A novel internet sampling for HIV surveillance: feasibility of self-sampling and preparation of DBS for delivery detection of HIV total nucleic acid and complementarity to sentinel surveillance.

BACKGROUND: To propose a new mode of HIV test and surveillance among population of men who have sex with men (MSM): Internet-based Self-sampling at home plus Laboratory testing of HIV total nucleic acid (TNA) in dried blood spot (DBS) (ISL of DBS TNA). Feasibility of ISL of DBS TNA was studied. Characteristics of the new mode and that of conventional surveillance mode at HIV voluntary counseling and testing clinic (VCT) were compared. METHODS: A non-governmental organization (NGO) published the recruitment information on the WeChat public account. MSM filled in the questionnaire online, applied for self-sampling service package, and mailed the self made DBS to professional laboratory. The laboratory performed HIV TNA test and submitted the test results to online platform. Participants queried test results online with their unique ID. Center for Disease Control and Prevention (CDC) followed up participants with positive nucleic acid results using IDs and contact information. Rates were compared by using the Chi-Square test or Fisher's exact test. RESULTS: Four hundred twenty-three questionnaires were completed. 423 self-sampling service packages were sent out and 340 DBSs were returned to professional laboratory within one month with qualified rate of sampling as high as 95.0% (323/340). Seven samples were found to be TNA positive. Comparing ISL of DBS TNA with sentinel surveillance, it was found that there was a significant difference in the composition ratio of the two modes of surveillance population (P < 0.05). HIV prevalence of ISL of DBS as 2.17% was significantly lower than sentinel site as 8.96% (χ2 = 14.953, P = 0.000 < 0.05). CONCLUSIONS: ISL of DBS TNA proposed is feasible and has a high self-sampling qualification rate, good confidentiality. It is an effective supplement to routine sentinel surveillance and has important promotion value.

Biological Characteristics and Pathogenicity Analysis of a Low Virulence G2a Porcine Epidemic Diarrhea Virus.

Since the outbreak caused by a porcine epidemic diarrhea virus (PEDV) variant in 2010, the current epidemic of PEDV genotype 2 (G2) has caused huge economic losses to the pig industry in China. In order to better understand the biological characteristics and pathogenicity of the current PEDV field strains, 12 PEDV isolates were collected and plaque purified during 2017 to 2018 in Guangxi, China. The neutralizing epitopes of the spike proteins and the ORF3 proteins were analyzed to evaluate genetic variations, and they were compared with the reported G2a and G2b strains. Phylogenetic analysis of the S protein showed that the 12 isolates were clustered into the G2 subgroup (with 5 and 7 strains in G2a and G2b, respectively) and that they shared 97.4 to 99.9% amino acid identities. Among them, one of the G2a strains, CH/GXNN-1/2018, which had a titer of 106.15 PFU/mL, was selected for pathogenicity analysis. Although piglets infected with the CH/GXNN-1/2018 strain exhibited severe clinical signs and the highest level of virus shedding within 24 h postinfection (hpi), recovery and decreased virus shedding were seen after 48 hpi, and no piglets died during the whole process. Thus, the CH/GXNN-1/2018 strain had low virulence in suckling piglets. Virus neutralizing antibody analysis showed that the CH/GXNN-1/2018 strain induced cross-protection against both homologous G2a and heterologous G2b PEDV strains as early as 72 hpi. These results are of great significance for understanding PEDV in Guangxi, China, and they provide a promising naturally occurring low-virulent vaccine candidate for further study. IMPORTANCE The current epidemic of porcine epidemic diarrhea virus (PEDV) G2 has caused huge economic losses to the pig industry. Evaluation for low virulence of the PEDV strains of subgroup G2a would be useful for the future development of effective vaccines. In this study, 12 field strains of PEDV were obtained successfully, and they were characterized from Guangxi, China. The neutralizing epitopes of the spike proteins and the ORF3 proteins were analyzed to evaluate antigenic variations. One of the G2a strains, CH/GXNN-1/2018, was selected for pathogenicity analysis, and it showed that the CH/GXNN-1/2018 strain had low virulence in suckling piglets. These results provide a promising naturally occurring low-virulent vaccine candidate for further study.

