AFM对黄精赞育胶囊优选方处理前后大鼠弱精子超微结构的观察研究

目的:应用原子力显微镜技术实现对大鼠精子超微结构的实时成像,对比观察中药作用前后弱精子超微结构的变化.方法:制作少弱精症大鼠模型;运用非接触模式原子力显微镜技术对比观察正常精子和病理性精子在中药作用前后超微结构的动态变化.结果:获得精子头体、颈部和鞭毛等部位的实时超微结构图像.结论:AFM可直接探测精子头体的各种畸形,实现对精子全貌的观测.通过修复弱精子超微结构的病理形态学缺陷可能是黄精赞育胶囊优选方改善弱精子质量的机制之一.     

实时定量PCR方法检测鼻咽癌病人全血EB病毒拷贝数的实验研究

EBV is detected in more and more tumors, and is relative to carcinogenesis. We studied the copies of EBV DNA in whole blood of NPC patients and healthy controls by real-time quantitative PCR. In the 73 NPC patients and 83 controls, the positive rate of EBV in blood of NPC is 46.6%, while 13.3% in control. The mean copy number is 3.9 x 10(4) copys/microgram DNA in controls, which is much higher than NPC patients (which is 1.7 x 10(5) copies/microgram DNA). EBV infection is relative with NPC, while the lytic form of EBV maybe more important than its latent form. These results suggest that whole blood EBV DNA may be a valuable tool for molecular diagnosis of NPC.     

Bovine colostrum promoted ileal health in newborn lambs at 24 h after birth: insight from intestinal morphology and innate immunity

The contribution of colostrum to passive immunity transfer and intestinal protection in newborn ruminants is well known; however, it is currently unclear how colostrum intake affects intestinal innate immunity. We investigated the effects of bovine colostrum intake on ileal morphology, expression of genes involved in intestinal innate immunity, and serum concentrations of inflammatory cytokines in newborn lambs. Twenty-seven newborn male Hu lambs were used, of which 18 were bottle-fed either bovine colostrum (C24h; n = 9) or bovine mature milk (M24h; n = 9) within the first 2 h after birth at an intake of approximately 8% of BW; the remaining nine lambs did not receive any feeding (N24h). Blood and ileal tissue samples were collected after the lambs were slaughtered at 24 h after birth. Ileal villus height and villus height-to-crypt depth ratio were significantly higher in C24h than those in N24h and M24h lambs (P < 0.01). Messenger RNA (mRNA) abundance of toll-like receptor (TLR)-2, TLR3, TLR4, TLR6, TLR7, TLR8 and tumour necrosis factor alpha in the ileum was lower in C24h than that in N24h lambs (P < 0.05). Moreover, C24h lambs had a lower TLR3 mRNA abundance (P < 0.01) and a trend of lower TLR6 (P = 0.06) and interleukin 1 beta (P = 0.08) expression compared with those in M24h lambs. We also observed strong positive correlations of tumour necrosis factor alpha expression with that of TLR2 (r = 0.71; P < 0.001), TLR4 (r = 0.88; P < 0.001) and TLR8 (r = 0.83; P < 0.001). Interestingly, the expression of barrier-related molecules, including mucin-13, lysozyme, claudin (CLDN)-1, CLDN2, CLDN4, CLDN7, CLDN12, occludin, zonula occluden-1 and junctional adhesion molecule-1, was consistently lower in C24h lambs than that in N24h and M24h lambs (P < 0.05). These results indicated that the beneficial roles of colostrum intake on intestinal protection in newborn lambs were associated with low TLR expression, which was reflected by improved intestinal development and reduced inflammatory response. Further studies using fluorescence in situ hybridisation and immunohistochemical methods are needed to further explore the mechanisms underlying the lower expression of intestinal barrier-related molecules due to colostrum feeding.     

DNA-methylation-mediated silencing of miR-7-5p promotes gastric cancer stem cell invasion via increasing Smo and Hes1

Cancer stem cells are undifferentiated cancer cells that have self-renewal ability, a high tumorigenic activity, and a multilineage differentiation potential. MicroRNAs play a critical role in regulating gene expression during carcinogenesis. Here, we investigated the role of miR-7 and the mechanism by which it is dysregulated in gastric cancer stem cells (GCSCs). The stem cell marker, CD44, was used to sort GCSCs by fluorescence-activated cell sorting. We found that CD44 (+) cells have higher invasiveness and form more number of sphere colonies than CD44 (−) cells. Quantitative real-time polymerase chain reaction (PCR) revealed that the miR-7-5p expression was remarkably downregulated in GCSCs but was significantly increased in the methionine-deprived medium. The downregulation of miR-7-5p results from the increased DNA methylation in the promoter region using the methylation-specific PCR. Overexpression of miR-7-5p reduced the formation of colony and decreased the invasion of GCSCs through targeting Smo and Hes1 and subsequent repressing Notch and Hedgehog signaling pathways in vitro. Notably, upregulating miR-7-5p inhibited the growth of tumor in the xenograft model. Hence, these data demonstrated that miR-7-5p represses GCSC invasion through inhibition of Smo and Hes1, which provides a potential therapeutic target of gastric cancer treatment.     

