Helicobacter pylori CagA Promotes the Formation of Gallstones by Increasing the Permeability of Gallbladder Epithelial Cells.

BACKGROUND: The formation of gallstones is often accompanied by chronic inflammation, and the mechanisms underlying inflammation and stone formation are not fully understood. Our aim is to utilize single-cell transcriptomics, bulk transcriptomics, and microbiome data to explore key pathogenic bacteria that may contribute to chronic inflammation and gallstone formation, as well as their associated mechanisms. METHODS: scRNA-seq data from a gallstone mouse model were extracted from the Gene Expression Omnibus (GEO) database and analyzed using the FindCluster() package for cell clustering analysis. Bulk transcriptomics data from patients with gallstone were also extracted from the GEO database, and intergroup functional differences were assessed using GO and KEGG enrichment analysis. Additionally, 16S rRNA sequencing was performed on gallbladder mucosal samples from asymptomatic patients with gallstone (n = 6) and liver transplant donor gallbladder mucosal samples (n = 6) to identify key bacteria associated with stone formation and chronic inflammation. Animal models were constructed to investigate the mechanisms by which these key pathogenic bacterial genera promote gallstone formation. RESULTS: Analysis of scRNA-seq data from the gallstone mouse model (GSE179524) revealed seven distinct cell clusters, with a significant increase in neutrophil numbers in the gallstone group. Analysis of bulk transcriptomics data from patients with gallstone (GSE202479) identified chronic inflammation in the gallbladder, potentially associated with dysbiosis of the gallbladder microbiota. 16S rRNA sequencing identified Helicobacter pylori as a key bacterium associated with gallbladder chronic inflammation and stone formation. CONCLUSIONS: Dysbiosis of the gallbladder mucosal microbiota is implicated in gallstone disease and leads to chronic inflammation. This study identified H. pylori as a potential key mucosal resident bacterium contributing to gallstone formation and discovered its key pathogenic factor CagA, which causes damage to the gallbladder mucosal barrier. These findings provide important clues for the prevention and treatment of gallstones.

Bibliometric analysis of transforaminal lumbar interbody fusion: research status, trends, and future directions.

Purpose: Transforaminal lumbar interbody fusion (TLIF) is a classic surgical procedure for posterior lumbar fusion. This study aims to analyze the TLIF field by bibliometric method and comprehensively summarize the research status and trends. Methods: All TLIF-related articles were retrieved from the Web of Science. The data were analyzed using R software and SPSS to calculate corresponding indicators. Visualizations were drawn using VOSviewer and Scimago Graphica, including country, institution, journal, author, and keywords. Results: A total of 919 articles were included. The annual publication volume of TLIF-related articles presented an exponential growth. North America, Europe, and Asia were the main sources of articles, with the USA and China being the main contributors and the USA being the global research center for TLIF. The level of the national economy was an important factor affecting TLIF-related research. The highest number of contributions in this field was made by Kern Singh among authors and by Rush University among institutions. The European Spine Journal was the most influential journal. The research focus has gradually shifted from perfecting the TLIF technique toward emphasizing the patient level. The improvement of minimally invasive techniques and how to improve clinical outcomes as well as accelerate postoperative rehabilitation of patients may be the hot spot of future research. Conclusions: With the advancement of medical technology and the popularization of minimally invasive concepts in recent years, TLIF and its derivative technologies have attracted increasing attention. Patient-centered minimally invasive surgery is a hot research topic in the field of TLIF currently and will continue to be so into the future.

Causality between urate levels with sarcopenia-related traits: a bi-directional Mendelian randomization study.

