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Dai D.-P.,Beijing Hospital and Beijing Institute of Geriatrics | Geng P.-W.,Wenzhou University | Geng P.-W.,The Laboratory of Clinical Pharmacy | Wang S.-H.,The Laboratory of Clinical Pharmacy | And 6 more authors.
Basic and Clinical Pharmacology and Toxicology | Year: 2015

Cytochrome P450 2D6 (CYP2D6) is one of the most widely investigated CYPs related to genetic polymorphisms and is responsible for one-quarter of the currently used clinical drugs. We previously detected 22 novel, non-synonymous, mutated sites in the Chinese population, but nothing is known about the functional effects of these mutations in terms of specific CYP2D6 substrates. In this study, wild-type CYP2D6, two common allelic variants and 22 newly reported CYP2D6 isoforms were transiently expressed in 293FT cells, and the enzymatic activities of these variants were systematically assessed using dextromethorphan and bufuralol as the probing substrates. Consequently, 19 and 21 allelic variants were found to exhibit significantly decreased enzymatic activities for dextromethorphan and bufuralol, respectively. Of 22 novel CYP2D6 variants, six allelic isoforms (CYP2D6.89, CYP2D6.92, CYP2D6.93, CYP2D6.96, E215K and R440C) exhibited absent or extremely reduced metabolic activities compared with those observed for the wild-type enzyme. Our in vitro functional data can be useful for CYP2D6 phenotype prediction and provide valuable information for the study of clinical impact of these newly found CYP2D6 variants in China. © 2015 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

Zheng M.Y.,The First Peoples Hospital of Wenling
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology | Year: 2010

To investigate the prevalence of transfusion-transmitted virus (TTV) and human papillomavirus (HPV) co-infection in cervical smears of patients with cervical lesions in littoral of Zhejiang province and analysis of transmitted route. Nested polymerase chain reaction (nPCR) was established. TTV DNA were tested by nPCR in cervical smears of 95 patients with cervical lesions and 55 healthy women, paired serum samples were available from 55 and 42 women, and their viral titer. The genotypes of 95 specimens of cervical cytology were detected with HybriMax. The phylogenetic group of TTV was determined by means of nPCR with N22 primers. The prevalence of TTV DNA in cervical smears of patients with cervical lesions and healthy women was 52.7% (29/55) and was comparable with that in paired serum sample (50%). Symptomatic women had significantly higher prevalence of TTV DNA in cervical smears (74.7%) than healthy controls (P = 0.005). The TTV DNA prevalence in patient serum samples was 51%. The phylogenetic groups of TTV serum isolates were concordant with those of TTV from cervical smears of the same subjects, and genotype was G1b. The TTV viral titer in cervical smears were 10 to 1000 times as high as in serum. The total infection rate of HPV was 98.9% in patients, and was 27.3% in healthy women. The frequently detected genotype was HPV16, 18, 33 of HSIL, and HPV6 of LSIL. The HPV positive study subjects had significantly higher TTV DNA prevalence than HPV negatives (P = 0.02). High prevalence of TTV in cervical smears suggests that sexual transmission is another mode of expansion of TTV infection among the population. The higher viral titer in cervical smears than in the respective serum samples might indicate active TTV replication in the female genital tract. Nevertheless, cooperation between TTV and HPV needs to be further investigated.

Wu Z.B.,The First Peoples Hospital of Wenling
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology | Year: 2010

To study the detection methods of BK virus infection in kidney transplant recipients, and to explore the clinical application. 132 cases of renal transplant recipients were undertaken BK virus detection including presence of decoy cells in urinary sediment, urine and serum BKV-DNA to demonstrate the BK virus replication. Among 132 cases of renal transplant recipients, urinary decoy cell was found in 37 (28.0%) patients and the median time was 12 months after surgery. 32 (24.2%) patients were diagnosed as BK viruria at a median of 11 months after surgery, and 16 (12.1%) recipients were diagnosed as BK viremia at a median of 15 months after surgery, 5 patients with BK viruria were diagnosed as BK virus associated nephropathy according to allograft biopsy. To make early diagnosis of BK virus infection, detection of urine decoy cells and BKV-DNA in urine and plasma sample is important,which provides an important basis for the prevention of BK virus associated nephropathy.

