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.

Cowden syndrome is a rare autosomal dominant disorder caused by a variant of the PTEN gene,with clinical manifestations mainly of fetal overgrowth and organ tumors.The pregnant woman in this case was 32 years old with gravida 1 and para 0.Prenatal ultrasound in Obstetrics and Gynecology Hospital,Fudan University demonstrated fetal biological parameters abnormally increased(especially in biparietal diameter and circumference of the head)and hepatic tumor.Genetic testing suggested a PTEN gene variant[NM_000314.4:c.193T>G(p.Tyr65Asp)].After termination of pregnancy,autopsy pathology diagnosed fetal organs overgrowth and hepatic hemangioma.This article focuses on the prenatal ultrasound phenotype and genetic features of Cowden syndrome to increase clinicians'awareness of this rare disease.

The pregnant woman was 30 years old,G2P0.This singleton pregnancy at 22 weeks of gestation was screened for second-trimester ultrasound malformations,suggesting fetal aortic valve atresia,aortic stenosis with reverse blood flow,mitral valve atresia,and markedly enlarged left ventricle,which was considered for the diagnosis of hypoplastic left heart syndrome(HLHS).The pregnancy was terminated at our hospital and subsequently underwent genetic testing with results of heterozygous variants in the NOTCH1 gene,which can cause aortic valve disease type 1.The findings of the fetal autopsy were aortic valve atresia,mitral valve widening and thickening,and left ventricular enlargement with myocardial infarction.This report focuses on the ultrasound characteristics of HLHS with left ventricular enlargement and its hemodynamic changes in order to improve clinicians'understanding of the progressive changes in the disease phenotype of HLHS.

Cowden syndrome is a rare autosomal dominant disorder caused by a variant of the PTEN gene,with clinical manifestations mainly of fetal overgrowth and organ tumors.The pregnant woman in this case was 32 years old with gravida 1 and para 0.Prenatal ultrasound in Obstetrics and Gynecology Hospital,Fudan University demonstrated fetal biological parameters abnormally increased(especially in biparietal diameter and circumference of the head)and hepatic tumor.Genetic testing suggested a PTEN gene variant[NM_000314.4:c.193T>G(p.Tyr65Asp)].After termination of pregnancy,autopsy pathology diagnosed fetal organs overgrowth and hepatic hemangioma.This article focuses on the prenatal ultrasound phenotype and genetic features of Cowden syndrome to increase clinicians'awareness of this rare disease.

Objective:To explore whether LBP3 exerts anti-tumor effects by promoting production of short-chain fatty acids(SCFAs)by intestinal microbiota and regulating function of polymorphonuclear myeloid-derived suppressor cells(PMN-MDSC).Methods:A subcutaneous H22 liver cancer model was employed to assess anti-tumor activity of LBP3 and its regulatory effects on PMN-MDSC.Pseudo-sterile tumor-bearing mouse model was used to investigate role of intestinal microbiota in tumor suppression of LBP3.Fecal microbiota transplantation(FMT)was conducted to explore immune regulatory role of LBP3-modulated flora.Serum SCFAs levels in tumor-bearing mice were quantified using liquid chromatography-mass spectrometry,and effect of SCFAs butyrate on arginase 1(Arg-1)expression was evaluated in vitro.Results:Both low-dose(125 mg/kg)and high-dose(250 mg/kg)LBP3 signifi-cantly inhibited tumor growth in H22 tumor-bearing mice,also led to a marked reduction in proportion of PMN-MDSC in both spleen and tumor,a reduced proportion of Treg in lymphoid tissues,a decrease in Arg-1 level within tumor,infiltration of CD8+T cells into tumor was significantly enhanced.However,these effects of LBP3 were did not observed in pseudo-sterile mice,while the above changes could be reproduced after fecal supernatant transplantation in high-dose LBP3 treatment group,suggesting a crucial role for gut microbiota.Furthermore,co-expression of Ly6G and SCFA receptor GPR43 in tumor was also observed.LBP3 treatment resulted in increased levels of SCFAs,particularly butyrate,in both blood and tumor tissues.In vitro,butyrate was shown to inhibit Arg-1 expression in MSC-2 cells,further supporting hypothesis that SCFAs mediate immune-modulatory effects of LBP3.Conclusion:LBP3 exerts its anti-tumor effects by promoting SCFA production,which subsequently inhibits function of PMN-MDSC.This highlights LBP3's potential as an immunomodulatory agent in cancer therapy.

