Objective:To explore the clinical and genetic characteristics of two children with mental retardation and microcephaly.Methods:Two children who had visited the Anhui Children′s Hospital respectively on March 12 and June 22, 2021 were selected as the study subjects. Peripheral venous blood samples were collected from them and their parents, and subjected to chromosomal karyotyping and whole exome sequencing analyses. Candidate variants were verified by Sanger sequencing and pathogenicity analysis. This study was approved by the Anhui Children′s Hospital (Ethics No. EYLL-2018-008).Results:Chromosomal karyotyping and copy number detection of the two children had found no abnormality. Whole exome sequencing revealed that child 1 has harbored a c. 471delT (p.Pro157Profs*9) frameshifting variant of the CASK gene, whilst child 2 has harbored a c. 1259_1269delCTGAGAATAAC (p.Pro420fs*27) frameshifting variant of the CASK gene. Sanger sequencing confirmed that both variants were de novo in origin. Based on the guidelines from the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG-AMP), both variants were rated as pathogenic (PVS1+ PS2+ PP3). Conclusion:The de novo variants of the CASK gene probably underlay the pathogenesis of mental retardation and microcephaly in both children.

Background and objectiveWIF-1 is an important tumor-suppressing gene in lung cancer, and its encoding protein WIF-1 can reduce proliferation and promote apoptosis by inhibiting Wnt/β-catenin signaling in lung can-cer. hTis study constructs a eukaryotic expression plasmid carrying WIF-1 using FDA-approved clinical plasmid pVAX and explores the anti-tumor effect of pVAX-WIF-1 on A549 lung cancer cellsin vitro andvivo.MethodshTe DNA fragment of human WIF-1 coding sequence was ampliifed by PCR and was cloned into the multiple cloning sites of eukaryotic expression vector pVAX to construct pVAX-WIF-1. A recombinant plasmid was transfected into lung cancer A549 cells, and the expression ofWIF-1 genes was veriifed by Western blot atfer transfection. Subsequently, the effect of pVAX-WIF-1 on cell apoptosis and proliferation was identiifed by MTT assay, staining A549 cells with Hoechst 3235, and lfow cytometry. Finally, the A549 sub-cutaneous xenogratf was established to detect the effect of pVAX-WIF-1 on lung tumor growthin vivo.Results hTe results of restriction enzyme digestion, PCR, and sequencing indicated that eukaryotic expression plasmid pVAX-WIF-1 was successfully constructed. hTe protein expression level of WIF-1 was increased in the transfected A549 cells. Further results showed that transfection with pVAX-WIF-1 signiifcantly inhibited proliferation and promoted apoptosis in A549 cells. Moreover, pVAX-WIF-1 signiifcantly inhibited the tumor growth of the A549 subcutaneous xenogratfin vivo.ConclusionhTe recombinant eukaryotic expression vector pVAX-WIF-1 was successfully constructed. Transfection with pVAX-WIF-1 could signiifcantly inhibit proliferation and promote apoptosis of lung cancer A549 cells and also effectively inhibit the tumor growth of the A549 subcutaneous xenogratfin vivo. Our research can contribute to clinical applications of WIF-1 in lung cancer gene therapy.