Phylogenetic and Spatiotemporal Analyses of Porcine Epidemic Diarrhea Virus in Guangxi, China during 2017-2022.

Since 2010, porcine epidemic diarrhea virus (PEDV) has swept across China and spread throughout the country, causing huge economic losses. In this study, 673 diarrhea samples from 143 pig farms in Guangxi during 2017-2022 were collected and detected for PEDV. Ninety-eight strains were selected for S1 gene analyses and these strains were classified into four subgroups (G1b, G2a, G2b and G2c), accounting for 1.02 (1/98), 75.51 (74/98), 16.33 (16/98) and 7.14% (7/98) of the total, respectively. Importantly, an increased number of strains in the G2c subgroup was found from 2019 onwards. Bayesian analysis revealed that Guigang may have been the epicenter of PEDVs in Guangxi. In addition, Guigang was identified as the primary hub from which PEDVs spread via two routes, namely Guigang-Wuzhou and Guigang-Laibin. Moreover, several coinfections of novel PEDV variants bearing large deletions in the partial S1 protein and PEDVs possessing an intact partial S1 protein were found in pigs. Further recombination analyses indicated that two of the strains, 18-GXNN-6 and 19-GXBH-2, originated from intra-genogroup recombination. Together, our data revealed a new profile of PEDV in Guangxi, China, which enhances our understanding of the distribution, genetic characteristics and evolutionary profile of the circulating PEDV strains in China.

Preparation of beta-cyclodextrin based nanocomposite for magnetic solid-phase extraction of organic ultraviolet filters.

A simple and efficient analytical method for organic UV filters (UV-Fs) in environmental samples has been established in this study. Taking advantage of the hydrophobicity on the inner cavity, hydrophilicity on the outer wall, and host-guest interaction provided by beta-cyclodextrin, a core-shell magnetic extraction material was firstly synthesized by using a facile method. The extractant was utilized in magnetic solid-phase extraction of UV-Fs in complex environmental samples, including beach sand, sediment and river water samples, followed by the quantitation using high-performance liquid chromatography. A series of factors affecting extraction efficiencies of seven UV-Fs were profoundly optimized. Under the optimal conditions, the linear ranges were at 5.0-5.0 × 102 ng mL-1 for the UV-Fs with regression coefficients (r) at 0.9984-0.9998. The limits of detection were from 0.12 to 1.4 ng mL-1. The recoveries were in the range of 84.2-109%. Furthermore, the molecular dynamics simulations and independent gradient model analysis were applied to reveal the adsorption configuration and interaction mechanisms between target analytes and the sorbent.

Comparative Characterization and Pathogenicity of a Novel Porcine Epidemic Diarrhea Virus (PEDV) with a Naturally Occurring Truncated ORF3 Gene Coinfected with PEDVs Possessing an Intact ORF3 Gene in Piglets.

Coinfection caused by various genotypes of porcine epidemic diarrhea virus (PEDV) is a new disease situation. We previously reported the coexistence of PEDV strains containing different ORF3 genotypes in China. In this study, the PEDV strains 17GXCZ-1ORF3d and 17GXCZ-1ORF3c were isolated and plaque-purified from the same piglet, which had a natural large deletion at the 172-554 bp position of the ORF3 gene or possessed a complete ORF3 gene, respectively. Meanwhile, 17GXCZ-1ORF3d had >99% nt identity with 17GXCZ-1ORF3c in the 5'UTR, ORF1a/1b, S, E, M, N and 3'UTR regions but only demonstrated low nucleotide identities (80.5%) in the ORF3 gene. To elucidate the pathogenicity, 7-day-old piglets were infected. Piglets infected with these two PEDV strains exhibited severe clinical signs and shed the virus at the highest level within 96 hpi. Compared with the piglets inoculated with the 17GXCZ-1ORF3c strain, the piglets inoculated with the 17GXCZ-1ORF3d strain had higher mortality rates (75% vs. 50%), an earlier onset of clinical signs with a significantly higher diarrhea score, lower VH:CD ratios and a higher percentage of PEDV-positive enterocytes. This study is the first to report PEDV coinfections with different ORF3 genotypes, and a PEDV strain with a large deletion in the ORF3 gene might have the advantage of a potential genetic marker, which would be useful during vaccine development.

