山东省柯萨奇病毒B1型VP1区全基因序列分析

为了解柯萨奇病毒B1型(Coxsackievirus B1,CV-B1)山东地方株的分子流行病学特征,本研究对1994年至2015年山东省急性弛缓性麻痹(Acute flaccid paralysis,AFP)监测系统、环境污水监测和无菌性脑膜炎病例标本中分离到的CV-B1病毒进行了VP1序列测定、系统发生学分析和同源性分析。共分离到CV-B1病毒53株,其中AFP监测、污水和及脑炎标本各分离到41株、4株和8株。基于VP1完整编码区序列的系统发生学分析显示CVB1山东株与国内其他分离株属于一个大的分支,该分支内无国外分离株,国外分离株构成了其他两个分支。山东株之间的VP1核苷酸同源性为84.4%~100.0%,与其他国家分离株的同源性为77.9%~85.0%。研究结果表明,中国CV-B1分离株与国外株相比有较大的遗传差异,需要加强相关手足口病病毒学监测,关注不同传播链的新的基因亚型的肠道病毒的输入。     

天津市致病毒性脑炎柯萨奇病毒B组5型全基因组序列分析

肠道病毒是我国病毒性脑炎(Viral encephalitis,VE)的主要病原体。本文研究对4株引起VE的天津柯萨奇病毒B组5型(Coxsackievirus B5,CV-B5)分离株进行Illumina MiniSeq高通量测序,并对其全基因组特征、进化及重组特点进行分析。结果提示,4株CV-B5天津分离株的全基因组核苷酸和氨基酸序列同源性分别为84.5%~100.0%和98.1%~100.0%,与国内流行株的全基因组核苷酸序列同源性为83.2%~96.5%,氨基酸序列同源性为96.4%~99.4%。基于全基因组的系统进化分析将CV-B5流行株分为A-D四个基因型,其中天津与国内流行株均属于C基因型。C基因型进一步分为3个进化分支,而天津分离株处在两个不同的分支上。基于基因组各区段序列的系统进化与SimPlot重组分析结果显示,天津分离株15-39N、15-41N与埃可病毒30型(Echovirus 30,E-30)原型株在P3区3B、3C、3D区域均检测到重组信号。本研究有助于了解CV-B5的全基因组特点和重组规律,为相关疾病的防控提供依据。     

云南省2006～2010年非脊灰肠道病毒分子流行病学特征分析

为了解云南省非脊髓灰质炎(脊灰)肠道病毒(NPEV)的基因型分布及分子进化特征,对2006～2010年间从急性迟缓性麻痹(AFP)病例中分离到的105株NPEVs进行VP1区部分核苷酸扩增和序列测定。所获得的云南地方株基因序列与各基因型原型株进行核苷酸与氨基酸同源性比较,并与GenBank中选取的代表株构建基因进化关系树。结果分析显示:105株NPEVs分别属于HEV-A、HEV-B、HEV-C,其中HEV-A 18株(7个血清型)所占比例为17.1%;HEV-B 77株(22个血清型)所占比例为73.3%,表明云南省AFP病例中流行的NPEV还是以HEV-B为主;HEV-C 10株(4个血清型)所占比例为9.5%;没有分离到HEV-D组肠道病毒;基因进化树中各种血清型病毒与对应原型株及代表株聚集一起,除CA2、EV90和EV76外,云南地方株与原型株位于不同分支。相同型别的毒株在5年的流行过程中变异程度亦不同,亲缘关系远近不一,表明这些病毒在云南省存在不同的传播链。     

急性弛缓性麻痹病例中ECHO11病毒的基因特征分析

该文首次分析了我国ECHO11病毒的分子流行病学资料。1999~2004年,ECHO11病毒是山东省急性弛缓性麻痹(AFP)病例中分离到的优势毒株,2003年从山东省482例AFP病例中共分离到11株ECHO11病毒,其中相关的10例病例分布跨越山东省大部分地区,但发病日期集中在7月和12月。该研究试图通过对VP1编码基因全序列的测定和分析,为探讨ECHO11病毒与AFP之间的病因关系提供线索。分子流行病学研究提示,11株E-CHO11毒株都位于同一传播链,核苷酸同源性为97.2%~100%,氨基酸同源性为99.6%~100%,其中7月和12月的分离株之间相差8~9个核苷酸,氨基酸序列一致。这说明山东省2003年7月和12月分别发生了ECHO11病毒流行,但这些毒株与AFP的病因关系尚需进一步研究。11株病毒组成了A基因型中的一个新亚型,在进化树上单独呈密切相关的一簇,与同基因型内的其它亚型的核苷酸同源性为82.2%~84.7%,氨基酸同源性为94.8%~97.6%。     

