OBJECTIVES: To explore the mechanism of Qingda Granules (QDG) for alleviating brain damage in spontaneously hypertensive rats (SHRs). METHODS: Twelve 5-week-old SHRs were randomized into SHR control group and SHR+QDG group treated with QDG by gavage at the daily dose of 0.9 g/kg for 12 weeks. The control rats, along with 6 age-matched WKY rats, were treated with saline only. Blood pressure changes of the rats were monitored, and pathologies and neuronal apoptosis in the cerebral cortex were examined with HE staining and TUNEL staining. Cerebral cortical expressions of miR-124 and STAT3 mRNA were detected using RT-qPCR, and the protein expressions of NeuN, STAT3, Bcl-2, Bax, and cleaved caspase-3 were detected with immunohistochemistry and Western blotting. In a HT22 cell model of oxygen and glucose deprivation/reoxygenation (OGD/R), the effects of QDG on cell viability and apoptosis, expressions of miR-124 and STAT3 mRNA, and protein expressions of STAT3, Bcl-2, Bax, and cleaved caspase-3 were evaluated using CCK8 assay, Hoechst 33342 staining, RT-qPCR, and Western blotting. RESULTS: Compared with WKY rats, SHRs had significantly elevated systolic blood pressure, diastolic blood pressure and mean arterial pressure with significantly increased neuronal apoptosis in the cerebral cortex, reduced expressions of NeuN, miR-124 and Bcl-2, and enhanced expressions of STAT3, Bax and cleaved caspase-3 (P<0.05). All these changes in the SHRs were significantly ameliorated by treatment with QDG (P<0.05). In the HT22 cell model, QDG treatment obviously reduced OGD/R-induced cell apoptosis, increased the expressions of miR-124 and Bcl-2, and suppressed the elevation of protein expressions of STAT3, Bax and cleaved caspase-3. CONCLUSIONS QDG inhibits cerebral cortical neuronal apoptosis and thereby attenuates brain damage in SHR rats by modulating the miR-124/STAT3 signaling axis.
Animals ; Rats, Inbred SHR ; MicroRNAs/metabolism* ; STAT3 Transcription Factor/metabolism* ; Signal Transduction/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Rats ; Apoptosis/drug effects* ; Rats, Inbred WKY ; Male ; Hypertension

Objective To explore the mechanism of Qingda Granules(QDG)for alleviating brain damage in spontaneously hypertensive rats(SHRs).Methods Twelve 5-week-old SHRs were randomized into SHR control group and SHR+QDG group treated with QDG by gavage at the daily dose of 0.9 g/kg for 12 weeks.The control rats,along with 6 age-matched WKY rats,were treated with saline only.Blood pressure changes of the rats were monitored,and pathologies and neuronal apoptosis in the cerebral cortex were examined with HE staining and TUNEL staining.Cerebral cortical expressions of miR-124 and STAT3 mRNA were detected using RT-qPCR,and the protein expressions of NeuN,STAT3,Bcl-2,Bax,and cleaved caspase-3 were detected with immunohistochemistry and Western blotting.In a HT22 cell model of oxygen and glucose deprivation/reoxygenation(OGD/R),the effects of QDG on cell viability and apoptosis,expressions of miR-124 and STAT3 mRNA,and protein expressions of STAT3,Bcl-2,Bax,and cleaved caspase-3 were evaluated using CCK8 assay,Hoechst 33342 staining,RT-qPCR,and Western blotting.Results Compared with WKY rats,SHRs had significantly elevated systolic blood pressure,diastolic blood pressure and mean arterial pressure with significantly increased neuronal apoptosis in the cerebral cortex,reduced expressions of NeuN,miR-124 and Bcl-2,and enhanced expressions of STAT3,Bax and cleaved caspase-3(P＜0.05).All these changes in the SHRs were significantly ameliorated by treatment with QDG(P＜0.05).In the HT22 cell model,QDG treatment obviously reduced OGD/R-induced cell apoptosis,increased the expressions of miR-124 and Bcl-2,and suppressed the elevation of protein expressions of STAT3,Bax and cleaved caspase-3.Conclusion QDG inhibits cerebral cortical neuronal apoptosis and thereby attenuates brain damage in SHR rats by modulating the miR-124/STAT3 signaling axis.