Genetic Diversity of Porcine Epidemic Diarrhea Virus With a Naturally Occurring Truncated ORF3 Gene Found in Guangxi, China.

Porcine epidemic diarrhea virus (PEDV) is one of the major enteric pathogens, causing severe enteric disease, resulting in enormous economic losses. The ORF3 gene encodes an accessory protein which is related to the infectivity and virulence of PEDV. In this study, 33 PEDV positive field samples were collected from Guangxi, from 2017 to 2019, and the genetic diversity of ORF3 was investigated. Thirty-eight strains of ORF3 were obtained, and these were composed of five strains of ORF3 named Guangxi naturally truncated strains that were 293 bp in length, with continuous deletions from 172 to 554 bp. The Guangxi naturally truncated strains encoded a truncated protein of 89 amino acids, which had clustered into a new group referred to as Group 3, and these might be involved in the variations of virulence. Three genotypes (G1-1 subgroup, G1-3 subgroup, and Group 3) existed simultaneously in Guangxi based on the genetic and evolutionary analysis of the ORF3 gene. The sequence information in the current study will hopefully facilitate the establishment of a diagnostic method that can differentiate the PEDV field stains. Continued surveillance will be useful for monitoring PEDV transmission. Differentiation of the ORF3 genes in PEDV field strains can help us to choose an appropriate PEDV vaccine candidate in the future and prevent outbreaks of PED more effectively.

Trends of HIV incidence and prevalence among men who have sex with men in Beijing, China: Nine consecutive cross-sectional surveys, 2008-2016.

BACKGROUND: Sexual transmission of HIV among men who have sex with men (MSM) increased markedly in China during the past decade. HIV incidence is a critical indicator in HIV surveillance and we use a HIV-1 BED-capture-enzyme immunoassay (BED-CEIA) to examine the incidence among MSM in Beijing from 2008 to 2016. Risk factors related to recent HIV infection were also assessed. METHODS: Consecutive cross-sectional surveys on MSM were conducted yearly from 2008 through 2016. Demographic and behaviors data were collected. HIV status was determined and HIV positive specimens were tested for recent infection using BED-CEIA. Specimens with ODn values≤0.8 were considered recently infected, HIV incidence rates and prevalence were then calculated. Risk factors associated with recent HIV infection were assessed by univariate and multivariable logistic regression. RESULTS: From 2008 to 2016, the numbers of eligible participants in the nine consecutive years ranged from 472 to 616. All the 261 eligible HIV-positive specimens were subjected to recent HIV infection testing. HIV prevalence ranged from 5.0% (3.3%-6.8%) to 10.2% (7.8%-12.7%), and incidence ranged from 1.57% (0.19%-2.95%) to 6.63% (3.65%-9.61%). MSM who never or sometimes used condoms during anal sex with men in the past 6 months (aOR = 1.515, 95%CI: 1.016-2.257, p = 0.041), or having syphilis infection (aOR = 1.561, 95%CI: 0.946-2.575, p = 0.081) were more likely to be recently infected with HIV. Being a Beijing resident (aOR = 0.409, 95%CI: 0.212-0.790, p = 0.008), or having only one male anal sex partner in the past 6 months (aOR = 0.467, 95%CI: 0.220-0.994, p = 0.048) were associated with a lower risk for recent HIV infection. CONCLUSIONS: The HIV incidence fluctuated among MSM in Beijing. Unprotected anal sex, having multiple sex partners, being a non-registered Beijing resident and having a syphilis infection play important roles in the recent HIV infection. Effective intervention measures for HIV and syphilis control and prevention should be continuously strengthened.

