ObjectiveTo investigate the role and potential mechanism of microRNA-544 （miRNA-544） in lipopolysaccharide （LPS）-induced liver injury in mice with sepsis， and to provide a new target for the treatment of liver injury in sepsis. MethodsA total of 40 C57BL/6J mice were randomly divided into control group （intraperitoneal injection of normal saline）， model group （intraperitoneal injection of LPS at a dose of 5 mg/kg）， agonist group （intraperitoneal injection of LPS and miR-544 agonist at a dose of 5 mg/kg）， and miR-544 inhibitor group （intraperitoneal injection of LPS and miR-544 inhibitor at a dose of 5 mg/kg）， with 10 mice in each group. An automatic biochemical analyzer was used to measure the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and total bilirubin （TBil） in serum and the liver； Western blot was used to measure the expression levels of monocyte chemotactic protein-1 （MCP-1）， CD16/32， and proteins associated with the NF-κB signaling pathway in the liver； q-PCR and ELISA were used to measure the expression levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） in serum. A one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group， the model group of LPS-induced sepsis had a significant reduction in the expression level of miR-544 in serum and liver tissue （P0.01）， significant pathological changes of the liver （such as inflammatory cell infiltration and central vein congestion）， and significant increases in the levels of liver injury markers （ALT， AST， and TBil） in serum and the liver （all P0.001）， the expression levels of inflammatory factors （MCP-1， CD16/32， TNF-α， IL-6， and IL-1β） （all P0.01）， and the phosphorylation levels of key proteins of the NF-κB pathway （p-IKK， p-I-NF-κ， and p-p65） （all P0.01）. Compared with the model group， the miR-544 inhibitor group had a significant reduction in the expression level of miR-544 in serum and liver tissue （P0.01）， aggravated pathological changes of the liver， and significant increases in the levels of liver injury markers and inflammatory factors （ALT， AST， TBil， MCP-1， CD16/32， TNF-α， IL-6， and IL-1） （all P0.05）， as well as significant increases in the phosphorylation levels of key proteins of the NF-κB pathway （p-IKK， p-I-κB-α， and p-p65） （all P0.01）. On the contrary， the miR-544 agonist group had a significant increase in the expression level of miR-544 in serum and liver tissue （P0.01）， significant alleviation of liver pathological changes， and significant reductions in the levels of liver injury markers and inflammatory factors （ALT， AST， TBil， MCP-1， CD16/32， TNF-α， IL-6， and IL-1β） （all P0.05）， as well as significant reductions in the phosphorylation levels of key proteins of the NF-κB pathway （p-IKK， p-I-κB-α， and p-p65） （all P0.05）. ConclusionThis study shows that miR-544 can alleviate LPS-induced liver injury in mice with sepsis by inhibiting the expression of inflammatory-related proteins and the activation of the NF-κB signaling pathway.

Gastric cancer (GC) is a globally prevalent malignancy with a particularly heavy burden in China. Helicobacter pylori ( H. pylori ) is a Group I carcinogen for GC, with a higher seroprevalence rate indicating a higher GC incidence. However, only approximately 3% of the individuals with H. pylori infection eventually develop GC, and about 2.6% still progress to GC even 10-20 years after the eradication of H. pylori . Thus, the pathogenic mechanism of H. pylori for GC must be elucidated, and high-risk individuals precisely identified. Furthermore, GC can occur even in individuals who have never been infected with H. pylori . As H. pylori infection rates decline, the proportion of H. pylori -negative GC cases is increasing annually, gaining significant research attention. In this review, potential pathogenic mechanisms of H. pylori infection are explored from the aspects of H. pylori virulence factors and host factors (genetic susceptibility and immune microenvironment). Possible risk factors for H. pylori -negative GC include infections by other microorganisms (e.g., bacteria, fungi, and viruses), autoimmune gastritis, bile reflux, genetic mutations, and environmental factors. We aim to review the potential mechanisms for GC with varying H. pylori infection statuses, identify the high-risk individuals, and pose questions that need to be addressed. In the future, as the prevalence of H. pylori infection gradually decreases, GC prevention and management must evolve to address host-specific factors and the growing challenge of H. pylori -negative GC by integrating multidisciplinary perspectives.
