Extraction and purification of TGFβ and its effect on the induction of apoptosis of hepatocytes

AIM To extract and purify the transforming growth factor β (TGF β), and to demonstrate its biological activity in vivo and induction of apoptosis of hepatocytes in vitro.METHODS TGF β was isolated from fresh bovine platelets by acid/ethanol extraction method and purified with ion exchange and gel chromatography. The extracted TGF β was injected subcutaneously to mice, and its biological activity in vivo was observed 72 hfs post-injection by HE staining. The morphological changes were observed by HE staining and the occurrence of apoptosis was detected by TUNEL method after the human normal hepatic cell line QZG was treated with 8 μg@L 1 TGFβ for 12 hrs in vitro.RESULTS The molecular mass 25 ku TGF β protein was successfully extracted. It was able to induce localized granulation tissue formation in vivo. TGF β-treated hepatocytes showed obvious apoptotic morphological changes, including the pyknosis and dense-stained nuclei and cytoplasm, the fragmentary, annular or crescent nuclei, and the "bubbling" cytoplasm. Moreover, its apoptotic rate was significantly higher than that of the control group (P＜0.05).CONCLUSION Biological active TGF β protein is extracted and purified successfully from bovine platelets, and it is able to induce the apoptosis of hepatocytes.

Design a pediatric isolation bed and characterization of its purification efficacy in ambient air

Patients with acute infectious respiratory illness may emit bio‐aerosols containing pathogens to infect susceptible hosts,including other patients and healthcare workers.This study aimed to design the pediatric isolation(PI)bed and evaluate its purification effect on aerosols and microorganisms in the air in experiment rooms and verify the general wards'purification efficiency.The PI bed is connected to a purification device that surrounds the child's head to prevent contaminants'spread into the room.We test the purification effect of the PI bed in the experiment room and verified the purification effect of the PI bed in the hospital.The results showed that PI beds in the demonstration ward presented purifying effects on both smoke particulates and microorganisms like Staphylococcus albus(S.albus)and H1N1 virus.The Aerosol in the room in the hospital was then detected to validity the purification rate.Therefore,Isolation beds could be used in hospitals to reduce the risk of nosocomial infection to protect the health of doctors,patients,and visitors.

栉孔扇贝一种球形病毒的分离纯化及其超微结构观察

Electron microscopic examinations of ultrathin sections reveal the presence of spherical virus like particles in the digestive gland, kidney, and the intestine of moribund scallop Chlamys farreri collected during a severe mortality in Jiaozhou Bay (Qingdao, China) in 2000. The virions are approximately 130 to 170 nm in diameter and have a bilaminal envelope, while the nucleocapsids are 90-140 nm. The virus occurs and distributes in the cytoplasmic vesicles of infected cells, and it is arranged in a scattered form without occluding protein. The viral particles are consistent in size and morphology in preparations derived from the isolation and also from ultrathin sections. The spikes of envelope are clearly observed with negative staining. By the same methods, this virus was not found in the healthy scallop, indicating that this virus might be the causative aetiology for the abnormal mortality.

兔轮状病毒的分离与鉴定

目的 从疑似病料中分离免轮状病毒。方法 从形态学、血清学、分子生物学等几个方面对其进行鉴定。结果 从几个方面鉴定的结果显示 ,疑似病料中含有兔轮状病毒 (LaRV)病毒粒子。结论 本次实验成功地分离得到了LaRV病毒。

提纯CVB1的方法学改进和CVB1的灭活

目的：研究聚乙二醇沉淀和蔗糖梯度离心法提纯柯萨奇病毒Ｂ１（ＣＶＢ１） 的效率，并采用β丙内酯（ＢＰＬ） 将其灭活，观察ＢＰＬ的灭活效果。方法：①聚乙二醇沉淀法将病毒悬液初步浓缩；②在１０％ ～５０％ （ ｍ ／ Ｖ） 蔗糖梯度中上样，低温超速离心；③电镜观察并做病毒活性测定；④ＢＰＬ对病毒进行灭活，测定残存毒力。结果：①病毒在５０％ （ ｍ ／ Ｖ） 蔗糖处有一吸收峰，并被电镜证实；②ＢＰＬ灭活病毒彻底，灭活后病毒稀释１０２ ～１０１０ 倍接种细胞未见病变。结论：ＣＶＢ１ 经聚乙二醇沉淀和蔗糖梯度超速离心后收获率高，ＢＰＬ灭活彻底，保证了进一步实验的成功。

