Development of precocious puberty in children: Surmised medicinal plant treatment.

Precocious puberty in children is one of the common endocrine diseases in paediatrics. Epidemiological surveys have shown that the number of children with precocious puberty has significantly increased globally. Precocious puberty negatively affects the physical and mental health of children and may increase the risk of hypertension, diabetes, obesity and infertility in adulthood. Therefore, the initiating factors of adolescence have become core issues in the study of sexual development in children. Owing to developments in molecular genetics, many studies have been able to show that precocious puberty is mostly resulted in autosomal inheritance. For instance, makorin ring finger protein 3 gene (MKRN3) may be implicated in familial CPP. Gonadotropin-releasing hormone agonist (GnRHa) is the gold standard for treatment, but its safety still requires long-term evaluation and management. Traditional medicinal plants have been used in clinical treatments and in exploring novel treatment methods. From the collected datas, in Asia, traditional Chinese medicine treatment is based on the principles of nourishing Yin, lowering fire and draining fire from the liver to help precocious children and alleviate or delay the onset of precocious puberty by medicinal plants such as Anemarrhena asphodeloides Bge., Phellodendron amurense Rupr., Rehmannia glutinosa and Poria cocos Wolf. They play an important role in exploring the pharmacological mechanisms of precocious puberty treatment effects and drug development. Therefore, by elucidating the occurrence and development of precocious puberty, this review provides novel and valuable insights of paediatric endocrine therapy accessing the published researches on the effectiveness of traditional herbal medicine in the treatment of precocious puberty and its therapeutic mechanisms.

miR-450a exerts oncosuppressive effects in breast carcinoma by targeting CREB1.

Emerging evidence greatly implicates that microRNA-450a (miR-450a) plays an essential role in cancer pathobiology. While the pathological role of miR-450a in breast carcinogenesis remains enigmatic. Herein, we showed that miR-450a was lowly expressed in breast cancer cell lines compared with normal, and low miR-450a expression was associated with poor survival in patients with breast cancer. We revealed that miR-450a mimic transfected breast cancer cells (T47D and BT474) exhibited attenuated capacities of proliferation, migration, and invasion in vitro, and miR-450a suppressed T47D cell growth in a xenograft tumor model. Mechanistically, cAMP response element-binding protein 1 (CREB1) was negatively targeted by miR-450a, and CREB1 deletion mimicked the effects of miR-450a mimic treatment. Bioinformatics analysis further revealed that elevated expression of CREB1 correlated with poor prognosis in patients with breast cancer and miR-450a level was negatively correlated with CREB1 level in breast cancer. Additionally, miR-450a inhibited the phosphorylation of phosphatidylinositol 3-kinase/V-akt murine thymoma viral oncogene homolog (PI3K/AKT) and the activities of matrix metalloproteinase-2/9 (MMP-2/9). The following rescue assay indicated that CREB1 was implicated in the anti-tumoral effect of mR-450a in breast carcinoma. All these observations disclosed that miR-450a negatively regulates the growth and metastatic property of breast carcinoma cells.

C6orf120 gene knockout in rats mitigates concanavalin A­induced autoimmune hepatitis via regulating NKT cells.

OBJECTIVE: To elucidate the role of the functional unknown gene C6orf120 in the pathogenesis of AIH and its mechanism of action, using C6orf120 knockout rats. METHODS: An autoimmune hepatitis model was established with 35 mg/kg intravenous injection of concanavalin A (Con A) in C6orf120-knockout (C6orf120-/-) and wild-type (WT) rats. Rats were sacrificed after administering Con A for 0, 12, and 24 h. The peripheral blood, liver, spleen, and mesenteric lymph nodes were collected for follow-up studies. RESULTS: C6orf120 knockout significantly decreased the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and improved the histological damage in Con A-induced autoimmune liver injury.Loss of C6orf120 function significantly increased the frequency of CD3+ CD161+ NKT cells in the peripheral blood, liver, and spleen; downregulated the expression of CD314 (NKG2D) in the liver, spleen, and mesenteric lymph nodes; reduced the expression of inflammatory cytokines and chemokines; and suppressed the mRNA and protein expression of Fas and FasL in the liver. Additionally, C6orf120 knockout significantly downregulated the expression of p-JAK1, p-JAK2, p-STAT1, and p-STAT3 in liver tissue. CONCLUSION: The protective effect of C6orf120 knockout against Con A-induced hepatitis may be due to the inhibition of NKT cell activation, restriction of cytokine and chemokine activities, inhibition of JAK-STAT and Fas/FasL signaling pathway activation, and reduction in liver inflammation and hepatocyte apoptosis.