Background: Observational studies have suggested associations between serum urate levels and sarcopenia, but the causality underlying this correlation remains uncertain. The principal objective of this study is to investigate a causal relationship of serum urate levels with sarcopenia-related traits (hand grip strength, lean mass, walking pace) using bidirectional two-sample Mendelian randomization (MR) approach. The utilization of MR methodology serves to minimize bias caused by reverse causality and confounding factors from observational studies. Methods: The summary statistics of serum urate levels were derived from a cohort consisting of 288,659 individuals participating in CKDGen study. The parameters of right-hand grip strength (N=461,089), left-hand grip strength (N=461,026), appendicular lean mass (ALM) (N=450,243), whole-body lean mass(N=454,850),right-leg fat-free mass(FFM;N=454,835),left-leg FFM(N=454,805), right-arm FFM(N=454,753),left-arm FFM(N=454,672) and walking pace (N=459,915)were sourced from the UK Biobank. MR analysis was conducted utilizing inverse variance weighted (IVW), weighted median, and MR-Egger to evaluate causality. Sensitivity analysis was performed using Cochran's Q test, MR-Egger intercept test, leave-one-out analysis and the funnel plot. Results: IVW estimates demonstrated that serum urate levels exhibited no causal association with sarcopenia-related traits. In the inverse MR investigation, we had exclusively discerned an inverse correlation between walking pace and serum urate levels. No compelling evidence had surfaced to substantiate any association of other sarcopenia-related traits with serum urate. Supplemental MR methods consistently validated the findings obtained from the primary analysis. Sensitivity analysis demonstrated the robustness of findings. Conclusion: Our MR study revealed the absence of the bidirectional causal relationship between serum urate levels and sarcopenia. It is imperative to acknowledge that advanced age and an individual's health status are pivotal determinants influencing urate level and the initiation and advancement of sarcopenia. However, it is worth underscoring that these aspects remain unexamined within the purview of this study. Thus, future investigations should delve deeper into these intricate facets.

Association of visceral and subcutaneous fat with bone mineral density in US adults: a cross-sectional study.

The relationship between the accumulation of fat in visceral or subcutaneous tissue and bone mineral density (BMD) remains unclear. Our primary objective in this study was to illuminate this relationship by conducting an investigation on a vast scale, encompassing a nationally representative population in the United States. A weighted multiple linear regression model was established to evaluate the relationship between visceral fat, subcutaneous fat, and BMD. Additionally, the exploration of the potential nonlinear relationship was conducted employing the methodology of smooth curve fitting. In order to determine potential inflection points, a two-stage linear regression model was utilized. A total of 10,455 participants between the ages of 20 and 59 were included in this study. Various weighted multiple linear regression models revealed a negative correlation between lumbar BMD and visceral mass index (VMI) and subcutaneous mass index (SMI). However, the association between VMI and lumbar BMD displayed a U-shaped pattern upon employing the smooth curve fitting, and the inflection point of 0.304 kg/m2was determined using a two-stage linear regression model. Our findings indicated a negative association between subcutaneous fat and BMD. A U-shaped relationship was observed between visceral fat and BMD.

Association between the use of lipid-lowering drugs and the risk of inflammatory bowel disease.

BACKGROUND: Observational studies have suggested an association between lipid-lowering drugs and inflammatory bowel disease (IBD) risk. This study aimed to assess the causal influence of lipid-lowering agents on IBD risk using Mendelian randomization analysis. METHOD: In a population of 173,082 individuals of European ancestry, 55 single-nucleotide polymorphisms were identified as instrumental variables for 6 lipid-lowering drug targets (HMGCR, NPC1LC, PCSK9, LDLR, CETP and APOB). Summary statistics for the genome-wide association study of IBD, ulcerative colitis (UC) and Crohn's disease (CD) were obtained from the FinnGen consortium, Program in Complex Trait Genomics and UK Biobank. Inverse-variance weighted was employed as the primary MR method, and odds ratios (ORs) with 95% confidence intervals were reported as the results. Sensitivity analyses using conventional MR methods were conducted to assess result robustness. RESULTS: Gene-proxied inhibition of Niemann-Pick C1-like 1 (NPC1L1) was associated with an increased IBD risk (OR [95% CI]: 2.31 [1.38, 3.85]; p = .001), particularly in UC (OR [95% CI]: 2.40 [1.21, 4.74], p = .012), but not in CD. This finding was replicated in the validation cohort. Additionally, gene-proxied inhibition of low-density lipoprotein receptor was associated with reduced IBD (OR [95% CI]: .72 [.60, .87], p < .001) and UC risk (OR [95% CI]: .74 [.59, .92], p = .006), although this result was not replicated in the validation cohort. Other drug targets did not show significant associations with IBD, UC or CD risk. CONCLUSION: Inhibition of the lipid-lowering drug-target NPC1L1 leads to an increased IBD risk, mainly in the UC population.