Human fetal intestinal epithelial cells metabolize and incorporate branched chain fatty acids in a structure specific manner.

BACKGROUND: Branched chain fatty acids (BCFA) are constituents of gastrointestinal (GI) tract in healthy newborn human infants, reduce the incidence of necrotizing enterocolitis (NEC) in a neonatal rat model, and are incorporated into small intestine cellular lipids in vivo. We hypothesize that BCFA are taken up, metabolized and incorporated into human fetal cells in vitro. METHODS: Human H4 cells, a fetal non-transformed primary small intestine cell line, were incubated with albumin-bound non-esterified anteiso-17:0, iso-16:0, iso-18:0 and/or iso-20:0, and FA profiles in lipid fractions were analyzed. RESULTS: All BCFA were readily incorporated as major constituents of cellular lipids. Anteiso-17:0 was preferentially taken up, and was most effective among BCFA tested in displacing normal (n-) FA. The iso BCFA were preferred in reverse order of chain length, with iso-20:0 appearing at lowest level. BCFA incorporation in phospholipids (PL) followed the same order of preference, accumulating 42% of FA as BCFA with no overt morphological signs of cell death. Though cholesterol esters (CE) are at low cellular concentration among lipid classes examined, CE had the greatest affinity for BCFA, accumulating 65% of FA as BCFA. BCFA most effectively displaced lower saturated FA. Iso-16:0, iso-18:0 and anteiso-17:0 were both elongated and chain shortened by ±C2. Iso-20:0 was chain shortened to iso-18:0 and iso-16:0 but not elongated. CONCLUSIONS: Nontransformed human fetal intestinal epithelial cells incorporate high levels of BCFA when they are available and metabolize them in a structure specific manner. These findings imply that specific pathways for handling BCFA are present in the lumen-facing cells of the human fetal GI tract that is exposed to vernix-derived BCFA in late gestation.

[Analysis on factors associated with taking subsequent confirmation test among men who have sex with men after being tested positive in oral fluid HIV antibody test in Beijing].

OBJECTIVE: To analyze the proportion and associated factors of taking subsequent confirmation test among men who have sex with men (MSM) after being tested positive in oral fluid HIV antibody test. METHODS: By using successive sampling, 1 003 MSM, who were tested positive in oral fluid HIV antibody test in China-Bill & Melinda Gates Foundation AIDS prevention Program (Extension program) in Beijing during May 1 to December 31, 2013, were recruited. The inclusion criteria included: the objects were men who reported having sex with men; the objects aged more than 18 years old; the objects were tested positive in oral fluid HIV antibody test; the objects had not been reported as HIV positives in China Information System for Disease Control and Prevention previously. According to the program strategy, MSM grassroots organizations transferred the respondents to seek subsequent confirmation tests in specific Center for Disease Control and Prevention (CDCs) or hospitals. The subsequent confirmation tests included: fingertip blood HIV antibody rapid test, venous blood Enzyme Linked Immunosorbent Assay (ELISA) HIV antibody test and venous blood Western Blot (WB) HIV antibody test. Chi-square test was adopted to compare the proportion of taking subsequent confirmation tests in different groups. Nonconditional multivaritae binarylogistic regression analysis was taken to identify the associated factors with whether taking subsequent confirmation tests and to calculate the OR (95% CI) values. RESULTS: The 1 003 respondents were (30.9 ± 9.1) years old. Among all objects, 87.8% (881/1 003) of them took fingertip blood HIV antibody rapid tests and the positive rate was 85.4% (752/881). 98.0% (737/752) of those who were identified as positive in fingertip blood HIV rapid tests took ELISA and WB tests, and the positive rate was 94.4% (696/737). Comparing with those who were expected to seek subsequent confirmation tests in CDCs, the OR (95% CI) value of those who were expected to seek tests in hospitals was 5.10 (1.69-15.36). The OR (95% CI) values of those who used condom sometimes and those who never used condom in anal sex were 5.81 (2.14-15.77) and 3.45 (2.00-5.97) respectively, in comparison with those who reported not having anal sex or using condom consistently in anal sex during the past 6 months. Comparing with the respondents recruited from the internet, the OR (95% CI) values of those recruited in bathrooms, parks/toilets and bars were 0.17 (0.05-0.53), 0.10 (0.04-0.29) and 0.22 (0.06-0.79) respectively. The likelihood of taking subsequent confirmation test decreased with the increase of number of male sexual partners in the past 3 months, and the OR (95% CI) value was 0.92 (0.86-0.99). CONCLUSION: The potential HIV positive MSM in the bathroom, park/toilet and bars are less likely to take subsequent confirmation test. Those who do not use condom consistently during anal sex are more likely to seek subsequent confirmation test. Medical organization conducting subsequent confirmation tests is more likely to increase the confirmation test rate of potential HIV positive MSM. The number of male sexual partners has negative correlation with whether to accept the subsequent confirmation test.

