含HPV16 E7的嵌合型VLPs引发细胞溶解反应

目的 探讨ＢＰＶ（牛乳头状瘤病毒）Ｌ１／ＨＰＶ（人乳头状瘤病毒）１６Ｅ７嵌合型乳头状瘤病毒样颗粒（ＶＩＰｓＸ）的免疫学特性。方法 将表达纯化的含ＢＰＶＬ１／ＨＰＶ１６Ｅ７的嵌合型ＶＬＰｓ作为抗原在体外刺激ＥＬ－４细胞并对其进行细胞毒性Ｔ淋巴细胞（ＣＴＬ）反应分析。结果 嵌合型ＶＬＰｓ可引发特异的ＣＴＬ反应。Ｌ１－ＶＬＰｓ抗体能阻断这种特异性细胞溶解反应。结论 含ＢＰＶＬ１／ＨＰＶ１６Ｅ７嵌合型的Ｖ     

肺淋巴上皮瘤样癌与EB病毒的相关性

目的为阐明肺淋巴上皮瘤样癌（ｌｙｍｐｈｏｅｐｉｔｈｅｌｉｏｍａ－ｌｉｋｅｃａｒｃｉｎｏｍａ，ＬＥＬＣ）与ＥＢ病毒（ＥＢＶ）的关系。方法对３０例肺ＬＥＬＣ和１９例肺非ＬＥＬＣ的石蜡包埋组织，采用原位杂交、免疫组化技术检测ＥＢＶ的存在情况及其基因表达特征。结果肺ＬＥＬＣ中原位杂交ＥＢＶ小ＲＮＡ（ＥＢＥＲｓ）阳性率为９３．３％（２８／３０）；而免疫组化潜在膜蛋白（ｌａｔｅｎｔｍｅｍｂｒａｎｅｐｒｏｔｅｉｎ，ＬＭＰ１）阳性率分别为５３．３％（１６／３０）和２３．３％（７／３０）；１９例病毒衣壳抗原（ｖｉｒｕｓｃａｐｓｉｄａｎｔｉｇｅｎ，ＶＣＡ）阳性率为２３．３％（７／３０）；１９例非ＬＥＬＣ肺癌中原位杂交检测ＥＢＥＲｓ、免疫组化检测ＬＭＰ１均为阴性。结论肺ＬＥＬＣ与ＥＢＶ密切相关，ＥＢＶ可能在肺ＬＥＬＣ发生发展中起重要的作用；ＥＢＶ在肺ＬＥＬＣ中主要以潜伏状态存在，只有少量病毒进入溶解周期。     

CD28与B7.1共刺激PBLs受肝癌细胞诱导后的TCR Vβ基因表达

目的 研究Ｔ细胞受体（ＴＣＲ）Ｖβ在外周血淋巴细胞（ＰＢＬｓ）中的表达。方法 用单克隆抗体ＣＤ２８＋Ｂ７．１（ＣＤ８０）共刺激正常人外周血淋巴细胞后作用于肝癌细胞（ＢＥＬ－７４０２）。结果 发现ＰＢＬＳＴＣＲＶβ７基因在体外选择性扩增,表达水平为１５￣２５％,对照组为４￣８％。结论Ｖβ７基因对识别肿瘤抗原具有特异性,为肿瘤基因治疗和研究提供新的线索和实验依据。     

无镁液灌流大鼠海马脑片自发性癫痫样放电的药物敏感性

用常规海马脑片实验方法：撤除人工脑脊液中的Ｍｇ＾２＋、０．５ｈ内ＣＡ１区出现自发性痫样放电,１ｈ内于稳定,约１／３脑片波幅可稳定２ｈ左右,但放电频率呈缓慢增加,无Ｍｇ＾２＋灌流后１￣３ｈ间可以观察抗痫药的作用。苯妥英钠（ＰＨＴ）１００μｍｏｌ／Ｌ,氨甲酰氮卓（ＣＢＺ）１００μｍｏｌ／Ｌ,安定（ＤＺＰ）３０μｍｏｌ／Ｌ,苯巴双妥钠（ＰＢ）１００μｍｏｌ／Ｌ,丙戊酸钠（ＶＰＡ）１０ｍｍｏｌ／Ｌ及乙琥胺     