2018年河南省肠道病毒环境监测结果分析

通过对2018年河南省环境污水中肠道病毒(Enterovirus,EV)的持续监测,了解河南省环境污水中脊灰病毒(PV)和非脊灰肠道病毒(NPEV)的血清型分布及流行情况.选取了河南省东、西、南、北、中部的五个城市,对每个城市有10万～30万人共同使用的下水管网的地区进行污水采样,每两个月采集1次,每次采集两份污水样品,采集持续1年,对污水样品进行浓缩,将浓缩液接种至RD细胞、L20B细胞和HEp-2细胞进行病毒分离.对病毒分离物进行VP1区核苷酸序列测定分析,采用最大似然法构建系统进化树对EV进行分子定型,并进行核苷酸相似性分析.共采集污水样品60份,分离到EV阳性毒株16株(26.67％),分别为8株CVB5、3株E7、3株E11、1株CVB5+E11混合株;1株PVIII;环境监测标本与急性弛缓性麻痹(Acute flaccid paralysis,AFP)病例标本,在济源市和周口市分离到的CVB5比例有一致性,郑州市和济源市分离到的E7比例有一致性;污水标本中阳性毒株的检出时间要早于AFP病例标本中同种毒株型别的检出时间1～3个月.来源于AFP病例监测和环境监测的CVB5之间的核苷酸相似性为94.7％～99.6％,来源于AFP病例监测和环境监测的E7之间的核苷酸相似性为92.5％～100.0％.来源于AFP病例监测和环境监测的CVB5属于同一个基因型,E7也是同一个基因型,甚至是同一个病毒传播链.河南省2018年建立了环境监测方法,是AFP病例监测的有益的补充,也可对人类肠道病毒病流行或暴发进行预测和预警.     

辽宁省首株柯萨奇病毒B组5型全基因组序列分析

研究引起辽宁地区手足口病的柯萨奇病毒B组5型（coxsackievirus B5,CV-B5）基因组特征。对2018年从辽宁省688份肠道病毒核酸阳性的标本中分离到的1株CV-B5进行高通量测序，并对其全基因组进行遗传进化分析。结果表明，CV-B5辽宁分离株与国内流行株的全基因组核苷酸序列同源性为78.5%～97%，氨基酸序列同源性为75.3%～96.7%。基于全基因组的进化分析将CV-B5流行株分为A～D四个基因型，辽宁分离株属于D基因型。通过重组分析发现其在P3区的3D区段发生重组。首次在辽宁地区手足口病患儿中分离出CV-B5，辽宁省分离株（LN2018-23-21/CHN/2018）可能为重组株。     

基于环境污水监测的埃可病毒11型的循环动态变化及其基因特征分析

为探讨山东省环境污水监测中埃可病毒11型(Echovirus 11,Echo11)的动态变化及分子流行病学特征,本研究对山东省2011~2012年环境污水监测分离到的Echo11毒株进行了VP1区核酸扩增和序列测定,并与1994~2010年山东省急性弛缓性麻痹(Acute flaccid paralysis,AFP)监测系统中Echo11分离株以及国外同型毒株进行同源性分析和系统发生学分析。结果显示:2011~2012年山东省济南市和临沂市共分离到Echo11 94株,其时间分布具有季节性特征,夏秋季高峰明显。分离毒株之间核苷酸和氨基酸同源性分别为89.5%~100.0%和95.4%~100.0%;与原型株(Gregory)之间核苷酸和氨基酸同源性分别为76.6%~79.7%和90.4%~92.5%。山东环境分离株全部属于A基因型,毒株之间核苷酸变异较大,提示山东地区存在多个传播链共循环。本研究描述了山东省环境污水中Echo11分离株的动态变化和遗传特征,结果证明环境污水监测是探索人类肠道病毒动态流行和遗传变异的重要手段。     

