氟化钠致大鼠肾脏脂质过氧化损伤及亚硒酸钠的保护作用

目的研究氟化钠（ＮａＦ）对大鼠肾脏脂质过氧化作用的影响及亚硒酸钠（Ｎａ２ＳｅＯ３）对氟肾脏毒性的保护作用。方法大鼠经饮水加入ＮａＦ（１５ｍｍｏｌ／Ｌ）或／和Ｎａ２ＳｅＯ３（１５μｍｏｌ／Ｌ）染毒１２０天,观察氟在机体的蓄积、排泄,尿酶活性及肾脏脂质过氧化水平的变化。结果氟染毒后,大鼠血氟水平、尿氟水平、骨氟含量,尿谷氨酰转肽酶（γＧＴ）、Ｎ乙酰βＤ氨基葡萄糖苷酶（ＮＡＧ）、琥珀酸脱氢酶（ＳＤＨ）活性及肾脏活性氧自由基（ＯＦＲＳ）相对浓度、脂质过氧化降解产物丙二醛（ＭＤＡ）含量均显著升高,谷胱甘肽过氧化物酶（ＧＳＨＰｘ）活性及ＧＳＨ含量显著下降。同时补加亚硒酸钠,可促进尿氟排泄,降低血氟水平及骨氟含量,降低尿酶γＧＴ、ＮＡＧ、ＳＤＨ活性和肾脏ＯＦＲＳ、ＭＤＡ含量,同时提高肾脏ＧＳＨＰｘ活性,但对ＧＳＨ含量影响不显著。结论氟化钠引起肾脏损伤与其诱导脂质过氧化作用增强有关,亚硒酸钠对氟化钠致肾脏毒性具有拮抗作用。     

延胡索酸二甲酯对大鼠醌还原酶和谷胱甘肽—S—转移酶的诱导作用

目的　探讨延胡索酸二甲酯（ＤＭＦ） 对大鼠主要脏器醌还原酶和谷胱甘肽Ｓ转移酶的诱导作用和意义。方法　雄性Ｗｉｓｔａｒ 大鼠饮食中补充０ ．２ ％ ＤＭＦ,于６ 、２４ 、７２ 小时、１ 和２ 周后取血并处死动物,用酶动力学法测定大鼠腺胃、肝、肺、肾和心脏中的醌还原酶（ＱＲ） 和谷胱甘肽Ｓ转移酶（ＧＳＴｓ） 活性,并按此法测定血清ＱＲ、ＧＰＴ和乳酸脱氢酶含量。结果　服用ＤＭＦ２４ 、７２ 小时、１ 和２ 周后可观察到腺胃、肝、肺、肾脏中ＱＲ 和ＧＳＴｓ 活性显著增高（ Ｐ＜ ０ ．０００ １ 或Ｐ＜ ０ ．０１） ,同时血清ＱＲ 活性也显著增高（ Ｐ＜ ０ ．０１ ,或Ｐ＜ ０ ．０００ １） 。ＤＭＦ对血清ＧＰＴ 和乳酸脱氢酶的活性则无影响。结论　ＤＭＦ对大鼠腺胃、肝、肺、肾和血清中的ＱＲ 和ＧＳＴｓ 活性具有诱导作用,ＤＭＦ有望成为人类抗氧化损伤的保护剂和解毒剂。     

拟除虫菊酯对大鼠脑组织及肝脏生物转化酶的影响

目的 实验研究溴氰菊酯（ＤＭ）和氯菊酯（ＰＭ）对大鼠脑组织及肝脏某些生物转化酶的影响。方法 应用荧光分光光度法、二硝基苯肼比色法和Ｈａｂｉｇ法检测７－乙氧基试卤灵－Ｏ－脱乙基酶９ＥＲＯＤ０、Ｂ型单胺氧化酶９ＭＡＯＢ）和谷胱甘肽Ｓ－转移酶（ＧＳＴ）活力。结果 整体实验中，ＤＭ染毒动物肝脏ＥＲＯＤ、ＭＡＯＢ活力分别下降了４７．９４％和２２．７６％；脑组织和肝脏ＧＳＴ活力分别增加了２２．２０％和３９．５     

