黄芪汤对延缓C离子辐射致大鼠肾组织细胞凋亡的影响及相关机制

目的: 探讨黄芪汤抑制¹²C⁶⁺离子辐射脑模型鼠肾组织细胞凋亡的分子保护机制。方法: 50只SPF级Wistar大鼠随机分为正常对照组,单纯辐射模型组,黄芪汤(高、中、低剂量)组。正常对照组和单纯辐射模型组给予等体积生理盐水灌胃10 ml/(kg·d),黄芪汤治疗组分别灌胃给予黄芪汤18、9、4.5 g/(kg·d),连续给药2周。7 d后除正常对照组外,其余各组大鼠脑组织给予4Gy ¹²C⁶⁺离子束单次照射,辐射后第7日处死各组大鼠。HE染色法观察大鼠肾脏的病理形态变化,ELISA法检测大鼠血清IL-6的含量,实时荧光定量PCR法测定大鼠肾脏Bcl-2、Bax和Caspase-3的基因表达,免疫组化法检测大鼠肾脏Bcl-2、Bax、Caspase-3和NF-κB的蛋白表达。结果: 与正常对照组比较,单纯辐射模型组体重和肾脏指数均显著降低,血清IL-6的含量显著升高,肾脏Bcl-2的基因表达和蛋白表达均显著降低,Bax和Caspase-3的基因表达和蛋白表达均显著升高,NF-κB的蛋白表达也显著升高(P＜ 0.01),单纯辐射组肾小球系膜细胞明显增生,肾小管间质血管明显扩张充血,肾小管管腔狭窄、不规则。与单纯辐射模型组相比,黄芪汤高剂量组体重和肾脏指数均明显升高,黄芪汤各干预组肾脏Bcl-2的基因表达和蛋白表达均显著升高(P＜0.05或P＜0.01);而黄芪汤中、高剂量组Bax和Caspase-3的蛋白表达均显著降低,各干预组血清IL-6的含量显著降低,肾脏Bax和Caspase-3的基因表达均显著降低,肾脏NF-κB的蛋白表达显著下降(P＜0.05或P＜0.01),黄芪汤高剂量组可见肾小球系膜细胞增生情况明显改善,肾小管轮廓清晰。结论: 黄芪汤对¹²C⁶⁺离子辐射脑模型鼠的肾损伤具有一定的防护作用,其作用机制可能与调控Bcl-2/NF-κB信号通路有关。     

Effects of Different Ambient Temperatures on Caecal Microbial Composition in Broilers

Short-term or acute temperature stress affect the immune responses and alters the gut microbiota of broilers, but the influences of long-term temperature stress on stress biomarkers and the intestinal microbiota remains largely unknown. Therefore, we examined the effect of three long-term ambient temperatures (high (HC), medium (MC), and low (LC) temperature groups) on the gene expression of broilers’ heat shock proteins (Hsps) and inflammation – related genes, as well as the caecal microbial composition. The results revealed that Hsp70 and Hsp90 levels in HC group significantly increased, and levels of Hsp70, Hsp90, IL-6, TNF-α, and NFKB1 in LC group were significantly higher than in MC group (p < 0.05). In comparison with the MC group, the proportion of Firmicutes increased in HC and LC groups, while that of Bacteroidetes decreased in LC group at phylum level (p < 0.05). At genus level, the proportion of Escherichia/Shigella, Phascolarctobacterium, Parabacteroides,and Enterococcus increased in HC group; the fraction of Faecalibacterium was higher in LC group; and the percentage of Barnesiella and Alistipes decreased in both HC and LC groups (p < 0.05). Functional analysis based on communities’ phylogenetic investigation revealed that the pathways involved in environmental information processing and metabolism were enriched in the HC group. Those involved in cellular processes and signaling, metabolism, and gene regulation were enriched in LC group. Hence, we conclude that the long-term temperature stress can greatly alter the intestinal microbial communities in broilers and may further affect the host’s immunity and health.     

Oxidation-induced calcium-dependent dehydration of normal human red blood cells

Phenazine-methosulphate (PMS) is a strong oxidant that induces reactive oxygen species (ROS) formation in cells. Though it has been shown that PMS increases the red blood cell (RBC) membrane permeability to K⁺, the hypotheses on the mechanism of PMS-induced effects are contradictory and there are no data on volume changes induced by this oxidant. Therefore, the influence of the PMS + ascorbate oxidative system on the volume of normal human RBCs was studied. In a Ca^{2 +} -containing medium, PMS + ascorbate caused dehydration (shrinking) of RBCs judged by: (1) changes in the density and osmotic resistance distributions of RBCs, and (2) a decrease in their low-angle scattering assessed by FACS analysis. The dehydration resulted from activation of the Gardos channels, was PMS and ascorbate concentration-dependent, was associated with broadening of the density and osmotic resistance distributions of the RBCs, and decreased in the presence of the taxifolin and rutin antioxidants. These findings contribute to a better understanding of the physiology and pathology of oxidatively-modified RBCs and may be of practical significance in estimating the antioxidant activity of various substances.     

