Characterization and phylogenetic analysis of a neurovirulent Zika virus isolated from Cambodia in 2019

The geographic range of Zika virus (ZIKV) has expanded from Asia to the Americas, leading to the 2015–2016 pandemic with enhanced neurovirulence. At present, ZIKV is continuously circulating in many Southeast Asian countries. Unfortunately, the persistent evolution of ZIKV in Southeast Asia and its influence on the biological characteristics of the virus remain incompletely understood. In this study, the in vitro and in vivo properties of a new ZIKV isolate obtained from Cambodia in 2019 (CAM/2019) were characterized and compared with those of the Cambodian strain (CAM/2010). Compared with CAM/2010, the CAM/2019 virus showed similar plaque morphology and growth curves in cell cultures and induced comparable viremia and organ viral loads profiles in both BALB/c and A129 (IFNAR1^−/−) mice upon intraperitoneal (i.p.) inoculation. Remarkably, the CAM/2019 virus exhibited enhanced neurovirulence in neonatal mice compared with CAM/2010, with a 74-fold reduction in the 50% lethal dose (LD₅₀). Consistently, CAM/2019 produced higher viral loads in the brains of BALB/c neonatal mice than CAM/2010 did. Sequence alignment showed that the CAM/2019 virus has acquired 12 amino acid substitutions, several of which were found to be associated with neurovirulence. In particular, the CAM/2019 virus shared an A1204T substitution in NS2A with the Thai isolate SI-BKK02 that was isolated from a microcephaly case. Taken together, our results indicate that a ZIKV strain isolated with specific mutations has emerged in Cambodia, highlighting the need for extensive molecular and disease surveillance in Cambodia and other Asian countries.     

古尼拟青霉对小鼠缺血性脑损伤保护作用研究

探讨古尼拟青霉对小鼠急性脑损伤的保护作用。发现古尼拟青霉能显著延长小鼠断头后张口呼吸持续的时间,使小鼠双侧颈总动脉（含迷走神经）不完全结扎后的存活时间明显延长,提示该药可能对缺血性脑损伤有一定的保护作用。     

立体定向微电极导向射频毁损术治疗帕金森病108例

目的 介绍立体一定向微电极导向射叔毁损术治疗帕金森病及术后可能出现的并发症。方法 采用ＣＴ定位立体定向微电极细胞电生理记录技术进行术中靶点监测,对１０８例帕金森病人的苍白球和（或）丘脑腹外侧核进行射频毁损术治疗。结果 手术效果优良,原有症状术后改善率９８．１％,无永久并发症。结论 立体定向微电极导向射频毁损术是目前治疗帕金森现的一种安全有效方法。     

血清HA、PCⅢ对慢性肝炎、肝硬化的诊断价值

目的 探讨血清ＨＡ、ＰＣⅢ对慢性肝炎,肝炎肝硬化的诊断价值,ＨＡ与ＰＣⅢ意相关关系以及它们的变化对预后的影响。方法 采用ＲＩＡ法同时检测慢性肝炎、肝炎肝硬化患者血清ＨＡ、ＰＣⅢ含量,比较ＨＡ,ＰＣⅢ间关系,肝炎肝硬化组中存活病例与死亡病例的ＨＡ、ＰＣⅢ变化情况。     

氟西汀治疗抑郁性障碍三期临床验证

目的：验证国产氟西洒汀长期、大范围治疗抑郁性障碍的疗效和副反应。方法：采用国产氟西汀开放治疗抑郁性障碍30例,以HAMD和TESS评定症状、疗效和副反应。结果：氟西汀对各类型抑郁性障碍的疗效肯定;显效率达80％,副反应少且轻。结论：氟西汀是一种安全、有效、广谱的抗抑郁剂。     

Establishment of the culture technique of pulmonary vascular pericytes and its identification in rats

Summary In order to study the cellular origin of muscularization in non-muscular arterioles of the lung, the pulmonary vascular pericytes-culture was established. The terminal lung tissue of the rat was taken out and minced. Then 0.5 % of type IV collagenase solution was added for digestion and the microvascular segments were obtained by screening. The targeted cells were cultured by “selective conditioned media”. Under phase-contrast microscope, the cultured cells were large in size with ragged margin and numerous pseudopodia, which imparted tubule-like structure. There was no contact inhibition in growing cells, so multiple layers developed. When they were confluent, there were morphologically no “hillock and dale” growth pattern as in smooth muscle cells or “weave-like” pattern as in fibroblasts. The ultrastructure of cultured cells showed numerous digital processes, moderate amount of rough and smooth endoplasmic reticulum, rich Golgi’s apparatus, microfilaments, few lysosomes without myofilaments and dense bodies. Immunohistochemical staining revealed that the cultured pericytes had same kind of cellular skeletal protein, α-SM-actin, like smooth muscle cells. The cultured cells also exhibited positive reaction to CD₃₄ antigen and S-100 antigen, which were negative in smooth muscle cells and fibroblasts. The cell growth pattern, ultrastructure and immunological phenotype suggested that the cultured cells had characteristics of vascular pericytes. Pericytes are one of the components of microvascular cells, and the establishment ofin vitro culture technique of pericytes is of significance for further exploration of the muscularization of non-muscular arterioles in lung and the mechanism of structural remodeling of pulmonary vessels. This project was supported by the grant of National Nature Sciences Foundation of China (No. 39570289).     

