[Prevalence status and mutation pattern of H221Y in subtype B' of HIV-1].

OBJECTIVE: To explore the prevalence and mutation pattern of H221Y at reverse transcriptase (RT) among the subtype B' of human immunodeficiency virus1 (HIV-1) in antiviral therapy-failure patients. METHODS: A total of 1363 sequences, comprising of 1205 therapy-failure individuals and 158 therapy-naive individuals, were submitted to the Stanford HIV drug resistance database (SHDB) to analyze the frequency and mutation pattern of H221Y. RESULTS: The prevalence of mutation H221Y in the therapy-failure population was significantly higher than that of the therapy-naive (6.59% vs 0.60%) (χ(2) = 6.59, P = 0.027). The emergence of H221Y usually accompanied the position mutations of T215, M184, K103 and Y181 of RT, and the pattern of TAMs/H221Y/Y181C/I was common. Frequency of H221Y in the regimen of AZT/ddI/NVP was more popular than the other 4 regimens (14.6% vs 3.5%, 4.9%, 2.3%, 2.6%, all P < 0.01). CONCLUSION: With a unique mutation pattern, H221Y has a low prevalence in the individuals of first-line therapy-failure patients.

[Occurrence of human immunodeficiency virus-1 resistance through a six-year surveillance in rural areas of Henan].

OBJECTIVE: To analyze the occurring rules of human immunodeficiency virus (HIV) drug resistance under an unique therapy model among HIV-1 infected individuals on antiretroviral therapy (ART) in rural areas of Henan, China. METHODS: A cohort of 75 individuals on an ART regimen of zidovudine (ZDV), dideoxyinosine (ddI) and nevirapine (NVP) was established in March 2003. A total of 12 surveillance were conducted and 788 person-times were studied until 2010. The parameters of CD4 cell count and viral load (VL) were tested in each survey. And genotypic resistance testing was performed in patients with a failure of viral suppression. Survival analysis was used to estimate the occurrence time of resistance. RESULTS: The cumulative mortality rate was 16% (12/75) in the cohort. And the cumulative resistance rate was 88% (66/75) from 2004 to 2010. The rate of resistance reached 54.7% and the probability from susceptibility to drugs developing resistance decreased drastically from 100% to 45.3% within the first 1 year of initiation. The occurrence time of resistance for half of individuals in the cohort was at 12.0 months (95%CI 8.6 - 17.0) after initiation, 25.1 months (95%CI 19.0 - 33.3) in those whose VL was less than 4.0 lgU/ml and 4.8 months (95%CI 4.1 - 5.6) at VL > 4.0 lgU/ml during the first investigation. The individuals with an early occurrence of resistance within 12 months carried high risks for a failure of viral suppression and a decrease of CD4 counts. CONCLUSION: The occurrence of resistance rises with the course of therapy. And the greatest probability for resistance is within the first 1 year of initial therapy. A high level of VL has a significant impact on the development of resistance. Preventing the occurrence of resistance during the initial therapy remains a key goal.

UGT1A1 rs4148323 A Allele is Associated With Increased 2-Hydroxy Atorvastatin Formation and Higher Death Risk in Chinese Patients With Coronary Artery Disease.

It is widely accepted that genetic polymorphisms impact atorvastatin (ATV) metabolism, clinical efficacy, and adverse events. The objectives of this study were to identify novel genetic variants influencing ATV metabolism and outcomes in Chinese patients with coronary artery disease (CAD). A total of 1079 CAD patients were enrolled and followed for 5 years. DNA from the blood and human liver tissue samples were genotyped using either Global Screening Array-24 v1.0 BeadChip or HumanOmniZhongHua-8 BeadChip. Concentrations of ATV and its metabolites in plasma and liver samples were determined using a verified ultra-performance liquid chromatography mass spectrometry (UPLC-MS/MS) method. The patients carrying A allele for the rs4148323 polymorphism (UGT1A1) showed an increase in 2-hydroxy ATV/ATV ratio (p = 1.69E-07, false discovery rate [FDR] = 8.66E-03) relative to the value in individuals without the variant allele. The result was further validated by an independent cohort comprising an additional 222 CAD patients (p = 1.08E-07). Moreover, the rs4148323 A allele was associated with an increased risk of death (hazard ratio [HR] 1.774; 95% confidence interval [CI], 1.031-3.052; p = 0.0198). In conclusion, our results suggested that the UGT1A1 rs4148323 A allele was associated with increased 2-hydroxy ATV formation and was a significant death risk factor in Chinese patients with CAD.