激素联合丙种球蛋白治疗小儿重症肌无力的疗效及对患儿免疫球蛋白和补体的影响

目的：探讨激素联合丙种球蛋白治疗小儿重症肌无力的临床疗效及对患儿免疫球蛋白和补体的影响。方法：回顾性分析在我院治疗的70例重症肌无力患儿的临床资料,采用随机序号的方式将其分为观察组和对照组各35例,观察组给予甲泼尼龙联合丙种球蛋白,对照组仅给予甲泼尼龙,观察两组的临床疗效及免疫球蛋白和补体变化情况。结果：观察组总有效率为94．3％明显优于对照组74．3％,两组比较有统计学意义（P〈0．05）;观察组症状明显缓解时间（6．55±1．35）d以及总住院天数（17．15±3．65）d较对照组明显缩短,两组比较差异均有统计学意义（P〈0．05）。结论：采用激素联合丙种球蛋白治疗小儿重症肌无力,可以明显改善患者肌无力症状,获得较为满意的临床疗效,值得进一步推广使用。     

Lineage‐specific plastid degradation in subtribe Gentianinae (Gentianaceae)

The structure and sequence of plastid genomes is highly conserved across most land plants, except for a minority of lineages that show gene loss and genome degradation. Understanding the early stages of plastome degradation may provide crucial insights into the repeatability and predictability of genomic evolutionary trends. We investigated these trends in subtribe Gentianinae of the Gentianaceae, which encompasses ca. 450 species distributed around the world, particularly in alpine and subalpine environments. We sequenced, assembled, and annotated the plastomes of 41 species, representing all six genera in subtribe Gentianinae and all main sections of the species‐rich genus Gentiana L. We reconstructed the phylogeny, estimated divergence times, investigated the phylogenetic distribution of putative gene losses, and related these to substitution rate shifts and species’ habitats. We obtained a strongly supported topology consistent with earlier studies, with all six genera in Gentianinae recovered as monophyletic and all main sections of Gentiana having full support. While closely related species have very similar plastomes in terms of size and structure, independent gene losses, particularly of the ndh complex, have occurred in multiple clades across the phylogeny. Gene loss was usually associated with a shift in the boundaries of the small single‐copy and inverted repeat regions. Substitution rates were variable between clades, with evidence for both elevated and decelerated rate shifts. Independent lineage‐specific loss of ndh genes occurred at a wide range of times, from Eocene to Pliocene. Our study illustrates that diverse degradation patterns shape the evolution of the plastid in this species‐rich plant group.     

Cathepsin S deficiency results in abnormal accumulation of autophagosomes in macrophages and enhances Ang II-induced cardiac inflammation

Background

Cathepsin S (Cat S) is overexpressed in human atherosclerotic and aneurysmal tissues and may contributes to degradation of extracellular matrix, especially elastin, in inflammatory diseases. We aimed to define the role of Cat S in cardiac inflammation and fibrosis induced by angiotensin II (Ang II) in mice.

Methods and Results

Cat S-knockout (Cat S^−/−) and littermate wild-type (WT) C57BL/6J mice were infused continuously with Ang II (750 ng/kg/min) or saline for 7 days. Cat S^−/− mice showed severe cardiac fibrosis, including elevated expression of collagen I and α-smooth muscle actin (α-SMA), as compared with WT mice. Moreover, macrophage infiltration and expression of inflammatory cytokines (tumor necrosis factor α, transforming growth factor β and interleukin 1β) were significantly greater in Cat S^−/− than WT hearts. These Ang II-induced effects in Cat S^−/− mouse hearts was associated with abnormal accumulation of autophagosomes and reduced clearance of damaged mitochondria, which led to increased levels of reactive oxygen species (ROS) and activation of nuclear factor-kappa B (NF-κB) in macrophages.

Conclusion

Cat S in lysosomes is essential for mitophagy processing in macrophages, deficiency in Cat S can increase damaged mitochondria and elevate ROS levels and NF-κB activity in hypertensive mice, so it regulates cardiac inflammation and fibrosis.     

竹节参鲨烯环氧酶基因的克隆与生物信息学分析

根据已报道的植物鲨烯环氧酶(SE)基因特异性引物克隆竹节参SE基因.结果表明,克隆得到竹节参SE全长为1 632 bp,编码539个氨基酸,命名为PjSE.生物信息学分析指出,PjSE基因的氨基酸序列与人参属植物人参、三七、越南人参的同源性依次为99％、98％和98％.推测PjSE基因定位于线粒体(mTP)、叶绿体(cTP)和分泌途径(SP).PjSE基因存在保守结构域FAD结合位点,含4个跨膜结构域和7个motif结构位点.PjSE蛋白二级结构以无规则卷曲(Random coli)和α-螺旋(Alpha helix)为主要结构元件,延伸链(Extended strand)和β转角(Beta turn)分散于整个蛋白质中,百分比依次占40.82％、37.48％、15.77％、5.94％;该蛋白能折叠形成典型的三维结构.     

Autophagy in plants: Physiological roles and post‐translational regulation

In eukaryotes, autophagy helps maintain cellular homeostasis by degrading and recycling cytoplasmic materials via a tightly regulated pathway.Over the past few decades, significant progress has been made towards understanding the physiological functions and molecular regulation of autophagy in plant cells. Increasing evidence indicates that autophagy is essential for plant responses to several developmental and environmental cues, functioning in diverse processes such as senescence, male fertility, root meristem maintenance, responses to nutrient starvation,and biotic and abiotic stress. Recent studies have demonstrated that, similar to nonplant systems,the modulation of core proteins in the plant autophagy machinery by posttranslational modifications such as phosphorylation, ubiquitination,lipidation, S-sulfhydration, S-nitrosylation, and acetylation is widely involved in the initiation and progression of autophagy. Here, we provide an overview of the physiological roles and posttranslational regulation of autophagy in plants.