Effect of N-acetylcysteine on mitochondrial function following traumatic brain injury in rats

Efficacy of N-acetylcysteine (NAC) in traumatic brain injury (TBI)-induced mitochondrial dysfunction was evaluated following controlled cortical impact injury in rats. Respiratory function and calcium transport of rat forebrain mitochondria from injured and uninjured hemispheres were examined. NAC significantly restored mitochondrial electron transfer, energy coupling capacity, calcium uptake activity and reduced calcium content absorbed to brain mitochondrial membranes when examined 12 h post-TBI if NAC was administered i.p. 5 min before injury or 30 min or 1 h postinjury. Glutathione (reduced form, GSH) levels in brain tissues were decreased at all time points examined over a 14-day observation period, while mitochondrial GSH levels significantly decreased only at 3 days and 14 days following TBI. NAC treatment given within 1 h greatly restored brain GSH levels from 1 h to 14 days and mitochondrial GSH levels from 12 h to 14 days post-TBI. NAC did not show protective effects when given 2 h postinjury. Our data indicate that NAC administered postinjury at an early stage can effectively restore TBI-induced mitochondrial dysfunction and the protective effect of NAC may be related to its restoration of GSH levels in the brain.     

香附水提剂对离体兔肠管活动的影响

目的：探讨香附水提剂对离体兔肠平滑肌的作用。方法：采用Magnus实验装置,使用二道仪描记离体兔肠平滑肌收缩曲线5min作为对照,然后观察药物对其作用25min后曲线变化,并与对照组比较。结果：给香附水提剂后5min离体平滑肌收缩频率及幅度降低较阿托品对照组明显;5-15min二者接近;25min后降低幅度为阿托品组1.3倍,二者比较差异显著（P＜0.01）。结论：香阳水提剂能明显抑制离体兔肠平滑肌的收缩幅度与频率,同时也能拮抗乙酰胆碱和氯化钡所致离体肠管平滑肌的兴奋作用。     

内眼手术前,后结膜囊细菌学研究

目的:探索眼科内眼手术前后结膜囊细菌学变化特点及临床眼科用药效果。方法:133眼需要行内眼手术的连续病例包括白内障手术68眼、青光眼手术39眼、视网膜脱离手术20眼、玻璃体切割术6眼。用133眼内眼手术病人手术前后的结膜刮片及68眼白内障手术术毕的前房冲洗液作细菌培养,并作药敏试验。结果:133眼内眼手术患者入院时结膜刮片作细菌培养有61眼(45.85％)培养出细菌,其中金黄色葡萄球菌23眼、表皮葡萄球菌25眼、G~ 杆菌13眼;手术消毒前结膜囊刮片有8眼(6％)仍培养出细菌,其中金黄色葡萄球菌3眼,表皮葡萄球菌5眼;68眼白内障手术患者前房冲洗液中仅1眼(1.5％)培养出细菌,为表皮葡萄球菌;术毕时结膜刮片4眼(3％)培养出细菌,所培养出来的细菌均对妥布霉素、庆大霉素、先锋Ⅵ、万古霉素和利福平敏感,而对四环素、氯霉素、红霉素、氧氟沙星耐药。结论:肉眼手术前应用有效抗菌素至为重要,可预防眼内炎的发生,但不可轻视手术无菌操作。眼科学报1999;15:267-269。     

Molecular characterization of a new excitatory insect neurotoxin with an analgesic effect on mice from the scorpion Buthus martensi Karsch.

Besides the neurotoxins active on mammals, a new excitatory insect selective toxin with a mice analgesic activity was found and purified from the venom of the scorpion Buthus martensi Karsch (BmK) (Ji, Y.H., Mansuelle, P., Terakawa, S., Kopeyan, C., Yanaihara, N., Hsu, K., Rochat, H., 1996. Toxicon 34, 987; Luo, M.J., Xiong, Y.M., Wang, M., Wang, D.C., Chi, C.W., 1997. Toxicon 35, 723.). This peptide (designated as BmK IT-AP) is composed of 72 amino acid residues. Its primary structure was determined by automated Edman degradation of the N-terminal part of the reduced and S-carboxamidemethylated protein and its lysylendopeptidase degraded fragments. Based on the determined sequence, the gene specific primers were designed and synthesized for 3' and 5' RACE (rapid amplification of cDNA ends). Their partial cDNA fragments obtained by 3' and 5' RACEwere cloned and sequenced and the full length cDNA sequence of BmK IT-AP was then completed by overlapping their two partial cDNA sequences. It encodes a precursor of 90 amino acid residues: a signal peptide of 18 residues and a mature peptide of 72 residues which are consistent with the determined protein sequence of BmK IT-AP. The genomic DNA of the peptide was also amplified by PCR from the scorpion genomic DNA and sequenced, which is a first report on the genomic structure of a scorpion toxin specific for insects. Its sequence revealed an intron of 590 bp inserted in the end part of the signal peptide. The peptide caused a fast excitatory contraction paralysis on house fly larvae. Furthermore, the peptide also showed an obvious analgesic effect on mice, as assayed by using a twisting test model. This effect of BmK IT-AP well characterized at molecular level is first reported among the known scorpion insect neurotoxins.