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Chinese Medical Sciences Journal 1994年第4期9卷 203-208页

作者：肖成祖黄子才张正光叶建新高丽华郭智霞程度胜周鹤山孔惟惟 InstituteofBiotechnology AcademyofMilitaryMedicalSciencesBeijing100071 instituteofmicrobiologyandepidemiology

CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture *** successfully cultivated CD-1 cells to a very high density (over1×107cells/ml). Prourokinase was stably secreted at about 180 IU/ 1e6 cells/24 h. Experiments showed that CD-1 cells growing on Biosilon microcarriers were able to spontaneously release from the microcarriers, then reatthch and proliferate on fresh microcarriers. This makes it very easy to scale up preduction. The microcarriers could be reused several times without affecting adhesion. proliferation and prourokinase secretion. With CMPECC membrane radial flow chromatography and MPG chromatography, the prourokinase in conditioned medium could be purified to a specific activity of 1×105 IU/mg of protein. The purification factor was about 600 fold, and approxiamately 90 % of the biological activity was recovered.

关键词： Biosilon microcarrier CD-1 cells line prourokinase high density cultivation

Development of a New Sampling Medium for Bioaerosols

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Biomedical and Environmental Sciences 2005年第2期18卷 82-86页

作者： JUN-HUIZHAI MEI-LINGCHEN XIU-ZHIXU ZHEN-HAISUN YUZHOU FENG-XIANGCHE RUI-FUYANG instituteofmicrobiologyandepidemiology 20DongdajieFengtaiDistrictBeijing100071China

Objective To develop a new sampling medium for detecting of bioaerosols. Methods The sampling media were tested by using Escherichia coli, Staphylococcus aureus and Serratia marcescens under static and active conditions, preliminary applications were performed using AGI-10 and high volume sampler. Results The average recovery rates were raised to 24.7%, 58.2%, 40.5%, 44.1%, 20.5%, and 15.4%, respectively in six consecutive experiments under static condition for 60 min at room temperature. Four kinds of sampling media were singled out after static experiments, which were referred to as “samplutions” PD1, PX2, TD1, and TX2, respectively. Under the active condition, the protective efficacy of PD1, PX2, TD1, and TX2 was 226% (153/47), 553% (111/17), 150% (120/48), and 268% (419/114), respectively. Conclusion The samplutions have some effects on the subsequent nucleic acid detection, which could be avoided by employing standard nucleic acid extraction procedure. The newly developed samplution can be applied to the detection of bioaerosols.

关键词： Sampling medium Bioaerosols Airborne microorganisms

Modulation of human enteric epithelial barrier and ion transport function by Peyer's patch lymphocytes

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World Journal of Gastroenterology 2004年第11期10卷 1594-1599页

作者： JieChen Lai-LingTsang Lok-SzeHo DewiK.Rowlands Jie-YingGao Chuen-PeiNg Yiu-WaChung Hsiao-ChangChan EpithelialCellsBiologyResearchCenter DepartmentofPhysiologyFacultyofMedicineChineseUniversityofHongKongHongKongChina DepartmentofImmunology InstituteofMicrobiologyandEpidemiologyAcademyofMilitaryMedicalSciencesBeijing100071China

AIM: To investigate the role of Peyer's patch lymphocytes in the regulation of enteric epithelial barrier and ion transport function in homeostasis and host defense. METHODS: Mouse Peyer's patch lymphocytes were co-cultured with human intestinal epithelial cell line Caco-2 either in the mixed or separated (isolated but permeable compartments) culture configuration. Barrier and transport functions of the Caco-2 epithelial monolayers were measured with short-circuit current (Isc) technique. Release of cytokines was measured by enzyme-linked immunosorbent assay (ELISA) and cytokine mRNA expression was analyzed by semi-quantitative RT-PCR. Barrier and iontransport functions of both culture conditions following exposure to Shigella lipopolysaccharide (LPS) were also examined. RESULTS: The transepithelial resistance (TER) of the epithelial monolayers co-cultured with Peyer's patch lymphocytes was maintained whereas that of the Caco-2 monolayers alone significantly decreased after eight days in culture. The forskolin-induced anion secretion, in either absence or presence of LPS, was significantly suppressed in the both co-cultures as compared with the Caco-2 cells alone. Furthermore, only the mixed co-culture condition induced the expression and release of mIL-6 from Peyer's patch lymphocytes, which could be further enhanced by LPS. However, both co-culture conditions suppressed expression and release of epithelial hIL-8 under the unstimulated conditions, while the treatment with LPS stimulated their hIL-8 expression and release. CONCLUSION: Peyer's patch lymphocytes may modulate intestinal epithelial barrier and ion transport function in homeostasis and host defense via cell-cell contact and cytokine signaling.

