柴胡加龙骨牡蛎汤改善围绝经期睡眠质量的机制

目的:探讨柴胡加龙骨牡蛎汤有效部位(CHJLMD70E)改善去卵巢合并电刺激小鼠睡眠质量的机制。方法:采用戊巴比妥钠协同睡眠方法,观察CHJLMD70E延长睡眠的用药时间效应;通过酶联免疫法,观察CHJLMD70E对日间和夜间下丘脑-垂体-卵巢轴激素水平变化的作用。结果:CHJLMD70E(48 mg·kg~(-1)),连续灌胃给药7 d,可以明显延长戊巴比妥钠协同睡眠时间(P<0.01);去卵巢合并电刺激小鼠下丘脑促性腺激素(Gonadotropinreleasing Hormone,GnRH)和血清雌二醇(Estradiol,E_2)水平表现出昼夜紊乱,CHJLMD70E可以纠正这种激素水平的节律紊乱。结论:CHJLMD70E可能是通过调节下丘脑-垂体-卵巢轴激素水平节律平衡而改善围绝经期睡眠质量。     

钩藤源活性化合物297和315对慢性不可预知温和应激模型小鼠的影响

目的考察钩藤源活性化合物297和315对慢性不可预知温和应激(CUMS)小鼠的影响。方法应用雄性C57BL/6J小鼠,采用CUMS造模,20 mg·kg~(-1)的297和315分别伴随灌胃给药28 d;采用糖水消耗实验评价小鼠的抑郁样行为,采用高架十字迷宫实验评价小鼠的焦虑样行为,采用水迷宫实验评价小鼠的认知功能。结果 CUMS小鼠与正常小鼠相比,在高架十字迷宫实验中进入开臂时间及次数显著降低(P<0.01),在糖水消耗实验中糖水偏好指数显著降低(P<0.01),在水迷宫实验中进入目标象限的潜伏期显著延长(P<0.05);而化合物297可显著增加CUMS小鼠进入开臂时间及次数(P<0.05)和CUMS小鼠的糖水偏好指数(P<0.05),同时显著缩短CUMS小鼠进入目标象限潜伏期(P<0.05);化合物315对CUMS小鼠焦虑、抑郁样行为和空间学习记忆功能均无显著改善作用。结论 CUMS可造成小鼠的焦虑和抑郁样行为,同时损伤小鼠的空间学习记忆功能;化合物297可显著改善CUMS小鼠的焦虑和抑郁样行为,同时可改善CUMS小鼠的空间学习记忆功能,提示化合物297可能具有抗应激作用。     

不同品系小鼠对mCPP诱导焦虑样行为的差异研究

目的：评价BALB／c和ICR小鼠对1-（3-氯苯基）哌嗪单盐酸盐（mCPP）诱导焦虑样行为的差异。方法：将两品系小鼠各分为四组：生理盐水对照组、mCPP焦虑组、地西泮对照组和地西泮＋mCPP组。行为学检测前30min腹腔注射地西泮、皮下注射mCPP,采用开场实验、明暗穿箱和高架十字迷宫比较两品系小鼠焦虑样行为的差异。结果：地西泮使焦虑的BALB／c小鼠中央路程比值和中央时间比值增加,对ICR小鼠无作用;两品系小鼠运动总路程有差异。地西泮使焦虑小鼠进入暗箱潜伏期延长、明暗穿箱次数增加、暗箱滞留时间缩短,BALB／c小鼠对地西泮的反应比ICR小鼠更敏感。地西泮可使焦虑小鼠开臂进入次数百分比、开臂停留时间百分比和开臂探臂时间百分比增加,两品系小鼠仅在开臂停留时间和开臂探臂时间上有差异。结论：mCPP诱导焦虑小鼠比正常小鼠对地西泮的敏感性高;BALB／c小鼠焦虑水平较低,探究能力和风险评估能力较高,是抗焦虑药物研究中可供选择的较合适的动物品系。     

升麻柴胡复方对幼龄雌性小鼠性成熟的影响

目的:观察升麻柴胡复方水提物对幼龄雌性小鼠性成熟的影响。方法:选用昆明种18~20日龄雌性小鼠,随机分为生理盐水组、戊酸雌二醇组及升麻柴胡复方水提物高、中、低剂量组。实验期间每天观察小鼠阴道开口情况,以阴道开口情况确定给药时间。处死动物,称取小鼠子宫、卵巢组织湿重,并作血清E2含量测定。结果:升麻柴胡复方水提物能提高幼龄小鼠阴道开口率(p0.05),高、中剂量组能显著提高幼龄小鼠子宫、卵巢组织湿重指数(p0.01),但对血清E2含量无明显影响(p0.05)。结论:升麻柴胡复方水提物能促进幼龄雌性小鼠性成熟,改善生殖机能。     