[Clinical analysis of 48 patients with vaginal enlarged amputation of cervix].

OBJECTIVE: To evaluate the feasibility and safety of vaginal enlarged amputation of cervix to treat patients with cervical cancer of stage Ia1 and cervical intraepithelial neoplasia grade III (CIN III) who were unfit for conization surgery. METHODS: From July 2002 to May 2007, patients with cervical cancer at stage Ia1, diagnosed by pathology after loop electrosurgical excision procedure (LEEP), large area CIN III (the area of lesion>or=3/4 on colposcopy), CIN III coexisted with vaginal intraepithelial neoplasia (VAIN) in the superior segment of vagina, CIN II-III recurrence or with residual lesion, positive margin after conization of cervix, who wanted to preserve fertility and (or) corpus uteri were selected to receive vaginal enlarged amputation of cervix. RESULTS: Forty-eight cases including 5 with cervical cancer in stage Ia1, 38 with large area CIN III (9 with gland involvement), 2 with residual lesion and 2 with positive margin after LEEP, 1 recurrence after cold knife conization, received the procedure successfully. The median age was 34 years (range 27-40), median operation time was 60 minutes (range 30-100), median blood loss was 40 ml (range 5-300), and median hospital stay was 10 days (range 7-17). After follow-up 1-39 months, no patient had postoperative complications and recurrence, and all patients resumed normal menstrual cycle and sexual life. CONCLUSION: Vaginal enlarged amputation of cervix appears to be a safe and feasible procedure for patients with cervical cancer at stage Ia1 and CIN III who are unfit for conization surgery.

Phenotypic resistance of resistant strains of HIV type-1 subtype B in China.

BACKGROUND: This study was aim to explore the characteristics of phenotypic resistance of resistant strains of HIV type-1 (HIV-1) subtype B and to compare the concordance between the phenotypic resistance and genotypic resistance. METHODS: The genotypic resistance assay for the HIV-1 clinical isolates was performed. One isolate without resistance mutation was chosen as a drug-sensitive reference strain and seven subtype B isolates with resistance mutations were phenotypically tested. Fifty percent inhibitory concentrations (IC50) between resistant and sensitive viruses were compared. The resistance extent was determined by the folds of the increased IC50. The concordance between the phenotypic resistance and genotypic resistance was also analyzed. RESULTS: IC50 of resistant isolates were 0.0006 - 0.1300 micromol/L for zidovudine (AZT), 0.0016 - 0.0390 micromol/L for lamivudine (3TC), 0.0104 - 0.4234 micromol/L for nevirapine (NVP), and 0.0163 - 0.1142 micromol/L for indinavir (IDV), respectively. Genotypic and phenotypic resistance assays indicated that the resistant strains were intermediately and highly resistant to nucleotide analog reverse transcriptase inhibitors and non-nucleotide analog reverse transcriptase inhibitors. The phenotypic assay was consistent with the genotypic assay. For measuring the potential resistance, the genotypic assay was more sensitive than the phenotypic. In evaluating the resistance to protease inhibitors, these two assays were discrepant. CONCLUSIONS: Both the phenotypic and genotypic assays indicate that the resistant viruses exist in HIV-infected patients in China who have received treatment. Phenotypic and genotypic assays have high concordance, and the genotypic assay could replace the phenotypic assay to predict the HIV-1 resistance.

Plasma miR-142 accounting for the missing heritability of CYP3A4/5 functionality is associated with pharmacokinetics of clopidogrel.

AIM: To investigate whether plasma miRNAs targeting CYP3A4/5 have an impact on the variance of pharmacokinetics of clopidogrel. MATERIALS & METHODS: The contribution of 13 miRNAs to the CYP3A4/5 gene expression and activity was investigated in 55 liver tissues. The association between plasma miRNAs targeting CYP3A4/5 mRNA and clopidogrel pharmacokinetics was analyzed in 31 patients with coronary heart disease who received 300 mg loading dose of clopidogrel. RESULTS: Among 13 miRNAs, miR-142 was accounting for 12.2% (p = 0.002) CYP3A4 mRNA variance and 9.4% (p = 0.005) CYP3A5 mRNA variance, respectively. Plasma miR-142 was negatively associated with H4 Cmax (r = -0.5269; p = 0.0040) and associated with H4 AUC0-4h (r = -0.4986; p = 0.0069) after 300 mg loading dose of clopidogrel in coronary heart disease patients. CONCLUSION: miR-142 could account for a part of missing heritability of CYP3A4/5 functionality related to clopidogrel activation.

