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Study on the determinants of suckling mice neurovirulence of dengue 2 virus

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Science China(Life Sciences) 2003年第1期46卷 95-103页

作者：赵卫范宝昌胡志君陈水平王鹏程苑锡同李晓萸于曼秦鄂德杨佩英 institute of Microbiology & epidemiology Academy of Military Medical Sciences

pDVWS501 was a genomic-length cDNA clone of dengue 2 virus, through which infec-tious virus (MON501) could be rescued. MON501 was neurovirulent in mice, whose E residues 62 and 203 were Lys and Asn, respectively. Two genomic-length cDNA clones (TB62 and TB203) were constructed by pointed mutation of pDVWS501 with OL-PCR, E62 of TB62 and E203 of TB203 were converted to Glu and Asp, respectively. RNA transcripts of pDVWS501, TB62 and TB203 were produced in vitro and electroporated into BHK-21 cells. The cultures were collected after 7 days and used as inoculum to infect C6/36 cells. The existence of rescued dengue viruses in the culture was proved by RT-PCR, and the typical cytopathic effect (CPE) of C6/36 caused by dengue virus emerged after 2—5 days?inoculation. Sequence analysis further confirmed the exis-tence of recovered and recombinant DEN2 viruses, whose 5′ termini had an additional non-virus nucleotides 揋? while the 3′ terminal sequences remained the same as natural. The neuroviru-lence of three viruses was evaluated in 1-day-old mice by the intracerebral route with 105—102 TCID50. Compared with MON501 group, the number of infected mice with the signs of encephalitis in HFT62 and HFT203 groups was less, and the surviving time was longer. The properties of these mutants demonstrated that E62 and E203 are determinants of suckling mice neurovirulence.

关键词： dengue 2 virus, virulent determinants, infectious transcripts, point mutation.

Development of Cell Lines Stably Expressing Staphylococcal Nuclease Fused to Dengue 2 Virus Capsid Protein for CTVI

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 577-582页

作者： Cheng-Feng QIN and E-De QIN institute of Microbiology and epidemiology, Academy of Military medical Sciences, Beijing 100071, China institute of Microbiology and epidemiology Academy of Military Medical SciencesBeijing 100071 China

To explore the potential application of capsid-targeted viral inactivation (CTVI) strategy in prophylactic model against dengue virus (DV) infection, here we fused a Ca2+-dependent nuclease, staphylococcal nuclease (SN), to the capsid protein of dengue 2 virus (D2C) at the carboxyl terminal, and constructed the desired expression plasmid pc/D2C-SN and control plasmids pc/D2C-SN* and pc/D2C. A mammalian cell line BHK-21 was transfected by electroporation with those plasmids and thereafter selected by 5 μg/ml blasticidin. The resistant cell clones were then expanding cultured and screened by RT-PCR and Western Blot assays. The nuclease activity of the expressed fusion protein D2C-SN was analyzed by in vitro DNA digestion assay. It was confirmed cell lines stably expressing D2C-SN and control constructs were obtained. The intracellular expressed fusion protein D2C-SN had ideal nuclease activity and no cytotoxicity on mammalian cells. Those engineered cell lines provided the experimental system for CTVI application in prophylactic model and paved the new road for combating DV infection with CTVI.

关键词： dengue 2 virus capsid protein staphylococcal nuclease CTVI stable expression

Assessment of quality of life for the patients with cervical cancer at different clinical stages

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Chinese Journal of Cancer 2013年第5期32卷 275-282页

作者： Yao Xie Fang-Hui Zhao Si-Han Lu He Huang Xiong-Fei Pan Chun-Xia Yang You-Lin Qiao Department of epidemiology West China Public Health School Sichuan University Department of epidemiology Cancer Institute/Hospital Chinese Academy of Medical Sciences

