Objective: To analyze the variation characteristics of HIV-1 Gp120 sequences in men who have sex with men (MSM) in Guangzhou. Methods: Plasma samples were collected from HIV-1 infected MSM before antiretroviral treatment. Viral RNA was extracted from plasma. Gp120 gene sequences were amplified by reverse transcription and nested-PCR using specific primers. Phylogenetic tree, length polymorphism, amino acid characteristics of V3 loop, co-receptors and signature amino acids were analyzed. Results: The phylogenetic tree were divided into 4 clusters, and the most prevalent subtypes were CRF07_BC (34/61, 55.74%) and CRF01_AE (24/61, 39.34%). Majority of HIV-1 Gp120 sequences had 496-515 amino acids. Among five hypervariable regions, the V1 region had the highest levels of length polymorphism and V3 region had the lowest. The top four peptide of V3 loop were GPGQ (56/58, 96.55%). Most of the co-receptors HIV-1 strains used was CCR5(50/58, 86.21%)according to four methods of comprehensive prediction. There are four signature amino acids in CRF01_AE subtype strains, and the frequency of occurrence was 0.75-0.83; there are eight signature amino acids in CRF07_BC subtype strains, and the frequency was 0.74-0.94. Conclusions The length of Gp120 sequences in MSM in Guangzhou has a high polymorphism. The top four peptide of V3 loop, co-receptor and signature amino acid of V3 ring have formed unique patterns.

Objective To investigate the prevalence and characteristics of non-nucleoside reverse transcriptase inhibitors (NNRTIs)resistance-related gene mutations among the AIDS patients with virological suppression failure in Guangdong Province 2015.Methods Plasma samples from AIDS patients receiving highly active antiretroviral therapy for more than one year with viral loads > 1000 copies/mL from Guangdong province (except Shenzhen)were collected from January to December 2015.Total 612 HIV-1 gene fragments were amplified from plasma samples using self-developed lab method.Sub-genotypes were determined by phylogenetic tree according to the sequences,NNRTIs resistance-related mutations were determined in Stanford University HIV-1 Drug Resistance Database. The NNRTIs-resistance, the relationships of NNRTIs resistance-related mutations with baseline CD4 +T lymphocyte counts,transmission routes,antiviral regimens and HIV-1 genotypes were analyzed.SPSS 17.0 software was used to analyze the data.Results In 612 patients with virological suppression failure,the main NNRTIs resistance-related mutations were K103 (26.80%),Y181 (14.71 %),V179 (13.73%),G190 (11 .44%) and V106 (10.62%).The susceptibility rate of 310 patients (50.65%)to NNRTIs had changed,the highly resistant rate to nevirapine was 49.51 %,which was higher than that of efavirenz (43.14%),etravirine (5.56%) and rilpivirine (12.25%),respectively,and the differences were statistically significant (χ2 =5.00,296.3 and 198.0,all P <0.05).The incidence rate of drug resistance in patients with baseline CD4 +T lymphocyte counts >200 cells/μL was lower than that in those with baseline CD4 +T lymphocyte counts <200 cells/μL (χ2 =17.93,P <0.01 );the incidence rate of drug resistance was lower in intravenous drug abusers than that of sexually transmitted patients (χ2 =44.21 ,P <0.01 );while the incidence of drug resistance in patients receiving NVP-containing regimens was higher than that in those receiving EFV-containing regimens (χ2 =8.93,P <0.01 ),and the incidence rate was higher in patients with CRF01 _AE than that in those with CRF07_BC and CRF08 _BC (χ2 =8.46 and 8.47,P <0.01 ).Conclusions The results suggest that compliance education and follow-up should be strengthened in patients with high baseline CD4 +T lymphocyte counts and intravenous drug users,and patients with liver diseases should avoid using drugs containing NVP regimens.

