Abstract:Objective:To screen the susceptible genes of patients with early-onset hypertension combined with abnormal potassium metabolism based on whole-exome sequencing and bioinformatics analysis,and to explore the potential molecular mechanism.Methods:A total of 57 patients with early-onset hypertension were included.According to the blood potassium at the time of admission,the patients were divided into the normal potassium group(blood potassium 3.5-5.5 mmol/L,21 cases)and the low potassium group(blood potassium<3.5 mmol/L,36 cases).Peripheral blood samples were collected for whole exome sequencing and variant annotation,and the genes with interactive differences between the two groups were screened.Gene ontology(GO)functional enrichment and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed using the DAVID platform.The protein-protein interaction(PPI)network was constructed using the STRING database and imported into Cytoscape software for topological analysis to identify key node genes and functional modules.The protein expressions of plasma type Ⅳ collagen α3 chain(COL4A3)and type Ⅳ collagen α5 chain(COL4A5)in the two groups were detected by enzyme-linked immunosorbent assay(ELISA).Results:There were statistically differences in term of age,early diastolic rapid filling velocity of the left ventricle(E peak),the ratio of early diastolic rapid filling velocity of the left ventricle to late diastolic filling velocity of the left ventricle(A peak)(E/A),blood potassium and blood calcium levels,and the proportion of stroke/cerebral infarction between the two groups(P<0.05).A total of 63 interactive differential genes were identified through genetic screening.The PPI network topological analysis revealed that genes such as fibronectin 1(FN1),potassium voltage-gated channel subfamily Q member 1(KCNQ1),myosin(TTN),neurofibromatosis type 1(NF1),lysine methyltransferase 2D(KMT2D),insulin-like growth factor 2(IGF2),Barttin protein(BSND),calcium voltage-gated channel subfamily α1 H subunit(CACNA1H),sodium channel epithelial type 1 regulatory subunit γ(SCNN1G),COL4A3 and COL4A5 showed higher betweenness centrality(BC).The molecular complex detection(MCODE)clustering analysis revealed that FN1 and COL4A3,type Ⅳ collagen α4 chain(COL4A4),COL4A5,and type Ⅴcollagen α1 chain(COL5A1)formed a functional module of tight junction,which was significantly enriched in the integrin signaling pathway.The plasma levels of COL4A3 and COL4A5 in the low blood potassium group decreased more than those in the normal blood potassium group(P<0.05).Conclusion:Through exome sequencing and bioinformatics integration analysis,potential susceptibility genes of early-onset hypertension combined with abnormal potassium metabolism were screened out,including FN1,KMT2D,NF1,TTN,COL4A3,and COL4A5.COL4A3 and COL4A5 might be key molecules involved in the occurrence and development of the disease through the integrin signaling pathway.It might be theoretical basis of early-onset hypertension.

Abstract:Objective:To observe the effect of acupuncture at Neiguan(PC6)-Jueyinshu(BL14)combined with oral western medicine on the circadian rhythm of blood pressure in patients with non-dipper hypertension.Methods:Sixty patients with non-dipper hypertension were randomly divided into the control group and the treatment group,with 30 cases in each group.The control group was received conventional treatment(health education combined with oral western medicine),while the treatment group was received additional acupuncture treatment on the basis of the control group.The selected acupoints were Neiguan(PC6,bilateral),Jueyinshu(BL14,bilateral),Taichong(LR3,bilateral),and Baihui(GV20).Two groups were treated for 8 weeks.Results:The total effective rate of circadian rhythm improvement of blood pressure in the treatment group was 80.00％,which was significantly higher than 36.67％in the control group,and the difference was statistically significant(P<0.001).After treatment,the nocturnal blood pressure reduction rate in the treatment group was significantly higher than that before treatment(P<0.05),and also higher than that in the control group after treatment(P<0.05).After treatment,the 24 h mean systolic blood pressure(24 h-MSBP),daytime mean systolic blood pressure(d-MSBP),and nighttime mean systolic blood pressure(n-MSBP)in both groups were significantly lower than