ObjectiveThis study aims to explore the potential of different orders of magnitude single-nucleotide polymorphism (SNP) locus combinations for predicting distant kinship relationships. A high-density SNP locus set was constructed, and a comprehensive assessment of its inference capability was conducted. MethodsFirstly, we selected three commercial chip panels, CGA (Chinese genotyping array, Illumina), GSA (Global screening array, Illumina), Affy (23MF_V2 high-density SNP array, Affymetrix) and merged them after quality control, forming a high-density SNP locus panel(1 180 k). Secondly, we selected 161 samples and collected their peripheral blood samples by using whole-genome sequencing technology. Within this sample population, the levels of kinship relationships fully covered the range from level 1 to level 9, and the number of kinship pairs at each level was consistently maintained at over 50 pairs. From 161 samples data of whole-genome sequencing, the 1 180 k locus set was extracted, which is referred to as the high-density SNP locus set in the following text. The kinship inference was conducted using the identity-by-descent (IBD) algorithm with the selected optimal parameters. To comprehensively evaluate the performance of the high-density SNP locus set in kinship inference, we compared it with the three commercial chip panels, the intersection of these three chip loci, and the control sets constructed by randomly reducing the number of the high-density SNP locus set. Based on the changes in the IBD lengths, as well as the dynamic trends in prediction accuracy, we conducted a scientific assessment of the kinship inference capability of the high-density SNP locus set. ResultsAfter screening, a set of 1 184 334 autosomal SNPs was obtained. During the process of screening the optimal IBD length threshold, the result revealed that 0 cM, 1 cM, and 2 cM all demonstrated good applicability. However, to avoid the issue of a large amount of redundant information caused by setting a too low IBD length threshold, this study ultimately selected 2 cM as the optimal threshold. Compared with the average results of three chip panels, the high-density SNP locus set increased the total IBD length and the average IBD length across levels 1-9; the accuracy of the confidence interval for level 8 was 70.97%, which represented a 3.50% improvement; the average confidence interval accuracy for levels 1-8 was 91.39%, representing a 1.00% increase; and the false negative rates at levels 8 and 9 were reduced by 2.42% and 6.76%, respectively. The system efficacy of the high-density SNP locus set for kinship inference of first to eighth degree relationships reached 98.91%. Through random reduction of the high-density SNP locus set results, it is found that increasing the number of SNPs with the panel, the detection efficiency of IBD length showed a significant upward trend. At the same time, the overall trend in the accuracy of kinship relationship prediction as well as the confidence interval accuracy also indicated that both metrics steadily increased with the addition of more loci. ConclusionThe results show that the high-density SNPs panel significantly enhances the efficacy of distant kinship inference, accurately covering kinship degrees, with the average confidence interval accuracy for first to eighth degree relationships stably above 90%. The study finds that increasing the number of SNPs panel can improve the ability to predict distant kinship.

Spermatogenesis is a fundamental process that requires a tightly controlled epigenetic event in spermatogonial stem cells (SSCs). The mechanisms underlying the transition from SSCs to sperm are largely unknown. Most studies utilize gene knockout mice to explain the mechanisms. However, the production of genetically engineered mice is costly and time-consuming. In this study, we presented a convenient research strategy using an RNA interference (RNAi) and testicular transplantation approach. Histone H3 lysine 9 (H3K9) methylation was dynamically regulated during spermatogenesis. As Jumonji domain-containing protein 1A (JMJD1A) and Jumonji domain-containing protein 2C (JMJD2C) demethylases catalyze histone H3 lysine 9 dimethylation (H3K9me2), we firstly analyzed the expression profile of the two demethylases and then investigated their function. Using the convenient research strategy, we showed that normal spermatogenesis is disrupted due to the downregulated expression of both demethylases. These results suggest that this strategy might be a simple and alternative approach for analyzing spermatogenesis relative to the gene knockout mice strategy.
Spermatogenesis/physiology* ; Animals ; Male ; Mice ; Epigenesis, Genetic ; Jumonji Domain-Containing Histone Demethylases/metabolism* ; Histones/metabolism* ; RNA Interference ; Testis/metabolism* ; Methylation ; Mice, Knockout ; Histone Demethylases

