毒鲉科棘毒鱼蜇伤大鼠机体损伤机制及治疗的探讨

目的：探讨毒鲉科棘毒鱼（日本鬼鲉）蜇伤的机制．并应用抗氧化剂对毒鱼蛰伤有无治疗或阻断作用。方法：吞噬鸡红细胞法测定被蜇伤大鼠腹腔巨噬细胞的吞噬功能；MTT法检测腹腔巨噬细胞活性；用免疫组化染色方法（SABC方法）检测腹腔巨噬细胞、胸腺和脾的P物质（Substance P，SP）的表达；应用抗氧化剂VitC探讨其对毒鱼蛰伤的治疗作用。结果：日本鬼鲉蜇伤可使大鼠腹腔巨噬细胞吞噬功能及活性明显降低．VitC治疗可以减轻症状。巨噬细胞、胸腺和脾脏SP表达明显增高。结论：蛰伤局部容易感染和难愈合的原因可能与免疫系统损伤密切相关。肢体肿胀、剧痛及胸闷、休克与SP等物质的分泌有关。抗氧化剂VitC有部分治疗作用。     

甘草提取物对人毛乳头细胞增殖和分泌血管内皮生长因子的影响

目的：观察甘草提取物对体外培养的人毛乳头细胞增殖和分泌血管内皮生长因子（VEGF）的影响。方法：采用不同浓度的甘草提取物作用于体外培养的人毛乳头细胞72h，用四甲基偶氮唑蓝（MTT）法测定细胞增殖活性；ELISA法检测细胞培养上清液中VEGF蛋白含量，反转录（RT）-PCR半定量检测细胞中VEGF mRNA表达。结果：与对照组相比，甘草提取物对毛乳头细胞增殖无明显影响；加药组浓度为2．5、5．0、10．0、15．0μg／mL时，甘草提取物促进细胞分泌VEGF，增加细胞内VEGF mRNA表达，差异有统计学意义。结论：甘草提取物对毛乳头细胞增殖无明显影响，但能显著促进毛乳头细胞分泌VEGF，甘草提取物可能是通过增加毛乳头细胞分泌VEGF来促进毛发生长的。     

银杏叶提取物EGb761对氧化应激下白癜风黑素细胞抗氧化作用的影响

目的:明确银杏叶提取物(EGb761)对氧化应激下白癜风黑素细胞抗氧化作用的影响。方法:人正常黑素细胞系PIG1予以H2O2处理建立氧化应激模型,予以不同浓度(50μg/m L、100μg/m L、200μg/m L、300μg/m L、400μg/m L)EGb761处理,采用MTT法检测PIG1细胞活力,生物化学方法检测脂质过氧化代谢产物丙二醛(MDA)、乳酸脱氢酶(LDH),谷胱甘肽过氧化物酶活性及含量,流式细胞仪检测细胞内活性氧(ROS)水平。结果:与模型组比较,EGb761组PIG1细胞活力、SOD、GSHPx水平增高,ROS、MDA及LDH的表达水平降低。结论:银杏叶提取物EGb761可保护黑素细胞抵抗氧化应激损伤。     

金花舒肤胶囊的主要药效学研究

为研究金花舒肤胶囊主要药效学，将Wister大鼠100只，雌雄各半，随机分成5组，分别给予金花舒肤胶囊高、中、低剂量、湿毒清胶囊、生理盐水灌胃给药，1次／d，连续5d。分别观察其透明质酸酶致大鼠足跖肿胀、大鼠毛细血管通透性、豚鼠磷酸组胺致痒阈、小鼠免疫功能的影响及体外抑菌作用。结果：金花舒肤胶囊可使透明质酸酶所致大鼠足跖肿胀度显著减轻（P〈0．01）；显著抑制组胺所致大鼠毛细血管通透性（P〈0．01）；显著提高磷酸组胺对豚鼠的致痒阈（P〈0．01）；对金黄色葡萄球菌等细菌具有显著的抑制作用；促使大鼠胸腺和脾脏萎缩（P〈0．05）；抑制单核巨噬细胞系统对血清中异物的吞噬功能；金花舒肤胶囊对急、慢性炎症有显著抑制作用，可显著抑制瘙痒，抑制机体的免疫功能。证明该制剂可以用于治疗湿疹。     