短期服用盐酸氟西汀对昆明小鼠焦虑样行为的影响

目的:盐酸氟西汀是当前一线抗抑郁药物,但是长期服用有副作用.本研究意在探讨短期服用盐酸氟西汀对小鼠焦虑样行为的影响.方法:将昆明小鼠分为实验组(灌胃Flu,20mg/kg)和对照组(灌胃CMC溶液),连续灌胃7d.后用开放场实验和十字高架迷宫实验对小鼠进行测试.结果:实验组小鼠进入开放场中心次数和中心运动距离均少于对照组.进入十字高架开放臂和开放臂停留时间也少于对照组.结论:短期服用盐酸氟西汀加重昆明小鼠焦虑样行为.     

小鼠海马齿状回过表达18ku转位蛋白对焦虑和抑郁样行为的调节作用

目的研究小鼠海马齿状回18 ku转位蛋白(TSPO)过表达对焦虑、抑郁样行为的影响,为TSPO成为药物潜在新靶标提供实验依据。方法成年雄性ICR小鼠海马齿状回每侧微注射携带TSPO基因的慢病毒(LV)(2×108TU·m L-1),以致TSPO过表达(LV-TSPO)。第12,14和16天进行小鼠高架十字迷宫、爬梯实验、明暗箱实验检测焦虑样行为,第18和21天进行小鼠悬尾和强迫游泳行为绝望模型检测抑郁样行为;行为学实验结束,采用Western蛋白印迹法和ELISA方法分别检测海马注射孔周围3 mm组织内TSPO蛋白水平和四氢孕酮含量。结果小鼠海马齿状回微注射慢病毒后12 d,小鼠跨格数和站立次数无明显改变;与LV组相比,LV-TSPO组显著增加小鼠进入中心区时间(14±4 vs 25±12,P<0.05);显著增加小鼠进入开臂次数和开臂时间的百分比〔(13±8)%vs(26±18)%,P<0.05;(6±6)%vs(27±6)%,P<0.05)〕;显著减少小鼠站立次数(21±7 vs 12±5,P<0.05);显著增加小鼠进入明箱的次数和时间〔18±8 vs(26±7)s,P<0.05;72±36 vs(191±90)s,P<0.05〕,提示海马齿状回TSPO过表达小鼠表现出显著的抗焦虑样行为作用;与LV组相比,LV-TSPO组明显减少小鼠在悬尾实验和强迫游泳实验中的不动时间〔(94±33 vs(36±20)s,P<0.01;137±36 vs(90±37)s,P<0.05〕,提示海马齿状回TSPO过表达小鼠表现出显著的抗抑郁样作用。与LV组相比,小鼠海马齿状回TSPO过表达可导致海马注射孔周围3 mm组织四氢孕酮含量显著增加(P<0.05)。结论小鼠海马齿状回过表达TSPO具有显著的抗焦虑、抗抑郁样的行为学效应,且TSPO介导的四氢孕酮水平升高可能是其重要机制。     

复方丹参片对APP/PS1转基因小鼠焦虑样行为的调节作用

目的探讨复方丹参片对阿尔采末病(AD)APP/PS1转基因小鼠焦虑样行为的作用及机制。方法4月龄野生型小鼠(正常组)和APP/PS1转基因小鼠分别灌胃给予溶剂(模型组)或复方丹参片低剂量(180 mg·kg~(-1))、中剂量(360 mg·kg~(-1))、高剂量(720 mg·kg~(-1))和阳性药物舍曲林(15 mg·kg~(-1))60 d后,采用Morris水迷宫实验、新奇抑制摄食实验和高架十字迷宫实验评价复方丹参片对小鼠记忆障碍和焦虑样行为学表现的影响,采用高效液相色谱法(HPLC)和ELISA分别检测小鼠前额皮质、海马和杏仁核γ-氨基丁酸(GABA)和脑源性神经营养因子(BDNF)的含量。结果复方丹参片中、高剂量明显缩短小鼠找到平台的潜伏期,明显增加小鼠原有平台象限的探索时间(P<0.05,P<0.01)。高架十字迷宫试验结果显示,复方丹参片高剂量显著增加APP/PS1转基因小鼠在开臂的时间百分比和进入开臂的次数(P<0.01);新奇抑制摄食实验结果显示,复方丹参片中、高剂量显著缩短APP/PS1转基因小鼠摄食潜伏期(P<0.05,P<0.01)。HPLC和ELISA结果显示,复方丹参片中、高剂量可显著增加APP/PS1转基因小鼠不同脑区如前额皮质、海马和杏仁核GABA和BDNF含量(P<0.05,P<0.01)。结论复方丹参片改善AD转基因小鼠的学习记忆损伤和焦虑样行为,可能与增加脑组织GABA和BDNF水平有关。     