Objective:To investigate the distributions of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) genotypes among newly reported HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province from 2016 to 2019.Methods:A total of 1 289 newly reported HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture were collected through the National Acquired Immunodeficiency Syndrome Comprehensive Prevention and Control Data Information System From January 2016 to December 2019. Among them, 996 subjects with a plasma volume of ≥200 μL were selected to perform HIV and HCV genotyping. The HIV pol gene, the HCV core protein-binding envelope protein ( CE1) gene and non-structural protein 5B ( NS5 B) gene were amplified using the nested polymerase chain reaction.The phylogenetic tree was constructed by MEGA 7.0 software to classify the genotypes. Chi-square test was used for statistical analysis. Trend chi-square test was used to analyze the trend of HIV and HCV genotypes. Results:Among the 996 cases with HIV/HCV co-infection, HIV and HCV sequences from a total of 554 subjects (55.6%, 554/996) were successfully obtained, and the genotypes of HIV and HCV were diverse. HIV genotype C (40.3%, 223/554) and BC recombinant (33.6%, 186/554) were the most prevalent, followed by genotype B (6.5%, 36/554) and circulating recombinant form (CRF)01_AE (3.6%, 20/554). HCV genotype 3b was the most prevalent (31.2%, 173/554), followed by genotype 6u (19.5%, 108/554), 1a (17.5%, 97/554), 6n (11.4%, 63/554), 3a (8.7%, 48/554) and 6xg (6.3%, 35/554). The prevalence of HIV genotype C showed a downward trend ( χtrend2=7.23, P<0.001), while the prevalence of BC recombinant showed an upward trend ( χtrend2=5.97, P<0.001), and the proportion of BC recombinant was higher than genotype C in 2019 (54.9%(101/184) vs 21.7%(40/184)). However, there were no statistically significant differences in the proportions of genotype 3b, 6u and 1a from 2016 to 2019 ( χtrend2=1.43, 1.79 and 0.39, respectively, P=0.152, 0.074 and 0.695, respectively). The HIV genotype distribution among patients with different ethnic groups were significantly different ( χ2=22.06, P=0.037). Conclusions:The diversity of HIV and HCV genotypes is high and complex among HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture. BC recombinant shows a trend of becoming the predominant HIV genotype among these co-infected patients. Therefore, surveillance of the prevalence of HCV and HIV genotypes in Burmese population needs to be further strengthened.

ObjectiveTo determine the trend and influencing factors of HIV subtypes in newly reported HIV/AIDS cases in Dehong Dai and Jingpo Autonomous Prefecture （Dehong Prefecture） from 2017 to 2019. MethodsRNA extraction was conducted among newly reported HIV/AIDS cases in Dehong Prefecture from 2017 to 2019 whose plasma volume was more than 200 μL. The gag， env and pol genes were amplified by using RT-PCR and then sequenced to determine the subtypes. ResultsA total of 3 287 HIV-infected cases were newly reported in Dehong from 2017 to 2019. The HIV gag， env and pol genotypes were determined in 1 813 cases. The major subtypes were subtype C （28.4%，515/1 813）， recombination form BC （22.0%，398/1 813） and CRF_01AE （18.1%，329/1 813）. Furthermore， the proportion of subtype B， subtype C and CRF01_AE decreased over years， whereas 01/BC， CRF07_BC and CRF08_BC increased over years in both Chinese and Burmese patients （χ²=75.212，P<0.001）. There were significant differences in gender， age， marital status， ethnicity， educational level and transmission route among Chinese and Burmese HIV-infected cases with diverse HIV genotypes （all P<0.05）. ConclusionHIV subtypes in Dehong change over time， which demonstrates that the proportion of BC recombinant subtypes and unique recombinant subtypes increased significantly.

ObjectiveThis study aimed to investigate the HIV genotypic subtypes and molecular transmission clusters among men who have sex with men （MSM） with newly reported HIV infections in Dehong Dai and Jingpo Autonomous Prefecture （Dehong Prefecture）， Yunnan Province， China， between 2010 and 2019. The study aimed to identify potential high-risk transmitters and provide reference data for screening， management， and intervention of infection sources. MethodsPlasma samples from newly reported HIV-positive MSM individuals in Dehong Prefecture between 2010 and 2019 were collected. The viral pol gene fragments were amplified， sequenced， and genotyped. Genetic distances （GD） between pairwise sequences were analyzed and calculated. MEGA 7.0 and Gephi were used for phylogenetic and molecular transmission network analysis. ResultsA total of 159 newly reported HIV infections among MSM were included in the study， with successful genotyping of 100 cases. Nine HIV-1 subtypes were identified， with the most prevalent being CRF01_AE subtype （52%）， followed by CRF07_BC subtype （31%）， CRF55_01B subtype （10%）， and others （7%）. Cluster analysis revealed a total network access rate of 67%， forming three transmission clusters. CRF01_AE subtype formed two transmission clusters with 38 and 3 infected individuals， while CRF07_BC subtypes formed one transmission cluster with 26 infected individuals. The transmission network within the CRF01_AE clusters exhibited a more complex relationship. Significant differences in educational level were observed between the two main transmission clusters. ConclusionThe predominant HIV subtypes among newly reported MSM cases in Dehong Prefecture between 2010 and 2019 were CRF01_AE and CRF07_BC. Significant cultural differences are observed between the main transmission clusters. Continued monitoring of genotypic subtypes and targeted interventions within transmission clusters are warranted.