槐果碱对感染CVB3搏动心肌细胞的保护作用

目的：应用柯萨奇病毒B3(CVB3)感染SD大鼠乳鼠心肌细胞造成病毒性心肌炎模型,观察槐果碱对该模型的作用。方法：(1)将乳鼠心肌细胞分成4组：感染组,感染CVB3；治疗组,感染病毒后加入100mg·L~(-1)的槐果碱；同时设立药物对照组和细胞对照组。在处理的d 2,3,5观察各组细胞病变效应(CPE)、细胞搏动频率并测定细胞上清液中乳酸脱氢酶浓度。(2)将心肌细胞感染CVB3后加入不同浓度的槐果碱(12．5～400mg·L~(-1)),设立病毒、细胞及药物对照。在处理后d 2,3,5观察CPE并测定细胞上清中的乳酸脱氢酶(LDH)浓度。结果：(1)100mg·L~(-1)的槐果碱对感染CVB3的乳鼠心肌细胞有保护作用,治疗组的CPE较感染组减轻,上清液中的LDH浓度降低。(2)槐果碱在12．5～300mg·L~(-1)之间均对感染CVB3的乳鼠心肌细胞有保护作用,它可以减轻病毒导致的CPE,并减少LDH释放。在400mg·L~(-1)反而会加重CPE,增加LDH释放。结论：一定浓度的槐果碱在体外对感染CVB3的心肌细胞有保护作用。     

丙型肝炎病毒在唾液腺中增殖

非甲非乙型肝炎可通过皮下接种唾液实验性感染黑猩猩。应用聚合酶链反应(PCR)可在血清丙型肝炎病毒(HCV)阳性病人的唾液中检出HCV。尽管唾液中HCV被认为源于唾液腺,但未被证实。由于HCV的基因组系正极单股RNA分子,在任何组织中负股HCVRNA的选择性检出证实HCV能在该组织中复制并感染该组织。本文报道在尸体解剖病     

地胆草提取物对肝癌血管生成拟态的作用研究

目的 研究地胆草提取物对肝癌HepG2细胞增殖、迁移、血管生成拟态的作用和机制.方法 取对数生长期HepG2肝癌细胞,分为对照组(0.1％DMSO)和不同浓度实验组(地胆草提取物质量浓度分别为25,50,75,100,125,150μg·mL-1).以噻唑蓝(MTT)比色法测定地胆草提取物对HepG2细胞增殖的影响;根...     

丙型肝炎病毒全长cDNA克隆在Huh7细胞中的表达与复制

目的 了解丙型肝炎病毒(HCV)在体外的复制和表达情况。方法 利用HCV全长cDNA克隆构建表达质粒p3．1HCV并转染Huh7细胞。应用RT—PCR，Western blotting分析病毒正、负链RNA，蛋白表达和剪切。结果 在转染p3．1HCV的Huh7细胞中可检出相应病毒的正、负链RNA、Western blotting在转染p3．1HCV的Huh7细胞检测中可检到针对HCVNS3蛋白、分子量约70kD的特异性条带。结论 HCV可以在体外培养的Huh7细胞中复制且其前体蛋白能在Huh7细胞中完成剪切。     

慢性丙肝患者干扰素治疗血清中丙肝病毒负链RNA的检测及临床意义

利用套式逆转录多聚酶链反应（RT－PCR）方法，对17例慢性丙肝患者进行了血清丙肝病毒正、负链RNA的检测，并且动态观察了其对干扰素治疗的反应。结果发现治疗前17例患者血清中正链RNA均阳性，13例血清负链RNA亦阳性（反应组8例，无反应组5例）。治疗后反应组中4例血清正链RNA阴转，3例负链RNA阴转；无反应组中血清正、负链RNA状况无变化。提示患者血清中有正、负链RNA复制复合体存在，体内有活跃的病毒复制；干扰素对丙肝病毒及复制活动有抑制作用。负链RNA作为病毒复制活动的直接标志，对评估和预测丙肝治疗的疗效有一定的意义。     

丙型肝炎的发病机制

实验证明 ,HCV感染后 ,即可表现为急性肝炎 ,也可表现为慢性携带状态 ,慢性迁延性肝炎 ,慢性活动性肝炎和肝硬变 ,甚至肝癌。对这种临床表现复杂 ,慢性化倾向高 ,中和抗体水平低或不产生 ,并与原发性肝癌有较为密切关系的疾病 ,发病机制的研究应引起重视。1　直接致病作用丙肝患者血清 HCV- RNA含量和 AL T呈正相关 ,说明病毒的复制常伴随肝损伤 [1 ]。丙肝患者血浆 ,外周血单个核细胞中仅存在正链 HCV- RNA,而肝细胞中除正链HCV- RNA外 ,还存在着复制中间体即负链 RNA,提示肝细胞是 HCV复制的主要场所 ,也是肝细胞损害的重要原…     