EB病毒编码的RNA及EB病毒潜在膜蛋白在中线T淋巴瘤中的表达

应用免疫组化和原位杂交技术及抗ＥＢ病毒潜在膜蛋白（ＬＭＰ－１）单克隆抗体和ＥＢ病毒编码的ＲＮＡ（ＥＢＥＲ－１）探针对９例中线Ｔ淋巴瘤（ＭＴＬ）进行了ＥＢ病毒（ＥＢＶ）检测。结果显示：８例肿瘤细胞核ＥＢＥＲ－１阳性；７例肿瘤细胞膜和胞浆ＬＭＰ－１阳性。结果表明：（１）ＥＢＶ与我国的ＭＴＬ存在密切关系，很可能在其发病中起着重要作用；（２）ＥＢＶ在ＭＴＬ的检出率高于全身其它部位和其它类型的周围型Ｔ淋巴瘤；（３）ＥＢＥＲ－１原位杂交和ＬＭＰ－１免疫组化在检测ＭＴＬ中ＥＢＶ方面都很敏感、可靠，而后者更经济简便。     

EBV转化的人B淋巴母细胞和人B淋巴细胞杂交瘤细胞表面标记表达的研究

本文用碱性磷酸酶－抗碱性磷酸酶（ＡＰＡＡＰ）免疫组化染色技术，对ＥＢ病毒（ＥＢＶ）转化的人Ｂ淋巴母细胞（ＬＣＬ）、人Ｂ淋巴细胞杂交瘤以及它们的亲本骨髓瘤细胞系的部分表面标记进行了检测。结果显示，人Ｂ淋巴细胞在ＥＢＶ转化或杂交过程中表面标记发生了较大的变化；对ＬＣＬ和杂交瘤细胞表面标记的检测可能为探寻人Ｂ细胞在这些过程中染色体保留或丢失的规律提供一种手段。此外，ＬＣＬ及人Ｂ细胞杂交瘤细胞系有可能被用     

喉癌发生中人乳头瘤病毒感染与Langerhans细胞,p53表…

目的；探讨喉癌的发生机理。方法：应用免疫组化ＬＳＡＢ法对３０例喉鳞状细胞癌人乳头瘤病毒（ＨＰＶ）感染、Ｌａｎｇｅｒｈａｎｓ细胞（ＬＣ）及Ｐ５３慢白表达进行了研究。结果：２６．７％的病例可以检测到ＨＰＶ抗原成分。ＨＰＶ感染的癌旁粘膜内ＬＣ数量明显少于无感染者，且形态也发生改变。Ｐ５３蛋白表达阳性率在ＨＰＶ感染组明显低于ＨＰＶ检测阴性组。结论：提示ＨＰＶ、ＬＣ、Ｐ５３在喉癌发生发展过程中起一定作用，且     

用肾综合征出血热病毒结构蛋白体外免疫正常人外周血淋巴细胞制备单克隆抗体

用亲和层析提取的肾综合征出血热病毒（ＨＦＲＳＶ）结构蛋白（５０ｋＤ和６７ｋＤ）为抗原，在体外免疫正常人外周血淋巴细胞（ＰＢＬ）。ＰＢＬ经亮氨酸甲基酯处理后，在含有ｓＰＷＭ－Ｔ、ＩＬ－２及不同浓度抗原的培养基中培养６ｄ，计数存活ＰＢＬ数量。结果表明，在上述体外免疫条件下存活ＰＢＬ的数量与抗原剂量呈正相关。将此体外免疫的ＰＢＬ与人－鼠杂交瘤细胞（Ｋ６Ｈ６／Ｂ５）融合，获得了３株分泌抗ＨＦＲＳＶ人单抗的杂交瘤细胞，表明在体外免疫条件下，正常人ＰＢＬ能够对ＨＦＲＳＶ结构蛋白发生特异性免疫应答。     

系统性红斑狼疮患者B淋巴细胞EBV-LMP1和ZEBRA的表达研究

目的：探讨ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ在系统性红斑狼疮患者（ＳＬＥ）的表达情况。方法：间接荧光免疫标记，流式细胞仪检测。结果：ＳＬＥ患者Ｂ淋巴细胞中ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ的表达显著高于正常对照组（Ｐ＜０．０１）。活动期患者ＣＤ２３＋细胞ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ的表达率均高于ＣＤ１９＋细胞（Ｐ＜０．０１）。非活动期患者ＣＤ２３＋细胞ＥＢＶ－ＬＭＰ１表达也高于ＣＤ１９＋细胞（Ｐ＜０．０１）。但ＥＢＶ－ＺＥＢＲＡ表达在两亚群间差异没有统计学意义（Ｐ＞０．０５）。结论：朋病毒参与了ＳＬＥ的发病机制，病毒主要以潜伏期状态存在于患者中，病毒复制促进病情发展，检测Ｂ淋巴细胞ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ的表达率，有助于病情活动指标的判断。     

