Detection of turkey rhinotracheitis virus in turkeys using the polymerase chain reaction

Six-week-old turkey poults were infected with the virulent UK/3B/85 strain of TRTV. Tracheal and oesophageal swabs were made every 2 to 3 days from groups of five poults and the RNA extracted. The TRTV RNA was then reverse-transcribed into complementary DNA (cDNA) using an oligonucleotide complementary to the 3' end of the fusion protein (F) mRNA. The cDNA was then used in a polymerase chain reaction (PCR) with an upstream primer to generate a product of approximately 0.5 kbp which was detected by ethidium bromide staining after electrophoresis. In this way, TRTV was detected in both types of swab for 17 to 19 days post-infection, nearly 2 weeks after the peak titres of infectious virus. Swabs which were allowed to dry completely before RNA extraction were as successful as swabs kept wet and extracted almost immediately, useful for when samples are collected in the field. The oligonucleotides amplified the 0.5 kbp product from TRTV strains isolated in six countries over a 13-year period, indicating that they might be usable as 'universal' oligonucleotides for TRTV detection.     

Significance of cytogenetic and fluorescence in situ hybridization analysis in evaluating antichronic myeloid leukemia efficacy of different immune effector cells

We report on the antileukemia effect of interleukin 2 (IL2) on different immune cells from 22 patients with chronic myeloid leukemia (CML). Bone marrow cells from these patients were first cultured in modified long-term bone marrow culture medium for several days, then separately cultured with lymphokine activated killer cells (LAK), cytokine-induced killer cells (CIK), and dendritic cell cocultured CIK (DC-CIK) for another 1-2 days. They were then detected for presence of the Philadelphia chromosome (Ph) by cytogenetic analysis and fluorescence in situ hybridization (FISH). The percentage of Ph-chromosome-positive cells in the bone marrow mononuclear cells after culturing with CIK and DC-CIK was significantly lower than that after culturing with IL2 or LAK. Our results demonstrate that cytogenetics and FISH are useful techniques for the evaluation of the anti-CML effect of immune cells and that CIK or DC-CIK can be appropriate candidates for adoptive immune cell therapy in vivo or for leukemia cell purging ex vivo.     

路氏乳杆菌JCM1081黏附相关蛋白的初步鉴定

目的研究路氏乳杆菌(Lactobacillus reuteri,也称罗伊氏乳杆菌)JCM1081菌体表面蛋白对其黏附HT-29细胞的影响。方法将路氏乳杆菌JCM1081菌体进行胰蛋白酶、蛋白酶K处理;用氯化锂和盐酸胍对乳杆菌表面的蛋白进行抽提,进行SDS-PAGE后与黏蛋白受体进行Western blot,并对杂交阳性蛋白进行质谱分析鉴定。结果路氏乳杆菌JCM1081菌体经胰蛋白酶、蛋白酶K处理后,其对HT-29细胞的黏附力显著下降(P<0．01);用氯化锂去除路氏乳杆菌JCM1081菌体外表面的S层蛋白后,路氏乳杆菌JCM1081对HT-29细胞的黏附力无显著变化;Western blot结果显示相对分子质量(Mr)为29×103和14×103的两种菌体表面蛋白与黏蛋白受体杂交中出现了强阳性;质谱分析结果显示29×103蛋白与路氏乳杆菌ATCC55730的h0793蛋白相似性高达71．1％。结论路氏乳杆菌JCM1081菌体表面的蛋白参与了乳杆菌的黏附,其中29×103和14×103的两种胞壁表面蛋白能够特异地识别黏蛋白受体并与之结合,29×103蛋白属ABC转运蛋白家族。     

Deletion of stem-loop 3 is compensated by second-site mutations within the Gag protein of human immunodeficiency virus type 1