OBJECTIVE:To explore the clinical efficacy and safety of oxymatrine membrane,rhEGF gel respectively com-bined with Yunnan baiyao for postoperative cervical wound after LEEP. METHODS:300 patients with cervical intraepithelial neo-plasia(CIN)Ⅱ and Ⅲ were divided into group A,B,C(100 cases in each group)based on CIN grading and stratification and random sampling in each stratification. After conventional LEEP,patients in group A were cleaned the wound by 0.9% Sodium chloride injection,spraying Yunnan baiyao powder,once only after surgery;patients in group B were additionally given recombi-nant human epidermal growth factor (rhEGF) on the basis of group A,once every week after surgery,for 3 weeks;patients in group C were additionally given oxymatrine membrane on the basis of group A,1 tablet every evening after surgery,for 2 weeks. Postoperative bleeding,bleeding duration,rebleeding and duration after postoperative bleeding stopped,postoperative drainage du-ration,the incidence of adverse reactions in 3 groups were observed. RESULTS:The patients of postoperative bleeding,bleeding time ≥7 d and rebleeding after stopping bleeding in group B were significantly lower than group A;the incidence of bleeding time ≥7 d in group C was significantly lower than group A,the proporition of postoperative drainage duration for less than 7 d was significantly higher than group A,for 8-13 d was significantly less than group A;early wound healing rate in group B and group C were significantly better than group A,with statistical significances (P<0.05). There were no significant differences in above-mentioned indexes in group B and group C(P>0.05),and there were no obvious adverse reactions in 3 groups. CONCLU-SIONS:Oxymatrine membrane and rhEGF gel respectively combined with Yunnan baiyao have better healing than Yunnan baiyao alone,do not increase the incidence of adverse reactions,while there is no significant difference in oxymatrine membrane and rhEGF gel.

Objective To detect the change of hepatitis C virus (HCV)RNA in the peripheral blood mononuclear cells (PBMC)and serum of patients with chronic hepatitis C (CHC)during treatment with peg-interferon α-2a (Peg IFNα-2a)plus ribavirin (RBV),and to analyze the clinical significance of HCV RNA detection in PBMC.Methods The peripheral blood samples of 20 CHC patients who visited Department of Infectious Diseases in Guangzhou No.8 People′s Hospital from June 2013 to December 2014,were collected during treatment with Peg IFNα-2a+RBV at different time points (week 0,2,4, 12,24,36 and 48).Serum and PBMC were separated.Accurate fluorescence quantification assay (Cobas TaqMan real time polymerase chain reaction[PCR])was used to detect HCV RNA level in serum,while real-time PCR and nest-PCR were applied to detect HCV RNA in PBMC.Categorical data were analyzed byχ2 test.Results Accurate fluorescence quantification of serum HCV RNA showed that HCV RNA level decline rapidly after treatment (F = 148.06,P < 0.01 ),and 18 patients achieved HCV RNA undetectable at week 12 of treatment.The positive rate of nest-PCR was higher than real-time PCR (all P <0.01).Comparison of HCV RNA levels in serum and PBMC from 20 cases found that,the clearance rate of HCV RNA in PBMC was postponed.Two patients whose HCV RNA in PBMC kept detectable relapsed at week 24 after end of treatment.Conclusions HCV RNA can be detected in PBMC of CHC patients and the positive rate of nest-PCR is higher than real-time PCR.Antiviral therapy is effective on HCV both inside and outside PBMC,but the clearance rate of HCV RNA in PBMC is postponed compared with that in serum.Slow clearance of HCV in PBMC may be a risk factor for relapse after end of treatment.

Objective To investigate the relationship between HBV mutations in the precore (PC)/core promoter region and the liver histological changes in HBeAg negative CHB patients. Method A total of 71 HBeAg negative CHB patients with liver biopsy from April 2012 to Dec 2013 were enrolled. DNA was extracted from blood serum, then the HBV S gene and PC/core promoter region were amplified by semi-nested PCR and sequenced. The relationship between significant liver histological changes and viral factors were analyzed by Logistic regression analysis. Results The incidence of significant necroinflammation (15.8% vs. 27.3%, χ2 =1.398, P = 0.237) and significant fibrosis (71.1% vs. 84.4%, χ2= 1.926, P = 0.165) were found to be similar between patients infected with HBV genotype B and genotype C . By Logistic regression analysis including risk factors of age, sex, HBV genotype and mutations (T1753V,A1762T/G1764A,A1846T and G1896A), the A1762T/G1764A mutation in HBV associated with significant necroinflammation (OR = 4.296, P = 0.037), while factors of age, sex, genotype and other mutation were not associated with significant liver histological changes. (all P > 0.05). Conclusion Mutation in PC/core promoter region of HBV may act as a marker to evaluate the liver histological changes.

