ObjectiveThis study aims to elucidate the potential mechanism of Zuojinwan in improving liver fibrosis through hepatic tissue metabolomics analysis. MethodsTwenty-four mice were randomly allocated into normal group， model group ， positive drug group （silymarin， 100 mg·kg^-1）， and Zuojinwan group （Zuojinwan solution， 2.5 g·kg^-1）， with per group six mice. Liver fibrosis model was induced via intraperitoneal injection of olive oil solution with 10% carbon tetrachloride （CCl₄） （0.5 μL·g^-1， three times weekly for 8 weeks） in all groups except the normal group. During the final 4 weeks， the silymarin group received silymarin （100 mg·kg^-1） by gavage thrice weekly， while the Zuojinwan group was administered Zuojinwan solution （2.5 g·kg^-1） under the same regimen. After the last administration， the levels of liver fibrosis indicators and liver injury markers in serum were detected. The pathological morphological changes of the liver tissues were observed. The levels of liver fibrosis markers α-smooth muscle actin （α-SMA） and Collagen Ⅰ（ColⅠ） were detected. Metabolomics was analyzed on mice's liver tissues. The mice's serum was collected for metabolomics analysis. ResultsCompared with the model group， Zuojinwan significantly improved indicators related to liver fibrosis and liver injury. Compared with the normal group， the model group showed significantly elevated levels of fibrosis markers such as laminin （LN）， hyaluronic acid （HA）， procollagen typeⅢ （PC-Ⅲ）， and type Ⅳ Collagen （Ⅳ-C）， while liver injury indicators such as alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， alkaline phosphatase （ALP）， lactate dehydrogenase （LDH）， and total bilirubin （TBIL）， exhibited a marked upward trend （P<0.05）. Compared with the model group， the silymarin group showed a significant decrease in the aforementioned indicators （P<0.05）. Notably， compared with the model group， the Zuojinwan group exhibited a significant reduction in all these indicators （P<0.05）， with efficacy comparable to that of the silymarin group. Zuojinwan reduced mRNA and protein levels of α-SMA and ColⅠ in the liver tissue. Metabolomics results revealed that compared with the model group， Zuojiinwan significantly reduced levels of glucose metabolism-related metabolites such as D-fructose 1，6-bisphosphate （FBP）， nicotinamide adenine dinucleotide phosphate （NADPH）， sodium beta-D-fructose 6-phosphate （F6P）， dihydroxyacetone phosphate （DHAP）， fumaric acid， and D-glucose 6-phosphate （G6P） （P<0.05）. Serum enzyme-linked immunosorbent assay （ELISA） was used to detect glucose metabolism indicators and further validate the regulatory effect of Zuojinwan on glucose metabolism. ConclusionThese results suggest that Zuojinwan may improve liver fibrosis by regulating the dysregulated levels of glucose metabolism during the progression of liver fibrosis.

BACKGROUND:The stress effect of autophagy on epithelial cells,fibroblasts and myofibroblasts is closely related to the formation process of pulmonary fibrosis.OBJECTIVE:To screen the genes related to autophagy in patients with pulmonary fibrosis,and explore their correlation with the prognosis of patients with pulmonary fibrosis,in order to provide a new target for clinical intervention in pulmonary fibrosis.METHODS:The gene expression profiling dataset downloaded from GSE70866 was used as a training set,differentially expressed genes between pulmonary fibrosis patients and normal healthy individuals was analyzed using the R language and intersected with autophagy-related genes to identify the differentially expressed genes with the most significant changes.Multiple analysis methods were used to identify key prognostic genes and construct genetic prognostic models.Patients with pulmonary fibrosis were divided into high-risk and low-risk groups according to their risk scores,and the validity of the prognostic model was verified using the Siena cohort and Leuven cohort validation sets.A cell model of pulmonary fibrosis was established by inducing HFL-1 cells(human embryonic lung fibroblasts)with transforming growth factor-β1,and an animal model of pulmonary fibrosis was established in mice by tracheal instillation of bleomycin to validate the expressions of prognostic genes.RESULTS AND CONCLUSION:(1)There were 2 650 differentially expressed genes between fibrotic tissue and normal tissue.Among them,34 genes related to autophagy showed significant expression changes.(2)Kaplan-Meier survival analysis curves for the Siena cohort and Leuven cohort validation sets showed significantly lower survival in the high-risk group than in the low-risk group.(3)Three autophagy genes related to prognosis were screened out:myelocytomatosis viral oncogene(MYC),C-C motif chemokine ligand 2(CCL2),and GABA type a receptor associated protein like 1(GABARAPL1).(4)Both in vivo and in vitro studies showed that compared with the control group,the expression levels of myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 mRNA and protein were significantly higher in the lung fibrosis model group(P＜0.01,P＜0.05),while the expression levels of GABA type a receptor associated protein like 1 mRNA and protein were lower(P＜0.001).To conclude,bioinformatics methods are used to analyze the expression of three autophagy-related genes in pulmonary fibrosis and their correlation with the prognosis of patients with pulmonary fibrosis.The constructed prognostic model has good predictive ability for the 1-,2-,and 3-year survival rates of patients with pulmonary fibrosis.Moreover,in vivo and in vitro models have been used to verify that myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 are highly expressed in lung fibroblasts and tissues,and that GABA type a receptor associated protein like 1 is lowly expressed.

