乙肝特异性人胎盘转移因子转移皮肤迟发型超敏反应的实验研究

研究证明,注入乙型肝炎病毒特异性人胎盘转移因子的实验小鼠(A组)对乙型肝炎病毒疫苗可产生明显的皮肤迟发型超敏反应,与注入生理盐水的对照组(C组)比较,有非常显著性差异(P<0.001),而注入正常人胎盘转移因子的小鼠(B组),对乙型肝炎病毒疫苗也可产生皮肤退发型超敏反应,但不及A组明显。实验表明从乙型肝炎病毒表面抗体阳性人胎盘中提取的乙型肝炎病毒特异性人胎盘转移因子可转移抗原特异性皮肤迟发型超敏反应。     

高免卵黄透析液转移迟发型超敏反应的实验研究

建立一种新的制备特异性转移因子的方法。方法：用二硝基氯苯（ＤＮＣＢ）免疫鸡,当观察到迟发型超敏反应发生后,收集鸡蛋制备ＤＮＣＢ高免卵黄透析液,并将其注入小鼠腹腔内,６ ｄ后用 ＤＮＣＢ攻击小鼠并观察其迟发型超敏反应发生的情况。结果：ＤＮＣＢ高免卵黄透析液组、对照卵黄透析液组和生理盐水组的皮肤反应（Ａ值）分别为０．０５２±０．０２２、０．０２８±０．０１４和０．０３２±０．０１２（Ｐ＜０．０５）。结论：高兔卵黄透析液含有某些介导细胞免疫的小分子物质。     

天螺霜对免疫功能低下小鼠模型特异性免疫功能的影响

目的 研究天螺霜作为免疫增强剂对免疫功能低下模型小鼠特异性免疫功能的影响及其剂量效果关系。方法 采用皮下注射环磷酰胺制造小鼠免疫功能低下模型，观察并比较不同剂量天螺霜（400mg／kg、200mg／kg、50mg／kg）与环磷酰胺配合使用对以鸡红细胞为抗原的溶血素生成和二硝基氯苯所致小鼠迟发型超敏反应的影响。结果 模型组小鼠溶血素、溶血空斑形成和迟发型超敏反应均明显下降．而阳性药物对照组和高、中剂量天螺霜组该免疫功能均明显增强（P〈0．05或P〈0．01），且其作用强度呈现剂量依赖性，高剂量组作用略强于阳性药。结论 天螺霜可明显增强免疫功能低下模型小鼠特异性免疫功能．作用呈剂量依赖性。最高剂量推荐200～400mg／kg。     

肺癌特异性转移因子口服液的研制

目的制备羊脾特异性转移因子口服液,并检测其免疫活性。方法采用肺癌患者术后肿瘤组织加佐剂免疫羊,取其脾用透析法制备肺肿瘤特异性转移因子。检测其理化性质,并通过E玫瑰花环试验、白细胞粘附抑制法(LAI)和迟发型皮肤超敏反应实验(DTH)检测特异免疫活性。结果制品的各项理化指标均达到了国家药品标准(WS1-XG-037-2000),E-玫瑰花法结花率为20.5%,多肽含量为1.38 mg/mL,核糖含量为83.5μg/mL。该制品能明显抑制小白鼠白细胞黏附,并能诱导小鼠皮肤迟发型超敏反应,表明该口服液对T淋巴细胞的增殖和玫瑰花结的形成均有促进作用。结论本法制备的羊脾特异性转移因子口服液,为抗肿瘤制剂的研究奠定理论基础。     

新型乙型肝炎病毒表面抗原中蛋白核酸疫苗的免疫原性研究

目的 观察新型乙型肝炎病毒（HBV）表面抗原中蛋白（MHBs)核酸疫苗的免疫原性。方法 以新型人体应用载体质粒pSW3891构建MHBs核酸疫苗（pSW3891/MHBs/adr)，对照组和实验组Balb/c小鼠分别基因枪法免疫对照载体质粒（pSW3891)和MHBs核酸疫苗，采用酶联免疫吸附试验检测抗HBs,LDH释放测定法检测小鼠特异性细胞毒T淋巴细胞（CTL）杀伤活性。结果 MHBs核酸疫苗可在体外高效表达，免疫小鼠后可产生高滴度抗HBs，血清阳性终点滴度达1：97200,小鼠脾细胞HBs特异性细胞毒性T淋巴细胞（CTL）杀伤活性达78.81%。结论 新型MHBs核酸疫苗在Balb/c小鼠实验中表现出良好的体液和细胞免疫原性。     

