[Objective] To compare next generation sequencing (NGS) library construction technology between probe hybridization capture and amplicon methods, and analyze the influencing factors of HLA genotyping resolution level and its prospects in clinical applications. [Methods] A total of 207 clinical samples with known typing results and samples from the proficiency testing plan were selected. The conformity rate of HLA genotyping results, allele coverage and typing data analysis indicators were confirmed, and the effects of two library construction methods on the level of HLA genotyping discrimination were compared. [Results] The concordance rate of 207 samples with the feedback results of PT or prior well-characterized HLA genotypes was 100%. Among them, 91 samples were captured using hybridization probe capture method. Compared with the original amplicon method, the hybridization probe capture method can distinguish the alleles of DRB1 and DPB1 that cannot be determined in 13 samples. The allelic imbalance of DRB1, DPA1, and DQB1 loci in 6 samples was resolved. Three samples were found to have missed detection of alleles at the DQA1 and DQB1 loci. [Conclusion] The performance indicators of hybridization probe capture and amplicon performance confirmation meet the requirements of clinical detection of HLA genotyping, which provides an experimental method and basis for clinical application.

Objective　By exploring the function of sulfakinin (SK) and sulfakinin receptor (SKR) of Aedes aegypti, it laid a certain experimental basis and theoretical basis for the research and development of new insecticides targeting neuropeptides and their receptors. Methods　This study investigated the roles of SK and its receptor gene in Ae. aegypti using bioinformatics analysis and Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/Cas9 knockout technology. Subsequently, RNA interference technology was employed to suppress the expression of SK or its receptor in adult mosquitoes. Lastly, transcriptome sequencing technology was utilized to identify and analyze differentially expressed genes between the interference group and the control group in order to gain insights into their functions. Results　It was found that there is only one SK receptor in Ae. aegypti. In addition, during the construction of mutant strains of Ae. aegypti SK and its receptor gene, it was found that only 2% of the G0 generation mutant strains mutated to form chimeras, with a large number of male chimeras dying, and only 14% of female chimeras being able to lay eggs, ultimately resulting in no effective G1 generation mutants. Transcriptome data showed, compared to the control group, 181 genes were significantly differentially expressed after interfering with the SK gene, with 62 genes significantly upregulated and 119 genes significantly downregulated. In addition, after interference with the sulfakinin receptor, 110 genes exhibited significant differential expression, including 20 upregulated and 90 downregulated genes. Cross-analysis of the two datasets identified 46 genes with significant expression changes after interference with sulfakinin or its receptor, with only 4 genes upregulated and the remaining 42 genes significantly downregulated, and the differentially expressed genes were mainly enriched in the metabolic pathway, endocrine system, and digestive system. Conclusions　The SK and its receptor gene are highly conserved and may primarily play roles in regulating the energy metabolism and digestion functions in Ae. aegypti, thus playing an important role in regulating insect growth and development.

Objective:To investigate the role and related mechanisms of the LiaSR two-component system in acid tolerance and biofilm formation abilities of Streptococcus mutans (Sm) 593. Methods:The growth curves of various Sm strains in pH=5.5 brian heart infusion (BHI) medium were analyzed. And colony forming unit (CFU) was also performed to evaluate the acid tolerance of Sm. Laurdan probe, H +-K +adenosine triphosphate (ATP)ase activity analysis kit, proton permeability assay and real-time fluorescence quantitative PCR (RT-qPCR) were conducted to detect the acid tolerant mechanisms of LiaSR two-component system in Sm. Crystal violet staining, CFU, SYTOX probe and anthrone-sulfuric method were used to analyze the properties and structures of the Sm biofilms. RT-qPCR was conducted to detect the expression levels of underlying regulated genes. Results:The growth of mutants in acidic BHI were inhibited ( P<0.05). The acid tolerance of mutants significantly decreased compared to the wild-type strain ( P<0.05). In mutants, the activity of H +-ATPase (917.06±59.53 and 469.53±47.65) were elevated by 7.22-folds and 3.70-folds compared to the wild-type strain (127.00±50.71) ( P<0.001, P<0.001) and the encoded gene atpD (3.39±0.21 and 1.94±0.17) were also elevated by 3.39-folds and 1.94-folds compared to the wild-type strain (1.00±0.15) ( P<0.001, P=0.001). The Laurdan generalized polarization of mutants (0.18±0.04 and 0.18±0.05) increased significantly compared to the wild-type strain (0.08±0.05) ( P=0.006, P=0.003) and the expression levels of fabM gene were decreased in mutants (0.52±0.11 and 0.57±0.05) by 1/2 ( P=0.014, P=0.022). In liaR deletion mutant, the reduced terminal pH (4.76±0.01) can also be observed ( P<0.001). The total amount of the biofilms of three Sm didn't show significant differences ( P>0.05). But the number of viable bacteria of mutants′ biofilms were decreased [Sm 593: (12.00±2.80)×10 7 CFU/ml; Sm ΔliaS: (2.95±1.13)×10 7 CFU/ml; Sm ΔliaR: (7.25±1.60)×10 7 CFU/ml] ( P=0.001, P=0.024). The extracellular DNA were increased by 18.00-folds and 6.50-folds in mutants′ biofilms (128.73±15.65 and 46.38±5.52) compared to the wild-type strain (7.16±3.62) ( P<0.001, P=0.003). Water-soluble exopolysaccharides could be found up-regulated in liaS deletion mutant [(138.73±10.12) μg/ml] ( P=0.003) along with the expression level of gtfC gene (1.65±0.39) ( P=0.014). The expression level of gtfD were elevated by 47.43-folds and 16.90-folds in mutants ( P<0.001, P=0.010). Conclusions:The LiaSR two-component system can promote the expression of fabM gene and increase the fluidity of Sm which contributes to acid tolerance. The LiaR can also decrease the proton permeability and restrict the entrance of H +. The LiaSR two-component system can negatively regulate the production of the extracellular matrix in Sm biofilm.

