ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley （SD） rats were randomly divided into a normal group， model group （M）， Sini San group （MS， 2.5 g·kg^-1）， and fluoxetine group （MF， 2 mg·kg^-1）. Except for the normal group， rats in the other three groups were subjected to chronic unpredictable mild stress （CUMS）. After 8 weeks， the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum corticosterone （CORT）， adrenocorticotropic hormone （ACTH）， corticotropin-releasing factor （CRF）， lipopolysaccharide （LPS）， Zonulin， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， γ-aminobutyric acid （GABA） levels in the hippocampus and prefrontal cortex， and brain-derived neurotrophic factor （BDNF） levels in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy， and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group， the sucrose preference of rats in the model group was significantly reduced （P0.01）， whereas it was significantly increased in the Sini San group compared with the model group （P0.05）. Compared with the normal group， hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased （P0.05）， and compared with the model group， both were significantly increased in the Sini San group （P0.05， P0.01）. Compared with the normal group， serum LPS and Zonulin levels in the model group were significantly increased （P0.05， P0.01）， and compared with the model group， Zonulin levels in the Sini San group were significantly decreased （P0.05）. No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group， widened and blurred tight junctions， sparse and shortened microvilli， and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group， which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators， liver function， and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.

ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley （SD） rats were randomly divided into a normal group， model group （M）， Sini San group （MS， 2.5 g·kg^-1）， and fluoxetine group （MF， 2 mg·kg^-1）. Except for the normal group， rats in the other three groups were subjected to chronic unpredictable mild stress （CUMS）. After 8 weeks， the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum corticosterone （CORT）， adrenocorticotropic hormone （ACTH）， corticotropin-releasing factor （CRF）， lipopolysaccharide （LPS）， Zonulin， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， γ-aminobutyric acid （GABA） levels in the hippocampus and prefrontal cortex， and brain-derived neurotrophic factor （BDNF） levels in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy， and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group， the sucrose preference of rats in the model group was significantly reduced （P<0.01）， whereas it was significantly increased in the Sini San group compared with the model group （P<0.05）. Compared with the normal group， hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased （P<0.05）， and compared with the model group， both were significantly increased in the Sini San group （P<0.05， P<0.01）. Compared with the normal group， serum LPS and Zonulin levels in the model group were significantly increased （P<0.05， P<0.01）， and compared with the model group， Zonulin levels in the Sini San group were significantly decreased （P<0.05）. No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group， widened and blurred tight junctions， sparse and shortened microvilli， and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group， which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators， liver function， and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.

BACKGROUND: Double-stranded RNA-specific adenosine deaminase 1 (ADAR1) binds to double-stranded RNA and catalyzes the deamination of adenosine (A) to inosine (I). The functional mechanism of ADAR1 in non-small cell lung cancer (NSCLC) remains incompletely understood. This study aimed to investigate the prognostic significance of ADAR1 in NSCLC and to elucidate its potential role in regulating tumor cell proliferation and migration. METHODS: Data from The Cancer Genome Atlas (TCGA) and cBioPortal were analyzed to assess the correlation between high ADAR1 expression and clinicopathological features as well as prognosis in lung cancer. We performed Western blot (WB), cell proliferation assays, Transwell invasion/migration assays, and nude mouse xenograft modeling to examine the phenotypic changes and molecular mechanisms induced by ADAR1 knockdown. Furthermore, the ADAR1 p150 overexpression model was utilized to validate the proposed mechanism. RESULTS: ADAR1 expression was significantly elevated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tissues compared with adjacent non-tumor tissues (LUAD: P=3.70×10-15, LUSC: P=0.016). High ADAR1 expression was associated with poor prognosis (LUAD: P=2.03×10-2, LUSC: P=2.81×10-2) and distant metastasis (P=0.003). Gene Set Enrichment Analysis (GSEA) indicated that elevated ADAR1 was associated with mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway activation, matrix metalloproteinase-9 (MMP-9) expression, and cell adhesion. ADAR1 and MMP-9 levels showed a strongly positive correlation (P=6.45×10-34) in 10 lung cancer cell lines, highest in H1581. Knockdown of ADAR1 in H1581 cells induced a rounded cellular morphology with reduced pseudopodia. Concomitantly, it suppressed cell proliferation, invasion, migration, and in vivo tumorigenesis. It also suppressed ERK phosphorylation and downregulated cellular Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (c-FOS), MMP-9, N-cadherin, and Vimentin. Conversely, ADAR1 p150 overexpression in PC9 cells enhanced ERK phosphorylation and increased c-FOS and MMP-9 expression. CONCLUSIONS High ADAR1 expression is closely associated with poor prognosis and distant metastasis in NSCLC patients. Mechanistically, ADAR1 may promote proliferation, invasion, migration, and tumorigenesis in lung cancer cells via the ERK/c-FOS/MMP-9 axis.
Humans ; Lung Neoplasms/physiopathology* ; Adenosine Deaminase/genetics* ; Matrix Metalloproteinase 9/genetics* ; Cell Proliferation ; Carcinoma, Non-Small-Cell Lung/physiopathology* ; Cell Movement ; Animals ; Mice ; RNA-Binding Proteins/genetics* ; Female ; Male ; Cell Line, Tumor ; Proto-Oncogene Proteins c-fos/genetics* ; Middle Aged ; MAP Kinase Signaling System ; Gene Expression Regulation, Neoplastic ; Mice, Nude ; Extracellular Signal-Regulated MAP Kinases/genetics*

