Construction of SARS-CoV-2 receptor binding domain molecular probe and its application in the isolation of neutralizing antibodies / 中国热带医学

@#Abstract: Objective　To construct SARS-CoV-2 receptor binding domain molecular probe for monoclonal memory B cell sorting and obtain RBD specific neutralizing antibodies from peripheral blood mononuclear cells (PBMCs) of COVID-19 convalescents by single-cell sorting. Methods　The SARS-CoV-2 RBD sequence was downloaded from GenBank, and the Avi tag and 6-histidine tags were added at the C-terminal. After codon optimization, it was chemically synthesized, cloned into the pDRVI1.0 vector, expressed after transfection of 293F cells, and biotinylated consequently. RBD-specific B cells were sorted out with this probe1 from the PBMCs of convalescents recovered from COVID-19. After B cells were lysed, the variable regions of heavy chain and light chain were amplified, cloned into the antibody expression vector, and transfected into 293F cells to express the antibody. Then the antibody was purified from the supernatant using protein A column and SARS-CoV-2 pseudovirus was used to test their neutralizing activity. Results　RBD-Avi probe was produced and successfully biotinylated sequentially with an efficiency of 30%-50%. Western blot analysis revealed that the biotinylated probe was recognized by the antibodies purified from COVID-19 convalescent plasma. Using this probe, 7 and 16 RBD-specific memory B cells were successfully isolated from the PBMCs of two convalescent individuals, accounting for 0.24% and 0.17% of the total cell population, respectively. After amplifying the variable regions of antibody heavy and light chains from the lysed B cells, 7 and 12 pairs of antibody heavy-light chains were obtained. A total of 16 antibodies were expressed in the convalescent individuals, and most of the purified antibodies showed neutralizing activity against the pseudovirus, with IC50 values of 6 antibodies below 1 μg/mL. The IC50 values of XJ-A9 and SCF-F1 against the wild-type pseudovirus were 0.07 μg/mL and 0.35 μg/mL, respectively. Conclusion　The SARS-CoV-2 RBD molecular probe constructed in this study has good antigenicity, and the isolated antibodies present neutralizing activity against wild-type SARS-CoV-2 pseudovirus.

Current status and considerations of sexuality education in primary and secondary school / 中国学校卫生

Abstract@#Comprehensive sexuality education is an important part of quality education, primary and secondary schools are the most suitable places for sex education. This paper sorts out the current status of sexuality education for primary and secondary school students in developed countries after presenting the overall significance of school based sexuality education, and further points out the problems and urgency of sexuality education for primary and secondary school students in China. It also put forward the way to new directions for advocacy, including the comphrehensive sexuality education curriculum system, training of sexuality education teachers, the positive and active role of families, as well as social and community support for sexuality education in schools.

Tuberculosis co-morbidity and perceptions about health care among HIV-infected plasma donors in rural China.

Limited community-based data exist about pulmonary tuberculosis (TB) comorbidity among HIV-infected individuals in China and no data exists about the TB burden in key high risk groups. We recruited 195 known HIV-infected plasma donors in one central China county and identified 9 (4.6%) active TB cases based on clinical assessment, including chest radiography. The low percentage of TB may be explained by improved immunity due to antiretroviral therapy.

Expanding research capacity and accelerating AIDS vaccine development in Asia.

According to the Joint UN Program on AIDS (UNAIDS), an estimated 4.9 million adults and children are living with HIV in Asia and the Pacific. Refinement and development of existing and new prevention and treatment technologies--including safe, effective, and accessible AIDS vaccines--are urgent public health priorities. The Asian region faces several challenges for AIDS vaccine development. There are multiple genetic variants of HIV-1 driving the epidemic in the region and too few vaccine candidates in the pipeline targeting those subtypes. Low HIV incidence throughout the region means that trial sites must recruit larger numbers of volunteers and shift their focus to higher-risk populations where incidence is higher. Also, the cultural, economic, and political diversity of the region may render collaboration very complex, but also beneficial at a regional level. Recognizing that collaborating as a region could foster and accelerate AIDS vaccine development, participants at the Sapporo International Consultation recommended that an AIDS Vaccine Asian Network (AVAN) be created to facilitate interactions between donors and funding opportunities, increase regional clinical trial and production capacity, support region-specific advocacy and communication strategies, contribute to the Global HIV Vaccine Enterprise Scientific Plan, prepare a regional approach for future vaccine deployment, and develop a regional platform for clinical trials including harmonized legal, regulatory, and ethical frameworks.

Predictors of unprotected sex among men who have sex with men in Beijing, China.

To estimate the prevalence of HIV and syphilis and to assess the predictors of unprotected anal intercourse (UAI) among men who have sex with men (MSM) in Beijing, a community-based survey recruited MSM in 2005 through internet advertisement, community outreach, and peer referral. Demographic, sexual, and HIV risk behavioral information were collected. Serospecimens were tested for HIV and syphilis infections. Of the 526 participants, 3.2% were HIV-positive, 11.2% syphilis-positive, 50% and 43.3% had UAI with regular and casual sex partners, respectively. Participants practicing UAI with regular male partners were independently associated with lower monthly income (adjusted odds ratio-AOR, 1.7; 95% CI, 1.0-3.0) and encountering male sex partners at bathhouses, public washrooms, and parks (AOR, 2.2; 95% CI, 1.0-4.9). Participants practicing UAI with casual male partners were associated with encountering male sex partners at bathhouses, publics washrooms, and park (AOR, 3.0; 95% CI, 1.8-5.2) and more male sex partners having receptive anal intercourse (AOR, 1.8; 95% CI, 1.1-2.9), and was inversely associated with receiving money for sex with men (AOR, 0.3; 95% CI, 0.2-0.7). Professional male sex workers were less likely to practice UAI in Beijing, suggesting the benefits of educational outreach to date. Further education, condom promotion, and prevention of sexually transmitted infections should be intensified urgently to combat the rising HIV epidemic among MSM in Beijing.