Over the past two decades, genome-wide association study(GWAS) has identified numerous genetic variants and loci associated with heritable diseases. With the gradual maturation and saturation of GWAS methodologies, transcriptome-wide association study(TWAS) offers a novel perspective by linkinggenetic phenotypes to gene expression levels. By integrating TWAS with other multi-omics analyses, researchers can gain a deeper understanding of heritable diseases. This article provides an overview of recent groundbreaking and representative TWAS methods and tools, analyzes their strengths and limitations, and discusses future trends in TWAS development.

Based on the theory that Shaoyang （少阳） and Shaoyin （少阴） function as the principal pivots， and integrating the laws of qi-fire transformation， yin-yang alternation， and the waxing and waning of healthy qi and pathogenic factors， as well as the action characteristics of immune checkpoint inhibitors （ICIs）， this study systematically expounds the dynamic evolution laws of the Shaoyang and Shaoyin pivots and constructs a stage- and phase-based differentiation and treatment framework for gastric cancer immunotherapy. In the neoadjuvant treatment stage， the core pathogenesis is the dysfunction of the Shaoyang pivot and the disorder of qi and fire transformation， often accompanied by stagnation of cold-deficiency in the middle jiao （焦）. In the postoperative adjuvant treatment stage， the dominant factors are the depletion of Shaoyin fire and the damage of yin fluid， accompanied by the lingering of residual pathogen in the Shaoyang level and the obstruction of collaterals by phlegm and stasis. In the advanced stage， the critical conditions are the exhaustion of the Shaoyin pivot， the blockage of orifices by turbid toxins， and the separation of yin and yang， leading to the rampant of pathogenic toxins and the emergence of various complicated syndromes. The treatment should be guided by the principle of regulating the double pivot. In the neoadjuvant treatment stage， it is recommended to activate the Shaoyang pivot and warm and promote the qi movement in the middle jiao. In the postoperative adjuvant treatment stage， clearing the residual pathogenic factors from the pivot and nourishing the Shaoyin. In the advanced stage， rescuing the Shaoyin pivot and opening the orifices to transform turbidity. Furthermore， stage-based treatment should serve as the foundation for dynamically assessing the patient's immune status and phase-specific changes in immunity， thereby promoting immune activation while preventing and managing immune overactivation and drug resistance.

