ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

Based on LI Gao's Academic Thought， focusing on the process of qi transformation and taking the regulation and restoration of metabolism and immunity as the entry point， a "source-pivot-convergence" diagnostic and therapeutic model for malignant tumors is constructed. In this model， spleen and stomach internal injury is the source of malignant tumor occurrence， while the disorder of ascending and descending is the pivot of the disease development， and the generation of yin fire is the convergence of malignant tumor progression. Based on this， the three major therapeutic methods of clearing the source， harmonizing the pivot， and resolving the convergence are established. To fortify spleen and boost qi， consolidate the root and clear the source， modified Buzhong Yiqi Decoction（补中益气汤）can be used. To raise the clear and direct the turbid downward， regulate qi and harmonize the pivot， modified Shengyang Yiwei Decoction （升阳益胃汤） is suggested. To restore balance and promote circulation， disperse accumulation and resolve convergence， modified Shengyang Sanhuo Decoction （升阳散火汤） is selected. In clinical practice， these formulas can be used in combination according to the complexity of the pathogenesis， and further adapted with prescriptions for promoting dispersion and penetrating pathogenic factors， resolving phlegm and promoting circulation， activating blood and eliminating concretions， which can provide a reference for the prevention and treatment of tumor diseases.

Based on LI Gao's Academic Thought， focusing on the process of qi transformation and taking the regulation and restoration of metabolism and immunity as the entry point， a "source-pivot-convergence" diagnostic and therapeutic model for malignant tumors is constructed. In this model， spleen and stomach internal injury is the source of malignant tumor occurrence， while the disorder of ascending and descending is the pivot of the disease development， and the generation of yin fire is the convergence of malignant tumor progression. Based on this， the three major therapeutic methods of clearing the source， harmonizing the pivot， and resolving the convergence are established. To fortify spleen and boost qi， consolidate the root and clear the source， modified Buzhong Yiqi Decoction（补中益气汤）can be used. To raise the clear and direct the turbid downward， regulate qi and harmonize the pivot， modified Shengyang Yiwei Decoction （升阳益胃汤） is suggested. To restore balance and promote circulation， disperse accumulation and resolve convergence， modified Shengyang Sanhuo Decoction （升阳散火汤） is selected. In clinical practice， these formulas can be used in combination according to the complexity of the pathogenesis， and further adapted with prescriptions for promoting dispersion and penetrating pathogenic factors， resolving phlegm and promoting circulation， activating blood and eliminating concretions， which can provide a reference for the prevention and treatment of tumor diseases.

Objective To investigate the current status of hypertension in the old adults living in urban city and rural areas in Qinghai Plateau and analyze the related influencing factors in order to provide data and evidence for targeted formulation of preventive and control measures for the pop-ulation.Methods Cluster-random sampling was used to subject 1372 elderly people(aged ≥60 years)from 8 urban areas and 25 natural villages in Xining City,Qinghai Province.Questionnaires were used to collect their demographic data,body mass index(BMI),history of chronic diseases,and lipid-related indicators.According to complicated with hypertension or not,they were divided into a hypertension group(615 cases)and a non-hypertension group(757 cases).SPSS 26.0 soft-ware was employed to perform statistical analyses with descriptive analysis and multivarlate un-conditional logistic regression analysis.Results Among the 1372 elderly persons,615 participants had hypertension,and the overall prevalence was 44.8％,and that in urban area and rural area was 50.1％and 38.5％,respectively,with significant difference(P＜0.01).Statistical differences were observed between those with and without hypertension in terms of age,BMI,and proportions of coronary heart disease(CHD),diabetes and stroke(P＜0.05,P＜0.01).In the urban populations,there were obvious differences in marital status,BMI,and proportions of CHD and diabetes be-tween those with and without hypertension(P＜0.01).For the rural populations,notable differ-ences were observed in age and proportions of CHD and diabetes between those with and without hypertension(P＜0.05,P＜0.01).Multivariate unconditional logistic regression analysis revealed that urban areas,obesity,CHD and diabetes were risk factors for hypertension in the elderly living in the urban and rural areas(OR=1.622,95％CI:1.299-2.026,P=0.000;OR=0.564,95％CI:0.315-1.006,P=0.042;OR=0.604,95％CI:0.417-0.874,P=0.008;OR=0.472,95％CI:0.328-0.678,P=0.000;OR=0.474,95％CI:0.334-0.673,P=0.000).Obesity,CHD and diabetes were risk factors for hypertension in those in the urban areas(OR=0.553,95％CI:0.317-0.963,P=0.036;OR=0.506,95％CI:0.320-0.800,P=0.004;OR=0.458,95％CI:0.303-0.692,P=0.000),and CHD and diabetes were risk factors in those in the rural areas(OR=0.382,95％CI:0.219-0.666,P=0.001;OR=0.452,95％CI:0.253-0.807,P=0.007).Conclusion There is sig-nificant difference in the prevalence of hypertension between the elderly living in the urban city and rural areas in Qinghai Plateau.The old adults with overweight,obesity,and complication of CHD and diabetes are prone to developing hypertension.

