丙型肝炎病毒准种多样性与病毒血症水平、疾病活动度及干扰素疗效的关系

目的　了解丙型肝炎病毒 (HCV)准种变异与病毒血症水平、疾病活动度及干扰素疗效的关系。方法　采用针对HCVE2高变区 1 (HVR1 )的单链构象多态性分析法 (SSCP)对 68例慢性丙型肝炎患者进行HCV准种检测 ,分析准种数目与HCVRNA、ALT、AST水平及肝组织活动指数 (HAI)的相关性。对其中 48例给予干扰素治疗 ,分析准种数目对干扰素应答效果的影响。结果　 61例HVR1SSCP阳性 ,HCV准种数目为 (6 2± 2 4)条。准种数量与HCVRNA水平显著相关 (P <0 0 1 ) ,与ALT、AST及HAI无明显相关 (P >0 0 5)。干扰素治疗患者中 ,43例HVR1阳性 ,持续应答者治疗前HCV准种数量 (3 3± 1 2 ,n =1 1 )显著少于获得治疗终点应答 (ETR)伴复发者 (6 3± 2 2 ,n =1 2 ,P <0 0 5)或无应答者 (8 0± 3 3 ,n=2 0 ,P <0 0 1 )。治疗结束时 ,干扰素组仍有 1 6例检测出HCV准种 ,但准种条数降为 (3 4± 1 2 )条 ,与未接受干扰素治疗病例的准种数目 (6 8± 2 5)相比差异有显著性 ,P <0 0 1 ) ,且其中 1 0例准种模式发生了改变。结论　HCV准种多样性可引起较高的病毒血症水平 ,但与疾病活动度无关 ;准种数目可作为预测慢性丙型肝炎干扰素疗效的指标。     

1b型丙型肝炎病毒基因组不同区域基因变异对干扰素疗效的影响

本研究对14例基因型1b型慢性丙型肝炎患者血清HCV基因组中E2/NS1(含HVR1和HVR2)、NS5A区准种异质性进行了检测,对HVR区准种异质性程度与干扰素(IFN)疗效的关系、NS5A2209-2248区是否存在干扰素敏感决定区(ISDR)等热点问题进行了探讨,为1b型慢性丙型肝炎患者应用IFN抗病毒治疗时     

慢性丙型肝炎患者血清细胞因子水平的变化及意义

目的:检测慢性丙型肝炎患者血清细胞因子IL-2、IFN-г、IL-5、IL-6、IL-12P70和P40水平,探讨其与患者血清ALT水平、HVC RNA载量、HCV基因型及干扰素疗效的关系。方法:检测30例健康对照者和30例慢性丙型肝炎患者干扰素治疗前后血清IL-2、IFN-г、IL-5、IL-6、IL-12P70和P40的含量,比较干扰素治疗应答组和无应答组之间细胞因子水平的差异及上述细胞因子水平与血清ALT水平、HCV基因型、HCVRNA载量等的关系。血清细胞因子检测应用ELISA法,HCV基因分型应用直接测序法,HCVRNA载量采用荧光定量PCR法。结果:与健康对照组相比,慢性丙型肝炎患者血清IL-2含量明显降低,IL-5和IL-12P40明显生高;血清IL-6含量与血清ALT水平呈正相关,与RNA载量呈负相关;HCV基因型1型患者血清IL-6含量明显高于2型,其他基因型和亚型之间细胞因子水平均无显著性差异;干扰素治疗的持续应答率为46.7%,应答组和无应答组治疗前血清细胞因子水平均无显著性差异,但应答组治疗结束时IFN-γ含量较治疗前明显升高。结论:血清Th1/Th2细胞因子水平失衡与丙型肝炎的慢性化和肝脏炎症活动相关;干扰素治疗前血清Th1/Th2细胞因子水平与干扰素治疗效果无关,不能对疗效进行预测,干扰素诱导的Th1细胞优势反应与持续应答有关。     