六月青含药血清对HepG2.2.15细胞HBV复制的抑制作用

目的:研究六月青(LYQ)含药血清对HepG2.2.15细胞乙型肝炎病毒(HBV)复制的抑制作用。方法:采用血清药理学方法,在HBV的体外细胞培养系统(HepG2.2.15细胞)中观察LYQ抗HBV的作用。结果:LYQ含药血清在HepG2.2.15细胞培养中可有效地抑制HBV DNA的复制,其作用呈显著的浓度-效应和时间-效应关系。结论:LYQ在体外能显著抑制HBV,可能是复方六月雪主要活性成分之一。     

莪术提取物对肝癌细胞系HepG2的抗癌作用及机制研究

目的制备莪术提取物莪术油(zedoary turmericoil)和莪术醇(curcumol)的脂质体,并研究其对肝癌细胞系HepG2的作用及机制,为临床上利用莪术提取物治疗肝癌提供理论依据。方法利用机械分散和超声乳化法制备莪术醇脂质体和莪术油脂质体。通过MTT法观察莪术醇和莪术油对HepG2细胞生长的影响;用Hoechst33258/PI双染色、激光共聚焦法观察细胞凋亡情况;采用RT-PCR法测定HepG2细胞中COX-2 mRNA和VEGF mRNA表达情况。结果莪术醇脂质体和莪术油脂质体的包封率分别为86.2%和74.5%。MTT测定结果显示,莪术醇和莪术油都可抑制HepG2细胞增殖。同时,莪术醇和莪术油都能明显引起HepG2细胞凋亡。与对照组相比,莪术醇81mg·L-1剂量组的VEGF mRNA表达量明显减少,莪术醇27、81mg·L-1剂量组的COX-2 mRNA的表达量明显减少。结论莪术提取物能明显抑制体外培养的HepG2细胞生长,并可诱导其凋亡,其机制可能是通过抑制HepG2细胞COX-2和VEGF基因表达而发挥作用。     

丙型肝炎患者血清HCV-RNA载量与抗-HCV及丙氨酸氨基转移酶联合检测的意义

目的:探讨丙型肝炎患者血清HCV-RNA载量与抗-HCV及丙氨酸氨基转移酶(ALT)浓度的关系。方法:采用实时荧光定量PCR法检测HCV-RNA载量,酶联免疫吸附法(ELISA)检测抗-HCV,全自动生化分析仪连续监测法测定ALT活性浓度。结果:289例丙型肝炎患者的血清标本中,抗-HCV均为阳性,282份标本HCV-RNA均高于检测上限(1 000IU/mL),两种检测方法的符合率为97.6%。ALT异常率随着HCV-RNA载量的增高而升高。HCV-RNA载量与ALT异常率呈正相关(r=0.968,P<0.05)。而ALT浓度变化与HCV-RNA载量并无相关性(r=0.028,P>0.05)。结论:HCV-RNA载量与抗-HCV检测是诊断HCV感染的重要指标,HCV-RNA载量是反映HCV复制的可靠指标,结合丙氨酸氨基转移酶测定,可以帮助临床了解HCV在体内复制的状况及肝脏的损伤情况,为临床诊断HCV感染和评价抗HCV治疗效果提供可靠依据。     

苦参碱的体外抗乙型肝炎病毒作用

目的:观察苦参碱的体外抗HBV作用,初步探讨其作用机理。方法:4×10~4个/mL有HBV基因组的HepG2 2.2.15细胞接种于24孔板,37℃5%CO_2培养24h收取上清液后,更换培养基并加入0.025-0.5mg·mL~(-1)的苦参碱。每3d收取上清,换原浓度药液继续培养,共9天。用酶联免疫法测定上清中HBsAg和HBeAg,酚氯仿抽提被裂解细胞中的HBV DNA,用荧光定量PCR法测定细胞内及细胞外HBV DNA的拷贝数。结果:0.025-0.5mlg·mL~(-1)的苦参碱给药9d,对HepG2 2.2.15细胞分泌HBsAg的半数抑制浓度(IC_(50))为0~(-1)14 mg·mL~(-1),选择指数(SI)为5.66;对HBeAg的IC_(50)和S1分别为0.29mg·mL~(-1)和2.69;对细胞内HBV DNA的IC_(50)和SI分别为0.109mg·mL~(-1)和7.0;对细胞外HBV DNA的IC_(50)和SI则分别为0.120mg·mL~(-1)和6.39。结论:体外实验表明:在0.25-0.5mg·mL~(-1)的剂量范围内,苦参碱对2.2.15细胞中人乙型肝炎病毒的复制有不同程度的抑制作用。     