Prevalence and evolution of drug resistance HIV-1 variants in Henan, China.

To understand the prevalence and evolution of drug resistant HIV strains in Henan China after the implementation of free antiretroviral therapy for AIDS patients. 45 drug naïve AIDS patients, 118 AIDS patients who received three months antiretroviral therapy and 124 AIDS patients who received six months antiretroviral treatment were recruited in the southern part of Henan province. Information on general condition, antiretroviral medicines, adherence and clinical syndromes were collected by face to face interview. Meanwhile, 14 ml EDTA anticoagulant blood was drawn. CD4/CD8 T cell count, viral load and genotypic drug resistance were tested. The rates of clinical improvement were 55.1% and 50.8% respectively three months and six months after antiretroviral therapy. The mean CD4 cell count after antiretroviral therapy was significantly higher than in drug naïve patients. The prevalence rate of drug resistant HIV strains were 13.9%, 45.4% and 62.7% in drug naïve patients, three month treatment patients and six month treatment patients, respectively. The number of resistance mutation codons and the frequency of mutations increased significantly with continued antiretroviral therapy. The mutation sites were primarily at the 103, 106 and 215 codons in the three-month treatment group and they increased to 15 codon mutations in the six-month treatment group. From this result, the evolution of drug resistant strains was inferred to begin with the high level NNRTI resistant strain, and then develop low level resistant strains to NRTIs. The HIV strains with high level resistance to NVP and low level resistance to AZT and DDI were highly prevalent because of the AZT+DDI+NVP combination therapy. These HIV strains were also cross resistant to DLV, EFV, DDC and D4T. Poor adherence to therapy was believed to be the main reason for the emergence and prevalence of drug resistant HIV strains. The prevalence of drug resistant HIV strains was increased with the continuation of antiretroviral therapy in the southern part of Henan province. Measures, that could promote high level adherence, provide new drugs and change ART regimens in failing patients, should be implemented as soon as possible.

[A cross-sectional study on the prevalence of HIV drug resistance in patients receiving antiretroviral treatment in Shenqiu county, Henan province].

OBJECTIVE: To understand the prevalence of drug resistance in AIDS patients who had been receiving HAART in a long run, in Shenqiu county, Henan province. METHODS: This cross-sectional study included 120 HIV infected patients who began receiving ART (antiretroviral therapy) in 2003. Viral loads and CD(4)(+) T cells counts were measured, and In-house drug resistance test was performed in VL > 1000 copies/ml patients. RESULTS: 114 cases out of 120 patients had complete viral load data. Among them, 33 cases having viral loads less than 50 copies/ml, and the remaining viral loads showed an average of lg (4.09 ± 1.10) copies/ml. The average of CD(4)(+) T cell counts was (377 ± 218) cells/ml, with 64 (53.3%) cases showing their CD(4)(+) T cell counts higher than 350 cells/ml. In 67 patients, 58 of them showed genotypic resistance, and 40 cases showed reverse transcriptase inhibitors (RTIs) resistance. The ratios of nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) resistance were 53.4% (31/58) and 67.2% (39/58), respectively. There were no differences of drug resistance ratio in the three treatment programs. The highest drug resistance rates in NRTIs and NNRTIs were zidovudine, lamivudine, nevirapine. However, protease inhibitors (PIs) resistance variants were not found. CONCLUSION: The prevalence of drug-resistant strains seemed to be high in Shenqiu country, Henan province. Long-term follow-up monitoring strategy should be developed to optimize the timely treatment programs.

Similar neutralizing activity in the HIV-1 infected long term non-progressors(LTNPs) and typical progressors(TPs).

