[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.

BACKGROUND:Previous research by the research team found that domestically produced porous tantalum is beneficial for early adhesion and proliferation of MG63 cells,and can be used as a scaffold material for bone tissue engineering. OBJECTIVE:To investigate the effect of domestic porous tantalum modified by osteogenic induction factor slow-release system on the adhesion,proliferation,and differentiation of MG63 cells. METHODS:Osteogenic induction factor slow-release system was constructed by adding 15%volume fraction of osteogenic factor solution to poly(lactic-co-glycolic-acid)gel.The passage 3 MG63 cells were inoculated on a porous tantalum surface(control group),porous tantalum surface coated with poly(lactic-co-glycolic-acid)copolymer gel(gel group),and porous tantalum surface coated with osteoblastic induction factor slow-release system(slow-release system group),and co-cultured for 5 days.The surface cytoskeleton of the material was observed by phalloidine staining.Cell proliferation was detected by flow cytometry.Western blot assay and RT-qPCR were used to detect the protein and mRNA expressions of type Ⅰ collagen,osteopontin,and RUNX-2 on the surface cells of the material. RESULTS AND CONCLUSION:(1)Phalloidine staining showed that MG63 cells adhered to and grew on the surface and inside of the three groups of porous tantalum,and the matrix secreted by the cells covered the surface of the material.(2)Flow cytometry showed that the cell proliferation in the slow-release system group was faster than that in the control group and the gel group(P<0.05).(3)Western blot assay and RT-qPCR showed that the protein and mRNA expressions of type Ⅰ collagen,osteopontin,and RUNX-2 in the slow-release system group were higher than those in the control group and gel group(P<0.05).(4)The results showed that the domestic porous tantalum modified by the osteogenic induction factor slow-release system was beneficial to the adhesion,proliferation,and differentiation of MG63 osteoblasts.

Objective:The incidence of early-onset colorectal cancer (EOCRC) is increasing globally; however, the molecular characteristics and prognosis of sporadic EOCRC are unclear. In this systematic review and meta-analysis, we aimed to investigate the incidence of gene mutations and their association with cancer survival in sporadic EOCRC, focusing on six common gene mutations ( TP53, BRAF, KRAS, NRAS, PTEN, and APC). Methods:Ovid Embase and Ovid Medline electronic databases were searched for studies involving patients with sporadic EOCRC (i.e., diagnosed with colorectal cancer before the age of 50 years and with no evidence of hereditary syndromes predisposing to colorectal cancer). The included articles were evaluated using quality assessment tools. Meta-analysis was performed using random-effects and fixed-effects models. Cochran's Q statistic and the I2 index were used to assess heterogeneity. The incidence of the six common gene mutations listed above in sporadic EOCRC and their association with cancer survival were evaluated.Results:(1) Incidence of specific gene mutations in sporadic EOCRC. A total of 34 articles were included in this meta-analysis. The incidence of APC gene mutation was 36% (from 13 articles, 95%CI: 19%-55%, P=0.043); of KRAS gene mutation 30% (from 26 articles, 95%CI: 24%-35%, P=0.190); of BRAF gene mutation 7% (from 18 articles, 95%CI: 5%-11%, P=0.422); of NRAS gene mutation 4% (from five articles, 95%CI: 3%-5%, P=0.586); of PTEN gene mutation 6% (from six articles, 95%CI: 4%-10%, P=0.968); and of TP53 gene mutation 59% (from 13 articles, 95%CI: 49%-68%, P=0.164). (2) Association between gene mutations and survival in sporadic EOCRC . A total of six articles were included in this meta-analysis. Compared with wild-type BRAF, mutant BRAF was significantly associated with increased overall mortality risk in patients with EOCRC (pooled HR=2.85, 95%CI: 1.45-5.60, P=0.002). Subgroup analysis showed that the incidence of BRAF gene mutation was higher in Eastern than in Western countries, whereas the incidence of TP53, KRAS, NRAS, and APC gene mutations was lower. There was no significant difference in the incidence of PTEN gene mutation between different regions. Conclusion:Compared with colorectal cancer occurring in the general population, the incidence of APC and KRAS mutations is lower in EOCRC, whereas the incidence of TP53 mutation remains consistent. BRAF mutation is associated with increased overall mortality risk in patients with EOCRC.