AIM To investigate the effect of Fuzheng Hefu Zhiyang Formula(FZHFZY)on psoriasis-like skin lesions and immune regulation in mice.METHODS In the in vivo experiment,30 BALB/c mice were randomly divided into the blank group,the model group,the dexamethasone group(1.5 g/kg of compound dexamethasone acetate cream),and the low-dose(2.5 g/kg)and high-dose(5 g/kg)FZHFZY groups,with six mice in each group.The experiment groups were treated with respective FZHFZY and dexamethasone,and the other groups were given normal saline for 10 consecutive days,during which psoriatic skin lesions were induced with imiquimod cream for 7 consecutive days.The mice had their area and severity of psoriasis assessed by PASI score;their histological changes of skin lesions.observed with Hematoxylin-eosin(HE)staining;their F4/80 ratio of skin lesions observed with immunohistochemical(IHC)staining;their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected by Western blot;and their mRNA expressions of tumor necrosis factor-α(TNF-α),IL-17,IL-23 and IL-1β detected by RT-qPCR.In the in vitro research,the cultured RAW264.7 cells were divided into the blank group,the LPS group,and the FZHFZY groups(1 200,600,300,150 μg/mL).The cells had their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected with Western blot;and their mRNA expressions of IL-6,TNF-α,IL-23 and IL-8 detected by RT-qPCR.RESULTS The in vivo experiment showed that compared to the model group,the FZHFZY groups demonstrated decreased PASI score(P＜0.01);improved epidermal thickening and parakeratosis of skin lesions as revealed by HE staining result and increased expression of F4/80 in IHC staining sections;decreased protein expression ratios of p-P38/P38,p-ERK/Erk and p-P65/P65 in skin(P＜0.05,P＜0.01);and reduced mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β in the skin(P＜0.01).FZHFZY(0～2 400 μg/mL)showed no significant cytotoxicity towards RAW264.7 cells in vitro(P＞0.05).Compared to those of the LPS group,the cells exposed to FZHFZ at concentrations of 1 200 and 600 μg/mL demonstrated decreased protein expression ratios of p-P38/P38,p-ERK/Erk,and p-P65/P65(P＜0.05,P＜0.01);and significantly decreased mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β(P＜0.01).CONCLUSION FZHFZY alleviates imiquimod-induced psoriatic lesions in mice and suppresses inflammatory response in LPS-stimulated RAW264.7 cells by inhibiting P38/Erk/NF-κB signaling pathway.

OBJECTIVE Aimed at investigating whether reflux pharyngitis is an independent risk factor for the failure of type Ⅰ tympanoplasty for chronic otitis media.This is achieved by analyzing the relationship between the postoperative tympanic membrane healing in patients who underwent type Ⅰ tympanoplasty and pharyngolaryngeal reflux finding score(RFS).METHODS Patients who underwent type Ⅰ tympanoplasty in the Department of Otolaryngology Head and Neck Surgery,Nanshan People's Hospital,Shenzhen,China,from January 2023 to July 2024 were retrospectively included.All the patients received preoperative perfect nasal endoscopy,laryngoscopy,evaluation by the RFS questionnaire,preoperative otoscopy for tympanoplasty,pure tone hearing threshold,and temporal bone thin-layer CT examination.Postoperative otoscopic examination was performed to observe tympanic membrane healing and followed up for 3 months.The patients were divided into surgery success group and failure group based on the criterion of whether a complete tympanic membrane was formed by endoscopic examination within 3 months.The RFS scores of the two groups were statistically analyzed.RESULTS A total of 135 patients with an average age of 44.78 years(±12.22 years)took part in this study,with 60 males and 75 females included,and 68 left ears and 67 right ears involved.There were 120 patients in the surgery success group,and 15 patients in the failure group.Statistical analysis revealed that the RFS score of the patients in the tympanoplasty failure group was remarkably higher than that of the patients in the tympanoplasty success group.Moreover,there were significantly more cases with suspected reflux pharyngitis in the surgery failure group(P=0.007).Reflux-induced tympanic membrane lesion and reperforation mostly occurred in the central part of the tympanic membrane graft.CONCLUSION Reflux pharyngitis has been implicated with tympanoplasty failure,and thus may be a causative factor.Additionally,the RFS can be used to screen patients with chronic suppurative otitis media for suspected reflux pharyngitis.Findings from this work indicate that perioperative anti-reflux therapy,combined with dietary and lifestyle counselling for the patients who suffer from reflux pharyngitis and are about to undergo the tympanoplasty surgery may improve surgical success rate.

