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BACKGROUND: Large-scale HIV genotype drug resistance study has not been conducted in Chongqing. METHODS: A retrospective study was conducted on people living with HIV(PLWH) who received HIV-1 genotype resistance testing at Chongqing Public Health Medical Center from May 2016 to June 2023. The HIV-1pol gene was amplified through RT-PCR and analyzed in terms of genotypic drug resistance. RESULTS: Of the 3015 PLWH tested for HIV-1 drug resistance, 1405 (46.6%) were resistant to at least one antiviral drug. Among non-nucleoside reverse transcriptase inhibitors (NNRTIs), 43.8% were resistant, compared to 29.5% for nucleoside reverse transcriptase inhibitors (NRTIs) and 3.4% for protease inhibitors (PIs). V179D/E and K103N/S were identified as the common mutation sites in the NNRTIs class of drugs, M184V/I and K65R/N were reported as the most common mutation sites in NRTIs, while thymidine analogue mutation (TAM) group was identified in 373 samples. L10FIV was the most common mutation in PIs. The dominant HIV-1 subtype was CRF07_BC. CONCLUSIONS: The high prevalence of HIV-1 drug resistance in Chongqing underscores the imperative for rigorous surveillance of the local HIV epidemic. Furthermore, TAMs are associated with HIV-1 multidrug resistance, and timely detection of drug resistance is helpful to reduce the emergence and spread of such drug-resistant strains.
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Fármacos Anti-VIH , Farmacorresistencia Viral , Genotipo , Infecciones por VIH , VIH-1 , Mutación , Humanos , VIH-1/genética , VIH-1/efectos de los fármacos , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , Infecciones por VIH/epidemiología , China/epidemiología , Farmacorresistencia Viral/genética , Estudios Retrospectivos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Fármacos Anti-VIH/uso terapéutico , Fármacos Anti-VIH/farmacología , Adulto Joven , Inhibidores de la Transcriptasa Inversa/uso terapéutico , Inhibidores de la Transcriptasa Inversa/farmacología , Adolescente , AncianoRESUMEN
This study aimed to determinate characteristics of drug resistance Mycobacterium tuberculosis from patients with extra-pulmonary tuberculosis (EPTB). Patients were retrospectively studied from January 2020 to December 2021. All the isolates were cultured, tested drug susceptibility, and detected the gene mutation using whole genome sequencing. The correlations of whole genome sequencing, pattern of DR, patients' distribution, and transmission were analyzed. 111 DR-EPTB isolates included pre-XDR-TB (53.2%), MDR-TB (29.7%), and poly-DR-TB (12.6%). The resistant drugs were INH followed by RFP and SM. The genotypes of 111 strains were lineage 2 and lineage 4. KatG_p.Ser315Thr was main gene mutation for resistance to INH; rpsL_p.Lys43Arg for SM, rpoB_p.Ser450Leu for rifampicin, embB_p.Met306Val for ethambutol, gyrA_p.Asp94Gly for FQs, and pncA_p.Thr76Pro for PZA. The residence was a significant risk factor for cluster transmission by patients and phenotypic DR types of strains for lineage 2 transmission. In the local area of southwest China INH, rifampicin and SM were main drugs in patients with DR-EPTB. KatG_p.Ser315, rpoB_p.Ser450Leu, and rpsL_p.Lys43Arg were main gene mutations. Phenotypic DR types and residence were main risk of transmission.
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Mycobacterium tuberculosis , Tuberculosis Extrapulmonar , Tuberculosis Resistente a Múltiples Medicamentos , Humanos , Mycobacterium tuberculosis/genética , Estudios Retrospectivos , Rifampin , Tuberculosis Resistente a Múltiples Medicamentos/genética , Secuenciación Completa del Genoma , Resistencia a MedicamentosRESUMEN
Objective To explore the structure and function of ß2 microglobulin-linker 2-human leukocyte antigen A24 (ß2m-linker 2-HLA A24) by bioinformatics, and to obtain the purified protein by eukaryotic expression. Methods The physicochemical properties of ß2m-linker 2-HLA A24 was predicted by ProtParam. The predicting software of phosphorylation site (NetPhos) and the predicting software of the secondary structure (SOPMA) were used to analyze the phosphorylation site and the secondary structure of the protein, respectively. And the predicting tool of modeling the proteins' structure (SWISS-MODEL) was used to model its tertiary structure. Then the recombinant plasmid of pcDNA3.4/ß2m-linker 2-HLA A24-His tag was constructed and transfected into Expi293F cells. The protein of interest was purified through Ni-affinity chromatography. And the purity and properties of the protein were identified by SDS-PAGE, Western blot analysis and indirect ELISA. Results The ß2m-linker 2-HLA A24 protein contained 44 phosphorylated sites. The protein was predicted to be a hydrophilic protein, of which the secondary structure mainly consists of irregular curl, and the modeling of the tertiary structure was successful. The recombinant plasmids of pcDNA3.4/ß2m-linker 2-HLA A24-His tag was successfully constructed and expressed in the Expi293F cells. Conclusion The ß2m-linker 2-HLA A24 protein has been successfully constructed. And bioinformatics results show that the protein has the function of binding to antigen peptides and activating T cells. It has established a foundation for activating antigen-specific CD8+ T cell stimulated by this protein and peptide in the subsequent experiments.
