Objective To explore the protective effect and related mechanism of Compound Baimai Powder(CBMP,a compound description of Mongolian medicine)on astrocytes after oxygen glu-cose deprivation and reoxygenation(OGD/R)injury.Methods Astrocyte model of OGD/R injury was subjected to simulate in vitro cerebral ischemia/reperfusion injury.Cultured astrocytes were randomly divided into normal group,OGD/R group,OGD/R+nimodipine group(10 μmol/L),OGD/R+low-and high-dose CBMP groups(25,50 μmol/L).Cell viability and apoptosis were de-tected with CCK-8 assay and flow cytometry,respectively.Western blotting was used to measure the expression levels of the proteins related to the nuclear factor erythroid-2-related factor-2(NRF2)/antioxidant response element and Janus kinase(J AK)/signal transducer and activator of transcription(STAT)signaling pathways.ELISA was employed to examine the levels of inflam-matory factors IL-1β,IL-6,and TNF-α,as well as oxidative stress molecules ROS,GSH,MDA and SOD.Results Compared to the normal group,the OGD/R group showed significant decreases in cell viability,NRF2 protein level,and SOD and GSH activities(P＜0.05,P＜0.01),and obvious increases in p-JAK and p-STAT proteins levels,contents of IL-1β,IL-6 and TNF-α,and ROS and MDA levels(P＜0.05,P＜0.01).High-dose CBMP treatment resulted in notably elevated cell via-bility and NRF2 protein level,while reduced levels of p-JAK[(1.20±0.20)vs(2.50±0.26)]and p-STAT[(1.15±0.25)vs(2.10±0.21)]proteins,IL-6[(30.33±5.20)vs(180.35±18.50)]and TNF-α[(50.12±8.24)vs(160.45±15.20)]when compared to the OGD/R group(P＜0.05,P＜0.01).Conclusion CBMP exerts protective effect on astrocytes against OGD/R injury.

Objective To explore the effect and mechanism of Zuogui Jiangtang Jieyu Formula(ZGJTJYF)in treating diabetes-related depression by regulating glutamate receptor 2(GluR2).Methods The primary isolated and cultured hippocampal neurons of SD rats were used.The experiment consisted of normal,model,blank serum(10％blank serum),positive drug(10％[metformin+fluoxetine]drug-containing serum),20％ZGJTJYF group,10％ZGJTJYF group,10％ZGJTJYF+GluR2 knockdown group,and 10％ZGJTJYF+GluR2 overexpression group(with corresponding volume fractions of ZGJTJYF drug-containing serum added).The ZGJTJYF+GluR2 knockdown and overexpression groups,were transfected with lentivirus to obtain hippocampal neurons with either GluR2 overexpression or knockdown.The glucose(150 mmol/L)and corticosterone(200 μmol/L)were used for 18 h to establish an in vitro cell model of hippocampal neurons in diabetes-related depression.After 24 h of successful modeling,the corresponding serum was added to each group for intervention.After 24 h of intervention,the morphological structure of hippocampal neurons was observed using an optical microscope.Biochemical methods were used to determine the glucose and insulin content in cell supernatant.An enzyme-linked immunosorbent assay was used to detect 5-hydroxytryptamine(5-HT)and dopamine(DA)levels in the cell supernatant,and the microtubule-associated protein 1A/1B light chain 3 autophagy double-labeled adenovirus(mRFP-GFP-LC3)autophagy fluorescence double labeling method was used to detect the average fluorescence intensity of LC3 protein in hippocampal neurons.Nissl staining was used to observe synaptic damage in hippocampal neurons,and an immunofluorescence method was used to detect the protein expression of Parkin,phosphatase and tensin homolog-induced putative kinase 1(PINK1),regulating synaptic membrane exocytosis 3(RIMS3),synapsin 1(SYN1),postsynaptic density-95(PSD-95),synapse-associated protein 102(SAP 102),and GluR2 in hippocampal