法检测水稻中存在Ｇ蛋白基因家族陈忠英,王钧（中国科学院上海植物生理研究所,２００００３２）关键词Ｇ蛋白;聚合酶链式反应（ＰＣＲ）;水稻苗;ＤＮＡ序列分析植物细胞对各种外源和内源的刺激,如光、重力、病原、激素等都有灵敏复杂的反应,但对植物信息传...     

纳米金对PCR扩增效率影响的机制探讨

近年来,纳米金粒子对聚合酶链式反应（polymerasechainreaction,PCR）的增效作用倍受关注,但是其具体的机制仍未明确提出。研究发现在PCR反应中,纳米粒子的增效作用是存在最佳浓度的,增加DNA聚合酶或者小牛血清蛋白（BSA）可以消除纳米金粒子导致的抑制。我们认为,纳米金粒子可能起到了类似于聚合酶B亚基的作用,提高了DNA聚合酶的延伸能力;而过量纳米金对PCR的抑制作用可能与纳米金结合单链DNA产生的位阻效应有关。