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Objective: To explore the mechanism of action of Zangjiangzhi capsule (ZJZC) in treating hyperlipidemia (HLP). Methods: The components of ZJZC were analyzed and identified using ultra-high performance liquid chromatography with Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-Exactive-Orbitrap-MS/MS). Network pharmacology analysis was used to explore the mechanism of action of ZJZC in HLP treatment. The SwissTargetPrediction database was used to predict compound targets, and GeneCards, DisGeNet, OMIM, and DRUGBANK databases were used to identify HLP-related targets. Protein–protein interaction diagrams were constructed using the STRING database. The targets were subjected to gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The “herb-ingredient-target” network was visualized using Cytoscape. Preliminary validation was performed using molecular docking and enzyme-linked immunosorbent assay. Results: Ninety compounds were identified in ZJZC, including 34 flavonoids, 12 phenols, 10 terpenoids, 10 alkaloids, 8 organic acids, 8 anthraquinones, and 9 other compounds. In total, 904 targets were identified for these compounds. Among them, 158 targets intersected with the HLP target network. Network pharmacology analysis showed that MAPK1, PPAR-α, RXRA, HSP90AA1, PIK3R1, AKT1, PIK3CA, IL6, TNF, and ESR1 are the key targets of action. KEGG enrichment analysis identified 164 pathways. Among these, the AGE-RAGE signaling pathway in diabetic complications, lipid and atherosclerosis pathways, regulation of lipids in adipocytes, and insulin resistance are related to HLP. Molecular docking showed good affinity between the key targets and ingredients. Further, ZJZC treatment in mice resulted in lower expression of MAPK1 protein and increased expression of PPAR-α protein, which have been shown to be strongly associated with HLP. Conclusions This study showed that ZJZC contains various active ingredients and can modulate multiple targets and pathways associated with HLP, providing evidence at the molecular level for its clinical application in the treatment of HLP.

Objective To assess the efficacy and safety of Tiaozhou Ziyin(TZZY)recipe for treatment of premature ovarian insufficiency(POI)and explore the possible mechanisms.Methods We used bioinformatics analyses and network pharmacology to identify the main active ingredients in TZZY recipe and their core targets,which were verified by Western blotting.We tested the efficacy and safety of the recipe in 60 POI patients,who were randomized into control group(n=30)with Femoston treatment and TZZY group(n=30)with additional TZZY recipe treatment for 3 menstrual cycles.Results The core active ingredients of TZZY recipe included kaempferol,β-sitosterol,luteolin,and quercetin.The core targets included SRC,TP53,STAT3,PIK3CA,and MAPK3,which were involved in positive regulation of cell movement and protein phosphorylation,the cancer pathways and the PI3K-Akt signaling pathway.Molecular docking showed that the core active ingredients had good binding ability with the core targets.In female rat models of POI,TZZY recipe treatment significantly up-regulated ovarian expressions of p-PI3K and p-Akt proteins.In the clinical trial,treatment with Femoston and Femoston plus TZZY recipe both significantly increased E2 levels and reduced FSH and LH levels and Kupperman scores of the patients,and the combined treatment produced significantly stronger effects.Both treatments increased the number of antral follicles of the patients,but the combined treatment also significantly increased the levels of AMH.Conclusion The therapeutic mechanism of TZZY recipe for POI involves multiple active ingredients,multiple therapeutic targets and multiple pathways,and activating the PI3K/Akt pathway is one of its main mechanisms of action,to improve ovarian reserve function,alleviate clinical symptoms,and enhance clinical efficacy in POI patients.