Encapsidation of human immunodeficiency virus type 1 (HIV-1) RNA involves specific interactions between viral Gag proteins and viral RNA elements located at the 5' untranslated region (UTR). These RNA elements are termed packaging (psi) or encapsidation (E) signals and mainly comprise the stem-loop 1 (SL1) and SL3 RNA structures. We have previously shown that deletion of the SL1 sequences is compensated by second-site mutations within Gag. Similar studies are now extended to SL3 and the results demonstrate that deletion of this RNA structure is rescued by two point mutations, i.e., A11V in p2 and I12V in nucleocapsid (NC). These two compensatory mutations are different from those associated with the rescue of SL1 deletion, suggesting that SL1 and SL3 may bind to different residues of Gag during viral RNA packaging. Analysis of virion-derived RNA in native agarose gels shows that deletion of SL3 leads to decreases in both viral RNA packaging and dimerization. These defects are corrected by the compensatory mutations A11V and I12V. Yet, defects in viral RNA dimerization at an early stage that were caused by the SL3 deletion in the context of a viral protease-negative mutation cannot be overcome by these two suppressor mutations. Therefore, the positive effects of A11V and I12V on dimerization of the SL3-deleted RNA must have taken place at the maturation stage.     

The study of tuberculosis outbreak in a high school—Shanghai,China, 2017–2018

Journal of Public Health - To investigate the attack rate of active tuberculosis (TB) cases and detection rate of latent tuberculosis infection (LTBI) cases, and to identify possible factors...     

Discrete choice experiment with duration versus time trade-off: a comparison of test–retest reliability of health utility elicitation approaches in SF-6Dv2 valuation

Quality of Life Research - To evaluate and compare the test–retest reliability of discrete choice experiments with duration (DCETTO) and time trade-off (TTO) in the Chinese SF-6Dv2 valuation...     

中医药对糖尿病足溃疡相关细胞因子调控作用的研究进展

糖尿病足溃疡是糖尿病严重的破坏性并发症,致残率及致死率逐年增高,严重威胁人类身心健康。中医药治疗糖尿病足溃疡,注重辨证论治与整体观念相结合,不仅能改善中医证候,更能在加速创面愈合的同时减少创面的复发,一定程度上延缓了糖尿病足溃疡的进一步恶化,降低其致残率及致死率。现代研究发现,糖尿病足溃疡难以愈合与各种细胞因子如炎症因子、生长因子、趋化因子等分布异常密切相关;随着现代医学对中医药研究的不断深入,中药单体及复方调控细胞因子治疗糖尿病足溃疡已成为研究热点,该文通过对目前国内外研究现状进行归纳,得出以下总结：(1)芝麻酚、栀子苷、当归补血汤、紫朱软膏等调节肿瘤坏死因子-α(TNF-α)、白细胞介素（IL）-1、IL-6、IL-10等炎症因子,抑制创面炎症。(2)白芷、丹酚酸B、四效散、壮药拔毒生肌膏等调节血管内皮生长因子（VEGF）、血小板衍生生长因子（PDGF）、转化生长因子（TGF）、表皮生长因子（EGF）等生长因子,促进创面胶原沉积及血管新生。(3)芍药苷、隐丹参酮、蜂毒、回阳生肌汤等调节CXC趋化因子配体（CXCL）1、CXCL2、CC趋化因子配体（CCL）2、CCL3、基质细胞衍生...     

纳洛酮抢救海洛因中毒618例临床分析

目的：观察纳洛酮抢救海洛因中毒的疗效。方法：对618例海洛因中毒患,用纳洛酮抢救,首次剂量给纳洛酮针剂0．4－0．8mg静注,然后以0．4mg每5min1次,直到患清醒后再给0．8－1．2mg维持静滴。结果：618例中死亡9例,抢救成功率达98％。结论：纳洛酮是抢救海洛因中毒的理想特效药。     

恒河猴tPA基因的克隆、测序与真核表达

本试验采用高浓度乙二醇（EG-8）作为胚胎冷冻保护剂,以高浓度的半乳糖作为解冻稀释液,对MMTV-Wnt-1转基因小鼠的早期囊胚进行玻璃化冷冻保存。结果在EG-2中平衡4-6min和解冻时间为3-5min时获得了最佳冷冻效果。此条件下冷冻胚胎的总回收率为88.61%（109/123）;回收胚胎在体外发育率达88.99%（97/109）;孵化率达81.65%（89/109）;发育胚胎的移植出生率为38.36%（69/181）;冷漠胚胎移植出生后经PCR检测一半小鼠整合阳性。结果表明使用高浓度的乙二醇作为冷冻保护剂和以高浓度的半乳糖作为冻稀释液能有效地保存转基因小鼠胚胎。