对HIV-1 env DNA疫苗免疫应答的遗传学控制

含有编码1型人类免疫缺陷症病毒(HIV-1)env蛋白的HIV-1特异性DNA疫苗可诱导强的体液和细胞介导免疫应答。作者用不同品系的小鼠研究了HIV-1特异性DNA疫苗诱导抗体产生及迟发型超敏反应(DTH)的遗传学控制。     

小■胺对小鼠迟发型超敏反应的影响(英文)

本文研究了小檗胺对小鼠由绵羊红细胞，二硝基氯苯及同种异型脾细胞所诱导的迟发型超敏反应的影响。实验结果显示小檗胺能明显抑制小鼠的迟发型超敏反应，表明小檗胺可能对小鼠的细胞免疫功能具有抑制作用     

乙型肝炎病毒特异性转移因子的研究

制备一种乙型肝炎病毒特异性转移因子制剂(HBV-STF),为临床应用提供有价值的实验依据.从人HBsAg阳性胎盘中制备了HBV-STF,并对其理化性质、免疫学活性进行了检测和初步的临床试用.每批样品经无菌试验、热原质检查、动物安全性试验等均符合药典要求.本品最大紫外吸收光谱在256±2 nm处,E260nm/E280nm比值大于2.7.其水解氨基酸含17种.对人T淋巴细胞E受体的激活试验结果显示,HBV-STF的Ea-RFc平均增高率在83.47%～103.48%之间,抗原特异性皮肤试验表明HBV-STF能刺激小鼠体内T淋巴细胞增殖,诱导小鼠跖趾部皮肤的迟发性变态反应.对HBV-STF的初步临床试用也取得明显效果,显示HBV-STF是一种可用于治疗乙肝的安全、有效的免疫调节剂.     

转移因子在移植免疫中的作用

转移因子参与移植免疫的迟发型超敏反应。业已证明,移植皮肤4～6天后就有转移因子产生,并保持其活性达40天。以不同方法,包括皮内法给动物注射转移因子能改变移植物排斥速率。注射纯转移因子能加     

10－羟基－2－癸烯酸对小鼠T淋巴细胞及其亚型和白介素2产生的影响

观察了１０－羟基－２－癸烯酸（１０－ＨＤＡ）对小鼠Ｔ淋巴细胞及其亚型和白介素２产生的影响．结果表明，１０－ＨＤＡ１，５，２５ｍｇ·ｋｇ－１·ｄ－１ｉｐ，５ｄ可拮抗环磷酰胺１００ｍｇ·ｋｇ－１对小鼠迟发型超敏反应（ＤＴＨ）的抑制作用．体外给药，１０－ＨＤＡ可促进刀豆球蛋白Ａ诱导的Ｔ淋巴细胞增殖反应；促进小鼠脾细胞产生白介素２．采用单克隆抗体间接免疫荧光法证明１０－ＨＤＡ可增加小鼠胸腺Ｌ３Ｔ＋４细胞数，而对Ｌｙｔ－２＋细胞无明显影响．结果提示１０－ＨＤＡ可调节Ｔ淋巴细胞参与的免疫反应．     