BACKGROUND:The stress effect of autophagy on epithelial cells,fibroblasts and myofibroblasts is closely related to the formation process of pulmonary fibrosis.OBJECTIVE:To screen the genes related to autophagy in patients with pulmonary fibrosis,and explore their correlation with the prognosis of patients with pulmonary fibrosis,in order to provide a new target for clinical intervention in pulmonary fibrosis.METHODS:The gene expression profiling dataset downloaded from GSE70866 was used as a training set,differentially expressed genes between pulmonary fibrosis patients and normal healthy individuals was analyzed using the R language and intersected with autophagy-related genes to identify the differentially expressed genes with the most significant changes.Multiple analysis methods were used to identify key prognostic genes and construct genetic prognostic models.Patients with pulmonary fibrosis were divided into high-risk and low-risk groups according to their risk scores,and the validity of the prognostic model was verified using the Siena cohort and Leuven cohort validation sets.A cell model of pulmonary fibrosis was established by inducing HFL-1 cells(human embryonic lung fibroblasts)with transforming growth factor-β1,and an animal model of pulmonary fibrosis was established in mice by tracheal instillation of bleomycin to validate the expressions of prognostic genes.RESULTS AND CONCLUSION:(1)There were 2 650 differentially expressed genes between fibrotic tissue and normal tissue.Among them,34 genes related to autophagy showed significant expression changes.(2)Kaplan-Meier survival analysis curves for the Siena cohort and Leuven cohort validation sets showed significantly lower survival in the high-risk group than in the low-risk group.(3)Three autophagy genes related to prognosis were screened out:myelocytomatosis viral oncogene(MYC),C-C motif chemokine ligand 2(CCL2),and GABA type a receptor associated protein like 1(GABARAPL1).(4)Both in vivo and in vitro studies showed that compared with the control group,the expression levels of myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 mRNA and protein were significantly higher in the lung fibrosis model group(P＜0.01,P＜0.05),while the expression levels of GABA type a receptor associated protein like 1 mRNA and protein were lower(P＜0.001).To conclude,bioinformatics methods are used to analyze the expression of three autophagy-related genes in pulmonary fibrosis and their correlation with the prognosis of patients with pulmonary fibrosis.The constructed prognostic model has good predictive ability for the 1-,2-,and 3-year survival rates of patients with pulmonary fibrosis.Moreover,in vivo and in vitro models have been used to verify that myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 are highly expressed in lung fibroblasts and tissues,and that GABA type a receptor associated protein like 1 is lowly expressed.

OBJECTIVE To investigate the equivalence of different decoction processes based on rat model of hypertension with liver-yang hyperactivity. METHODS Inductively coupled plasma mass spectrometry （ICP-MS） was used to compare the dissolution differences of inorganic elements in the powder-directly-decocted decoction versus the pieces-decocted-first decoction of Ostreae Concha- Haliotidis Concha- Margaritifera Concha drug pair. Six SD rats were included in the normal group. The spontaneously hypertensive rats were given Aconite decoction for six weeks to induce the hypertension model with liver-yang hyperactivity. After successful modeling， 48 rats were randomly divided into the model group， the captopril group [positive control， 8 mL/（kg·d） ] ， as well as low-， medium-， and high-dose groups of pieces decocted first or directly powder decocted [2.02， 4.05， 8.10 mL/（kg·d） ] ， with 6 rats in each group. Each group received the corresponding drug or equal volume of pure water intragastrically， once a day， for two consecutive weeks. After the last administration， the degree of irritability， facial temperature， pressure pain threshold， blood pressure， and pathological changes of the thoracic aorta were observed in each group. Serum nitric oxide （NO） and plasma angiotensin Ⅱ （Ang Ⅱ）， renin， and aldosterone （ALD） levels were also measured. RESULTS ICP-MS analysis results showed statistically significant differences in the contents of macroelements Li， Na， Mg， Ca， Mn， Ga， Sr， Mo， Cd， Sn， and Sb， between the powder-directly-decocted decoction and the pieces-decocted-first decoction （ P ＜0.05） ，the elements P， Cr， Fe， Ni， Zn， Hg， Tl， and Pb were not detected in either decoction. Animal experiments showed that after two weeks of administration， compared with the model group， the facial temperature， and blood pressure decreased in all treatment groups， while the pressure pain threshold increased； plasma levels of Ang Ⅱ， renin and ALD， as well as the serum level of NO were all decreased， and thoracic aortic media thickness was significantly reduced， most of the differences in the above indicators were statistically significant （ P ＜0.05 or P ＜0.01 or P ＜0.001）. Pathological observation showed improvement in thoracic aortic pathological injury. CONCLUSIONS The powder-directly-decocted process for the Ostreae Concha- Haliotidis Concha- Margaritifera Concha drug pair significantly promotes the dissolution of key elements such as Ca， Mg， and Sr without increasing the dissolution of harmful elements. It is equivalent to the traditional pieces-decocted-first in alleviating liver-yang hyperactivity syndrome， lowering blood pressure， and protecting the vascular endothelium， and even shows better performance in some indicators.

