DCC/HBV治疗性疫苗佐剂有效剂量的初步研究

目的 考察新型治疗性乙肝疫苗佐剂的有效剂量.方法 DC-Chol/HBsAg免疫Balb/c小鼠,ELISA法测血清特异性抗体IgG1和IgG2a水平,ELISPOT法检测免疫后小鼠脾细胞经HBsAg刺激后产生IFN-γ的细胞数.结果 DC-Chol最低有效剂量在0.063～0.125 mg/只小鼠.结论 DCC/HBV治疗性疫苗能同时诱导Balb/c小鼠体液和细胞免疫反应,其佐剂最低有效剂量低于文献报道.     

DCC／HBV治疗性疫苗佐剂有效剂量的初步研究

目的考察新型治疗性乙肝疫苗佐剂的有效剂量。方法DC-Chol/HBsAg免疫Balb/c小鼠,ELISA法测血清特异性抗体IgG1和IgG2 a水平,ELISPOT法检测免疫后小鼠脾细胞经HBsAg刺激后产生IFN-γ的细胞数。结果DC-Chol最低有效剂量在0.063～0.125 mg/只小鼠。结论DCC/HBV治疗性疫苗能同时诱导Balb/c小鼠体液和细胞免疫反应,其佐剂最低有效剂量低于文献报道。     

酵母多糖对免疫功能的影响

目的从酵母菌中提取其多糖成分,探讨酵母多糖对小鼠免疫功能的影响,为酵母多糖的临床应用提供实验依据。方法取小鼠脾脏淋巴细胞,置于96孔细胞培养板中,加入酵母多糖为酵母多糖组和细胞培养液为空白对照组,测定小鼠脾细胞的增殖活性;取细胞上清液,测定细胞因子白细胞介素-2(IL-2)、干扰素-γ(IFN-γ);用流式细胞术(FCM)检测细胞表型,观察酵母多糖对细胞免疫和体液免疫作用。结果实验组细胞增殖活性显著高于对照组(P<0.01);实验组IL-2,IFN-γ的浓度与酵母多糖在一定浓度范围内成剂量依赖关系;酵母多糖即能刺激B淋巴细胞,也能刺激T细增殖。结论酵母多糖可提高小鼠免疫功能,它不仅能提高体液免疫功能,而且能提高细胞免疫功能。     

CpG ODN纳米粒增强免疫抑制小鼠对重组乙肝疫苗的免疫应答

目的：探讨CpG ODN壳聚糖纳米粒联合重组乙肝疫苗对免疫抑制小鼠的免疫增强作用。方法：选月环磷酰胺建立免疫抑制模型小鼠。将乙肝疫苗单独或和CpG ODN或CpG ODN纳米粒经后腿胫骨前肌注射到小鼠体内，ELISA方法检测抗HBsAg IgG抗体、IL-12水平；流式细胞仪检测外周血CD4^＋、CD8^＋T淋巴细胞亚群。结果：CpG ODN纳米粒联合疫苗组的CD4^＋细胞百分数，IL-12水平及抗HBsAg IgG抗体的含量显著高于CpG ODN联合疫苗组（P〈0．05）。结论：包裹在壳聚糖纳米粒中的CpG ODN较相同剂量的CpG ODN更能增强免疫抑制小鼠对乙肝疫苗的免疫应答。     

黄芪多糖对免疫功能影响的体外实验研究

目的探讨黄芪多糖对小鼠免疫功能的影响,为黄芪多糖的临床应用提供理论依据。方法取小鼠脾细胞,置于96孔细胞培养板中,设黄芪多糖组和空白对照组,WST-1法测细胞的增殖活性;ELISA测细胞因子IL-2、IFN-γ;流式细胞术（FCM）检测细胞表型。结果黄芪多糖组细胞增殖活性显著高于空白对照组（P〈0.01）;黄芪多糖组IL-2、IFN-γ的浓度随黄芪多糖浓度的增加而增多,在一定浓度范围内呈剂量依赖关系;黄芪多糖既能刺激B淋巴细胞增殖,也能刺激T淋巴细增殖。结论黄芪多糖可增强免疫功能,它不仅能提高体液免疫功能,而且能提高细胞免疫功能。     

牛蒡寡糖对小鼠淋巴细胞增殖和IL-2、IFN—γ产生的影响

目的：探讨牛蒡寡糖对小鼠淋巴细胞增殖反应和IL-2、IFN-γ产生的影响。方法：取小鼠40只随机分为4组（对照组与实验组）,将牛蒡寡糖用3个不同剂量组250、500、1000mg／（kg·d）给予小鼠灌胃,对照组为0．9％NaCl溶液,连续灌胃14d后,检测小鼠淋巴细胞增殖和IL-2、IFN-γ的产生。结果：牛蒡寡糖能提高小鼠的脾淋巴细胞增殖及IL-2、IFN-γ的分泌。结论：牛蒡寡糖对正常小鼠的免疫功能具有促进作用。     