四黄虎胶囊体外抗Ⅱ型单纯疱疹病毒作用的实验研究

目的：探讨四黄虎胶囊体外对Ⅱ型单纯疱疹病毒（HSV-2）的作用。方法：采用体外细胞培养技术观察四黄虎胶囊对病毒所致细胞病变（CPE）的抑制作用、对病毒感染的预防、直接杀伤和阻止其复制的作用；阳性对照药物阿昔洛韦（ACV）。结果：四黄虎胶囊在浓度为7．8mg／mL时对HSV-2具有抑制细胞病变作用；7．8mg／mL，3．9mg／mL浓度时对病毒感染有预防作用，同时具有降低HSV的感染力、抑制病毒增殖的作用。ACV的抑制浓度为3．12μg／mL。结论：四黄虎胶囊体外具有较好的抗HSV-2作用。     

姜黄素对人单核细胞株THP-1细胞NF—κB、TNF-α和IL-1βmRNA表达的影响

目的：评价姜黄素对人单核细胞株THP-1中炎症相关因子NF—κB、TNF-α和IL-1βmRNA表达的影响。方法：采用佛波酯（TPA）构建鼠耳肿胀急性炎症反应模型,采用不同浓度的姜黄素处理THP-1细胞,1μg／mL脂多糖（LPS）刺激细胞4h,提取细胞总RNA。用RT—PCR方法检测NF-κB、TNF-α和IL-1βmRNA的表达。结果：局部外用0．8和1．6μmol的姜黄素及腹腔注射50mg／kg和100mg／kg姜黄素,对鼠耳肿胀急性炎症反应均有明显抑制作用。浓度为50μg／mL和25μg/mL。的姜黄素可以明显抑制THP-1细胞中LPS诱导的NF—κB、TNF—α和IL-1βmRNA的表达。结论：姜黄素对炎症因子有抑制作用。     

黄芩甙治疗银屑病的机制研究

目的 探讨黄芩甙治疗银屑病的可能机制。方法 以体外培养的角质形成细胞、成纤维细胞、外周血单个核细胞为对象，不同浓度黄芩甙处理细胞后，用MTT比色分析法反映细胞增殖变化，流式细胞仪测定细胞周期分布。结果 1．5～96μg／mL黄芩甙对成纤维细胞显示一定的抑制作用，并呈时间和剂量依赖关系，而该浓度范围对良性角质形成细胞株HaCaT细胞无明显影响，且外周血单个核细胞活力在此浓度不受影响。流式细胞仪检测细胞周期分布变化，成纤维细胞随药物浓度的升高，G0-G1期细胞比例逐渐增高，G2M、S期细胞比例降低；而HaCaT无明显细胞周期分布变化。结论 一定浓度的黄芩甙可通过阻滞成纤维细胞周期发挥抑制增殖作用，进而影响表皮角质形成细胞生长，是其治疗银屑病的可能机制之一。     

顺铂诱导小鼠黑素瘤Cloundman S91细胞株凋亡的研究

目的：探讨顺铂(CDDP）对小鼠黑素瘤Cloundman S91细胞株的作用。方法：采用四甲基偶氮唑蓝(MTT）还原法检测CDDP对S91细胞株存活率的影响，吖啶橙染色观察凋亡细胞形态，流式细胞仪分析凋亡细胞周期，末端脱氧核苷转移酶（TdT）介导的脱氧尿苷三磷酸(dUTP)末端标记法(TUNEL)原位观察，计算细胞平均凋亡指数。结果：S91细胞经CDDP处理后，存活率降低，并与药物浓度呈正相关；荧光显微镜下可见明显的凋亡细胞；细胞周期G1期前出现凋亡峰。CDDP浓度在10、20、30μmol／L时，细胞平均凋亡指数分别为11．5％、19．8％、33．7％。结论：10～30μmol／L的CDDP诱导S91黑素瘤细胞凋亡。     