With improved overall survival of cervical cancer patients, the importance of the quality of life (QOL) is increasingly recognized. This study was conducted to compare the QOL of women with different stage cervical cancer before and after treatment to facilitate improved cervical cancer prevention and treatment. We used the generic medical Outcomes Study Short Form-36 (MOS SF-36) to collect QOL information. Based on SF-36, we interviewed cervical cancer patients at West China Second Affiliated Hospital and Sichuan Cancer Hospital between May 2010 and January 2011. A total of 92 patients with precancerous lesions, 93 with early cancer, and 35 with advanced cancer responded to our survey. Average physical component summary (PCS) scores were significantly different between the three groups at every time point (P < 0.05). Average mental component summary (MCS) scores were significantly different between the three groups after treatment (P < 0.05). Average PCS and MCS scores increased gradually from the pretreatment to posttreatment period for patients with precancerous lesions. However, they reached the lowest at 1 month after treatment for patients with early and advanced cancers and rebounded between 1 and 6 months after treatment. Our results indicate that patients with precancerous lesions and early cervical cancer show better overall QOL than do those with advanced cervical cancer. Additionally, patients with early cancer recover more quickly than do those with advanced cancer in terms of both physical and mental functions. Thus, early detection and treatment initiatives may improve the QOL for patients with precancerous lesions and cervical cancer.

关键词：子宫颈癌癌症患者生活质量质量评估临床肿瘤医院中国西部精神功能

Microalbuminuria in hepatitis C-genotype 4:Effect of pegylated interferon and ribavirin

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World Journal of Gastroenterology 2010年第10期16卷 1226-1231页

作者： Moutaz Derbala Fatma M Shebl Awad Rashid Aliaa Amer Abdulbari Bener Department of Gastroenterology and Hepa-tology Hamad Medical Corporation Department of Medicine Weill Cornell Medical College in Qatar Infections & Immunoepidemiology Branch Division of Cancer Epidemiology and GeneticsNational Cancer Institute epidemiology Branch National Liver Institute Department of Nephrology Hamad Medical Corporation Department of Laboratory Medicine and Histo-pathology Hematology SectionHamad Medical Corporation Department of medical Statistics and epidemiology Hamad Medical Corporation

AIM:To study the relation between hepatitis C virus (HCV) genotype 4 and microalbuminuria and renal impairment in relation to hepatic histology, and viremia in the absence of cryoglobulinemia, and to examine the effect of treatment on ***: Three hundred subjects, including 233 HCV genotype-4 infected patients, were tested for cryoglobulinemia, microalbuminuria, albumin creatinine ratio (ACR), urea, creatinine, and estimated glomerular filtration rate (eGFR). The parameters were measured again in the HCV patients after 48 wk of treatment with pegylated interferon and ***: Significantly higher levels of microalbumin- uria were detected in HCV-positive patients compared to HCV-negative controls (median 9.5 vs 5.9, respectively, Kruskal-Wallis P=0.017). Log microalbuminuria was significantly correlated with hepatic inflammation (r=0.13, P=0.036) and f ibrosis (r=0.12, P=0.061), but not with viral load (r=-0.03, P=0.610), or alanine transaminase (r=-0.03, P=0.617). Diabetes mellitus neither significantly moderated (χ2=0.13, P=0.720), nor mediated (Sobel test P=0.49) the HCV effect. HCV status was signifi cantly associated with log microalbu-minuria (χ2=4.97, P=0.026), adjusting for age, gender, diabetes, cryoglobulinemia, urea and creatinine. A positive HCV status was not significantly associated with low eGFR (<60 mL/min every 1.73 m2) [odds ratio (OR): 0.5, 95% confidence interval (CI):0.2-1.4], nor with high ACR (OR: 1.7, 95% CI:0.7-4.1). End-of-treatment response (ETR) was achieved in 51.9% of patients. Individuals with ETR had significantly lower microalbuminuria post-treatment (χ2=8.19, P=0.004).CONCLUSION: HCV affected the development of microalbuminuria independent of diabetes or cryoglobulinemia. Combination therapy of pegylated interferon-ribavirin had a positive effect in reducing microalbuminuria.

关键词： Hepatitis C virus Genotype Kidney diseases Albuminuria Proteinuria Peginterferon α-2a Ribavirin

Direct cloning and transplanting of large DNA fragments from Escherichia coli chromosome

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Science China(Life Sciences) 2016年第10期59卷 1034-1041页

作者： Ying Zhu Yan Yang Pingping Den Yong Huang Mengxiang Ni Hongqing Fang institute of Biotechnology Academy of Military Medical Sciences institute of Life Science and technology China Pharmaceutical University institute of Health Sciences Anhui University institute of Microbiology and epidemiology Academy of Military Medical Sciences