Prevention and control of HIV/AIDS in China: lessons from the past three decades.

ABSTRACT: In the past 37 years, human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) has undergone various major transmission routes in China, with the world most complex co-circulating HIV-1 subtypes, even the prevalence is still low. In response to the first epidemic outbreak of HIV in injecting drug users and the second one by illegal commercial blood collection, China issued the Anti-Drug Law and launched the Blood Donation Act and nationwide nucleic acid testing, which has avoided 98,232 to 211,200 estimated infections and almost ended the blood product-related infection. China has been providing free antiretroviral therapy (ART) since 2003, which covered >80% of the identified patients and achieved a viral suppression rate of 91%. To bend the curve of increasing the disease burden of HIV and finally end the epidemic, China should consider constraining HIV spread through sexual transmission, narrowing the gaps in identifying HIV cases, and the long-term effectiveness and safety of ART in the future.

Disparity of human immunodeficiency virus incidence and drug resistance in college student, non-student youth and older men who have sex with men: a cross-sectional study from seven major cities of China.

BACKGROUND: Human immunodeficiency virus (HIV) prevalence among student men who have sex with men (MSM) in college is more than 5.0% and keeps on increasing in China. This study aims to clarify the proportion of HIV recent infection, its propeller and the source among college student MSM. METHODS: We conducted a multicenter cross-sectional study in seven major Chinese cities during 2012-2013. HIV recent infections (≤ 168 days) and incidence was measured and estimated by BED HIV-1 capture enzyme immunoassay (BED-CEIA) testing strategy. HIV-related behaviors and transmitted drug resistance (TDR) were investigated and compared between the college student MSM, <25-year-old non-student youth MSM (NSYM), and ≥25-year-old non-student non-youth MSM (NSNYM), using structured survey, and analyses of drug resistance. RESULTS: Overall, 4,496 (4496/4526, 99.3%) were eligible for enrollment, comprising 565 college student MSM, 1,094 NSYM, and 2,837 NSNYM. The proportion of HIV recent infection were 70.3% (26/37), 50.8% (65/128) and 35.1% (95/271), the HIV incidence rate were 10.0 (95% CI: 6.2-13.9)/100PY, 12.9 (95% CI: 9.8-16.1)/100PY, 6.8 (95% CI: 5.4-8.2)/100PY, and TDR prevalence were 7.4% (2/27), 2.0%, (2/98) and 4.9% (11/226), among student MSM, NSYM, and NSNYM, respectively. Among HIV positive student MSM with age< 21-year-old, the proportion of HIV recent infection is 90.9% (10/11). Factors independently associated with HIV recent infection in student MSM was usage of recreational drug in the past 6 months (AOR: 2.5; 95% CI: 1.0-5.8). CONCLUSIONS: College student MSM had higher proportion of HIV recent infection and TDR than the youth and older MSM in China during 2012-2013. The HIV infections were more likely to happen during the early year of college life among student MSM.

Efficacy and cost-effectiveness of early antiretroviral therapy and partners' pre-exposure prophylaxis among men who have sex with men in Shenyang, China: a prospective cohort and costing study.