Objective:To evaluate the characteristics of neutrophil extracellular traps (NET) in children with inflammatory bowel disease (IBD) and its role in diagnosis and disease activity monitoring.Methods:A total of 66 IBD children admitted to Beijing Children′s Hospital from December 2017 to August 2024 were enrolled in this cross-sectional study, another 20 age-matched children who underwent gastrointestinal endoscopy during the same period in the same hospital and showed no abnormalities were selected as the controls. Clinical data of IBD and control group were collected. Children with IBD were divided into active group and remission group according to clinical score and endoscopic score. The peripheral blood of IBD and control group were collected, and the levels of NET markers, including neutrophil elastase (NE) and myeloperoxidase (MPO)-DNA were detected by enzyme-linked immunosorbent assay. The levels of NET markers in control group and different IBD groups were compared. Independent sample t-test or Mann-Whitney U test was used for group comparisons. Kruskal-Wallis H test was used for multiple group comparisons. Spearman correlation analysis was used to analyze the correlation between NET markers and IBD activity. The efficacy of laboratory indicators in diagnosing IBD and control group was evaluated using receiver operating characteristic (ROC) curve. Results:There were 66 children with IBD, including 36 in Crohn′s disease group with the age of (11.0±3.7) years, and 30 in ulcerative colitis (UC) group with the age of (8.3±5.0) years. The control group consisted of 20 children with the age of (10.1±3.5) years. Compared with control group, the levels of NE (958 (771, 1 328) vs. 303 (196, 501) μg/L) and MPO-DNA (0.11 (0.09, 0.18) vs. 0.09 (0.06, 0.12)) in peripheral blood of IBD group were significantly higher (both P<0.05). However, there was no significant difference in the levels of NE (1 008 (863, 1 301) vs. 807 (567, 1 535) μg/L) and MPO-DNA (0.11 (0.09, 0.21) vs. 0.12 (0.09, 0.14)) between Crohn′s disease and UC groups (both P>0.05). The NE levels in the endoscopic active group and remission group of Crohn's disease were higher than those in the control group (both P<0.05). The MPO-DNA level in the endoscopic active group of Crohn's disease was higher than that in the control group ( P<0.05), while the MPO-DNA level in the endoscopic remission group of Crohn's disease was lower than that in the control group ( P>0.05). The NE levels in the endoscopic activity group and remission group of UC were higher than those in control group (both P<0.05). NET markers were not correlated with the clinical activity and endoscopic activity of IBD (all P>0.05). ROC curve analysis showed that the area under the curve of NE combined with MPO-DNA for distinguishing IBD from controls was 0.95, with a sensitivity was 90.0% and a specificity was 89.4%. Conclusion:The combination of NE and MPO-DNA demonstrated high sensitivity and specificity for distinguishing pediatric IBD patients from healthy children, suggesting its potential as a diagnostic biomarker panel of IBD.