neurons.Realtime fluorescence PCR was used to detect GluR2 mRNA expression in hippocampal neurons,while Western blotting was employed to assess the expression of mitophagy proteins Parkin and PINK1 in these neurons.Results Compared to the normal group,the model group and blank serum group showed structural damage to hippocampal neurons,increased glucose content in cell supernatant,decreased insulin,5-HT,and DA content,increased average fluorescence intensity of LC3,Parkin,and PINK1,decreased average fluorescence intensity of RIMS3,SYN1,PSD-95,SAP 102,and GluR2,decreased GluR2 mRNA expression,increased protein expression of Parkin and PINK1(P＜0.05),and decreased Nissl bodies.Compared to the model group and blank serum group,the above indicators in each administration group were improved to varying degrees(P＜0.05).Compared to the positive drug group,the average fluorescence intensity of LC3,Parkin,and PINK1 decreased,Parkin and PINK1 protein expression decreased,and the average fluorescence intensity of GluR2,SYN1,and PSD-95 increased in 10％ZGJTJYF,20％ZGJTJYF group,and 10％ZGJTJYF+GluR2 overexpression group(P＜0.05).Compared to 10％and 20％ZGJTJYF groups,10％ZGJTJYF+GluR2 knockdown group showed a decrease in 5-HT content,an increase in average fluorescence intensity of LC3 and Parkin,a decrease in average fluorescence intensity of SYN1,PSD-95,and GluR2,a decreased in GluR2 mRNA expression,and an increase of Parkin and PINK1 protein expression(P＜0.05).In contrast,the above indicators were improved to varying degrees in 10％ZGJTJYF+GluR2 overexpression group(P＜0.05).Compared to 10％ZGJTJYF+GluR2 knockdown group,the above abnormal indicators in 10％ZGJTJYF+GluR2 overexpression group were reversed to varying degrees(P＜0.05).Conclusion ZGJTJYF has a protective effect on synaptic damage of hippocampal neurons in diabetes-related depression,and its mechanism may be related to the upregulation of GluR2 and the inhibition of mitophagy over activation.

Objective To explore the application value of CMR myocardial strain technique in eld-erly males with HCM.Methods A retrospective analysis was conducted on 50 elderly male pa-tients who underwent CMR examination at the Second Medical Center of Chinese PLA General Hospital from April 2020 to December 2024.According to being diagnosed with HCM or not,they were divided into a HCM group(24 cases)and a control group(26 cases).Post-processing soft-ware CVI42 was used to obtain two sets of basic functional and strain parameters of the left ven-tricle(LV)and left atrium(LA).The parameters of LV included LV mass,LV mass index(LV massi),global longitudinal strain(GLS),and so on,while the parameters of LA included minimum LA volume(LAV),minimum LA volume index(LA VI),LA passive strain(εe),LA peak early negative strain rate(SRe)and LA peak late negative strain rate(SRa),etc.The two sets of LV and LA parameters of basic functional and strain were compared between the two groups.Results The HCM group had significant higher LV mass and LV massi and minimum LAV and LAVI,but lower GLS,εe,SRe and SRa in 2D and 3D speckle-tracking echocardiography than the control group(P＜0.05,P＜0.01).The AUC value of 2D GLS,SRa,and logistic regres-sion model in predicting HCM was 0.887(95％CI:0.766-0.959),0.740(95％CI:0.597-0.854),and 0.929(95％CI:0.820-0.983),respectively,with a sensitivity of 76.92％,57.69％,and 84.62％,and a specificity of 70.83％,83.33％,and 91.67％,respectively.The logistic regression model demonstrated higher AUC value,sensitivity,and specificity than 2D GLS and SRa.Conclusion CMR myocardial strain technology is of significant diagnostic value for elderly male patients with HCM.