Objective To explore the molecular mechanism by which FEZF1-AS1 overexpression promotes progression of non-small cell lung cancer(NSCLC)via the miR-130a-5p/CCND1 axis.Methods TCGA database was used to analyze FEZF1-AS1 expression levels in NSCLC.FEZF1-AS1 expression was detected by qRT-PCR in clinical specimens of NSCLC tissues and NSCLC cell lines,and its correlation with clinical features of the patients were analyzed.The binding sites of FEZF1-AS1 with hsa-miR-130a-5p and those of hsa-miR-130a-5p with CCND1 were predicted.CCK8 assay,clone formation assay,scratch assay,and Transwell assay were employed to examine the effects of FEZF1-AS1 knockdown and hsa-miR-130a-5p inhibitor on proliferation,invasion,and migration abilities of lung cancer cell lines.Dual luciferase assay was used to verify the binding of FEZF1-AS1 with hsa-miR-130a-5p and the binding of hsa-miR-130a-5p with CCND1.Western blotting was performed to detect the changes in CCND1 protein expression level in H1299 and H358 cells following FEZF1-AS1 knockdown and treatment with hsa-miR-130a-5p inhibitor.Results FEZF1-AS1 was highly expressed in NSCLC tissues in close correlation with lymph node metastasis and also in H1299 and H358 cell lines(all P<0.05).FEZF1-AS1 knockdown obviously reduced proliferation,migration,and invasion abilities of NSCLC cells(P<0.05).Dual luciferase assay confirmed the binding of hsa-miR-130a-5p with FEZF1-AS1 and CCND1(P<0.05),and hsa-miR-130a-5p inhibitor significantly inhibited proliferation,migration,and invasion of NSCLC cells(P<0.05).FEZF1-AS1 knockdown significantly reduced CCND1 protein expression in NSCLC cells,and this effect was strongly inhibited by treatment with hsa-miR-130a-5p inhibitor(P<0.05).Conclusion FEZF1-AS1 is highly expressed in NSCLC tissue in close correlation with lymph node metastasis to promote cancer progression through the miR-130a-5p/CCND1 axis.

Objective To explore the molecular mechanism by which FEZF1-AS1 overexpression promotes progression of non-small cell lung cancer(NSCLC)via the miR-130a-5p/CCND1 axis.Methods TCGA database was used to analyze FEZF1-AS1 expression levels in NSCLC.FEZF1-AS1 expression was detected by qRT-PCR in clinical specimens of NSCLC tissues and NSCLC cell lines,and its correlation with clinical features of the patients were analyzed.The binding sites of FEZF1-AS1 with hsa-miR-130a-5p and those of hsa-miR-130a-5p with CCND1 were predicted.CCK8 assay,clone formation assay,scratch assay,and Transwell assay were employed to examine the effects of FEZF1-AS1 knockdown and hsa-miR-130a-5p inhibitor on proliferation,invasion,and migration abilities of lung cancer cell lines.Dual luciferase assay was used to verify the binding of FEZF1-AS1 with hsa-miR-130a-5p and the binding of hsa-miR-130a-5p with CCND1.Western blotting was performed to detect the changes in CCND1 protein expression level in H1299 and H358 cells following FEZF1-AS1 knockdown and treatment with hsa-miR-130a-5p inhibitor.Results FEZF1-AS1 was highly expressed in NSCLC tissues in close correlation with lymph node metastasis and also in H1299 and H358 cell lines(all P<0.05).FEZF1-AS1 knockdown obviously reduced proliferation,migration,and invasion abilities of NSCLC cells(P<0.05).Dual luciferase assay confirmed the binding of hsa-miR-130a-5p with FEZF1-AS1 and CCND1(P<0.05),and hsa-miR-130a-5p inhibitor significantly inhibited proliferation,migration,and invasion of NSCLC cells(P<0.05).FEZF1-AS1 knockdown significantly reduced CCND1 protein expression in NSCLC cells,and this effect was strongly inhibited by treatment with hsa-miR-130a-5p inhibitor(P<0.05).Conclusion FEZF1-AS1 is highly expressed in NSCLC tissue in close correlation with lymph node metastasis to promote cancer progression through the miR-130a-5p/CCND1 axis.