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Eucariontes , Antígeno HLA-A24 , Humanos , Péptidos/genética , Linfocitos T Citotóxicos/inmunologíaRESUMEN
OBJECTIVE: To investigate the pro-apoptotic effect and mechanism of miR-30a overexpression on chronic myeloid leukemia K562 cells. METHODS: The k562 cells were transfected with the recombinant plasmid pEGFP-pre-miR-30a, the real-time quantitative PCR was used to detect the level of miR-30a and BCR/ABL, and then the cell apoptosis was assessed by flow cytometry with AnnexinV-FITC/PI double staining. Western blot was used to detect the expression of BCR/ABL protein,apoptosis-related protein BCL-2 and BAX, PTEN, AKT and p-AKT. RESULTS: Sequencing and digestion map indicated that the recombinant plasmid was constructed successfully. Compared with 2 control groups, the miR-30a expression in k562 cells transfected with recombinant plasmid pEGFP-pre-miR-30a was obviously up-regulated. The expression of BCR/ABL mRNA and BCR/ABL protein was both significantly down-regulated. Apoptotic rate was significantly enhanced (both P<0.05),and the expression of anti-apoptotic protein BCL-2 was down-regulated while the expression of pro-apoptotic protein BAX was up-regulated. The level of PTEN was significantly up-regulated in omparison with control groups,no variation was found in total AKT, but the expression of p-AKT was down-regulated. CONCLUSION: The overexpression of miR-30a is abled to down-regulate the level of BCR/ABL mRNA and BCR/ABL protein, and increase apoptotic rate, its mechanism may be related with inhibition of the activity of BCR/ABL-PTEN/AKT signaling pathway.
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MicroARNs/genética , Apoptosis , Proliferación Celular , Proteínas de Fusión bcr-abl , Humanos , Células K562 , Leucemia Mielógena Crónica BCR-ABL PositivaRESUMEN
Objective To investigate the effect of short hairpin RNA (shRNA) knockdown of arachidonate 5-lipoxygenase (Alox5) gene on the apoptosis of resistant chronic myeloid leukemia K562/ADM cells. Methods Three pairs of shRNA fragment targeting human Alox5 gene were synthesized and inserted into pGenesil-1 interference vector. Enzyme digestion and sequencing were performed to identify the recombinant plasmid pGenesil-1-shRNA-Alox5. The plasmid was then transfected into K562/ADM cells. Real-time quantitative PCR and Western blotting were used to detect the Alox5 mRNA and protein levels to get the best interference group. The pGenesil-1-shRNA-Alox5 plasmid group with higher interference efficiency was selected as the experimental subjects. Real-time quantitative PCR was adopted to detect the expression level of bcr/abl mRNA and Western blotting to detect the BCR/ABL fusion protein, and the apoptotic rate was assessed by flow cytometry. Results The enzyme digestion and sequencing confirmed the successful construction of recombinant plasmid. Compared with the negative pGenesil-1-K562/ADM control group and blank K562/ADM cell group, the Alox5 mRNA and protein levels of K562/ADM cells transfected with pGenesil-1-shRNA-Alox5 recombinant plasmid significantly decreased, and after the knockdown of Alox5, the levels of bcr/abl mRNA and BCR/ABL fusion protein significantly decreased and the apoptosis rate increased obviously. Conclusion The knockdown of Alox5 gene can induce the decreased levels of bcl/abl mRNA and BCR/ABL fusion protein in the K562/ADM cells and increased apoptosis rate.