those before treatment(P<0.05),the n-MSBP in the treatment group after treatment was lower than that in the control group(P<0.05).After treatment,the 24 h mean diastolic blood pressure(24 h-MDBP),daytime mean diastolic blood pressure(d-MDBP),and nighttime mean diastolic blood pressure(n-MDBP)in both groups were significantly lower than those before treatment(P<0.05),while the d-MDBP in the treatment group after treatment was slightly higher than that in the control group(P<0.05).The score of the Du's Hypertension Quality of Life Scale in the treatment group after treatment was higher than that before treatment(P<0.05),the score in the treatment group was higher than that in the control group,but the difference was not statistically significant(P>0.05).Conclusion:Acupuncture at Neiguan(PC6)-Jueyinshu(BL14)combined with oral western medicine had a good regulatory effect on the circadian rhythm of blood pressure in patients with non-dipper hypertension,and its effect was superior to that of oral western medicine alone.Both acupuncture at Neiguan(PC6)-Jueyinshu(BL14)combined with oral western medicine and oral western medicine alone had a certain hypotensive effect on patients with non-dipper hypertension.In terms of nocturnal mean systolic blood pressure,the hypotensive effect of combined acupuncture and medication was better than that of oral western medicine alone.In addition,acupuncture at Neiguan(PC6)-Jueyinshu(BL14)combined with oral western medicine could effectively improve the clinical accompanying symptoms and quality of life in patients with non-dipper hypertension.

Abstract:Objective:To evaluate the methodological and reporting quality of systematic reviews/Meta-analyses on traditional Chinese medicine for ischemic cardiomyopathy(ICM).Methods:Two researchers independently searched the China National Knowledge Infrastructure(CNKI),the Chinese Biomedical Literature Database(CBM),WanFang Data,the Chinese Science and Technology Journal Database(VIP),PubMed,and the Medscape FMRS foreign medical information platform.Chinese and English literature on systematic reviews/Meta-analyses of traditional Chinese medicine for treatment of ICM were collected from the establishment of database to October 31,2023.The AMSTAR scale and the PRISMA 2020 scale were used to conduct methodological and reporting quality evaluations respectively.Results:A total of 12 systematic reviews/Meta-analyses literatures were included,involving 10 literatures in Chinese and 2 literatures in English.The average score of the AMSTAR scale was(7.71±1.42)with the scores for Chinese and English literatures being(7.30±1.16)and(9.75±0.35)respectively.In the evaluation results of the AMSTAR scale,two English literatures registered the prospective research protocol and clearly stated the relevant conflicts of interest.In the AMSTAR scale evaluation results,two English literatures registered the prospective study protocol and clearly stated the relevant conflicts of interest.The items with lower scores in the Chinese literature mainly included whether to provide the preliminary design plan,whether to provide a list of research literature for inclusion and exclusion,and whether to make a conflict-of-interest statement.None of the literatures provided a list of excluded evaluation literature.None of the 10 included Chinese literatures were high quality,with the low or medium quality category.Two English literatures were high quality.The average PRISMA 2020 score was(19.92±3.42),while the scores for the Chinese and English literatures were(18.95±2.82)and(24.75±1.06)respectively.The Chinese literatures scored lower primarily on items related to search strategies,credibility evaluation,registration and protocol,conflict of interest statements,and the availability of other materials.Conclusion:Systematic reviews/Meta-analyses on traditional Chinese medicine for ICM exhibited issues such as lacking forward-looking research design schemes,without related conflicts of interest,list of excluded studies,and insufficient result.The quality of Chinese literatures was generally lower than that of English literatures.Systematic reviews/Meta-analyses on traditional Chinese medicine for ICM should comply with the requirements of AMSTAR and PRISMA 2020 scales,and form high-quality systematic reviews/Meta-analyses to provide high-quality evidence for clinical practice.