BACKGROUND: The use of potentially inappropriate medications (PIMs) is a major concern for medication safety as it may entail more harm than potential benefits for older adults. This study aimed to explore the prescribing rate, healthcare utilization, and expenditure of older adults using PIMs in China. METHODS: A cross-sectional analysis was conducted using a national representative database of all medical insurance beneficiaries across China, extracting ambulatory visit records of adults aged 65 years and above between 2015 and 2017. Descriptive analysis was conducted to measure the rate of patients exposed to PIM, prescribing rate of each PIM, average annual outpatient visits per patient, average total medication costs for each visit, average annual cost of PIMs for each patient, and average annual medication costs for each patient. Generalized linear model with logit link function and binomial distribution was used to examine the adjusted associations between PIMs and independent variables. RESULTS: In total, 845,278 (33.2%) participants were identified to be exposed to at least one PIM. Patients aged 75-84 years (38.1%, 969,809/2,545,430) and ≥85 years (37.9%, 964,718/2,545,430) were more likely to be prescribed with PIMs. Beneficiaries of the Urban Employee Basic Medical Insurance (UEBMI) and living in eastern and southern regions were more frequently prescribed with PIMs. Compared with patients without PIM exposure (7.5 visits, drug cost of RMB 1545.0 Yuan), patients with PIM exposure showed higher adjusted average annual number of outpatient visits (10.7 visits, β = 3.228, 95% confidence interval [CI] = 3.196-3.261) and higher annual drug costs (RMB 2461.8 Yuan, Coef. = 916.864, 95% CI = RMB 906.292-927.436 Yuan). CONCLUSIONS The results showed that the use of PIM among older adults was common in China. This study suggests that the use of PIM could be considered as a clear target, pending multidimensional efforts, to promote rational prescribing for older adults.
Humans ; Aged ; Cross-Sectional Studies ; Aged, 80 and over ; Male ; Female ; China ; Inappropriate Prescribing/economics* ; Patient Acceptance of Health Care/statistics & numerical data* ; Potentially Inappropriate Medication List/statistics & numerical data* ; Health Expenditures/statistics & numerical data*

BACKGROUND: Bile acids (BAs) facilitate the progression of gastric intestinal metaplasia (GIM). Long non-coding RNAs (lncRNAs) dysregulation was observed along with the initiation of gastric cancer. However, how lncRNAs function in GIM remains unclear. This study aimed to explore the role and mechanism of lncRNA PVT1 in GIM, and provide a potential therapeutic target for GIM treatment. METHODS: We employed RNA sequencing (RNA-seq) to screen dysregulated lncRNAs in gastric epithelial cells after BA treatment. Bioinformatics analysis was conducted to reveal the regulatory mechanism. PVT1 expression was detected in 21 paired biopsies obtained under endoscopy. Overexpressed and knockdown cell models were established to explore gene functions in GIM. Molecular interactions were validated by dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (Ch-IP). The levels of relative molecular expression were detected in GIM tissues. RESULTS: We confirmed that lncRNA PVT1 was upregulated in BA-induced GIM model. PVT1 promoted the expression of intestinal markers such as CDX2 , KLF4 , and HNF4α . Bioinformatics analysis revealed that miR-34b-5p was a putative target of PVT1 . miR-34b-5p mimics increased CDX2 , KLF4 , and HNF4α levels. Restoration of miR-34b-5p decreased the pro-metaplastic effect of PVT1 . The interactions between PVT1 , miR-34b-5p, and the downstream target HNF4α were validated. Moreover, HNF4α could transcriptionally activated PVT1 , sustaining the GIM phenotype. Finally, the activation of the PVT1 /miR-34b-5p/ HNF4α loop was detected in GIM tissues. CONCLUSIONS BAs facilitate GIM partially via a PVT1/miR-34b-5p/HNF4α positive feedback loop. PVT1 may become a novel target for blocking the continuous development of GIM and preventing the initiation of gastric cancer in patients with bile reflux.
Humans ; RNA, Long Noncoding/metabolism* ; MicroRNAs/metabolism* ; Hepatocyte Nuclear Factor 4/genetics* ; Bile Acids and Salts ; Kruppel-Like Factor 4 ; Metaplasia/metabolism*