血清Th1/Th2细胞因子水平与慢性丙型肝炎临床及干扰素疗效关系的研究

目的 探讨血清Th1和Th2细胞因子水平与慢性丙型肝炎患者病情进展及干扰素疗效的关系.方法 血清细胞因子检测应用ELISA法,HCV基因分型应用直接测序法,HCVRNA载量采用荧光定量PCR法.结果 慢性丙型肝炎患者血清IL-2和TGF-β含量明显低于健康对照组,IL-5含量明显高于后者;血清IL-6含量与ALT水平呈正相关,与RNA载量呈负相关;HCV基因型1型患者IL-6含量明显高于2型,2a型IL-2含量低于2b型;随着感染时间的延长,血清IL-2呈下降趋势,IL-6呈升高趋势,而TGF-β则先升高之后逐渐下降.干扰素治疗应答组和无应答组治疗前血清细胞因子水平均差异无统计学意义,但应答组治疗结束时IFN-γ含量较治疗前明显升高.结论 慢性丙型肝炎患者存在Th1/Th2细胞因子失衡并与临床表现相关;治疗前血清Th1/Th2细胞因子水平不能对疗效进行预测,干扰素诱导的Th1细胞优势反应与持续应答有关.     

丙型肝炎病毒包膜2区准种复杂性及其临床意义初探

目的 研究丙型肝炎病毒（HCV）包膜2（E2）区准种复杂性及其临床意义。方法 以9例不同血清丙氨酸转氨酶水平物急性HCV感染者和4例慢性丙型肝炎患者为对象，通过逆转录PCR扩增HCVE2区氨基端片段（560bp)，扩增产物直接克隆，以单链构象多态性（SSCP）/异质性双体（HD）分析对每份血清标本的30个克隆进行筛选及准种复杂性分析，结果 慢性丙型肝炎患者血清HCV准种复杂性明显增高：HCV感染者血清ALT水平变化与准种复杂性无明显关系。结论 HCVE2区准种复杂性与病毒持续性感染有关。征收肝损害程度无关。     

聚乙二醇干扰素α-2a联合利巴韦林治疗高病毒载量1b基因型丙型肝炎患者无病毒学应答的预测因素分析

目的 探讨聚乙二醇干扰素α-2a联合利巴韦林治疗高病毒载量1b基因型HCV患者无病毒学应答的预测因素.方法 选取于2010年2月至2014年12月我院接受的聚乙二醇干扰素α-2a联合利巴韦林治疗的HCV基因1b型感染的慢性HCV初治患者72例.治疗3周后检测病毒学应答状态,无应答组患者23例,有应答组患者49例.回顾性统计患者的一般临床资料及相关指标,并做统计学分析.结果 有应答组患者HCV病毒RNA水平及HCV病毒核心抗原水平均明显下降,在第1天到第14天的每个时间点上两组比较差异均具有统计学意义(P ＜0.001).最高的HCV病毒RNA水平HCV病毒核心抗原水平曲线下面积均在第2周发生.第2周HCV病毒RNA消耗及耗损准确性均最高,其准确性与白细胞介素-28B minor+C区氨基酸70替代突变的准确性一致.结论 从治疗开始后2周内的病毒动力学分析,特别是IL-28B基因多态性和病毒突变,2周时的病毒应答情况是最有效的无病毒学应答的预测因素.因此,在2周时病毒下降有可能作为重要的预测指标.     

丙型肝炎病毒包膜区变异与感染慢性化关系的初步研究

目的 探讨丙型肝炎病毒(HCV)包膜区变异与其感染慢性化的关系。方法 3份HCV慢性感染者和3份急性感染者血标本，采用逆转录—聚合两链反应(RT—PCR)扩增HCV的E1区C端及E2区N端片段(573bp)，扩增产物进行克隆，以单链构象多态性(SSCP)和异质性双体(HD)分析对每份血清30个克隆的E1／E2区准种(quasispecies)进行筛选，挑选每份标本HCV的优势株与劣势株序列进行测定，分析非同义替换碱基数与同义替换碱基数比率(dN／dS，间接反映选择压力)和推导的氨基酸序列。结果 HCV慢性感染者病毒准种的复杂性和E2区dN／dS明显高于急性感染者。HCV慢性感染者的E2区氨基酸替换率(8．46％)比急性感染者(1．02％)更高，而两者的E1区氨基酸替换率(分别为2．74％和1．09％)均较低。尽管HVR1变异程度更高，但仍存在高度保守的氨基酸位点。结论 HCV持续性感染与准种复杂性增高和宿主对HVR1的免疫选择有关。     