孕期注射乙肝免疫球蛋白阻断宫内与胎盘感染的相关性研究

目的探讨HBsAg阳性孕妇孕期注射乙肝免疫球蛋白（HBIG）阻断母婴传播的疗效与胎盘HBV感染之间的相关性。方法92例HBsAg阳性孕妇分成两组,研究组44例,于妊娠28、32及36周肌注HBIG 200IU;对照组48例,未用药。采用荧光定量PCR及ELISA法．分别检测孕妇及其新生儿静脉血HBVDNA水平及乙肝五项。采用免疫组化S—P法,检测胎盘各层细胞HBsAg及HBcAg的表达。结果（1）92例孕妇胎盘HBV感染43例,各层细胞感染后导致官内传播的相对危险度（OR值）由母面至胎儿面呈逐渐上升趋势。（2）研究组胎盘HBV总的感染率为34．09％（15/44）．而对照组为58-33％,两组比较,差异有显著性（P〈0．05）。（3）研究组胎盘VCEC感染：率为6．82％（3／44）,而对照组为27．08％（13／48）。两组比较,差异有显著性（P〈0．05）。结论（1）胎盘VCECHBV感染是官内感染的高危因素之一。（2）注射HBIG可降低胎盘HBV感染率。     

体外免疫法制备猪脾乙肝特异性转移因子

以乙肝疫苗为抗原,猪脾细胞为效应细胞,进行体外免疫反应,成功地制备出猪脾乙肝特异性转移因子PS_（HBV）-TF.抗原特异性皮肤试验证实,该PS_（HBV）-TF具有转移抗乙肝病毒特异性细胞免疫功能.     

慢性乙型肝炎产妇胎盘组织丝裂原激活蛋白激酶表达与乙型肝炎病毒宫内传播的研究

目的通过观察孕妇胎盘组织的丝裂原激活蛋白激酶（MAPK）表达与乙型肝炎病毒（HBV）持续感染的关系,探讨细胞内信号变化与HBV宫内传播分子机制。方法研究对象为2010年1月至2011年11月在我院自然分娩或剖宫产的慢性乙型肝炎产妇52例,分为HBeAg阳性组（22例）和HBeAg阴性组（25例）,5例肝功能正常、HBV和丙型肝炎病毒（HCV）血清标志物均为阴性的产妇为正常对照组。均于分娩后迅速无菌收集胎盘组织,应用定量逆转录聚合酶链反应（RT—PCR）方法检测胎盘组织中MAPKmRNA水平,并分别定量检测其血清HBV—DNA和HBeAg水平。结果HBeAg阳性组HBV—DNA水平高于HBeAg阴性组,组间差异有统计学意义[（0．6±0．5）×10^6拷贝／L比（4．6±2．1）×10^6拷贝／L,P〈0．01]。正常对照组、HBeAg阴性组和阳性组的MAPK基因表达分别为0．98、0．70、0．56,3组之间分别两两比较,差异均有统计学意义（均P〈0．05）。结论胎盘组织中MAPK的表达与其持续感染HBV有一定关系。     

HLA-DRB1基因多态性与HBV宫内感染的相关性

目的 探讨HLA-DRBl等位基因多态性与HBV宫内感染的关系.方法 选择HBsAg阳性孕妇分娩的新生儿中发生HBV宫内感染的母亲21例(A组),而未发生宫内感染的母亲46例(B组)以及正常母亲42例(C组),采用聚合酶链反应-序列特异性引物(PCR-SSP)技术扩增HLA-DRB1四对等位基因,计算各刑别出现频率.结果 (1)HLA-DRB1*11在A组的等位基因频率明显高于B组(38.1% vs 13.0%)(P<0.05);(2)HLA-DRB1*13、HLA-DRB1*03在A、B两组的等位基因频率明显低于C组(4.5% vs 33.3%,9.0%vs 28.6 0A)(P<0.01):(3)HLA-DRB1*07等位基因频率在A、B、C三组间差异无显著性.结论 HLA-DRB1*11与HBV宫内感染有关;HLA-DRB1*03、13可能与HBV清除有关.     

