Objective To establish an individualized nomogram model and evaluate its efficacy to provide a possible evaluation basis for the prognosis of lower third and abdominal part of oesophageal adenocarcinoma (EAC). Methods Lower third and abdominal part of EAC patients from 2010 to 2015 were chosen from the SEER Research Plus Database (17 Regs, 2022nov sub). The patients were randomly allocated to the training cohort and the internal validation cohort with a ratio of 7∶3 using bootstrap resampling. The Cox proportional hazards regression analysis was used to determine significant contributors to overall survival (OS) in EAC patients, which would be elected to construct the nomogram prediction model. C-index, calibration curve and receiver operating characteristic (ROC) curve were performed to evaluate its efficacy. Finally, the efficacy to evaluate the OS of EAC patients was compared between the nomogram prediction model and TNM staging system. Results In total, 3945 patients with lower third and abdominal part of EAC were enrolled, including 3475 males and 470 females with a median age of 65 (57-72) years. The 2761 patients were allocated to the training cohort and the remaining 1184 patients to the internal validation cohort. In the training and the internal validation cohorts, the C-index of the nomogram model was 0.705 and 0.713, respectively. Meanwhile, the calibration curve also suggested that the nomogram model had a strong capability of predicting 1-, 3-, and 5-year OS rates of EAC patients. The nomogram also had a higher efficacy than the TNM staging system in predicting 1-, 3-, and 5-year OS rates of EAC patients. Conclusion This nomogram prediction model has a high efficiency for predicting OS in the patients with lower third and abdominal part of EAC, which is higher than that of the current TNM staging system.

Objective To observe the value of right ventricular free wall longitudinal strain(RVFWLS)to pulmonary artery systolic pressure(PASP)ratio(RVFWLS/PASP)for evaluating systolic function of right ventricle(RV)in patients with hypertrophic cardiomyopathy(HCM).Methods Fifty-two HCM patients were retrospectively collected and divided into group A(RVFWLS/PASP≤0.75,n=26)and B(RVFWLS/PASP＞0.75,n=26)according to median RVFWLS/PASP.Meanwhile,26 healthy individuals were included as control group.Parameters of echocardiography and speckle tracking imaging(STI)were obtained and compared among 3 groups.The correlations of RVFWLS/PASP and other parameters were analyzed with linear regression.Results In group A and B,left ventricular ejection fraction(LVEF),RV septal longitudinal strain(RVSepLS),RVFWLS,RV end-diastolic volume index(RVEDVi),RV stroke volume(RVSV),RV fractional area change(RVFAC)and left ventricular global longitudinal strain(LVGLS)were all lower,while the ratio of peak E blood flow velocity at mitral orifice diastole to early mitral intervalal annulus diastole velocity(E/e′)were higher than those in control group(all adjusted P＜0.05).Compared with group B and control group,group A had higher PASP but lower RV ejection fraction(RVEF)(all adjusted P＜0.05).Furthermore,RVFWLS and LVGLS were both lower in group A than in group B(both adjusted P＜0.05).Linear regression analysis showed that RVSepLS and RVEF were independently linearly related to RVFWLS/PASP(both P＜0.05).Conclusion RVFWLS/PASP could be used to monitor changes of RV systoic function in HCM patients at early stage.

Objective:To explore the interaction of IL-2,IL-6 factors and intestinal flora disorder in patients with rheumatoid arthritis complicated with interstitial lung disease(RA-ILD).Methods:A total of 100 RA patients diagnosed at Shijiazhuang People's Hospital from June 2021 to June 2022 were enrolled and divided into the RA-ILD group(with ILD,n=27)and RA-noILD group(with-out ILD,n=73)according to the results of pulmonary CT and clinical manifestations.Fecal samples were collected from all partici-pants,and intestinal flora levels were cultured and analyzed.Bacterial DNA was extracted from fecal samples,and the 16S rDNA gene was amplified by PCR to generate DGGE profiles of gut microbiota.The Shannon-Wiener index,evenness,and richness of intestinal bacteria were calculated.Cluster analysis was performed to compare structural differences in gut microbiota between the groups.Demo-graphic characteristics,clinical symptoms,laboratory parameters,and intestinal flora-related indicators were compared between the groups.Multivariate Logistic regression analysis was conducted to identify independent risk factors for ILD development in RA pa-tients.Spearman correlation analysis was used to evaluate relationships between serum IL-2,IL-6 levels and gut microbiota parame-ters.Cox proportional hazards regression analysis was performed to determine independent factors influencing disease progression in RA-ILD patients.Results:Smoking,age≥53.77 years old,anti-CCP antibody≥334.60 RU/ml,IL-6≥199.47 ng/ml,IL-2<3.10 ng/ml,richness<18.39,Shannon-Wiener index<2.88,Bifidobacterium<7.27 CFU/g,Bacteroides fragilis<7.75 CFU/g,Hmax<3.14 and flora uniformity<0.92 were independent factors affecting the occurrence of ILD in patients with RA.In patients with RA-ILD,serum IL-6 level was negatively correlated with richness,Shannon-Wiener index,Hmax and flora evenness,while serum IL-2 level was positively correlated with richness,Shannon-Wiener index,Hmax and flora evenness.Smoking,age,anti-CCP antibody,IL-6,IL-2,richness,Shannon-Wiener index,Bifidobacterium,Bacteroides fragilis,Hmax and flora uniformity were independent factors affecting the outcome of RA-ILD patients(P<0.05).Conclusion:Smoking,age,anti-CCP antibody,IL-6,IL-2,richness,Shannon-Wiener index,Bifido-bacterium,Bacteroides fragilis,Hmax and flora uniformity have significant diagnostic and prognostic value for RA-ILD.The changes of intestinal microbial community in patients with RA-ILD and the changes in the number and structure of intestinal bacteria may be one of the important factors of RA patients complicated with ILD.