Objective To explore the effect and mechanism of Zuogui Jiangtang Jieyu Formula(ZGJTJYF)in treating diabetes-related depression by regulating glutamate receptor 2(GluR2).Methods The primary isolated and cultured hippocampal neurons of SD rats were used.The experiment consisted of normal,model,blank serum(10％blank serum),positive drug(10％[metformin+fluoxetine]drug-containing serum),20％ZGJTJYF group,10％ZGJTJYF group,10％ZGJTJYF+GluR2 knockdown group,and 10％ZGJTJYF+GluR2 overexpression group(with corresponding volume fractions of ZGJTJYF drug-containing serum added).The ZGJTJYF+GluR2 knockdown and overexpression groups,were transfected with lentivirus to obtain hippocampal neurons with either GluR2 overexpression or knockdown.The glucose(150 mmol/L)and corticosterone(200 μmol/L)were used for 18 h to establish an in vitro cell model of hippocampal neurons in diabetes-related depression.After 24 h of successful modeling,the corresponding serum was added to each group for intervention.After 24 h of intervention,the morphological structure of hippocampal neurons was observed using an optical microscope.Biochemical methods were used to determine the glucose and insulin content in cell supernatant.An enzyme-linked immunosorbent assay was used to detect 5-hydroxytryptamine(5-HT)and dopamine(DA)levels in the cell supernatant,and the microtubule-associated protein 1A/1B light chain 3 autophagy double-labeled adenovirus(mRFP-GFP-LC3)autophagy fluorescence double labeling method was used to detect the average fluorescence intensity of LC3 protein in hippocampal neurons.Nissl staining was used to observe synaptic damage in hippocampal neurons,and an immunofluorescence method was used to detect the protein expression of Parkin,phosphatase and tensin homolog-induced putative kinase 1(PINK1),regulating synaptic membrane exocytosis 3(RIMS3),synapsin 1(SYN1),postsynaptic density-95(PSD-95),synapse-associated protein 102(SAP 102),and GluR2 in hippocampal neurons.Realtime fluorescence PCR was used to detect GluR2 mRNA expression in hippocampal neurons,while Western blotting was employed to assess the expression of mitophagy proteins Parkin and PINK1 in these neurons.Results Compared to the normal group,the model group and blank serum group showed structural damage to hippocampal neurons,increased glucose content in cell supernatant,decreased insulin,5-HT,and DA content,increased average fluorescence intensity of LC3,Parkin,and PINK1,decreased average fluorescence intensity of RIMS3,SYN1,PSD-95,SAP 102,and GluR2,decreased GluR2 mRNA expression,increased protein expression of Parkin and PINK1(P＜0.05),and decreased Nissl bodies.Compared to the model group and blank serum group,the above indicators in each administration group were improved to varying degrees(P＜0.05).Compared to the positive drug group,the average fluorescence intensity of LC3,Parkin,and PINK1 decreased,Parkin and PINK1 protein expression decreased,and the average fluorescence intensity of GluR2,SYN1,and PSD-95 increased in 10％ZGJTJYF,20％ZGJTJYF group,and 10％ZGJTJYF+GluR2 overexpression group(P＜0.05).Compared to 10％and 20％ZGJTJYF groups,10％ZGJTJYF+GluR2 knockdown group showed a decrease in 5-HT content,an increase in average fluorescence intensity of LC3 and Parkin,a decrease in average fluorescence intensity of SYN1,PSD-95,and GluR2,a decreased in GluR2 mRNA expression,and an increase of Parkin and PINK1 protein expression(P＜0.05).In contrast,the above indicators were improved to varying degrees in 10％ZGJTJYF+GluR2 overexpression group(P＜0.05).Compared to 10％ZGJTJYF+GluR2 knockdown group,the above abnormal indicators in 10％ZGJTJYF+GluR2 overexpression group were reversed to varying degrees(P＜0.05).Conclusion ZGJTJYF has a protective effect on synaptic damage of hippocampal neurons in diabetes-related depression,and its mechanism may be related to the upregulation of GluR2 and the inhibition of mitophagy over activation.

