Comparison of nucleic acid-based detection of avian influenza H5N1 with virus isolation

Nucleic acid sequence-based amplification with electrochemiluminescent detection (NASBA/ECL) of avian influenza virus was compared with viral culture in embryonated chicken eggs. Virus was isolated from blood or anal swabs of chickens artificially infected with highly pathogenic avian influenza A/Chicken/Hong Kong/1000/97 (H5N1). Viral nucleic acid was detected in blood samples by NASBA/ECL immediately prior to death, whilst nucleic acid extracted from anal swabs was detected from the day following artificial infection until death. Thus, blood and/or anal swabs are a suitable source of material for the detection of avian influenza in dead birds, but anal swabs are more suitable for detection of viral genetic material in live birds. Dilution of a known viral standard was used to determine the limit of sensitivity for both NASBA/ECL and egg culture detection methods. The NASBA/ECL method was equivalent in sensitivity to egg culture. The NASBA/ECL results agreed with egg culture data in 71/94 (75.5%) tissue samples obtained from artificially infected birds.     

Eimeria tenella: ginsenosides-enhanced immune response to the immunization with recombinant 5401 antigen in chickens

Three-day-old specific-pathogen-free chickens were subcutaneously immunized with Eimeria tenella recombinant 5401 antigen (100 microg per chicken) with (0.25, 0.5 or 1.0mg per dose) or without ginsenosides, and boosted with the same dosage 14 days later. The chickens were challenged with 6 x 10(4) homologous sporulated oocysts 14 day after the booster. The specific antibody response and lymphocyte proliferation in response to Con A were measured before and 7, 14, 21, 28, 35, 42 days after the immunization. Oocyst output, mortality, and lesion scores were measured to evaluate the protective effects of the immunization. The vaccine containing 0.5 or 1.0mg ginsenosides per dose induces higher antibody response and lymphocyte proliferation in response to Con A than the vaccine without ginsenosides or containing 0.25mg per dose. The oocyst output indicated that recombinant 5401 antigen with ginsenosides (0.5 and 1.0mg per dose) gave a protection rate of 59.38 and 62.5%, respectively. The lesion score in the group vaccinated with recombinant 5401 antigen with 0.5 or 1.0mg ginsenosides per dose were significantly lower than in group without ginsenosides or containing 0.25mg per dose. Therefore, we conclude that ginsenosides have strong adjuvant effects at a dose of 0.5 or 1.0mg when mixed with E. tenella recombinant 5401 antigen, and has a potential as an adjuvant in chicken vaccine.     

中国脊椎动物红色名录

正为全面评估中国野生脊椎动物濒危状况,环境保护部联合中国科学院于2013年启动了《中国生物多样性红色名录——脊椎动物卷》编制工作。通过这项工作,我们编制了《中国脊椎动物红色名录》。本次评估主要依据以下3个标准:(1)IUCN Red List Categories and Criteria(Version 3.1)(IUCN,2012a);(2)Guidelines for Using the IUCN Red List Categories and Criteria(Version 8.1)(IUCN Standards and Petitions Subcommittee,2010);(3)Guidelines for Application of IUCN Red List Criteria at Regional and National Levels(Version 4.0)(IUCN,2012b)。     

Genetic variation in the GCKR gene is associated with non-alcoholic fatty liver disease in Chinese people

Recent genome-wide association studies reported that GCKR rs780094 polymorphism is associated with elevated fasting serum triglyceride levels and elevated levels of C-reactive protein (CRP). There are a ample of data on the association between circulating triglyceride, CRP concentrations and risk of non-alcoholic fatty liver (NAFLD). To determine whether the GCKR rs780094 polymorphism contributes to the development of non-alcoholic fatty liver, a case?Ccontrol study was performed in 903 Chinese subjects. Among study population, 436 patients with B-mode ultrasound-proven NAFLD (318 with steatosis hepatis I°, 90 with steatosis hepatis II° and 28 with steatosis hepatis III°) and 467 controls were genotyped by using TaqMan allelic discrimination assays. We confirmed the association of GCKR rs780094 with NAFLD in Chinese people (OR = 1.607, 95% CI 1.139?C2.271, P _[dom] = 7.2 × 10^?3). In this study, polymorphism in GCKR rs780094 was not significantly associated with the degree of fatty infiltration of the liver. In addition, the T-allele of GCKR rs780094 was significantly associated with increasing fasting triglyceride (P _[add] = 3.8 × 10^?4) and CRP (P _[add] = 2.9 × 10^?4) concentrations after adjusting for age, gender, and BMI. The association with NAFLD remained significant after adjustment for triglyceride, while adjustment for CRP abolished the association. Genetic variation in GCKR gene rs780094 polymorphism contributes to the risk of NAFLD in Chinese people. The effect of genotype on NAFLD is probably mediated through chronic low-grade systemic inflammation rather than through dislipidemia.     