关键词：调剂肠上皮通道离子传送功能淋巴细胞修补作用 ELISA 酶联免疫吸附试验

Reovirus,isolated from SARS patients

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Chinese Science Bulletin 2003年第13期48卷 1293-1296页

作者： DUANQing ZHUHong YANGYi LIWeihua ZHOUYusen HEJun HEKun ZHANGHaojie ZHOUTao SONGLihua GANYonghua TANHua JINBaofeng LIHuiyan ZUOTingting CHENDehui ZHANGXuemin instituteofmicrobiologyandepidemiology AcademyofMilitaryMedicalSciencesBeijing100071China ationalCenterofBiomedicalAnalysis Beijing100850China

Beijing has been severely affected by SARS, and SARS-associated coronavirus has been confirmed as its cause. However, clinical and experimental evidence implicates the possibility of co-infection. In this report, reovirus was isolated from throat swabs of SARS patients, including the first case in Beijing and her mother. Identification with the electron microscopy revealed the characteristic features of reovirus. 24 of 38 samples from other SARS cases were found to have serologic responses to the reovirus. Primers designed for reovirus have amplified several fragments of DNA, one of which was sequenced (S2 gene fragment), which indicates it as a unique reovirus (orthoreovirus). Preliminary animal experiment showed that inoculation of the reovirus in mice caused death with atypical pneumonia. Nevertheless, the association of reovirus with SARS outbreak requires to be further investigated.

关键词：呼吸道肠道病毒非典型肺炎 SARS 传染病防治电子显微镜

STUDY ON AEROSOL TRANSMISSION ROUTES OF HEMORRHAGIC FEVER WITH RENAL SYNDROME (HFRS)

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China Particuology 2003年第2期1卷 80-83页

作者： FengxiangChe LingyingMeng JianchunLu JinsongLi JunbaoLi MinxiaLiu Authortowhomcorrespondenceshouldbeaddressed. instituteofmicrobiologyandepidemiology AcademyofMilitaryMedicalSciencesBeijing100071P.R.China

The present paper reviews our studies on transmission routes and prophylactic measures of Hemorrhagic Fever with Renal Syndrome (HFRS), including aerosol inhalation, skin injury/mucosa, insect vectors, peroral infection and vertical transmission. The results show that HFRS is transmitted by Hantaan virus via multi-routes. One of them, the aerosol transmission, is perhaps the main route transmitting HFRS from mouse to human beings. The injury and mu-cosa is the main transmission route from mouse to mouse and also an important agent from mouse to human beings. The peroral infection may occur in very serious pollution of foods. The insect vectors may play an important role in the focus of HFRS and in the transmission of HFRS from mouse to human. The epidemiological significance of vertical transmission is yet uncertain. According to the results, four proposals should be emphasized including killing insects and mice, long lasting and combined prophylactics, intensive studies on prophylactic measures on viral aerosols and the blocking of transmission of HFRS from mouse to human via aerosols.

关键词： Hemorrhagic Fever with Renal Syndrome (HFRS) transmission route aerosol

A Genome Sequence of Novel SARS-CoV Isolates: the Genotype, GD-Ins29, Leads to a Hypothesis of Viral Transmission in South China

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Genomics, Proteomics & Bioinformatics 2003年第2期1卷 101-107页

作者： E‘deQin XiongleiHe WeiTian YongLiu WeiLi JieWen BingyinSi YongwuHu WenmingPeng LinTaug TaoJiang JianpingShi JiaJia YuZhang JiaYe Cui’eWang YujunHan JingqiangWang BaochangFan QingfaWu GuohuiChang WuchunCao ZuyuanXu RuifuYang JmgWang ManYu YanLi JingXu JunZhou YajumDeng XiaoyuLi JianfeiHu CaipingWang ChunxiaYan QingrunZhang JingyueBao GuoqingLi HaiqingZhang YilinZhang HuiZhao XiaoweiZhang ShuangliLi XiaoJieCheng XiuqingZhang BinLiu ChangqingZeng HuanmingYang WeijunChen LinFang ChangfengLi MengLei DaweiLi WeiTong XiangjunTian JianWang BoZhang SonggangLi XuehaiTan SiqiLiu WeiDong JunWang GaneKa-ShuWong JunYu QingyuZhu instituteofmicrobiologyandepidemiology ChineseAcademyofMilitaryMedicalSciencesBeijing100071China BeijingGenomicsInstitute ChineseAcademyofSciencesBeijing101300JamesD.WatsonInstituteofGenomeSciencesZhijiangCampusZhejiangUniversityHangzhou310008China

We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral genome replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.