Objective:To investigate the use of non-vitamin K antagonist oral anticoagulants(NOACs)and their associated comorbidities in patients aged 80 years and older with non-valvular atrial fibrillation(NVAF), as well as to understand the challenges faced by elderly patients receiving NOAC therapy.Methods:We retrospectively enrolled elderly patients(≥80 years old)with NVAF who were treated with NOACs at a hospital in Beijing from January 2018 to August 2023.Patients were categorized into two age groups: 80-89 years and ≥90 years.We collected baseline data, including demographic characteristics, details of atrial fibrillation, comorbidities, laboratory test results, and medication combinations, for descriptive statistical analysis and intergroup comparisons.Results:A total of 695 elderly patients with NVAF receiving NOACs were included in the study, with a median age of 84 years.Among these patients, there were 328 males(47.19%, 328/695)and 422 cases of paroxysmal atrial fibrillation(60.72%, 422/695).The age group of 80-89 years comprised 640 cases(92.09%, 640/695), while the group aged 90 years and above included 55 cases(7.91%, 55/695).The use of NOACs in patients aged 90 and older exhibited an increasing trend over the years.Inter-group comparisons indicated that the ≥90 years group had lower body mass index, longer hospital stays, increased bedridden time, poorer renal function, lower levels of albumin and hemoglobin, and higher D-dimer levels.Inappropriate dosing of DOACs occurred in 49.64%(345/695)of cases, with 90.72%(313/345)receiving doses lower than recommended.Lower-than-recommended doses were more prevalent in the ≥90 years group, while higher-than-recommended doses were more common in the 80-89 years group.Polypharmacy was noted in 61.29%(426/695)of patients.The concurrent use of antiplatelet drugs, rhythm control medications, and ventricular rate control drugs was observed in 12.52%(87/695), 19.57%(136/695), and 54.53%(379/695)of patients, respectively, with no significant differences between groups.Conclusions:Inappropriate dosing and polypharmacy are prevalent issues among elderly NVAF patients.Therefore, it is essential to enhance multidisciplinary collaboration to optimize anticoagulation treatment strategies.

Objective To investigate the mechanism of action and potential biomarkers of Jiangzhi Qingshen capsules in the treatment of dyslipidaemia based on clinical lipid metabolomics.Methods 30 patients with dyslipidaemia from Zhengzhou Hospital of Chinese Medicine were collected as the test group,and their lipid levels before and after taking Jiang Zhi Qing Shen capsule for 12 weeks were compared.Another 30 healthy patients were enrolled in the physical examination department of Zhengzhou Hospital of Chinese Medicine,and metabolomics investigation was carried out on plasma samples of the test group before and after the treatment as well as those of the healthy patients by LC-MS/MS technology.The differences between the groups were compared by multivariate statistical analysis,and the potential biomarkers were identified by HMDB and lipidblast databases,so as to clarify the possible pathways and targets for the treatment of mild dyslipidaemia by Jiangzhi Qingshen capsules.Results Jiangzhi Qingshen capsules could improve patients' blood lipid level,BMI and abdominal circumference significantly(P<0.05).And metabolomics results showed that 29 lipid metabolites in the plasma of the treated patients were dialled back,which involved cholesterol metabolism,fat digestion and absorption,glycerol-phospholipid metabolism and other biological processes.Conclusion The efficacy of Jiangzhi Qingshen capsules in treating dyslipidaemia was confirmed,and its mechanism of action might be related to the regulation of lipid metabolism.Metabolites such as acylcarnitine(Acar),phosphatidylethanolamine(PE)and phosphatidylcholine(PC)were expected to be used as the biomarkers of dyslipidaemia,so as to provide objective scientific basis for the lipid-lowering capsule,and to facilitate the promotion of its application.