杨梅素对淋巴细胞活化及增殖的影响

目的研究杨梅素(myricetin)对小鼠T细胞活化及增殖的影响,探讨其作为免疫调节药物开发的可能性并提供相关实验依据。方法无菌分离小鼠淋巴结细胞,加入不同浓度的杨梅素预先孵育1h,用多克隆刺激剂刀豆蛋白(ConA)刺激T细胞活化,利用荧光标记的单克隆抗体双染技术,流式细胞仪检测杨梅素对小鼠CD3+T细胞CD69表达的影响;采用3H-TdR参入法检测杨梅素对淋巴细胞增殖的影响,采用半定量RT-PCR技术从mRNA水平检测杨梅素对IL-2mRNA表达的影响。采用ELISPOT检测杨梅素对IFN-γ分泌的影响。结果杨梅素能抑制T细胞早期活化标志CD69的表达,并能抑制淋巴细胞增殖反应,同时对淋巴细胞活化后IL-2 mRNA的表达及IFN-γ的分泌也有抑制作用。结论杨梅素对淋巴细胞的活化增殖反应具有抑制作用。     

姬松茸多糖ABP-AW1对免疫功能低下小鼠的免疫增强作用

目的探讨低分子质量姬松茸多糖ABP-AW1对免疫功能低下小鼠的免疫增强作用。方法 ICR小鼠ip给予环磷酰胺80 mg·kg~(-1),连续3 d,制备免疫功能低下小鼠模型;随后分别ig给予ABP-AW1 125,250和500 mg·kg~(-1),每天1次,连续7 d。治疗结束后处死小鼠,检测小鼠胸腺指数和脾指数,碳廓清实验检测小鼠巨噬细胞吞噬功能,MTT法检测小鼠脾淋巴细胞增殖反应,ELISA法检测小鼠脾淋巴细胞分泌白细胞介素2(IL-2)和干扰素γ(IFN-γ)水平,流式细胞术检测小鼠脾T淋巴细胞亚群CD4~+/CD8~+比值。结果与正常对照组比较,免疫低下模型小鼠胸腺指数和脾指数以及巨噬细胞吞噬指数均降低(P<0.05);ABPAW1 250和500 mg·kg~(-1)可显著提高免疫低下模型小鼠胸腺指数和脾指数(P<0.05),升高巨噬细胞吞噬指数(P<0.05)。与正常对照组比较,模型小鼠伴刀豆凝集素A(Con A)和脂多糖(LPS)分别诱导的T淋巴细胞增殖反应和B淋巴细胞增殖反应降低,脾细胞分泌IL-2和IFN-γ水平降低,脾细胞CD4~+/CD8~+比值下降(P<0.05);与模型组比较,ABP-AW1 250和500 mg·kg~(-1)可促进免疫低下模型小鼠Con A和LPS分别诱导的T淋巴细胞增殖反应和B淋巴细胞增殖反应(P<0.05),显著增加脾细胞IL-2和IFN-γ水平(P<0.05),明显逆转免疫低下小鼠脾细胞CD4~+/CD8~+比值的下降(P<0.05)。结论 ABP-AW1对免疫功能低下小鼠具有一定的免疫增强作用。     

细胞因子对小鼠肺癌细胞MHC Ⅰ类分子和共刺激分子的调节作用

目的观察T细胞活化因子对小鼠肺癌细胞表达MHCⅠ类分子和共刺激分子的调节作用,探讨细胞因子增强特异性抗肿瘤应答的免疫效果。方法用rhIL-2和IFN-γ分别活化对数生长期的小鼠lewis肺癌3LL细胞48h,FACS检测细胞因子活化前后3LL细胞表达MHCⅠ类分子(H-2)和共刺激分子(CD80)的荧光强度。结果3LL细胞低表达H-2和CD80,经rhIL-2和IFN-γ活化48h后,3LL表达H-2和CD80的荧光强度明显增强,rhIL-2的活化的两种分子荧光强度高于IFN-γ。结论rhIL-2和IFN-γ能上调小鼠lewis肺癌3LL细胞表面MHCⅠ类分子和共刺激分子的表达,增强体内特异性抗肿瘤免疫应答。     

阿糖胞苷刺激Jurkat细胞B7和细胞因子mRNA表达

目的探讨B7共刺激分子在T细胞活化和分化中的作用。方法阿糖胞苷(Ara-C)刺激Jurkat细胞,流式细胞术检测刺激前、后B7-1、B7-2的表达,RT-PCR检测刺激前、后B7-1、B7-2基因的表达,检测T细胞表面因子IL-3和IFN-γ的表达以及这两种细胞因子在Ara-C刺激Jurkat细胞后12,24,48,72,96,120,144 h时间动态变化。结果经Ara-C刺激Jurkat细胞后,均能使B7-1、B7-2表达上调,并且可诱导T细胞分泌细胞因子IL-3和IFN-γ,而未经Ara-C刺激的Jurkat细胞,无B7-1、B7-2的表达,均未诱导细胞因子的分泌。IL-3在T细胞活化12 h即可检测到,在48 h表达量最高,IFN-γ在T细胞活化24 h即可检测到,在72 h表达量最高。结论Ara-C可有效地刺激Jurkat细胞B7的表达,有效地增强肿瘤细胞的免疫原性,激活T细胞。B7在T细胞活化中起着重要作用。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.