67例慢性丙型肝炎治疗过程及随访中血清ALT、病毒载量及肝纤维化指标的变化

目的 观察聚乙二醇干扰素α-2b联合利巴韦林治疗慢性丙型肝炎过程中肝功能、病毒复制及肝纤维化指标的改变情况.方法 检测67例慢性丙型肝炎患者在干扰素联合利巴韦林治疗开始（0周）、结束（48周）和停药12周（60周）时血清丙氨酸转氨酶（ALT）、丙肝病毒核糖核酸（HCV RNA）、透明质酸（HA）、Ⅲ型前胶原肽（PCⅢ）、Ⅳ型胶原（Ⅳ-C）和层粘连蛋白（LN）水平.结果 治疗后完全应答组（CR-S,43/67） ALT、HCV RNA及血清4项纤维化指标均显著下降（P ＜0.05或P＜0.01）,部分应答组（CR-R,13/67）和无应答组（NR,11/67） ALT、HCV RNA及血清4项纤维化指标变化不明显,反跳甚至更高.结论 聚乙二醇干扰素α-2b联合利巴韦林治疗慢性丙型肝炎约65％患者完全应答,随着肝细胞炎症的改善,病毒RNA滴度、肝纤维化指标水平明显下降,表明干扰素联合利巴韦林能抑制HCV RNA复制,调节机体免疫功能,减轻肝脏炎症反应,改善肝功能,减少肝纤维化.     

慢性1b型丙型肝炎病毒感染者包膜2基因正选择位点分析及功能意义

目的:纵向分析自然免疫压力下慢性1b型HCV感染者E2基因的系统进化模式及正选择位点,初步探讨E2基因正选择位点的分布与已知功能区之间的关系。方法:未抗病毒治疗的3例慢性1b型HCV感染者及其系列血清样本均来自中国人HCV感染患者系列标本库(CQueen cohort)。对每例感染者3个连续时间点的血清样本进行HCV5′端6000bp基因组的扩增、克隆,每个时间点随机挑选25～33个克隆对E2基因测序,用MEGA软件构建系统进化树,用固定效应似然法检测HCV E2基因的正选择位点。结果:3例HCV慢性感染者的9份血清样本均获清晰、无杂带的6030bp的目的片段。慢性HCV感染者体内病毒准种的复杂程度并非随着感染时间的延长而进行性增加,在某个时间点上准种变异株的序列可能会趋于一致。准种优势变异株在间隔半年时间以上已发生更替。对每位患者动态的准种变异株进行正选择位点的分析,共检测到5个氨基酸(aa)位点:aa384、aa399、aa410、aa475和aa522,分布于HCV E2基因HVR1区、HVR2区及HCV E2-CD81分子结合区。结论:基于动态变化的HCV准种优势株的相关研究,可考虑选择间隔半年以上时间点的血清样本作为研究对象。慢性HCV感染过程中,HCV E2基因位于重要功能区的aa位点受到了宿主强烈的免疫压力,aa384、aa399及aa410位点的变异可能导致HCV发生体液免疫逃逸;而aa475和aa522位点的变异则可能通过影响HVR2区的功能或HCV E2-CD81分子的亲和力或改变CD81分子的结构而导致HCV发生先天免疫逃逸。该研究为分析HCV C-NS3基因片段的正选择位点奠定了良好的基础。     

HCV 1b亚型ISDR变异影响PEG-IFN/RBV抗病毒疗效的新特点分析

目的探讨HCV干扰素敏感决定区（ISDR）氨基酸（aa）序列变异度对聚乙二醇干扰素（PEG-IFN）联合利巴韦林（RBV）治疗1b亚型慢性丙型肝炎（CHC）疗效的影响。方法 58例HCV1b亚型慢性感染者采用PEG-IFN/RBV联合方案治疗48周,并随访24周。定量检测血清HCV RNA,逆转录-聚合酶链反应扩增治疗前血标本中HCV ISDR片段并测序,MEGA4分析氨基酸序列变异度;二分类Logistic回归分析各变量与持续病毒学应答（SVR）之间的关系。结果治疗前血清HCV的ISDR氨基酸序列与HCVJ株比较,15例为野生型（未突变）,42例为中间型（1-3个突变）,1例为突变型（≥4个突变）。其中2218位点突变最多,约为60.3%（35/58）。ISDRaa突变数目与SVR关系密切（P=0.000）,ISDRaa突变数≥2的CHC组所获得的EVR和SVR明显高于aa突变数〈2的CHC组（P=0.041/P=0.012）。结论华南HCV1b亚型ISDR突变型（氨基酸变异≥4）极少。ISDRaa变异能够预测SVR;采用PEG-IFN/RBV联合方案治疗,对SVR有预测价值的ISDRaa突变数可由4个减少为2个。     