Abstract:Objective:To explore the mechanism of Rhodiola Crenulata for treating plateau hypoxic brain injury based on reverse molecular docking and experimental verification.Methods:The chemical components of Rhodiola Crenulata were collected by searching the comprehensive Chinese medicine database(ETCM),the database of the association between Chinese medicine and disease symptoms(SymMap),and through supplementary literature collection.The active ingredients with absorption,distribution,metabolism,and excretion(ADME)prediction results were screened using the drug screening platform(FAFDrugs4).The potential targets of active ingredients were predicted by reverse molecular docking method.The targets related to plateau hypoxic brain injury were obtained from the human gene database(GeneCards).The common targets of medicinal materials and diseases were obtained.The"traditional Chinese medicine-component-target-disease"regulatory network and the common target protein-protein interaction(PPI)network were constructed.The core targets were obtained through network topology analysis.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were conducted using the OmicShare software,and in vivo pharmacodynamic experiments were carried out for verification.The regulatory effect of Rhodiola Crenulata on the expression of core target messenger ribonucleic acid(mRNA)in the hippocampal tissue of rats with plateau hypoxic brain injury was detected by the real-time fluorescent quantitative polymerase chain reaction(RT-PCR).Western blot was used to detect the regulatory effect of Rhodiola Crenulata on the protein expression of serine/threonine protein kinase 1(AKT1)and signal transducer and activator of transcription 3(STAT3)in brain tissue of rats with low-pressure oxygen combined with lipopolysaccharide(LPS)-induced brain injury,further confirming the results of network pharmacology analysis.Results:A total of 76 active components and 285 targets were screened from Rhodiola Crenulata.One hundred and forty-eight disease targets related to plateau hypoxic brain injury were obtained,with 23 common targets.The core targets included AKT1,heat shock protein 90α1(HSP90AA1),albumin(ALB),estrogen receptor 1(ESR1),and STAT3.GO enrichment analysis yielded 1 607 entries,and KEGG enrichment analysis identified 71 pathways.Rhodiola Crenulata could down-regulate the mRNA expression levels of the core targets serine/AKT1 and STAT3.Rhodiola Crenulata could inhibit the phosphorylation of serine/AKT1 protein,and all doses could significantly inhibit the expression of STAT3 protein(P＜0.01).Conclusion:Rhodiola Crenulata could treat plateau hypoxic brain injury through multi-target and multi-pathway.

Abstract:Objective:To explore the mechanism of the Tiaogan Formula in preventing and treating coronary heart disease with Qi stagnation and blood stasis syndrome.Methods:Based on the"disease-syndrome-formula"core concept,network analysis,bioinformatics and molecular docking verification,the effective material basis were systematically screened through the databases of traditional Chinese medicine syndromes,traditional Chinese medicine and diseases,and the drug targets were identified.Results:Tiaogan Formula mainly consisted of Xuefu Zhuyu Decoction,Chaihu Shugan Powder and Xiao Yao Powder.A total of 357 targets for Tiaogan Formula were identified through the TCMIP v2.0 platform.By the DisGeNET,GeneCards,OMIM,PharmGKB,TTD disease databases,and SymMap v2 syndrome database,424 targets for coronary heart disease with Qi stagnation and blood stasis syndrome were obtained.A target network of Tiaogan Formula in preventing and treating coronary heart disease with Qi stagnation and blood stasis syndrome was constructed,including 9 core targets,namely tumor necrosis factor(TNF),DNA methyltransferase 1(DNMT1),phosphatidylinositol 3-kinase catalytic subunit alpha(PIK3CA),prostaglandin endoperoxide synthase 2(PTGS2),estrogen receptor beta(ESR2),insulin(INS),caspase-3(CASP3),phosphatidylinositol 3-kinase regulatory subunit 1(PIK3R1),and peroxisome proliferator-activated receptor gamma(PPARG).A total of 6 core chemical components were obtaind by the"chemical composition-target"network combined with the comprehensive evaluation of drugability,including Formononetin,3,3'-Dimethylquercetin,Pinocembrin,Kumatakenin,3',7-Dihydroxy-4',6-Dimethoxyisoflavone,and Liquiritigenin.Molecular docking was used to verify that there was a strong interaction between the core targets and the core chemical components.Gene ontology(GO)functional enrichment analysis yielded 71 biological processes(BP),5 cellular components(CC),and 13 molecular functions(MF).Kyoto encyclopedia of genes and genomes(KEGG)signaling pathway enrichment analysis obtained 72 items,and the significantly enriched signaling pathways included lipid and atherosclerosis(hsa05417),vascular endothelial growth factor(VEGF,hsa04370),TNF(hsa04668),regulation of adipocyte lipolysis(hsa04923),adenosine monophosphate-activated protein kinase(AMPK,hsa04152),and cell apoptosis(hsa04210),mainly involving mechanisms such as vascular physiology,cell death,glycolipid metabolism,energy metabolism,and immune inflammation.Conclusion:Tiaogan Formula in preventing and treating coronary heart disease with Qi stagnation and blood stasis syndrome was closely related to the mechanisms of vascular remodeling,regulating cell death,regulating glucose and lipid metabolism,energy metabolism,and immune inflammation.

