Jianming Gao

Publications (6)7.97 Total impact

• Article: [Identification of EBV chromosomal integration sites in Raji cells by fluorescence in situ hybridization].

  Jianming Gao, Xiaoling Li, Guiyuan Li
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  ABSTRACT: To identify the Epstein-Barr virus (EBV) chromosomal integration sites in Raji cells. EBV DNA was detected by Southern hybridization, and the viral chromosomal integration sites were identified using G banding and fluorescence in situ hybridization (FISH). BamHI-digested genomic DNA from Raji cells was hybridized with (32)P-labeled probe-1 (EBV genome 13,232 approximately 16,189) and Probe-2 (EBV genome 5 approximately 3,271), which generated 4 and 10, 23 kb positive bands respectively. The viral integration sites included 1p, 1q, 2q, 3p, 3q, 4 q, 5q, 6q, 7p, 7q, 9q,11p, 14 q, and 15q,and chromosomal bands 4 q, 2q, 1q and 7q were viral integration sites with high frequencies. Among the 33 signals counted, 7, 4, 4,and 4 signals were at the site 4 q, 2q, 1q, and 7q respectively, and 64% of the total signals were found in these 4 chromosomal bands. No viral integration occurred in chromosomes 16 approximately 22 or the sex chromosomes (X, Y). This study firstly identifies the EBV integration sites in Raji cells using G banding and FISH. There are some viral integration sites with high frequencies in Raji cells, and EBV integrates into Raji cell genomes non-randomly.
  Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 02/2009; 34(1):13-9.
• Article: A case-control study on etiological factors involved in patients with burning mouth syndrome.

  Jianming Gao, Lizhang Chen, Jia Zhou, Jieying Peng
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  ABSTRACT: To evaluate and analyze the risk factors for burning mouth syndrome (BMS). Eighty-seven consecutive patients with BMS and a randomly selected control group (n = 82) were comprehensively investigated with a self-designed questionnaire, Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS). A complete blood count and serum sex hormone were also examined in patients with BMS and control subjects. All the data obtained were transferred to a data bank and analyzed statistically in SPSS 11.5 for windows. No statistical difference between the BMS group and the control group was found in blood analyses including white blood cell count, red blood cell count, hemoglobin and platelet count. Among the menopausal or postmenopausal women with BMS, the follicle stimulating hormone (FSH) level was significantly higher, but the estradiol level was significantly lower. The BMS group reported adverse life events more frequently than the control group. Patients with BMS significantly exhibited symptoms of somatization, and both the scores of anxiety, depression in patients with BMS were higher than those of the control group (P < 0.05). A regression equation which included six variables had been established by using logistic regression analysis, indicating that the habit of tongue thrusting, lip sucking, periodontitis, smoking, outcome of recent medication, depression were the principal risk factors, among which tongue thrusting was the most significant. Our study indicated that BMS may be of psychological origin, and the measures such as refraining from oral parafunctional activities, removing local irritating factors, stopping smoking, good mental health status could help in the prevention of BMS.
  Journal of Oral Pathology and Medicine 01/2009; 38(1):24-8. · 1.63 Impact Factor
• Article: Replication and infectivity of hepatitis B virus in HBV-related glomerulonephritis.

  Lizhang Chen, Chengqiu Wu, Xuegong Fan, Jianming Gao, Hongling Yin, Tom Wang, Jun Wu, Shi Wu Wen
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  ABSTRACT: To examine the presence of hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), and hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) in renal tissues from patients with HBV-related glomerulonephritis. Renal tissue biopsies taken from patients with HBV-related glomerulonephritis and two control groups were prepared for immunocytochemical detection of HBsAg and HBcAg. HBV cccDNA was examined using a nested PCR. Of the 63 HBV-related glomerulonephritis patients studied, HBsAg was present in the renal tissues of 48 (76.2%) and HBcAg in the renal tissues of 27 (42.9%). The HBsAg and HBcAg positive rates in HBV-related glomerulonephritis patients were higher than those of the 20 patients with non-HBV-related glomerulonephritis (p<0.05). However, there was no significant difference when the HBV-related glomerulonephritis patients were compared with 12 patients with renal tuberculosis, renal atrophy, renal calculus, and renal tumor with positive serum HBV markers. In patients with HBV-related glomerulonephritis, there was no significant difference in HBsAg and HBcAg positive rates in renal tissue between patients with and without serum hepatitis B e antigen (HBeAg). By nested PCR, two of five patients with HBV-related glomerulonephritis were positive for HBV cccDNA. The location and replication of HBV in renal tissue make the kidney a potential reservoir for HBV. HBV cccDNA may be key in the search for anti-HBV drugs.
  International journal of infectious diseases: IJID: official publication of the International Society for Infectious Diseases 12/2008; 13(3):394-8. · 2.17 Impact Factor
• Article: Detection of hepatitis B surface antigen, hepatitis B core antigen, and hepatitis B virus DNA in parotid tissues.