丁螺环酮对吗啡戒断焦虑大鼠海马、杏仁核突触结构可塑性和CaMKⅡ的影响

目的獉獉:探讨丁螺环酮治疗吗啡依赖戒断焦虑的细胞和分子机制。方法獉獉:66只雄性SD大鼠随机分为生理盐水对照组(对照组)、吗啡戒断焦虑组(模型组)、丁螺环酮治疗组(治疗组)。吗啡剂量递增皮下注射10 d自然戒断,于戒断1-3 d丁螺环酮灌胃治疗行高架十字迷宫实验后,取海马CA3区和杏仁核组织,用电镜体视学方法定量检测其各组突触的数密度(Nv)、面密度(Sv)、突触连接带平均面积(S),Western-blot技术检测其CaMKⅡ的含量和磷酸化水平。结果獉獉:与模型组比较,治疗组大鼠进入开放臂的次数和时间明显增加(P<0.01或P<0.05);海马CA3区和杏仁核突触的Nv和Sv显著降低、S增加(P<0.01或P<0.05),CaMKⅡ蛋白含量和β亚基磷酸化水平显著下调(P<0.01或P<0.05)。结论獉獉:逆转吗啡戒断焦虑大鼠海马、杏仁核突触结构的可塑性和CaMKⅡ分子的变化可能是丁螺环酮缓解吗啡戒断焦虑的重要细胞和分子机制。     

不确定性空瓶饮水刺激法和束缚应激法建立小鼠焦虑模型及比较

目的通过比较不确定性空瓶饮水刺激法和束缚应激法诱发小鼠产生焦虑样情绪,筛选一种重复性好、刺激性小且易于操作的方法建立小鼠焦虑模型,为寻找合适的抗焦虑药物的药效学评价模型提供实验依据。方法 SPF级雄性C57BL/6J小鼠采用不确定性空瓶饮水刺激法建立小鼠焦虑模型,实验分为自由饮水组、定时饮水组、生理应激组和空瓶刺激组,每组8只。自由饮水组小鼠全天自由进食饮水;定时饮水组小鼠每天固定2个时间点、间隔12 h,每次饮水10 min;在相同的时间点,生理应激组小鼠每天确保1次饮水10 min,另一次既不给水也不给空瓶;空瓶刺激组小鼠每天饮水时间随机给予1次或2次空瓶进行刺激,共计饮水12次,空瓶16次;实验持续14 d。采用束缚应激法建立小鼠焦虑模型,实验分为正常对照组和束缚应激组,每组8只。正常对照组小鼠正常饲养,束缚应激组小鼠于每天早上9∶00头朝离心管的底部束缚在管内,第1天束缚2 h,随后每天束缚时间延长2 h,直至每天8 h,连续21 d。小鼠在空瓶刺激7和14 d后或束缚应激7,14和21 d后观察体质量的变化。造模结束后,通过旷场实验检测小鼠自主活动和探索行为,高架十字迷宫实验检测小鼠进入开放臂的次数和在开放臂内停留时间;采用ELISA测定束缚应激21 d后小鼠血浆皮质酮(CORT)水平和海马组织中5-羟色胺(5-HT)含量。结果不确定性空瓶饮水刺激法:与自由饮水组相比,定时饮水组小鼠在开放臂内停留时间显著下降(P<0.05);与定时饮水组相比,空瓶刺激7 d后,小鼠体质量显著下降(P<0.05),其他各项行为学指标无明显变化。束缚应激法:与正常对照组相比,束缚应激14 d后,小鼠体质量显著降低(P<0.05),小鼠的自主活动和探索行为无明显变化,进入开放臂的次数和在开放臂内停留时间均显著下降(P<0.05);束缚应激21 d后,小鼠体质量显著降低(P<0.05),血浆CORT含量和海马组织中5-HT含量显著降低(P<0.01,P<0.05)。结论不确定性空瓶饮水刺激法建立小鼠焦虑模型的稳定性和敏感性欠佳,不可控因素较多,刺激14 d未成功建立小鼠焦虑模型。束缚应激14 d可成功建立小鼠焦虑模型,其机制可能与下丘脑-垂体-肾上腺轴功能紊乱和5-HT代谢异常相关。     

