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[Objective] Genetic polymorphisms determine individual genetic differences and disease susceptibility and affect drug metabolism and transport. Genetic polymorphism testing has significant application value in various fields, such as medical research, disease diagnosis, prevention, and health management. Fluorescence quantitative polymerase chain reaction (qPCR) instruments are highly sensitive and accurate molecular biology detection tools widely used in basic medical research and clinical diagnosis. Thus, medical students need to learn the fluorescence qPCR technique to improve their research capabilities and disease diagnosis levels and respond to public health events. This experiment designs three methods, namely high-resolution melting curve (HRM), allele-specific PCR (AS-PCR), and TaqMan probe, to genotype the ethanol dehydrogenase 1B (ADH1B) rs1229984 locus, aiming to guide students in using different PCR-derived techniques for genetic polymorphism testing. [Methods] The ADH1B gene sequence and the homologous genes ADH1C and ADH1A with high homology were downloaded from the GenBank database. The SnapGene software for gene alignment was used to locate the rs1229984 locus. Amplification primers for HRM-PCR, AS-PCR, and direct sequencing were designed using software such as SNPway, Primer, and Beacon Designer and algorithms such as NUPACK and MetLab. Commercial TaqMan probe primer pairs were ordered. Oral epithelial cells were successively collected from 48 volunteers, DNA was extracted using a rapid extraction method, and DNA content was measured using a spectrophotometer. PCR amplification systems were configured for the HRM, AS, and TaqMan methods, amplification reactions were performed on the corresponding PCR instruments, and the corresponding software was used for result analysis after the reaction ended. At the same time, the PCR products were detected using Sanger sequencing to identify the genotype of ADH1B rs 1229984. [Results] The HRM, AS, and TaqMan methods can all accurately identify the polymorphism of the ADH1B gene, and their genotyping results are consistent with the results of the direct and Sanger sequencing methods. Each of the three PCR methods has its advantages and disadvantages: The TaqMan method has strong sensitivity and specificity but requires designing a pair of primers in addition to a pair of locus-specific probes, which is costly. The HRM method is the simplest to operate and the lowest in cost, but it requires high-quality DNA. The AS method has strong specificity but requires high primer design standards and involves PCR amplification reactions with two upstream primers and one downstream primer, comparing the ΔCt values of the two for judgment, which is relatively complex. [Conclusions] By establishing the HRM, AS, and TaqMan methods, the polymorphism of the ADH1B gene rs122998 locus in 48 volunteers was successfully genotyped. The accuracy of the experimental results confirmed the reliability of fluorescence qPCR technology in the analysis of genetic polymorphisms. Through experimental teaching, students can be taught to apply what they have learned to detect other genetic polymorphisms. When choosing a detection method, several factors, such as the purpose of the experiment, number of samples, cost, and ease of operation, need to be considered comprehensively. [ABSTRACT FROM AUTHOR]

This retrospective cohort study was conducted to explore the effect of traditional Chinese medicine (TCM) therapy on the long-term trends in CD4+ T-cell count among patients with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) who were treated with combined antiretroviral therapy (cART) over a 14-year period. A total of 721 individuals were treated with cART alone (cART group), and 307 individuals were treated with both cART and TCM (TCM + cART group). Among all enrolled patients with HIV/AIDS, 99.5% were farmers, 71.1% had more than 6 years of education, and 96.8% were infected with HIV via a paid blood donation. For those patients with HIV/AIDS who had a baseline CD4+ T-cell count of <350 cells/mL, the CD4+ T-cell count tended to increase to approximately 350 cells/mL more rapidly in the TCM + cART group than in the cART group, but when the baseline CD4+ T-cell count was ≥350 cells/mL, there was no difference between the cART and TCM + cART groups. For other patients with HIV/AIDS who had a baseline CD4+ T-cell count of 350–500 cells/mL, the CD4+ T-cell counts tended to increase slightly, but there was no difference between the two groups. For patients with HIV/AIDS who had a baseline CD4+ T-cell count of ≥500 cells/mL, the CD4+ T-cell counts tended to be maintained at a particular level, with no difference between the two groups. The results show that the effect of TCM on the CD4+ T-cell counts of patients with HIV/AIDS is related to the CD4+ T-cell level at the time of initial treatment. TCM can increase the CD4+ T-cell count among patients with HIV/AIDS who have a baseline CD4+ T-cell count of <350 cells/mL. Sex and age have a slight influence on the therapeutic effect of TCM. [ABSTRACT FROM AUTHOR]