乙酰吉他霉素临床前药理学研究

乙酰吉他霉素对临床分离的革兰氏阳性球菌有较好的抑菌活力，其对金葡球菌、β－溶血性链球菌、表葡球菌的ＭＩＣ_（５０）分别为１、０．２２和４ｍｇ／Ｌ，对耐红霉素、青霉素的金葡球菌、表葡球菌半数以上较敏感，与吉他霉素相似，但其对革兰氏阴性菌无明显作用。乙酰吉他霉素对小鼠实验性细菌感染有明显保护作用。对金葡球菌、肺炎双球菌感染小鼠口服用药的ＥＤ_（５０）分别为７９．６和２５．１ｍｇ／ｋｇ。其疗效与吉他霉素、麦白霉素、乙酰螺旋霉素相似。乙酰吉他霉素小鼠１次口服的ＬＤ_（５０）＞１５ｇ／ｋｇ，与吉他霉素相比毒性无差异。     

3,4-Methylenedioxymethamphetamine enhances the release of acetylcholine in the prefrontal cortex and dorsal hippocampus of the rat

Rationale The neurochemical effects produced by acute administration of 3,4-methylenedioxymethamphetamine (MDMA) on the monoaminergic systems in the brain are well documented; however, there has been little consideration of the potential effects of MDMA on other neurotransmitter systems. Objective The present study was designed to investigate the acute effect of MDMA on cholinergic neurons by measuring acetylcholine (ACh) release in the medial prefrontal cortex (PFC) and dorsal hippocampus, terminal regions of cholinergic projection neurons originating in the basal forebrain. Methods In vivo microdialysis and high-performance liquid chromatography with electrochemical detection (HPLC-ED) were used to assess the effects of MDMA on the extracellular concentration of ACh in the PFC and dorsal hippocampus of the rat. Results The systemic administration of MDMA (3–20 mg/kg, i.p.) resulted in an increased extracellular concentration of ACh in the PFC and dorsal hippocampus. Reverse dialysis of MDMA (100 μM) into the PFC and hippocampus also increased ACh release in these brain regions. Treatment with parachlorophenylalanine and α-methyl-para-tyrosine, inhibitors of serotonin (5-HT) and dopamine (DA) synthesis, respectively, significantly attenuated the release of ACh stimulated by MDMA in the PFC, but not in the dorsal hippocampus. Conclusions MDMA exerts a stimulatory effect on the release of ACh in the PFC and dorsal hippocampus in vivo, possibly by mechanisms localized within these brain regions. In addition, these results suggest that the MDMA-induced release of ACh in the PFC involves both serotonergic and dopaminergic mechanisms.     

Impact of sampling time deviations on the prediction of the area under the curve using regression limited sampling strategies

The regression limited sampling strategy approach (R‐LSS), which is based on a small number of blood samples drawn at selected time points, has been used as an alternative method for the estimation of the area under the concentration–time curve (AUC). However, deviations from planned sampling times may affect the performance of R‐LSS, influencing related therapeutic decisions and outcomes. The aim of this study was to investigate the impact of different sampling time deviation (STD) scenarios on the estimation of AUC by the R‐LSS using a simulation approach. Three types of scenarios were considered going from the simplest case of fixed deviations, to random deviations and then to a more realistic case where deviations of mixed nature can occur. In addition, the sensitivity of the R‐LSS to STD in each involved sampling point was evaluated. A significant impact of STD on the performance of R‐LSS was demonstrated. The tolerance of R‐LSS to STD was found to depend not only on the number of sampling points but more importantly on the duration of the sampling process. Sensitivity analysis showed that sampling points at which rapid concentration changes occur were relatively more critical for AUC prediction by R‐LSS. As a practical approach, nomograms were proposed, where the expected predictive performance of R‐LSS was provided as a function of STD information. The investigation of STD impact on the predictive performance of R‐LSS is a critical element and should be routinely performed to guide R‐LSS selection and use. Copyright © 2015 John Wiley & Sons, Ltd.     