Objective:To explore the potential prognostic value of the HAMP(hepcidin)gene in gastric adenocarci-noma and its correlation with immune infiltration in gastric cancer.Methods:This study systematically analyzed the ex-pression characteristics of the HAMP gene using bioinformatics approaches based on mRNA data from 448 gastric ad-enocarcinoma tissues and non-tumor tissues in the TCGA database.Firstly,the cBioPortal platform was employed to analyze the genetic variation features of the HAMP gene,and the LinkedOmics database was used to evaluate the corre-lation between its methylation status and expression levels.Kaplan-Meier survival analysis with log-rank test was per-formed to assess the relationship between HAMP expression levels and patient prognosis.Furthermore,by integrating TIMER2.0 and TISIDB databases,we systematically evaluated the correlation between HAMP expression and immune-related genes as well as immune cell infiltration.Gene Set Enrichment Analysis(GSEA)was conducted to investigate HAMP-associated signaling pathway characteristics.Finally,STRING and Gepia databases were utilized to construct a protein-protein interaction network of HAMP and identify core interacting genes,comprehensively evaluating the role of HAMP in immune infiltration in gastric adenocarcinoma.Results:The expression level of HAMP was significantly higher in gastric adenocarcinoma tissues compared to normal tissues(P<0.01),and its elevated expression was strongly associ-ated with poor patient prognosis,manifested by significantly shorter overall survival(OS),progression-free survival(PFS),and post-progression survival(PPS)(all P<0.05).Genomic analysis revealed that HAMP mutations in gastric cancer were predominantly amplification-type,and its methylation level showed a positive correlation with mRNA expression(r=0.14,P<0.001).Immunological analysis demonstrated that high HAMP expression was significantly correlated with multiple key immune checkpoint molecules(PD-1:rho=0.274;PD-L1:rho=0.211;CTLA-4:rho=0.199,all P<0.001)and immune cell infiltration(dendritic cells:r=0.548;macrophages:r=0.414;neutrophils:r=0.374,all P<0.001).Pathway enrichment analy-sis indicated that the high HAMP expression group was significantly enriched in immune-related pathways including anti-gen presentation and NK cell-mediated cytotoxicity.Furthermore,protein-protein interaction network analysis identified core interacting genes such as TREM2 and TYROBP,suggesting that HAMP may participate in tumor immune regulation through specific molecular networks.Conclusion:HAMP is highly expressed in gastric cancer,and its high expression significantly reduces the survival time of gastric adenocarcinoma patients,demonstrating prognostic value.HAMP ex-pression is positively correlated with most immune-related genes in STAD and significantly associated with the abun-dance of multiple immune cell infiltration levels,serving as an independent prognostic factor related to immune infiltration.

Perception of recurrence risk in cardiovascular disease (CVD) patients plays a significant role in aspects such as their quality of life and treatment adherence. This paper reviews the theoretical foundations of recurrence risk perception, the conceptual origins and developmental process, measurement tools, influencing factors of recurrence risk perception in CVD patients, and research progress of recurrence risk perception in CVD management. The aim is to provide a basis for developing scientifically effective intervention measures for CVD patients in the future.