关键词： Severe Acute Respiratory Syndrome (SARS) genotype GD-Ins29

Preparation, characterizationand preliminary in vivo stud-ies of inactivated SARS-CoV vaccine

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Chinese Science Bulletin 2003年第23期48卷 2621-2625页

作者： TANGLin WANGJian QINEde ZHUQingyu YUMan DINGZhifen SHIHuiying CHENGXiaojie WANGCaiping CHANGGuohui LIShuangli ZHANGXumin CHENXishu YUJun CHENZe BeijingGenomicsInstitute ChineseAcademyofSciencesBeijing101300China instituteofmicrobiologyandepidemiology AcademyofMilitaryMedicineBeijing100071China NationalVaccineandSerumInstitute Beijing100024China WuhanInstituteofVirology ChineseAcademyofSciencesWuhan430071China

A large quantity of SARS-CoV virus was pro-liferated in Vero cells, inactivated with b-propiolactone, then purified by Sepharose 4FF column chromatography to pre-pare inactivated vaccine. The vaccine was identified by Western blot, mass spectrographic analysis, ELISA and elec-tron microscopy. The vaccine with or without aluminum hydroxide adjuvant was inoculated into female BALB/c mice at different dosages. The result showed that the antibodies to SARS-CoV were induced in the mice. The antibody levels induced by the vaccine with aluminum hydroxide were higher than those without aluminum hydroxide.

关键词：疫苗非典型肺炎 SARS β-丙内酯中国

Sequence evolution of putative cytotoxic T cell epitopes in NS3 region of hepatitis C virus

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World Journal of Gastroenterology 2004年第6期10卷 847-851页

作者： Hua-ZhangGuo YingYin Wen-LiangWang Chuan-ShanZhang TaoWang ZheWang JingZhang HongCheng Hai-TaoWang DepartmentofPathology XijingHospitalFourthMilitaryMedicalUniversityXi'an710033ShaanxiProvinceChina DepartmentofClinicalLaboratories XijingHospitalFourthMilitaryMedicalUniversityXi'an710033ShaanxiProvinceChina instituteofmicrobiologyandepidemiology AcademyofMilitaryMedicalSciencesBeijing100071China DepartmentofPathology XijingHospitalFourthMilitaryMedicalUniversityXi'an710033ShaanxiProvinceChina

AIM:Quasispecies of hepatitis C virus (HCV) are the foundation for rapid sequence evolution of HCV to evade immune surveillance of hosts. The consensus sequence evolution of a segment of HCV NS3 region, which encompasses putative cytotoxic T cell epitopes,was ***:Three male patients, infected with HCV through multiple transfusions,were identified from clinical symptoms and monitored by aminotransferase for 60 *** samples taken at months 0, 32, and 60 were used for viral RNA extraction. A segment of HCV NS3 region was amplified from the RNA extraction by RT-PCR and subjected to subcloning and *** types of these three patients were determined using complement-dependent microlymphocytotoxic assay. CTL epitopes were predicted using MHC binding ***:No patient had clinical symptoms or elevation of aspartate/alanine arninotransferase. Two patients showed positive HCV PCR results at all 3 time points. The other one showed a positive HCV PCR result only at month 0.A reported HLA-A2-restricted CTL epitope had no alteration in the HLA-A2-negative carrier over 60 *** the HLA-A2-positive individuals, all the sequences from 0 month 0 showed an amber mutation on the initial codon of the epitope. Most changes of consensus sequences in the same patient occurred on predicted cytotoxic T cell ***:Amber mutation and changes of consensus sequence in HCV NS3 region may be related to viral immune escape.

关键词：丙型肝炎病毒细胞毒素 T细胞抗原决定基 NS3区逆转录聚合酶链反应

QUANTITATIVE STUDY ON THE EXPRESSION OF MMP1 AND TIMP1 IN NORMAL AND RADIATION-COMBINED WOUND HEALING AND ITS EFFECTS ON THE HEALING PROCESS

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中国体视学与图像分析 2002年第1期7卷 15-19,57页

作者： GUQing-yang ZHOUJie InstituteofRadiationMedicine AcademyofMilitaryMedicalSciencesBeijing100850China instituteofmicrobiologyandepidemiology AcademyofMilitaryMedicalSciencesBeijing100850China