BACKGROUND:Mitophagy is closely associated with the development of idiopathic pulmonary fibrosis,but its mechanism remains unclear.OBJECTIVE:To investigate the biological mechanism of mitophagy in idiopathic pulmonary fibrosis and provide ideas for the risk prediction of idiopathic pulmonary fibrosis and subtype differentiation.METHODS:The mitophagy-related genes in idiopathic pulmonary fibrosis were obtained through GEO and Reactome Pathway databases.The mitophagy-related characteristic genes in idiopathic pulmonary fibrosis were screened based on intergroup differences and random forest model.GO functional enrichment analysis and KEGG,Reactome with WIKI pathway enrichment analyses were performed by g:Profiler database.Mitophagy subtypes in idiopathic pulmonary fibrosis were distinguished by consensus clustering method and immune infiltration analysis was performed.The mitophagy-related key gene was screened.Finally,the predictive value of mitophagy-related key gene for the risk of idiopathic pulmonary fibrosis was quantified by alignment diagram and the correlation between mitophagy-related key gene and clinical characteristics of idiopathic pulmonary fibrosis was explored.RESULTS AND CONCLUSION:(1)A total of 13 genes related to mitophagy in idiopathic pulmonary fibrosis were identified and 5 characteristic genes were screened,containing PINK1,RPS27A,SRC,HIF1A,and CDH6.(2)GO analysis was mainly involved in ubiquitin protein ligase binding,and cellular response to hypoxia.Pathway enrichment analysis was mainly involved in PINK1-PRKN mediated mitophagy,NOTCH signaling pathway,signaling by EGFR and angiogenesis.(3)HIF1A had significant expression differences between subtypes,which might serve as a key gene for the differentiation of mitophagy subtypes of idiopathic pulmonary fibrosis.(4)Immune infiltration analysis suggested that myeloid-derived suppressor cell,neutrophil and type 1 T helper cell might have infiltration differences between subtypes,while HIF1A was positively correlated with multiple immune cells.(5)Alignment diagram suggested that the risk of idiopathic pulmonary fibrosis might be predicted by the expression level of HIF1A.(6)Clinical characteristics analysis indicated patients with high expression of HIF1A might have poorer lung function and more severe fibrosis.It is concluded that PINK1,RPS27A,SRC,HIF1A,and CDH6 may influence the development of idiopathic pulmonary fibrosis through mitophagy,in which HIF1A may serve as a key gene for risk prediction with clinical subtype differentiation and HIF1A is strongly associated with the lung function of patients.

Objective:To investigate variation of peripheral blood monocyte subtypes in patients with rheumatoid arthritis(RA),to study variation of surface antigen expression intensity and plasma related cytokine content of monocytes of each subtype,and to discuss their clinical application value.Methods:Percentage of monocyte subtypes and mean fluorescence intensity(MFI)of monocyte surface markers(HLA-DR,CD64,CD11b)in peripheral blood of 38 patients with RA were detected by multi-parameter flow cytometry,and differences were statistically analyzed compared with 40 healthy controls(HV group).Plasma levels of five cyto-kines(IL-2,IL-6,IL-10,TNF-α and IFN-γ)were detected by flow fluorescence microsphere technology.Linear correlation analysis was performed to determine correlation between percentage of monocyte subtypes and MFI of their surface markers and expression of related cytokines in RA patients.Disease activity 28(DAS28)score was performed in RA patients to study its association with percent-age of monocyte subtypes and plasma cytokines.Results:Compared with control group,percentage of classical monocytes in RA pa-tients decreased significantly(P<0.05),while percentage of intermediate monocytes and non-classical monocytes increased signifi-cantly(P<0.01).HLA-DR expression(MFI)on monocyte subsets in RA group was significantly higher than HV group(P<0.05 or P<0.01).CD64 expression(MFI)on intermediate monocytes and non-classical monocytes in RA group was significantly higher than HV group(P<0.01 or P<0.001).CD11b expression(MFI)on intermediate monocytes in RA group was significantly higher than HV group(P<0.001).In RA group,percentage of intermediate monocytes was positively correlated with contents of IL-6,TNF-α and IFN-γ(P<0.01).HLA-DR expression(MFI)on intermediate monocytes was positively correlated with TNF-α content(P<0.05);CD64 expres-sion(MFI)on non-classical monocytes was positively correlated with contents of IL-2 and IL-10(P<0.05).In RA group,percentage of intermediate monocytes was positively correlated with DAS28(P<0.001);plasma IL-6 and TNF-α levels were positively correlated with DAS28(P<0.01).Conclusion:Characteristic expression of intermediate monocytes and their surface antigens are closely related to occurrence of RA,and related inflammation is triggered by secretion of related cytokines.Monocyte subtype detection can be a new experimental diagnostic index for RA.Dynamic monitoring of peripheral blood monocyte subtypes and plasma IL-6 and TNF-α in RA patients during treatment is helpful to guide clinical treatment of RA.

