Objective:To explore the clinical characteristics of patients with porto-sinusoidal vascular disease (PSVD) complicated with esophageal and gastric varices, and to evaluate the effect of portal vein pressure (PVP) on esophageal and gastric varices bleeding and rebleeding after endoscopic treatment.Methods:Patients who were hospitalized in the Department of Gastroenterology of the Zhongshan Hospital, Fudan University due to portal hypertension from July 2022 to October 2024, underwent liver biopsy for diagnosis of PSVD, and received direct PVP measurement were included. Their clinical manifestations, liver histopathological characteristics were analyzed, and the prognosis was followed up.Results:A total of 29 patients were included, and 19 patients had experienced rupture and bleeding of esophageal and gastric varices. Compared with the non-bleeding group, the hemoglobin level of patients in the bleeding group was lower and the international normalized ratio was higher (all P<0.05). There was no statistically significant difference between the two groups in other laboratory examination indicators, complications of portal hypertension, combined diseases, etc. (all P>0.05). The pathology of liver biopsy suggests that dilation of the hepatic sinuses and abnormalities of the central vein are common pathological changes. The direct PVP of patients in the bleeding group was significantly higher than that in the non-bleeding group [28.0(24.5-31.0)mmHg vs 18.5(10.5-23.8)mmHg, P=0.011]. However, there was no statistically significant difference in the measured values of liver wedge pressure, free pressure and hepatic venous pressure gradient (HVPG) between the two groups of patients (all P>0.05). Correlation analysis revealed that there was no significant correlation between HVPG and PVP ( R2=0.129 9, P=0.076 7). Grouped according to the median PVP value of 25 mmHg, the risk of esophageal and gastric variceal rupture and bleeding in the high PVP group (≥25 mmHg) was significantly higher than that in the low PVP group (<25 mmHg) (14/16 vs 5/13, P=0.016). The risk of rebleeding after endoscopic treatment in patients with high PVP (4/13) was higher than that in patients with low PVP (0/4). Conclusions:Patients with porto-sinusoidal vascular disease complicated with portal hypertension are often accompanied by rupture and bleeding of esophageal and gastric varices. HVPG cannot accurately reflect the portal vein pressure. The risk of rupture and bleeding of esophageal and gastric varices and rebleeding in patients with elevated portal vein pressure is significantly increased.

BACKGROUND:Programmed cell death protein 1 belongs to the immunoglobulin gene superfamily and can regulate the differentiation of osteoblasts and affect bone homeostasis.However,there are few studies on the regulatory role and mechanism of programmed cell death protein 1 in diabetic osteoporosis.OBJECTIVE:To investigate the regulatory role and mechanism of programmed cell death protein 1 on osteogenic differentiation of rat bone marrow mesenchymal stem cells under high-glucose environment.METHODS:(1)Animal experiment:A total of 12 Sprageu-Dawley rats were randomized into a control group(n=6)and a model group(n=6).The control group was fed routinely,whereas the model group was injected intraperitoneally with streptozotocin to establish a model of type 1 diabetes mellitus,and the high-fat feed was fed for 8 weeks to establish a model of type 1 diabetic osteoporosis.After 8 weeks of feeding,the femurs of rats in the two groups were taken and subjected to hematoxylin-eosin staining and micro-CT assay.The mRNA expression of programmed cell death protein 1 and programmed death ligand 1 was detected.