Background::Limited information exists regarding the impact of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection on psoriasis patients. The objective of this study was to identify clinical factors associated with the prognosis of psoriasis following SARS-CoV-2 infection.Methods::A retrospective, multicenter study was conducted between March and May 2023. Univariable and multivariable logistic regression analyses were employed to identify factors associated with coronavirus disease 2019 (COVID-19)-related psoriasis outcomes. The study included 2371 psoriasis patients from 12 clinical centers, with 2049 of them having been infected with SARS-CoV-2.Results::Among the infected groups, lower exacerbation rates were observed in individuals treated with biologics compared to those receiving traditional systemic or nonsystemic treatments (22.3% [236/1058] vs. 39.8% [92/231] vs. 37.5% [140/373], P <0.001). Psoriasis progression with lesions (adjusted odds ratio [OR] = 8.197, 95% confidence interval [95% CI] = 5.685–11.820, compared to no lesions), hypertension (adjusted OR = 1.582, 95% CI = 1.068–2.343), traditional systemic (adjusted OR = 1.887, 95% CI= 1.263–2.818), and nonsystemic treatment (adjusted OR= 1.602, 95% CI= 1.117–2.297) were found to be associated with exacerbation of psoriasis after SARS-CoV-2 infection, but not biologics (adjusted OR = 0.931, 95% CI = 0.680–1.274, compared to no treatment), according to multivariable logistic regression analysis. Conclusions::A reduced risk of psoriasis exacerbation after SARS-CoV-2 infection was observed with biologics compared to traditional systemic and nonsystemic treatments. Significant risk factors for exacerbation after infection were identified as existing psoriatic lesions and hypertension.

Objective:To establish a linkage prediction model for human leukocyte antigen (HLA) DPA1-DPB1 and validate it by using clinical data and follow-up data from unrelated allogeneic hematopoietic stem cell transplantation donors and recipients, and to explore the clinical application value of the prediction model in transplantation prognosis.Methods:This is a retrospective study. Leveraging the artificial neural network algorithm of NetMHCⅡpan and the DPA1-DPB1 haplotype linkage database of the Chinese population established in our previous research, and incorporating the amino acid FASTA data of DPA1-DPB1 of all known sequences newly published by the Latest International Immunogenetics/Human Leukocyte Antigens, 47 DPA1-DPB1 linkage models were established. Employing next-generation sequencing technology based on the hybridization capture library construction method, HLA genotyping tests for HLA-A, -B, -C, DRB1, DQB1, DQA1, DRB3/4/5, DPB1, and DPA1 (9 loci) were performed on 250 donor-recipients pairs who underwent unrelated-donor hematopoietic stem cell transplantation in the Department of Hematology of the First Affiliated Hospital of Soochow University between January 2016 and September 2021. HLA typing data and clinical information of transplant donors and recipients were retrospectively analyzed to assess and predict the impact of permissive and non-permissive linkage mismatches of DPA1-DPB1 on transplantation prognosis. The Kaplan-Meier method with the log-rank test was applied to compare the survival curves of overall survival (OS) rates between different groups. Additionally, a competing risks model was utilized to compare the cumulative incidence of grade Ⅱ-Ⅳ acute graft-versus-host disease and non-relapse mortality (NRM) across groups. The area under the receiver operating characteristic curve was employed to compare the predictive performance of the established prediction model with that of the T-cell epitope (TCE) model.Results:According to the different hydrophilic and hydrophobic properties of amino acids, the DPA1-DPB1 linkage model is categorized into types Ⅰ-Ⅳ: type I consists of 6 hydrophobic types at P1-P8 plus hydrophilic type at P9; type Ⅱ includes 17 hydrophobic types; type Ⅲ comprises 9 amphiphilic types; and type Ⅳ consists of 15 hydrophilic types. According to the prediction model, DPA1-matched and DPB1-mismatched donor-recipient cases were classed into P1-matched or P1-mismatched groups. Compared with fully matched DPA1 and DPB1 cases, P1-mismatched patients had a 2-year OS rate of 75% (12/16) versus 96.2%(25/26) (χ2=4.13, P=0.04), and a NRM rate of 4/16 versus 0 (χ2=7.05, P<0.01). However, there was no statistically significant difference in the 2-year OS and NRM rates compared to DPA1 and DPB1 cases ( P>0.05). The prediction model established in this study demonstrated a larger area under the receiver operating characteristic curve for predicting the 2-year OS rate compared with the DPB1 TCE model ( Z=0.71, P=0.48). In donor-recipient cases where both DPA1 and DPB1 were mismatched, the 2-year OS rates decreased and the NRM increased in both P1-matched and P1-mismatched cases compared with fully matched DPA1 and DPB1. Moreover, P1-mismatched patients had a worse prognosis compared to P1-matched patients. Conclusion:The DPA1-DPB1 linkage prediction model established based on high-throughput next-generation sequencing technology can be used to predict the impact of HLA-DP mismatches on OS and NRM in transplantation, and the prediction performance is superior to the TCE model.