Objective To identify key proteins associated with sudden cardiac death(SCD)caused by lethal arrhythmia and to explore their potential molecular mechanisms through integrated proteomic analysis,data mining,and bioinformatics.Methods A lethal arrhythmia model was established in 8-week-old male Sprague-Dawley rats,which were divided into an arrhythmia group and a control group.Proteomic techniques were applied to identify and quantify proteins in left ventricular myocardial tissue,and differentially expressed proteins related to arrhythmia were screened.Key proteins were further identified through comparison with target proteins in databases combined with joint analyses.Bioinformatics methods were then used to investigate potential molecular mechanisms.Results A total of 356 differentially expressed proteins were identified,including 189 upregulated and 167 downregulated.Association analysis with target gene proteins identified 71 key proteins,and a protein-protein interaction network was constructed.GO enrichment and KEGG pathway analyses indicated that these key proteins were primarily involved in ion channel dysfunction,enhanced oxidative stress,and autonomic nervous system imbalance.Conclusion This study,through the integration of proteomics,data mining,and bioinformatics,revealed critical molecular mechanisms underlying SCD associated with lethal arrhythmia.These findings provide new perspectives and potential biomarkers for forensic identification and research on the mechanisms of death.

BACKGROUND:Skeletal muscle insulin resistance is the key pathological link of type 2 diabetes.The traditional Chinese medicine compound Roujishuncuiyin can effectively improve skeletal muscle insulin resistance,but its mechanism has not been clarified.OBJECTIVE:To explore the mechanism of Roujishuncuiyin on skeletal muscle insulin resistance in type 2 diabetes mice.METHODS:Forty db/db mice with type 2 diabetes mellitus were randomized into a model group,a low-dose Roujishuncuiyin group,a high-dose Roujishuncuiyin group,and a positive drug group,with 10 mice in each group.The latter three administration groups were given 157.5 mg/g and 630 mg/g Roujishuncuiyin and 200 mg/g metformin hydrochloride aqueous solution by gavage once a day,respectively.In addition,10 db/dm mice were selected as the blank control group.Mice in the model and blank control groups were given the same dose of 0.9％NaCl solution by gavage.After 12 weeks of intervention,fasting blood glucose was measured in each group of mice,and oral glucose tolerance test was performed to calculate the area under the blood glucose curve.ELISA was used to detect serum insulin level and calculate the resistance index.Mitochondrial structure of skeletal muscle tissue was observed under transmission electron microscopy.Western blot was used to detect the expression levels and phosphorylation levels of protein kinase B(AKT)and glycogen synthase kinase 3β(GSK-3β)proteins in skeletal muscle.RESULTS AND CONCLUSION:(1)Compared with the blank control group,fasting blood glucose,fasting insulin and insulin resistance index were significantly higher in the model group(P＜0.05),the area under the curve of the oral glucose tolerance test was significantly increased(P＜0.05),the expression of p-AKT and p-GSK3β proteins in tibialis anterior muscle was significantly decreased(P＜0.05),and there was a large amount of mitochondrial damage in tibialis anterior muscle and a large number of lipid droplets in the interstitium.(2)Compared with the model group,fasting blood glucose,fasting insulin,and insulin resistance index were significantly reduced in the low-and high-dose Roujishuncuiyin groups and the positive control group(P＜0.05),the area under the curve of the oral glucose tolerance test was reduced(P＜0.05),the expression of p-AKT and p-GSK3β proteins in the tibialis anterior muscle was significantly elevated(P＜0.05),and mitochondrial damage in the tibialis anterior muscle was significantly ameliorated,with decreased lipid droplets in the interstitium.(3)The above indexes were better in the high-dose Roujishuncuiyin group than the low-dose Roujishuncuiyin group(P＜0.05),while there was no significant difference between the high-dose Roujishuncuiyin group and positive control group(P＞0.05).To conclude,by upregulating the protein levels of p-AKT and p-GSK3β in skeletal muscle tissue,the traditional Chinese medicine compound Roujishuncuiyin can improve structural disorders and mitochondrial morphology in skeletal muscle tissue,reduce insulin resistance in the skeletal muscle and regulate glucose homeostasis in the body.