(2)Cell experiment:Passage 3 rat bone marrow mesenchymal stem cells were randomly divided into four groups:normal control group,high-glucose model group cultured in low glucose medium,programmed cell death protein 1-silenced group transfected with programmed cell death protein 1 siRNA,and programmed cell death protein 1-silenced null group transfected with siRNA-NC.After 48 hours of transfection,the normal control group was cultured in a new low-glucose medium,and the other three groups were cultured in a high-glucose medium for another 48 hours of culture followed by osteogenic induction.After 21 days of osteogenic induction,alizarin red staining,and qRT-PCR(programmed cell death protein 1 and RUNX2 mRNA expression)and western blot(β-catenin,GSK-3β,p-GSK-3β and Axin2 protein expression)were performed.RESULTS AND CONCLUSION:In the animal experiment,hematoxylin-eosin staining and micro-CT assay showed successful modeling of type 1 diabetic osteoporosis in the model group.qRT-PCR assay showed that the mRNA expression of programmed cell death protein 1 and programmed cell death ligand 1 was higher in the model group than the control group(P<0.05).In the cell experiment,the results of alizarin red staining showed that the ability of mineralized nodule formation was lower in the high-glucose model group and the programmed cell death protein 1-silenced null group than in the control group and the programmed cell death protein 1-silenced group.Compared with the normal control group,the programmed cell death protein 1 mRNA expression and GSK3β and Axin2 protein expression were elevated in the high-glucose model group and the programmed cell death protein 1-silenced null group(P<0.05),and the RUNX2 mRNA expression and p-GSK3β and β-catenin protein expression were decreased(P<0.05).Compared with the high-glucose model group and the programmed cell death protein 1-silenced null group,programmed cell death protein 1 mRNA expression and GSK3β and Axin2 protein expression were decreased in the programmed cell death protein 1-silenced group(P<0.05),and RUNX2 mRNA expression and p-GSK3β and β-catenin protein expression were elevated(P<0.05).To conclude,programmed cell death protein 1 silencing can activate the Wnt/β-catenin and improve the osteogenic differentiation of rat bone marrow mesenchymal stem cells under high-glucose conditions.

Purpose To investigate the correlation between the methylation level at CpG sites of CXCL17 and the clinicopathological parameters of papillary thyroid carcinoma(PTC).Methods samples from 186 cases of PTC and 191 cases of benign thyroid nodule(BTN)were collected.Methylation levels of CXCL17 were semi-quantitatively as-sessed using mass spectrometry.Logistic regression analysis,which adjusted for age,gender and related hormones,was conducted to evaluate the correlation between CXCL17 methylation and PTC,and calculate the odds ratios(ORs)and 95％confidence intervals(CIs).Results Hypomethylation level of CXCL17_CpG_1.2 was significantly associat-ed with PTC(OR=1.36,95％CI:1.16-1.60,P＜0.001)and early stage of PTC patients(Stage Ⅰ,OR=1.41,95％CI:1.19-1.67,P＜0.001).Gender-based hierarchical management analysis showed that decreased methyla-tion level of CXCL17_CpG_1.2 was significantly associated with female PTC patients(OR=1.39,95％CI:1.15-1.67,P＜0.001).In subgroups stratified by age(＜50 and≥50 years old),hypomethylation at CXCL17_CpG_1.2 was significantly associated with PTC,with a stronger association in the younger subgroup(＜50 years old:OR=1.42,95％CI:1.14-1.77,P＜0.01;≥ 50 years old:OR=1.30,95％CI:1.03-1.64,P＜0.05).Conclusion There was a significant difference in CXCL17 methylation levels between benign and malignant thyroid tumors.It was showed that hypomethylation of CXCL17 is closely associated with PTC,particularly in young women patient.Thus,CXCL17 methylation may serve as a biomarker for accurate differential diagnosis of thyroid nodule.