OBJECTIVE: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. METHODS: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. RESULTS: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. CONCLUSION Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

BACKGROUND:Long non-coding RNA(LncRNA)plays an important role in nervous system development and neurological diseases.Previous studies by the research team have demonstrated that human umbilical cord mesenchymal stem cells overexpressing erythropoietin(EPO-MSCs)under ischemic and hypoxic conditions have better neuroprotective functions and significantly activate the expression of LncRNA XIST.Research suggests that XIST is related to the pathogenesis of hypoxic-ischemic encephalopathy,but the role and mechanism of its regulation by EPO-MSCs in protecting ischemic-hypoxic neurons remain unclear.OBJECTIVE:To explore the new mechanism by which LncRNA XIST,in response to EPO-MSC signaling,affects the apoptosis of ischemic-hypoxic SH-SY5Y cells.METHODS:(1)SH-SY5Y cell lines with knockdown of LncRNA XIST(sh-XIST)and negative control(NC-XIST)were constructed through lentiviral transfection.Oxygen-glucose deprivation was used to induce ischemic-hypoxic injury in the cells.Transwell chambers were used to create a non-contact co-culture system with EPO-MSCs,sh-XIST,and NC-XIST ischemic-hypoxic SH-SY5Y cells.Cell proliferation ability was detected using the CCK-8 assay.Cell migration ability was assessed using the scratch assay,and cell apoptosis was measured by flow cytometry.(2)RNA-seq bioinformatics analysis was performed to screen for differentially expressed genes and pathways between sh-XIST and NC-XIST cell lines.Dual-luciferase experiments were used to verify the relationship between miR-124-3p and the target genes XIST and GRIN1.qRT-PCR was conducted to validate the expression levels of downstream miR-124-3p and GRIN1 genes.(3)miR-124-3p inhibitors and mimics were added to verify phenotypic changes in SH-SY5Y cells after ischemic-hypoxic injury and co-culture with EPO-MSCs.RESULTS AND CONCLUSION:(1)Compared with the NC-XIST group,SH-SY5Y cells in the sh-XIST group showed reduced proliferation and migration abilities and increased apoptosis after ischemic-hypoxic injury and co-culture with EPO-MSCs.(2)Dual-luciferase experiments showed that miR-124-3p interacted with the target gene XIST.SH-SY5Y cells transfected with miR-124-3p mimics and co-cultured with EPO-MSCs showed decreased apoptosis after ischemic-hypoxic injury,while SH-SY5Y cells transfected with miR-124-3p inhibitors showed increased apoptosis after co-culture with EPO-MSCs.(3)Transcriptomic sequencing and bioinformatics analysis of sh-XIST revealed significant downregulation of the neuroactive ligand-receptor pathway and the key receptor gene GRIN1 for central nervous system development.(4)Dual-luciferase experiments showed that miR-124-3p interacted with GRIN1.GRIN1 expression was significantly downregulated in the sh-XIST group after ischemic-hypoxic injury compared with the NC-XIST group.These findings indicate that LncRNA XIST promotes GRIN1 expression by upregulating miR-124-3p,thereby reducing cell apoptosis after ischemic-hypoxic injury and co-culture with EPO-MSCs and enhancing proliferation and migration.sh-XIST can block this protective function.