Objective To investigate the mechanism of action and potential biomarkers of Jiangzhi Qingshen capsules in the treatment of dyslipidaemia based on clinical lipid metabolomics.Methods 30 patients with dyslipidaemia from Zhengzhou Hospital of Chinese Medicine were collected as the test group,and their lipid levels before and after taking Jiang Zhi Qing Shen capsule for 12 weeks were compared.Another 30 healthy patients were enrolled in the physical examination department of Zhengzhou Hospital of Chinese Medicine,and metabolomics investigation was carried out on plasma samples of the test group before and after the treatment as well as those of the healthy patients by LC-MS/MS technology.The differences between the groups were compared by multivariate statistical analysis,and the potential biomarkers were identified by HMDB and lipidblast databases,so as to clarify the possible pathways and targets for the treatment of mild dyslipidaemia by Jiangzhi Qingshen capsules.Results Jiangzhi Qingshen capsules could improve patients' blood lipid level,BMI and abdominal circumference significantly(P<0.05).And metabolomics results showed that 29 lipid metabolites in the plasma of the treated patients were dialled back,which involved cholesterol metabolism,fat digestion and absorption,glycerol-phospholipid metabolism and other biological processes.Conclusion The efficacy of Jiangzhi Qingshen capsules in treating dyslipidaemia was confirmed,and its mechanism of action might be related to the regulation of lipid metabolism.Metabolites such as acylcarnitine(Acar),phosphatidylethanolamine(PE)and phosphatidylcholine(PC)were expected to be used as the biomarkers of dyslipidaemia,so as to provide objective scientific basis for the lipid-lowering capsule,and to facilitate the promotion of its application.

Objective:To investigate the effects of catalpol(cat)on autophagy and apoptosis of alveolar epithelial cells in-duced by Streptococcus pneumoniae(SP)by regulating adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)/Unc-51-like kinase 1(ULK1)signaling pathway.Methods:Human alveolar epithelial cells A549 were divided into the following groups:control group,SP group(1×108 CFU/ml),cat low,medium,and high doses groups(0.5,1,2 μg/L),AICAR group(2 μg/L cat+1 mmol/L AMPK activator);CCK-8 method and flow cytometry were applied to detect cell proliferation and apoptosis,respectively;monodansylcadaverine(MDC)staining method was applied to detect autophagy in cells;ELISA was applied to detect the levels of IL-6,IL-1β and TNF-α in the cell supernatant;and Western blot was applied to detect the expressions of Bcl-2 associated X protein(Bax),anti apoptotic factor B cell lymphoblastoma 2(Bcl-2),Beclin-1,P62,microtubule-associated protein 1 light chain 3 Ⅰ(LC3 Ⅰ),microtubule-associated protein 1 light chain 3 Ⅱ(LC3 Ⅱ)and AMPK/mTOR/ULK1 pathway proteins in cells.Results:Compared with control group,the activity of A549 cells and the expression levels of Bcl-2,P62,p-mTOR/mTOR pro-teins in SP group were obviously reduced,the apoptosis rate,autophagy fluorescence intensity,levels of IL-6,IL-1β,TNF-α,the protein expression levels of Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin-1,p-AMPK/AMPK,and p-ULK1/ULK1 were obviously increased(P<0.05);compared with the SP group,the activity of A549 cells and the expression levels of Bcl-2,P62,p-mTOR/mTOR proteins in the low,medium,and high dose cat groups were obviously increased,the apoptosis rate,autophagy fluorescence intensity,levels of IL-6,IL-1β,TNF-α,the protein expression levels of Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin-1,p-AMPK/AMPK,and p-UKL1/UKL1 were ob-viously reduced(P<0.05);the AMPK activator AICAR weakened the inhibitory effect of cat on SP induced autophagy and apoptosis in A549 cells.Conclusion:cat can inhibit SP-induced autophagy and apoptosis in A549 cells,and its mechanism of action may be re-lated to regulating the AMPK/mTOR/ULK1 signaling pathway.