Phylogenetic and temporal dynamics of human immunodeficiency virus type 1B in China: four types of B strains circulate in China.

To investigate the origin and evolutionary history of the spread of HIV-1 subtype B in China, a total of 409 sequences of pol gene sampled from 1994 to 2012 in 29 provinces across China was subjected to phylogenetic and Bayesian molecular clock analyses. The study reveals that subtype B strains in China are genetically diverse and can be classified into four distinct subgroups, namely B' (Thai-B), BJ-B (Beijing-B), Pan-B (Pandemic-B), and TW-B (Taiwan-B), according to the origin of the sequences. The BJ-B and TW-B are reported for the first time. Phylogeographic analysis reveals that B' exhibits a nationwide, transprovincial distribution, and is found in 21 provinces in China in this study, whereas the Pan-B, BJ-B, and TW-B lineages are restricted to particular regions. From the same common ancestor of B', there arise two subclusters in which sequences from Yunnan occupy the basal position. The times of the most recent common ancestors (tMRCAs) of B' and BJ-B are estimated to be 1983.6 (1975.9-1990.3) and 1995.3 (1989.6-2000.3), respectively. The skyline plot profile reveals an exponential decrease in median number of effective infections of subtype B in China from 1994 to 2009. The existence of four types of B clades also indicates distinct transmission networks of subtype B, originating from different introduction events at different time points. The data presented here offer a new perspective on the epidemic of HIV-1 subtype B in China.

Promising antibody testing strategies for early infant HIV infection diagnosis in China.

BACKGROUND: In China, 1.1% of people living with HIV were transmitted vertically, causing a heavy burden on families and society. Early infant diagnosis (EID) is critical for improving neonatal survival. The purpose of this study is to suggest improvement in antibody testing strategies with dried blood spots (DBSs) for EID in China through analysis of anti-HIV seroreversion of infants. METHODS: A total of 280 infants born to HIV infected mothers in four diverse provinces of China where multiple subtypes coexist were enrolled. The status of the infants' infection was determined by HIV antibody enzyme immunoassay and Western blot analysis at ≥ 18 months of age or by convincing clinical and epidemiologic data for deceased infants. A total of 1028 DBSs were collected during follow-up, which were tested to obtain anti-HIV signal to cut-off ratio (S/CO) data. RESULTS: For uninfected infants, anti-HIV S/CO decreased with age. Seropositivity percentage declined most rapidly at 6 months to 9 months of age and 98.7% children seroreverted by 12 months of age. For most infected infants, minimum S/CO values were obtained at ≤ 6 months of age. Antibody negative predictive value was 100% at ≥ 6 months of age. An S/CO increase ≥ 1.86 after three months follow-up can determine HIV infection. S/CO threshold of 3.17 can differentiate infected from uninfected infants for exposed kids at 9 months or older with sensitivity as 100% and specificity ≥ 94.2%. SIGNIFICANCE: Suggestions obtained through studying seroreversion data of Chinese HIV-exposed infants help improve antibody strategies for HIV EID in China. The infection can be determined as early as 3 months of age and excluded as early as 6 months of age.