Objective:To explore the clinical characteristics of patients with porto-sinusoidal vascular disease (PSVD) complicated with esophageal and gastric varices, and to evaluate the effect of portal vein pressure (PVP) on esophageal and gastric varices bleeding and rebleeding after endoscopic treatment.Methods:Patients who were hospitalized in the Department of Gastroenterology of the Zhongshan Hospital, Fudan University due to portal hypertension from July 2022 to October 2024, underwent liver biopsy for diagnosis of PSVD, and received direct PVP measurement were included. Their clinical manifestations, liver histopathological characteristics were analyzed, and the prognosis was followed up.Results:A total of 29 patients were included, and 19 patients had experienced rupture and bleeding of esophageal and gastric varices. Compared with the non-bleeding group, the hemoglobin level of patients in the bleeding group was lower and the international normalized ratio was higher (all P<0.05). There was no statistically significant difference between the two groups in other laboratory examination indicators, complications of portal hypertension, combined diseases, etc. (all P>0.05). The pathology of liver biopsy suggests that dilation of the hepatic sinuses and abnormalities of the central vein are common pathological changes. The direct PVP of patients in the bleeding group was significantly higher than that in the non-bleeding group [28.0(24.5-31.0)mmHg vs 18.5(10.5-23.8)mmHg, P=0.011]. However, there was no statistically significant difference in the measured values of liver wedge pressure, free pressure and hepatic venous pressure gradient (HVPG) between the two groups of patients (all P>0.05). Correlation analysis revealed that there was no significant correlation between HVPG and PVP ( R2=0.129 9, P=0.076 7). Grouped according to the median PVP value of 25 mmHg, the risk of esophageal and gastric variceal rupture and bleeding in the high PVP group (≥25 mmHg) was significantly higher than that in the low PVP group (<25 mmHg) (14/16 vs 5/13, P=0.016). The risk of rebleeding after endoscopic treatment in patients with high PVP (4/13) was higher than that in patients with low PVP (0/4). Conclusions:Patients with porto-sinusoidal vascular disease complicated with portal hypertension are often accompanied by rupture and bleeding of esophageal and gastric varices. HVPG cannot accurately reflect the portal vein pressure. The risk of rupture and bleeding of esophageal and gastric varices and rebleeding in patients with elevated portal vein pressure is significantly increased.

Purpose To investigate the correlation between the methylation level at CpG sites of CXCL17 and the clinicopathological parameters of papillary thyroid carcinoma(PTC).Methods samples from 186 cases of PTC and 191 cases of benign thyroid nodule(BTN)were collected.Methylation levels of CXCL17 were semi-quantitatively as-sessed using mass spectrometry.Logistic regression analysis,which adjusted for age,gender and related hormones,was conducted to evaluate the correlation between CXCL17 methylation and PTC,and calculate the odds ratios(ORs)and 95％confidence intervals(CIs).Results Hypomethylation level of CXCL17_CpG_1.2 was significantly associat-ed with PTC(OR=1.36,95％CI:1.16-1.60,P＜0.001)and early stage of PTC patients(Stage Ⅰ,OR=1.41,95％CI:1.19-1.67,P＜0.001).Gender-based hierarchical management analysis showed that decreased methyla-tion level of CXCL17_CpG_1.2 was significantly associated with female PTC patients(OR=1.39,95％CI:1.15-1.67,P＜0.001).In subgroups stratified by age(＜50 and≥50 years old),hypomethylation at CXCL17_CpG_1.2 was significantly associated with PTC,with a stronger association in the younger subgroup(＜50 years old:OR=1.42,95％CI:1.14-1.77,P＜0.01;≥ 50 years old:OR=1.30,95％CI:1.03-1.64,P＜0.05).Conclusion There was a significant difference in CXCL17 methylation levels between benign and malignant thyroid tumors.It was showed that hypomethylation of CXCL17 is closely associated with PTC,particularly in young women patient.Thus,CXCL17 methylation may serve as a biomarker for accurate differential diagnosis of thyroid nodule.