Neutralizing antibodies are considered to be an important protective parameter used in HIV-1 vaccine evaluation. However, the exact role that neutralizing antibodies plays in controlling the disease progression of HIV-1 infected peoples is still undetermined. In this paper, we compared the protective function of the neutralizing antibody response in the plasma from LTNP and TP against clade B and clade C pseudoviruses. No difference in the neutralizing activities between the plasma from LTNP and TP was found, which was consistent with the most recent reports. In addition, no correlations between the titer or breadth and CD(4+) or viral load in HIV-1 infected individuals were found. The protective roles played by neutralizing antibodies in controlling disease progression of HIV-1 infected people need to be considered in a new viewpoint.

[Screening program on novel drug resistance mutations of subtype B' in human immunodeficiency virus type 1 in China].

OBJECTIVE: To screen the level of novel drug resistance mutations in subtype B' in China. METHODS: 451 pol sequences collected from the previous study, which including 354 AIDS patients who had received antiretroviral treatment (ART) and 97 the untreated patients. Entire protease gene (codons 1 - 99) and full-length reverse transcriptase gene (codons 1 - 560) were included. Variation of mutations between the treated and the untreated patients with consensus/ancestral sequences were compared and the mutations with higher frequencies in the treated patients than in the untreated patients were screened before submitting the mutations to the Stanford HIV Drug Resistance Database (SHDB) (http: //hivdb.stanford.edu/). Relation between the mutations and resistance preliminarily was then analyzed, according to the information including SHDB. RESULTS: Frequencies of 7 mutations at 6 positions, D123E, V292I, K366R, T369A, T369V, A371V and I375V, 2 at DNA polymerase domain and 5 at connection domain of reverse transcriptase (RT) were higher in the treated patients than in the untreated patients. The information of 7 mutations including the SHDB showed that 7 mutations were major variants at corresponding positions, and theirs frequencies were higher in the treated patients using some drugs, than in the untreated patients. CONCLUSION: 7 mutations being screened from the China subtype B were possibly associated with the resistance, which called for the construction of mutated viruses by site-directed mutagenesis to identify their effects on the susceptivity of different drugs.

[Studying on the prevalence and mutation pattern of N348I which related to the resistance of HIV-1].

OBJECTIVE: To elucidate the prevalence and the mutation pattern of N348I that related to the resistance seen in the AIDS patients, in China. METHODS: Partial pol gene of HIV-1 comprising of full protease (PR) and reverse transcriptase (RT) was obtained from plasma samples of therapy-failure individuals (n = 614) and therapy-naive individuals (n = 619) by using reverse transcription polymerase chain reaction (RT-PCR). 1233 sequences were then submitted to the HIV-1 drug resistance database of the Stanford University to analyze the prevalence and the emergence pattern of N348I. RESULTS: The prevalence of N348I was 6.5% in the therapy-failure patients and 0.8% in the naive individuals, respectively. The prevalence of N348I was more popular among those patients whose ART regimens containing zidovudine (AZT or ZDV) than those without AZT in regimens (14.1% vs. 4.7%, χ² = 10.21, P < 0.01). N348I always emerged, and combined with others mutations among patients of ART, whose frequencies were: 85.0% in combination with thymidine analog mutations (TAMs) and 52.5% with M184V/I, respectively. CONCLUSION: N348I was somehow prevalent in the therapy-failure patients when using the first-line antiretroviral drugs, and it emerged as unique patterns. This study laid the ground in improving the technology of drug resistance genotypes detection and providing theoretical basis to study the mechanism of resistance and the law of molecular evolution.

Indinavir resistance evolution in one human immunodeficiency virus type 1 infected patient revealed by single-genome amplification.