Objective To explore the application value of CMR myocardial strain technique in eld-erly males with HCM.Methods A retrospective analysis was conducted on 50 elderly male pa-tients who underwent CMR examination at the Second Medical Center of Chinese PLA General Hospital from April 2020 to December 2024.According to being diagnosed with HCM or not,they were divided into a HCM group(24 cases)and a control group(26 cases).Post-processing soft-ware CVI42 was used to obtain two sets of basic functional and strain parameters of the left ven-tricle(LV)and left atrium(LA).The parameters of LV included LV mass,LV mass index(LV massi),global longitudinal strain(GLS),and so on,while the parameters of LA included minimum LA volume(LAV),minimum LA volume index(LA VI),LA passive strain(εe),LA peak early negative strain rate(SRe)and LA peak late negative strain rate(SRa),etc.The two sets of LV and LA parameters of basic functional and strain were compared between the two groups.Results The HCM group had significant higher LV mass and LV massi and minimum LAV and LAVI,but lower GLS,εe,SRe and SRa in 2D and 3D speckle-tracking echocardiography than the control group(P＜0.05,P＜0.01).The AUC value of 2D GLS,SRa,and logistic regres-sion model in predicting HCM was 0.887(95％CI:0.766-0.959),0.740(95％CI:0.597-0.854),and 0.929(95％CI:0.820-0.983),respectively,with a sensitivity of 76.92％,57.69％,and 84.62％,and a specificity of 70.83％,83.33％,and 91.67％,respectively.The logistic regression model demonstrated higher AUC value,sensitivity,and specificity than 2D GLS and SRa.Conclusion CMR myocardial strain technology is of significant diagnostic value for elderly male patients with HCM.

Objective To explore the protective effect and related mechanism of Compound Baimai Powder(CBMP,a compound description of Mongolian medicine)on astrocytes after oxygen glu-cose deprivation and reoxygenation(OGD/R)injury.Methods Astrocyte model of OGD/R injury was subjected to simulate in vitro cerebral ischemia/reperfusion injury.Cultured astrocytes were randomly divided into normal group,OGD/R group,OGD/R+nimodipine group(10 μmol/L),OGD/R+low-and high-dose CBMP groups(25,50 μmol/L).Cell viability and apoptosis were de-tected with CCK-8 assay and flow cytometry,respectively.Western blotting was used to measure the expression levels of the proteins related to the nuclear factor erythroid-2-related factor-2(NRF2)/antioxidant response element and Janus kinase(J AK)/signal transducer and activator of transcription(STAT)signaling pathways.ELISA was employed to examine the levels of inflam-matory factors IL-1β,IL-6,and TNF-α,as well as oxidative stress molecules ROS,GSH,MDA and SOD.Results Compared to the normal group,the OGD/R group showed significant decreases in cell viability,NRF2 protein level,and SOD and GSH activities(P＜0.05,P＜0.01),and obvious increases in p-JAK and p-STAT proteins levels,contents of IL-1β,IL-6 and TNF-α,and ROS and MDA levels(P＜0.05,P＜0.01).High-dose CBMP treatment resulted in notably elevated cell via-bility and NRF2 protein level,while reduced levels of p-JAK[(1.20±0.20)vs(2.50±0.26)]and p-STAT[(1.15±0.25)vs(2.10±0.21)]proteins,IL-6[(30.33±5.20)vs(180.35±18.50)]and TNF-α[(50.12±8.24)vs(160.45±15.20)]when compared to the OGD/R group(P＜0.05,P＜0.01).Conclusion CBMP exerts protective effect on astrocytes against OGD/R injury.

Objective:To investigate protective effect of wogonin on myocardial ischemia/reperfusion(MI/R)injury in rats,and to analyze its potential mechanism.Methods:SD rats were randomly separated into sham operation group,MI/R group,trimetazi-dine group(10 mg/kg),wogonin low,medium and high doses groups(5,10,20 mg/kg),with 12 rars per group.MI/R model was established by ligation of left anterior descending coronary artery for 30 min and reperfusion for 24 h.Echocardiography was applied to detect cardiac function in rats,ELISA was applied to detect activities of serum lactate dehydrogenase(LDH),creatine kinase isoen-zyme(CK-MB)and levels of TNF-α,IL-6 and IL-1β in rats;TTC staining was applied to detect myocardial infarct size;HE staining was applied to detect myocardial histopathological changes;TUNEL staining was applied to detect apoptosis level of cardiomyocytes;immunohistochemistry(IHC)method was applied to detect expression of nuclear factor-κB(NF-κB)p65 in myocardial tissue;Western blot was applied to detect myocardial apoptosis and expressions of NF-κB/mitogen-activated protein kinase(MAPK)pathway-related