Lirispirolides A-L (1-12), twelve novel sesquiterpene-monoterpene heterodimers featuring distinctive carbon skeletons, were isolated from the branches and leaves of Chinese tulip tree [Liriodendron chinense (L. chinense)], a rare medicinal and ornamental plant endemic to China. The structural elucidation was accomplished through comprehensive spectroscopic analyses, quantum-chemical calculations, and X-ray crystallography. These heterodimers exhibit a characteristic 2-oxaspiro[4.5]decan-1-one structural motif, biosynthetically formed through intermolecular [4 + 2]-cycloaddition between a germacrane-type sesquiterpene and an ocimene-type monoterpene. The majority of the isolated compounds demonstrated significant anti-neuroinflammatory effects in lipopolysaccharide (LPS)-induced BV-2 microglial cells by reducing the production of pro-inflammatory mediators, specifically tumor necrosis factor-α (TNF-α) and nitric oxide (NO). Further investigation revealed that the lirispirolides' inhibition of NO release correlated with decreased messenger ribonucleic acid (mRNA) expression of inducible NO synthase (iNOS).
Sesquiterpenes/isolation & purification* ; Anti-Inflammatory Agents/isolation & purification* ; Animals ; Mice ; Tumor Necrosis Factor-alpha/genetics* ; Nitric Oxide/immunology* ; Microglia/immunology* ; Molecular Structure ; Liriodendron/chemistry* ; Monoterpenes/isolation & purification* ; Plants, Medicinal/chemistry* ; Cell Line ; Lipopolysaccharides ; Nitric Oxide Synthase Type II/immunology* ; Plant Extracts/pharmacology* ; China

Objective:To evaluate the potential causal relationship between tinnitus and the risk of Alzheimer's disease(AD)onset using the two-sample Mendelian randomization(MR)method and to clarify its mechanism of action,so as to provide new ideas for early warning of AD.Methods:Genome-wide association study(GWAS)database was used to search the keywords"tinnitus"and"Alzheimer"to obtain the related datasets of exposure factor tinnitus and outcome AD;the tinnitus datasets included ukb-d-4803_11,ukb-d-4803_12,ukb-d-4803_13,ukb-b-14254 and ukb-a-384;the AD datasets included ieu-b-5067,ieu-b-2,ieu-a-297,ebi-a-GCST90027158 and ebi-a-GCST002245.The single nucleotide polymorphisms(SNPs)closely and independently associated with tinnitus were screened as instrumental variables(IVs),and the SNPs associated with AD were used as outcomes.Inverse variance weighted(IVW)method was used to conduct MR analysis to evaluate its odds ratio(OR)value,95%confidence interval(CI)and P value;P<0.05 indicated significant causal relationship.Sensitivity detection used Cochran's Q test to detect the heterogeneity of IVs to evaluate its Q value,df value and P value;when IVW method P>0.05,it indicated no significant heterogeneity;MR-Egger intercept was used to detect horizontal pleiotropy;when the intercept was 0 or close to 0 and P>0.05,it indicated no significant horizontal pleiotropy;meanwhile,leave-one-out method was used for sensitivity analysis.Finally,visualization results were performed using forest plot,scatter plot,funnel plot and leave-one-out plot.Results:A total of 286 SNPs were screened as IVs.All instrumental variables satisfied F>10,suggesting no weak instrumental variable;after screening by PhenoScanner web tool,all SNPs were unrelated to confounding factors.When the tinnitus and AD datasets were ukb-d-4803 and ebi-a-GCST90027158 respectively,there was a significant positive correlation between tinnitus and the risk of AD onset(IVW:OR=1.842,95%CI:1.065-3.188,P=0.029);Cochran's Q test suggested no significant heterogeneity of IVs(Q=9.788,df=10.000,P=0.459);MR-Egger intercept indicated no horizontal pleiotropy(Egger intercept=-0.006,P=0.147);leave-one-out method showed stable results,and no SNP with significant influence on the results was detected.Conclusion:There is a positive causal relationship between tinnitus and the risk of AD onset.Neuroinflammation accompanied by persistent microglial activation to varying degrees may be the common pathogenesis of tinnitus and AD;in addition,depression may also act as an upstream factor to hyperactivate the hypothalamic-pituitary-adrenal(HPA)axis,leading to the progression of relationship between tinnitus and AD.