Effect of Oenanthe Javanica Extract on Antioxidant Enzyme in the Rat Liver

Background:

Oenanthe javanica (O. javanica) has been known to have high antioxidant properties via scavenging reactive oxygen species. We examined the effect of O. javanica extract (OJE) on antioxidant enzymes in the rat liver.

Methods:

We examined the effect of the OJE on copper, zinc-superoxide dismutase (SOD1), manganese superoxide dismutase (SOD2), catalase (CAT), and glutathione peroxidase (GPx) in the rat liver using immunohistochemistry and western blot analysis. Sprague-Dawley rats were randomly assigned to three groups; (1) normal diet fed group (normal-group), (2) diet containing ascorbic acid (AA)-fed group (AA-group) as a positive control, (3) diet containing OJE-fed group (OJE-group).

Results:

In this study, no histopathological finding in the rat liver was found in all the experimental groups. Numbers of SOD1, SOD2, CAT, and GPx immunoreactive cells and their protein levels were significantly increased in the AA-fed group compared with those in the normal-group. On the other hand, in the OJE-group, numbers of SOD1, SOD2, CAT, and GPx immunoreactive cells in the liver were significantly increased by about 190%, 478%, 685%, and 346%, respectively, compared with those in the AA-group. In addition, protein levels of SOD1, SOD2, CAT, and GPx in the OJE-group were also significantly much higher than those in the AA-group.

Conclusion:

OJE significantly increased expressions of SOD1 and SOD2, CAT, and GPx in the liver cells of the rat, and these suggests that significant enhancements of endogenous enzymatic antioxidants by OJE might be a legitimate strategy for decreasing oxidative stresses in the liver.     

Assessing donor suitability for blood donation: Utility of Geenius HIV 1/2 confirmatory assay

BackgroundBlood donor care and blood safety require a quick and accurate decision on the presence or absence of Human Immunodeficiency Virus (HIV) infection, based on the proper selection of blood donors, serological and molecular HIV testing as well as western blot test. The aim was investigating the possibility of inclusion of Geenius HIV 1/2 Confirmatory Assay in blood donor testing algorithm in order to shorten test time and decrease the number of indeterminate results.MethodsA total of 75 archived serum/plasma samples were tested. Their previous serological and molecular HIV results were: 3 negative samples, 7 positive samples, 65 serological indeterminate or positive but confirmatory testing and NAT negative samples.ResultsGeenius assay confirmed the presence of antibodies in all blood donors with HIV positive serology and Nucleic Acid Testing (NAT). HIV-1 gp160 and gp41 antibodies were detected in these donors, while p31 and p24 antibodies were not detected in two and three donors, respectively. HIV-2 antibodies gp36 and gp140 were not found. Blood donor with HIV indeterminate or positive serology but negative confirmatory testing and NAT, were negative in Geenius assay.Conclusion The results obtained are consistent with western blot results. The assay proved simple and quick to perform. Studies have confirmed the possibility of introducing Bio-Rad Geenius into a routine blood donor testing protocol.     

杂色曲霉素对人外周血单个核细胞表面HLA-Ⅰ分子表达的影响

目的探讨杂色曲霉素(ST)对体外培养的人外周血单个核细胞表面(HPBMc)HLAⅠ分子表达的影响。方法采用流式细胞定量术(FCM)和免疫印迹(Westernblot)分析方法,研究不同浓度ST(0.125、0.25、0.5、1和2mgL)处理后人外周血单个核细胞表面HLAⅠ分子表达的变化。结果FCM定量分析结果表明,经不同浓度ST处理24小时后,与对照组相比,各组细胞HLAⅠ的平均荧光强度均降低,以较高浓度(0.5、1和2mgL)ST处理组降低更明显(P<0.05)。在0.125mgL到2mgL的浓度范围内,随ST处理浓度的升高,HLAⅠ荧光指数逐渐降低,两者呈明显的负相关(r=-0.841,P<0.01)。免疫印迹结果也表明,随ST浓度的增高,HLAⅠ分子降低越明显。结论提示在0.125mgL到2mgL的浓度范围内,ST抑制人外周血单个核细胞表面HLAⅠ分子的表达呈现出负的剂量-反应关系。     