Objective:To investigate the effects of catalpol(cat)on autophagy and apoptosis of alveolar epithelial cells in-duced by Streptococcus pneumoniae(SP)by regulating adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)/Unc-51-like kinase 1(ULK1)signaling pathway.Methods:Human alveolar epithelial cells A549 were divided into the following groups:control group,SP group(1×108 CFU/ml),cat low,medium,and high doses groups(0.5,1,2 μg/L),AICAR group(2 μg/L cat+1 mmol/L AMPK activator);CCK-8 method and flow cytometry were applied to detect cell proliferation and apoptosis,respectively;monodansylcadaverine(MDC)staining method was applied to detect autophagy in cells;ELISA was applied to detect the levels of IL-6,IL-1β and TNF-α in the cell supernatant;and Western blot was applied to detect the expressions of Bcl-2 associated X protein(Bax),anti apoptotic factor B cell lymphoblastoma 2(Bcl-2),Beclin-1,P62,microtubule-associated protein 1 light chain 3 Ⅰ(LC3 Ⅰ),microtubule-associated protein 1 light chain 3 Ⅱ(LC3 Ⅱ)and AMPK/mTOR/ULK1 pathway proteins in cells.Results:Compared with control group,the activity of A549 cells and the expression levels of Bcl-2,P62,p-mTOR/mTOR pro-teins in SP group were obviously reduced,the apoptosis rate,autophagy fluorescence intensity,levels of IL-6,IL-1β,TNF-α,the protein expression levels of Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin-1,p-AMPK/AMPK,and p-ULK1/ULK1 were obviously increased(P<0.05);compared with the SP group,the activity of A549 cells and the expression levels of Bcl-2,P62,p-mTOR/mTOR proteins in the low,medium,and high dose cat groups were obviously increased,the apoptosis rate,autophagy fluorescence intensity,levels of IL-6,IL-1β,TNF-α,the protein expression levels of Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin-1,p-AMPK/AMPK,and p-UKL1/UKL1 were ob-viously reduced(P<0.05);the AMPK activator AICAR weakened the inhibitory effect of cat on SP induced autophagy and apoptosis in A549 cells.Conclusion:cat can inhibit SP-induced autophagy and apoptosis in A549 cells,and its mechanism of action may be re-lated to regulating the AMPK/mTOR/ULK1 signaling pathway.

Objective:To investigate the use of non-vitamin K antagonist oral anticoagulants(NOACs)and their associated comorbidities in patients aged 80 years and older with non-valvular atrial fibrillation(NVAF), as well as to understand the challenges faced by elderly patients receiving NOAC therapy.Methods:We retrospectively enrolled elderly patients(≥80 years old)with NVAF who were treated with NOACs at a hospital in Beijing from January 2018 to August 2023.Patients were categorized into two age groups: 80-89 years and ≥90 years.We collected baseline data, including demographic characteristics, details of atrial fibrillation, comorbidities, laboratory test results, and medication combinations, for descriptive statistical analysis and intergroup comparisons.Results:A total of 695 elderly patients with NVAF receiving NOACs were included in the study, with a median age of 84 years.Among these patients, there were 328 males(47.19%, 328/695)and 422 cases of paroxysmal atrial fibrillation(60.72%, 422/695).The age group of 80-89 years comprised 640 cases(92.09%, 640/695), while the group aged 90 years and above included 55 cases(7.91%, 55/695).The use of NOACs in patients aged 90 and older exhibited an increasing trend over the years.Inter-group comparisons indicated that the ≥90 years group had lower body mass index, longer hospital stays, increased bedridden time, poorer renal function, lower levels of albumin and hemoglobin, and higher D-dimer levels.Inappropriate dosing of DOACs occurred in 49.64%(345/695)of cases, with 90.72%(313/345)receiving doses lower than recommended.Lower-than-recommended doses were more prevalent in the ≥90 years group, while higher-than-recommended doses were more common in the 80-89 years group.Polypharmacy was noted in 61.29%(426/695)of patients.The concurrent use of antiplatelet drugs, rhythm control medications, and ventricular rate control drugs was observed in 12.52%(87/695), 19.57%(136/695), and 54.53%(379/695)of patients, respectively, with no significant differences between groups.Conclusions:Inappropriate dosing and polypharmacy are prevalent issues among elderly NVAF patients.Therefore, it is essential to enhance multidisciplinary collaboration to optimize anticoagulation treatment strategies.

