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Objective To explore the efficacy and safety of a raltegravir (RAL)-containing regimen among patients on methadone maintenance therapy.Methods From January 2010 to November 2010, 30 virus (HIV) treatment naive patients who were on methadone maintenance therapy were enrolled from a HIV clinic in Kunming, Yunnan Province and a HIV clinic in Hengyang, Hunan Province.All patients were given RAL, tenofovir (TDF) and lamivudine (3TC) as highly active antiretroviral therapy (HARRT).Patients were followed up for 48 weeks to evaluate the adjustment of methadone dose, opiate withdrawal reaction, antiretroviral efficacy and safety.Results From January 2010 to November 2010, 30 HIV patients were enrolled from the two appointed HIV clinics.The mean age was 39±6 years, with 73.3% male patients and 97% Han population.Before the treatment, their mean CD4+T lymphocyte counts was 210 /μL.Ninety percent of patients were co-infected with hepatitis C.Twenty-nine patients who completed study follow-up were included in final analysis.Five (17.8%) patients reported opiate withdrawal symptoms and increased methadone dose 4 weeks after HARRT.At 24 weeks and 48 weeks of HARRT, the average increase of CD4+T lymphocyte counts were (136±71) /μL and (185±88)/μL, respectively.Among patients who provided valid HIV-1 RNA testing results, 82.6% (19/23) and 95.8% (23/24) of patients had undetectable viral load at week 24 and week 48.Six grade 1-2 adverse events were reported in 4 patients.Conclusions In this pilot study, the new regimen containing RAL, TDF and 3TC appears to be an ideal option for patients on methadone maintenance therapy, because of its limited impact on methadone dose and good efficacy and safety profile.
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Objective To analyze the effect of long-term anti-viral treatment in children with acquired immune deficiency syndrome (AIDS) and investigate the factors affecting the treatment efficacy and growth and development of the children, so as to provide reference for improving the efficacy of antiviral drugs.Methods Children with AIDS receiving anti-retroviral treatment during 2004 to 2016 were retrospectively enrolled.The height, weight and CD4+ T cell counts were recorded every half year and the measurement of HIV RNA load was recorded on an annual basis.The CD4+ T cell counts and viral inhibition rates for the children who were under the treatment in the first year, 1~0.05);and viral inhibition rates were 92.9% and 97.6%, respectively with no statistical significance (χ2=1.071, P>0.05).The viral inhibition rate for the children receiving the treatment for 1 year was 85.7%, while that for whose treatment lasted for more than 10 years was 100.0%.A total of 5 cases developed drug-resistance (2 cases treated for 1 to 5 years and 3 cases for 5 to 10 years), and the virus replication was completely inhibited after switching to Lopinavir/ritonavir (LPV/r).The drug compliance was more than 95.0%.64.8% of children met the standard height, while 68.5% met the standard body mass.The baseline and last measured CD4+ T cell counts showed no significant differences between family-raised and social organization-raised children (Z=-1.159 and -0.523, respectively, both P>0.05).The children from high-income families had no significant differences compared with those from low-income ones in terms of the baseline and last measured CD4+ T cell counts (Z=-0.019 and -0.776, respectively, both P>0.05).Conclusions The long-term anti-retroviral treatment can effectively elevate the CD4+ T cell counts, inhibit viral replication and ensure drug compliance, which may promote the growth and development of children.However, approximately 30% children are still lower than the normal standards in terms of height and body mass.The drug-taking observer plays a central role on treatment effect.Most of the children′s family suffer from poor economic conditions.
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Aim To investigate the effect and mecha-nism of quercetin combined with cisplatin on prolifera-tion and apoptosis of human osteosarcoma cell line MG-63 . Methods MG-63 cells were treated with quercetin alone or combined with cisplatin. Cellular morphologic changes were observed under inverted phase contrast microscope. The effects of proliferation inhibition were assayed by CCK-8 method. The combination effect was judged through Chou-Talaly analysis. The apoptosis ra-tios of cells were analyzed by flow cytometry. The gene expression of Bcl-2 and caspase-3 was detected by RT-PCR assay. The protein expression of Bcl-2 and caspase-3 was measured by Western blot assay. Re-sults Quercetin alone or combined with cisplatin could inhibit the proliferation, but induce the apoptosis of MG-63 cells. Combination of quercetin and cisplatin revealed a synergistic effect on cell proliferation and apoptosis as it reduced the expression of Bcl-2 but en-hanced that of caspase-3 at both gene and protein lev-els. Conclusion Synergistic effect of quercetin com-bined with cisplatin on cell proliferation and apoptosis of MG-63 cells is possibly due to reduction of Bcl-2 and enhancement of caspase-3 expression.