proteins.Results:Compared with sham operation group,LVESD,LVEDD,serum CK-MB and LDH activities,levels of IL-6,IL-1β and TNF-α,myocardial infarction area percentage,myocardial cell apoptosis rate,protein level of Bax,expression of nuclear NF-κB p65,and Cleaved-Caspase-3/Caspase-3,p-P38 MAPK/P38 MAPK in MI/R group were obviously increased,EF,FS and pro-tein levels of Bcl-2 were obviously decreased(P<0.05),myocardial fibers were disordered,and a large number of inflammatory cells were infiltrated;compared with MI/R group,LVESD,LVEDD,serum CK-MB and LDH activities,levels of IL-6,IL-1β and TNF-α,myocardial infarction area percentage,myocardial cell apoptosis rate,protein level of Bax,expression of nuclear NF-κB p65,and Cleaved-Caspase-3/Caspase-3 and p-P38 MAPK/P38 MAPK in trimetazidine group and wogonin high,medium and low doses groups were obviously decreased,EF,FS and protein levels of Bcl-2 were obviously increased(P<0.05),myocardial inflammatory cell infil-tration and muscle fiber fragmentation were reduced in a dose-dependent manner.Conclusion:Wogonin may improve MI/R injury in rats by inhibiting NF-κB/MAPK signaling pathway,inhibiting inflammatory response and cardiomyocyte apoptosis.

Objective:To investigate the influences of Pinocembrin on inflammatory injury in rats with acute myocardial in-farction(AMI)by regulating TLR4/NF-κB/NLRP3 signaling pathway.Methods:The AMI model was established by coronary liga-tion,and the rats were grouped into Sham group,AMI group,Pinocembrin group(5 mg/kg tail vein injection),TLR4 inhibitor group(TAK-242 group,2.0 mg/kg tail vein injection),the levels of cardiac function indexes(LVEF,LVEDD,LVESD,FS)and serum LDH,cTnⅠ,IL-6,IL-β and TNF-α were detected in rats,TTC staining,HE staining and Masson staining were applied to observe myocardial infarction and myocardial histopathological changes in rats,cardiomyocyte apoptosis was detected by TUNEL method,im-munohistochemistry and Western blot were applied to detect TLR4/NF-κB/NLRP3 pathway-related proteins in rat myocardial tissue.Results:Compared with Sham group,the myocardial infarction area increased,the number of myocardial cells decreased,some myo-cardial fibers were broken,inflammatory cells infiltrated,collagen fibers increased,and the apoptosis rate was obviously increased in AMI group(P<0.05),LVEDD,LVESD,serum LDH,cTnⅠ,IL-6,IL-β,TNF-α levels,myocardial tissue TLR4,MyD88,p-NF-κB p65,NLRP3,Caspase-1 expression levels were obviously increased(P<0.05),while LVEF and FS were obviously decreased(P<0.05);compared with AMI group,the myocardial infarction area of the Pinocembrin group and the TAK-242 group were reduced,the cell damage and inflammatory infiltration were reduced,the necrotic cells were obviously reduced,and the apoptosis rate was obvious-ly reduced(P<0.05),LVEDD,LVESD,serum LDH,cTnⅠ,IL-6,IL-β,TNF-α levels,myocardial tissue TLR4,MyD88,p-NF-κB p65,NLRP3,Caspase-1 expression levels were decreased(P<0.05),LVEF and FS were obviously increased(P<0.05);there was no obvious difference in each index between Pinocembrin group and TAK-242 group(P>0.05).Conclusion:Pinocembrin may at-tenuate myocardial inflammatory injury caused by AMI by inhibiting TLR4/NF-κB/NLRP3 signaling pathway.

Serine/arginine-rich splicing factor 7 (SRSF7), a known splicing factor, has been revealed to play oncogenic roles in multiple cancers. However, the mechanisms underlying its oncogenic roles have not been well addressed. Here, based on N⁶-methyladenosine (m⁶A) co-methylation network analysis across diverse cell lines, we find that the gene expression of SRSF7 is positively correlated with glioblastoma (GBM) cell-specific m⁶A methylation. We then indicate that SRSF7 is a novel m⁶A regulator, which specifically facilitates the m⁶A methylation near its binding sites on the mRNAs involved in cell proliferation and migration, through recruiting the methyltransferase complex. Moreover, SRSF7 promotes the proliferation and migration of GBM cells largely dependent on the presence of the m⁶A methyltransferase. The two m⁶A sites on the mRNA for PDZ-binding kinase (PBK) are regulated by SRSF7 and partially mediate the effects of SRSF7 in GBM cells through recognition by insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). Together, our discovery reveals a novel role of SRSF7 in regulating m⁶A and validates the presence and functional importance of temporal- and spatial-specific regulation of m⁶A mediated by RNA-binding proteins (RBPs).