五倍子等对体外培养的人皮肤瘢痕组织成纤维细胞生长的影响

目的 研究中药对体外培养的人瘢痕组织皮肤成纤维细胞生长的影响，筛选出抗瘢痕有效的中药及其作用浓度。方法 用四甲基偶氮唑盐比色试验和生长曲线测定法，比较单味中药醇提物对肥厚性瘢痕成纤维细胞（Fb）和人正常皮肤Fb的体外生长活力和生长状态的影响。结果 ①五倍子可明显抑制Fb的体外增殖率，并且对肥厚性瘢痕Fb的作用较正常皮肤Fb强。②五倍子生药醇提取物浓度在0－400μg/mL时，对肥厚性瘢痕Fb增殖率的抑制作用与药物剂量呈明显的正相关，其对肥厚性瘢痕Fb的半数抑制浓度为100μg/mL（生药）；在＜400μg/mL浓度时，与秋水仙碱比较，细胞形态无明显改变。③50μg/mL的五倍子可延迟并缩短正常皮肤Fb和肥厚性瘢痕Fb的对数生长期，延长Fb倍增时间。对肥厚性瘢痕Fb和正常皮肤Fb的生长抑制率与药物的作用时间相依赖。结论 中药五倍子对肥厚性瘢痕Fb体外生长增殖的抑制作用明显地与时间和剂量相依赖，为外用该药治疗瘢痕提供了实验依据。     

重组免疫毒素hIL2-LuffinPl联合维A酸对Hut-78细胞的影响

目的：评价重组免疫毒素hIL2-Ltttlin P1联合维A酸乙酯对皮肤T淋巴瘤细胞（Hut-78）增殖、凋亡及细胞周期的影响。方法：将不同浓度hIL2-LttffinP1分别联合10^-7mol／L维A酸乙酯处理Hut-78细胞,用MTT法检测细胞抑制率,流式细胞仪检测细胞凋亡及细胞周期。结果：用浓度为0．5、1、10、20、30、40μmL的hI心-LuflinP1分别联合10^-7mol／L的芳维A酸乙酯作用Hut-78细胞48h,对Hut-78细胞的抑制率分别为21．33±1．65、28．30±2．42、40．58±1．69、51．57±2．54、63．15±2．41、72．454±1．76;用30μ／mL的hIL2-LuffinP1联合10^-7mol／L芳维A酸乙酯分别作用于Hut-78细胞12、24、36、48h,对Hut-78细胞的抑制率为26．96±1．91、38．05±1．64、51．22±0．57、64．30±2．19、72．48±2．23。经浓度为1、30μg／mL的hIL2-LuffinP1分别联合10^-7mol／L的维A酸乙酯作用Hut-78细胞48h,细胞的凋亡率分别为15．94±1．66和34．89±2．11。经30μg／mL的hIL-2-LuffinP1联合10^-7mol／L的维A酸乙酯作用Hut-78细胞48h,主要表现为G1期细胞比例增加和s期减少。结论：hIL2-Luf．6nP1联合维A酸乙酯能够抑制Hut-78细胞的增殖及凋亡。     

Lipid peroxidative potency of photosensitized thiazide diuretics

We examined the lipid peroxidative potency and photohemolytic activity of thiazide diuretics, especially penflutizide (PFZ), to determine the molecular mechanism of thiazide phototoxicity. Ultraviolet A irradiation of squalene in the presence of PFZ, hydrochlorothiazide, methiclothiazide, benzylhydrochlorothiazide, or trichlormethiazide induced in vitro peroxidation as measured by production of the hydroperoxides and thiobarbituric acid-reactive substances. Among the thiazides, PFZ showed the highest potency to photooxidize lipids. PFZ-photosensitized peroxidation of squalene was repressed by the presence of sodium azide or 2,5-dimethylfuran and was accelerated in a D2O suspension. These findings suggest the participation of singlet oxygen in PFZ photoperoxidation of squalene (type II mechanism). PFZ-photosensitized lysis of red blood cells (RBC) accompanied by formation of hydroperoxides in RBC membrane lipids was also noted. These results suggest that membrane lipids can be one of the target molecules of thiazide phototoxicity.     