[Molecular epidemiological characteristics of HIV-1 strains isolated from newly diagnosed MSM subjects (2006-2010) in Beijing, China].

This study aims to analyze the molecular epidemiological characteristics of HIV-1 strains prevailing among men who have sex with men (MSM) in Beijing, China. The pol gene fragments from 250 newly diagnosed HIV-1-infected MSM individuals during 2006-2010 in Beijing were amplified by RT-nested PCR, sequenced, and phylogenetically analyzed. HIV-1 pol gene from 189 individuals were amplified and analyzed; 81 (42. 9%), 3 (1. 6%), 2 (1.0%), 88 (46. 6%), and 15 (7.9%) individuals were infected with HIV-1 subtypes B, B', C, CRF01_AE, and CRF07_BC, respectively. The subtypes B and CRF01_AE could both be grouped into two clusters, and CRFO7_BC strains shared high homology and were presumed to originate from a common ancestor. The HIV-1 circulating in MSM in Beijing had a lower genetic diversity than in heterosexuals. The HIV-1 epidemic (2006-2010) in MSM in Beijing was actually a rapid spread of HIV-1 CRF01 AE and B, or rather native strains of the two viruses.

[Epidemiological analysis on recent infected HIV-1 patients among newly reported HIV cases in Beijing, from 2009 to 2011].

OBJECTIVE: To understand the epidemiologic characteristics on newly reported HIV cases and those recently infected HIV-1 cases in the past few years in Beijing so as to find out their correlates. METHODS: All the qualified newly reported HIV serum samples from both the first quarters of 2009, 2010 and the second quarter of 2011 in Beijing, were tested using the BED HIV-1 incidence capture enzyme immunoassay (BED-CEIA) to identify those recent HIV-1 infection (BED positive) samples and related socio-demographic characteristics. The proportions of BED positives were determined and the correlates analyzed by SPSS software. RESULTS: The Annual qualified newly reported HIV serum samples of each year from Beijing were 274, 236 and 356, with BED positive rate among them as 35.0%, 31.8% and 33.7%, respectively. The majority of newly reported HIV cases of each year were males (89.1%, 88.6%, 91.9%), range of age as 20-39 years (77.4%, 75.8%, 82.0%), Han nationality (77.7%, 88.6%, 85.7%), and most of them were not permanent residents of Beijing (74.1%, 80.5%, 2011 data missed). The proportions of patients through sexual transmission showed upward trend (67.5%, 76.2%, 86.8%). Especially in the 'man having sex with man' (MSM)group, it showed a significantly increase (44.9% , 45.3% , 62.6%). The proportions of injection drug users (15.3%, 8.1%, 9.0%)declined to some extent during this period. Data from statistics indicated that the proportions of BED positives among newly reported HIV cases of each year were significantly correlated to factors as patients' sex, transmission route and source of samples, but not significantly correlated to patents' age or marital status. Male cases accounted for 96.9% and MSM accounted for 73.5% of all the BED-positives during 2009-2011. CONCLUSION: MSM accounted for both large portion of newly reported HIV cases and recent HIV-1 infections which suggested an increasing trend in 2009-2011. It seemed that the characteristic and changing trends of HIV epidemic in Beijing was oriented by MSM population in the last years.

[Molecular epidemiological characteristics of HIV-1 B'/C strains in Beijing].