UNLABELLED: Human Immunodeficiency Virus Type 1 exists in vivo as quasispecies, and one of the genome's characteristics is its diversity. During the antiretroviral therapy, drug resistance is the main obstacle to effective viral prevention. Understanding the molecular evolution process is fundamental to analyze the mechanism of drug resistance and develop a strategy to minimize resistance. OBJECTIVE: The molecular evolution of drug resistance of one patient who had received reverse transcriptase inhibitors for a long time and had treatment which replaced Nevirapine with Indinavir was analyzed, with the aim of observing the drug resistance evolution pathway. METHODS: The patient, XLF, was followed-up for six successive times. The viral populations were amplified and sequenced by single-genome amplification. All the sequences were submitted to the Stanford HIV Drug Resistance Database for the analysis of genotypic drug resistance. RESULTS: 149 entire protease and 171 entire reverse transcriptase sequences were obtained from these samples, and all sequences were identified as subtype B. Before the patient received Indinavir, the viral population only had some polymorphisms in the protease sequences. After the patient began Indinavir treatment, the variants carrying polymorphisms declined while variants carrying the secondary mutation G73S gained the advantage. As therapy was prolonged, G73S was combined with M46I/L90M to form a resistance pattern M46I/G73S/L90M, which then became the dominant population. 97.9% of variants had the M46I/G73S/L90M pattern at XLF6. During the emergence of protease inhibitors resistance, reverse transcriptase inhibitors resistance maintained high levels. CONCLUSION: Indinavir-resistance evolution was observed by single-genome amplification. During the course of changing the regimen to incorporate Indinavir, the G73S mutation occurred and was combined with M46I/L90M.

Allele-specific real-time PCR testing for minor HIV-1 drug resistance mutations: assay preparation and application to reveal dynamic of mutations in vivo.

BACKGROUND: It is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20% - 30%. The aim of this study was to evaluate allele-specific real-time PCR (ASPCR) to detect the resistance-related mutations located at positions 103, 184 and 215. METHODS: We developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140 clinical samples using this method. RESULTS: The sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M184I, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained. CONCLUSIONS: Drug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Y emerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.

Competitive capacity of HIV-1 strains carrying M184I or Y181I drug-resistant mutations.

BACKGROUND: Virus with nucleoside reverse transcriptase inhibitors (NRTIs) or nonnucleoside reverse transcriptase inhibitors (NNRTIs) resistant mutations show different evolution tendencies when the anti-viral therapies are interrupted. Understanding the replication fitness of drug-resistant virus is important for the study of the prevalence of drug-resistance. For this purpose, we characterized the replication capacity of HIV-1 virus carrying lamivudine (3TC) or nevirapine (NVP) resistant mutations. METHODS: 3TC and NVP resistant variants were induced in vitro by selecting wild type virus in the presence of drugs. For the competitive replication assay, drug-resistant variants were cocultured with wild-type virus in the presence or absence of drugs. The ratios of the viral species were determined over time by using a real-time RT-PCR-based assay. RESULTS: 3TC-resistant (M184I mutation) and NVP-resistant (Y181I mutation) virus should be selected in vitro in two different ways. The competitive replication assay showed that the ratio of virus carrying a M184I mutation increased from 98.8%, while the wild type virus decreased to 1.2% after 4 passages in the presence of 3TC; the percentage of virus carrying the Y181I mutation increased to 90.5%, while wild type virus decreased to 9.5% in the presence of NVP. In the absence of drugs, the ratio of virus carrying the M184I mutation decreased to 5.3%, while wild type virus increased to 94.7%; the ratio of virus carrying Y181I increased to 75%, while wild type virus decreased to 25% after 4 passages. CONCLUSIONS: The NVP-resistant virus is fitter than wild type virus even in the absence of NVP that may be the reason that NNRTIs-resistant virus is spreading quickly.

[Concurrent chemoradiotherapy versus radiotherapy in advanced cervical carcinoma].

BACKGROUND & OBJECTIVE: Cisplatin-based concurrent chemoradiotherapy has become the standard treatment modality for locally advanced cervical cancer. However, the optimal chemotherapy regimen combined with radiotherapy remains controversial. This study was to compare the therapeutic efficacy and toxicity of concurrent chemoradiotherapy with those of radiotherapy, and those among different regimens of concurrent chemoradiotherapy for stage IIB-IIIB cervical cancer. METHODS: From Jan. 2003 to Dec. 2004, 285 patients with stage IIB-IIIB cervical cancer treated in Maternal and Child Health Hospital of Jiangxi Province were randomly assigned to receive radiotherapy alone or concurrent chemoradiotherapy. According to different chemotherapy regimens, patients in the concurrent chemoradiotheapy group were randomly chosen to receive radiotherapy with chemotherapy of bleomycin and cisplatin (RT+BP), radiotherapy with chemotherapy of taxol and carboplatin (RT+TP), and radiotherapy with chemotherapy of 5-fluorouracil and cisplatin (RT+FP). The 3-year survival rates and toxicity of different groups were compared. RESULTS: After a median follow-up of 42 months, the 3-year survival was higher in the concurrent chemoradiotheray group (75%) than in the radiotherapy group (65%) (P=0.042). Acute treatment-related toxicity (grade III and IV) was higher in the concurrent chemoradiotherapy group than in the radiotherapy group (P<0.001); while the delayed treatment-related toxicity was similar in the two groups (P=0.613). The 3-year survival rates of BP, TP and FP chemoradiotherapy groups were 74%, 80% and 71%, without significant differences (P=0.792). Acute toxicities (grade III and IV) and delayed toxicities were similar among the three groups. CONCLUSIONS: Concurrent chemoradiotherapy significantly improves the survival for patients with stage IIB-IIIB cervical cancer compared to radiotherapy alone. Among the three chemoradiotherapy regimens, radiotherapy combined with taxol and carboplatin exerts a slightly higher 3-year survival than the other two regimens with tolerable toxicity.