乙型肝炎病毒基因型分布及临床意义

目的 了解珠海地区乙型肝炎病毒(HBV)基因型流行病学分布特征,并分析不同基因型与HBV感染谱的相关性.方法 采用聚合酶链反应-限制片段长度多态性(PCR-RFLP)的基因型分型方法对62例HBV携带者及63例慢性乙型肝炎、7例慢性重症乙型肝炎、5例肝硬化患者感染的HBV进行分型,并分析不间感染谱中HBV基因型的分布.结果 137例感染者HBV基因型B型72例(52.6%),C型65例(47.4%),慢性肝炎患者、重症肝炎、肝硬化患者的C基因型比例[45例(69.2%)、5例(7.7%)、4例(6.2%)]明显高于B基因型[18例(25.0%)、2例(2.8%)、1例(1.4%)],差异均有统计学意义(均P＜0.05);HBV携带者B基因型比例[51例(70.8%)]明显高于C基因型[11例(16.9%)],差异有统计学意义(P＜0.05).C基因型慢性肝炎患者的HBV-DNA、平均ALT水平高于B基因型慢性肝炎患者,但差异均无统计学意义(P＞0.05).结论 珠海地区HBV感染者以B型和C型为优势基因型,C基因型可能是乙型肝炎病情进展的相关因素.

Abstract：

Objective To understand the distribution of hepatitis B virus (HBV) genotypes in Zhuhai area and to analyze the relationship between HBV genotypes and patients with hepatitis B. Methods A polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis assay amplifying S-segment sequences of HBV-DNA were used for analyzing the distribution of HBV genotypes among 62 HBV carriers (ASC group),63 patients with chronic hepatitis B (CHB group),7 patients with severe chronic hepatitis B (CSH group)and 5 cases liver cirrhosis (LC group). Results The proportion of HBV genotyjpes B and C was 52.6% and 47.4%. There were no significant differences of indexes related to liver function damage (ALT) or serum HBV DNA levels between subtype B and subtype C infected patients. Conclusion The genotypes B and C are two predominant genotypes in Zhuhai area;genotype C may be associated with the development of hepatitis.     

不同来源的抗乙肝免疫核糖核酸对慢性乙型肝炎患者的免疫调节作用

目的 探讨HBV感染后乙肝保护抗体抗－HBs产生机体外周淋巴细胞iRNA(h-iRNA)对慢性乙型肝炎的免疫调节作用,并与目前单用HBsAg免疫动物来源的抗乙肝iRNA(a-iRNA)相比较。方法 利用四甲基偶氮唑盐（MTT）法观察h-iRNA对慢性乙肝患外周淋巴细胞HBV抗原特增殖反应的影响。结果 在HBsAg刺激组中,h-iRNA,a-iRNA对慢性乙肝患外周HB－sAg特异淋巴细胞增殖反应皆有一定程度的增强作用;但HBcAg刺激组中,h-iRNA对慢性乙肝患HBc-Ag特异淋巴细胞增殖反应具有增强作用,与a-iRNA相比,差异有显性意义。结论 HBV抗原的特异增殖反应,对主要靶抗原HBcAg特异淋巴细胞增殖反应的增强作用,有利于清除肝内HBV。目前单用HBsAg免疫动物的抗乙肝iRNA,不具有对HBV其它抗原的免疫信息,尤其是不能对免疫细胞识别肝细胞内HBV的主要靶抗原HBcAg产生有效免疫作用,可能是影响其疗效的重要原因。     

Placental transfer and DNA binding of benzo(a)pyrene in human placental perfusion

Benzo(a)pyrene (BP) is the best studied polycyclic aromatic hydrocarbon, classified as carcinogenic to humans. The carcinogenic metabolite, benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE), binds covalently to DNA. The key enzyme in this metabolic reaction is CYP1A1, which has also been found in placenta and human trophoblastic cells. By using human placental perfusion we confirmed that BP added to the maternal circulation in concentrations of 0.1 and 1 μM reaches fetal compartment but somewhat slower than the freely diffusible reference substance antipyrine. A well-known P-glycoprotein (ABCB1/P-gp) antagonist verapamil did not affect the transfer more than it did in the case of antipyrine, indicating that ABCB1/P-gp does not have a role in BP transfer. In one of the two placentas perfused for 6 h with the higher concentration of BP (1 μM) BPDE specific DNA adducts were found in placental tissue after the perfusion, but not before. The ability of human trophoblastic cells to activate BP to BPDE–DNA adducts was confirmed in human trophoblastic BeWo cells. This study shows that maternal exposure to BP leads to the exposure of the fetus to BP and/or its metabolites and that placenta itself can activate BP to DNA adducts.     

Organic anion transporting polypeptide 2B1 and breast cancer resistance protein interact in the transepithelial transport of steroid sulfates in human placenta.