基于环境污水监测的埃可病毒11型的循环动态变化及其基因特征分析北大核心CSCD

为探讨山东省环境污水监测中埃可病毒11型（Echovirus 11,Echo11）的动态变化及分子流行病学特征,本研究对山东省2011~2012年环境污水监测分离到的Echo11毒株进行了VP1区核酸扩增和序列测定,并与1994~2010年山东省急性弛缓性麻痹（Acute flaccid paralysis,AFP）监测系统中Echo11分离株以及国外同型毒株进行同源性分析和系统发生学分析。结果显示：2011~2012年山东省济南市和临沂市共分离到Echo11 94株,其时间分布具有季节性特征,夏秋季高峰明显。分离毒株之间核苷酸和氨基酸同源性分别为89.5%~100.0%和95.4%~100.0%;与原型株（Gregory）之间核苷酸和氨基酸同源性分别为76.6%~79.7%和90.4%~92.5%。山东环境分离株全部属于A基因型,毒株之间核苷酸变异较大,提示山东地区存在多个传播链共循环。本研究描述了山东省环境污水中Echo11分离株的动态变化和遗传特征,结果证明环境污水监测是探索人类肠道病毒动态流行和遗传变异的重要手段。     

肠道病毒B组山东地方株的基因型分布

为明确源自急性弛缓性麻痹(AFP)病例的HEV-B组病毒山东地方株的基因型分布,探讨其优势基因型的变迁与疾病暴发之间的关系,本研究对山东省1994年~2008年AFP监测系统分离到的HEV-B组病毒进行了VP1区核酸扩增和序列测定。序列测定结果显示HEV-B山东地方株共包括29种基因型,其中CVA 1种(CVA9),CVB 5种(CVB1~5),ECHO 20种以及新型肠道病毒EV73、75、97。其中ECHO11、CVB3、ECHO6、ECHO14、ECHO25是AFP监测系统中最常分离到的B组病毒。同源性比较显示,相同血清型HEV-B山东地方株型内核苷酸同源性最小75.4%,最大99.6%,与原型株核苷酸同源性最小73.8%,最大85.2%,但氨基酸变异不大。研究表明,不同基因型病毒具有不同的时间循环模式,相同基因型毒株内部根据其遗传距离的远近又可划分为不同的基因亚型,从而帮助确定HEV的传播途径和传播范围。     

Changes in lactate dehydrogenase activity in chicken tissues in hyperthyreosis in ontogenesis

The lactate dehydrogenase activity in reactions of lactate oxidation and synthesis was studied in subfractions of the chicken brain, heart and liver at the embryonal, early postembryonal and adult stages of development after thyroxine administration. It has been shown that during embryogenesis thyroxine predominantly enhanced the rate of lactate oxidation in the mitochondrial tissues. A marked increase in the lactate synthesis was found in cytoplasm of the adult chicken tissues. Specificity of enzyme activity alterations was detected in the chicken brain during ontogenesis after thyroxine administration.     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

Scurvy in capybaras bred in captivity in Argentine

In order to determine if the absence of vitamin C in the diet of capybaras (Hydrochoerus hydrochaeris) causes scurvy, a group of seven young individuals were fed food pellets without ascorbic acid, while another group of eight individuals received the same food with 1 g of ascorbic acid per animal per day. Animals in the first group developed signs of scurvy-like gingivitis, breaking of the incisors and death of one animal. Clinical signs appeared between 25 and 104 days from the beginning of the trial in all individuals. Growth rates of individuals deprived of vitamin C was considerably less than those observed in the control group. Deficiency of ascorbic acid had a severe effect on reproduction of another population of captive capybaras. We found that the decrease in ascorbic acid content in the diet affected pregnancy, especially during the first stages. The results obtained suggest that it is necessary to supply a suitable quantity of vitamin C in the diet of this species in captivity.     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.