BACKGROUND:Long non-coding RNA(LncRNA)plays an important role in nervous system development and neurological diseases.Previous studies by the research team have demonstrated that human umbilical cord mesenchymal stem cells overexpressing erythropoietin(EPO-MSCs)under ischemic and hypoxic conditions have better neuroprotective functions and significantly activate the expression of LncRNA XIST.Research suggests that XIST is related to the pathogenesis of hypoxic-ischemic encephalopathy,but the role and mechanism of its regulation by EPO-MSCs in protecting ischemic-hypoxic neurons remain unclear.OBJECTIVE:To explore the new mechanism by which LncRNA XIST,in response to EPO-MSC signaling,affects the apoptosis of ischemic-hypoxic SH-SY5Y cells.METHODS:(1)SH-SY5Y cell lines with knockdown of LncRNA XIST(sh-XIST)and negative control(NC-XIST)were constructed through lentiviral transfection.Oxygen-glucose deprivation was used to induce ischemic-hypoxic injury in the cells.Transwell chambers were used to create a non-contact co-culture system with EPO-MSCs,sh-XIST,and NC-XIST ischemic-hypoxic SH-SY5Y cells.Cell proliferation ability was detected using the CCK-8 assay.Cell migration ability was assessed using the scratch assay,and cell apoptosis was measured by flow cytometry.(2)RNA-seq bioinformatics analysis was performed to screen for differentially expressed genes and pathways between sh-XIST and NC-XIST cell lines.Dual-luciferase experiments were used to verify the relationship between miR-124-3p and the target genes XIST and GRIN1.qRT-PCR was conducted to validate the expression levels of downstream miR-124-3p and GRIN1 genes.(3)miR-124-3p inhibitors and mimics were added to verify phenotypic changes in SH-SY5Y cells after ischemic-hypoxic injury and co-culture with EPO-MSCs.RESULTS AND CONCLUSION:(1)Compared with the NC-XIST group,SH-SY5Y cells in the sh-XIST group showed reduced proliferation and migration abilities and increased apoptosis after ischemic-hypoxic injury and co-culture with EPO-MSCs.(2)Dual-luciferase experiments showed that miR-124-3p interacted with the target gene XIST.SH-SY5Y cells transfected with miR-124-3p mimics and co-cultured with EPO-MSCs showed decreased apoptosis after ischemic-hypoxic injury,while SH-SY5Y cells transfected with miR-124-3p inhibitors showed increased apoptosis after co-culture with EPO-MSCs.(3)Transcriptomic sequencing and bioinformatics analysis of sh-XIST revealed significant downregulation of the neuroactive ligand-receptor pathway and the key receptor gene GRIN1 for central nervous system development.(4)Dual-luciferase experiments showed that miR-124-3p interacted with GRIN1.GRIN1 expression was significantly downregulated in the sh-XIST group after ischemic-hypoxic injury compared with the NC-XIST group.These findings indicate that LncRNA XIST promotes GRIN1 expression by upregulating miR-124-3p,thereby reducing cell apoptosis after ischemic-hypoxic injury and co-culture with EPO-MSCs and enhancing proliferation and migration.sh-XIST can block this protective function.

Objective To observe the value of right ventricular free wall longitudinal strain(RVFWLS)to pulmonary artery systolic pressure(PASP)ratio(RVFWLS/PASP)for evaluating systolic function of right ventricle(RV)in patients with hypertrophic cardiomyopathy(HCM).Methods Fifty-two HCM patients were retrospectively collected and divided into group A(RVFWLS/PASP≤0.75,n=26)and B(RVFWLS/PASP＞0.75,n=26)according to median RVFWLS/PASP.Meanwhile,26 healthy individuals were included as control group.Parameters of echocardiography and speckle tracking imaging(STI)were obtained and compared among 3 groups.The correlations of RVFWLS/PASP and other parameters were analyzed with linear regression.Results In group A and B,left ventricular ejection fraction(LVEF),RV septal longitudinal strain(RVSepLS),RVFWLS,RV end-diastolic volume index(RVEDVi),RV stroke volume(RVSV),RV fractional area change(RVFAC)and left ventricular global longitudinal strain(LVGLS)were all lower,while the ratio of peak E blood flow velocity at mitral orifice diastole to early mitral intervalal annulus diastole velocity(E/e′)were higher than those in control group(all adjusted P＜0.05).Compared with group B and control group,group A had higher PASP but lower RV ejection fraction(RVEF)(all adjusted P＜0.05).Furthermore,RVFWLS and LVGLS were both lower in group A than in group B(both adjusted P＜0.05).Linear regression analysis showed that RVSepLS and RVEF were independently linearly related to RVFWLS/PASP(both P＜0.05).Conclusion RVFWLS/PASP could be used to monitor changes of RV systoic function in HCM patients at early stage.