The human placenta has both protective and nurturing functions for the fetal organism. Uptake and elimination of xenobiotics and endogenous substances are facilitated by various transport proteins from the solute carrier (SLC) and ABC families, respectively. A functional interaction of uptake and elimination, which is a prerequisite for vectorial transport across cellular barriers, has not been described for placenta. In this study, we examined expression of organic anion transporter (OAT) 4 (SLC22A11), organic anion transporting polypeptide (OATP) 2B1 (SLCO2B1, OATP-B), and breast cancer resistance protein (BCRP) (ABCG2) in human placenta (n = 71) because all three proteins are involved in transmembranal transfer of estrone 3 sulfate (E3S; metabolic product) and dehydroepiandrosterone sulfate (DHEAS; precursor molecule). On the mRNA level, we found a significant correlation of OATP2B1 and BCRP (R(2) = 0.534; p < 0.01) but not between OAT4 and BCRP (R(2) = -0.104; p > 0.05). Localization studies confirmed basal expression of OATP2B1 and apical expression of BCRP. To study functional interactions between OATP2B1 and BCRP, we developed a Madin-Darby canine kidney cell model expressing both transport proteins simultaneously (OATP2B1 and BCRP in the basal and apical membrane, respectively). Using this cell model in a transwell system resulted in a significantly increased basal to apical transport of both E3S and DHEAS, when both transporters were expressed with no change of transfer in the apical to basal direction. Taken together, these data show the potential for a functional interaction of OATP2B1 and BCRP in transepithelial transport of steroid sulfates in human placenta.     

乙型肝炎患者血清HBV前C区A83变异株的检测

目的:探讨乙型肝炎病毒(HBV)前C区A83点突变与临床关系。方法:采用聚合酶链反应(PCR)对64例乙型肝炎(乙肝)患者HBV前C区基因进行扩增,对PCR产物进行限制性片段长度多态性分析(RFLP),鉴定前C区第83位核苷酸点突变。结果:34例(53.13%)有A83点突变,其中12例(18.75%)为单独变异株感染,22例(34.38%)为变异株与野毒株混合感染。慢性乙型肝炎(CHB)和肝硬化(LC)组高于无症状HBV携带者(ASC)(P<0.05)。抗-HBe( )组的检出率为70.0%,高于HBeAg( )组的25.0%(P<0.01),并且随着HBVDNA含量的增高,其发生变异的可能性越大(P<0.05)。结论:前C区A83点突变的发生可能与机体长期的炎症活动、免疫压力的选择有关。     

The effect of probenecid and MK-571 on the feto-maternal transfer of saquinavir in dually perfused human term placenta

Human placenta, particularly the blood–placenta barrier, with various transporters has crucial role to protect the fetus and, on the other hand, to facilitate movement of compounds towards the fetal circulation. This study aimed to characterize the role of basal transporters of the syncytiotrophoblast, which appear to be yet less studied, in the fetal-to-maternal transfer of saquinavir by use of dually perfused human placentas.A dual perfusion of human placenta was performed to study effect of MK-571 and probenecid, inhibitors of the MRP1 and OATP transporters, expressed in the basal trophoblast membrane, on the transfer of saquinavir. The fetal-to-maternal placental transfer of saquinavir in the control group as measured by TPT_AUC% (absolute fraction of the dose crossing placenta) was 14.0%, which is 73% less than the transfer of the freely diffusible antipyrine. The two inhibitors, MK-571 and probenecid caused a non-significant (P = 0.34 for ANOVA) reduction of 43% and 24%, respectively, in the mean amount of saquinavir transferred from the fetal to the maternal side. MK-571 also somewhat (by 31%) reduced the TPT_AUC% of antipyrine, but this finding did not reach statistical significance (P = 0.25). Neither of the employed inhibitors had an effect on the placental transfer index of saquinavir transfer (P = 0.77). The present results indicated lack of significant effect by MK-571 and probenecid on the fetal-to-maternal transfer of saquinavir and suggest that MRP1 and, possibly, OATP2B1 do not play a significant role in the fetal-to-maternal transfer of saquinavir.