Abstract Background High circulating luteinizing hormone (LH) level is a typical biochemical feature of polycystic ovary syndrome (PCOS) whose pathophysiology is still unclear. Certain mutations of LH and LH receptor (LHR) may lead to changes in bioactivity of these hormones. The aim of this study was determine the role of the LH and LHR polymorphisms in the pathogenesis of PCOS using a genetic approach. Methods 315 PCOS women and 212 controls were screened for the gene variants of LH G1052A and LHR rs61996318 polymorphisms by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Results PCOS patients had significantly more A allele frequency of LH G1052A mutations than controls (p=0.001). Within PCOS group, carriers of LH 1052A allele had lower LH (p=0.05) and higher fasting glucose levels (p=0.04). No subjects were identified with LHR rs61996318 polymorphisms. A new LHR single nucleotide polymorphism (SNP) was found without clear association with PCOS. Conclusions Results suggested LH G1052A mutation might influence PCOS susceptibility and phenotypes.

Abstract Background Tribbles 3 (TRB3) affects insulin signalling by inhibiting insulin-stimulated Akt phosphorylation and subsequent activation. A single nucleotide polymorphism located in the second extron of the human TRB3 gene is thought to be associated with insulin resistance. The latter is a core abnormality in PCOS independent of obesity. The present study was designed to clarify the relationships of TRB3 Q84R polymorphism with PCOS in a Chinese women group. Methods A case-control study with two groups: PCOS group (n = 336) and control group of infertility women for tubal and/or male factor (n = 116) was performed. Genotyping of the TRB3 R84 variant was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results The frequency of genotype QQ in PCOS women was significantly lower, while genotype QR and RR were significantly higher than that in control group (p < 0.05). However, the difference disappeared after adjustment for BMI. At glucose1h, glucose2h and insulin2h point, the difference between QQ individuals and R84 allele carriers in PCOS women reached statistical significance during OGTT (p < 0.05). Conclusions TRB3 Q84R polymorphism is associated with obesity and especially glucose metabolism and not associated with polycystic ovary syndrome because of compositional characteristics of phenotype in Chinese PCOS women.

The conventional survival analysis model on HIV/AIDS prognosis is the Cox proportional hazard model, which deals with only one event type, death, regardless of the cause. Few studies have used a competing risk model to evaluate the predictors of AIDS-related mortality. To estimate the influence of antiretroviral therapy (ART) initiation time and baseline CD4+ cell counts on acquired immunodeficiency syndrome (AIDS)-related death among former plasma donors. A retrospective cohort study was conducted involving 11,905 human immunodeficiency virus (HIV) or AIDS patients in a high-risk area of Henan province in China between 1995 and 2016. Demographic and clinical data were collected. Sub-distribution hazard ratios (sHRs) for AIDS-related mortality with baseline CD4+ cell counts and ART initiation time were determined using a competing risk model. Patients who initiated ART within 90 days of HIV/AIDS diagnosis (sHR: 0.24, 95% CI: 0.22–0.27) or had baseline CD4+ counts of >500 cells/μL (sHR: 0.23, 95% CI: 0.19–0.28) were associated with lower AIDS-related mortality risk. Patients with ART initiation time >1 year but CD4+ counts >350 cells/μL (sHR: 4.42, 95% CI: 3.30–5.91) had a higher AIDS-related mortality risk than those with ART initiation time >90 days but CD4+ counts ≤350 cells/μL (sHR: 4.33, 95% CI: 3.58–5.23). Our results demonstrate that patients with high CD4+ cell counts and late ART had a 9% higher risk of AIDS-related death than those with low CD4+ cell counts and early ART. This study confirms the great significance of immediate ART initiation among former plasma donor HIV patients in China. [ABSTRACT FROM AUTHOR]

The Ca3CoMn1-xTixO6(x = 0, 0.05) samples were synthesized by a modified sol-gel method, and the microstructure, microtopography, and magnetic properties were investigated in detail. With substituting of Mn4+ ions by Ti4+ ions, the crystallinity and particle size of Ca3CoMn1-xTixO6 (x = 0, 0.05) samples decrease. The Ti4+ doping also causes the high-temperature spin freezing phenomenon disappears, and the transition field of ↑↑↓↓ magnetic structure to ↑↑↑↓ magnetic structure increases obviously together with the faster dynamic behavior. These results suggest that the dilution effect of Ti4+ doping to Co-Mn-Co-Mn spin chains could reduce the degree of magnetic frustration and suppress the formation of short range magnetic order, for which will be beneficial for the establishment of more robust long range antiferromagnetic structure. The present research could serve as a reference on tuning the magnetic order and improving the multiferroic property of Ca3CoMnO6. [ABSTRACT FROM AUTHOR]