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The gene encoding the VP28 envelope protein of White spot syndrome virus (WSSV)was cloned into expression vector pET-30a and transformed into the Escherichia coli strain BL21.After induction,the recombinant VP28 (rVP28) protein was purified and then used to immunize Balb/c mice for monoclonal antibody (MAb) production.It was observed by immuno-electron microscopy the MAbs specific to rVP28 could recognize native VP28 target epitopes of WSSV and dot-blot analysis was used to detect natural WSSV infection.Competitive PCR showed that the viral level was approximately 104 copies/mg tissue in the dilution of gill homogenate of WSSV-infected crayfish at the detection limit of dot-blot assay.Our results suggest that dot-blot analysis with anti-rVP28 MAb could rapidly and sensitively detect WSSV at the early stages of WSSV infection.
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The M and S segment cDNAs of hantavirus Z5 strain was amplified by RT-PCR,and the purified PCR products were cloned into vector pGEM-T and then sequenced.It was demonstrated that the M genome segment of Z5 was found to be 3 616 nucleotides in length with a single open reading frame encoding 1 135 amino acids.And the S genome segment was 1700 nucleotides in length with a single open reading frame encoding 429 amino acids.As demonstrated by the homologous analysis of nucleotides and amino acids,it was showed that the Z5 strain belonged to hantaan viruses HTN type and was the same subtype of the Z10 strain.It is conclouded that difference in nucleotide sequence exists between Z5 strain with other Hantavirus strains but high level of homology in amino acid sequences is still present.
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One strain of Hantavirus(HV) was isolated from patients with hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province and its S-segment was cloned and submitted to nucleotide sequence analysis in order to determine the type of strain and extent of genetic variation for further study on its evolution and mutation .The HV antigen was detected from mouse lungs in endemic area by direct immunofluorescene test and the HV-positive sample of mouse lung was inoculated to Vero-E6 cells to isolate virus. The total cellular RNA was extracted from infected cell culture and the S segment gene was amplicated by RT-PCR. Then, the purified PCR product was cloned into T vector for sequencing. The result showed that the full-length sequence of the S segment was 1 700 bp with one open reading frame (ORF) encoding 429 amino acids. Comparison with Hantavirus HTN type showed 85.7%-91.9% homology at the nucleotide level. In comparison with the SEO type of viruses, the homology at nucleotide level was shown to be 71.2%-75%. This new HV strain may be typed as to HTN virus and may be a new subtype of this virus.
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Objective To study the complete genome sequence of Hantavirus ZT71 strain gene isolated in Zhejiang province and explore its evolution. nethods The total RNA was prepared from ZT71 virus infected cells and the RT-PCR products were cloned into T vector, sequenced and analyzed. Results The L, M and S segments of the strain ZT71 genome were 6530,3651 and 1753 nucleotides in length with a single open reading frame individually encoding 2151,1133 and 429 amino acids. The sequence analysis of nucleotides showed that the homology of L, M and S segments of strain ZT71 between those of other strains of Seoul virus could reach 95.5%-99.7%, 84.1%-99.6% and 88.7%-99.5%, respectively. The analysis of the deduced amino acids showed the similar result. The source of strain ZT71 could be traced from the analysis of the phylogentic trees of nucleotides and amino acids, and it should belong to Seoul type of Hantavirus which was also verified serologically. Conclusion The nucleotide sequences and deduced amino acid sequences of L, M and S segments of strain ZT71 are similar to that of those of Seoul type of Hantavirus. And Hantaan type virus used to be prevalent primarily in Zhejiang province,and it would be an endemic area of mixed type of Hantavirus since the discoveries of the viruses of Soeul type in recent years.