BACKGROUND: Biomedical interventions such as antiretroviral therapy (ART) and pre-exposure prophylaxis (PrEP) are highly effective for prevention of human immunodeficiency virus (HIV) infection. However, China has not released national PrEP guidelines, and HIV incidence among men who have sex with men (MSM) is unchanged despite substantial scale-up of ART. We evaluated reductions in HIV transmission that may be achieved through early initiation of ART plus partners' PrEP. METHODS: Six intervention scenarios were evaluated in terms of their impact on HIV transmission and their cost-effectiveness for 36 months post-infection. Three scenarios were based on observed data: non-ART, standard-ART, and early-ART. Another three scenarios were based on observed and hypothetical data: non-ART plus partners' PrEP, standard-ART plus partners' PrEP, and early-ART plus partners' PrEP. The number of onward transmissions was calculated according to viral load and self-reported sexual behaviors, and calibrated by the prevalence and incidence of HIV among Chinese MSM. Cost-effectiveness outcomes were quality-adjusted life-years (QALYs) and cost-utility ratio (CUR). RESULTS: The estimated number of onward transmissions by every 100 HIV-positive cases 36 months post-infection was 41.83 (95% credible interval: 30.75-57.69) in the non-ART scenario, 7.95 (5.85-10.95) in the early-ART scenario, and 0.79 (0.58-1.09) in the early-ART plus partners' PrEP scenario. Compared with non-ART, the early-ART and early-ART plus partners' PrEP scenarios were associated with an 81.0 and 98.1% reduction in HIV transmission, and had a CUR of $12,864/QALY and $16,817/QALY, respectively. CONCLUSIONS: Integrated delivery of early ART and sexual partners' PrEP could nearly eliminate HIV transmission and reduce costs during the first 36 months of HIV infection. Our results suggest a feasible and cost-effective strategy for reversing the HIV epidemic among MSM in China.

Protective effects of Astragalin on spermatogenesis in streptozotocin-induced diabetes in male mice by improving antioxidant activity and inhibiting inflammation.

Spermatogenic dysfunction is a common complication in men with diabetes and is the most important manifestation of diabetes-related male reproduction damage. Astragalin (AG) is one of the main flavonoids from Cuscuta chinensis, which has rich pharmacological activities. This study aimed to establish whether AG may contribute to the recovery from spermatogenic dysfunction. AG (3.3, 10 and 30 mg/kg) and Clomiphene (5 mg/kg) were orally administered to streptozotocin-induced diabetic male mice for 8 weeks. After the experiments performed, reproductive organs, sperm parameters and histomorphological changes were analysed. Antioxidant and anti-inflammatory capacity were estimated in testicular tissues. The results revealed that AG significantly improved the reproductive organs, sperm parameters and testicular morphology to different degrees in diabetic mice. Nitric oxide (NO) and malondialdehyde (MDA) levels were significantly reduced, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), markedly increased in the testicular tissue after AG was administered. Interestingly, AG also downregulated the protein expressions of tumour necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) in testes. In conclusion, AG is a potential beneficial agent to protect diabetic-induced spermatogenic dysfunction in male mice by increasing antioxidant enzymes activities and inhibiting inflammation.

Lycium barbarum polysaccharide attenuates diabetic testicular dysfunction via inhibition of the PI3K/Akt pathway-mediated abnormal autophagy in male mice.

