牛带绦虫乳酸脱氢酶基因原核表达及免疫学特征

目的 克隆牛带绦虫乳酸脱氢酶(lactate dehydrogenase,LDH)基因在大肠杆菌中表达,研究其免疫学特征。方法 将牛带绦虫成虫LDH基因与质粒pET-28a(+)连接构建原核重组质粒,在大肠埃希菌BL21/DE3中用异丙硫代-β-D半乳糖苷诱导表达,表达产物通过十二烷基-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白印迹(Westernblotting)进行分析。结果 牛带绦虫乳酸脱氢酶基因编码332个氨基酸,理论分子量为35.4kDa,PCR、双酶切及DNA测序的结果均证明pET-28a(+)-LDH构建成功;SDS-PAGE结果表明,牛带LDH基因在大肠埃希菌高效表达,经过亲和层析高纯度蛋白,且该重组蛋白可以被亚洲带绦虫、牛带绦虫及猪带绦虫感染病人血清识别,表明其具有免疫反应性。结论 牛带绦虫LDH基因可在原核系统中有免疫活性的高效表达。     

肠道寄生虫病药物治疗进展

在人体寄生虫病的范畴中 ,肠道寄生虫病占有极大的比重和数量。根据 1988～ 1991年全国人体寄生虫分布调查 ,肠道线虫感染率为 5 9.2 % ,估计全国蛔虫感染的人数为 5 .31亿人 ,钩虫感染为 1.94亿人 ,鞭虫感染为 2 .12亿人 ,而且有相当一部分人群混合感染 2种以上的肠道寄生虫[1] 。因此我国防治肠道寄生虫病的任务已列入卫生部十·五寄生虫病的防治规划。在防治寄生虫病的措施中 ,实践证明化学药物治疗是非常重要的一环 ,不仅对恢复病人健康、保护劳动力 ,而且对控制或消灭传染源都有很重要的现实意义。现将有关常用药物及近年开发的新化学…     

阿苯达唑和甲苯咪唑对感染旋毛虫沙鼠的影响

旋毛虫病是旋毛虫 (Trichinellaspiralis)成虫和幼虫分别寄生于同一宿主小肠和肌细胞内而引起的人兽共患寄生虫病。阿苯达唑 (Albendazole)和甲苯咪唑 (Mebendazole)是广谱、高效的抗蠕虫药 ,对多种寄生蠕虫及某些组织内寄生幼虫有良好的杀灭作用 ,2种药物对旋毛虫成虫和幼虫均有杀伤力大、疗效高 ,其中前者已列为治疗旋毛虫病的首选药物〔1,2〕。本文进一步观察阿苯达唑和甲苯咪唑对感染旋毛虫的沙鼠在幼虫移行期和囊包形成期血清中ALT、AST、ALP、CK酶的影响。材料与方法　 (1)实验动物 :8周龄长爪沙鼠 ,体重 45～60 g ,雌雄各半 ,…     

苯系混合物对男工精液乳酸脱氢酶同工酶的影响

苯系混合物对男工精液乳酸脱氢酶同工酶的影响肖国兵潘翠宝周承来任世女我们开展了苯系物对作业男工精液乳酸脱氢酶同工酶Ｃ４（ＬＤＨ－Ｃ４）活性影响的研究。一、对象与方法１．调查对象：选择油（浸）漆、造漆和制鞋等行业接触苯系物１年以上，无慢性疾病史，尤其无生...     

脑卒中对血糖、异柠檬酸脱氢酶、乳酸脱氢酶的影响

目的 研究脑卒中患者血糖（BS）、异柠檬酸脱氢酶（IDH）、乳酸脱氢酶（LDH）水平的变化。方法采用葡萄糖氧化酶法检测67例非糖尿病性脑卒中患者的血糖水平,根据血糖值分为高血糖组（41例）和正常血糖组（26例）,另设24例体检健康者为对照组,用比色法检测血清IDH、LDH水平,用琼脂糖凝胶电泳法测定乳酸脱氢酶同工酶（LDH-同工酶）。结果与对照组及正常血糖组相比,高血糖组患者血清IDH、LDH水平显著增高,P〈0．01：对照组与正常血糖组相比较,差异无显著性。结论脑卒中可影响患者BS、血清IDH及LDH水平,非糖尿病脑卒中患者的BS、IDH、LDH水平高,病情较重。     