Objective:To explore the potential prognostic value of the HAMP(hepcidin)gene in gastric adenocarci-noma and its correlation with immune infiltration in gastric cancer.Methods:This study systematically analyzed the ex-pression characteristics of the HAMP gene using bioinformatics approaches based on mRNA data from 448 gastric ad-enocarcinoma tissues and non-tumor tissues in the TCGA database.Firstly,the cBioPortal platform was employed to analyze the genetic variation features of the HAMP gene,and the LinkedOmics database was used to evaluate the corre-lation between its methylation status and expression levels.Kaplan-Meier survival analysis with log-rank test was per-formed to assess the relationship between HAMP expression levels and patient prognosis.Furthermore,by integrating TIMER2.0 and TISIDB databases,we systematically evaluated the correlation between HAMP expression and immune-related genes as well as immune cell infiltration.Gene Set Enrichment Analysis(GSEA)was conducted to investigate HAMP-associated signaling pathway characteristics.Finally,STRING and Gepia databases were utilized to construct a protein-protein interaction network of HAMP and identify core interacting genes,comprehensively evaluating the role of HAMP in immune infiltration in gastric adenocarcinoma.Results:The expression level of HAMP was significantly higher in gastric adenocarcinoma tissues compared to normal tissues(P<0.01),and its elevated expression was strongly associ-ated with poor patient prognosis,manifested by significantly shorter overall survival(OS),progression-free survival(PFS),and post-progression survival(PPS)(all P<0.05).Genomic analysis revealed that HAMP mutations in gastric cancer were predominantly amplification-type,and its methylation level showed a positive correlation with mRNA expression(r=0.14,P<0.001).Immunological analysis demonstrated that high HAMP expression was significantly correlated with multiple key immune checkpoint molecules(PD-1:rho=0.274;PD-L1:rho=0.211;CTLA-4:rho=0.199,all P<0.001)and immune cell infiltration(dendritic cells:r=0.548;macrophages:r=0.414;neutrophils:r=0.374,all P<0.001).Pathway enrichment analy-sis indicated that the high HAMP expression group was significantly enriched in immune-related pathways including anti-gen presentation and NK cell-mediated cytotoxicity.Furthermore,protein-protein interaction network analysis identified core interacting genes such as TREM2 and TYROBP,suggesting that HAMP may participate in tumor immune regulation through specific molecular networks.Conclusion:HAMP is highly expressed in gastric cancer,and its high expression significantly reduces the survival time of gastric adenocarcinoma patients,demonstrating prognostic value.HAMP ex-pression is positively correlated with most immune-related genes in STAD and significantly associated with the abun-dance of multiple immune cell infiltration levels,serving as an independent prognostic factor related to immune infiltration.

Perception of recurrence risk in cardiovascular disease (CVD) patients plays a significant role in aspects such as their quality of life and treatment adherence. This paper reviews the theoretical foundations of recurrence risk perception, the conceptual origins and developmental process, measurement tools, influencing factors of recurrence risk perception in CVD patients, and research progress of recurrence risk perception in CVD management. The aim is to provide a basis for developing scientifically effective intervention measures for CVD patients in the future.