Natural killer (NK) cell activity and NK-related cell surface markers in patients with atopic dermatitis

Natural killer (NK) cell activity and NK-related surface markers in peripheral blood mononuclear cells (PBMC) in thirty-three patients with adult type atopic dermatitis (AD) were investigated. NK cell activity, detectable by cytolysis of erythroleukemic cell line K562 cells, was reduced in patient PBMC as compared with healthy controls. Imbalances in the lymphocyte populations reactable with the monoclonal antibodies, anti-HNK-1, and anti-CD16 were found in the patient PBMC. Some positive correlations between these cell populations and the NK cell activities were recognized. In most cases, high levels of serum IgE were inversely correlated with NK cell activity. Depletion of the macrophages from patient PBMC had no effect on NK cell activities. These activities were enhanced by culture with β-interferon for 18 hours, as were the NK cell activities of healthy controls. These results suggest that reduced numbers or imbalances in the lymphocyte population with NK cell activity, are responsible for the pathogenesis and the prognosis of adult type atopic dermatitis.     

Leu7 (HNK-1)-positive cells in peripheral blood and natural killer cell activity in patients with atopic dermatitis]

To clarify the immunologic abnormalities in patients with atopic dermatitis (AD), the percentages of Leu7 (HNK-1)-positive cells and natural killer (NK) cell activity were measured in peripheral blood lymphocytes isolated from 25 AD patients (8 males and 17 females, mean age of 20.8 years old) and 69 healthy non-atopic persons (46 males and 23 females, mean age of 27.4 years old). The percentage of Leu7-positive cells was significantly reduced in AD patients compared with that in controls (AD patients: 11.6 +/- 7.0%, controls: 19.5 +/- 8.3%, p less than 0.01). Although the difference was not statistically significant, reduction of the percentage was slighter in patients with mild AD than in moderate and severe AD patients (mild AD: 14.3 +/- 6.6%, moderate AD: 10.1 +/- 4.4%, severe AD: 10.5 +/- 8.3%). There was also a significant inverse correlation between the percentage of Leu7-positive cells and log IgE levels in patients with AD (r = -0.537, p less than 0.01). No differences in NK cells activity or NK cell activities augmented by interferon-beta and interleukin-2 were observed between in AD patients and in controls. These results suggest that the Leu7-positive cells in AD patients may be closely associated with the regulation of serum IgE production.     

Mosquito salivary gland extracts induce EBV-infected NK cell oncogenesis via CD4 T cells in patients with hypersensitivity to mosquito bites

Severe hypersensitivity to mosquito bites (HMB) is characterized by intense local skin reactions and systemic symptoms such as high fever, lymphadenopathy, and hepatosplenomegaly. Patients with HMB often have natural killer (NK) cell lymphocytosis associated with Epstein-Barr virus (EBV) infection. Here we investigated whether mosquito bites have any influence on the oncogenesis of EBV-infected NK cells. We examined six HMB patients with EBV-infected NK cell lymphocytosis. We first demonstrated that CD4+ T cells, but not NK cells, proliferated well in response to mosquito salivary gland extracts (SGE), especially to SGE of Aedes albopictus. When NK cells were cocultured with autologous CD4+ T cells stimulated by mosquito SGE, the expression of viral oncogene latent membrane protein 1 (LMP1) was remarkably enhanced. Next, we stimulated mononuclear cells of the patients with mosquito SGE, and NK cell counts were monitored for 28 d. The counts changed little from initial levels in the culture with mosquito SGE, whereas they decreased steadily in the culture without the extracts. Furthermore, we detected LMP1 mRNA in the skin lesion induced by mosquito SGE. These results suggest that mosquito bites can induce expression of the viral oncogene LMP1 in NK cells via mosquito antigen-specific CD4+ T cells, which is involved in the oncogenesis of NK cells in vivo.     