We applied a resistance split-fusion strategy to increase the in vivo direct cloning efficiency mediated by Red recombination. The cat cassette was divided into two parts： cma （which has a homologous sequence with cmb） and cmb, each of which has no resistance separately unless the two parts are fused together. The crab sequence was integrated into one flank of a target clon- ing region in the chromosome, and a linear vector containing the cma sequence was electroporated into the cells to directly capture the target region. Based on this strategy, we successfully cloned an approximately 48 kb DNA fragment from the E. coli DH1-Z chromosome with a positive frequency of approximately 80%. Combined with double-strand breakage-stimulated homologous recombination, we applied this strategy to successfully replace the corresponding region of the E. coli DH36 chromosome and knock out four non-essential genomic regions in one step. This strategy could provide a powerful tool for the heterologous expression of microbial natural product biosynthetic pathways for genome assembly and for the functional study of DNA sequences dozens of kilobases in length.

关键词： Red homologous recombination resistance split-fusion target cloning transferring

Risk of second primary cancers after testicular cancer in East and West Germany： a focus on contralateral testicular cancers

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Asian Journal of Andrology 2014年第2期16卷 285-289,I0010,I0011页

作者： Carsten Rusner Brigitte Streller Christa Stegmaier Pietro Trocchi Oliver Kuss Katherine A McGlynn Britton Traberts Andreas Stang institute of Clinical epidemiology Medical Faculty Martin-Luther-University of Halle-Wittenberg Halle (Saale) Common Cancer Registry of Berlin Berlin Saarland Cancer Registry Saarbrucken institute of medical epidemiology Biostatistics and Informatics Medical Faculty Martin-Luther-University of Halle-Wittenberg Halle (Saale) Germany Division of Cancer epidemiology and Genetics National Cancer Institute National Institutes of Health Bethesda MD Department of epidemiology Scfloof of Public Heaftfl Boston University Boston MA USA.

Testicular cancer survival rates improved dramatically after cisplatin-based therapy was introduced in the 1970s. However, chemotherapy and radiation therapy are potentially carcinogenic. The purpose of this study was to estimate the risk of developing second primary cancers including the risk associated with primary histologic type （seminoma and non-seminoma） among testicular cancer survivors in Germany. We identified 16 990 and 1401 cases of testicular cancer in population-based cancer registries of East Germany （1961-1989 and 1996-2008） and Saarland （a federal state in West Germany; 1970-2008）, respectively. We estimated the risk of a second primary cancer using standardized incidence ratios （SIRs） with 95% confidence intervals （95% Cls）. To determine trends, we plotted model-based estimated annual SIRs. In East Germany, a total of 301 second primary cancers of any location were observed between 1961 and 1989 （SIR： 1.9; 95% Ch 1.7-2.1）, and 159 cancers （any location） were observed between 1996 and 2008 （SIR： 1.7; 95% Ch 1.4-2.0）. The SIRs for contralateral testicular cancer were increased in the registries with a range from 6.0 in Saarland to 13.9 in East Germany. The SIR for seminoma, in particular, was higher in East Germany compared to the other registries. We observed constant trends in the model-based SIRs for contralateral testicular cancers. The majority of reported SIRs of other cancer sites including histology-specific risks showed low precisions of estimated effects, likely due to small sample sizes. Testicular cancer patients are at increased risk especially for cancers of the contralateral testis and should receive intensive follow-ups.

关键词： cancer registry incidence neoplasms second primary testicular neoplasms

Cloning and expression of 5,10-Methylenetetrahydrofolate reductase (MTHFR) gene

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Science China(Life Sciences) 1998年第6期41卷 636-643页

作者：陈光慧张晨晖张霄瑜朱燕青汤健孟昭亨李竹 institute of Cardiovascular Research Beijing Medical University Beijing China Department of Health Care epidemiology Beijing Medical University Beijing China

Hyperhomocysteinemia is an independent risk factor of cardiovascular diseases and birth defects. One of the important factors causing hyperhomocysteinemia is decrease of 5,10 Methylenetetrahydrofolate reductase. Human and rat MTHFR cDNAs with RT PCR were isolated, a prokaryodytic expression vector containing human MTHFR cDNA was constructed, and human MTHFR protein was expressed in \%E. coli.\% It was also found that the expression of rat MTHFR could be promoted by IL 1 and homocysteine.

关键词： MTHFR cloning and expression.