甲基叔丁基醚对大鼠肝组织基因表达的影响

目的甲基叔丁基醚（ＭＴＢＥ）是一种新型的汽油添加剂,动物试验显示其具有一定的致癌性。为了解其动物致癌性的可能机制,我们检测了经ＭＴＢＥ亚慢性染毒的大鼠肝组织中原癌基因ｃｍｙｃ基因和功能基因谷胱甘肽Ｓ转移酶Ｐ（ＧＳＴＰ）基因的表达情况。方法４０只雄性ＳＤ大鼠,体重１８０～２００ｇ,随机分为４组。将ＭＴＢＥ溶于适量豆油中,灌胃染毒,染毒剂量分别为２００ｍｇ／ｋｇ,６００ｍｇ／ｋｇ,１０００ｍｇ／ｋｇ和对照组。每天１次,每周５天,共１３周。动物肝组织于液氮速冻后,一步法提取总ＲＮＡ。随机引物法地高辛标记ｃｍｙｃ和ＧＳＴＰ探针,与ＲＮＡ进行点杂交,图像分析仪分析结果。结果大鼠肝组织中ｃｍｙｃ基因表达水平明显增高,ＧＳＴＰ基因表达未见增强。结论ＭＴＢＥ可明显诱导ｃｍｙｃ基因的高表达,提示其具有促进细胞增殖的作用,这是其动物致癌性的可能机制之一。     

还原型谷胱甘肽对急性百草枯中毒治疗作用的实验研究

探讨还原型谷胱甘肽对急性百草枯中毒的治疗作用。方法在大鼠用百草枯（Ｐａｒａｑｕａｔ,ＰＱ）灌胃（２５０ｍｇ／ｋｇ）染毒后不同时间腹腔注射还原型谷胱甘肽（ＧＳＨ）,分别测定染毒后８ｈ、２４ｈ、４８ｈ,７２ｈ大鼠血浆和支气管－肺泡灌洗液（ＢＡＬＦ）中丙二醛（ＭＤＡ）含量及超氧化物歧化酶（ＳＯＤ）、谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）的活力,并观察肺组织结构改变。结果ＰＱ染毒后血浆及ＢＡＬＦ中ＭＤＡ显著高     

实验性煤矽肺大鼠氧化和抗氧化损伤的动态观察

大鼠经气管注入煤石英尘，观察肺泡巨噬细胞（ＡＭ）和支气管肺泡冲洗液（ＢＡＬＦ）中脂质过氧化产物和抗氧化物质的动态变化。结果表明，染尘后７天ＡＭ中ＭＤＡ含量增加，ＳＯＤ活性下降，以后恢复至正常，ＧＳＨＰｘ活性和ＧＳＨ含量染尘后７～３０天升高。ＢＡＬＦ中ＭＤＡ含量、ＳＯＤ活性无变化，ＧＳＨＰｘ活性、ＧＳＨ含量均低于石英组。说明煤石英尘诱发ＡＭ脂质过氧化反应发生于染尘初期，且明显弱于石英尘。说明脂质过氧化反应在煤矽肺病变形成中不起主要作用     

急性染镉对小鼠肝肝肾组织谷胱甘肽的影响

目的 探讨氯化镉急性染毒对小鼠肝脏、肾脏胱甘肽（ＧＳＨ）代谢的影响。方法 给小鼠一次性腹腔注射不同剂量氯化镉,９小时后测定肝,肾组织镉、丙二醛（ＭＤＡ）、ＧＳＨ含量。结果 各剂量组肝、肾镉含量升高,高剂量组肝、肾ＭＤＡ含量升高,肾ＧＳＨ含量下降,肝脏ＧＳＨ呈现先升高,后下降的剂量反应关系,。结论氯化镉染毒９小时后可同时在小鼠肝、肾脏蓄积、并引起肝肾脂质过氧化反应,小鼠肝、肾脏对急性镉染毒诱导的ＧＳ     