Abstract:Objective:To investigate the protective effect of Qijing Huixin Formula on the heart based on network pharmacology and basic study.Methods:The potential active components of Qijing Huixin Formula and its corresponding target sites were screened out through the drug target prediction website and by reviewing the literature.Using the disease database,the related target sites of acute myocardial infarction(AMI)were retrieved,and the intersection of the two target sites was obtained.The interaction network diagram of drug active components and target sites,the protein-protein interaction(PPI)network diagram were drawn.The gene ontology(GO)functional enrichment analysis,Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were conducted,and molecular docking was used for verification.The therapeutic effect and mechanism of Qijing Huixin Formula in treating AMI rats were verified.Results:Qijing Huixin Formula for treating AMI involved 239 drug targets.The quercetin,kaempferol,digitonin flavonoids,β-glucoside,and Tanshinone ⅡA showed the greatest effect.Protein kinase B1(AKT1),glycogen synthase kinase 3β(GSK3β),vascular endothelial growth factor A(VEGFA),tumor necrosis factor(TNF),and interleukin 6(IL6)were the key targets.The main enriched pathways included the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway,TNF signaling pathway,and apoptosis signaling pathway.The cardiac function of AMI rats,the levels of serum myocardial enzymes,lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),and inflammatory factors(TNF-α,IL-6),microvascular density,the mRNA expressions of VEGFA and the markers of the PI3K/AKT/catenin β1(CTNNB1)pathway in the myocardial infarction edge zone improved by treatment of Qijing Huixin Formula(P＜0.05).Conclusion:Qijing Huixin Formula could improve myocardial injury in AMI rats by activating the PI3K/GSK3β/CTNNB1 signaling pathway in myocardial tissue and promoting angiogenesis,thereby enhancing cardiac function.

Abstract:Objective:To explore the effect of Tianxiang Dan in inhibiting the progression of atherosclerosis in mice by regulating the autophagy-mammalian target of rapamycin(mTOR)-absent in melanoma 2(AIM2)inflammatoryosome axis and its mechanism.Methods:Atherosclerosis models of ApoE knockout(ApoE-/-)mice were established by feeding with a high-fat diet for 12 weeks.The mice were divided into the model group(MD group),Tianxian Dan group[2.73 g/(kg·d),TXD group],ApoE-/-·NACHT,LRR and PYD domain protein 3(NLRP3)-/-+TXD group[2.73 g/(kg·d),NLRP3-/-group]and ApoE-/-·NLRP3-/-+RGFP966 group[400 μg/(kg·d),RGFP966 group],with 10 mice in each group.Ten C57BL/6J mice were fed with the basic diet as the blank control group(CO group).The plaque burden and stability were evaluated by oil red O staining,hematoxylin-eosin(HE)and Masson staining.The morphology of autophagosomes in aortic tissue was observed under transmission electron microscopy.The expressions of microtubule-associated protein 1 light chain 3(LC3),myosin-like BCL2-binding protein(Beclin-1),phosphorylated mTOR(p-mTOR),AIM2,Caspase-1,pyroptosis factor D(GSDMD),interleukin-1β(IL-1β),and interleukin-18(IL-18)proteins were detected by Western Blot.The mRNA expression of AIM2 was detected by immunohistochemistry and real-time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR).The levels of IL-1β and IL-18 in serum and aortic homogenate were determined by enzyme-linked immunosorbent assay(ELISA),and correlation analysis was performed.Results:Compared with the MD group,the protein expressions of AIM2,p-mTOR,Caspase-1,GSDMD,IL-1β,IL-18,and the serum levels of IL-1β and IL-18,the mRNA expression AIM2 in the TXD group decreased,while the protein expressions of LC3 and Beclin-1 increased(P<0.05).Compared with the TXD group,the protein expressions of AIM2,p-mTOR,Caspase-1,GSDMD,and AIM2 mRNA in the NLRP3-/-group increased,while the protein expressions of IL-18,LC3,Beclin-1,and the serum content of IL-18 decreased(P<0.05).Compared with the TXD group,the protein expression and serum content of IL-18 in the RGFP966 group decreased(P<0.05).Correlation analysis showed that the autophagy level was negatively correlated with AIM2,GSDMD,IL-1β,IL-18,and p-mTOR were LC3 correlated with AIM2,GSDMD,IL-1β,and IL-18.Conclusion:Tianxiang Dan could reduce plaque burden and enhance plaque stability and its mechanism might be related to the regulation of the autophagy-mTOR-AIM2 inflammatory pathway.