Stomach Neoplasms/genetics* ; Humans ; Helicobacter Infections/complications* ; Helicobacter pylori/pathogenicity* ; Risk Factors

OBJECTIVE: To explore the changes of CKLF-like MARVEL transmembrane domain-containing 6 (CMTM6) and programmed death-ligand 1 (PD-L1) expression in gastric mucosal epithelial cells after Helicobacter pylori infection and the regulation of CMTM6 on PD-L1, and to analyze the mRNA expression differences before and after CMTM6 gene knock-out in helicobacter pylori infected gastric epithelial cells by microarray analysis. METHODS: The standard Helicobacter pylori strain ATCC 26695 was co-cultured with human gastric epithelial cell GES-1 for 6, 24 and 48 hours, and the mRNA and protein levels of CMTM6 and PD-L1 were detected by real-time quantitative PCR and Western blot. Using CRISPR/Cas9 to construct CMTM6 gene knockout plasmid and knockout CMTM6 gene of GES-1 cells. Helicobacter pylori was co-cultured with CMTM6 gene knockout and wild type GES-1 cells for 48 hours to detect PD-L1 transcription and protein level changes, and CMTM6 gene knockout GES-1 cells were treated with the proteasome inhibitor MG-132 to detect the changes in PD-L1 protein levels. Agilent Human ceRNA Microarray 2019 was used to detect the differentially expressed genes in CMTM6 gene knockout and wild-type GES-1 cells co-cultured with Hp for 48 hours, and the signal pathway of differentially expressed genes enrichment was analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) database. RESULTS: The mRNA and protein levels of CMTM6 and PD-L1 in GES-1 cells were significantly up-regulated after Helicobacter pylori infection, and CMTM6 mRNA was most significantly up-regulated 48 hours after infection. After CMTM6 gene knockout, the CD274 gene transcription level of Helicobacter pylori infected GES-1 cells did not change significantly, but PD-L1 protein level was significantly down-regulated, and the PD-L1 level increased after the application of proteasome inhibitor MG-132. After CMTM6 gene knockout, 67 genes had more than two times of differential expression. The transcription levels of TMEM68, FERMT3, GPR142, ATP6V1FNB, NOV, UBE2S and other genes were significantly down-regulated. The transcription levels of PCDHGA6, CAMKMT, PDIA2, NTRK3, SPOCK1 and other genes were significantly up-regulated. After CMTM6 gene knockout, ubiquitin-conjugating enzyme E2S (UBE2S) gene expression was significantly down-regulated, which might affect protein ubiquitination degradation. After CMTM6 gene knockout, adrenoceptor alpha 1B (ADRA1B), cholinergic receptor muscarinic 1 (M1), CHRM1, platelet activating factor receptor (PTAFR) gene expression was significantly up-regulated. CONCLUSION Helicobacter pylori infection up-regulates the expression level of CMTM6 in gastric mucosa cells, and CMTM6 can stabilize PD-L1 and maintain the protein level of PD-L1. CMTM6 gene knockout may affect biological behaviors such as protein ubiquitination and cell surface receptor expression.
Humans ; MARVEL Domain-Containing Proteins/metabolism* ; Helicobacter pylori/physiology* ; B7-H1 Antigen/genetics* ; Helicobacter Infections/metabolism* ; Epithelial Cells/metabolism* ; Gastric Mucosa/metabolism* ; Chemokines/metabolism* ; Cell Line ; Gene Knockout Techniques ; Myelin Proteins

Recent studies have found that chondrocytes not only represent the cells targeted by rheumatoid ar-thritis(RA)but may also themselves induce the disease.This dual role indicates the significance of chondrocytes in the pathology of RA.This article summarizes the important roles played by chondrocytes in RA,providing a reference for the pathogenesis and treatment of RA.