口腔癌中肿瘤出芽与VEGF—C表达及淋巴结转移的相关关系

目的：通过检测肿瘤出芽、淋巴结转移以及血管内皮生长因子-C（VEGF—C）表达水平,分析口腔癌中肿瘤出芽与VEGF—C表达及淋巴结转移的相关关系,为临床治疗提供理论参考。方法：选取2009年1月-2013年1月4年间在我院接受诊治且资料完整63例口腔癌患者作为研究对象,观察肿瘤出芽、VEGF-C表达和淋巴结转移情况,分析相互之间的相关关系。结果：本次纳入研究的患者中,检出肿瘤出芽患者40例,所占比例为63．5％,VEGF—C表达阳性患者39例,阳性率率为61．9％,淋巴结转移患者40例,转移率为63．5％;肿瘤出芽与淋巴结转移的符合率为84．1％,肿瘤出芽与VEGF—C的表达符合率为79．4％,VEGF-C的表达与淋巴结转移发生的符合率为76．2％。肿瘤出芽与淋巴结转移呈正相关,经Spear相关分析,r=0．932,P〈0．05,与VEGF-C的表达也呈正相关,经Spear相关分析,r=0．897,P〈0．05。结论：肿瘤出芽与VEGF—C的表达水平和淋巴结转移均呈正相关关系,可用于预测判断口腔癌淋巴结转移情况。     

Chlorophyll fluorescence in the leaves of Tradescantia species of different ecological groups: Induction events at different intensities of actinic light

Chlorophyll fluorescence analysis is one of the most convenient and widespread techniques used to monitor photosynthesis performance in plants. In this work, after a brief overview of the mechanisms of regulation of photosynthetic electron transport and protection of photosynthetic apparatus against photodamage, we describe results of our study of the effects of actinic light intensity on photosynthetic performance in Tradescantia species of different ecological groups. Using the chlorophyll fluorescence as a probe of photosynthetic activity, we have found that the shade-tolerant species Tradescantia fluminensis shows a higher sensitivity to short-term illumination (≤20 min) with low and moderate light (≤200 μE m⁻² s⁻¹) as compared with the light-resistant species Tradescantia sillamontana. In T. fluminensis, non-photochemical quenching of chlorophyll fluorescence (NPQ) and photosystem II operational efficiency (parameter Φ_PSII) saturate as soon as actinic light reaches ≈200 μE m⁻² s⁻¹. Otherwise, T. sillamontana revealed a higher capacity for NPQ at strong light (≥800 μE m⁻² s⁻¹). The post-illumination adaptation of shade-tolerant plants occurs slower than in the light-resistant species. The data obtained are discussed in terms of reactivity of photosynthetic apparatus to short-term variations of the environment light.     

Ion channels of glutamate receptors: structural modeling

Ionotropic glutamate receptors belong to the superfamily of P-loop channels as well as K⁺, Na⁺, and Ca²⁺ channels. However, the structural similarity between ion channels of the glutamate receptors and K⁺ channels is a matter of discussion. The aim of this study was to analyze differences between the structures of K⁺ channels and glutamate receptor channels. For this purpose, homology models of NMDA and AMPA receptor channels (M2 and M3 segments) were built using X-ray structures of K⁺ channels as templates. The models were optimized and used to reproduce specific data on the structure of glutamate receptor channels. Particular attention was paid to the data of the binding of channel blockers and to the results of scanning mutagenesis. The modeling demonstrates that properties of glutamate receptor channel can be reproduced assuming only local structural deformations of the K⁺ channel templates. The most valuable differences were found in the selectivity-filter region, whereas helical parts of M2 and M3 segments could have similar spatial organization with homologous segments in K⁺ channels. It is concluded that the current experimental data on glutamate receptor channels does not reveal global structural differences with K⁺ channels.     