Preparation and antiviral effects of polysaccharides from Panax japonicus

Water-soluble polysaccharides were prepared from Panax japonicus by hot water extraction and ethanol precipitation.The polysaccharides were further purified by ion exchange chromatography to obtain neutral and acidic polysaccharides.The neutral polysaccharide fraction mainly contained Glc(90.2%),which was a glucan fraction.The acidic polysaccharide fraction mainly contained GalA(43.6%),Gal(21.7%),and Ara(15.4%),with a degree of methyl-esterification of 20.3%,which was a pectic polysaccharide.The acidic polysaccharide of Panax japonicus could effectively inhibit the replication of human seasonal influenza virus H1N1 and canine influenza virus H3N2 in MDCK cells and A549 cells and significantly reduce the virus titer in infected cells.It also effectively inhibited the number of infected cells of the SARS-CoV-2 South Africa strain and the Omicron strain.The acid polysaccharide of Panax japonicus showed good efficacy against influenza virus and COVID-19 infection,which could be used as a potential antiviral candidate drug molecule in the future.

Studies of Coltivirus in China

Objective The purpose of this article is to review the developments of studies of Coltivirus in ChinaData sources The data used in this review was obtained mainly from the studies of Coltivirus reported from 1990 to 2003 in China.Study selection Relevant articles on studies of Coltivirus in domestic and foreign literature were selected.Data extraction Data were maily extracted from the articles which are listed in the reference section of this review.Results Many Coltiviruses have been isolated not only from blood samples of patients with unknown fever or from cerebrospinal fluid of patients with encephalitis in Xishuangbanna area in Yunnan province, but also from mosquitoes collected in many areas in China. In some patients diagnosed as Japanese encephalitis or unknown fever, an increase of Coltivirus IgG antibody of fourfold, or more, has been detected using ELISA. Similarly, Coltivirus IgM antibody was positive in some patients with Japanese encephalitis or viral encephalitis. From most Chinese patients, except the northeastern, the isolates of Coltiviruses belong to subgroup B2, according to RT-PCR amplification of the ninth and twelfth segments of the isolates and sequence analysis of their amplicons. Some biological properties of Chinese Coltiviruses isolates are different from that of North American Coltiviruses.Conclusions The isolates of Coltiviruses from Chinese patients are one of the common agents causing viral encephalitis and unknown fever in summer-autumn season. It might be an important public health problem due to its high isolation rate and wide distribution in China. Mosquito is the main transmission vector of the virus.

河北省虫媒病毒分离鉴定

目的 研究河北省蚊虫携带虫媒病毒情况。方法 在蚊虫孳生地采集标本液氯保存，按照20—30只／组经无菌处理后研磨，研磨液离心后接种C6／36细胞，连续三代观察致细胞病变情况；对细胞病变阳性的分离物，进行血清学和分子生物学鉴定。结果 在河北省涉县两个村共采集蚊虫1310只，分46组进行处理，共获得13株阳性分离物，其中两株分离物经间接免疫荧光检测提示为甲病毒属成员，用甲病毒属特异性引物RT-PCR扩增阳性，经核酸序列测定证实该序列与Getah病毒（AY702913．1）同源性最高（98％），初步鉴定为Getah病毒，其他病毒分离物尚在鉴定。结论 本研究从河北省蚊虫标本中分离到Getah病毒，这是我国内陆地区首次分离到该病毒。