Surgical site infection in spinal surgery: a bibliometric analysis.

BACKGROUND: Surgical site infection (SSI) is a common complication in spinal surgery that can significantly affect the patient's prognosis. Despite advances in surgical techniques and infection control measures, SSI remains a considerable concern for healthcare providers and patients alike. In recent years, there has been a steady increase in studies related to SSI in spine surgery, leading to the publication of numerous informative articles. However, the current state and trends of research in the field of spinal SSI remain unclear. This study aims to conduct a bibliometric analysis of SSI-related articles in spine surgery to identify research status and trends. Meanwhile, we identify the top 100 most cited articles for further analysis. METHODS: We searched for all articles related to spinal SSI in the Web of Science Core Collection, recording the publication year, country, journal, institution, keywords, and citation frequency for further analysis. In addition, we identified and analyzed the top 100 most cited articles. RESULTS: A total of 307 articles related to spinal SSI were identified. All of these articles were published between 2008 and 2022, with the number of publications showing an increasing trend over the years. The related articles originated from 37 countries, with the USA contributing the most (n = 138). The institution with the highest number of publications and citations was Johns Hopkins University (14 articles; 835 citations). Among the journals, Spine had the highest number of articles (n = 47). The prevention of spinal SSI has been a research hotspot in recent years. Among the top 100 most cited articles, the most common research theme was the risk factors associated with spinal SSI. CONCLUSIONS: In recent years, research related to spinal SSI has attracted the attention of numerous clinicians and scholars. As the first bibliometric analysis of spinal SSI, our study aims to provide pragmatic guidance for clinicians to learn the research status and trends in this field and improve their vigilance toward SSI.

Combination Analysis of Ferroptosis and Immune Status Predicts Patients Survival in Breast Invasive Ductal Carcinoma.

Ferroptosis is a new form of iron-dependent cell death and plays an important role during the occurrence and development of various tumors. Increasingly, evidence shows a convincing interaction between ferroptosis and tumor immunity, which affects cancer patients' prognoses. These two processes cooperatively regulate different developmental stages of tumors and could be considered important tumor therapeutic targets. However, reliable prognostic markers screened based on the combination of ferroptosis and tumor immune status have not been well characterized. Here, we chose the ssGSEA and ESTIMATE algorithms to evaluate the ferroptosis and immune status of a TCGA breast invasive ductal carcinoma (IDC) cohort, which revealed their correlation characteristics as well as patients' prognoses. The WGCNA algorithm was used to identify genes related to both ferroptosis and immunity. Univariate COX, LASSO regression, and multivariate Cox regression models were used to screen prognostic-related genes and construct prognostic risk models. Based on the ferroptosis and immune scores, the cohort was divided into three groups: a high-ferroptosis/low-immune group, a low-ferroptosis/high-immune group, and a mixed group. These three groups exhibited distinctive survival characteristics, as well as unique clinical phenotypes, immune characteristics, and activated signaling pathways. Among them, low-ferroptosis and high-immune statuses were favorable factors for the survival rates of patients. A total of 34 differentially expressed genes related to ferroptosis-immunity were identified among the three groups. After univariate, Lasso regression, and multivariate stepwise screening, two key prognostic genes (GNAI2, PSME1) were identified. Meanwhile, a risk prognosis model was constructed, which can predict the overall survival rate in the validation set. Lastly, we verified the importance of model genes in three independent GEO cohorts. In short, we constructed a prognostic model that assists in patient risk stratification based on ferroptosis-immune-related genes in IDC. This model helps assess patients' prognoses and guide individualized treatment, which also further eelucidatesthe molecular mechanisms of IDC.