Objective To survey the awareness and intended adoption of patient-based real-time quality control(PBRTQC)among medical laboratories,thereby providing a basis for subsequent research and promotion efforts.Methods An electronic questionnaire was distributed via the Questionnaire Star system to collect participants' general information,their knowledge,attitudes,and practices regarding PBRTQC,as well as their implementation willingness.Results A total of 161 valid questionnaires were collected.The sur-vey revealed that fewer than 50％of laboratory staff had basic knowledge of PBRTQC,and their confidence in establishing and applying PBRTQC in their own laboratories was low.However,they acknowledged its potential to enhance quality management.Key concerns and support needs for PBRTQC promotion include professional implementation guidelines and policy support,reliable third-party evalu-ations,software and hardware support,and user training.Conclusion The laboratory staff accepted the value of PBRTQC and gener-ally held a positive attitude toward its application.Future efforts should focus on developing standardized guidelines,refining evaluation methods,advancing software and hardware,and enhancing knowledge dissemination,technical training and other actions related to guiding the practical implementation thereby facilitating the wider adoption of PBRTQC.

Guided by the theories of yin-yang and collateral disease， this paper identifies the dysregulation of yang qi ascent and descent as the core pathomechanism of hypertension. Based on clinical experience， a treatment approach centered on the method of raising yang and promoting descent was proposed. Clinically， three major syndrome types were identified. Firstly， deficiency of zong qi （ancestral qi） with blood stasis， obstruction of phlegm-turbidity and blood stasis， and hyperactivity of liver yang. Corresponding empirical formulation， Yizong Huoxue Decoction （益宗活血汤） was applied to tonify zong qi， invigorate blood， and raise yang. Secondly， Lizong Huoxue Decoction （理宗活血汤） was used to resolve phlegm， promote yang qi circulation， and regulate qi and blood. Thirdly， Qinggan Tongluo Decoction （清肝通络饮） was used to clear the liver， dredge collaterals， and subdue hyperactive yang. For special types such as non-dipper hypertension， time-specific syndrome differentiation and treatment can be applied based on a thorough understanding of the underlying pathomechanism， aiming to provide new insights into clinical diagnosis and treatment of hypertension.

Objective To report cases of checkpoint inhibitor pneumonitis （CIP） in patients with advanced biliary tract cancer, aiming to provide additional approaches for the assessment, treatment, and monitoring of this condition. Methods Three patients developed oxygen desaturation and interstitial lung lesions during chemotherapy combined with immunotherapy, and were diagnosed with CIP in collaboration with the respiratory department. Antitumor therapy was discontinued in the acute phase, and glucocorticoids were administered, with regular monitoring of disease progression. During follow-up, case 1 developed lung metastasis; case 2 showed improvement; case 3 had concurrent infection and tumor progression. Results Glucocorticoids improved lung lesions and hypoxic symptoms in patients with CIP, but attention should be paid to the potential for concurrent infections and tumor progression. Conclusions Comprehensive assessment and early identification of CIP are crucial for patients with advanced biliary tract cancer. For those with recurrent symptoms after glucocorticoid therapy, timely and accurate adjustment of the treatment regimen is essential.