慢性丙型肝炎患者血清和肝组织中庚型肝炎病毒RNA检测

目的了解慢性丙型肝炎（ＣＨＣ）患者血清和肝组织中庚型肝炎病毒（ＨＧＶ）感染的状况．方法用ＲＴＰＣＲ方法对４９例ＣＨＣ患者血清和其中１４例肝组织进行ＨＧＶＲＮＡ检测．结果在４９例ＣＨＣ患者的血清和其中１４例肝组织中ＨＧＶＲＮＡ阳性检出率分别为１２２％和２１４％，证实ＨＧＶ和ＨＣＶ感染在ＣＨＣ患者血清和肝组织中同时存在，而肝组织中ＨＧＶＲＮＡ阳性检出率更高，其原因同受血或其直接经血液暴露密切相关．结论ＣＨＣ患者血清和肝组织中存在ＨＧＶ感染，而且感染率较高，故应加强对献血员进行ＨＧＶＲＮＡ和ＡＬＴ筛查。

PCR-RDB技术快速检测儿童急性呼吸道感染DNA病毒的研究

目的建立多重PCR反向斑点杂交方法（PCR-RDB）技术快速检测儿童急性呼吸道感染DNA病毒，以便及时监测并快速诊断急性病毒性呼吸道感染。方法参照国家生物技术信息中心（NCBI）数据库病毒核酸序列设计多重PCR引物和探针，将特异的寡核苷酸探针固定于尼龙膜上。利用多重PCR对人博卡病毒（hBOV）、KI多瘤病毒（KI）、腺病毒（AdV）、WU多瘤病毒（WUPyV）、人细小病毒B19（HPVB19）等5种DNA病毒进行扩增，扩增产物变性后与各特异性探针进行杂交、显色并分析结果，并对其敏感性、特异性进行测试。同时通过反向斑点杂交检测108例临床标本的多重PCR产物，并与培养法结果做对比分析。结果建立的多重PCR—RDB方法各特异探针只相应扩增产物杂交，与其他病原体无交叉反应，敏感性达1 CFU。108例临床咽拭子标本PCR—RDB方法共筛选出阳性37例（34．26％），高于临床常规检测方法的30例（27．78％）。结论应用多重PCR-RDB技术可简便、快速、准确地检测儿童急性呼吸道感染DNA病毒，具有很好的临床应用前景。

极端嗜盐多形性古菌病毒HRPV13的分离鉴定及生物学特性

【背景】与感染细菌和真核生物的病毒相比,目前发现的古菌病毒数量很少,但是却展现出形态多样性。因此,分离和鉴定新的古菌病毒具有重要意义。【目的】为了进一步了解古菌病毒的多样性,我们从青海省翡翠湖水样中分离到一株新的嗜盐古菌病毒株,研究其生物学特性并进行分类。【方法】首先通过挑取单菌落法分离嗜盐古菌,通过噬菌斑法获得嗜盐古菌病毒,PEG 6000两步沉淀法和CsCl密度梯度离心对病毒颗粒进行浓缩和纯化,用醋酸双氧铀对病毒负染染色,在透射电镜下观察病毒形态,提取病毒基因组后进行测序并进行生物信息学分析,以三氯乙酸法制备病毒蛋白样品并进行SDS-PAGE分析,分别用考马斯亮蓝和苏丹黑B染色并观察其蛋白和脂质条带。【结果】在以Halorubrum属极端嗜盐古菌K2菌株为敏感菌的双层平板上分离到了一株嗜盐古菌病毒,其噬菌斑为浊斑,透射电镜下呈多形性包膜病毒状,直径60 nm左右;含有9333 bp大小的双链环状DNA基因组,与已报道的β多形包膜病毒属(Betapleolipovirus)的HRPV11、HRPV12和HRPV10具有约75%的一致性,是该属的一个病毒新种。根据形态及基因组特征,将其归属于β多形包膜病毒属,命名为Halorubrum pleomorphic virus 13(HRPV13)。该病毒在50℃以下及pH 5.0−9.0具有较好活性,高盐浓度下具有侵染能力,而且病毒侵染宿主后不会造成宿主的大量裂解。【结论】通过电镜观察和生物学特性研究以及基因组测序分析,证实HRPV13为β多形包膜病毒属的新种,其分离和鉴定丰富了多形性病毒种类,这为研究不同区域多形性病毒的进化奠定了基础。