Impacts of core training on athletes' performance in long-distance running / Impactos do treino do core no desempenho de atletas na corrida de longa distância / Efectos del entrenamiento del core en el rendimiento de atletas en carreras de larga distancia

ABSTRACT Introduction: Long-distance runners cannot achieve training effects by relying only on speed endurance training. Core training exercises significantly improve stability, allowing learning of techniques with superior movement efficiency. Several factors that affect the development of athletes' special skills are considered, highlighting their main strengths and working on intermediate characteristics. Objective: Verify the impacts of core training on athletes' performance in long-distance running. Methods: This paper had 18 middle-distance runners as volunteers for the research. The athletes' physical conditioning and athletic performance before and after training were examined. Statistical analyses on the indicators of the experimental and control groups were analyzed based on the T-test. Results: There were significant differences in endurance, core muscle elasticity, core strength, explosive power, and stability (P<0.05); the results showed that the differences in core stability between the left and right legs of the experimental group were significant (P<0.05). Conclusion: Core strength exercise is more conducive to exercising a myocardial group in medium and long-distance exercise than conventional strength exercise. Core strength exercises can compensate for a lack of regular strength. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: Corredores de longa distância não podem alcançar efeitos de treinamento apenas contando com os treinos de resistência de velocidade. Os exercícios de treino do core melhoram significativamente a estabilidade que permite um aprendizado da técnica com eficiência superior do movimento. Para isso, são considerados vários fatores que afetam o desenvolvimento das habilidades especiais dos atletas, destacando seus principais pontos fortes e trabalhando nas características intermediárias. Objetivo: Verificar os impactos do treino do core no desempenho de atletas na corrida de longa distância. Métodos: Este artigo teve a participação de 18 corredores de média distância como voluntários para a pesquisa. Foi examinado o condicionamento físico dos atletas e o desempenho atlético antes e depois dos treinos. As análises estatísticas nos indicadores dos grupos experimentais e de controle foram analisadas com base no teste-T. Resultados: Houve diferenças significativas na resistência, elasticidade muscular essencial, força do core, potência explosiva e estabilidade (P<0,05); os resultados mostraram que as diferenças na estabilidade do núcleo entre as pernas esquerda e direita do grupo experimental foram significativas (P<0,05). Conclusão: O exercício de força do core é mais propício ao exercício de um grupo miocárdio em exercícios de média e longa distância do que o exercício de força convencional. Exercícios de força do core podem compensar a falta de força regular. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: Los corredores de larga distancia no pueden conseguir los efectos del entrenamiento basándose únicamente en el entrenamiento de la velocidad y la resistencia. Los ejercicios de entrenamiento del core mejoran significativamente la estabilidad que permite un aprendizaje de la técnica con una eficiencia de movimiento superior. Para ello, se tienen en cuenta varios factores que afectan al desarrollo de las capacidades especiales de los deportistas, destacando sus principales puntos fuertes y trabajando sobre las características intermedias. Objetivo: Verificar los impactos del entrenamiento del core en el rendimiento de los atletas en carreras de larga distancia. Métodos: Este artículo contó con la participación de 18 corredores de media distancia como voluntarios para la investigación. Se examinó la condición física y el rendimiento deportivo de los atletas antes y después del entrenamiento. Los análisis estadísticos en los indicadores de los grupos experimental y de control se analizaron a partir de la prueba T. Resultados: Hubo diferencias significativas en la resistencia, la elasticidad de los músculos centrales, la fuerza central, la potencia explosiva y la estabilidad (P<0,05); los resultados mostraron que las diferencias en la estabilidad central entre las piernas izquierda y derecha del grupo experimental eran significativas (P<0,05). Conclusión: El ejercicio de fuerza del núcleo favorece más el ejercicio del grupo miocárdico en el ejercicio de media y larga distancia que el ejercicio de fuerza convencional. Los ejercicios de fuerza del core pueden compensar la falta de fuerza habitual. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

ScRNA-seq and bulk RNA-seq reveal the characteristics of ferroptosis and establish a risk signature in cholangiocarcinoma.

