不同级别实验动物用于过敏反应检查的对比研究

目的：评价不同级别实验动物用于过敏反应和类过敏反应检查的适用性和灵敏性.方法：按2010年版《中国药典》过敏反应检查法相关规定进行试验.结果：对硫酸鱼精蛋白、骨瓜提取物和卵清蛋白进行过敏反应检查,普通级豚鼠对卵清蛋白过敏反应阳性,对硫酸鱼精蛋白和骨瓜提取物过敏反应阴性,无类过敏反应;SPF级豚鼠对卵清蛋白过敏反应阳性,对硫酸鱼精蛋白有类过敏反应,对骨瓜提取物过敏反应阴性,无类过敏反应.结论：普通级和SPF级豚鼠都适用于过敏反应检查,SPF级豚鼠对类过敏反应敏感.     

Effects of Cydonia oblonga Miller extracts on blood hemostasis,coagulation and fibrinolysis in mice,and experimental thrombosis in rats

Introduction

Cydonia oblonga Miller (COM) is traditionally used in Uyghur medicine for the prevention of cardiovascular disease. The present study is designed to explore the effects of COM extracts on models and markers of thrombosis and related biomarkers.

Materials and methods

20, 40, 80 mg/kg/day COM aqueous extracts and 5 mg/kg/day aspirin, orally for 14 days were compared to untreated controls in mice on bleeding and clotting times, using the tail cutting and glass slide methods and for death rates in collagen–epinephrine pulmonary thrombosis, thrombolysis in vitro and euglobulin lysis time (ELT). In rats, common carotid artery FeCl₃-induced thrombus and inferior vena cava thrombosis occlusion time, plasma concentrations of thromboxane B₂ (TXB₂) and 6-keto-prostaglandine F₁α (6-keto-PGF1α) were measured.

Results and conclusion

Compared to controls, COM extracts dose-dependently prolonged bleeding by 2.17, 2.78 and 3.63 times, vs. aspirin 2.58, and the clotting time by 1.44, 2.47 and 2.48 times, vs. aspirin 1.91. COM reduced pulmonary embolus mortality by 27, 40 and 53%, vs. 47% for aspirin. COM dose-dependently increased thrombolysis by 45, 55 and 63%, vs. 56% for aspirin, and shortened ELT to 71, 61 and 43%, vs. 43% for aspirin. In rats, venous occlusion time was prolonged. Arterial and venous thrombus weights were dose-dependently reduced in COM groups. TXB₂ decreased and 6-keto-PGF1α increased with COM and aspirin, with an association between 6-keto-PGF1α/TXB2 and arterial or venous thrombus weight for all products, and for occlusion time with COM but not for aspirin.

Conclusion

We confirm the experimental effects of COM on hemostasis and thrombosis. Further exploration of putative clinical effects appear justified.     

Infectious bursal disease virus rescued efficiently with 3′ authentic RNA sequence induces humoral immunity without bursal atrophy

A reverse genetics infectious bursal disease virus (RG-IBDV) that contained authentic 3′ RNA sequence was characterized both in vitro and in vivo. LP1-IBDV, a cell line-adapted IBDV strain variant E (VE) was used as the parent virus for constructing viral cDNA clones. Authentic 3′ RNA sequence was generated by using cis-acting hepatitis delta virus ribozyme (HDR). The absence of HDR in the clones did not affect viral protein expression, but the obtained viral titers were reduced by 200-folds when compared to the clones with HDR sequence. RG-IBDV generated from clones with HDR sequence was similar to LP1-IBDV by plaque size, growth kinetics and electron microscopic morphology. RG-IBDV did not cause bursal atrophy in 3-week-old chickens at 3, 7 and 17 days post infection (DPI) and induced high ELISA and neutralizing antibody titers to IBDV at 7 and 17 DPI. The results indicated that RG-IBDV can be generated efficiently with an authentic 3′ RNA terminus and the obtained RG-IBDV is non-pathogenic and immunogenic with the potential as a vaccine strain that can be further genetically modified to broaden its application.     

北京地区人群对防晒霜的认知及使用情况

目的了解北京地区人群对防晒霜的认知及使用情况。方法采取问卷调查方式,进行面对面交谈,并填写调查问卷。结果80.3%(578/720)的人使用过防晒霜,女性明显高于男性(P<0.001)。开始使用防晒霜的年龄为8～60岁,平均28.4±22.0岁,平均使用时间8.1±21.7年。35.8%的男性、34%的中老年人、44.6%的初等文化程度及28.3%的Ⅳ,Ⅴ型皮肤的人群从未使用过防晒霜。女性及Ⅰ,Ⅱ型皮肤的人使用防晒霜比较规律。63.1%的人群使用防晒霜SPF值在15～30之间。17.3%的人不知道自己使用防晒霜的SPF值,以男性、中老年、初等文化者以及Ⅳ、Ⅴ型皮肤人群为主。有一半以上的人不知道防晒指数的意义。结论北京地区人群中使用防晒霜的比例较高。在对防晒霜的认知和使用,不同人群差异较大。     