Objective:To explore the interaction of IL-2,IL-6 factors and intestinal flora disorder in patients with rheumatoid arthritis complicated with interstitial lung disease(RA-ILD).Methods:A total of 100 RA patients diagnosed at Shijiazhuang People's Hospital from June 2021 to June 2022 were enrolled and divided into the RA-ILD group(with ILD,n=27)and RA-noILD group(with-out ILD,n=73)according to the results of pulmonary CT and clinical manifestations.Fecal samples were collected from all partici-pants,and intestinal flora levels were cultured and analyzed.Bacterial DNA was extracted from fecal samples,and the 16S rDNA gene was amplified by PCR to generate DGGE profiles of gut microbiota.The Shannon-Wiener index,evenness,and richness of intestinal bacteria were calculated.Cluster analysis was performed to compare structural differences in gut microbiota between the groups.Demo-graphic characteristics,clinical symptoms,laboratory parameters,and intestinal flora-related indicators were compared between the groups.Multivariate Logistic regression analysis was conducted to identify independent risk factors for ILD development in RA pa-tients.Spearman correlation analysis was used to evaluate relationships between serum IL-2,IL-6 levels and gut microbiota parame-ters.Cox proportional hazards regression analysis was performed to determine independent factors influencing disease progression in RA-ILD patients.Results:Smoking,age≥53.77 years old,anti-CCP antibody≥334.60 RU/ml,IL-6≥199.47 ng/ml,IL-2<3.10 ng/ml,richness<18.39,Shannon-Wiener index<2.88,Bifidobacterium<7.27 CFU/g,Bacteroides fragilis<7.75 CFU/g,Hmax<3.14 and flora uniformity<0.92 were independent factors affecting the occurrence of ILD in patients with RA.In patients with RA-ILD,serum IL-6 level was negatively correlated with richness,Shannon-Wiener index,Hmax and flora evenness,while serum IL-2 level was positively correlated with richness,Shannon-Wiener index,Hmax and flora evenness.Smoking,age,anti-CCP antibody,IL-6,IL-2,richness,Shannon-Wiener index,Bifidobacterium,Bacteroides fragilis,Hmax and flora uniformity were independent factors affecting the outcome of RA-ILD patients(P<0.05).Conclusion:Smoking,age,anti-CCP antibody,IL-6,IL-2,richness,Shannon-Wiener index,Bifido-bacterium,Bacteroides fragilis,Hmax and flora uniformity have significant diagnostic and prognostic value for RA-ILD.The changes of intestinal microbial community in patients with RA-ILD and the changes in the number and structure of intestinal bacteria may be one of the important factors of RA patients complicated with ILD.

BACKGROUND:Previous studies have successfully constructed erythropoietin-overexpressed umbilical cord mesenchymal stem cells.It was found that the apoptosis of ischemic and hypoxic human neuroblastoma cell line(SH-SY5Y)was significantly reduced by erythropoietin-overexpressed umbilical cord mesenchymal stem cells. OBJECTIVE:To explore the possible neuroprotective mechanisms of erythropoietin-overexpressed umbilical cord mesenchymal stem cells against ischemic-hypoxic SH-SY5Y and their associated epigenetic mechanisms. METHODS:Oxygen-glucose deprivation was applied to ischemia-hypoxia-induced SH-SY5Y cell injury,and multifactorial assays were applied to detect the expression levels of inflammatory factors in the cells before and after hypoxia and co-culture,respectively,with mesenchymal stem cells,as well as lentiviral-transfected null-loaded plasmids of the negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression levels of supernatant inflammatory factors were detected by multifactor assay after co-culture.Proteomics was used to detect the differentially expressed proteins of negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.Cleavage under targets and tagmentation sequencing was applied to detect genomic H3K4me2 modification,and joint analysis was conducted with RNA-sequencing.Lentiviral vector infection was applied to construct the stable knockdown of REST in SH-SY5Y cells.qRT-PCR and western blot assay were performed to detect the expression level of REST.The apoptosis was detected by flow cytometry after co-culture of oxygen-glucose deprivation treatment with erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression difference of H3K36me3 group proteins was detected by western blot assay,and transcriptome sequencing was performed to analyze the differentially expressed genes. RESULTS AND CONCLUSION:(1)Compared with the control group,monocyte chemotactic protein 1,interleukin-6,interleukin-18,and interleukin-1 beta,interferon α2,and interleukin-23 levels significantly increased in the cerebrospinal fluid supernatant of patients with ischemic-hypoxic encephalopathy(P<0.01).(2)After co-culturing SH-SY5Y cells with erythropoietin-overexpressed umbilical cord mesenchymal stem cells under ischemia and hypoxia,the expression levels of monocyte chemotactic protein 1 and interleukin-6 were significantly reduced.(3)Analysis of protein network interactions revealed significant downregulation of monocyte chemotactic protein 1,interleukin-6 related regulatory proteins CXCL1 and BGN.(4)Transcriptome sequencing analysis found that pro-inflammatory genes were down-regulated,and functional enrichment of histone modifications,and the expression of transcription factors REST and TET3 significantly up-regulated in the erythropoietin-overexpressed umbilical cord mesenchymal stem cell group compared with the negative control mesenchymal stem cell group.(5)Combined analysis of transcriptome sequencing and cleavage under targets and tagmentation revealed changes in epigenetic levels as well as significant activation of the promoter regions of transcription factors REST and TET3.(6)Stable knockdown REST in SH-SY5Y cells was successfully constructed;the transcript levels of REST mRNA and protein expression were both decreased.(7)After the REST knockdown SH-SY5Y cells were co-cultured with erythropoietin-overexpressed umbilical cord mesenchymal stem cells,apoptosis was significantly increased and H3K36me3 expression was significantly decreased.Transcriptome sequencing results showed that the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2 was reduced at mRNA transcription level(P<0.01).(8)It is concluded that erythropoietin-overexpressed umbilical cord mesenchymal stem cells activated the expression of REST and TET3 by altering the kurtosis of H3K4me2 and upregulated the modification level of H3K36me3,which in turn regulated the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2,and facilitated neuronal survival.