绿茶及混合茶抑制二乙基亚硝胺诱发大鼠肝癌前期病变的抗氧化机理研究

用二乙基亚硝胺（ＤＥＮ）诱发Ｗｉｓｔａｒ大鼠肝癌前期病变（Ｓｏｌｔ－Ｆａｒｂｅｒ模型）。实验前６周给大鼠饮用２％绿茶水及０．５％混合茶水，于实验第８周末宰杀动物。结果发现，绿茶组及混合茶组肝ｒ－ＧＴ异常病灶数均明显低于阳性对照组（Ｐ＜０．０１）。经ＨＰＬＣ法分离肝微粒体脂质过氧化产物丙二醛（ＭＤＡ）、乙醛（ＡＣＴ）、戊醛（ＰＰ）、丙酮（ＡＣＯＮ），与阳性对照组相比，绿茶组和混合茶组的各种脂质过氧化产物均有不同程度的降低。此外，对多种抗氧化酶和Ⅱ相代谢酶活性进行检测，其中绿茶组ＳＯＤ、ＧＳＴ、ＱＲ活性明显高于阳性对照组；混合茶组ＣＡＴ、ＳＯＤ、ＧＳＴ、ＱＲ活性明显高于阳性对照组。结果表明，绿茶和混合茶对ＤＥＮ诱发大鼠肝癌的发生均有显著的预防作用。其作用机理可能和诱导抗氧化酶及某些Ⅱ相代谢酶活性，以及抑制脂质过氧化作用有关。     

缺锌对大鼠脂质过氧化及抗氧化系统的影响

研究缺锌对３周龄大鼠的脂质过氧化和抗氧化系统的影响。方法：健康Ｗｉｓｔａｒ大鼠３３只,雌雄各半,按体重随机分为Ａ（缺锌组）,Ｂ（自由进食对照组）,Ｃ（配对喂养组）。每组１１只,实验期４０天。观察指标有血清和肝脏锌,肝脏超氧化物歧化酶（ＳＯＤ）、谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）活性,血清和肝脏丙二醛（ＭＤＡ）,并采用电子自旋共振（ｅｌｅｃｔｒｏｎｓｐｉｎｒｅｓｏｎａｎｃｅ,ＥＳＲ）及自旋捕捉技术直接测定大鼠肝组织中的氧自由基。结果：Ａ组大鼠肝脏ＳＯＤ、ＧＳＨ－Ｐｘ活性明显下降,血清和肝脏ＭＤＡ含量显著升高,ＥＳＲ图谱出现了Ｎ－Ｈ偶合的三组双峰波。结论：缺锌可以使大鼠脂质过氧化反应明显加强,而抗氧化能力明显减弱。     

铅对小鼠睾丸脂质过氧化作用的研究

为探讨铅的生殖毒性，研究了小鼠睾丸的脂质过氧化作用。将动物分为３组，对照组（Ⅰ），（１５ｍｇ／ｋｇ）（Ⅱ）和３０ｍｇ／ｋｇ（Ⅲ）铅染毒组，腹腔注射染毒１１次。结果表明，染毒组体重明显降低，血铅浓度比对照组增加１５０～２１０倍，骨铅增加７７～１０倍。Ⅰ、Ⅱ、Ⅲ组的睾丸ＴＳＯＤ分别为４０、３２、２４Ｎｕ／ｍｌ，差异具有显著性。血铅与睾丸ＴＳＯＤ有显著的负相关。睾丸ＧＳＨｐｘ也有明显改变。血浆维生素Ｃ和睾丸ＭＤＡ随着铅染毒剂量增加而有降低倾向。因此，脂质过氧化是铅导致生殖腺损伤的机理之一。     

Hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) combine CpG oligodeoxynucletides as a novel therapeutic vaccine for chronic hepatitis B infection