[Study on the difference of the APOBEC3G mRNA levels among HIV long-term nonprogressors, slow progressors, people at high risks and normal people].

OBJECTIVE: To examine the APOBEC3G (hA3G) mRNA levels of four different groups in the human immunodeficiency virus (HIV) prevalent areas in central China and to analyze the relationship between hA3G mRNA levels and HIV disease progression. METHODS: We collected peripheral blood and isolated the peripheral boold monouuclear cells (PBMCs), and then cryo-preserved the PBMCs in liquid nitrogen. Prior to the total extraction of RNA, PBMCs were resuscitated and mRNA were reverse Transcripted to cDNA in vitro. Real-time polymerase chain reaction (PCR) was used to test hA3G mRNA levels of different groups. RESULTS: There were 13 HIV long term non-progressors with the mean CD4+ T lymphocyte count as (716 +/- 169) per microl and the mean affection time as (12.5 +/- 2.3) years. There were 48 HIV slow progressors with the mean CD4+ T lymphocyte count as (233 +/- 144) per microl and the mean affection time as (10.7 +/- 2.2) years. The hA3G mRNA level of HIV long term nonprogressors was higher than that of normal people while the hA3G mRNA level of HIV slow progressors was higher than that of normal people and high risk people. There were no correlations between CD4+ T lymphocyte count and hA3G mRNA levels of HIV long-term nonprogressors as well as in HIV slow progressors. CONCLUSION: There was difference found in the hA3G mRNA levels of four groups in the HIV popular area in central China while no correlation between CD4+ T lymphocyte count and hA3G mRNA levels of HIV long term nonprogressors as well as in HIV slow progressors were found.

[Research on the selective kinetics of HIV-1 nucleoside reverse transcriptase inhibitor drug resistance-associated mutations among 4 AIDS patients receiving highly active antiretroviral therapy].

OBJECTIVE: To elucidate the molecular evolutional characteristics of HIV-1 nucleoside reverse transcriptase inhibitor (NRTI) drug resistance-associated mutations in patients with AIDS receiving highly active antiretroviral therapy. METHODS: We selected 4 AIDS patients receiving highly active antiretroviral therapy (HAART) with good adherence under a HIV-1 drug resistance cohort from a rural region in central China. Those people carried susceptible virus at the beginning of treatment and gradually came to produce virus resistant to NRTIs during the process of antiretroviral therapy (ART). Reverse transcriptase (RT) genes from each patient's peripheral blood samples (from 3 to 33 months after withdrawal) were cloned and sequenced in succession. RESULTS: We sequenced a total number of 855 clones and obtained the HIV-1 NRTI drug resistance-associated mutations patterns of the 4 patients. Typical resistance mutations of thymidine analogue mutations (TAMs) pattern 1, such as L210W, T215Y and M41L, were generated in patient 'A'. TAMs pattern 2, including D67N, K70R and K219Q mutations, was discovered in patient 'B'. Interestingly, in patient 'C', some clones comprising not only TAMs pattern 1 mutations (T215Y) but also TAMs pattern 2 mutations (K70R, D67N). CONCLUSION: The four patients show different pathways on HIV-1 NRTI drug resistance-associated mutations, including TAMs pattern 1, TAMs pattern 2 and the fusion pattern of TAMs-1 & TAMs-2. We also noticed that the tendency of gradual accumulation was obvious and those mutations detected earlier tended to be the predominant strains.