  Lizhang Chen, Fuqiang Liu, Xuegong Fan, Jianming Gao, Nengzhi Chen, Tom Wong, Jun Wu, Shi Wu Wen
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  ABSTRACT: To examine the presence of hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), and hepatitis B virus (HBV) DNA in parotid tissues from patients with positive serum HBV markers. HBsAg and HBcAg were examined in parotid biopsy tissues from patients with suspected parotid tumor and positive serum HBV markers by immunocytochemistry, and HBV DNA was detected in parotid tissues by PCR. Among the 22 patients with a parotid tumor, only one was pathologically confirmed as a neoplasm; all others were benign. HBsAg and HBcAg were present in parotid cells with positive rates of 45.5% (10/22) and 40.9% (9/22), respectively, with an overall positive rate of 54.5% (12/22). Of the 22 cases with serum markers of HBV infection, seven (31.8%) had both HBsAg and HBcAg in the parotid cells. HBV DNA was present in seven of the 12 samples in which hepatitis B antigen was detected (58.3%). HBV in saliva might originate from the infected salivary glands and the infectious saliva could transmit HBV.
  International journal of infectious diseases: IJID: official publication of the International Society for Infectious Diseases 07/2008; 13(1):20-3. · 2.17 Impact Factor
• Article: [Gene modification in the genome of Epstein-Barr virus cloned as a bacterial artificial chromosome].

  Jianhong Lu, Yunlian Tang, Ming Zhou, Minghua Wu, Jue Ouyang, Jianming Gao, Liming Zhang, Dan Li, Qiong Chen, Wei Xiong, Xiaoling Li, Ke Tang, Guiyuan Li
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  ABSTRACT: Epstein-Barr virus (EBV) is an oncogenic herpesvirus associated with a variety of malignancies, including Burkitt's lymphoma and nasopharyngeal carcinoma (NPC). Functions of most EBV genes have not been determined. The use of bacterial artificial chromosome (BAC) to clone and modify the genome of EBV has enhanced the gene function study in the context of genome. Infectious clones of EBV were previously established by using EBV-BAC plasmid p2089. In order to further investigate EBV mutant biology, an easy and efficient method for gene modification in EBV-BAC was developed and detailed. The kanamycin gene (kan) flanked by recombinase FLP recognition targets (FRTs) was amplified from plasmid pKD13 and inserted into the vector of pcDNA3.1(+). Through the introduction of restriction endonuclease BsmB I in PCR primers, NPC-derived LMP1 gDNA containing the full-length ORF was then precisely ligated with kan on pcDNA3.1(+). The linear DNA segment of kan-LMP1 was transformed into E. coli DH10B cells containing p2089 and plasmid pKD46, homologous recombination was subsequently mediated by redalphabetagamma system from bacteriophage lambda. By this linear transformation and ET cloning, the full-length LMP1 in EBV-BAC (p2089) was replaced by the kan-LMP1. The introduced kan gene in EBV-BAC genome was eliminated specifically by the recombinase FLP when transformed by plasmid pCP20, leaving an FRT scar of 69 bp. The mutant could be identified by antibiotic screening and PCR amplification on bacteria medium. This method allows the gene of interest to be easily modified alone and then to be introduced into EBV-BAC genome. Following this example of gene substitution, other mutations such as deletion, insertion and point mutation become convenient work, and this improved method can be a potential use of gene modification in other BAC-based herpesvirus genome.
  ACTA MICROBIOLOGICA SINICA 04/2008; 48(3):385-90.
• Article: Epstein-Barr virus integrates frequently into chromosome 4q, 2q, 1q and 7q of Burkitt's lymphoma cell line (Raji).

  Jianming Gao, Xiaomin Luo, Ke Tang, Xiaoling Li, Guiyuan Li
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  ABSTRACT: Epstein-Barr virus (EBV) integration into a Burkitt's lymphoma (BL) cell line (Raji) was investigated, using polymerase chain reaction (PCR), Southern hybridization, genomic library screening and fluorescence in situ hybridization (FISH). BaMHIW fragments of the EBV genome and DNA sequences of the viral latent membrane protein (LMP)1 and LMP2 genes were detected in Raji cells. BaMHI-digested high-molecular weight DNA from Raji cells generated 4 and 10 kb, 23 kb fragments that hybridized to Probe-1 (EBV genome 13232-16189) and Probe-2 (EBV genome 5-3271). Genomic library for Raji cells was constructed. Plaques (1 x 10(5)) were screened with Probe-2, and four positive clones were obtained. Chromosomal integration of EBV DNA was detected in the Raji cell. The viral integration sites included 1p, 1q, 2q, 3p, 3q, 4q, 5q, 6q, 7p, 7q, 9q, 11p, 14q and 15q. Despite this multiplicity of integration sites, integration showed high frequency only at the sites 4q, 2q, 1q and 7q; 64% of the total signals were found in these four chromosomal bands. No viral integration occurred in chromosomes 16-22 or the sex chromosomes (X, Y). This study is the first comprehensive FISH analysis of EBV integration into the chromosomes of the Raji cell line. The findings support the notion that EBV integrates into the Raji cell genome non-randomly.
  Journal of Virological Methods 10/2006; 136(1-2):193-9. · 2.01 Impact Factor