日本血吸虫乳酸脱氢酶原核表达、纯化及鉴定

目的原核表达日本血吸虫乳酸脱氢酶（SjLDH）,对表达产物进行纯化及生物活性鉴定。方法将SjLDH编码序列克隆入pET-28a原核表达载体并转染大肠埃希菌,异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达后亲和层析法对表达产物进行纯化,并用聚丙烯酰胺变性凝胶电泳（SDS-PAGE）、蛋白印迹（Western blot）、酶联免疫吸附实验（ELISA）、酶活性染色及活性测定进行鉴定。结果成功构建pET28a-SjLDH重组质粒并在大肠埃希菌中获得可溶性表达,亲和层析纯化表达产物。SDS-PAGE及Western Blot结果显示,表达及纯化产物分子量约36kDa,与SjLDH理论分子量相符。ELISA结果表明,重组蛋白具有良好的免疫活性,酶活性染色表明重组蛋白具有乳酸脱氢酶活性,活性单位为379U/mg。结论成功进行SjLDH的原核表达并获得纯化的重组蛋白,重组蛋白具有良好的免疫原性及酶活性,为研究SjLDH的功能奠定了基础。     

200例脑囊虫病疗效及不良反应分析

目的 评价阿苯达唑、吡喹酮治疗脑囊虫病的疗效及不良反应.方法 将2002-2007年收治的脑囊虫病病例资料,随机抽取200例,分单纯使用阿苯达唑治疗组58例和吡喹酮、阿苯达唑联合治疗组142例两组,进行上述目的 分析.结果 治疗后两组临床有效率差异无统计学意义(P>0.05);颅内高压症、癫痫发作、精神症状的不良反应发生率阿苯达唑组少于联合治疗组,差异有统计学意义(P<0.05);过敏反应两组差异无统计学意义(P>0.05).结论 阿苯达唑、吡喹酮治疗脑囊虫病疗效肯定,但吡喹酮和阿苯达唑联合用药的不良反应较阿苯达唑单独用药严重.     

过氧化苯甲酰对大鼠睾酮和乳酸脱氢酶的影响

[目的]观察过氧化苯甲酰对雄性大鼠血清睾酮和乳酸脱氢酶含量的影响。[方法]将40只雄性大鼠随机分为4组,其中1组为对照组,其余3组分别为过氧化苯甲酰低、中、高剂量组。16周后检测血清睾酮和乳酸脱氢酶的含量。[结果]对照组、低剂量组、中剂量组、高剂量组大鼠体重比较,差异有统计学意义(F=6.847,P﹤0.01),对照组、低剂量组、中剂量组、高剂量组大鼠血清睾酮含量比较差异有统计学意义(F=8.874,P﹤0.01),对照组、低剂量组、中剂量组、高剂量组大鼠血清乳酸脱氢酶含量比较,差异有统计学意义(F=6.378,P﹤0.01)。[结论]过氧化苯甲酰能降低大鼠体重和血清睾酮、乳酸脱氢酶的含量。     

亚洲牛带绦虫TaCRISP基因克隆、表达和序列分析

目的通过筛选亚洲牛带绦虫(Taenia saginata asiatica)成虫cDNA质粒文库,识别半胱氨酸分泌蛋白(Ta CRISP),并进行克隆表达,为进一步研究其功能提供基础依据。方法用生物信息学方法从亚洲牛带绦虫成虫全长cDNA质粒文库中识别TaCRISP的全长编码基因并预测编码蛋白质的各种结构与功能;将其编码区序列克隆到原核表达载体pET-30a(+)上,测序鉴定重组质粒,之后进行诱导表达,表达产物经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定。结果该基因全长1090bp,编码239个氨基酸,1aa-16aa位为分泌信号肽,理论分子量为26973.8,等电点为5.96。生物信息分析揭示,Ta CRISP与中殖孔绦虫CRISP一致性为38%,相似性为54%;所构建的重组体经PCR、双酶切及测序鉴定与目标基因相符。SDS-PAGE结果表明,目的基因在大肠埃希菌BL-21/DE3中表达成功。结论发现亚洲牛带绦虫Ta CRISP基因,成功构建重组原核表达质粒并表达出融合蛋白。     

转运蛋白和乳酸脱氢酶在乳腺癌糖酵解中的作用

有氧糖酵解途径是肿瘤细胞特征性的代谢方式.多数肿瘤细胞通过提高葡萄糖摄取率、增强糖酵解活性和产生大量乳酸,来促进肿瘤细胞的生长和转移,包括乳腺癌.乳腺癌是女性最常见的肿瘤,也是最主要的肿瘤性死亡原因.乳腺癌大多具有糖酵解活性,其中葡萄糖转运蛋白(GLUT)、单羧酸转运蛋白(MCT)和乳酸脱氢酶(LDH)等糖酵解关键酶表...     