MicroRNA-506 is up-regulated in the development of pancreatic ductal adenocarcinoma and is associated with attenuated disease progression

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Chinese Journal of Cancer 2016年第12期35卷 666-672页

作者： Run-Fen Cheng Jian Wang Jing-Yi Zhang Lin Sun Yan-Rui Zhao Zhi-Qiang Qiu Bao-Cun Sun Yan Sun Department of Pathology Tianjin Medical University Cancer Institute and Hospital Department of Pancreatic Oncology Tianjin Medical University Cancer Institute and Hospital Department of epidemiology and Biostatistics Tianjin Medical University Cancer Institute and Hospital Department of Pathology Tianjin Medical University

Background: MicroRNA?506(miR?506) has been reported to function in several tumors as a tumor suppressor gene or oncogene. However, the expression and role of miR?506 in pancreatic ductal adenocarcinoma(PDAC) remains unclear. In this study, we aimed to evaluate the phenotype of miR?506 in ***: Using mi RNA in situ hybridization, we examined the expression of miR?506 in 113 PDACs and 87 paired normal pancreatic tissues. We evaluated mi R?506 expression in PDAC cells, normal pancreatic ducts, and acinus/islands, and we analyzed the associations between miR?506 expression and the clinicopathologic characteristics of PDAC ***: miR?506 expression was higher in PDAC than in matched normal pancreatic ductal cells(P < 0.001). On the other hand, the combined group of well and moderately diferentiated PDACs showed higher levels of mi R?506 than the poorly diferentiated ones(P = 0.023). Moreover, mi R?506 expression was negatively associated with pathologic T category(P = 0.004) and lymph node metastasis(P = 0.033), suggesting that mi R?506 might inhibit the progression of ***: Our results suggest that mi R?506 either plays a role as an oncogene in the tumorigenesis and a tumor suppressor in the progression or serves as a house?keeping, tumor?suppressing mi RNA, whose expression can be activated by oncogenic signals in early development to hinder the progression of PDAC.

关键词： Pancreatic ductal adenocarcinoma miR?506 Tumor suppressor Development Progression

Population-based study of DNA image cytometry as screening method for esophageal cancer

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World Journal of Gastroenterology 2012年第4期18卷 375-382页

作者： Lin Zhao,Wen-Qiang Wei,Xin-Qing Li,Guo-Qing Wang,Qi Shang,You-Lin Qiao,Department of Cancer epidemiology,Cancer institute/Hospital,Chinese Academy of medical Sciences and Peking Union medical College,Beijing 100021,China De-Li Zhao,Department of epidemiology,Feicheng People’s Hospital,Feicheng 271600,Shandong Province,China Chang-Qing Hao,Department of Endoscopy,Linzhou Cancer Hospital,Linzhou 456500,Henan Province,China Dong-Mei Lin,Qin-Jing Pan,Department of Pathology,Cancer institute/Hospital,Chinese Academy of medical Sciences and Peking Union medical College,Beijing 100021,China Fu-Hua Lei,Department of Pathology,Feicheng People’s Hospi-tal,Feicheng 271600,Shandong Province,China Jin-Wu Wang,Department of Pathology,Linzhou Cancer Hospi-tal,Linzhou 456500,Henan Province,China Department of Cancer epidemiology Cancer Institute/Hospital Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100021 China.

AIM:To explore the DNA image cytometry (DNA-ICM) technique as a primary screening method for esopha-geal squamous precancerous ***:This study was designed as a population-based screening study.A total of 582 local residents aged 40 years-69 years were recruited from Linzhou in Henan and Feicheng in ***,only 452 subjects had results of liquid-based cytology,DNA-ICM and *** sensitivity and specificity of DNA-ICM were calculated and compared with liquid-based cytology in moderate dysplasia or ***:Sensitivities of DNA-ICM ranging from at least 1 to 4 aneuploid cells were 90.91%,86.36%,79.55% and 77.27%,respectively,which were better than that of liquid-based cytology (75%).Specifici-ties of DNA-ICM were 70.83%,84.07%,92.65% and 96.81%,but the specificity of liquid-based cytology was 91.91%.The sensitivity and specificity of a combination of liquid-based cytology and DNA-ICM were 84.09% and 85.78%,***:It is possible to use DNA-ICM tech-nique as a primary screening method for esophageal squamous precancerous lesions.

关键词： DNA image cytometry Aneuploidy Screen-ing Esophageal cancer Precancerous lesions