Testicular dysfunction is one of the serious secondary complications in diabetes. Lycium barbarum polysaccharide (LBP) has long been considered to possess a wide range of beneficial properties including antiaging, anticancer and reproductive-enhancing. Abnormal autophagy was reported to play a significant role in accelerating diabetic reproductive injury. However, the autophagy regulation mechanism of LBP on diabetic testicular dysfunction is incompletely understood. We investigate the protective effects of LBP on diabetic testicular dysfunction and its underlying mechanism with different approaches. Protective effects of LBP (40 mg/kg) on testicular functions were assessed through the use of sperm parameters, testosterone levels and hematoxylin and eosin staining. Antioxidant capacity and serum malondialdehyde levels were determined using assay kits. Immune intensity of Beclin-1 and LC3I in testes was detected by immunofluorescence staining. Western blot analysis was used to detect expressions of p-PI3K, Akt, p-Akt, Beclin-1, LC3I and LC3II proteins. Q-PCR was used to evaluate Beclin-1 and LC3I mRNA expressions in testis. Administration of LBP (40 mg/kg) considerably recovered testicular function, obviously improved testicular histopathologic structure and significantly increased antioxidant enzyme activities. Immunofluorescence staining showed that immune intensity of Beclin-1 and LC3I significantly decreased in the LBP 40 mg/kg group. The results of Q-PCR and western blot analysis showed that LBP 40 mg/kg significantly downregulated Beclin-1 and LC3I protein expressions upregulated p-PI3K and p-Akt protein expressions and decreased Beclin-1 and LC3I mRNA expressions compared with diabetic mice. In conclusion, inhibition of PI3K/Akt pathway-mediated testicular excessive autophagy may be a target for protective effects of LBP on diabetic testicular dysfunction.

Diabetes associated with male reproductive system damages: Onset of presentation, pathophysiological mechanisms and drug intervention.

Diabetes mellitus (DM) is a major health problem that affects patients' quality of life quality throughout the world due to its many complications. Reproductive dysfunction is one of the major secondary complications in both diabetic animals and human beings. Furthermore, DM has recently broken the age barrier and has been heavily diagnosed in children and young persons of reproductive age. In the past few years, many studies on DM in male reproductive functions in both diabetic men and experimental diabetic animals have been published. It is recognized that sustained hyperglycemia, which impairs reproductive function in diabetic men, is at risk of developing. DM harmfully affects male reproductive functions in multiple areas; these may include spermatogenesis, sperm maturation, fertility capability, penile erection, and ejaculation. Traditional medicine and folklore worldwide have used numerous medicinal plants to manage the diabetic reproductive dysfunction because bioactive phyto-constituents are affluent in many places. Unfortunately, the exact reasons for diabetic male reproductive dysfunction are not completely understood and currently there are no treatments in reproductive medicine specifically for such lesions. The aim of this review is to summarize current research findings of DM on reproductive functions, to elaborate the underlying mechanisms related to these diseases via in vivo and in vitro studies, and to describe the ameliorative effects of medicinal plants or their products. The review findings provide a systematic understanding of DM on the reproductive functions and lay the theoretical foundation for developing the direction of reproductive medicine.

Commercial sex and risk of HIV, syphilis, and herpes simplex virus-2 among men who have sex with men in six Chinese cities.

BACKGROUND: Men who have sex with men (MSM) are at high risk of HIV and sexually transmitted infections (STIs) in China and globally. Engaging in commercial sex put them at even greater risk. This study estimated the prevalence of HIV/STIs among three subgroups of MSM: MSM who sold sex (MSM-selling), MSM who bought sex (MSM-buying), and non-commercial MSM (NC-MSM) and evaluated the relationship between commercial sex and HIV/STIs. METHODS: We conducted a cross-sectional survey among MSM in six Chinese cities (Shenyang, Ji'nan, Changsha, Zhengzhou, Nanjing, and Kunming) from 2012 to 2013. Data on socio-demographics and sexual behaviors were collected. Serological tests were conducted to detect HIV, syphilis, and human simplex virus type 2 (HSV-2). RESULTS: Of 3717 MSM, 6.8% were engaged in commercial sex. The overall prevalence of HIV, syphilis and HSV-2 infections was 11.1, 8.8 and 12.1%, respectively. MSM-selling had higher prevalence of HIV (13.4%), syphilis (12.1%) and HSV-2 (17.9%) than NC-MSM (10.9, 8.7 and 11.9% for HIV, syphilis and HSV-2, respectively), though the differences are not statistically significant. Among MSM-selling, HIV prevalence was significantly higher for those who found sex partners via Internet than those did not (19.4% vs. 8.1%, P = 0.04). Compared to NC-MSM, MSM-selling were more likely to use recreation drugs (59.3% vs. 26.3%), have unprotected anal intercourse (77.9% vs. 61.7%), and have ≥10 male sex partners (46.2% vs. 6.2%) in the past 6 months (each P < 0.05). CONCLUSIONS: All three subgroups of MSM in six large Chinese cities have high prevalence of HIV/STIs. Those who sell sex only have a particularly high risk of acquiring and transmitting disease, and therefore, they should be considered as a priority group in HIV/STIs surveillance and intervention programs.