OBJECTIVE: To analyze the molecular epidemiological characteristic of HIV-1 B'/C strains prevalent in Beijing. METHODS: Plasma samples were collected from 200 newly diagnosed HIV-1 B'/C individuals reported during 2006 to 2010 in Beijing. The gag gene fragments were amplified from RNA template extracted from plasma with reverse transcription (RT) and nested polymerase chain reaction (PCR). And the sequences were analyzed by phylogenetic methods and Entropy analysis. RESULTS: A total of 159 sequences were successfully amplified from the gag genes of which 147 was CRF07_BC and 12 CRF08_BC. There were 3 main sub-clusters in CRF07_BC phylogenetic tree and they were named as sub-cluster IDU-Max (89 sequences), sub-cluster IDU-Min (22 sequences) and sub-cluster MSM (34 sequences) based on transmission.No international reference strain was closely related with these three sub-clusters except for one strain identified in Taiwan. All CRF07_BC recombinant strains were remarkable for their low interpatient diversity in gag genes (3.7%, 3.3% and 2.0% for isolates from IDU-Max, IDU-Min and MSM respectively).When compared with sub-cluster IDU-Max, there were 32 and 41 significantly different sites of nucleotide polymorphism compositions in sub-clusters IDU-Min and MSM. CONCLUSION: This is the first report of describing the existence of three main epidemic sub-clusters in CRF07_BC strains prevalent in Beijing. And IDU-Max sub-cluster is the dominant strain. The CRF07_BC in Beijing are less diverse than other strains and may be derived from a common ancestor.

[Molecular-epidemiological characteristics of HIV-1 isolated from newly diagnosed female subjects in Beijing, 2006 - 2010].

OBJECTIVE: To analyze the molecular-epidemiological characteristics of HIV-1 strains prevailing among female people living with HIV in Beijing. METHODS: Gag gene fragments from the 100 newly diagnosed female HIV-1 infections during 2006 to 2010 in Beijing were amplified, sequenced, and phylogenetically analyzed. RESULTS: Eighty-two HIV-1 gag gene fragments were amplified and analyzed. 1 (1.22%), 1 (1.22%), 3 (3.66%), 23 (28.05%), 8 (9.76%), 2 (2.44%), 1 (1.22%), 18 (21.95%), 3 (3.66%), 1 (1.22%), 14 (17.07%), 4 (4.88%) and 3 (3.66%) individuals were infected with HIV-1 subtypes A1, A2, B, B', C, D, G, H, CRF01_AE, CRF02_AG, CRF07_BC, CRF08_BC and B'/C recombinants respectively. CONCLUSION: The subtypes circulating in female HIV infections in Beijing were more diverse than in male and the proportions of B' and rare subtypes were relatively high. Surveillance programs on HIV-1 genetic diversity should be strengthened.

Phylogenetic and temporal dynamics of human immunodeficiency virus type 1 CRF01_AE in China.

To explore the epidemic history of HIV-1 CRF01_AE in China, 408 fragments of gag gene sequences of CRF01_AE sampled in 2002-2010 were determined from different geographical regions and risk populations in China. Phylogenetic analysis indicates that the CRF01_AE sequences can be grouped into four clusters, suggesting that at least four genetically independent CRF01_AE descendants are circulating in China, of which two were closely related to the isolates from Thailand and Vietnam. Cluster 1 has the most extensive distribution in China. In North China, cluster 1 and cluster 4 were mainly transmitted through homosexuality.The real substance of the recent HIV-1 epidemic in men who have sex with men(MSM) of North China is a rapid spread of CRF01_AE, or rather two distinctive natives CRF01_AE.The time of the most recent common ancestor (tMRCA) of four CRF01_AE clusters ranged from the years 1990.9 to 2003.8 in different regions of China. This is the first phylogenetic and temporal dynamics study of HIV-1 CRF01_AE in China.

Quality assurance in the HIV/AIDS laboratory network of China.