Hepatitis B virus infection is a non-cytopathic hepatotropic virus which can lead to chronic liver disease and hepatocellular carcinoma. Traditional therapies fail to provide sustained control of viral replication and liver damage in most patients. As an alternative strategy, immunotherapeutic approaches have shown promising efficacy in the treatment of chronic hepatitis B patients. Here, we investigated the efficacy of a novel therapeutic vaccine formulation consisting of two HBV antigens, HBsAg and HBcAg, and CpG adjuvant. This vaccine formulation elicits forceful humoral responses directed against HBsAg/HBcAg, and promotes a Th1/Th2 balance response against HBsAg and a Th1-biased response against HBcAg in both C57BL/6 and HBV transgenic mice. Vigorous cellular immune response was also detected in HBV transgenic mice, for a significantly higher number of HBs/HBc-specific IFN-γ secreting CD4+ and CD8+ T cells was generated. Moreover, vaccinated mice elicited significantly intense in vivo CTL attack, reduced serum HBsAg level without causing liver damage in HBV transgenic mice. In summary, this study demonstrates a novel therapeutic vaccine with the potential to elicit vigorous humoral and cellular response, overcoming tolerance in HBV transgenic mice.     

Preventing hepatitis B in people in close contact with hepatocellular carcinoma patients.

OBJECTIVE: To determine the prevalence of testing for hepatitis B virus (HBV) infection in the clinical management of primary liver cancer (hepatocellular carcinoma). METHODS: The authors reviewed the records of 78 patients treated for hepatocellular carcinoma in hospitals in the Puget Sound area in 1988 and early 1989 and reviewed all 1990 U.S. death certificates on which primary liver cancer was listed. RESULTS: The records of 50 (64%) of 78 hepatocellular carcinoma patients contained no evidence that the patient''s hepatitis B surface antigen (HBsAg) status had been determined. In addition, of 4353 people who died in 1990 for whom the diagnosis of primary liver cancer was listed on the death certificate, HBV infection was also listed for only 136 (3%), much less than expected based on case series. CONCLUSIONS: Many patients with hepatocellular carcinoma are not tested for HBV infection, suggesting that their close contacts are also not evaluated for HBV infection and the need for vaccination. Hepatitis B vaccination of close personal contacts of HBV-infected hepatocellular carcinoma patients is an important strategy for preventing HBV transmission.     

2008-2012年济南铁路单位职工乙肝病毒感染者肝癌发病情况观察分析

目的及时发现乙肝病毒（HBV）感染者中的肝癌患者,探讨HBV感染者肝功能与肝癌发病的关系。方法对2008年兖州6个铁路单位20～60岁健康体检职工中检出的HBV感染者485名,每半年进行一次肝功能、甲胎球蛋白（AFP）检测和肝脏B超检查随访,观察肝癌发病情况。结果2008～2012年12月31日,485名HBV感染者中,18人经AFP和（或）肝脏B超检查确诊为肝癌,肝癌累积发病率为3．71％。肝癌累积发病率,肝功正常者420人为1．43％,肝功异常者65人为18．46%（P〈0．01）;20～40岁225人为1．78％,41～60岁260人为5．38％（P〈0．05）。2008-2012年（观察第一年、第二年、第三年、第四年、第五年内）,485名HBV感染者肝癌发病率分别为0．62％、0．21％、0．62％、1．03％、1．24％。结论HBV感染者中肝功异常者容易发生肝癌,高年龄组尤为明显。     

Hepatitis B virus (HBV) markers among patients with chronic liver disease in Kuwait

The prevalence of hepatitis B surface antigen (HBsAg), antibody to the hepatitis B core (anti-HBc) and to surface antigen (anti-HBs), was investigated, using sensitive radioimmunoassay (RIA) systems, among patients with different clinical entities of chronic liver disease in Kuwait, and compared to a control blood donor population. 81% of patients and 44% of the controls had at least one HBV marker. 24% of patients, but none of the controls had both HBsAg and a high titre of anti-HBc in the absence of anti-HBs, suggesting a chronic infection. 31% of our patients with hepatosplenic schistosomiasis, 20% with cryptogenic cirrhosis and chronic active liver disease and 60% with hepatocellular carcinoma had these two markers. HBV antigenaemia was significantly more prevalent among male than among female patients and was particularly high among those less than 35 years old. The high prevalence of the various HBV markers among our patients suggests that HBV is a major factor in the development of chronic liver disease in our area. Furthermore, in view of the high prevalence of antigenaemia in patients with hepatosplenic schistosomiasis, HBV infection must play a concomitant role in the development of a more serious form of chronic liver disease among such patients.     