Background and Aims:Long noncoding RNA(lncRNA)nuclear-enriched abundant transcript 1(NEAT1)is an oncogenic lncRNA that promotes the progression of various cancers through the competing endogenous RNA(ceRNA)mechanism.Using the TargetScan database,we previously identified binding sites between NEAT1 and microRNA-124-3p(miR-124-3p),as well as between miR-124-3p and catenin beta-1(CTNNB1).Therefore,this study was conducted to investigate the expression of NEAT1,miR-124-3p,and CTNNB1 in pancreatic cancer and their interactions affecting pancreatic cancer cell functions.Methods:A dual-luciferase reporter assay was used to validate the relationships among NEAT1,miR-124-3p,and CTNNB1.The expression levels of NEAT1 and miR-124-3p,as well as CTNNB1 protein expression,were detected in pancreatic cancer tissues and adjacent normal tissues,as well as in pancreatic cancer PANC-1 cells and normal pancreatic epithelial H6C7 cells.PANC-1 cells were transfected with NEAT1 siRNA alone or co-transfected with a miR-124-3p inhibitor.After transfection,changes in PANC-1 cell biological functions,epithelial-mesenchymal transition related protein expression,and tumor growth ability in mice were assessed.Results:The dual-luciferase reporter assay confirmed the targeting relationships between NEAT1 and miR-124-3p,as well as between miR-124-3p and CTNNB1.NEAT1 and CTNNB1 expression levels were significantly upregulated,while miR-124-3p expression was downregulated in pancreatic cancer tissues(vs.adjacent tissues)and in PANC-1 cells(vs.H6C7 cells)(all P<0.05).NEAT1 siRNA transfection led to decreased NEAT1 and CTNNB1 expression and increased miR-124-3p expression in PANC-1 cells.However,co-transfection with a miR-124-3p inhibitor suppressed the expression changes in miR-124-3p and CTNNB1(all P<0.05).NEAT1 siRNA transfection significantly reduced PANC-1 cell proliferation,migration,and invasion,while promoting apoptosis.Additionally,E-cadherin protein expression was upregulated,whereas N-cadherin and vimentin protein expression were downregulated.Tumor growth in mice was also significantly inhibited(all P<0.05).These changes were attenuated upon co-transfection with the miR-124-3p inhibitor(all P<0.05).Conclusion:NEAT1 may act as a ceRNA by competitively binding to miR-124-3p,thereby attenuating miR-124-3p-mediated inhibition of CTNNB1.This leads to CTNNB1 upregulation,ultimately promoting the malignant biological behavior of pancreatic cancer cells.