亚洲牛带绦虫WD40基因表达及免疫学分析

目的 对亚洲牛带绦虫WD40基因进行克隆、表达和免疫学研究.方法 将亚洲牛带绦虫成虫WD40基因克隆到原核表达质粒pET-28a( )中,在大肠埃希菌BL-21/DE3中用异丙硫代-β-D半乳糖苷(IPTG)诱导表达,表达产物通过十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)进行鉴定,用镍离子金属螯合剂亲和层析柱进行纯化,纯化的重组蛋白用免疫印迹试验(Western Blotting)进行免疫学分析.结果 PCR、双酶切及DNA测序结果均表明,pET-28a( )-WD40重组质粒构建成功.SDS-PAGE结果表明,目的 基因在大肠埃希菌BL-21/DE3中获得高效表达.重组蛋白能识别感染了亚洲牛带绦虫的猪血清和患者血清,表明其具有免疫反应性.结论 亚洲牛带绦虫WD40基因可在原核表达系统中获得具有免疫学活性的表达,为进一步研究该蛋白的功能提供了基础依据.     

亚洲带绦虫囊尾蚴特异性抗原筛选和鉴定

目的 对亚洲带绦虫囊尾蚴的特异性抗原进行筛选、鉴定和生物信息学分析.方法 6头20d龄三元杂交乳猪随机分为实验组和对照组,实验组每头灌胃亚洲带绦虫虫卵悬液8.4万个;20 d后,分别制备实验组及对照组心脏血混合血清;收集、制备实验组亚洲带绦虫囊尾蚴蛋白并进行双向电泳分析,将凝胶蛋白斑点转移至聚偏氟乙烯膜,用实验组及对照组混合血清(均为1∶10)为一抗,羊抗猪辣根过氧化物酶- IgG(1∶200)为二抗,进行蛋白质印迹分析和质谱鉴定.结果 双向电泳凝胶共检测到( 204±11)个蛋白质斑点,相对分子质量为14 400 ～94 000 kPa,等电点(pI)为3.0～10.0;筛选出6个特异性抗原,经质谱鉴定出其中2个蛋白,分别为ATP酶管家基因3-类蛋白2(AT-Pase family gene 3-like protein 2)和ATP合成酶β亚基(ATP synthase beta subunit),均与美国国家生物技术信息中心(NCBI)网上登录的亚洲带绦虫成虫蛋白一致,确定为亚洲带绦虫囊尾蚴特异性抗原.结论 获得2个亚洲带绦虫囊尾蚴特异性抗原,其免疫原性有待进一步研究,以便为研制抗囊尾蚴病疫苗提供参考.     

脑脊液乳酸脱氢酶检查在小儿颅内感染中的意义

目的 研究和分析脑脊液乳酸脱氢酶检查对小儿颅内感染中鉴定小儿化脓性脑膜炎以及病毒性脑膜炎的意义.方法 分析201 3年1-6月进入该院治疗化脓性脑膜炎患儿52例,病毒性脑膜炎患儿48例,以及对照组患儿50例,检测脑脊液乳酸脱氢酶的浓度.结果 脑脊液乳酸脱氢酶浓度在病毒性脑炎组（24.5±3.7）U/L,对照组病例（2.2±3.56）U/L,化脓性脑膜炎组结果（54.6±3.4）U/L与其他两组检测结果相比显著增高（P＜0.05）.结论 脑脊液乳酸脱氢酶检测能够作为鉴别小儿化脓性脑膜炎和病毒性脑炎更为快速和准确的诊断标准,对临床上小儿颅内感染疾病的诊断治疗提供更为准确的实验室数据指标,有助于提高小儿颅内疾病的治疗效率.     

Feasibility of baculovirus-expressed recombinant 10-kDa antigen in the serodiagnosis of Taenia solium neurocysticercosis

Bacterially expressed recombinant 10-kDa protein of Taenia solium metacestode (TsM) was previously found to be reliable in the diagnosis of active stage neurocysticercosis (NCC) by immunoblotting but not by ELISA. In this study, we evaluated the diagnostic feasibility of detecting eukaryote-expressed recombinant 10-kDa protein of TsM by ELISA (rTsM10-ELISA) in the serum and cerebrospinal fluid (CSF) from NCC patients. In 45 cases of active NCC, 91.1 and 97.8% cases showed positive reactions for serum and CSF by rTsM10-ELISA. ELISA employing the crude cyst fluid antigen (CF-ELISA) also revealed a similar result. Negligible cross-reactions were observed in serum samples from control subjects and from subjects with other helminthic diseases by rTsM10-ELISA (5/139 cases, 3.6%). By contrast, CF-ELISA demonstrated a high degree of cross-reactivity (24/139, 17.3%) especially from those patients with alveolar and cystic echinococcoses. The overall sensitivity and specificity of rTsM10-ELISA were 94.3 and 96.4%; and those of CF-ELISA were 95.7 and 84.5%, for serum and CSF, respectively. Antibody responses to rTsM10 were detected as early as 3 months after experimental infection of T. solium eggs in pigs. Our results show that ELISA with rTsM10 could be highly applicable in the serodiagnosis of NCC from early stage of infection.     