Objective:To evaluate the long-term clinical efficacy of endoscopic submucosal dissection (ESD) in the treatment of early colorectal cancer and precancerous lesions, and to explore the risk factors of local recurrence after operation.Methods:From January 1, 2011 to December 31, 2022, the clinical and endoscopic follow-up data of 1 095 patients (1 186 lesions) who underwent ESD at Peking University Third Hospital and were pathologically diagnosed as early colorectal cancer or precancerous lesions after ESD were retrospectively analyzed. The long-term efficacy of ESD was evaluated, which included 5-year overall survival rate, disease-specific survival rate, local recurrence rate, and recurrence-free survival rate. The Kaplan-Meier method was used for survival analysis. Multivariate Cox proportional hazards regression models were performed to analyze local recurrence related clinical pathological factors.Results:After ESD, 1 067 patients were followed up, and the median follow-up period was 44.4 (20.3, 62.1) months. There were 734 patients having endoscopic follow-up (798 lesions, the follow-up rate was 67.0%). During the follow-up period, 26 patients died, and the 5-year overall survival rate and disease-specific survival rate were 96.0% and 100.0%, respectively. During the follow-up period, local recurrence was observed in 17 lesions, with a recurrence rate of 2.1%(17/798) and a median time of recurrence after ESD was 11.8 (4.9, 21.4) months. The 5-year cumulative recurrence rate was 3.4%, and the 5-year recurrence-free survival rate was 94.0%. The results of multivariate Cox regression analysis showed that lesions located in the rectum ( HR=2.64, 95% confidence interval (95% CI) 1.00 to 6.94, P=0.049), histologically incomplete resection ( HR = 4.40, 95% CI 1.62 to 11.94, P=0.004), and positive vertical margin ( HR=10.27, 95% CI 2.95 to 35.77, P<0.001) were independent risk factors for local recurrence after ESD in the treatment of colorectal mucosal lesions. Conclusions:Long-term efficacy of ESD in the treatment of colorectal mucosal lesions are favorable. Lesions located in the rectum, histologically incomplete resection, and positive vertical margin are independent risk factors for local recurrence after ESD in the treatment of colorectal mucosal lesions.

Objective:To evaluate the clinical efficacy of Xiaoqinglong Decoction Granules in the treatment of chronic persistent cold syndrome of bronchial asthma; To explore its treatment mechanism.Methods:A randomized double-blind controlled study was performed. Totally 60 patients from the Respiratory Department of Guang'anmen Hospital, Chinese Academy of Traditional Chinese Medicine from January 2021 to January 2022 were selected as the observation subjects. They were divided into two groups using a random number table method, with 30 cases in each group. The control group was given conventional treatment plus placebo, and the experimental group was given conventional treatment plus Xiaoqinglong Decoction Granules. The treatment for both group lasted for 14d. TCM syndromes and clinical symptoms before and after treatment were scored. Asthma Control Test Questionnaire (ACT) was used to evaluate asthma control status, and the Asthma Quality of Life Questionnaire (Mini AQLQ) was used to evaluate the physiological and psychological effects of asthma on patients; FEV1 was detected using a German Jaeger lung function instrument FEV1/FVC. A exhaled nitric oxide (FeNO) detection instrument was used to observe the changes in FeNO at a flow rate of 50 ml/s, and non-targeted metabolomics analysis was performed using liquid chromatography-mass spectrometry (LC-MS); adverse reactions were observed during treatment and drug safety was evaluated.Results:Eventually 47 cases were included, 24 cases of test group and of 23 cases of control group. Xiaoqinglong Decoction Granules could reduce the TCM syndrome score of patients with chronic duration cold syndrome of asthma ( P<0.05). 