High HIV incidence epidemic among men who have sex with men in china: results from a multi-site cross-sectional study.

BACKGROUND: Recent upsurge of new HIV infections among men who have sex with men (MSM) is a major concern in China. Paucity of national-level information regarding the burden and predictors of this progressive epidemic of new infections called for a multi-centric, timely and comprehensive investigation. METHODS: Mixed methods were used to recruit MSM from seven cities in China between 2012 and 2013. Recent and established HIV infections were estimated by Western Blot and BED HIV-1 capture enzyme immunoassay. Syphilis and herpes simplex virus-2 (HSV-2) were also tested. RESULTS: A total of 4496 eligible MSM were recruited. The majority was aged ≤35 years (77.5 %), migrants (60.3 %), never married (69.8 %), and played receptive role in anal sex (70.5 %). The HIV prevalence was 9.9 %, and 41.9 % were recently infected, with sensitivity/specificity adjusted HIV incidence of 8.9 (95 % CI: 7.6-10.2)/100 Person-Years. The prevalence of history HSV-2 and syphilis were 12.5 % and 8.5 %, respectively. Recent HIV infection was associated with having multiple male partners (aOR = 1.4, 95 % CI 1.1-1.9), recreational drug use (aOR = 2.2, 95 % CI 1.6-3.0), anal bleeding (aOR = 2.1, 95 % CI 1.4-3.0), syphilis infection (aOR = 2.8, 95 % CI 1.9-4.3) and history HSV-2 infection (aOR = 2.3, 95 % CI 1.5-3.3). CONCLUSION: High rate of recent HIV infection is potentially resulting in progressive deterioration of the overall HIV epidemic among MSM in China. Targeted interventions to address high-risk MSM including those having multiple partners, history of recreational drug use and syphilis or HSV-2 infection seemed to be the need of the hour.

Characterization of a New S8 serine Protease from Marine Sedimentary Photobacterium sp. A5-7 and the Function of Its Protease-Associated Domain.

Bacterial extracellular proteases are important for bacterial nutrition and marine sedimentary organic nitrogen degradation. However, only a few proteases from marine sedimentary bacteria have been characterized. Some subtilases have a protease-associated (PA) domain inserted in the catalytic domain. Although structural analysis and deletion mutation suggests that the PA domain in subtilases is involved in substrate binding, direct evidence to support this function is still absent. Here, a protease, P57, secreted by Photobacterium sp. A5-7 isolated from marine sediment was characterized. P57 could hydrolyze casein, gelatin and collagen. It showed the highest activity at 40°C and pH 8.0. P57 is a new subtilase, with 63% sequence identity to the closest characterized protease. Mature P57 contains a catalytic domain and an inserted PA domain. The recombinant PA domain from P57 was shown to have collagen-binding ability, and Phe349 and Tyr432 were revealed to be key residues for collagen binding in the PA domain. This study first shows direct evidence that the PA domain of a subtilase can bind substrate, which provides a better understanding of the function of the PA domain of subtilases and bacterial extracellular proteases from marine sediment.

Internet-Based Sex-Seeking Behavior Promotes HIV Infection Risk: A 6-Year Serial Cross-Sectional Survey to MSM in Shenyang, China.