Ferroptosis is a recently discovered mode of cell death that inhibits tumor growth. Single-cell RNA sequencing (scRNA-seq) is a powerful tool for analyzing tumor heterogeneity and the immune microenvironment at the single-cell level. We used CIBERSORT to identify cellular immune scores and found that monocytes had significantly infiltrated and were correlated with prognosis in cholangiocarcinoma. scRNA-seq data were extracted from the Gene Expression Omnibus database, and the FindCluster() package was used for cell cluster analysis, which obtained 21 cell clusters, and there was increased TNFSF13B-TFRC intercellular communication between monocytes and cholangiocytes. A weighted correlation network analysis was performed with the WGCNA package to obtain monocyte-related gene modules. Univariate and multivariate Cox analyses were then performed to further establish the signature, and the reliability of the signature was assessed by receiver operating characteristic curve and decision curve analysis. A nomogram signature based on the Kaplan-Meier survival analysis was established. We found that the communication between monocytes and malignant cells in cholangiocarcinoma may be a regulatory factor of ferroptosis in cancer cells. The prognostic stratification system of the three-gene signature related to monocytes and ferroptosis can accurately assess the prognostic risk for cholangiocarcinoma.

The Rabep1-Mediated Endocytosis and Activation of Trypsinogen to Promote Pancreatic Stellate Cell Activation.

BACKGROUND: The pathogenesis of chronic pancreatitis is still unclear. Trypsinogen activation is an active factor in acute pancreatitis that has not been studied in the occurrence of chronic pancreatitis. METHODS: Immunofluorescence was used to detect the location and expression of trypsinogen in chronic pancreatitis and normal tissues. Microarray and single-cell RNA-seq (scRNA-seq) were used to screen core genes and pathways in pancreatic stellate cells (PSCs). Western blotting and immunofluorescence were used to verify trypsinogen expression in PSCs after silencing Rabep1. Immunofluorescence and flow cytometry were used to validate trypsinogen activation and PSC activation after intervening in the endocytosis pathway. RESULTS: Endocytosed trypsinogen was found in PSCs in CP clinical samples. Bioinformatic analysis showed that Rabep1 is a core gene that regulates trypsinogen endocytosis through the endocytosis pathway, verified by Western blot and immunofluorescence. Immunofluorescence and flow cytometry analyses confirmed the activation of trypsinogen and PSCs through the endocytosis pathway in PSCs. CONCLUSION: This study discovered a new mechanism by which trypsinogen affects the activation of PSCs and the occurrence and development of CP. Through communication between pancreatic acinar cells and PSCs, trypsinogen can be endocytosed by PSCs and activated by the Rabep1 gene.

A novel HCC prognosis predictor PDSS1 affects the cell cycle through the STAT3 signaling pathway in HCC.

Decaprenyl diphosphate synthase subunit 1 (PDSS1) is closely related to a variety of human diseases, but its expression pattern and biological function in HCC have not been studied to date. Methods: The expression level of PDSS1 was analyzed using the TCGA and GEO databases. The relationships between PDSS1 and patient clinicopathological characteristics were verified based on TCGA clinical data. Additionally, the co-expressed genes of PDSS1were investigated and Gene Set Enrichment Analysis (GSEA) was conducted using LinkedOmics. Next, the association between PDSS1 and immune infiltration was determined using version 1.34.0 of the GSVA package. EdU assay, colony-formation assay, transwell assay, wound-healing assay, and flow cytometry analysis were used to assess the effect of PDSS1 on the cell phenotype. Results: PDSS1 was upregulated in HCC compared with adjacent tissues. High PDSS1 in HCC was associated with poor overall survival, disease-specific survival, and progress-free interval. Results suggested that PDSS1 may activate multiple oncogenic pathways in HCC, especially those involved in the cell cycle. The expression of PDSS1 was significantly related to Th2 cells, TFH, T helper cells, NK CD56bright cells, cytotoxic cells, DC, CD8 T cells, and neutrophils. PDSS1 knockdown inhibited cell proliferation, cell cycle, migration and invasion. Furthermore, PDSS1 acted as an oncogene through the STAT3 signaling pathway. Conclusion: Our study reveals that a high level of PDSS1 is significantly correlated with poor patient prognosis and immune cell infiltration in HCC. PDSS1 may be a novel biomarker and potential therapeutic target for HCC.

