丙泊酚麻醉对老龄大鼠认知功能和海马神经元γ-氨基丁酸A受体表达的影响

目的观察丙泊酚麻醉对老龄大鼠认知功能及海马神经元γ-氨基丁酸A(GABA)受体表达的影响。方法50只SD老年大鼠随机分为丙泊酚组和对照组,每组25只。丙泊酚组大鼠腹腔注射1%丙泊酚中/长链脂肪乳注射液6 mL/kg,对照组腹腔注射等体积的生理盐水。麻醉后1 d进行Morris水迷宫实验,分别采用HE染色和尼氏体染色观察海马区神经细胞及尼氏体(Nissl体)形态学变化,Western Blot检测GABA蛋白量表达。结果麻醉后,2组大鼠肛温、心率、呼吸频率、血氧饱和度比较,差异均无统计学意义(P>0.05)。随着实验时间的延长,2组大鼠逃逸潜伏期、总里程数逐渐减小,丙泊酚组大鼠各时间点逃逸潜伏期、总里程均高于对照组,差异有统计学意义(P<0.05)。丙泊酚组大鼠穿越平台区域次数、时间小于、短于对照组,差异有统计学意义(P<0.05)。对照组海马区Nissl体存在于细胞浆及树突,染色较深,神经细胞排列整齐,形态规则;丙泊酚组Nissl体消失,神经细胞数量减少,细胞核破裂、丢失。丙泊酚组大鼠海马GABA蛋白表达量低于对照组,差异有统计学意义(P<0.05)。结论丙泊酚对大鼠认知功能有影响,并与海马区GABA的表达相关。     

40m 以浅潜艇艇员饱和暴露后脱险减压方法的研究

1991～1992年进行了两次8人次模拟20、40m 饱和暴露后脱险减压方法的人体实验研究。在20m 空气饱和暴露后直接出水.在水面进行压力转移,6人次中出现皮肤瘙痒和关节疼痛4人次,当再加压回到20m后,症状消失:水面间隔总时间为9～15min。在4人次j40m 空气饱和暴露后直接减压到22m(再进行水面转移),未发现任何减压病症状。实验期间还观察了:在高压下人体自身的实际耗氧及高分压氧对肺部的影响。实验结果为今后现场实际应用的饱和脱险减压方法、减压程序提供了依据。     

FasL的表达在结直肠癌免疫逃逸中的意义

目的研究结直肠癌中Fas配体（FasL）的表达及其在结直肠癌免疫逃逸中的意义。方法采用免疫组织化学染色法，检测80例结直肠癌组织中FasL表达及肿瘤浸润淋巴细胞（TIL）的数量。应用原位杂交法，检测80例结直肠癌组织连续切片的FasL的。RNA的表达。采用脱氧核糖核酸末端转移酶介导的缺口末端标记技术（TUNEL），对80例结直肠癌组织中凋亡的TIL及肿瘤细胞进行观察。结果80例结直肠癌组织FasL表达程度不等，不论是在同一组织切片不同部位或两组织切片间相比，FasL表达程度和范围都不均匀。FasL的mRNA的表达部位与FasL蛋白的表达部位相对应。FasL表达程度高的组织的TIL计数低于FasL表达低的组织（P〈0．05），同时其TIL凋亡指数高于FasL表达低的组织，而结直肠癌细胞的凋亡指数低于FasL表达程度低的组织（P〈0．01），TIL凋亡指数与胃癌细胞的凋亡指数呈负相关（r=-0．631，P〈0．01）。结论全占直肠癌细胞可通过表达FasL，诱导TIL发生凋亡，反击机体免疫系统，这可能是结直肠癌免疫逃避的重要机制之一。     

Dynamics of HIV replication in lymphocytes and the efficacy of protease inhibitors

Using a system that allows transfection of resting peripheral blood lymphocytes (PBLs) two questions were addressed: the kinetics of HIV replication from the state of proviral latency, and the impact of different parameters on the efficacy of protease inhibitors to control HIV replication. PBLs were transfected with an infectious full length HIV-DNA harboring a luciferase reporter gene and activated thereafter. Ritonavir was added at different times at doses ranging from to 0.06 to 1 microM. Viral expression was assessed by quantifying luciferase activity in cell extracts and levels of p24 HIV antigen in culture supernatants. After transfection and cell activation, intracellular expression of HIV proteins, as assessed by luciferase detection, occurred within 2 hr. HIV-gag p24 antigen was detected in culture supernatants between 6 and 8 hr post-activation. Ritonavir was effective in blocking viral replication when given within 4 hr following HIV reactivation, but a delay in ritonavir administration or breaches in ritonavir levels after 6 hr from transfection resulted in viral escape. HIV reactivation from proviral latency in PBLs is an extremely rapid process, faster than estimated from previous models. These data stress the need for maintaining effective antiretroviral concentrations to block completely viral replication.     