Yang B.S.,The First Peoples Hospital of Wenling
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology | Year: 2012

In order to understand the production mechanism of interferon and provide a scientific basis for preventionand clinical therapy, the expression changes of Toll-like receptor (TLR3) mRNA and the role of TLR3 in human lung epithelial cells (A549 cells) infected with respiratory syncytial virus (RSV) were investigated in this study. RSV infected A549 cells were treated with or without specific antibodies of TLR3 and collected at the selected timepoints after RSV infection (4, 8, 12, 16 and 24h). The expressions of TLR3, IFN-alpha, IFN-beta and RSV F mRNA were evaluated by RT-PCR. It was found that RSV infection could markedly up-regulate the mRNA expression of TLR3, IFN-alpha, IFN-beta and RSV F protein in a time-dependent manner as the 24h mRNA expressions of them were 4 times, 3 times, 3 times and 0.7 times more than the basic expression, respectively. Treatment of TLR3 specific antibodies, whereas, significantly down-regulated the activation of TLR3. The mRNA expression of IFN-alpha and IFN-1beta also decreased accordingly and that of IFN-beta reduced more obviously than IFN-alpha, but that of RSV F protein rose significantly. Above data indicate that RSV infection could induce an apparent increase of antiviral genes of IFN-alpha and IFN-beta by activating TLR3 in human lung epithelial cells and the activated cells mediated Type I interferon is antiviral, which suggesting that TLR3 might play an important role in antiviral activity of RSV-infected human lung epithelial cells.

Zheng M.Y.,The First Peoples Hospital of Wenling
Zhonghua fu chan ke za zhi | Year: 2010

To Investigate correlation between screening assay of human papillomavirus (HPV) and microbial pathogens in gynecology. Cervical samples were collected to search for HPV, bacteria and yeast infections in gynecologic outpatients. HPV typing was carried out by PCR and sequencing on cervical brush specimens. Chlamydia trachomatis was identified by strand displacement amplification (SDA) and the other microorganisms were detected by conventional methods. All data were analyzed to investigate the correlation among them. In this cross-sectional study, among 857 enrolled outpatients, there were 266 cases with positive HPV DNA, and the rate of infection was 31.0% (266/857). HPV genotype showed that thirty-five different HPV types were identified, of which HPV16 was the most prevalent (14.5%, 38/262), followed by HPV58 (9.2%, 24/262), HPV53 (8.0%, 21/262) and HPV42 (6.1%, 16/262); while other genotypes were present in less than 5% of HPV positive women. According to the reclassification, the aggregated percentage (high-risk and probably high-risk) of detected HPV was 58.8% (154/262), 27.9% (73/262) for low-risk and 13.4% (35/262) for unknown-risk HPV types. Among HPV positive women, cervical brush specimens results showed that more than 60% cases with normal cytology, 3.8% (10/266) with high-grade squamous intraepithelial lesions (HSIL), 29.7% (79/266) with low-grade squamous intraepithelial lesions (LSIL) and 3.0% (8/266) with atypical squamous cells of undetermined significance (ASCUS), respectively. Statistical analyses revealed there was a significant association between the infected HPV and Chlamydia trachomatis or Ureaplasma urealyticum (> 10,000 CCU/ml; all P < 0.01), while no correlation was found between HPV infection and bacterial vaginosis, streptococcus agalactiae, candida, Trichomonas vaginalis or Ureaplasma urealyticum (≤ 10 000 CCU/ml; all P > 0.05). Among the cases with bacterial vaginosis, the positive rate of HPV infected was 42.6%. Chlamydia trachomatis was one of the high-risk factors for the infection of HPV (OR = 2.82, 95% CI: 1.74 - 4.57). Mycoplasma hominis was isolated only in 2 cases, no patient was infected with Neisseria gonorrhoeae. Although bacterial vaginosis was not significantly associated with HPV, it was more common among the HPV positive women. There is the significant association between HPV and Chlamydia trachomatis or Ureaplasma urealyticum which may be increase the infection of HPV. These data suggest that it may be important to screen for the simultaneous presence of different microorganisms which may have synergistic pathological effects.

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