Aim: To study the expression of MMP1 and TIMP1 in normal and radiation-combined wound healing and their effects on the healing process. Materials and Methods: A rat model of radiation-combined wound healing was used, and routine light microscopy, electron microscopy, immunohistochemistry, and in situ hybridization were used to detect the expression of MMP1 and TIMP1 during the healing process. Results: The wound healing process was impaired and delayed. In rats receiving 25Gy Gamma ray locally, the irradiated wounds healed 6 days later than non-irradiated controls. The following changes were found in the expression of MMP1 and TIMP1: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression was only slightly if at all affected in the newly-formed epidermis of irradiated wounds when compared with that in controls. Later, the epidermal expression of MMP1 in irradiated wounds was comparatively increased following the delayed healing process. 3 to 14 days after wounding, TIMP1 was weakly positive in proliferating keratinocytes of control group and became negative after epidermal covering, whereas no or only slight epidermal expression was detected in the irradiated group before epidermal covering. (2) The expression of MMP1 and TIMP1 in irradiated group was markedly decreased in fibroblasts, endotheliocytes and macrophages when compared to that in controls. The expression phase was prolonged due to the delayed healing process. Conclusions: It is concluded that the reduced expression of MMP1 and TIMP1 in granulation tissue retards such important processes as cell migration and angiogenesis, thus slowing the healing process. The expression of MMP1 in the newly-formed epidermis may help the process of reepithelialization, but in the late healing period, overexpression of MMP1 and decreased expression of TIMP1 in the epidermis may hinder the establishment of basal membrane and scar formation.

关键词：放射损伤愈合 MMP1 TIMP1 动物实验显微观察免疫组织化学

Complete Genome Sequences of the SARS-CoV: the BJ Group (Isolates BJ01-BJ04)

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Genomics, Proteomics & Bioinformatics 2003年第3期1卷 180-192页

作者： ShengliBi E‘deQin ZuyuanXu WeiLi JingWang YongWuHu YongLiu ShuminDuan JianfeiHu YujunHan JingXu YanLi YaoYi YongdongZhou WeiLin1 JieWen HongXu RuanLi ZizhangZhang HaiyanSun JinguiZhu ManYu BaochangFan QingfaWu WeiLin2 LinTang Bao’anYang GuoqingLi WenmingPeng WenjieLi TaoJiang YajunDeng BohuaLiu JianpingShi YongqiangDeng WeiWei HongLiu ZongzhongTong FengZhang YuZhang Cui‘eWang YuquanLi JiaYe YonghuaGan JiaJi XiaoyuLi XiangjunTian FushuangLu GangTan RuifuYang BinLiu SiqiLiu SonggangLi JunWang JianWang WuchunCao JunYu XiaopingDong HuanmingYang CenterofDiseaseControlandPrevention Beijing100050China instituteofmicrobiologyandepidemiology ChineseAcademyofMilitaryMedicalSciencesBeijing100071China JamesD.WatsonInstituteofGenomeSciences ZhijiangCampusZhejiangUniversityandHangzhouGenomicsInstituteHangzhou310008China BeijingGenomicsInstitute ChineseAcademyofSciencesBeijing101300China CollegeofLifeSciences PekingUniversityBeijing100871China WenzhouMedicalCollege Wenzhou325003China PekinaUnionMedicalColleqeHospital ChineseAcademyofMedicalSciencesBeijing100730China CollegeofMaterialsScienceandChemicalEngineering YuquanCampusZhejiangUniversityHangzhou310027China

Beijing has been one of the epicenters attacked most severely by the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) since the first patient was diagnosed in one of the city's hospitals. We now report complete genome sequences of the BJ Group, including four isolates (Isolates BJ01, BJ02, BJ03, and BJ04) of the *** is remarkable that all members of the BJ Group share a common haplotype, consisting of seven loci that differentiate the group from other isolates published to date. Among 42 substitutions uniquely identified from the BJ group, 32 are non-synonymous changes at the amino acid level. Rooted phylogenetic trees, proposed on the basis of haplotypes and other sequence variations of SARS-CoV isolates from Canada, USA, Singapore, and China, gave rise to different paradigms but positioned the BJ Group, together with the newly discovered GD01 (GD-Ins29) in the same clade, followed by the H-U Group (from Hong Kong to USA) and the H-T Group (from Hong Kong to Toronto), leaving the SP Group (Singapore) more distant. This result appears to suggest a possible transmission path from Guangdong to Beijing/Hong Kong, then to other countries and regions.

关键词： SARS SARS-CoV haplotype substitution phylogeny