AIM To investigate the effect of Fuzheng Hefu Zhiyang Formula(FZHFZY)on psoriasis-like skin lesions and immune regulation in mice.METHODS In the in vivo experiment,30 BALB/c mice were randomly divided into the blank group,the model group,the dexamethasone group(1.5 g/kg of compound dexamethasone acetate cream),and the low-dose(2.5 g/kg)and high-dose(5 g/kg)FZHFZY groups,with six mice in each group.The experiment groups were treated with respective FZHFZY and dexamethasone,and the other groups were given normal saline for 10 consecutive days,during which psoriatic skin lesions were induced with imiquimod cream for 7 consecutive days.The mice had their area and severity of psoriasis assessed by PASI score;their histological changes of skin lesions.observed with Hematoxylin-eosin(HE)staining;their F4/80 ratio of skin lesions observed with immunohistochemical(IHC)staining;their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected by Western blot;and their mRNA expressions of tumor necrosis factor-α(TNF-α),IL-17,IL-23 and IL-1β detected by RT-qPCR.In the in vitro research,the cultured RAW264.7 cells were divided into the blank group,the LPS group,and the FZHFZY groups(1 200,600,300,150 μg/mL).The cells had their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected with Western blot;and their mRNA expressions of IL-6,TNF-α,IL-23 and IL-8 detected by RT-qPCR.RESULTS The in vivo experiment showed that compared to the model group,the FZHFZY groups demonstrated decreased PASI score(P＜0.01);improved epidermal thickening and parakeratosis of skin lesions as revealed by HE staining result and increased expression of F4/80 in IHC staining sections;decreased protein expression ratios of p-P38/P38,p-ERK/Erk and p-P65/P65 in skin(P＜0.05,P＜0.01);and reduced mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β in the skin(P＜0.01).FZHFZY(0～2 400 μg/mL)showed no significant cytotoxicity towards RAW264.7 cells in vitro(P＞0.05).Compared to those of the LPS group,the cells exposed to FZHFZ at concentrations of 1 200 and 600 μg/mL demonstrated decreased protein expression ratios of p-P38/P38,p-ERK/Erk,and p-P65/P65(P＜0.05,P＜0.01);and significantly decreased mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β(P＜0.01).CONCLUSION FZHFZY alleviates imiquimod-induced psoriatic lesions in mice and suppresses inflammatory response in LPS-stimulated RAW264.7 cells by inhibiting P38/Erk/NF-κB signaling pathway.

Objective:To investigate the effects and underlying mechanism of ionizing radiation on the adipogenic of mesenchymal stem cells(MSCs).Methods:Mouse MSCs were cultured in vitro and treated with 2 Gy and 6 Gy radiation with 60Co,and the radiation dose rate was 0.98 Gy/min.Bulk RNA-seq was performed on control and irradiated MSCs.The changes of adipogenic differentiation and oxidative stress pathways of MSC were revealed by bioinformatics analysis.Oil Red O staining was used to detect the adipogenic differentiation ability of MSCs in vitro,and real-time fluorescence quantitative PCR(qPCR)was used to detect the expression differences of key regulatory factors Cebpa,Lpl and Pparg after radiation treatment.At the same time,qPCR and Western blot were used to detect the effect of inhibition of Nrf2,a key factor of antioxidant stress pathway,on the expression of key regulatory factors of adipogenesis.Moreover,the species conservation of the irradiation response of human bone marrow MSCs and mouse MSC was determined by qPCR.Results:Bulk RNA-seq suggested that ionizing radiation promotes adipogenic differentiation of MSCs and up-regulation of oxidative stress-related genes and pathways.The results of Oil Red O staining and qPCR showed that ionizing radiation promoted the adipogenesis of MSCs,with high expression of Cebpa,Lpl and Pparg,as well as oxidative stress-related gene Nrf2.Nrf2 pathway inhibitors could further enhance the adipogenesis of MSCs in bone marrow after radiation.Notably,the similar regulation of oxidative pathways and enhanced adipogenesis post irradiation were observed in human bone marrow MSCs.In addition,irradiation exposure led to up-regulated mRNA expression of interleukin-6 and down-regulated mRNA expression of colony stimulating factor 2 in human bone marrow MSCs.Conclusion:Ionizing radiation promotes adipogenesis of MSCs in mice,and oxidative stress pathway participates in this effect,blocking Nrf2 further promotes the adipogenesis of MSCs.Additionally,irradiation activates oxidative pathways and promotes adipogenic differentiation of human bone marrow MSCs.