OBJECTIVES: To assess the efficacy and safety of Tiaozhou Ziyin (TZZY) recipe for treatment of premature ovarian insufficiency (POI) and explore the possible mechanisms. METHODS: We used bioinformatics analyses and network pharmacology to identify the main active ingredients in TZZY recipe and their core targets, which were verified by Western blotting. We tested the efficacy and safety of the recipe in 60 POI patients, who were randomized into control group (n=30) with Femoston treatment and TZZY group (n=30) with additional TZZY recipe treatment for 3 menstrual cycles. RESULTS: The core active ingredients of TZZY recipe included kaempferol, β-sitosterol, luteolin, and quercetin. The core targets included SRC, TP53, STAT3, PIK3CA, and MAPK3, which were involved in positive regulation of cell movement and protein phosphorylation, the cancer pathways and the PI3K-Akt signaling pathway. Molecular docking showed that the core active ingredients had good binding ability with the core targets. In female rat models of POI, TZZY recipe treatment significantly up-regulated ovarian expressions of p-PI3K and p-Akt proteins. In the clinical trial, treatment with Femoston and Femoston plus TZZY recipe both significantly increased E₂ levels and reduced FSH and LH levels and Kupperman scores of the patients, and the combined treatment produced significantly stronger effects. Both treatments increased the number of antral follicles of the patients, but the combined treatment also significantly increased the levels of AMH. CONCLUSIONS The therapeutic mechanism of TZZY recipe for POI involves multiple active ingredients, multiple therapeutic targets and multiple pathways, and activating the PI3K /Akt pathway is one of its main mechanisms of action, to improve ovarian reserve function, alleviate clinical symptoms, and enhance clinical efficacy in POI patients.
Female ; Primary Ovarian Insufficiency/drug therapy* ; Humans ; Drugs, Chinese Herbal/therapeutic use* ; Animals ; Rats ; Molecular Docking Simulation ; Signal Transduction ; Sitosterols/therapeutic use* ; Kaempferols/therapeutic use*

Objective To investigate the current status of hypertension in the old adults living in urban city and rural areas in Qinghai Plateau and analyze the related influencing factors in order to provide data and evidence for targeted formulation of preventive and control measures for the pop-ulation.Methods Cluster-random sampling was used to subject 1372 elderly people(aged ≥60 years)from 8 urban areas and 25 natural villages in Xining City,Qinghai Province.Questionnaires were used to collect their demographic data,body mass index(BMI),history of chronic diseases,and lipid-related indicators.According to complicated with hypertension or not,they were divided into a hypertension group(615 cases)and a non-hypertension group(757 cases).SPSS 26.0 soft-ware was employed to perform statistical analyses with descriptive analysis and multivarlate un-conditional logistic regression analysis.Results Among the 1372 elderly persons,615 participants had hypertension,and the overall prevalence was 44.8％,and that in urban area and rural area was 50.1％and 38.5％,respectively,with significant difference(P＜0.01).Statistical differences were observed between those with and without hypertension in terms of age,BMI,and proportions of coronary heart disease(CHD),diabetes and stroke(P＜0.05,P＜0.01).In the urban populations,there were obvious differences in marital status,BMI,and proportions of CHD and diabetes be-tween those with and without hypertension(P＜0.01).For the rural populations,notable differ-ences were observed in age and proportions of CHD and diabetes between those with and without hypertension(P＜0.05,P＜0.01).Multivariate unconditional logistic regression analysis revealed that urban areas,obesity,CHD and diabetes were risk factors for hypertension in the elderly living in the urban and rural areas(OR=1.622,95％CI:1.299-2.026,P=0.000;OR=0.564,95％CI:0.315-1.006,P=0.042;OR=0.604,95％CI:0.417-0.874,P=0.008;OR=0.472,95％CI:0.328-0.678,P=0.000;OR=0.474,95％CI:0.334-0.673,P=0.000).Obesity,CHD and diabetes were risk factors for hypertension in those in the urban areas(OR=0.553,95％CI:0.317-0.963,P=0.036;OR=0.506,95％CI:0.320-0.800,P=0.004;OR=0.458,95％CI:0.303-0.692,P=0.000),and CHD and diabetes were risk factors in those in the rural areas(OR=0.382,95％CI:0.219-0.666,P=0.001;OR=0.452,95％CI:0.253-0.807,P=0.007).Conclusion There is sig-nificant difference in the prevalence of hypertension between the elderly living in the urban city and rural areas in Qinghai Plateau.The old adults with overweight,obesity,and complication of CHD and diabetes are prone to developing hypertension.

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.