Objective To detect the relationship of pathological grade and stage of bladder cancer with common chromosomal aberrations of urine exfoliated cells by FISH. Methods A total of 99 urine samples were detected by FISH with probes of chromosomes 3,7,17 and 9p21 to collect pathological grade and stage information of bladder tumor tissues. Results (1) The aberrations of chromosome 3 and 17 had significant correlation with pathological grade and stage (P<0.05) but that of chromosome 7 had no correlation with pathological stage (P>0.05), but had correlation with grade (P < 0.05). The aberration of 9p21 had no correlations with pathological grade or stage (P > 0.05). (2)The polysomic chromosomal aberrations of chromosomes 3, 7 and 17 assessed correlated with high-grade and high-stage bladder carcinoma. The 9p21 deletion was found at a significant frequency in low-grade and low-stage lesions, when, 9p21 amplification was found at a significant frequency in high-grade and high-stage lesions. Conclusions Aberrations of the four chromosomes, especially polysomic chromosomal aberrations of the bladder cancer cases could present a possible trend toward greater chromosome increased with tumor grades and progressive stages of invasion.

Objective To investigarte the immune status in the patients with dengue fever .Methods The flow cytometry was used to detect the T lymphocytes in the patients with dengue fever for analyzing their immune status .Results Compared with the reference range in the healthy individuals ,it was found that the T lymphocyte proportion in the patients with dengue fever was sig‐nificantly reduced compared with the healthy individuals .The percentages of CD3+CD4+ and CD3+CD8+ lymphocytes were (36 .54 ± 9 .78)% and (17 .7 ± 10 .01)% respectively ,which had statistical difference compared with the control group(P<0 .05) ,CD3+CD4+ lymphocyte count was (49 .98 ± 240 .2)cells/μL ,the difference was statistically significant (P<0 .05) ,CD3+ CD8+ lympho‐cytes count was (380 .9 ± 364 .6)cells/μL ,the difference was statistically significant(P<0 .05) .Conclusion The immune status in the patients with dengue fever is abnormal ,T lymphocyte percentage is significantly reduced compared with the healthy individuals .

Objective To investigate a non-invative , sensitive and specific method to diagnose bladder cancer, and evaluate the value of the combination of FISH with NMP22 in the diagnosis of bladder cancer. Methods Urine from 68 patients suspected suffering from bladder cancer were used by FISH, NMP22 expression and cytologi-cal examination, respectively. The results of above detections were compared to the subsequent histopathology as-say to analyze the specicficity, sensitivity of diagnosis for bladder cancer. Results The sensitivity of FISH, NMP22 expression and the cytological examination was 81.4%,86.0% and 39.5% respectively, and the specifici-ty of FISH, NMP22 expression and the cytological examination was 84.0%, 72.0% and 96.0%, respectively. The sensitivity and specificity of the combination of FISH with NMP22 was 79.1% and 92.0%. Conclusions The combined diagnosis of FISH and NMP22 for bladder cancer showed greater sensitivity than that of the conventional cytology, with similar specificity as the conventional cytology. The combination of FISH with NMP22 test shows more value for the diagnosis of bladder cancer than any single assay.

Objective To investigate the characteristics of V3 loop amino acid sequences of human immunodeficiency virus type 1 (HIV-1) quasi-species in long-term non-progressors (LTNP)infected with HIV. Methods End-point limiting dilution polymerase chain reaction (EPLD PCR) was used to amplify the env gene c2-v3-c3 region of single HIV-1 provirus from five LTNPs at sequential time points. The PCR products were then sequenced and the amino acid sequences of V3 loop were analyzed by sequence confirm analysis technology. Results The results showed that there were one to ten kinds of polymorphisms in the V3 region of HIV-1 quasi-species which were found from the serial samples of the five LTNP. However, the sequences of the predominant strains were either completely consistent or at most changed at one or two residues in the serial samples of individual patient. The tetramer compositions of the tip of V3 loop were consistent in each patient. It was GPGR in four patients and GPGK in one patient. It was speculated the co-receptor of HIV-1 was CC chemokine receptor (CCR)-5 based on the amino acids at the residue 11 and residue 25 of V3 loop and the net charge of V3 loop. Conclusions There are various polymorphisms at the HIV V3 loop in LTNP. However, the tetramer composition of the tip part of V3 loop is stable. The LTNP are very likely infected with non-syncytium inducing (NSI) strain.