蓝舌病毒L3基因的克隆与表达

目的　通过高效表达,研究蓝舌病毒（ＢＴＶ） ＶＰ３ 的功能,为后续ＢＴＶ病毒样颗粒的装配作准备。方法　克隆出完整的ＢＴＶ１３ Ｌ３ 基因,将其插入杆状病毒表达载体进行表达。结果　获得了含有全长Ｌ３ 基因的克隆,ＶＰ３ 在昆虫细胞中得到了高效表达,表达蛋白占细胞总蛋白的１０％ ～１５％ ,ＶＰ３ 与ＶＰ７ 共表达可装配出ＢＴＶ 核心样颗粒。结论　在昆虫细胞中表达ＢＴＶＶＰ３ 蛋白具有生物学活性,可用于ＢＴＶ病毒样颗粒的装配研究     

Hemodynamic alterations in cirrhosis and portal hypertension

Portal hypertension (PHT) is associated with hemodynamic changes in intrahepatic, systemic, and portosystemic collateral circulation. Increased intrahepatic resistance and hyperdynamic circulatory alterations with expansion of collateral circulation play a central role in the pathogenesis of PHT. PHT is also characterized by changes in vascular structure, termed vascular remodeling, which is an adaptive response of the vessel wall that occurs in response to chronic changes in the environment such as shear stress. Angiogenesis, the formation of new blood vessels, also occurs with PHT related in particular to the expansion of portosystemic collateral circulation. The complementary processes of vasoreactivity, vascular remodeling, and angiogenesis represent important targets for the treatment of portal hypertension. Systemic and splanchnic vasodilatation can induce hyperdynamic circulation which is related with multi-organ failure such as hepatorenal syndrome and cirrhotic cadiomyopathy.     

大鼠肝前性门静脉高压症形成中一氧化氮和内皮素-1水平的动态变化

目的:观察大鼠肝前性门静脉高压症形成中一氧化氮(NO)和内皮素-1(ET-1)水平的动态变化,探讨其在门静脉高压症高动力循环中的作用。方法:以部分门静脉结扎(PVL)法复制肝前性门静脉高压症大鼠模型,分别用硝酸还原酶和放免法检测正常组、假手术(SO)组及PVL组术后不同时点的门静脉血浆NO^-₂/NO^-₃、ET-1水平,并同步监测血流动力学指标的动态变化。结果:PVL术后各时点NO^-₂/NO^-₃水平显著高于而ET-1水平显著低于正常组,同时伴有血流动力学的明显变化。结论:门静脉高压症大鼠存在高动力循环状态(HCS)。NO和ET-1参与HCS的形成和维持。     

Modifications produced by indomethacin and L-NAME in the effect of ultralow-dose aspirin on platelet activity in portal hypertension

In our previous study, we demonstrated the effect of ultralow-dose aspirin (ULDA) on platelet activity and bleeding in rats with portal hypertension (PHT) produced by portal vein ligation (PVL). This paper reports modifications in this effect caused by blocking NO production by nitro arginine methyl ester (NAME) and cyclooxygenase (COX) activity with indomethacin. PVL rats and sham-operated controls were treated with placebo, indomethacin or NAME and 30 min thereafter with placebo or ULDA treatment. Platelet activity was studied by a model of in vivo laser-induced thrombus production in the mesenteric circulation, induced hemorrhage time (IHT) and platelet aggregation ex vivo induced by adenosine diphosphate in an aggregometer. The PVL group receiving placebo showed a decreased platelet activity with prolonged IHT, an effect that was reversed by ULDA. Indomethacin induced a decreased platelet activity in the control rats and a prolonged IHT. In PHT with ULDA, in vivo platelet activity was enhanced but the normalization of IHT observed in rats without indomethacin was blunted. The addition of NAME normalized the diminished in vivo platelet aggregation and increased the IHT observed in PVL animals. These changes decreased the effect of ULDA in both sham-operated and PVL animals. The effect of indomethacin was more clearly modified by ULDA than the effect of NAME, thus suggesting that modifications in the COX pathway might alter the effect of ULDA. The simultaneous administration of indomethacin and ULDA could inhibit its beneficial effect on bleeding in rats with PHT.     