实时荧光定量PCR技术在SPF鸡四种垂直传播疾病监测中的应用

目的探讨荧光定量PCR检测技术对SPF鸡四种垂直传播病毒的检测应用。方法采集60份SPF鸡及70份普通鸡群蛋清、泄殖腔试子样品,提取样品核酸,分别进行ARV、REV、CAV、ALV四种病毒实时荧光定量PCR检测,根据标准曲线及溶解曲线分析判读样品病毒拷贝数。结果 SPF鸡ALV 2份阳性,检出率3.3%,其余病毒检测均为阴性;普通鸡样品REV检测2份阳性,检出率2.9%,ALV 10份阳性,检出率14.3%。结论荧光定量PCR检测方法最低可检测到100个拷贝核酸,检测灵敏度较高,有望应用于SPF鸡临床样品的病原检测。     

二氧化钛微囊防晒霜的研制

目的 制备含纳米二氧化钛的聚电解质微囊防晒霜,并考察制剂的结构和防晒性能。 方法 以碳酸钙粒子为核心,通过静电吸附原理将聚电解质4-苯乙烯磺酸钠和纳米二氧化钛交替逐层包裹在核心表面形成微囊,加入辅料,按常规方法制备防晒霜。利用扫描电镜和共聚焦显微镜观察微囊结构;防晒系数分析仪测定制剂的防晒性能。 结果 二氧化钛微囊外观圆整,平均粒径为(5.46±1.05) μm,微囊防晒霜防晒SPF值为30.31±2.56。 结论 含纳米二氧化钛的聚电解质微囊防晒霜防晒的效果优于普通二氧化钛防晒霜。     

CYP2F2-generated metabolites, not styrene oxide, are a key event mediating the mode of action of styrene-induced mouse lung tumors

Styrene induces lung tumors in mice but not in rats. Although metabolism of styrene to 7,8-styrene oxide (SO) by CYP2E1 has been suggested as a mediator of styrene toxicity, lung toxicity is not attenuated in CYP2E1 knockout mice. However, styrene and/or SO metabolism by mouse lung Clara cell-localized CYP2F2 to ring-oxidized cytotoxic metabolite(s) has been postulated as a key metabolic gateway responsible for both lung toxicity and possible tumorigenicity. To test this hypothesis, the lung toxicity of styrene and SO was evaluated in C57BL/6 (WT) and CYP2F2(−/−) knockout mice treated with styrene (400 mg/kg/day, gavage, or 200 or 400 mg/kg/day, ip) or S- or R-SO (200 mg/kg/day, ip) for 5 days. Styrene treated WT mice displayed significant necrosis and exfoliation of Clara cells, and cumulative BrdU-labeling index of S-phase cells was markedly increased in terminal bronchioles of WT mice exposed to styrene or S- or RSO. In contrast, Clara and terminal bronchiole cell toxicity was not observed in CYP2F2(−/−) mice exposed to either styrene or SO. This study clearly demonstrates that the mouse lung toxicity of both styrene and SO is critically dependent on metabolism by CYP2F2. Importantly, the human isoform of CYP2F, CYP2F1, is expressed at much lower levels and likely does not catalyze significant styrene metabolism, supporting the hypothesis that styrene-induced mouse lung tumors may not quantitatively, or possibly qualitatively, predict lung tumor potential in humans.     

SPF32629A and SPF32629B: enantioselective synthesis, determination of absolute configuration, cytotoxicity and antibacterial evaluation

We report herein an efficient enantioselective synthesis of SPF32629A and SPF32629B through one-pot enantioselective reduction and protecting-group-free regioselective O-acylation strategy. The absolute configuration of the enantiomerically pure isomers was established by Mosher ester analysis. The inhibitory potencies of the synthesized compounds were assayed in vitro against a panel of microorganisms and against A549 human lung adenocarcinoma cell line. Compounds 2, 11 and 12 displayed moderate to potent antibacterial activity against all the tested strains and compounds 7, 8, 2, 11 and 12 exhibited significant cytotoxicity in a dose-dependent manner with an IC₅₀ values ranging from 2.92 to 4.14 μg/ml and 8-11 μM.     

通用引物SPF1/GP6++与SPF1/GP6+聚合酶链式反应检测多型别人乳头瘤病毒敏感度的比较

目的 对比通用引物SPF1/GP6++与SPF1/GP6+ PCR两种方法检测人乳头瘤病毒（HPV）的敏感度和感染型别范围。方法 以包含HPV16全长DNA序列的质粒为模板,应用HPV L1基因型别特异性引物进行PCR扩增,获得15种型别HPV模拟靶基因序列并克隆入pEASY-T1载体,将梯度稀释的重组质粒掺入50 ng正常人基因组DNA,模拟各型别HPV感染的待测样本,对比SPF1/GP6+和SPF1/GP6++两组通用引物的检测敏感度。进一步在68例人宫颈癌组织DNA样本中进行对比验证。结果 与SPF1/GP6+PCR比较,SPF1/GP6++ PCR对HPV11,31,34,39,51,52,53,56,58,61和66型别的检测敏感度提高了1到2个数量级,对于HPV16,18,33和35常见感染型别的检测敏感度相似。应用SPF1/GP6++ PCR检测宫颈癌组织HPV感染的总阳性率为98.5%（67/68）,检测到11例双重感染和2例三重感染;而应用SPF1/GP6+ PCR检测的总阳性率为95.6%（65/68）,仅检测到7例双重感染。结论 在SPF1/GP6+ PCR基础上建立了SPF1/GP6++ PCR方法,并证实SPF1/GP6++ PCR具有更高的检测敏感度和更广的HPV型别检测范围。