USH2A Mutation is Associated With Tumor Mutation Burden and Antitumor Immunity in Patients With Colon Adenocarcinoma.

Colon adenocarcinoma (COAD) is one of the diseases with the highest morbidity and mortality in the world. At present, immunotherapy has become a valuable method for the treatment of COAD. Tumor mutational burden (TMB) is considered to be the most common biomarker for predicting immunotherapy. According to reports, the mutation rate of COAD ranks third. However, whether these gene mutations are related to TMB and immune response is still unknown. Here, COAD somatic mutation data were downloaded from The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) databases. Bioinformatics methods were used to study the relationships among gene mutations, COAD survival prognosis, and tumor immune response. A total of 22 of the top 40 mutations in TCGA and ICGC databases were the same. Among them, the USH2A mutation was associated with high TMB and poor clinical prognosis. According to Gene Set Enrichment Analysis (GSEA) and the CIBERSORT algorithm, we determined that the USH2A mutation upregulates signaling pathways involved in the immune system and the antitumor immune response. In cases with a USH2A mutation, the immune score and MSI score of TCGA samples increased, the expression of immune checkpoint genes decreased significantly, and the TIDE score decreased significantly. Dependent on the presence or absence of a USH2A mutation, TCGA COAD samples were analyzed for differentially expressed genes, 522 of which were identified. Using a univariate Cox analysis and LASSO COX analysis of these differential genes, a prediction model was established, which established significant differences in the infiltration of immune cells, immune checkpoint gene expression, immune score, MSI score, TMB, and TIDE in patients in high- and low-risk groups. In conclusion, mutation of USH2A is frequent in COAD and is related to an increase in TMB and the antitumor immunity. The differential genes screened by USH2A mutation allowed the construction of a risk model for predicting the survival and prognosis of cancer patients, in addition to providing new ideas for COAD immunotherapy.

The nomogram based on the 6-lncRNA model can promote the prognosis prediction of patients with breast invasive carcinoma.

Long non-coding RNA (lncRNA) is a prognostic biomarker for many types of cancer. Here, we aimed to study the prognostic value of lncRNA in Breast Invasive Carcinoma (BRCA). We downloaded expression profiles from The Cancer Genome Atlas (TCGA) datasets. Subsequently, we screened the differentially expressed genes between normal tissues and tumor tissues. Univariate Cox, LASSO regression, and multivariate Cox regression analysis were used to construct a lncRNA prognostic model. Finally, a nomogram based on the lncRNAs model was developed, and weighted gene co-expression network analysis (WGCNA) was used to predict mRNAs related to the model, and to perform function and pathway enrichment. We constructed a 6-lncRNA prognostic model. Univariate and multivariate Cox regression analysis showed that the 6-lncRNA model could be used as an independent prognostic factor for BRCA patients. We developed a nomogram based on the lncRNAs model and age, and showed good performance in predicting the survival rates of BRCA patients. Also, functional pathway enrichment analysis showed that genes related to the model were enriched in cell cycle-related pathways. Tumor immune infiltration analysis showed that the types of immune cells and their expression levels in the high-risk group were significantly different from those in the low-risk group. In general, the 6-lncRNA prognostic model and nomogram could be used as a practical and reliable prognostic tool for invasive breast cancer.

Pan-Cancer Analysis of Prognostic and Immune Infiltrates for CXCs.