Erythrocyte damage in mild and severe psoriasis

BACKGROUND: Psoriasis is a common chronic and recurrent inflammatory skin disorder. Oxygen metabolites and proteases released by activated inflammatory cells may induce oxidative and proteolytic damage to plasma constituents and red blood cells (RBCs). RBCs have a limited biosynthesis capacity and poor repair mechanisms. OBJECTIVES: To study RBCs as a potential cumulative marker of oxidative and proteolytic stress in psoriasis, and as a marker of worsening of the disease. METHODS: The study was performed in 70 patients with mild or severe psoriasis and in 40 control individuals. We evaluated total and differential leucocyte count and, as markers of leucocyte activation, plasma elastase and lactoferrin. Besides the basic RBC study (RBC count, haematocrit, haemoglobin concentration and haematimetric indices) we evaluated antioxidant defences (catalase, superoxide dismutase, glutathione peroxidase and selenium), osmotic fragility and reticulocyte count; in the RBC membrane we evaluated lipid peroxidation and susceptibility to lipid peroxidation, membrane fluidity, levels of cholesterol and phospholipids, membrane-bound haemoglobin, band 3 profile and levels of vitamin E; serum levels of bilirubin, total plasma antioxidant capacity, lipid profile and lipid peroxidation were also evaluated. RESULTS: Psoriasis patients showed a rise in leucocytes, mainly neutrophils, which was associated with a rise in elastase and lactoferrin. Patients had a reduced RBC count, antioxidant defences and membrane fluidity, elevated membrane lipid peroxidation, membrane-bound haemoglobin, osmotic fragility and reticulocyte count, and a different band 3 profile. Most of these modifications were enhanced in severe psoriasis. CONCLUSIONS: In summary, our data show that the RBCs are at a lower number in psoriasis patients, and present several changes denoting an enhanced damage and/or ageing process, which seem to be strongly connected with neutrophil activation, oxidative stress and worsening of psoriasis.     

RPR持续阳性梅毒患者的传染性与细胞免疫的相关性研究

目的:检测RPR持续阳性经治梅毒患者外周血淋巴细胞亚群,探讨其传染性与细胞免疫的相关性。方法:应用流式细胞仪检测RPR持续阳性梅毒患者中38例IgM阳性和32例IgM阴性的患者外周血T细胞亚群及NK细胞,并以30例健康人作为对照组。结果:IgM阳性组外周血CD3、CD4和CD8细胞均高于对照组(P<0.05),而NK细胞低于对照组(P<0.001);IgM阴性组外周血CD3、CD4细胞低于对照组(P<0.05),而CD8细胞及NK细胞均高于对照组(P<0.05);两组之间外周血淋巴细胞亚群的检测结果相比,IgM阳性组CD3、CD4细胞高于IgM阴性组(P<0.001),NK细胞低于IgM阴性组(P<0.001),而CD8细胞相比差异无显著性(P>0.05)。结论:RPR持续阳性梅毒患者存在明显的细胞免疫不平衡和免疫抑制;NK细胞显著降低可能是RPR持续阳性且具传染性的主要原因。     

The basophilic histamine liberation test as a diagnostic method in Hymenoptera venom allergy