Objective To evaluate feasibility of early skeletal muscle injury by using shear wave elastography.Methods Sixty patients with unilateral knee osteoarthritis treated in Department of Orthopedics of People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine from May 2020 to January 2021 were selected as the research subjects.Conventional scanning and shear wave elastography techniques were applied to investigate affected and healthy knees.Measurement included cartilage morphology,thickness and damage,joint cavity effusion,synovial membrane thickness,ligament morphology,meniscus protrusion degree,and detection of shear wave values in attachment area of quadriceps tendon on patella.Thickness and shear wave values of tendons in different observation areas on affected side and healthy side were compared.Affected knee joint was assigned a value based on the ultrasound characteristics,and Spearman correlation was used to analyze correlation between ultrasound score and shear wave value.Results Among 60 patients,fifty-six cases(56/60,93.3%)of cartilage injury in knee joint,30 cases(30/60,50.0%)of synovial hyperplasia in knee joint,35 cases(35/60,53.3%)of medial meniscus extrusion in knee joint,57 cases(57/60,95.0%)of effusion in knee joint cavity,21 cases(21/60,35%)of osteophyte formation in knee joint,and zero case of popliteal cyst in knee joint.There were 34 cases with ultrasound score less than three points,26 cases with score three to seven points,and zero case with score more than seven points.Shear wave value of the tendon in affected lateral area was significantly greater than that in affected medial area(P<0.01).Shear wave value of tendon in healthy medial area was significantly higher than that in affected medial area(P<0.01).Shear wave value of quadriceps tendon decreases with increase of ultrasound score,showed a high correlation(r=0.955,P<0.01).Conclusion Shear wave elastography is a non-invasive and relatively easy-to-operate method for evaluating muscle injury,which is highly correlated with severity of knee osteoarthritis,can help detect early strain points,providing possibilities for grading and diagnosis of knee osteoarthritis.

Objective:To evaluate the effectiveness of early predictive intervention based on the LACEinde (LACE) in aged patients undergoing surgery for non-small cell lung cancer (NSCLC) .Methods:Totally 116 aged NSCLC patients who underwent surgical treatment at the First Affiliated Hospital of Zhengzhou University from March 2021 to March 2023 were selected by convenience sampling. They were randomly divided into a control group and a study group, with 58 patients in each. Patients in the control group received routine care, while patients in the study group underwent early predictive intervention based on LACE in addition to the routine care. Respiratory function, cancer-related fatigue, quality of life, and prognosis of patients in both groups were assessed before and after the intervention.Results:After the intervention, the study group showed significantly higher values in vital capacity (VC) and forced vital capacity (FVC) compared to the control group ( P<0.01). The Piper Fatigue Scores (PFS) and the Quality of Life Questionnaire-Lung Cancer 13 (QLQ-LC13) scores in the study group were significantly lower than those in the control group after the intervention ( P<0.01). The rates of unplanned readmission and total complications in the study group were significantly lower than those in the control group ( P<0.05) . Conclusions:Early predictive intervention based on LACE can reduce the rate of unplanned readmissions and complications, improve respiratory function, reduce cancer-related fatigue, and enhance the quality of life in aged NSCLC patients post-surgery.