BACKGROUND:Programmed cell death protein 1 belongs to the immunoglobulin gene superfamily and can regulate the differentiation of osteoblasts and affect bone homeostasis.However,there are few studies on the regulatory role and mechanism of programmed cell death protein 1 in diabetic osteoporosis.OBJECTIVE:To investigate the regulatory role and mechanism of programmed cell death protein 1 on osteogenic differentiation of rat bone marrow mesenchymal stem cells under high-glucose environment.METHODS:(1)Animal experiment:A total of 12 Sprageu-Dawley rats were randomized into a control group(n=6)and a model group(n=6).The control group was fed routinely,whereas the model group was injected intraperitoneally with streptozotocin to establish a model of type 1 diabetes mellitus,and the high-fat feed was fed for 8 weeks to establish a model of type 1 diabetic osteoporosis.After 8 weeks of feeding,the femurs of rats in the two groups were taken and subjected to hematoxylin-eosin staining and micro-CT assay.The mRNA expression of programmed cell death protein 1 and programmed death ligand 1 was detected.(2)Cell experiment:Passage 3 rat bone marrow mesenchymal stem cells were randomly divided into four groups:normal control group,high-glucose model group cultured in low glucose medium,programmed cell death protein 1-silenced group transfected with programmed cell death protein 1 siRNA,and programmed cell death protein 1-silenced null group transfected with siRNA-NC.After 48 hours of transfection,the normal control group was cultured in a new low-glucose medium,and the other three groups were cultured in a high-glucose medium for another 48 hours of culture followed by osteogenic induction.After 21 days of osteogenic induction,alizarin red staining,and qRT-PCR(programmed cell death protein 1 and RUNX2 mRNA expression)and western blot(β-catenin,GSK-3β,p-GSK-3β and Axin2 protein expression)were performed.RESULTS AND CONCLUSION:In the animal experiment,hematoxylin-eosin staining and micro-CT assay showed successful modeling of type 1 diabetic osteoporosis in the model group.qRT-PCR assay showed that the mRNA expression of programmed cell death protein 1 and programmed cell death ligand 1 was higher in the model group than the control group(P<0.05).In the cell experiment,the results of alizarin red staining showed that the ability of mineralized nodule formation was lower in the high-glucose model group and the programmed cell death protein 1-silenced null group than in the control group and the programmed cell death protein 1-silenced group.Compared with the normal control group,the programmed cell death protein 1 mRNA expression and GSK3β and Axin2 protein expression were elevated in the high-glucose model group and the programmed cell death protein 1-silenced null group(P<0.05),and the RUNX2 mRNA expression and p-GSK3β and β-catenin protein expression were decreased(P<0.05).Compared with the high-glucose model group and the programmed cell death protein 1-silenced null group,programmed cell death protein 1 mRNA expression and GSK3β and Axin2 protein expression were decreased in the programmed cell death protein 1-silenced group(P<0.05),and RUNX2 mRNA expression and p-GSK3β and β-catenin protein expression were elevated(P<0.05).To conclude,programmed cell death protein 1 silencing can activate the Wnt/β-catenin and improve the osteogenic differentiation of rat bone marrow mesenchymal stem cells under high-glucose conditions.