BACKGROUND: CXCs are important genes that regulate inflammation and tumor metastasis. However, the expression level, prognosis value, and immune infiltration of CXCs in cancers are not clear. METHODS: Multiple online datasets were used to analyze the expression, prognosis, and immune regulation of CXCs in this study. Network analysis of the Amadis database and GEO dataset was used to analyze the regulation of intestinal flora on the expression of CXCs. A mouse model was used to verify the fact that intestinal bacterial dysregulation can affect the expression of CXCs. RESULTS: In the three cancers, multiple datasets verified the fact that the mRNA expression of this family was significantly different; the mRNA levels of CXCL3, 8, 9, 10, 14, and 17 were significantly correlated with the prognosis of three cancers. CXCs were correlated with six types of immuno-infiltrating cells in three cancers. Immunohistochemistry of clinical samples confirmed that the expression of CXCL8 and 10 was higher in three cancer tissues. Animal experiments have shown that intestinal flora dysregulation can affect CXCL8 and 10 expressions. CONCLUSION: Our results further elucidate the function of CXCs in cancers and provide new insights into the prognosis and immune infiltration of breast, colon, and pancreatic cancers, and they suggest that intestinal flora may influence disease progression through CXCs.

Intestinal flora imbalance affects bile acid metabolism and is associated with gallstone formation.

BACKGROUND: The gut microbiota participates in the metabolism of substances and energy, promotes the development and maturation of the immune system, forms the mucosal barrier, and protects the host from pathogen attacks. Although the pathogenesis of cholesterol gallstones is still not clear, studies have suggested that gut microbiota dysbiosis plays an important role in their formation. METHODS: Microbial DNA from faeces of normal control patients and those of patients with calculi was subjected to 16S rRNA gene sequencing to detect gene expression changes in intestinal microbes. ELISA kits were used to measure free bile acids, secondary bile acids and coprostanol according to the manufacturer's instructions. The relationship between flora and their metabolites was then analysed. RESULTS: In the gallstone group, the diversity of intestinal bacteria and the abundances of certain phylogroups were significantly decreased (p < 0.05), especially Firmicutes (p < 0.05), the largest phylum represented by the gut microbiota. This study found an increase in free bile acids (p < 0.001) and secondary bile acids (p < 0.01) in the enterohepatic circulation. Bile salt hydrolase activity was not related to the abundances of BSH-active bacteria. 7a-dehydroxylating gut bacteria were significantly increased (p < 0.01), whereas cholesterol-lowering bacteria were significantly reduced (p < 0.05). The Ruminococcus gnavus group could be used as a biomarker to distinguish the gallstone group from the control group. CONCLUSION: We conclude that intestinal flora imbalance affects bile acid and cholesterol metabolism and is associated with gallstone formation.

The Regulatory Effect of the Kinase Inhibitor PD98059 on Autophagic Flux During Trypsinogen Activation in Pancreatic Acinar Cells.

OBJECTIVES: To study the role of kinase inhibitor PD98059 on autophagy flow in the process of trypsinogen activation in pancreatic acinar cell and its related mechanism. METHODS: In the present study, bioinformatics analysis was used to predict kinases and their most relevant inhibitor (PD98059) which participates in autophagy of acute pancreatitis (AP). The rat pancreatic acini AR42J cells were divided into 4 groups: control group, sodium taurocholate hydrate (TLC) group, PD98059 group, and TLC + PD group. Twenty-seven Sprague-Dawley rats were divided into 3 groups (n = 9), including control group, severe AP (SAP) group, and SAP + PD group. We detected trypsinogen activation, autophagic activation, lysosome pH, and cathepsin-L activity in vivo and in vitro. RESULTS: Results revealed trypsinogen activation was significantly inhibited in mitogen-activated protein kinase 1, JAK2, LYN, and their common inhibitor was PD98059. The trypsinogen activation, Beclin1, and light chain 3 II expressions were reduced, whereas the expressions of lysosomal-associated membrane protein 2, cathepsin L1, and cathepsin-L activity is upregulated after the PD98059 pretreatment, both in vivo and in vitro. CONCLUSIONS: Lysosomal dysfunction blocked autophagy flux, accompanied by increasing pancreatic acinar cell autophagy in the process of trypsinogen activation. PD98059 inhibited AP occurrence and pancreatic injury via improving the blocked autophagic pathway and reducing trypsinogen activation.