Asymmetry of the internal connections of the striate cortex of the cat in the projection zone of the center of the field of vision

Studies were carried out on the organization of the internal connections of the striate cortex in cats in the projection zone of the center (0–5°) of the field of vision by microintophoretic application of horseradish peroxidase to electrophysiologically identified orientational columns. The area containing neurons showing retrograde labeling in most cases extended in the mediolateral direction. Labeled cells were located in the upper (II, III) and lower (V, VI) layers of the cortex, and the shapes and orientations of the areas containing labeled neurons in these layers coincided. Spatial asymmetry was detected in the distribution of labeled neurons relative to the orientational column studied. Labeled cells were located predominantly medial to the columns, regardless of the distance from the projection of the area centralis. Considering the visuotopical map of field 17, the asymmetry detected here provides evidence that neurons in orientational columns have more extensive connections with neurons of the peripheral part of the cortex. An asymmetrical distribution of “silent” zones around the receptive fields of neurons in orientational columns is suggested, and that these appear to receive influences from the periphery of the visual field. Laboratory of Visual Physiology and Laboratory of Central Nervous System Morphology, I. P. Pavlov Institute of Physiology, Russian Academy of Sciences, 6 Makarov Bank, 199034 St. Petersburg, Russia. Translated from Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 82, No. 12, pp. 23–29, December, 1996.     

Effects of the lesion of the postcommissural part of the septum on the behavior of the rat.

The effects of the lesion of the postcommissural part of the septum on behavior of the rat has been studied. Results may be summarized as follows. An increase in the exploratory behavior in the open field which decreases rapidly; a decrease in the number of defecations in this test and a decrease in time leaving a dark environment for exploration. In the shuttle box test, no facilitation of the acquisition, but a permanent and quite significant increase in the intertrial activity has been found. We conclude that the lesions tend to decrease the emotivity of the subjects. An interpretation on the basis of the species -- specific defensive reactions explains the transitory and permanent effects of the lesions on the spontaneous activity.     

The development of the ligament of the head of the femur

The hip joints of 30 human male and female fetuses and stillborns between 20 mm and 350 mm crown-rump length were studied by light microscopy. The ligament of the head of the femur developed in situ as a condensation of mesenchyme at the end of the second month of intra-uterine life (IUL), and was vascularized by branches of acetabular vessels early in the fourth month. In the majority of fetuses older than 5.5 months IUL, vessels in the ligament passed a short way into the femoral head within cartilage canals, to supply a small region around the fovea capitis. The remainder of the head was supplied by vessels in canals from around the upper part of the neck. The ligament changed from predominantly cellular to fibrous during the last 4 months of IUL. This increase in strength suggested significant mechanical functions in utero: limitation of adduction-flexion and opposition to postero-superior dislocation were the most likely.     

Counterpointing the functional role of the forebrain and of the brainstem in the control of the sleep-waking system