Objective:To investigate the effects of erythropoietin producing hepatocyte receptor A2 (EphA2) gene silencing on the growth, migration and invasion of human ovarian cancer HeyA8-MDR cells.Methods:After stable screening by plasmid and liposome mediated transfection, the expression of EphA2 protein was detected by Western blot. The effects of EphA2 gene silencing on the biological characters of HeyA8-MDR cells of ovarian cancer were observed by CCK-8 and Transwell chamber invasion assay.Results:Compared with HeyA8-MDR (negative control group) and HeyA8-MDR+ blank control group, the expression level of EphA2 protein in HeyA8-MDR+ experimental group was significantly decreased ( P<0.01). Cell proliferation, migration and invasion were inhibited ( P<0.05). Conclusions:EphA2 gene silencing can inhibit the growth, migration and invasion of HeyA8-MDR cells of ovarian cancer, providing a new idea for the treatment of ovarian cancer.

Objective:The incidence of early-onset colorectal cancer (EOCRC) is increasing globally; however, the molecular characteristics and prognosis of sporadic EOCRC are unclear. In this systematic review and meta-analysis, we aimed to investigate the incidence of gene mutations and their association with cancer survival in sporadic EOCRC, focusing on six common gene mutations ( TP53, BRAF, KRAS, NRAS, PTEN, and APC). Methods:Ovid Embase and Ovid Medline electronic databases were searched for studies involving patients with sporadic EOCRC (i.e., diagnosed with colorectal cancer before the age of 50 years and with no evidence of hereditary syndromes predisposing to colorectal cancer). The included articles were evaluated using quality assessment tools. Meta-analysis was performed using random-effects and fixed-effects models. Cochran's Q statistic and the I2 index were used to assess heterogeneity. The incidence of the six common gene mutations listed above in sporadic EOCRC and their association with cancer survival were evaluated.Results:(1) Incidence of specific gene mutations in sporadic EOCRC. A total of 34 articles were included in this meta-analysis. The incidence of APC gene mutation was 36% (from 13 articles, 95%CI: 19%-55%, P=0.043); of KRAS gene mutation 30% (from 26 articles, 95%CI: 24%-35%, P=0.190); of BRAF gene mutation 7% (from 18 articles, 95%CI: 5%-11%, P=0.422); of NRAS gene mutation 4% (from five articles, 95%CI: 3%-5%, P=0.586); of PTEN gene mutation 6% (from six articles, 95%CI: 4%-10%, P=0.968); and of TP53 gene mutation 59% (from 13 articles, 95%CI: 49%-68%, P=0.164). (2) Association between gene mutations and survival in sporadic EOCRC . A total of six articles were included in this meta-analysis. Compared with wild-type BRAF, mutant BRAF was significantly associated with increased overall mortality risk in patients with EOCRC (pooled HR=2.85, 95%CI: 1.45-5.60, P=0.002). Subgroup analysis showed that the incidence of BRAF gene mutation was higher in Eastern than in Western countries, whereas the incidence of TP53, KRAS, NRAS, and APC gene mutations was lower. There was no significant difference in the incidence of PTEN gene mutation between different regions. Conclusion:Compared with colorectal cancer occurring in the general population, the incidence of APC and KRAS mutations is lower in EOCRC, whereas the incidence of TP53 mutation remains consistent. BRAF mutation is associated with increased overall mortality risk in patients with EOCRC.

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.

Abstract Improving health literacy can effectively reduce the risk of health risk behaviors and adverse health outcomes in children and adolescents. Global research on health literacy of children and adolescents has been increasing in recent years. The paper reviews the existing research and proposes that there is no universal definition and connotation of health literacy for children and adolescents, most of the studies use or make some revisions on the definition of adult health literacy in child and adolescent health literacy, failing to consider developmental characteristics and unique health needs of children and adolescents. Moreover, the assessment index system and instruments are diversified, making the research findings from different studies difficult to compare, and to obtain consistent and reliable conclusions. Future endeavors are encoouraged to expand health literacy researches and to update more comprehensive and practical definition, and to develop a standardized assessment instrument that can be validated in Chinese culture.