BACKGROUND: In 2009, there were 8273 local screening laboratories, 254 confirmatory laboratories, 35 provincial confirmatory central laboratories and 1 National AIDS Reference Laboratory (NARL) in China. These laboratories were located in Center for Disease Control and Prevention (CDC) facilities, hospitals, blood donation clinics, maternal and child health (MCH) hospitals and border health quarantine health-care facilities. METHODS: The NARL and provincial laboratories provide quality assurance through technical, bio-safety and managerial training; periodic proficiency testing; on-site supervisory inspections; and commercial serologic kit evaluations. RESULTS: From 2002 to 2009, more than 220 million HIV antibody tests were performed at screening laboratories, and all reactive and indeterminate samples were confirmed at confirmatory laboratories. The use of highly technically complex tests, including CD4 cell enumeration, viral load, dried blood spot (DBS)-based early infant diagnosis (EID), drug resistance (DR) genotyping, HIV-1 subtyping and incidence assays, have increased in recent years and their performance quality is closely monitored. CONCLUSION: China has made significant progress in establishing a well-coordinated HIV laboratory network and QA systems. However, the coverage and intensity of HIV testing and quality assurance programmes need to be strengthened so as to ensure that more infected persons are diagnosed and that they receive timely prevention and treatment services.

Analysis of the statistical thermodynamic model for nonlinear binary protein adsorption equilibria.

The statistical thermodynamic (ST) model was used to study nonlinear binary protein adsorption equilibria on an anion exchanger. Single-component and binary protein adsorption isotherms of bovine hemoglobin (Hb) and bovine serum albumin (BSA) on DEAE Spherodex M were determined by batch adsorption experiments in 10 mM Tris-HCl buffer containing a specific NaCl concentration (0.05, 0.10, and 0.15 M) at pH 7.40. The ST model was found to depict the effect of ionic strength on the single-component equilibria well, with model parameters depending on ionic strength. Moreover, the ST model gave acceptable fitting to the binary adsorption data with the fitted single-component model parameters, leading to the estimation of the binary ST model parameter. The effects of ionic strength on the model parameters are reasonably interpreted by the electrostatic and thermodynamic theories. The effective charge of protein in adsorption phase can be separately calculated from the two categories of the model parameters, and the values obtained from the two methods are consistent. The results demonstrate the utility of the ST model for describing nonlinear binary protein adsorption equilibria.

Ionically crosslinked alginate/carboxymethyl chitin beads for oral delivery of protein drugs.

Complex beads composed of alginate and carboxymethyl chitin (CMCT) were prepared by dropping aqueous alginate-CMCT into an iron(III) solution. The structure and morphology of the beads were characterized by IR spectroscopy and scanning electron microscopy (SEM). IR confirmed electrostatic interactions between iron(III) and the carboxyl groups of alginate as well as CMCT, and the binding model was suggested as a three-dimensional structure. SEM revealed that CMCT had a porous morphology while alginate and their complex beads had a core-layer structure. The swelling behavior, encapsulation efficiency, and release behavior of bovine serum albumin (BSA) from the beads at different pHs were investigated. The BSA encapsulation efficiency was fairly high (>90%). It was found that CMCT disintegrated at pH 1.2 and alginate eroded at pH 7.4 while the complex beads could effectively retain BSA in acid (>85%) and reduce the BSA release at pH 7.4. The results suggested that the iron(III)-alginate-CMCT bead could be a suitable polymeric carrier for site-specific protein drug delivery in the intestine.

Quantitative determination of compounds in tobacco essential oils by comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry.

A quantitative analysis of the individual compounds in tobacco essential oils is performed by comprehensive two-dimensional gas chromatography (GC x GC) combined with flame ionization detector (FID). A time-of-flight mass spectrometer (TOF/MS) was coupled to GC x GC for the identification of the resolved peaks. The response of a flame ionization detector to different compound classes was calibrated using multiple internal standards. In total, 172 compounds were identified with good match and 61 compounds with high probability value were reliably quantified. For comparative purposes, the essential oil sample was also quantified by one-dimensional gas chromatography-mass spectrometry (GC/MS) with multiple internal standards method. The results showed that there was close agreement between the two analysis methods when the peak purity and match quality in one-dimensional GC/MS are high enough.