Objective Five chloroplast(cp)genomes from members of genus Polygonatum were assembled by hybrid assembly technique,and their intraspecic and interspecific differences were analyzed by comparative genomic method.Codon usage patterns and influencing factors were determined,and the cp genome data were applied to understand the phylogenomic relationships in the entire genus Polygonatum along with the available data.Methods In this study,the chloroplast genomes of 5 species of genus Polygonatum were assembled using Unicycler software.Sequence alignment,collinearity analysis,boundary analysis and other methods were used to evaluate the interspecific differences of these five species.Nucleotide polymorphism analysis was used to discover the high-variation sites of the five species and their related species,predict the distribution of different long repeat sequences and SSRs,and then analyze the use bias of Polygonatum code.Finally,phylogenetic tree was constructed with the coding sequences of other 47 genus Polygonatum and their closely related species to explore their phylogenetic relationships in this study.Results ①Chloroplast genomes of 155 408-155 623 bp were assembled from five species of Polygonatum.A total of 132-133 genes were annotated,and 369 long repeats and 1553 simple repeats were detected.②The contraction and expansion of chloroplast genomes in 8 species were not obvious at the IRs boundary,and the size and distribution of individual genes at the LSC-IRs-SSC boundary,such as ndhF gene and ycf1 gene,were slightly different.No interspecific or intraspecific rearrangement was observed in 8 species.③ The high-variation regions of the 8 chloroplast genomes are mainly located in two single-copy regions,the duplicate copy region is relatively conserved,and the coding region is more conserved than the non-coding region.High nucleotide polymorphic loci rps16-trnQ,trnS-trnG,trnTUGU-trnL,ndhF-rpl32 and rpl32-trnL are located in the single copy region and most of them are gene spacer regions.④ The codon preference results showed that the codon preference of the five species was similar and mainly affected by selection pressure,and the third base of the codon played A dominant role and mainly ended in A/U.RSCU clustering heat map shows that PK and PZ,PCB and PS have close relationship.⑤ Phylogenetic trees divided 52 species into five branches:Ⅰ,Ⅱ,Ⅲ,ⅣandⅤ.PS,PK,PCB,PCZ and PZ were divided into ⅣandⅤbranches,among which PK and PZ were most closely related,while PCZ was more distant than the other four,was divided into the Ⅴbranch alone.Conclusion This study provided a reference for the phylogenetic research and molecular marker development of the medicinal plants of the Polygonum genus.

Objective:To establish a cutoff level of anti-Müllerian hormone(AMH)which could help with the di-agnosis of polycystic ovary syndrome(PCOS)in adults,and to analyze the risk factors of metabolic syndrome(MS).Methods:A retrospectively analyzed 426 PCOS patients(PCOS group)and 205 healthy controls aged 20-39 years from the Health Checkup Center of the Gynecological Endocrine Center,Hunan Maternal and Child Health Hospital from January 2021 to December 2023.AMH diagnostic validity was estimated by receiver operating characteristic(ROC)curves.Patients were subgrouped into PCOS combined metabolic syndrome group(MS-PCOS)and the uncomplicated MS group(UMS-PCOS)according to metabolic status.Multivariate Logistic regression analysis was performed to determine the risk factors for MS in PCOS patients.Results:The serum AMH level was higher in PCOS group than that in the control group(8.42±3.71 ng/ml vs.2.99±0.94 ng/ml,P＜0.001).AMH cutoff for the diagnosis of PCOS was determined as≥4.87 ng/ml on ROC analysis,and the area under the curve is 0.981 with 92.7％sensitivity and 94.6％specificity.The prevalence of MS was 18.3％(78 ca-ses)in PCOS group.Subgroup analysis showed that MS-PCOS patients had higher waist circumference,BMI,fasting glucose,dyslipidemia,hypertension(BP＞130/85 mmHg),and hormone related index androgen level,but lower AMH vs.UMS-PCOS.Multivariate Logistic regression analysis identified insulin resistance(OR 39.17,95％CI 9.33-164.48),BMI ≥24 kg/m2(OR 3.72,95％CI 1.86-7.45),and hyperandrogenism(OR 2.56,95％CI 1.34-4.89)as independent risk factors of MS.AMH was negatively associated with MS,a single-unit increase in AMH was associated with an 17％decrease in odds of MS(OR 0.83,95％CI 0.73-0.95,P=0.006).Conclusions:Serum AMH levels were significantly higher in adult PCOS patients,with an optimal diagnostic threshold of 4.87 ng/ml.Hyperandrogenism and low AMH levels may predict a higher risk of MS,in addition to metabolism-related factors.