用蛋白印迹技术初步评价肝母细胞瘤血管内皮生长因子表达的意义

目的通过观察小儿肝母细胞瘤（pedo-hepatoblastoma）组织中血管内皮生长因子C（vascular endothelial cell growth factor C，VEGF-C）及其受体Fh4（又称VEGF-R3）蛋白的表达，初步评价其生物学行为间的关系及检测的意义。方法用蛋白印迹技术检测26例小儿肝母细胞瘤、瘤旁肝组织及20例正常对照组织中蛋白的表达。结果19例肝母细胞瘤有明显的VEGF-C蛋白印迹（73．1％），7例瘤旁肝组织中有不同程度的VEGF-C蛋白印迹（26．9％）；21例肝母细胞瘤有明显的Fh4蛋白印迹（80．8％），6例瘤旁肝组织中有不同程度的Flt4蛋白印迹（23．1％）。20例正常对照组织中均不与VEGF-C和Fh4反应，而没有蛋白表达。结论VEGF-C和Fh4在肝母细胞瘤中呈高表达，可能在肝母细胞瘤分化、浸润及病理组织类型中起作用；其蛋白的检测可能有一定的临床意义。     

尖锐湿疣患者HPV DNA的检出及其与外周血淋巴细胞表型关系的研究

目的探讨尖锐湿疣(CA)患者人乳头瘤病毒(HPV)DNA的存在与细胞免疫应答的关系.方法应用核酸印迹技术(Southernblot)对30例CA患者的活体组织进行HPV的检测,并通过限制性片段长度多态性(RFLP)分析定型.应用抗CD3、CD4、CD8单克隆抗体致敏的红细胞花环试验法(SERC),同步检测外周血T细胞亚群.结果30名CA患者中,HPV阳性者19例(63.3%).其中,HPV11型15例(78.9%,15/19),HPV6型3例(15.8%,3/19),HPV16型1例(5.2%,1/19).CA病人外周血中CD3细胞、CD4细胞的百分比明显低于对照组(t值分别为3.32、3.42,P<0.01),CD8细胞的百分比明显高于对照组(t=12.42,P<0.001),CD4/CD8比值与对照组相比下降显著(t=10.67,P<0.001).结论CA患者体内由于HPVDNA的存在致使多种T细胞亚群功能障碍所引起的一系列细胞免疫应答抑制效应,在CA发病机理中起着重要作用.     

核因子-κB p65与心肌素在子宫肌瘤中的表达及作用机制

目的探讨核因子(NF)-κB p65亚型和心肌素在子宫肌瘤发生中的作用机制。方法蛋白印迹法检测29例子宫肌瘤和相应肌层组织以及15例原代子宫肌瘤细胞和相应肌层细胞中NF-κB p65、心肌素的表达;RNA干扰技术抑制子宫肌瘤细胞中NF-κB p65基因表达,光镜观察细胞生长状况,蛋白印迹法检测干扰后子宫肌瘤细胞中NF-κB p65和心肌素的表达。结果与正常肌层相比,子宫肌瘤组织和细胞中的NF-κB p65相对表达量增高(P<0.05),而心肌素的相对表达量降低(P<0.05)。RNA干扰后,与未转染的子宫肌瘤细胞相比,NF-κB p65的相对表达量降低(P<0.05),而心肌素的表达量相对增高(P<0.05)。结论 NF-κB p65与心肌素可能相互作用共同参与了子宫肌瘤的形成。     

Analytical technology development to monitor the stability of Polysaccharide-Protein conjugate vaccines