Increased Humoral Immune Responses of Pigs to Foot‐and‐Mouth Disease Vaccine Supplemented with Ginseng Stem and Leaf Saponins

Vaccination is a conventional approach against foot‐and‐mouth disease (FMD) in pigs. However, failure to elicit an immune response to vaccine has been reported. Our previous investigation showed that ginseng stem and leaf saponins (GSLS) and mineral oil acted synergistically to promote Th1/Th2 immune responses to FMD vaccine in mice. This study was designed to i) find the optimal doses of GSLS in oil‐emulsified FMD vaccines to induce immune responses in mice and pigs and ii) to evaluate the effect of oil‐emulsified FMD vaccine supplemented with GSLS on the immune responses in pigs, by measuring the serum indirect hemagglutination (IHA) titer and IgG and IgG subclass levels. The GSLS‐enhanced immune response to FMD oil‐emulsion vaccine depended on the dose of GSLS added to the vaccine. Addition of GSLS at a dose of 40 μg to 2 ml of FMD oil‐emulsified vaccine significantly enhanced the humoral immune responses in pigs, when compared to the vaccine without GSLS (P<0.05). The increased antibodies included IgG1 and IgG2. Hence, GSLS and oil adjuvant synergistically promoted the immune responses to vaccination against FMD in pigs, and GSLS could be a promising vaccine additive to improve oil‐emulsified veterinary vaccines.     

FATP1 Inhibits 11-cis Retinol Formation via Interaction with the Visual Cycle Retinoid Isomerase RPE65 and Lecithin:Retinol Acyltransferase

The isomerization of all-trans retinol (vitamin A) to 11-cis retinol in the retinal pigment epithelium (RPE) is a key step in the visual process for the regeneration of the visual pigment chromophore, 11-cis retinal. LRAT and RPE65 are recognized as the minimal isomerase catalytic components. However, regulators of this rate-limiting step are not fully identified and could account for the phenotypic variability associated with inherited retinal degeneration (RD) caused by mutations in the RPE65 gene. To identify new RPE65 partners, we screened a porcine RPE mRNA library using a yeast two-hybrid assay with full-length human RPE65. One identified clone (here named FATP1c), containing the cytosolic C-terminal sequence from the fatty acid transport protein 1 (FATP1 or SLC27A1, solute carrier family 27 member 1), was demonstrated to interact dose-dependently with the native RPE65 and with LRAT. Furthermore, these interacting proteins colocalize in the RPE. Cellular reconstitution of human interacting proteins shows that FATP1 markedly inhibits 11-cis retinol production by acting on the production of all-trans retinyl esters and the isomerase activity of RPE65. The identification of this new visual cycle inhibitory component in RPE may contribute to further understanding of retinal pathogenesis.     

掌叶半夏有效提取物对宫颈癌细胞株增殖的抑制作用

目的既往的临床研究证实中药掌叶半夏可有效治疗宫颈癌。在此基础上,课题组观察了中药掌叶半夏有效提取物(Pinellia Extract,PE)对宫颈癌细胞株CaSki和HeLa细胞增殖的作用,先前采用MTT法检测细胞活性以及倒置相差显微镜观察细胞形态的变化,初步显示了PE对CaSki和HeLa细胞的增殖有抑制作用;本文将对PE的增殖抑制作用进行进一步的研究。方法采用扫描电镜和透射电镜观察PE作用后CaSki和HeLa细胞表面及内部超微结构的变化,In-cell Western blot检测不同浓度PE作用15min后CaSki和HeLa细胞磷酸化ERK的表达。结果0.15mg/mlPE作用24h后CaSki和HeLa细胞表面结构及内部结构均发生了显著的变化,提示细胞增殖受到抑制;不同浓度PE作用15min后CaSki和HeLa细胞磷酸化ERK的表达随PE浓度的增加逐渐下降。结论PE对CaSki和HeLa宫颈癌细胞株的增殖有明显抑制作用,表现在细胞表面及内部超微结构的变化;增殖抑制作用之一是阻断ERK的磷酸化。实验结果为进一步研究PE的抗宫颈癌作用机制以及新药的研制开发奠定了一定的理论依据和实验基础。     

通过红色名录评估研究中国哺乳动物受威胁现状及其原因

依据中国哺乳类野生种群与生境现状, 我们利用IUCN Red List Categories and Criteria (Version 3.1), Guidelines for Using the IUCN Red List Categories and Criteria和Guidelines for Application of IUCN Red List Criteria at Regional and National Levels (Version 4.0), 评价了中国所有已知的673种哺乳动物的濒危状况。本次评估了71种《IUCN濒危物种红色名录(2015)》没有评估的哺乳动物, 还评估了60种《IUCN濒危物种红色名录(2015)》误认为中国没有分布的哺乳动物。发现中国有3种哺乳动物“野外灭绝”, 3种“区域灭绝”。受威胁中国哺乳动物共计178种, 约占评估物种总数的26.4%, 高于IUCN濒危物种红色名录的物种平均受威胁率(21.8%)。中国哺乳动物1/4的特有种属于受威胁物种。受威胁比例最高的目是灵长目、食肉目与鲸偶蹄目。多数省区的受威胁哺乳动物物种占本省区哺乳动物总数的20-30%。中国哺乳动物种类多分布在中国第二级地理阶梯。生活在高海拔地区的哺乳动物虽然种类少, 但是受威胁哺乳动物的种类比例高。过度利用、生境丧失和人类干扰名列受威胁哺乳动物致危因子的前3位。自从1989年《中华人民共和国野生动物保护法》实施以来, 一些中国濒危哺乳动物的生存状况得到了改善。然而, 鉴于中国哺乳动物区系的独特性和多样性, 以及中国地形地貌的复杂性, 如何拯救这些濒危物种仍是中国生物多样性保护的一项艰巨任务。