Objective:To investigate the application of (4R)-4-(3-[ 18F]fluoranyl-5-fluorophenyl)-1-((3-methylpyridin-4-yl)methyl)pyrrolidin-2-one( 18F-SynVesT-1), a synaptic vesicle glycoprotein 2A (SV2A) radioactive tracer, in patients with Alzheimer′s disease (AD). Methods:A total of 20 AD patients (2 males, 18 females, age (66.4±8.1) years) with positive β-amyloid (Aβ) deposition and 20 normal controls (NC; 9 males, 11 females, age (62.6±8.6) years ) without Aβ deposition were retrospectively recruited from Huashan Hospital, Fudan University between December 2021 and December 2022. All of them underwent 18F-SynVesT-1 PET/MR and 18F-Florbetapir (AV45) PET/CT scans. Preprocessing of brain 18F-SynVesT-1 PET images was carried out using statistical parametric mapping (SPM). The differences of the uptke of 18F-SynVesT-1 (synaptic density) between two groups based on ROI were compared by using either the independent-sample t test or Mann-Whitney U test. Spearman rank correlation analysis was performed to assess the relationship between synaptic density and cognitive performance. For voxelwise analysis, a general linear model was constructed to analyze differences in synaptic density between the two groups using the independent-sample t test. Furthermore, a multiple linear regression model was developed to explore the relationship between synaptic density and cognitive performance. Results:Compared to the NC group, the AD group exhibited significant widespread reduction in synaptic density across the cortical regions ( P<0.05, false discovery rate (FDR)-corrected), particularly in the medial temporal lobe (0.84±0.09 vs 1.04±0.09; t=-6.95, P<0.001), lateral temporal lobe (1.15±0.13 vs 1.31±0.08; t=-4.56, P<0.001), and lateral parietal lobe (1.24(1.04, 1.26) vs 1.32(1.23, 1.39); z=-3.25, P=0.001). Moreover, synaptic density in extensive cortical regions showed a positive correlation with mini-mental state examination (MMSE) and Montreal cognitive assessment-basic (MoCA-B) scores ( P<0.05, FDR-corrected). Notably, significant associations were observed between MMSE and MoCA-B scores and synaptic density in the lateral temporal lobe ( rs values: 0.71, 0.74, both P<0.001) and medial temporal lobe ( rs values: 0.71, 0.74, both P<0.001). Conclusions:18F-SynVesT-1 PET imaging is a valuable tool for evaluating synaptic density, specifically in the context of AD. The observed widespread reduction in synaptic density across cortical regions of patients with AD are closely related to cognitive decline.

Objective To explore the correlation between the rs290487 and rs7903146 single-nucleotide polymorphism(SNP)of TCF7L2 gene and the occurrence of diabetic kidney disease(DKD)in type 2 diabetes mellitus(T2DM)population in Hebei region.Methods Epidemiological and clinical data as well as SNP data at TCF7L2 rs290487 and rs7903146 sites were collected from 317 non DKD patients and 302 DKD patients.The correlation between SNP at rs290487 and rs7903146 sites and DKD was analyzed through a case-control design.Results The genotype frequency distribution at the rs290487 and rs7903146 loci of the two TCF7L2 genes conforms to the Hardy-Weinberg genetic balance(P>0.05),and the allele frequency distribution conforms to the Hardy-Weinberg genetic balance(P>0.05).Single factor logistic regression analysis of genetic models for different genotypes at rs290487 locus showed that the risk of DKD in TC+CC genotype carriers was 2.772 times that of TT genotype carriers in the dominant model.Multivariate logistic regression analysis of genetic models for different genotypes at rs290487 locus showed that the risk of DKD in TC+CC genotype carriers was 2.837 times that of TT genotype carriers.Conclusions TC and CC genotypes at rs290487 sites of TCF7L2 gene are risk factors for DKD,and gene mutation at rs290487 sites may be associated with DKD in T2DM population in Hebei.