2 weeks after treatment, follow up for 4 weeks experimental group clinical symptom score [3.00(1.00,4.00),3.00(0.00,4.00) vs. 3.5(3.00,5.00), Z=8.62], breathing symptom scores [1.00(0.00,1.00),1.00(0.00,1.00) vs. 1.00(0.75,2.00), Z=6.80], cough symptom score [0.50(0.00,1.00),1.00(0.00,1.00) vs. 1.00(0.00,1.25), Z=6.12] were lower than those in the experimental group before treatment in the same group ( P<0.01 or P<0.05). The ACT score of the experimental group at 4 weeks of follow-up was [22.50 (21.00, 24.00) vs. 9.00 (15.00, 21.50), Z=-4.87], Mini AQLQ score (78.5 ± 12.46 vs. 71.27 ± 9.70, t=-2.46) and the control group had an ACT score of [24.00 (19.00, 25.00) vs. 21.5 (8.00, 23.00) Z=-3.18] at 4 weeks of follow-up was higher than before treatment in the same group ( P<0.01 or P<0.05). The experimental group was followed up for 4 weeks with a FEV1 of [2.96 (2.27, 3.49) L vs. 2.60 (2.32, 3.49) L, Z=-3.72], FEV1/FVC [(80.83 ± 6.84)% vs. (77.46 ± 8.15)%, t=-2.32] and FeNO [24.00 (12.50, 31.00) ppb vs. 30.00 (17.00, 91.00) ppb, Z=-3.72] was higher than before treatment in the same group ( P<0.01 or P<0.05). Through LC-MS technique analysis, there were 75 kinds of different metabolites between the experimental group before and after treatment, and 295 kinds of different metabolites between the control group and the experimental group after treatment. Further intersection of differential metabolites showed that they were mainly concentrated in histidine metabolic pathway, phosphonate metabolic pathway and phosphate metabolic pathway. Related metabolites 2-aminoethyl phosphonate and thiomalonic acid were involved. Conclusions:Xiaoqinglong Decoction Granules can effectively improve the TCM syndrome and clinical symptoms of patients with chronic persistent cold syndrome of asthma, especially for wheezing, cough and chest tightness, which can improve the levels of FEV1 and FEV1/FVC in patients and effectively reduce FeNO. Through metabolomics studies, it is speculated that Xiaoqinglong Decoction Granules may play a role in the treatment of asthma by regulating histidine metabolism pathway through thiomalonic acid.

Objective To analyze the clinical,pathological,and endoscopic features of differentiated early gastric cancer,and to study predictors to identify and screen high risk patients with early gastric cancer submucosal infiltration.Methods A total of 172 patients with differentiated early gastric cancer treated by surgical or endoscopic submucosal dissection in our hospital from January 2017 to December 2022 were included,which were divided into the mucosal layer group(144 patients)and submucosal layer group(28 patients)based on postoperative pathology.The clinical,pathological,and white-light endoscopy(WLE)and linked color imaging(LCI)features of the 2 groups were compared.The color difference between the lesion and the surrounding mucosa was evaluated by using the Commission International de L'Eclairage(CIE)L*a*b*system.Indicators with significant differences were included to multifactor logistic stepwise regression analysis(forward method)for the identification and screening of predictors.Results A history of alcohol consumption(P=0.037),a history of smoking(P=0.035),thickening of the gastric wall on enhanced CT(P=0.032),a lesion located in the upper 1/3(P＜0.001)or middle 1/3(P=0.009)part of the stomach,depressed macroscopic type(P＜0.001),marked margin elevation(P=0.003),presence of fold changes(P=0.006),color difference ≥12.3 under WLE(P=0.003)and≥18.2 under LCI(P=0.002)were associated with submucosal infiltration.Multivariate analysis showed that lesions located in the upper 2/3 portion of the stomach(OR=5.463,95％CI:2.562-11.648,P＜0.001),depressed macroscopic type(OR=5.992,95％CI:1.624-22.100,P=0.007),marked margin elevation(OR=4.338,95％CI:1.124-16.747,P=0.033),and color difference ≥18.2 under LCI(OR=4.675,95％CI:1.342-16.288,P=0.015)were independent risk factors for infiltration of submucosal layer of lesions.Conclusion Lesions with depressed macroscopic type,marked elevated margins,located in the upper 2/3 part of the stomach,and having a large color difference from the surrounding mucosa under LCI are high-risk lesions for submucosal infiltration and require more aggressive intervention.