This paper reviews the lifetime contributions of the author to the field of sleep-wakefulness (S-W), reinterprets results of the early studies, and suggests new conclusions and perspectives. Long-term cats with mesencephalic transection show behavioral/polygraphic rapid eye movement sleep (REMS), including the typical oculo-pupillary behavior, even when the section is performed in kittens prior to S-W maturation. REMS can be induced as a reflex. Typical non-rapid eye movement S (NREMS) is absent and full W/arousal is present only after a precollicular section. The isolated forebrain (IF) rostral to the transection exhibits all features of W/arousal and NREMS [with electroencephalographic (EEG) spindles and delta waves], arousal to olfactory stimuli, and including the appropriate oculo-pupillary behaviors. These features also mature normally after neonatal transection. REMS is absent from the IF. After deprivation there is NREMS pressure and rebound in the IF, but the decerebrate cat only shows pressure for REMS. Most IF reactions to pharmacologic agents are within expectations, except for the tolerance/withdrawal effects of long-term morphine use which are absent. In contrast, these effects are supported by the brainstem (i.e. seen in the decerebrate cat). In cats with ablation of the telencephalon, or diencephalic cats, delta waves are absent in the thalamus. EEG thalamic spindle waves are seen triggering S for only 4-5 days after ablation. Therefore, true NREMS is absent in chronic diencephalic cats although pre- and postsomniac behaviors persist. These animals are hyperactive and show a pronounced, permanent insomnia; however, a low dose of barbiturate triggers a dramatic REMS/atypical NREMS rebound. Cats without the thalamus (athalamic cats), initially show a dissociation between behavioral hyperactivity/insomnia and the neocortical EEG, which for 15-20 days exhibits only delta and slower oscillations. Fast, low-voltage W rhythms appear later on, first during REMS, but spindle waves and S postures are absent from the start, such that these cats also display only atypical NREMS. Athalamic cats also show barbiturate-sensitive insomnia. Cats with ablation of the frontal cortices or the caudate nuclei remain permanently hyperactive. They also show a mild, but significant hyposomnia, which is permanent in afrontal cats, but lasts for about a month in acaudates. The polygraphic/behavioral features of their S-W states remain normal. We conclude and propose that: (a) the control of the S-W system is highly complex and distributed, but is organized hierarchically in a well-defined rostro-caudal manner; the rostral-most or highest level (telencephalon), is the most functionally complex/adaptative and regulates the lower levels; the diencephalic/basal forebrain, or middle level, has a pivotal role in inducing switching between S and W and in coordinating the lowest (brainstem) and highest levels; (b) W can occur independently in both the forebrain and brainstem, but true NREMS- and REMS-generating mechanisms exist exclusively in the forebrain and brainstem, respectively; (c) forebrain and brainstem S-W processes can operate independently from each other and are preprogrammed at birth; this helps understanding normal and abnormal polygraphic/behavioral dissociations in humans and normal dissociations/splitting in aquatic mammals; (d) NREMS homeostasis is present in the IF, but only REMS pressure after deprivation persists in the decerebrate cat; (e) the thalamus engages in both NREMS and W; (f) insomnia in diencephalic cats is the result of an imbalance between antagonistic W- and S-promoting cellular groups in the ventral brain (normally modulated by the telencephalon); (g) the EEG waves, which are signature for each S-W state, appear to truly drive the concomitant behaviors, e.g. a hypothetical human IF could alternate between behavioral NREMS and W/arousal/awareness; (h) a role for REMS is to keep the individual sleeping at the end of the self-limiting NREMS periods. The need for accelerating research on telencephaling NREMS periods. The need for accelerating research on telencephalic S-W processes and downstream control of the lower S-W system levels is emphasized.     

Topical organization of the projections of neurons of the substantia nigra and the ventral field of the tegmentum of the midbrain to the head of the caudate nucleus of the cat

Neuroscience and Behavioral Physiology -     

Involvement of the striatum in the central regulation of the hormonal functions of the gonads

Studies reported here show that intrastriatal administration of corticoliberin to rats decreases the blood testosterone level. However, in conditions of chemical deficiency of dopaminergic transmission in the dorsal striatum induced by injection of 6-hydroxydopamine, the effect of this neurohormone did not appear. It is concluded that extrahypothalamic corticoliberin is involved in regulating the hormonal reproductive system acting via dopaminergic mechanisms. Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 85, No. 4, pp. 594–597, April, 1999.     

Ultrastructure of the endothelium of the drainage system of the eye

The endothelium of the ocular drainage system (Schlemm’s canal, collector tubules, and aqueous veins) in primary juvenile glaucoma undergoes degenerative dystrophic changes with compensatory hypertrophy and proliferation at the initial stages of the glaucomatous process and atrophy and desquamation at advanced and terminal stages. Progressive decrease in the pinocytous function of endotheliocytes, reduction of the protein-synthesizing and mitochondrial compartments of the cytoplasm, and formation of autophagosomes reflect the process of endotheliocyte degeneration in general. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 5, pp. 574–577, May, 2008