In 181 patients with systemic reactions to hymenoptera stings as a result of immediate-type allergy to bee or wasp venom confirmed by history, skin testing and demonstration of hymenoptera venom-specific IgE antibodies on radioallergosorbent testing (RAST), in vitro histamine release from peripheral basophilic granulocytes following stimulation with bee or wasp venom was evaluated. Suspensions of washed peripheral leukocytes at a density of 2 x 10(6)/ml were incubated with four different concentrations of hymenoptera venoms. Histamine was measured by spectrofluorometry, and histamine release was calculated as a percentage of the total histamine content. Tests performed with peripheral leukocytes obtained from 18 non-allergic individuals served as controls. In both bee venom allergy and wasp venom allergy the corresponding allergen induced concentration-dependent histamine release. Upper normal limits of histamine release were defined for the venom concentrations used. When these were used as reference values the basophil histamine release test exhibited a specificity of 94% and a sensitivity of 82% in the diagnosis of bee venom allergy, and a specificity of 83% and a sensitivity of 68% in the diagnosis of wasp venom allergy. Since there was no relevant significant correlation between the results of the basophil histamine release test on one hand and the severity of sting reactions and the prick test and RAST results on the other, it seems that the histamine release test determined additional parameters of sensitization. Thus, the method is a valuable adjunct to the in vitro methods available for the diagnosis of hymenoptera venom allergy.     

银屑病患者红细胞CD35和红细胞趋化因子受体的测定

目的研究银屑病患者红细胞CD35及红细胞趋化因子受体(ECKR),探讨红细胞天然免疫功能状况及其在银屑病发病机制中的作用。方法测定新鲜血红细胞对肿瘤细胞的快速天然免疫反应;用流式细胞仪定量测定红细胞CD35;用ELISA法检测白介素8含量,反映红细胞上ECKR的结合活性。结果银屑病患者肿瘤红细胞花环率和红细胞CD35定量均明显高于对照组(P<0.05),ECKR结合活性比正常人明显降低(P<0.05),红细胞CD35定量及ECKR结合活性与PASI评分呈显著正相关(r =0.93,P<0.001,r =0.49,P<0.005)。结论银屑病患者红细胞天然免疫功能存在亢进和紊乱,红细胞CD35分子数量及其活性的增加是银屑病天然免疫反应能力亢进的基础;红细胞CD35分子定量及ECKR结合活性的测定可作为判断银屑病病情的客观指标;红细胞参与了细胞因子的调控并在银屑病的发病过程中可能起一定作用。     

未治和血清固定梅毒患者外周血淋巴细胞亚群的比较研究

目的检测未治和血清固定梅毒患者外周血T淋巴细胞亚群及NK淋巴细胞,比较两组患者之间细胞免疫的差异.方法应用流式细胞仪检测40例未治梅毒患者和32例血清固定梅毒患者外周血T淋巴细胞亚群及NK淋巴细胞,并与30例健康人群的检测结果相对照.结果未治梅毒患者CD3、CD4及NK淋巴细胞与对照组的检测结果相比差异无显著性(P＞0.05),而CD8淋巴细胞高于对照组(P＜0.001);血清固定梅毒患者组外周血CD3、CD4及CD8淋巴细胞均低于对照组的检测结果(P＜0.05),而NK淋巴细胞高于对照组(P＜0.001);两组患者外周血CD3、CD4、CD8及NK淋巴细胞相比,差异有显著性(P＜0.05).结论未治和血清固定梅毒患者存在细胞免疫不平衡和免疫抑制.     

早期梅毒血清固定患者淋巴细胞的检测及意义

目的探讨细胞免疫与早期梅毒血清固定的关系。方法流式细胞仪检测30例血清阴转和20例血清固定早期梅毒患者的外周血T淋巴细胞亚群及NK淋巴细胞,并与30例健康人对照。结果血清固定组早期梅毒患者CD3,CD4淋巴细胞低于血清转阴组和正常对照组,而CD8淋巴细胞高于这两组,NK淋巴细胞差异无显著性意义;血清阴转组梅毒患者与正常对照组各淋巴细胞亚群和NK细胞差异无显著性意义。结论早期梅毒血清固定患者机体内细胞免疫受到明显抑制及产生免疫不平衡,可能造成了梅毒螺旋体逃脱机体免疫监视而残存,成为引起血清固定的原因之一。