HIV prevalence is still rapidly increasing among Chinese men who have sex with men (MSM). The Internet also makes it easier for MSM to have casual partners. This study aims to evaluate the trend of Internet-based sex-seeking behavior of MSM and its impact on HIV prevalence, the distribution of HIV subtype strains, and transmitted drug resistance rates. A serial cross-sectional study was conducted from 2009 to 2014. Of the 1,981 MSM, 50.5% (1,000/1,981) mainly sought homosexual partners through the Internet (Internet-based MSM, IBM). The proportion of IBM among total MSM subjects increased from 43.3% to 61.5% (p < 0.001). HIV prevalence of IBM increased from 5.7% to 20.7%, while that of non-Internet-based MSM (NIBM) increased from 7.0% to 14.7%. A relative higher proportion of NIBM were infected with HIV CRF01_AE subtype than IBM (79.5% versus 72.2%, p = 0.52). Multivariable analysis found IBM had a significantly higher HIV prevalence than NIBM (13.2% versus 10.5%, aOR = 1.4, 95% CI [1.0-1.9]). Being a migrant non-Shenyang resident MSM (aOR = 1.9, 95% CI [1.3-2.9]) and occasionally/never using condoms with casual homosexual partners (aOR = 1.7, 95% CI [1.1-2.6]) were two distinct risk factors for HIV infection in IBM. More efforts should be targeted towards developing interventions aimed at IBM, particularly migrant MSM and who engage in UAI with casual homosexual partners.

Diversity of cultivable protease-producing bacteria in sediments of Jiaozhou Bay, China.

Although protease-producing bacteria are key players in the degradation of organic nitrogen and essential for the nitrogen recycling in marine sediments, diversity of both these bacteria and their extracellular proteases is still largely unknown. This study investigated the diversity of the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the eutrophied Jiaozhou Bay, China through phylogenetic analysis and protease inhibitor tests. The abundance of the cultivable protease-producing bacteria was up to 10(4) cells/g in all six sediment samples. The cultivated protease-producing bacteria mostly belonged to the phyla Proteobacteria and Firmicutes with the predominant genera being Photobacterium (39.4%), Bacillus (25.8%), and Vibrio (19.7%). Protease inhibitor tests revealed that extracellular proteases secreted by the bacteria were mainly serine proteases and/or metalloproteases with relatively low proportions of cysteine proteases. This study represents the first comprehensive analysis on the diversity of protease-producing bacteria and their extracellular proteases in sediments of a eutrophic bay.

The biological characteristics of predominant strains of HIV-1 genotype: modeling of HIV-1 infection among men who have sex with men.

To investigate the molecular subtypes of prevalent HIV-1 strains and characterize the genetics of dominant strains among men who have sex with men. Molecular epidemiology surveys in this study concentrated on the prevalent HIV-1 strains in Liaoning province by year. 229 adult patients infected with HIV-1 and part of a high-risk group of men who have sex with men were recruited. Reverse transcription and nested PCR amplification were performed. Sequencing reactions were conducted and edited, followed by codon-based alignment. NJ phylogenetic tree analyses detected two distinct CRF01_AE phylogenetic clusters, designated clusters 1 and 2. Clusters 1 and 2 accounted for 12.8% and 84.2% of sequences in the pol gene and 17.6% and 73.1% of sequences in the env gene, respectively. Another six samples were distributed on other phylogenetic clusters. Cluster 1 increased significantly from 5.6% to 20.0%, but cluster 2 decreased from 87.5% to 80.0%. Genetic distance analysis indicated that CRF01_AE cluster 1 in Liaoning was homologous to epidemic CRF01_AE strains, but CRF01_AE cluster 2 was different from other scattered strains. Additionally, significant differences were found in tetra-peptide motifs at the tip of V3 loop between cluster 1 and 2; however, differences in coreceptor usage were not detected. This study shows that subtype CRF01_AE strain may be the most prevalent epidemic strain in the men who have sex with men. Genetic characteristics of the subtype CRF01_AE cluster strain in Liaoning showed homology to the prevalent strains of men who have sex with men in other parts of China.