Hemodynamic Changes in Splanchnic Blood Vessels in Portal Hypertension

Portal hypertension (PHT) is associated with a hyperdynamic state characterized by a high cardiac output, increased total blood volume, and a decreased splanchnic vascular resistance. This splanchnic vasodilation is a result of an important increase in local and systemic vasodilators (nitric oxide, carbon monoxide, prostacyclin, endocannabinoids, and so on), the presence of a splanchnic vascular hyporesponsiveness toward vasoconstrictors, and the development of mesenteric angiogenesis. All these mechanisms will be discussed in this review. To decompress the portal circulation in PHT, portosystemic collaterals will develop. The presence of these portosystemic shunts are responsible for major complications of PHT, namely bleeding from gastrointestinal varices, encephalopathy, and sepsis. Until recently, it was accepted that the formation of collaterals was due to opening of preexisting vascular channels, however, recent data suggest also the role of vascular remodeling and angiogenesis. These points are also discussed in detail. Anat Rec, 291:699–713, 2008. © 2008 Wiley‐Liss, Inc.     

Hepatotrophic factors in liver growth and atrophy.

The hepatotrophic effect of portal blood was studied in rats with portal branch ligation (PBL) simultaneously subjected to mesocaval shunt or total hepatic arterialization. A direct supply of the nonligated liver lobes with pancreatic effluent did not improve DNA synthesis or mitotic response compared to arterialized rats, in which the portal-borne hepatotrophic factors reached the same lobes after systemic recirculation. A comparison with shunted but Sham-PBL rats showed that restorative capacity of the liver was not seriously impaired. In rats with PBL alone the lobes with portal occlusion showed extensive necrosis. In contrast, in PBL + shunt rats necrosis was significantly inhibited, indicating a hepatoprotective role of portal blood during hepatic arterial recirculation. Thus, the study suggests that quality of hepatic blood supply is of vital importance in maintenance of hepatocellular integrity. Hemodynamical factors seem to be of importance, but more in a sense of increased or decreased accessibility of humoral hepatotrophic factors in the blood.     

Hepatotrophic factors in liver growth and atrophy

The hepatotrophic effect of portal blood was studied in rats with portal branch ligation (PBL) simultaneously subjected to mesocaval shunt or total hepatic arterialization. A direct supply of the nonligated liver lobes with pancreatic effluent did not improve DNA synthesis or mitotic response compared to arterialized rats, in which the portal-borne hepatotrophic factors reached the same lobes after systemic recirculation. A comparison with shunted but Sham-PBL rats showed that restorative capacity of the liver was not seriously impaired. In rats with PBL alone the lobes with portal occlusion showed extensive necrosis. In contrast, in PBL + shunt rats necrosis was significantly inhibited, indicating a hepatoprotective role of portal blood during hepatic arterial recirculation. Thus, the study suggests that quality of hepatic blood supply is of vital importance in maintenance of hepatocellular integrity. Hemodynamical factors seem to be of importance, but more in a sense of increased or decreased accessibility of humoral hepatotrophic factors in the blood.     

On the Mechanisms Responsible for Selection of Hepatic Veins as Target for Thrombosis Following Injection of Endotoxin in Hyperlipemic Rats

The feeding of a butter-rich diet, to sensitize rats for studying the phenomenon of hepatic vein thrombosis, is shown to produce severe liver steatosis leading to a sinusoidal barrage and portal hypertension. The portal pressure in these animals was 210 ± 4 mm of saline, as compared to 113 ± 3 mm in the normal rat. Blood circulation studies using carbon suspensions revealed production of a vascular stasis in the hepatic veins after 60 to 90 minutes, when endotoxin (Salmonella typhosa, 0.3 mg/kg) is introduced into the blood circulation to initiate hepatic vein thrombosis. Similar results were observed after 15 minutes with ellagic acid (1 mg/kg/min). The stasis was found in connection with an additional intrahepatic resistance to blood flow as evidenced by a rise in portal pressure and by a reduction in liver perfusion in relation with development of systemic hypotension. In contrast with this, endotoxin initiated only slight and transient changes in the normal rat. Thrombosis immediately followed production of stasis in the hepatic vein, whether the phenomenon was initiated by endotoxin or ellagic acid. Furthermore, inhibition of the vascular stasis of α-adrenergic blockade (phenoxybenzamine, 3 mg/kg) was accompanied by prevention of hepatic vein thrombosis. It is concluded that stasis in the hepatic veins resulting from a mechanical obstruction of the circulation by steatosis and by an additional reduction in blood flow initiated by endotoxin, is responsible for selection of hepatic veins as targets for thrombosis following injection of endotoxin in hyperlipemic rats.     