The covalent attachment of a bacterial-derived capsular polysaccharide to protein is of critical importance in transforming the polysaccharide from an antigen with limited immunogenicity in infants and older adults to an antigen that can prevent potentially fatal disease. For a polysaccharide-protein conjugate vaccine (PCV) candidate to be successful, it must be sufficiently stable. Chemical breakage of carbohydrate bonds in the polysaccharide may result in the reduction of “conjugate dose” and could negatively impact immunogenicity and the ability of the vaccine to prime for memory responses. Therefore, development of analytical tools to monitor the integrity of a polysaccharide-protein conjugate (glycoconjugate) vaccine is of practical significance. In this work, reducing SDS-PAGE, Intrinsic Protein Fluorescence Spectroscopy (IPFS), Differential Scanning Fluorimetry (DSF) were evaluated methods to study the impact of time, temperature, and formulation composition on the stability of a glycoconjugate vaccine prepared by multisite coupling of polysaccharide to a carrier protein. In addition, an automated capillary Western system was also evaluated to study the impact of storage on glycoconjugate vaccine stability. Two streptococcus pneumoniae polysaccharide-protein conjugates (serotype 3 and serotype 19A) were chosen to examine their physicochemical stability when formulated as a single antigen vaccine. While all methods require only a small amount of test article and can test multiple samples per assay run, automated capillary Western has the additional advantage of being highly sensitive even at low concentrations in complex vaccine formulations that contain aluminum adjuvant and multiple antigens. Results suggest that automated capillary Western is stability-indicating and may be an effective analytical technology tool for the formulation development of a multivalent glycoconjugate vaccine.     

慢性砷中毒对小鼠齿状回GFAP表达的影响

目的 研究砷中毒后小鼠齿状回胶质原纤维酸性蛋白(GFAP)的变化。方法 选取健康成年昆明小鼠80只,雌雄各半,分为对照组及慢性砷中毒高、中、低剂量组,每组20只,分别以蒸馏水、1/5 LD50、1/10 LD50、1/40 LD50 As2O3连续灌胃3个月,根据其体质量变化随时调整用药剂量,采用Y-电迷宫检测各组小鼠学习记忆行为,采用免疫组织化学和蛋白印迹技术检测不同浓度砷中毒对小鼠齿状回部位GFAP表达的影响。结果 与正常对照组比较,高剂量砷中毒组学习、记忆Y-迷宫测试次数明显增多(P<0.05),小鼠齿状回GFAP阳性细胞明显增多(P<0.01),阳性反应产物平均光密度值增高(P<0.01),随砷中毒剂量的增加,小鼠齿状回GFAP蛋白含量随之增高(P<0.01)。结论 慢性砷中毒引起的学习记忆损伤,可能与齿状回神经胶质细胞反应性增生及GFAP表达增强有关。     

博尔纳病病毒感染与神经胶质瘤相关性研究

目的探讨博尔纳病病毒(BDV)感染与神经胶质瘤发生的可能相关性。方法用Western-blot方法,对23例神经胶质瘤患者和23例脑外伤患者血清中BDV-p24/p40特异性抗体进行检测。结果 23例神经胶质瘤患者血清中BDV-p24/p40抗体阳性4例,阳性率为17.4%;脑外伤患者血清标本中无BDV-p40抗体检出,阳性率为0%。结论神经胶质瘤患者血清中存在博尔纳病病毒的感染。     

小鼠瘦素基因的原核表达及产物纯化

目的 应用大肠杆菌表达系统进行小鼠瘦素基因的克隆、表达、产物纯化与鉴定.方法应用聚合酶链扩增技术,以含有小鼠瘦素cDNA序列的质粒pMET mouse leptin为模板,扩增后克隆至载体pET-28a(+)构建重组表达载体pET-28a-lep,酶切、测序正确后,转化大肠杆菌E. coli BL21,构建工程菌株BL21-pET-lep.使用0.1mmol/L异丙基-硫代半乳糖苷,在不同的时间段诱导蛋白表达,利用十二烷基磺酸钠-聚丙烯酰胺电泳和Western Blotting检测瘦素蛋白的分子质量以及最佳表达时间.使用0.5%十二烷基肌氨酸钠溶解包涵体后通过Ni2+亲和层析柱纯化.结果 正确构建了表达载体pET-28a-lep.0.1 mmol/L异丙基硫代半乳糖苷诱导BL21-pET-lep 6 h具有最高蛋白表达量,含量占菌体总蛋白的39.2%.SDS-PAGE以及Western Blotting检测显示所表达的小鼠瘦素为携带组氨酸标签的融合蛋白,分子质量约为22.5 kDa.使用Ni2+层析柱纯化后的蛋白纯度为1.086 g/L.结论 应用大肠杆菌原核表达系统成功进行了小鼠瘦素基因的克隆、表达、产物纯化与鉴定.