[Study on the rates of infection and spontaneous clearance on HCV among HIV-infected men who have sex with men in China].

OBJECTIVE: The aim of this study was to understand the rates of infection and spontaneous clearance on hepatitis C virus (HCV) among HIV-infected men who have sex with men (MSM) in China. METHODS: HIV-infected MSM from Shenyang, Beijing, Changsha and Kunming were selected to take the HCV antibody and HVC-RNA tests, and then classified into 'under recovery (RNA-/Ab+) ' or being chronic (RNA+/Ab+). Information on demography, CD4(+) T cell, HIV viral load and liver biochemical indicators were also collected. RESULTS: In 513 HIV-infected MSM, the positive rate of HCV antibody was 1.94% (10/513) and the positive rate of HVC-RNA was 0.77% (4/513). Five patients showed clearance of HVC-RNA, spontaneously. These 5 patients had low levels of both alanine transaminase (ALT) and aspartate aminotransferase (AST), and with high level of CD4(+) T cells, when compared to those who were not able to clear the HVC-RNA. CONCLUSION: Currently, the HCV infection rate was still at quite low level among the HIV-infected MSM in China. Dynamic of HIV/HCV co-infection should be monitored so as to develop suitable strategy on prevention and treatment among this population, in China.

[Study on the association between genetic variation of gag gene and HLA restricted CTL response in HIV-1 B' infected Chinese people].

OBJECTIVE: To identify the genetic variation of Gag gene in HIV-1 B' subtype infected Chinese people and to evaluate the association between the positive selection sites and HLA restricted CTL response. METHODS: We downloaded all HIV-1 B' subtype gag gene sampled from China from the LANL HIV database. The sequences were grouped by years of collection and the mean intra-subtype diversity and divergence were examined by MEGA 5.0 software. The sites under positive selection were estimated by using Hyphy software. Then analyzed the association between positively selected sites and HLA restricted CTL epitopes. RESULTS: A set of 122 B' subtype HIV-1 gag sequences were collected from Henan, Hubei, Liaoning and Yunnan provinces, grouped by sampling year as < 2001, 2001 - 2003, 2004 - 2006 and 2007 - 2010. The mean intra-subtype diversity and divergence increased over time (r(1) = 0.497, P1 < 0.001; r(2) = 0.593, P2 < 0.001). This analysis revealed 34 positively selected sites and 97.1% of these sites were located within 47 known CTL epitopes. The most frequent HLA was HLA-A*02 (21.6%) followed by A*11(9.8%), B*35(9.8%) and B*57(5.9%). The percentages of HLA-A/B/C limited CTL epitopes were 43.1%, 49.0% and 7.9%, respectively. CONCLUSION: The mean intra-subtype diversity and divergence of B' subtype gag sequences increased over time. B' subtype HIV-1 virus has accumulated adaptations to HLA restricted CTL responses in HIV-1 infected Chinese.

[Association between amino acid mutations in Gag protein and human leukocyte antigen class I alleles in human immunodeficiency virus-1 B' infected Han Chinese people].