Background:Cronkhite Canada syndrome(CCS)is a rare non hereditary disease of unknown etiology,there are still challenges in its diagnosis and treatment.Aims:To explore the clinical characteristics and treatment responses of CCS in Chinese population,and to improve the understanding of its diagnosis and treatment.Method:Retrospective analysis and summary of clinical data of CCS patients at Peking University Third Hospital from 2012 to 2022.Results:From the clinical data of a total of 9 cases,the patients'average age was 63.89 years,and the male to female ratio was 1.25.all patients presented with multiple gastrointestinal polyps,non-specific gastrointestinal symptoms and at least one ectodermal manifestation.Laboratory tests for fecal occult blood were positive,and iron deficiency anemia and hypoalbuminemia were common.Histological manifestations were hyperplastic polyps and adenomatous polyps,with commonly found eosinophil infiltration.Patients'conditions could be complicated with severe osteoporosis and fractures,intestinal bacterial overgrowth,asthma,membranous nephropathy,and nodular goiter.Most patients obtained positive currative effect with glucocorticoids therapy,but recurrence of the disease may occur during or after hormone reduction.Azathioprine treatment was attempted in one patient,but the effect was poor.Conclusions:The etiology and pathogenesis of CCS are unclear,and glucocorticoids therapy is still the main treatment method.However,there are still challenges for patients with glucocorticoids resistance and contraindications to glucocorticoids therapy.

Objective:To explore the predictive value of lipoprotein-associated phospholipase A2 (Lp-PLA2) combined with systemic immune-inflammation index (SII) for coronary heart disease risk in patients with type 2 diabetes mellitus.Methods:A prospective study was performed, 130 patients with type 2 diabetes mellitus from May 2018 to May 2021 in the People′s Hospital of Pengzhou were selected. All patients underwent coronary angiography examination, and 49 cases were complicated with coronary heart disease (coronary heart disease group), 81 cases were not complicated with coronary heart disease (non-coronary heart disease group). The clinical data were recorded; the white blood cell, monocyte, platelet, neutrophils, lymphocyte, hemoglobin, total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), fasting blood glucose, C-reactive protein (CRP), glycated hemoglobin and Lp-PLA2 were detected; and the SII was calculated. Multivariate Logistic regression analysis was used to analyze the independent risk factors of coronary heart disease in patients with type 2 diabetes mellitus; the efficacy of Lp-PLA2 and SII in predicting the coronary heart disease in patients with type 2 diabetes mellitus was evaluated using the receiver operating characteristics (ROC) curve.Results:There were no statistical differences in white blood cell, hemoglobin, TC, TG, LDL-C, fasting blood glucose, glycated hemoglobin and CRP between two groups ( P>0.05); the monocyte, platelet, neutrophils, Lp-PLA2 and SII in coronary heart disease group were significantly higher than those in the non-coronary heart disease group: (0.55 ± 0.22) × 10 9/L vs. (0.40 ± 0.11) × 10 9/L, (227.84 ± 40.76) × 10 9/L vs. (205.81 ± 39.04) × 10 9/L, (6.78 ± 1.45) × 10 9/L vs. (6.30 ± 1.18) × 10 9/L, (240.67 ± 41.48) μg/L vs. (214.83 ± 36.35) μg/L and 1 245.76 ± 383.08 vs. 929.84 ± 260.27, the lymphocyte and HDL-C were significantly lower than those in the non-coronary heart disease group: (1.26 ± 0.17) × 10 9/L vs. (1.41 ± 0.19) × 10 9/L and (1.15 ± 0.14) mmol/L vs. (1.23 ± 0.21) mmol/L, and there were statistical differences ( P<0.01 or <0.05). Multivariate Logistic regression analysis result showed that monocyte, HDL-C, Lp-PLA2 and SII were the independent risk factors of coronary heart disease in patients with type 2 diabetes mellitus ( OR = 1.812, 1.013, 1.013 and 2.004; 95% CI 4.430 to 6.801, 0.992 to 1.013, 1.001 to 1.026 and 0.004 to 0.855; P<0.01 ore <0.05). ROC curve analysis result showed that the area under the curve of Lp-PLA2 combined with SII to predict the coronary heart disease in patients with type 2 diabetes mellitus was significantly larger than Lp-PLA2 and SII alone: 0.783 (95% CI 0.702 to 0.851) vs. 0.681 (95% CI 0.593 to 0.760) and 0.744 (95% CI 0.660 to 0.816), and there was statistical difference ( P<0.05). Conclusions:Lp-PLA2 and SII are independent risk factors for coronary heart disease in patients with type 2 diabetes mellitus, and their combined detection can improve the predictive value of coronary heart disease.