Background:Ribosomal protein L22-like 1(RPL22L1)exerts regulatory effects on various malignant tumors such as lung cancer,prostate cancer,and cervical cancer.However,its role in colorectal cancer(CRC)remains unclear.Aims:To investigate the expression of RPL22L1 in CRC and its role in patients' prognosis and glucose metabolism of tumor cells.Methods:A total of 142 newly diagnosed CRC patients admitted to the Taizhou First People's Hospital from February 2022 to June 2024 were enrolled.The expression levels of RPL22L1 mRNA and protein were detected by quantitative real-time PCR and immunohistochemistry,respectively.The correlation between RPL22L1 expression and clinicopathological characteristics was analyzed.Kaplan-Meier survival analysis was used to evaluate the impact of RPL22L1 expression on the prognosis of CRC patients.RPL22L1 siRNA was transfected into SW480 cells to establish a low-expression cell model.Cell proliferation was assessed by CCK-8 assay,cell migration by Transwell chamber assay,and apoptosis by flow cytometry.Gene set enrichment analysis(GSEA)was performed to evaluate the effect of RPL22L1 on glucose metabolism of tumor cells.Results:The expression levels of RPL22L1 mRNA and protein were significantly higher in CRC tissues than in adjacent normal tissues(all P<0.05).The expression level of RPL22L1 mRNA was correlated with the TNM stage and carcinoembryonic antigen level of CRC(all P<0.05).Kaplan-Meier analysis showed that the cumulative survival rate of high RPL22L1 mRNA expression group was significantly lower than that of low-expression group(P=0.027).The expression level of RPL22L1 mRNA was significantly higher in SW480 cells than in normal intestinal epithelial cells(P<0.001).After inhibiting RPL22L1 expression,the proliferation and migration capacities of SW480 cells were significantly decreased(all P<0.05),the apoptosis rate was significantly increased(P=0.005),and the lactate level and relative glucose uptake level were significantly reduced(all P<0.05).GSEA indicated that RPL22L1 gene was associated with glycolysis/gluconeo-genesis(P=0.02).Conclusions:RPL22L1 is highly expressed in CRC and is associated with poor prognosis of patients,suggesting its potential as a molecular target for CRC therapy.Furthermore,RPL22L1 may promote the tumorigenesis and progression of CRC by modulating glucose metabolism.

Objective:To analyze the revision strategies for failed atlantoaxial dislocation (AAD) surgery.Methods:A retrospective analysis was conducted on 145 patients who underwent revision surgery for AAD at the General Hospital of Southern Theatre Command of PLA between September 2009 and December 2023. The cohort included 74 males and 71 females, with a mean age of 43±16 years (range, 6-72 years). The initial surgical approaches were: anterior 31 cases, posterior 114 cases. Based on imaging assessments of immediate postoperative reduction and fusion status prior to revision, the cases of failure were classified into reduction-nonfusion type (22 cases), nonreduction-fusion type (31 cases), and nonreduction-nonfusion type (92 cases). Among the nonreduction-nonfusion cases, 39 had initial surgery with internal fixation for reduction, while 53 had initial surgery with simple decompression (posterior arch resection, foramen magnum decompression) without reduction. In the nonreduction-fusion cases, 8 cases had spot fusion and 23 had extensive fusion. Japanese Orthopaedic Association (JOA) scores were compared before and after revision, and complication rates were observed.Results:All patients successfully underwent surgery. The revision approaches included: anterior (anterior fixation and fusion 52 cases, anterior implant removal combined anterior fixation and fusion 4 cases, transoral odontoidectomies 16 cases, anterior implant removal combined transoral odontoidectomy 2 cases), posterior (posterior fixation and fusion 2 cases, posterior implant removal combined posterior fixation and fusion 22 cases), and combined anterior-posterior (posterior implant removal combined anterior fixation and fusion 18 cases, anterior implant removal combined posterior fixation and fusion 25 cases, posterior implant removal combined transoral odontoidectomy 5 cases). Operative time was 254.20±107.63 min (range, 90-660 min), and blood loss was 218.83±172.17 ml (range, 20-800 ml). Except for 3 patients who died due to postoperative complications, all patients were followed up for a duration of 12±11 months (range, 3-60 months). Six patients who failed to achieve bony fusion after the initial revision surgery underwent a second revision due to poor reduction (1 case), infection (1 case), suboptimal implant position (3 cases), and graft nonunion (1 case). All three patients with bony fusion after the initial revision surgery underwent a second revision due to poor reduction. Following the second revision surgery, none of the 9 patients exhibited graft nonunion or spinal cord compression. The 136 successful initial revision cases had a final follow-up JOA score of 14.75±2.00, significantly higher than the preoperative score of 11.93±2.92 ( t=-18.869, P<0.001). Conclusions:Revision surgery for AAD should take into account the immediate postoperative reduction status and fusion status prior to revision. An appropriate revision strategy should be selected to achieve satisfactory reduction and bony fusion.