Objective:To investigate the application of serum creatinine to prealbumin ratio (Scr/PA) in the diagnosis of patients with heart failure complicated with renal failure.Methods:This was a case-control study. Patients with chronic heart failure and heart failure complicated with renal failure admitted to Dalian Central Hospital from January 5, 2020 to April 23, 2023 were retrospectively analyzed, and Scr/PA was calculated. The general data and laboratory examination indexes of the two groups were compared. According to the data type, t test, Wilcoxon rank sum test and χ 2 test were used for comparison between the two groups. The risk factors of heart failure complicated with renal failure were analyzed by univariate and multivariate logistic regression analysis, and Spearman correlation analysis was used to analyze the correlation between Scr/PA and N-terminal pro-B-type natriuretic peptide (NT-proBNP) and hemoglobin (HGB). ROC curve was used to determine the predictive value of Scr/PA and NT-proBNP for heart failure complicated with renal failure. Results:Compared with the heart failure group, Triglyceride [1.25 (0.94, 1.81) mmol/L vs. 1.07 (0.76, 1.46) mmol/L, Z=-2.159, P=0.031], D-dimer [2.30 (1.53, 4.67) mg/L vs. 1.63 (0.64, 2.96) mg/L, Z=-2.339, P=0.02],NT-proBNP [18 500 (9 575, 30 000) pg/ml vs. 4 865 (1 600, 9 800) pg/ml, Z=-5.637, P<0.001], Scr/PA [0.233 (0.188, 0.351) mg/mg vs 0.064 (0.044, 0.103) mg/mg, Z=-8.197, P<0.001] were higher in heart failure complicated with renal failure group. While albumin [(33.9±5.2) g/L vs. (36.3±4.3) g/L, t=-2.173, P=0.008], estimated glomerular filtration rate[12.86 (7.88, 17.40) ml/(1 min×1.73 m 2) vs. 65.82 (48.66, 86.32) ml/(1 min×1.73 m 2), Z=-9.794, P<0.001], and HGB [(91±24) g/L vs. (123±23) g/L, t=-7.489, P<0.001] were lower. Univariate logistic regression analysis showed that albumin ( OR=0.900, 95% CI 0.830-0.975, P=0.010), HGB ( OR=0.948, 95% CI 0.930-0.966, P<0.001), Scr/PA ( OR=1.639, 95% CI 1.346-1.957, P<0.001) were associated with heart failure complicated with renal failure. Multivariate logistic regression analysis showed that only Scr/PA was an independent risk factor for heart failure complicated with renal failure. The correlation coefficients of Scr/PA with NT-proBNP and HGB were r=0.578 and r=-0.559, respectively (all P<0.001). The area under the AUC curve of Scr/PA and NT-proBNP for predicting heart failure complicated with renal failure was 0.927 (95% CI: 0.881-0.973, P<0.001) and 0.797 (95% CI: 0.717-0.877, P<0.001), respectively. Conclusions:Scr/PA is an independent risk factor for heart failure complicated with renal failure, and it has a good correlation with NT-proBNP and HGB. Scr/PA is superior to NT-proBNP in predicting heart failure complicated with renal failure.

Objective: To understand the current situation regarding sex education in home and school settings in China, and to provide a scientific basis for more effective family and school sex education. Methods: Using a convenient sampling method, a crosssectional survey of students from grade 7 to grade 9 in Beijing, Liaoning, Sichuan, Chongqing, Yunnan and Henan was carried out from September to December 2021. The questionnaire covered general demographic information, sex education at home and school, and the status of comprehensive sexuality education. Finally, 4 109 valid questionnaires were collected. Results: A total of 760(18.50%) middle school students had not received any form of sex education, 923(22.46%) had only received sex education at school, 299(7.28%) had only received sex education at home, and 2 127 (51.76%) had received sex education at home and at school. The scores related to junior middle school students knowledge of comprehensive sexuality education was (12.43±2.89) points, the attitude score was (47.86±5.31) points, the behavior score was (40.44±5.18) points, and the comprehensive level score was (82.02±9.01) points. Junior middle school students who received sex education at home and in school had the highest comprehensive level of sex education( P <0.01). Junior middle school students who only received sex education at school had a higher comprehensive sexuality education knowledge level than those who only received sex education at home( P < 0.01 ). The level of comprehensive sexuality education behavior among junior middle school students who only received family sex education was significantly higher than among those who only received sex education at school( P <0.01). Conclusion Receiving sex education in both home and school settings can effectively improve the comprehensive sexuality education competency among junior middle school students. School based sex education should be further implemented in the future and family sex education should be encouraged.

Improper control of depth of anesthesia is not only detrimental to the rapid and stable recovery of anesthesia, but also affects the postoperative outcome of patients. Therefore, accurate control of anesthesia depth is an urgent clinical and scientific problem in the field of anesthesiology. At present, different algorithm models derived from electroencephalogram (EEG) signals are used to monitor the depth of anesthesia, but they cannot meet the requirements of anesthesiologists to accurately evaluate the depth of anesthesia. In recent years, the research on the mechanism and modulation of anesthesia-related neural network suggests that it has potential value as a method to monitor depth of anesthesia. Anesthesia-related neural networks mainly include sleep-wake circuit, thalamic-cortical circuit and corticocortical network. A thorough understanding of the neural network involved in the loss of consciousness caused by anesthesia will guide the depth of anesthesia monitoring more accurately and provide possibility for improving the quality of clinical anesthesia resuscitation.