CMV-pp65抗原检测在乙肝相关性肝癌患者中的临床意义

目的:调查乙肝相关性肝癌患者CMV-pp65抗原血症情况,探讨CMV-pp65抗原检测的临床意义。方法:采用免疫组化二步法和酶联免疫吸附反应法(ELISA)检测30例乙肝相关性肝癌患者CMV-pp65抗原和CMV特异性IgM抗体;同时检测18例健康体检者的CMV-pp65抗原设为对照组,采用卡方检验进行统计分析。结果:30例乙肝相关性肝癌患者中CMV-pp65抗原血症阳性13例,阳性率43.3%(13/30);CMV特异性IgM抗体阳性2例,阳性率6.7%(2/30),经卡方检验,χ2=10.756,P<0.01,对比组之间的差异具有统计学意义。对照组18例健康体检者CMV-pp65抗原血症阳性0例,经卡方检验,χ2=10.697,P<0.01,对比组之间的差异具有统计学意义。结论:乙肝相关性肝癌患者CMV-pp65抗原血症阳性率高,提示活动性CMV感染;CMV-IgM抗体检测不适合肝癌患者活动性CMV感染的早期诊断。     

Does aflatoxin B1 play a role in the etiology of hepatocellular carcinoma in the United States?

Previous research showed that risk factors associated with hepatocellular carcinoma (HCC) include infection with hepatitis B (HBV) and hepatitis C (HCV) viruses, exposure to aflatoxin B1 (AFB1), and liver cirrhosis, due primarily to alcohol consumption. To determine whether AFB1 may play a role in HCC in the United States, a search for AFB1 adducts and p53 alterations, potentially induced by AFB1, was conducted in the United States in 23 HCC patients with available tissue samples. The presence of AFB1 tumor-DNA and -serum lysine adducts and mutant p53 product was determined by immunoassays and codon 249 p53 mutation by restriction enzyme analysis. HBV and HCV serology and serum HBV-DNA were also determined. Thirteen patients were positive for HBV by HBs antigen or anti-HBc antigen or by polymerase chain reaction for HBV-DNA sequences. Nine patients were free of HBV and HCV markers; 5 of 22 sera tested were anti-HCV positive. p53 Protein expression, determined by immunohistochemical staining, was present in 5 of the 23 tumor tissues, whereas p53 codon 249 mutations were not observed in the 5 cases in which tissue was available for study. AFB1 tumor-DNA adducts were present in 3 of 19 tumor tissues, and in 1 of these 3 samples p53 protein was also detected. Sera from only 5 of the patients were tested for AFB1-lysine adducts, and all were positive. In these five patients, neither p53 protein nor a mutation on codon 249 was detected. The demonstration that AFB1-DNA and -lysine adducts are present in HCC patients in the United States is intriguing but requires further substantiation because of the small number of subjects in this pilot study. To elucidate the pathogenetic significance of these findings, further investigation, including studies in larger patient cohorts and properly selected controls, is warranted.     

Inhibition of hepatocarcinogenesis in mice by dietary methyl donors methionine and choline

A dietary deficiency of methyl donors, the lipotropes methionine and choline, enhances the activity of hepatocarcinogens in rodents. To determine if the reverse is true, an excess of dietary choline, methionine, or both was fed to male mice given a carcinogenic dose of aflatoxin B1 (AFB1). Fifty weeks following the last dose of AFB1, all survivors were killed then examined for tumor incidence, and samples of nontumorous liver tissue were assayed for activities of mixed function oxidases (MFO). Survival was best in the high-methionine/high-choline group, with 36/38 surviving to termination of the study. Survival in the other groups was 35/38, 30/70, 33/38, and 34/37 in control with no AFB1, control with AFB1, groups with high methionine, and high choline, respectively. Combined adenoma/carcinoma incidence was 8/38, 30/37, 21/38, 20/37, and 10/38 in groups control with no AFB1, control with AFB1, high methione with AFB1, high choline with AFB1, and high choline and high methionine with AFB1, respectively. Cytochrome P450, cytochrome B5, cytochrome C, and ethylmorphine N-demethylase activities were all increased over controls, with most marked increases in the cytochrome P450 and ethylmorphine N-demethylase activities. The data presented here document a protective effect of dietary methyl donors on AFB1-induced hepatocarcinogenesis in mice probably acting, in part, via activation/detoxification mechanisms favoring an increased balance in detoxification of AFB1.     