[Study on the effect of treatment and the status of drug resistance among HIV/AIDS patients treated with highly active antiretroviral therapy in Shandong province].

OBJECTIVE: To understand the effect of highly active antiretroviral therapy (HAART) on AIDS patients, and to explore the prevalence and the impact of HIV-1 drug resistance in Shandong province. METHODS: 2 cross-sectional studies were carried on in 2004 and 2005, to collect data on clinical symptoms and compliance of the AIDS patients with HAART through questionnaire. Informed-consent principle was followed to test on immunological, viral and laboratory index of them. HIV-1 drug genotype resistance by sequencing the gene of HIV-POL after RT-PCR was performed and analyzed. RESULTS: 31 AIDS cases with and. 27 AIDS cases without HAART, were studied. 83.3% and 64.5% of the AIDS patients with HAART showed that the CD4+ T cell count was rising to over 350/microl, in the first study (2004) and in the second (2005) study respectively but still 45.8% and 45.2% of AIDS patients under HAART in the 2 years showed a decreasing HIV load under the detected limit. However, these findings were showing remarkable difference when compared with the AIDS without HAART. 7 drug resistance gene sites were found in AIDS patients with HAART and in AIDS patients without HAART. The rate on high degree drug resistance mutation and total drug resistance rate of mutation of the former were higher remarkably than those of the latter. CONCLUSION: Most of the AIDS patients with HAART met the purpose of rebuilding immunity and control of HIV,as well as alleviation of symptoms. Although the drug resistance stain appeared in Shandong,but had little effect on HAART. AIDS; Drug resistance; Highly active antiretroviral therapy

[Study on the antiviral therapy program among people with human immunodeficiency virus in Guangxi Zhuang Authority].

OBJECTIVE: To explore the reconstitution of immune function and viral suppression condition and to analyze the occurrence of drug resistance HIV-1 variants and its prevalence after using HAART in Guangxi Autonomy Region. METHODS: From July 2004 to October 2005, 133 HIV infected individuals who had received HAART for more than three months were recruited, and 58 infected persons with no antiviral therapy were selected as controls. Questionnaire was used to collect information about the adherence of HAART therapy. Immune reconstruction and viral suppress conditions were obtained by CD4+ counts and viral load and RT-PCR were used to amplify the PR and RT regions of HIV-1 genome while HIV-1 drug resistance rates were analyzed to show the occurrence and prevalence in both treated and naive patients. RESULTS: In terms of CD4+ T cell counts: 70.69% of the treated patients showed obvious increase and 23.28% had no apparent change but 6.03% of them went down. 70.48% of the patients who had received antiviral therapy more than 3 months had their viral load lower than the low detectable limitation. When comparing the log of viral load between treated and untreated cohort, the mean value of the treated was obviously less than the untreated (P < 0.05). However,the result of drug resistance showed no obvious difference between the treated and untreated groups. CONCLUSION: The antiviral therapy being used in Guangxi region, had achieved obvious effect on the reconstruction of immune system and the suppression of viral replication in vivo under good adherence while the occurrence of drug-resistant HIV strain did not show obvious difference between treated and naive patient groups.

An immunogenicity study of a newly fusion protein Cna-FnBP vaccinated against Staphylococcus aureus infections in a mice model.

Adhesins are considered the most important virulence factors during early phase Staphylococcus aureus infection. The present report describes a newly fusion protein, named Cna-FnBP that was constructed by fusion of the fnb and cna genes of S. aureus and expressed in E. coli. The recombinant protein was designed to broaden the function spectrum of block binding activity to S. aureus adhesion. Vaccination of the recombinant protein induced a strong and specific humoral response to Cna-FnBP in mice. In addition, splenocyte proliferation was provoked by in vitro stimulation with recombinant Cna-FnBP, thus, indicating direct implication of these cells in the immune response. These pre-incubated bacteria were phagocytosed by neutrophils at an increased level in vitro in a mouse model. Mice immunized with Cna-FnBP survived significantly longer following the challenge with S. aureus than nonimmunized mice did. These results indicate that Cna-FnBP is a promising vaccine for the prevention of S. aureus infections. Overall, the results suggest that fusion compounds which elicted from ECM-binding proteins (ECMBPs) were used to immunize against adhesins represents a valuable approach to combat S. aureus infections.