The mast cell integrates the splanchnic and systemic inflammatory response in portal hypertension

Portal hypertension is a clinical syndrome that is difficult to study in an isolated manner since it is always associated with a greater or lesser degree of liver functional impairment. The aim of this review is to integrate the complications related to chronic liver disease by using both, the array of mast cell functions and mediators, since they possibly are involved in the pathophysiological mechanisms of these complications. The portal vein ligated rat is the experimental model most widely used to study this syndrome and it has been considered that a systemic inflammatory response is produced. This response is mediated among other inflammatory cells by mast cells and it evolves in three linked pathological functional systems. The nervous functional system presents ischemia-reperfusion and edema (oxidative stress) and would be responsible for hyperdynamic circulation; the immune functional system causes tissue infiltration by inflammatory cells, particularly mast cells and bacteria (enzymatic stress) and the endocrine functional system presents endothelial proliferation (antioxidative and antienzymatic stress) and angiogenesis. Mast cells could develop a key role in the expression of these three phenotypes because their mediators have the ability to produce all the aforementioned alterations, both at the splanchnic level (portal hypertensive enteropathy, mesenteric adenitis, liver steatosis) and the systemic level (portal hypertensive encephalopathy).     

The responses of the hepatic and splanchnic vascular beds to vasopressin in rats

Vasopressin, a vasoactive peptide, causes vasoconstriction via V1a vasopressin receptors. Unlike other vasoconstrictor agents, vasopressin also has vasodilatory properties. The purpose of this study was to determine the effect of vasopressin on hepatic and splanchnic circulation in Sprague- Dawley rats. The experiments were conducted in not only isolated blood- and constant flowperfused livers but also anesthetized spontaneously breathing rats. In anesthetized rats, portal venous pressure (Ppv), systemic arterial pressure (Psa), central venous pressure, and hepatic blood flow (HBF) of combined portal venous and hepatic arterial blood flow were continuously measured, and splanchnic vascular bed resistance (Rspl) defined by (Psa - Ppv) / HBF was determined. In perfused livers, vasopressin at 0.1-1,000 nM caused weak venoconstriction as evidenced by small increase in Ppv. In anesthetized rats, when vasopressin was injected into the portal vein as a bolus consecutively at 0.01-100 nmol/kg, Psa increased dose-dependently with the peak increment of 60 ± 18 mmHg at 100 nmol/kg. Ppv and HBF decreased, with resultant increase in Rspl, indicating splanchnic vasoconstriction. In conclusion, hepatic venoconstrictor action of vasopressin was weak in rats. Vasopressin causes splanchnic vasoconstriction, resulting in a decrease in HBF and Ppv in anesthetized rats.     

血吸虫病性门静脉高压症兔肝脏微血管构筑变化的研究

目的:探讨血吸虫病门静脉高压症时肝脏微血管构筑的变化及其可能在全身高动力循环状态中的作用。方法:采用腹部敷贴法感染血吸虫尾蚴建立血吸虫病性肝纤维化模型,经插管检测心输出量(CO)、平均动脉压(MAP)、心率(HR)和肝静脉嵌塞压(WHVP),按公式计算心脏指数(CI)、外周血管阻力(SVR);通过血管铸型方法观察肝脏微血管构筑。结果:与正常兔比较,血吸虫病兔CO、CI明显增高,MAP和SVR显著降低,WHVP升高,两组间HR差异无统计学意义。肝脏微血管铸型观察,血吸虫病时肝内微血管形态和比例严重失常,肝窦显著膨大,门静脉主干增粗,肝内形成广泛的小吻合支,其间以门静脉终末支与肝静脉终末支、门静脉小分支直接引流入肝静脉多见。结论:血吸虫病性门静脉高压症兔存在肝内门-体分流病理改变,可能是形成全身高动力循环状态并维持门静脉高压的一个重要环节。