Activity of the enzymes of the antioxidative system in cadmium-treated Oxya chinensis (Orthoptera Acridoidae)

One purpose in this research was to determine the toxic effects of Cd on antioxidant enzymes of Oxya chinensis (Orthoptera: Acridoidae). Changes in the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and guaiacol peroxidase (GPx) were measured in O. chinensis insects injected with Cd²⁺. Fifth-nymphs of O. chinensis insects were injected with Cd²⁺ at different concentrations (0, 0.55 × 10⁻⁴, 1.10 × 10⁻⁴, 1.65 × 10⁻⁴, 2.20 × 10⁻⁴, and 2.75 × 10⁻⁴ g g⁻¹). An increase in SOD activity in O. chinensis was observed at 1.10 × 10⁻⁴ to 2.75 × 10⁻⁴ g g⁻¹ Cd²⁺. The SOD activity was lower at 2.20 × 10⁻⁴and 2.75 × 10⁻⁴ g g⁻¹ than that at 1.10 × 10⁻⁴ and 1.65 × 10⁻⁴ g g⁻¹. It appears that SOD had a positive protective effect at low Cd²⁺ concentrations, and that this effect disappeared at high Cd²⁺ concentrations. CAT activity was accelerated to varying degrees at 1.10 × 10⁻⁴ to 2.75 × 10⁻⁴ g g⁻¹ for males and at 1.10 × 10⁻⁴, 2.20 × 10⁻⁴, and 2.75 × 10⁻⁴g g⁻¹ for females. CAT showed a strong detoxification effect with all treatments. GPx activity decreased with increasing Cd²⁺ concentration with all treatments for males and at 2.20 × 10⁻⁴ and 2.65 × 10⁻⁴g g⁻¹ for females. We showed that GPx activity had a weak detoxification function with all treatments for males and at high Cd²⁺ for females. Thus, CAT had a strong detoxification effect, whereas SOD had a medium and GPx had a weak detoxification effect. Among the three enzymes, CAT played an important role in the damaging mechanisms of reactive oxygen species in O. chinensis insects. Alterations of the antioxidant enzyme level under environmental stresses are suggested as indicators of biotic and abiotic stress.     

维生素K_3对大白鼠脑内亮氨酸脑啡肽含量及豚鼠回肠收缩的影响

临床实践及动物实验均表明ＶｉｔＫ３有镇痛作用。本实验发现ＶｉｔＫ３（６４ｍｇ·ｋｇ－１．ｉｐ）急性用药可使Ｗｉｓｔａｒ大白鼠脑内亮氨酸脑啡肽含量在尾壳核升高４３％（Ｐ＜０．０１）．在下丘脑升高４５％（Ｐ＜０．０５）。ＶｉｔＫ３（１．９６×１０－５～６．２９×１０－４ｍｏｌ．Ｌ－１）剂量依赖性地部分抑制低频电刺激引起的豚鼠回肠收缩，其效价约为吗啡的１／５０００。上述结果提示ＶｉｔＫ３的中枢镇痛作用可能与某些脑区内脑啡肽增多有关。     

防治痴呆新药的构效关系研究

阿尔茨海默病、帕金森病和血管性痴呆是严重危害老年人生命健康的多发病。目前临床应用的防治痴呆药物的作用机制主要涉及改善胆碱能神经递质、激动多巴胺(DA)受体、阻滞N-甲基-D门冬氨酸(NMDA)受体和改善脑血管功能等。综述了近年来报道的防治痴呆新药的构效关系研究进展。     

Comparison of the inhibition potential of parthenolide and micheliolide on various UDP-glucuronosyltransferase isoforms

Abstract

1. Parthenolide (PTL) and micheliolide (MCL) are sesquiterpene lactones with similar structures, and both of them have been reported to exhibit multiple biochemical and pharmacological activities. This study aims to investigate the inhibition of these two compounds on the activity of UDP-glucuronosyltransferases (UGTs).

2. In vitro incubation mixture for recombinant UGTs-catalyzed glucuronidation metabolism of 4-methylumbelliferone (4-MU) was utilized to investigate the inhibition potential. Inhibition kinetics (including inhibition type and parameters) were determined, and in silico docking was employed to elucidate the inhibition difference between PTL and MCL on UGT1A1.