Humans ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Glioblastoma/genetics* ; Methyltransferases/metabolism* ; RNA Splicing Factors/metabolism* ; RNA, Messenger/genetics* ; RNA-Binding Proteins/metabolism* ; Serine-Arginine Splicing Factors/metabolism* ; RNA Methylation/genetics*

BACKGROUND: Anorexia nervosa (AN) is a psychological disorder, which is characterized by the misunderstanding of body image, food restriction, and low body weight. An increasing number of studies have reported that the pathophysiological mechanism of AN might be associated with the dysbiosis of gut microbiota. The purpose of our study was to explore the features of gut microbiota in patients with AN, hoping to provide valuable information on its pathogenesis and treatment. METHODS: In this cross-sectional study, from August 2020 to June 2021, patients with AN who were admitted into Peking University Third Hospital and Peking University Sixth Hospital ( n   =  30) were recruited as the AN group, and healthy controls (HC) were recruited from a middle school and a university in Beijing ( n   =  30). Demographic data, Hamilton Depression Scale (HAMD) scores of the two groups, and length of stay of the AN group were recorded. Microbial diversity analysis of gut microbiota in stool samples from the two groups was analyzed by 16S ribosomal RNA (rRNA) gene sequencing. RESULTS: The weight (AN vs. HC, [39.31 ± 7.90] kg vs. [56.47 ± 8.88] kg, P  < 0.001) and body mass index (BMI, AN vs. HC, [14.92 ± 2.54] kg/m 2vs. [20.89 ± 2.14] kg/m 2 , P  < 0.001) of patients with AN were statistically significantly lower than those of HC, and HAMD scores in AN group were statistically significantly higher than those of HC. For alpha diversity, there were no statistically significant differences between the two groups; for beta diversity, the two groups differed obviously regarding community composition. Compared to HC, the proportion of Lachnospiraceae in patients with AN was statistically significantly higher (AN vs. HC, 40.50% vs. 31.21%, Z  = -1.981, P  = 0.048), while that of Ruminococcaceae was lower (AN vs. HC, 12.17% vs. 19.15%, Z  = -2.728, P  = 0.007); the proportion of Faecalibacterium (AN vs. HC, 3.97% vs. 9.40%, Z  = -3.638, P  < 0.001) and Subdoligranulum (AN vs. HC, 4.60% vs. 7.02%, Z  = -2.369, P  = 0.018) were statistically significantly lower, while that of Eubacterium_hallii_group was significantly higher (AN vs. HC, 7.63% vs. 3.43%, Z  = -2.115, P  = 0.035). Linear discriminant effect (LEfSe) analysis (LDA score >3.5) showed that o_Lachnospirales, f_Lachnospiraceae, and g_Eubacterium_hallii_group (o, f and g represents order, family and genus respectively) were enriched in patients with AN. Microbial function of nutrient transport and metabolism in AN group were more abundant ( P  > 0.05). In AN group, weight and BMI were significantly negatively correlated with the abundance of Bacteroidota and Bacteroides , while positively correlated with Subdoligranulum . BMI was significantly positively correlated with Firmicutes; HAMD scores were significantly negatively correlated with Faecalibacterium. CONCLUSIONS The composition of gut microbiota in patients with AN was different from that of healthy people. Clinical indicators have correlations with the abundance of gut microbiota in patients with AN.
Humans ; Gastrointestinal Microbiome/physiology* ; Anorexia Nervosa ; Cross-Sectional Studies ; Dysbiosis/microbiology* ; Body Mass Index ; RNA, Ribosomal, 16S/genetics* ; Feces/microbiology*