应激与去卵巢对小鼠行为活动的影响及中药复方Ⅰ的防治作用

目的 :观察应激和去卵巢造模对小鼠行为活动的影响 ,并观察中药复方Ⅰ对其的防治作用。方法 :本研究首次采用应激和去卵巢造模观察对小鼠睡眠时间、常压耐缺氧时间、爬绳时间及活动次数的影响 ,观察中药复方Ⅰ对其的防治作用。结果 :单纯应激、单纯去卵巢以及应激与去卵巢对小鼠睡眠、耐常压缺氧、爬绳及活动次数有不同程度的影响 ,但应激与去卵巢比单纯应激、单纯去卵巢对小鼠的影响更为强烈。应激和去卵巢可致小鼠睡眠障碍加剧 ,其睡眠潜伏期延长 ,睡眠时间缩短 ;常压缺氧存活时间缩短 ,爬绳时间缩短 ,小鼠活动次数增加。结论 :应激能使去卵巢小鼠对外界环境改变的敏感性增强 ,与目前临床更年期综合征发病机制相符。由酸枣仁、知母等药组成的中药复方Ⅰ可改善应激与去卵巢小鼠的睡眠 ,缩短小鼠入睡潜伏期 ,延长小鼠睡眠时间 ;能增加小鼠体力 ,使其耐常压缺氧存活时间延长 ,爬绳时间延长 ;还具有一定的镇静作用 ,可使小鼠活动次数减少。     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg x kg(-1)) or i.p. (50 mg x kg(-1)) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) 1 x h(-1) x kg(-1) in the male rat and 10.6 (95% CI: 7.5, 15.0) 1 x h(-1) x kg(-1) in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was approximately 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p < 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p < 0.001) in plasma obtained from the male (8.8 +/- 2.0%) compared with the female rat (11.7 +/- 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Changes in angiopoietin expression in glomeruli involved in glomerulosclerosis in rats with daunorubicin-induced nephrosis

AIM: To study the potential pathological role of endogenous angiopoietins in daunorubicin-induced progressive glomerulosclerosis in rats. METHODS: Seventy male Wistar rats were allocated randomly into a daunorubicin group (DRB; n=40) or a control group (n=30). The rats in the DRB group were injected with DRB (15 mg/kg), in their tails. Subsequently, at intervals of 1, 2, 4, 6, 8, and 12 weeks, 5 male Wistar rats in each group were chosen randomly for 24 h urinary protein quantitative measurements (24 h UPQM), and determination of plasma tumor necrosis factor alpha (TNF-alpha), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) levels. Kidney sections were examined by electron microscopy, Periodic Acid Schiff (PAS) staining, immunohistochemical staining and in situ hybridization histochemistry. RESULTS: As glomerulosclerosis progressed in the DRB group, expression of Ang1 mRNA and protein in glomeruli decreased and expression of TNF-alpha protein, Ang2 mRNA and protein in glomeruli increased. Expression of Ang1 mRNA and protein in glomeruli were negatively correlated with 24 h UPQM, Fn protein expression, and mean area of extracellular matrix (MAECM). In comparison, expression of Ang2 mRNA and protein in glomeruli were positively correlated with 24 h UPQM, Fn protein expression and MAECM; furthermore, there was a positive correlation between plasma Ang2 and 24 h UPQM. Plasma TNF-alpha and expression of TNF-alpha in glomeruli were positively correlated with expression of Ang2 mRNA and protein in glomeruli. There was a negative correlation between Ang1 protein expression and Ang2 protein expression in glomeruli. CONCLUSION: During DRB-induced glomerulosclerosis, podocyte injury led to a shift in the balance of Ang1 and Ang2 in glomeruli. Increased TNF-alpha in plasma and glomeruli may upregulate Ang2 expression in glomeruli. Elevated Ang2 in both plasma and glomeruli may mediate protein permeability through the glomerular filtration barrier. Moreover, local expression of Ang2 may facilitate the progress of glomerulosclerosis by upregulating a component expression of extracellular matrix.     

Trichinellosis in immigrants in Switzerland

We describe a case of trichinellosis diagnosed at the Division of Infectious Diseases, Hospital of Lugano, in January 2009. This case was associated with a cluster of cases and was traced to the consumption of contaminated meat after a wild boar hunt in Bosnia.