Objective:To explore characteristics of intestinal flora in patients with rheumatoid arthritis complicated with inter-stitial lung disease(RA-ILD),to analyze its influencing factors,and to explore improvement of intestinal flora in patients with RA-ILD by different treatment methods.Methods:A total of 100 patients with RA-ILD who visited Shijiazhuang People's Hospital from June 2021 to June 2022 were selected as research objects,and 100 healthy patients in Shijiazhuang People's Hospital during same pe-riod were selected as healthy group.High throughput sequencing was performed on intestinal flora of two groups,OTU flora abundance and α-Diversity,β-diversity,flora distribution and LEfSe difference were analyzed.Another 100 patients with RA-ILD were randomly divided into group A(n=50)and group B(n=50).Group A was treated with leflunomide+nidanib or pifenidone,and group B was treated with leflunomide+nidanib or pifenidone+bifidobacterium triad.Improvement of intestinal flora in two groups after treatment was analyzed.Correlation analysis was used to determine correlation between changes of intestinal flora and clinical characteristics and indicators in RA-ILD patients.Results:OTU flora abundance,α-diversity and β-diversity in RA-ILD patients were lower than that of healthy people.Abundance of Intestinibacter in RA-ILD group was higher than that in healthy group,and abundance of Lactobacillus,Escherichia coli,Klebsiella,Candidatus saccharimonas,Odoribacter and Enterococcus in RA-ILD group were lower than that in healthy group.After treatment,flora structure of group A and group B was improved.Group B was closer to healthy population,and cu-rative effect was significantly better than that of group A.Integinibacter bacteria level was positively correlated with age,RA course,DAS28,CRP,RF and KL-6(P<0.05),and negatively correlated with DLCO(P<0.05).Lactobacillus level was negatively correlated with course of RA,DAS28,CRP,RF,IL-6(P<0.05),and positively correlated with VC(P<0.05);Shigella coli level was negtively correlatied with RA course,ILD course,DAS28,ESR,RF,TNF-α(P<0.05).Klebsiella level was positively correlated with VC and DLCO(P<0.05);Bacterial level of Candida saccharimonas was negatively correlated with DAS28,ESR,CRP,RF,IL-6,TNF-α(P<0.05)and positive correlated with DLCO(P<0.05);Odoribacter bacteria level was negatively correlated with DAS28,CRP,RF,IL-6 and KL-6(P<0.05),and positively correlated with VC(P<0.05);Enterococcus level was negatively correlated with course of RA,ILD,DAS28,ESR,RF and KL-6(P<0.05),while positively correlated with DLCO(P<0.05).Conclusion:Intestinal microflora abundance of RA-ILD patients is significantly lower than that of healthy people,and there aree significant differences in some microflora,which are closely related to clinical characteristics and indicators.Using of bifidobacterium triad in clinical treatment is helpful to improve intestinal flora of RA-ILD patients.