太子参红外指纹图谱的双指标序列分析

本文采用傅里叶变换红外光谱仪测定了四大种植主产区(安徽、江苏、福建、贵州)的栽培品及河南的野生品,共13个批次太子参甲醇提取物,获得不同产地太子参栽培品的FTIR指纹图谱,并对指纹图谱进行了相似度的计算,同时结合(共有峰率,变异峰率)双指标序列分析方法,对不同产地太子参的FTIR光谱进行了研究。结果表明,该方法所建立的太子参FTIR指纹图谱能够区分栽培品和野生品的差异,双指标序列分析法能够为产区间和产区内太子参品质差异提供参考,两种分析方法结合可为该药材的质量分析、评价与控制提供依据。     

单纯疱疹病毒1型引起的发热出疹性疾病需要与手足口病进行鉴别

对2008年在甘肃省发生的一起疑似手足口病(HFMD)的发热出疹性疾病的流行进行病原体的实验室检测,明确引起这起传染病流行的病原体。从4名发热出疹患者采集的8份临床标本中(每个患者采集咽拭子和疱疹液标本),首先将临床标本接种到RD和HEp-2细胞上进行病毒分离,对病毒分离阳性的标本提取病毒核酸,然后使用荧光定量-逆转录-聚合酶链式反应(RT-PCR)进行人肠道病毒(HEV)核酸的检测。对HEV检测结果阴性的标本采用序列非依赖的单引物扩增技术(SISPA)进行"未知病原体"的鉴定。分离到的6株病毒均鉴定为单纯疱疹病毒I型(HSV-1),结合分析这起流行中患者的临床表现、流行病学调查结果以及实验室检测结果,表明HSV-1是引起这起发热出疹性疾病流行的病原体。6株HSV-1的gG区核苷酸和氨基酸水平上差异都较小,同源性分别高达98.8%和97.9%,说明这起疫情是由同一个病毒传播链引起的。HSV-1等病毒引起的发热出疹性疾病需要与HFMD进行鉴别诊断,由于仅从临床症状和流行病学资料上判断引起发热出疹性疾病的病原体是非常困难的,因此,必须依赖于实验室的诊断。     

陕西省烟草灰霉病病原及云芝多糖对其防治研究

采用形态学观察和分子鉴定方法对2011年在陕西省发生的一种烟草未知病害的病原菌进行鉴定。从病叶组织分离纯化得到病原菌,通过致病性测定以及人工接种后再分离病菌,证明编号LJL007的菌株为该病的致病菌。依据病原菌的形态学和培养特征,将菌株LJL007鉴定为灰葡萄孢Botrytis cinerea Pers.,其有性型为富氏葡萄孢盘菌Botryotinia fuckeliana Whetzel。通过核糖体DNAITS序列分析,分离菌株LJL007序列(登录号:HM17900)与富氏葡萄孢盘菌序列(登录号:HM849615)同源性达100%,进一步证明该病原菌是灰葡萄孢Botrytis cinerea。云芝多糖在离体条件下,对灰葡萄孢的菌丝生长和孢子萌发均无直接抑制作用。云芝多糖对烟草灰霉病有较好预防保护作用,其预防效果可达56.29%。云芝多糖可显著提高烟草体内几丁质酶和β-1,3-葡聚糖酶活性,其活性峰值分别比对照提高56.89%和429.83%,说明云芝多糖可诱导植物产生抗病性。     

紫花苜蓿F-box蛋白基因MsFTL的克隆及功能分析

FTL(F-box Triple LRR protein)是F-box蛋白家族的成员,具有F-box保守结构域,在植物抵御逆境胁迫过程中起重要作用。本研究参考低温胁迫下紫花苜蓿转录组数据设计引物,通过RT-PCR克隆获得紫花苜蓿MsFTL基因,该基因的全长1422 bp,编码473个氨基酸。该蛋白含有1个F-box结构域及3个LRR重复。系统进化分析表明,MsFTL与蒺藜苜蓿XP_003626345.1 F-box/FBD/LRR-repeat protein亲缘关系最近。两者蛋白序列比对发现共有11个差异位点。在低温、盐、干旱以及外源ABA处理下,MsFTL基因受到诱导,表达量上调。构建植物过表达载体pCBM-MsFTL,通过农杆菌介导法转化烟草。对经过抗性筛选、PCR和Real-time PCR验证的转基因植株进行低温抗性鉴定。在-4℃低温胁迫下,野生型烟草叶片出现了明显的萎蔫失水现象,而转基因烟草萎蔫程度相对较轻。生理检测结果表明,4℃处理24 h之后,转基因烟草的可溶性蛋白含量、可溶性糖含量、SOD活性,CAT活性高于野生型,MDA含量低于野生型。本研究表明,MsFTL基因在提高植物对低温胁迫的抗性方面具有重要的作用。