Abstract:Objective:To observe the effect of overexpression of collagen triple helix repeat containing-1(CTHRC1)in vitro on pathological myocardial remodeling induced by angiotensin Ⅱ(Ang Ⅱ)and the related mechanisms.Methods:The rat H9c2 cardiomyocytes cultured in vitro were divided into the GFP group(transfected with negative control GFP-AAV9 type gene adenovirus 4.8×1010 PFU/mL)and the CTHRC1 group(transfected with CTHRC1-AAV9 type gene adenovirus 2.4×1010 PFU/mL).The GFP group was further divided into the Vehicle-GFP group(transfected with negative control GFP-AAV9 type gene adenovirus 4.8×1010 PFU/mL)and the GFP-AngⅡ group(transfected with negative control GFP-AAV9 type gene adenovirus 4.8×1010 PFU/mL,AngⅡ 10 μmol/L).the CTHRC1 group was divided into the Vehicle-CTHRC1 group(transfected with CTHRC1-AAV9 type gene adenovirus 2.4×1010 PFU/mL)and the CTHRC1-AngⅡ group(transfected with CTHRC1-AAV9 type gene adenovirus 2.4×1010 PFU/mL,AngⅡ 10 μmol/L).The expression of CTHRC1 protein was detected by Western Blot.The surface area of H9c2 cells was evaluated by phalloidin staining.After 10％sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),the gel was stained with Coomassie Brilliant blue,cut,and protein identification was performed using high-resolution mass spectrometry.Gene Ontology(GO)gene,and set enrichment analysis(GSEA)functional enrichment analysis was conducted for the differentially expressed genes.The activities of superoxide dismutase(SOD),catalase(CAT),and malondialdehyde(MDA)were detected using kits.Results:Compared with the Vehicle-GFP group,the surface area of H9c2 cells in the GFP-AngⅡ group increased(P<0.001).Compared with the Vehicle-CTHRC1 group,the surface area of H9c2 cells in the CTHRC1-AngⅡgroup increased(P<0.001).Compared with the GFP-AngⅡ group,the surface area of H9c2 cells in the CTHRC1-AngⅡ group increased(P<0.001).Compared with the Vehicle group,the expression of H9c2 cardiomyocytes in the AngⅡ group decreased around 75 kDa.Compared with the GFP group,the CTHRC1 group showed 31 proteins significantly downregulated and 30 proteins significantly upregulated(P<0.05).For the significantly downregulated proteins,the GO analysis showed that signaling pathways such as glutathione metabolism and protein stability were significantly enriched.For the significantly downregulated proteins,the GSEA analysis showed that the glutathione metabolism pathway was significantly enriched.Compared with the GFP-AngⅡ group,the H9c2 cardiomyocytes in the Vehicle-GFP group showed lower SOD and CAT activities and higher MDA activity(P<0.05 or P<0.001).Compared with the Vehicle-CTHRC1 group,the H9c2 cardiomyocytes in the CTHRC1-AngⅡ group showed lower SOD and CAT activities and higher MDA activity(P<0.01 or P<0.001).Compared with the GFP-AngⅡ group,the H9c2 cardiomyocytes in the CTHRC1-AngⅡ group showed lower SOD and CAT activities and higher MDA activity(P<0.05 or P<0.001).Conclusion:Overexpression of CTHRC1 in vitro could aggravate pathological myocardial remodeling induced by AngⅡ,and its function was related to the inhibition of the glutathione metabolism pathway.