Epitope specificity is critical for high and moderate avidity cytotoxic T lymphocytes associated with control of viral load and clinical disease in horses with equine infectious anemia virus

Equine infectious anemia virus (EIAV) is a lentivirus that causes persistent infections in horses. We hypothesized that high-avidity CTL specific for nonvariable epitopes might be associated with low viral load and minimal disease in EIAV-infected horses. To test this hypothesis, memory CTL (CTLm) responses were analyzed in two infected horses with high plasma viral loads and recurrent disease (progressors), and in two infected horses with low-to-undetectable viral loads and mild disease (nonprogressors). High-avidity CTLm in one progressor recognized an envelope gp90 epitope, and the data documented for the first time in EIAV that viral variation led to CTL escape. Each of the nonprogressors had high-to-moderate avidity CTLm directed against epitopes within Rev, including the nuclear export and nuclear localization domains. These results suggested that the epitope specificity of high- and moderate-avidity CTLm was an important determinant for disease outcome in the EIAV-infected horses examined.     

肿瘤免疫编辑研究进展

在肿瘤发生发展的过程中,机体免疫系统与肿瘤细胞相互作用,相互编辑.免疫系统的监视作用可清除肿瘤或抑制肿瘤的发生发展,同时免疫系统又可对肿瘤细胞进行重塑,使肿瘤发生免疫逃逸.而肿瘤可诱发机体抗肿瘤的固有及获得性免疫应答,同时也可改变免疫系统的抗肿瘤效应.了解肿瘤免疫编辑的相关研究进展可以为肿瘤防治提供参考.     

Escape efficiency of diploid and polyploid frogs: A comparison ofOdontophrynus cultripes (2n=22) andO. americanus (4n=44)

The escape efficiency of two closely related species of frogs,Odontophrynus cultripes(2n=22) and the tetraploidO. americanus (4n=44), were compared in a shuttle box and under simulated naturalistic conditions.O. americanus was generally superior toO. cultripes, and females tended to outperform males within both species. The relative inefficiency ofO. cultripes escape behavior was examined in light of the animals' having an elaborate, passive defense mechanism in the form of well-marked venom glands. Escape efficiency was highly variable in both species. Possessing twice the amount of DNA, the tetraploid behavioral variation was paradoxically less than that of the diploid, but compatible with what has been found for morphological characters in other organisms.This research was carried out at the Instituto Butantan with the support of ongoing grants from the Brazilian CNPq, FAPESP, FEDIB, and PNUD while the first author was a visiting professor in the Departamento de Genética Humana, Instituto de Biociências, Universidade de São Paulo, under the auspices of the Programa Multinacional de Genética, Organization of American States.     

Sequential change of the hypervariable region of the hepatitis C virus genome in acute infection

Hepatitis C virus (HCV) infection is characterized by persistence of liver inflammation that often leads to end-stage liver disease, although the mechanisms are not fully understood. A hyper-variable region (HVR) has been reported in the E2/NS1 region of the HCV genome, in which striking diversity is found among different HCV isolates. To investigate the association of the HVR alterations with the clinical courses of HCV infection, a longitudinal analysis of the HVR in patients with acute HCV infection was carried out. Plasma samples were obtained at several times in three patients with acute hepatitis C. Plasma RNA was extracted and reverse transcribed, and DNA fragments that included the HVR were amplified by PCR. The sequences of the HVR were directly determined from the PCR products by the dideoxy chain termination method, from which amino acid sequences were deduced. In all cases, plasma HCV-RNA disappeared with the improvement of the initial alanine aminotransferase (ALT) elevation, but HCV-RNA reappeared about 1 year later with or without deterioration of the hepatitis. In a case of sporadic acute hepatitis, the HCV in the recurrent phase had seven amino acid substitutions in the HVR compared with that in the acute phase, although no amino acid changes were noted during the initial acute phase. In a case of posttransfusion hepatitis, a marked difference was observed between the acute and the recurrent phases, with an amino acid homology of 30% (8/27). The mutation rate of the HVR had a tendency to accelerate as the HCV infection progressed to the chronic stage. In conclusion, the HVR changes serially during the course of acute HCV infection, and these HVR changes may play a part in the chronicity of HCV infection. © 1994 Wiiey-Liss, Inc.     

Matrix protein mediated shutdown of host cell metabolism limits vesicular stomatitis virus-induced interferon-alpha responses to plasmacytoid dendritic cells

Upon infection with many different viruses, plasmacytoid dendritic cells (pDC) produce large amounts of type I interferon (IFN-/β). To address why upon vesicular stomatitis virus (VSV) infection pDC, but not conventional myeloid DC (mDC), are induced to produce IFN-, pDC and mDC were differentiated from bone marrow cells (BM-DC). Upon VSV infection BM-pDC produced IFN-, whereas BM-mDC did not. Notably, upon infection with VSV-M2, a VSV variant expressing a M51R mutant matrix (M) protein that showed a reduced sequestration of host cell metabolism, BM-pDC and BM-mDC mounted massive IFN- responses. Both DC subsets showed comparable RNA levels of retinoic acid inducible gene-I (RIG-I) and Toll-like receptor (TLR) 7 and were able to respond upon triggering with double-stranded RNA (dsRNA) or single-stranded RNA (ssRNA) analogs. Moreover, upon VSV-M2 infection IFN- production by both DC subsets was largely dependent on viral replication. Interestingly, upon virus infection BM-pDC, but not BM-mDC, up-regulated mRNA levels of nuclear export factors Nup96/98, probably reflecting cellular mechanisms to circumvent viral escape strategies. Collectively, these results indicated that cell types induced to produce IFN- upon viral infection are not primarily defined by cellular receptor configurations but rather by complex virus/host cell interactions.