Objective To identify the potential pathological mechanisms of tuberculous spondylitis(TS).Methods Spinal specimens were collected from 13 TS patients and 13 controls who received treatment at a hospital from March 2021 to March 2023.Specimens were randomly selected from 3 TS patients and 3 controls to perform high-throughput lncRNAs and mRNAs sequencing with Illumina NovaSeq 6000.Differentially expressed lncRNAs(DELncRs)and mRNAs(DEmRs)in TS specimens were identified and analyzed through differential expression,and enrichment analysis was performed.The top 20 DEmRs with high connectivity were identified through protein-protein interaction(PPI)network.Regulatory network of DElncRs and DEmRs was built.Finally,gene expression of the remaining specimens was analyzed using qRT-PCR detection.Results A total of 1 243 DEmRs and 262 DElncRs were identified.Enrichment analysis revealed that muscle contraction,muscle system processes,muscle structural development,PI3K Akt signaling pathway,calcium signaling pathway,and cAMP signaling pathway were activated in TS,while responses to cytokines,cytokine-mediated signaling pathways,regulation of immune system processes,cytokine-cytokine receptor interactions,human T-cell leukemia virus type 1 infection,and phago-somes were inhibited in TS.Three sub-networks were identified in PPI,among which MYL1,TTN,LOC102723407,HLA-A,interleukin(IL)-6,and IL-1β had the highest connectivity and were identified as key DEmRs.MYL1,TTN,and IL-6 were regulated by DElncRs.qRT-PCR validated the differential expression of key DEmRs in TS.Conclusion DEmRs are regulated by lncRNAs and participate in the pathological process of TS,and the immune responses are inhibited in diseases condition.This study reveals key molecules and signaling pathways in TS,providing new insights into the pathological mechanisms of TS,and suggest scientific basis for developing new therapeutic targets.

Objective To establish the reference interval of urine Alzheimer-associated neuronal thread protein(AD7c-NTP)for healthy adults in Mianyang area using the indirect method.Methods The detection results of urine AD7c-NTP from 5 093 healthy in-dividuals were collected from the information management database of Medical Laboratory Department of Sichuan Science City Hospital from March 2017 to March 2022.Skewness-kurtosis and Kolmogorov-Smirnov tests were used to determine whether the data followed a normal distribution.After removing outliers using the Box Plots method,the enrolled subjects were grouped by gender and age.The Mann-Whitney U or Kruska-Wallis H tests were used to analyze the between-group differences of urine AD7c-NTP in healthy individu-als with different genders and ages.The adjacent age groups without statistically significant difference(P＞0.05)were combined,and the indirect method(non-parametric test method)was used to calculate the reference intervals for different gender and age groups.Results Skewness-kurtosis and Kolmogorov-Smirnov tests showed that the data followed a non-normal distribution.After removing 293 outliers using the Box Plots method,a total of 4 800 subjects,including 3 199 males and 1 601 females,were enrolled.The enrolled subjects were grouped by gender and age,and the non-parametric test method were used to establish the reference intervals of urine AD7c-NTP in healthy populations with different genders.The Mann-Whitney U test confirmed that urine AD7c-NTP levels existed gen-der differences(Z=14.09,P＜0.01),and the reference intervals for males and females were≤1.10 ng/mL and≤1.40 ng/mL,re-spectively.There were also statistical differences in urine AD7c-NTP levels among different age groups of the same gender.After combi-ning adjacent age groups without statistically significant difference(P＞0.05),the reference intervals of urine AD7c-NTP in healthy populations with different genders and ages were established by the non-parametric test method,which were≤1.00 ng/mL for male 20-39 years old group,≤1.10 ng/mL for male 40-79 years old group,≤1.60 ng/mL for male≥80 years old group,≤1.30 ng/mL for female 20-69 years old group,and≤1.60 ng/mL for female≥70 years old group,respectively.The established reference intervals of urine AD7c-NTP were further verified by healthy individuals,and the results met the standards.Conclusion The reference intervals of urine AD7c-NTP in healthy populations with different genders and ages in Mianyang area are established successfully using the indi-rect method,which may help to predict the risk of Alzheimer's disease in clinical practice and provide support for the diagnosis and treatment of the disease.