3. MCL showed no inhibition toward all the UGT isoforms, and PTL showed strong inhibition toward UGT1A1. The half-maximal inhibitory concentration (IC₅₀) of PTL on the activity of UGT1A1 was determined to be 64.4?μM. Inhibition kinetics determination showed that PTL exerted noncompetitive inhibition toward UGT1A1, and the inhibition kinetic constant (K_i) was determined to be 12.1?μM. In silico docking method has been employed to show that hydrogen bonds between PTL and the activity cavity of UGT1A1 contributed to the stronger inhibition of PTL on the activity of UGT1A1 than MCL. In conclusion, PTL can more easily induce drug–drug interaction (DDI) with clinical drugs mainly undergoing UGT1A1-catalyzed glucuronidation.

     

大鼠溃疡性结肠炎模型的实验研究

目的对不同剂量的三硝基苯磺酸（ＴＮＢＳ）引起的大鼠溃疡性结肠炎（ＵＣ）模型进行观察和评价。方法采用一次性直肠注入大鼠ＴＮＢＳ（２５～１５０ｍｇ·ｋｇ－１）的３０％乙醇溶液，引起慢性炎症性肠疾病（ＩＢＤ），３ｗｋ后外死动物对各剂量下动物结肠的重量、髓过氧化物酶（ＭＰＯ）活性及组织形态学变化进行观察和评价。结果ＴＮＢＳ在１００～１５０ｍｇ·ｋｇ－１剂量下引起的ＵＣ肠壁明显增厚，炎症和溃疡至少维持７ｗｋ时间，ＭＰＯ活性值显著性升高，组织学检查发现粘膜及粘膜下层有大量中性粒细胞及淋巴细胞、巨噬细胞、纤维细胞浸润，肉芽组织及隐窝脓肿形成，５０ｍｇ·ｋｇ－１剂量时有一较轻度的损伤。２５ｍｇ·ｋｇ－１时对结肠的重量、ＭＰＯ活性及损伤指数都没有显著性改变（Ｐ＞０．０５）。结论用ＴＮＢＳ引起大鼠实验性ＵＣ，其溃疡和炎症维持一较长时间，这一病理特征为炎症性疾病防治药物的研究提供了条件；本模型的最佳剂量为１００ｍｇ·ｋｇ－１左右     

LPS致热家兔诱导HSF1聚合对体温及脑内AVP含量的影响

目的探讨热休克因子1(HSF1)在脂多糖(LPS)致家兔发热过程中的作用及机制。方法70只家兔随机分为4组:正常对照组(N)、槲皮素组(Q)、脂多糖组(L)和槲皮素+脂多糖组(Q+L)。连续观察体温变化;Westernblot方法检测不同实验条件下下丘脑HSF1和热休克蛋白70(HSP70)的表达;放射免疫法检测下丘脑及腹中隔区(VSA)精氨酸加压素(AVP)含量的变化。结果①各组体温变化最大值(△Tmax)由低至高顺序为:Q组相似文献   

脑脊液转铁蛋白及免疫球蛋白含量变化在小儿中枢神经系统感染时的临床意义

目的　通过对小儿中枢神经系统感染时Tf、Ig在脑脊液中含量的变化及其相关性分析 ,为鉴别诊断及更切实的判定病情和估计预后提供参考依据。方法　应用速率散射比浊法测定脑脊液中Tf、Ig。结果　 (1 )三组急性期脑脊液中Tf、Ig含量均高于恢复期。 (2 )结脑组Tf、IgG增高明显 ,化脑组IgM增高明显。病脑组变化最小。 (3)恢复期脑脊液中Tf、Ig含量与对照组比较 ,结脑中Tf、IgG、IgM高于对照组 ,化脑和病脑组IgM均高于对照组 ,Tf、IgG均无差异 ,各组IgA与对照组相比无显著性差异。 (4)病脑急性期Tf与IgG呈正相关 ,恢复期与IgA呈正相关 ;化脑组急性期Tf与IgM、IgG呈正相关 ,恢复期与IgM呈正相关 ;结脑急性期Tf与IgA、IgM均呈正相关 ,恢复期与IgG呈正相关。结论　 (1 )Tf与IgG是诊断结脑的良好指标 ,IgM是诊断化脑的良好指标 ,当脑脊液中IgM >30mg L时 ,可基本除外病脑 ,当脑脊液中IgM <60mg L时 ,化脑的可能性较小。 (2 )Tf与Ig一样 ,可作为判断中枢神经系统感染时血脑屏障及脑损伤与恢复程度的重要指标