OBJECTIVE: To explore the human leukocyte antigen (HLA)-associated mutations in Gag protein of B' clade (human immunodeficiency virus-1) HIV-1 infected Han Chinese people and evaluate the impact of HLA associated Gag mutations on the disease progression of HIV infection. METHODS: A total of 95 B' clade HIV-1 infected Han Chinese cases were recruited. The gag sequences were amplified from viral RNA and sequenced directly. HLA-I genotypes were detected with the assay of polymerase chain reaction-sequence specific primer (PCR-SSP). HLA-associated mutations were identified and the relationships between HLA-associated mutations and CD4+ T cell counts or plasma viral loads analyzed. RESULTS: Forty-seven kinds of mutations at 28 sites (15, 18, 26, 30, 34, 46, 62, 67, 81, 84, 90, 102, 118, 121, 122, 125, 146, 147, 173, 176, 252, 357, 374, 376, 437, 470, 471, 478) of Gag protein were significantly associated with particular HLA class I allelotypes (P < 0.05). Among which, 9 sites (26, 30, 81, 84, 125, 146, 147, 357, 437) were located within 13 known cytotoxic T-lymphocyte (CTL) epitopes or flanking regions. The number of HLA-associated mutations was significantly associated with both CD4 T cell counts (r = -0.318, P = 0.002) and viral loads (r = 0.360, P = 0.003). CONCLUSION: HLA-associated mutations may have a significant impact on HIV disease progression in B' clade HIV-1 infected Han Chinese population.

[Coreceptor usage of human immunodeficiency virus-1 in primary infection through sexual contact transmission].

OBJECTIVE: To evaluate the coreceptor usage characters and evolution of human immunodeficiency virus-1 (HIV-1) primary infection. METHODS: A total of 12 HIV-1 primary infection cases were recruited from a high-risk population for this prospective cohort trial. The blood samples were collected at the earliest time after infection and after a viral setpoint respectively. RNA was extracted from plasma. The env genes were amplified through reverse transcription-PCR (RT-PCR) and nested PCR. And the env fragments were ligated to T vector and then sequenced. The pattern of coreceptor usage was predicted with five different online tools. RESULTS: Among them, 11 were predicted as CCR5 virus transmission and 1 as CXCR4 virus transmission. The viruses in 1 case were detected as a switch of coreceptor usage from CCR5 to CXCR4. The consistency of five different prediction rules was 88.3%. CONCLUSION: Although the CCR5 usage virus is predominant in primary infection through sexual transmission, CXCR4 usage virus can also be detected. A switch of coreceptor usage may happen within the first year post-infection. Adopting multiple rules may improve the efficiency and validity for the prediction of coreceptor usage based on genotypes.

Association of variations of NAb 2F5 and 4E10 epitopes and disease progression in Chinese antiretroviral treatment-naïve patients infected with HIV-1 clade B'.

BACKGROUND: Studies on human immunodeficiency virus type 1 (HIV-1) vaccines have recently focused on targeting the conserved neutralizing epitopes 2F5 and 4E10, and hence it is important to understand the extent of mutations in these two viral epitopes. Here, we investigated the amino acid mutations in epitopes of 2F5 (ELDKWA, HIV-1 HXB2 env 662 - 667 aa) and 4E10 (NWFDIT, HIV-1 HXB2 env 671 - 676 aa) in the membrane proximal-external region of gp41 from clade B' HIV-1-infected individuals living in Henan province, China. We also examined the frequency of a mutation and its relation to disease progression. METHODS: A cohort of 54 treatment-naïve HIV-1-infected individuals was recruited in this study, and 16 individuals were selected for a short-term longitudinal study on sequence evolution. The HIV-1 env gp41 gene was amplified, cloned, and sequenced, and predicted amino acid sequences were aligned for analysis. RESULTS: The mutations E662A and K665E on the 2F5 epitope and N671S and T676S on the 4E10 epitope were seen. Simultaneous RNA sequencing showed some discrepancies with proviral DNA sequences. In our longitudinal study, mutation levels of these two neutralizing epitopes were low but diverse and persistent. The frequencies of mutations within the 4E10 peptide NWFDIT in slow progressors were noticeably lower than those in AIDS patients (P < 0.05). CONCLUSIONS: Antigenic variation of the neutralizing epitopes 2F5 and 4E10 is limited in subtype B' infection, and that 4E10 peptide mutation is correlated with disease progression. Monitoring epitope mutations will offer useful data for development of the candidate 2F5-like and 4E10-like antibodies to prevent and treat AIDS.