Objective:To study the effect of β-amyloid(Aβ) on calcium homeostasis and endoplasmic reticulum calcium storage of hippocampal neurons in rats.Methods:A total of 60 adult male SD rats were randomly divided into six groups by body mass matching method, with 10 rats in each group.Three groups were injected Aβ 25-35 into the hippocampus(2 μL per side), and divided into low dose group(2.5 μg/μL), medium dose group(5.0 μg/μL) and high dose group(7.5 μg/μL) respectively, and the other 3 groups were set up as the normal saline group(2 μL 0.9% sodium chloride solution), sham-operated group(rats craniotomy without injection) and normal control group(normal feeding without any treatment). The rats were fed until 14 days after operation, and the behavior and state of the rats were observed and recorded, as well as the body weight and total food intake ratio.And the rat hippocampal cells endoplasmic reticulum pathological change were observed by using the electron microscope and light microscope, meanwhile the concentration of intracellular free Ca 2+ ions was detected by the laser scanning confocal microscope and the expression level of PS, SERCA and RyR mRNA and protein by real-time PCR and Western blot methods respectively.The experimental results were analyzed using SPSS 25.0 software for statistical analysis. Repeated measurement ANOVA and one-way ANOVA were used for multi-group comparison, and Dunnett test and Tukey test were used for further pairwise comparison. Results:(1) The body weight of rats in each group was analyzed by repeated measurement ANOVA, and the difference of time effect was statistically significant( F=153.15, P<0.001), but there was no significant difference between the intergroup effects and interaction effects( F=1.547, P=0.374; F=1.598, P=0.113). The body weight of high, medium and low dose Aβ 25-35 groups at 7, 14 days after injection had no significant difference compared with the control group(all P>0.05). The food utilization rates of the high, medium and low doses of Aβ 25-35 groups were(22.9±4.0)%, (23.0±4.2)% and(22.6±3.2)%, respectively, and there was no significant difference compared with the control group((23.7±5.0)%, P>0.05). Within 14 days after injection, listlessness and lethargy were observed in rats in the high dose Aβ 25-35 group.(2) Pathological observation results showed that the endoplasmic reticulum of rat hippocampal cells in the high dose and medium dose groups of Aβ 25-35 was expanded and swelled, and the mitochondria were swollen and deformed.(3) 14 days after Aβ 25-35 injection, the fluorescence intensity of free calcium in hippocampus of rats in high, medium and low dose groups were(820.43±6.89), (720.12±4.30) and (680.50±4.32), respectively, which were all higher than that in the control group(592.17±3.97)(all P<0.001). (4) RT-PCR and Western blot results showed that compared with the control group, high dose and medium dose Aβ 25-35 injection could up-regulate the expression of PS and SERCA mRNA and protein in hippocampal cells(all P<0.05), while down-regulate the expression of RyR mRNA and protein in hippocampal cells(all P<0.05). Conclusion:The deposition of Aβ 25-35 in hippocampal tissues can disrupt the homeostasis of calcium ions in hippocampal tissues, and then cause the increase of free calcium and its related proteins, thus playing the neurotoxic role.