Health Care Access and Sociodemographic Factors Associated with Hepatitis B Testing in Vietnamese American Men

Chronic hepatitis B viral (HBV) infection greatly increases the risk for cirrhosis and hepatocellular carcinoma. HBV serologic testing is important for the identification of chronically infected individuals, who may benefit from antiviral treatment and regular monitoring for disease sequelae. Elevated rates of cirrhosis and hepatocellular carcinoma among Vietnamese American men can largely be attributed to high rates of chronic HBV infection. We surveyed 509 Vietnamese men aged 18–64 years in Seattle, Washington and examined sociodemographic and health care access factors associated with HBV serology testing. Nearly two-thirds (65%) reported past testing. The following were among those factors associated with HBV testing in bivariate comparisons: older age; short proportion of life in the US; low English fluency; private health insurance; identifying a regular source of medical care; reporting no long waits for medical appointments; and having access to interpreter services. The following were independently associated with HBV testing in multiple logistic regression analysis: older age; college education; low English fluency; private health insurance; having a regular medical provider; and reporting no long waits for medical appointments. Younger and less educated men, and those with difficulty accessing medical care may be at particular risk for never having had HBV testing. Programs to reduce HBV transmission and sequelae should make special effort to target these vulnerable Vietnamese Americans.     

HLA—DR基因及乙肝病毒基因型与肝细胞癌相关性的初步研究

[目的]探讨乙型肝炎病毒（HBV）感染人群HLA—DR基因、HBV基因型与肝细胞癌的相关性。[方法]2007～2008年,应用序列特异性引物-聚合酶链反应（SSP—PCR）方法与荧光PCR（探针）法,分别对40例慢性乙肝患者（试验1组）、38例肝硬化患者（JR验2组）和34例肝细胞癌患者（试验3组）的HLA—DR等位基因、HBV基因型进行检测,并以中华骨髓库山东分库无血缘关系的自愿骨髓捐献者1383例为对照,分析其表达与肝细胞癌的相关性。[结果]HLA—DR1基因的检出率,试验3组为14．71％,对照组为4．85％（P〈0．05）;HLA—DR13基因的检出率,试验3组为14．71％,试验1组为0．00％（P〈O．05）;试验2组与试验3组各等位基因表型的检出率差异均无统计学意义（P〉0．05）。HBV基因型B型、C型、非B非C型所占比例,试验1、2、3组的差异无统计学意义（P〉0．05）。试验3组中,HLA—DR1、DR13阳性的5例均为HBV基因C型;其他29例HBV基因C型23例、非B非C型6例（P〉0．05）。[结论]HLA—DR1可能是肝细胞癌的易感基因．携带HLA—DR13基因位点的HBV感染者更易发展为肝细胞癌。     

Lentivector expressing HBsAg and immunoglobulin Fc fusion antigen induces potent immune responses and results in seroconversion in HBsAg transgenic mice

Even though hepatitis B virus(HBV) vaccines effectively prevent new cases of HBV infection, with approximately 350 million patients worldwide, chronic HBV infection remains a major health problem because of the associated complications (such as liver cirrhosis and hepatocellular carcinoma) and the limited treatment options. Immunotherapy has the potential to effectively control HBV replication. In this current study, we found that recombinant lentivectors could induce potent HBV surface antigen (HBsAg) specific T cell responses and humoral immune responses. Tagging the HBsAg with immunoglobulin Fc fragment further substantially increased the HBsAg specific immune responses. Remarkably, the HBS-Fc-lv lentivector could effectively break immune tolerance and induce potent HBsAg specific adaptive immune responses in HBsAg transgenic (Tg) mice with low serum level of HBsAg. More importantly, the induction of HBsAg specific immune responses in Tg mice accompanied seroconversion from HBsAg to anti-HBsAg antibody (anti-HBsAb). Our study demonstrated the potential of utilizing lentivector to treat chronic HBV infection following reduction of viral load with antiviral drug therapy.