乳酸脱氢酶同功酶活性测定应用研究

我国卫生部１９８５年已将小鼠精子畸形试验作为可选择的致突变试验方法，但是精子发生是一个多时相的复杂过程，采样时错过了精子发育的敏感阶段，或因感染，天气炎热体温变化，致使实验得不到预期结果。为了探讨血浆中ＬＤＨ－Ｘ酶活性测定与精子畸形率的关系设计了本实验研究。三种阳性物（ＭＭＣ、ＣＰ、Ｃｄ）均对睾丸细胞产生明显的损伤作用，睾丸匀浆及血浆中ＬＤＨ－Ｘ酶活性、精子畸形率均显著增高，表明三者结果基本一致。本文还对血浆中ＬＤＨ－Ｘ酶活性测定的应用进行了讨论。     

Evaluation of a new Plasmodium lactate dehydrogenase assay (OptiMAL-IT) for the detection of malaria

The new OptiMAL-IT(R) rapid diagnostic test for malaria was evaluated in 271 patients in Thailand with uncomplicated malaria between June and July 2002. The sensitivity and specificity for the diagnosis of Plasmodium falciparum parasites were 88% and 92%, respectively. For species other than P. falciparum, the sensitivity was 65% and specificity was 99%. The performance of the new test decreased markedly at low levels of parasitaemia.     

乳酸脱氢酶和降钙素原及C-反应蛋白联合检测在小儿中枢神经系统感染诊断中的意义

目的:探讨乳酸脱氢酶(LDH)、降钙素原(PCT)及C-反应蛋白(CRP)联合检测在中枢神经系统感染(CNSI)患儿早期诊断中的临床价值,为CNSI患儿的治疗提供参考依据.方法:选取2011年10月～2014年4月聊城市第二人民医院收治的80例CNSI患儿为病例组,包括40例病毒性感染患儿和40例细菌性感染患儿;同时随机抽取40例无CNSI的儿童为对照组.对病例组和对照组患儿均进行LDH、PCT和CRP联合检测,分析其诊断价值.结果:细菌性感染组的LDH、PCT和CRP水平显著高于病毒性感染组和对照组,差异有统计学意义(P＜0.05);病毒性感染组的LDH和CRP水平显著高于对照组,差异有统计学意义(P＜0.05),而PCT与对照组比较差异无统计学意义(P＞0.05).结论:LDH、PCT和CRP联合检测对CNSI患儿的诊断有较高的价值,可鉴别细菌性和病毒性感染.     

The heat stability of Plasmodium lactate dehydrogenase-based and histidine-rich protein 2-based malaria rapid diagnostic tests

Malaria rapid diagnostic tests (RDTs) have performed well in a variety of studies, but recent reports have described sensitivity for Plasmodium falciparum as significantly lower than that required for operational deployment. Exposure to high temperature has been suggested as an explanation. This study assessed the temperature stability of two different Plasmodium lactate dehydrogenase (pLDH)- and three histidine-rich protein 2 (HRP2)-detecting RDTs. One HRP2 test proved insufficiently sensitive for assessment. After incubation at 35, 45 and 60 degrees C, two RDTs detecting pLDH showed a substantial fall in percentage test line positivity over time, which was not seen with the remaining two HRP-2-based RDTs. For the particular products studied, variability was high, with the pLDH-based RDTs being less sensitive than HRP2-based RDTs against the sample of P. falciparum used and more susceptible to heat-induced damage, but the reasons for this are unclear. The performance of malaria RDTs can be adversely affected at the temperatures to which they will be exposed when transported to, and used in, the rural tropics.     

Assessments of pharmacokinetic drug interactions and tolerability of albendazole, praziquantel and ivermectin combinations

The pharmacokinetic interactions and tolerability of albendazole, praziquantel and ivermectin combinations were assessed in 23 healthy Thai volunteers (12 males and 11 females). The study was an open, randomised, three-way crossover design in which each subject attended the study on three separate occasions (Phases I, II and III), of 4 d or 8 d each, with at least 1 or 2 weeks (but not longer than 2 months) between each phase. All subjects received the three study drug regimens as follows: regimen I, oral praziquantel (40 mg/kg body weight); regimen II, oral ivermectin (200 microg/kg body weight) given concurrently with an oral dose of albendazole (400 mg); and regimen III, oral ivermectin given concurrently with albendazole and praziquantel. All treatment regimens showed acceptable tolerability profiles. The incidence of overall drug-related adverse events was significantly higher following regimens I (12/23) and III (7/23) compared with that following regimen II (0/23). Six statistically significant changes in the pharmacokinetic parameters of albendazole sulphoxide (Cmax, AUC0-infinity, Vz/F, CL/F), praziquantel (Vz/F) and ivermectin (AUC0-infinity) were observed when the three drugs were given concurrently. However, based on US Food and Drug Administration criteria, these changes were not considered of clinical relevance.