BACKGROUND: Arsenic trioxide (ATO) is indicated as a broad-spectrum medicine for a variety of diseases, including cancer and cardiac disease. While the role of ATO in hepatic ischemia/reperfusion injury (HIRI) has not been reported. Thus, the purpose of this study was to identify the effects of ATO on HIRI. METHODS: In the present study, we established a 70% hepatic warm I/R injury and partial hepatectomy (30% resection) animal models in vivo and hepatocytes anoxia/reoxygenation (A/R) models in vitro with ATO pretreatment and further assessed liver function by histopathologic changes, enzyme-linked immunosorbent assay, cell counting kit-8, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Small interfering RNA (siRNA) for extracellular signal-regulated kinase (ERK) 1/2 was transfected to evaluate the role of ERK1/2 pathway during HIRI, followed by ATO pretreatment. The dynamic process of autophagic flux and numbers of autophagosomes were detected by green fluorescent protein-monomeric red fluorescent protein-LC3 (GFP-mRFP-LC3) staining and transmission electron microscopy. RESULTS: A low dose of ATO (0.75 μmol/L in vitro and 1 mg/kg in vivo ) significantly reduced tissue necrosis, inflammatory infiltration, and hepatocyte apoptosis during the process of hepatic I/R. Meanwhile, ATO obviously promoted the ability of cell proliferation and liver regeneration. Mechanistically, in vitro  studies have shown that nontoxic concentrations of ATO can activate both ERK and phosphoinositide 3-kinase-serine/threonine kinase (PI3K-AKT) pathways and further induce autophagy. The hepatoprotective mechanism of ATO, at least in part, relies on the effects of ATO on the activation of autophagy, which is ERK-dependent. CONCLUSION Low, non-toxic doses of ATO can activate ERK/PI3K-AKT pathways and induce ERK-dependent autophagy in hepatocytes, protecting liver against I/R injury and accelerating hepatocyte regeneration after partial hepatectomy.
Animals ; Arsenic Trioxide ; Autophagy/physiology* ; Reperfusion Injury/prevention & control* ; Mice ; Male ; Proto-Oncogene Proteins c-akt/physiology* ; Arsenicals/therapeutic use* ; Oxides/therapeutic use* ; Liver/metabolism* ; Extracellular Signal-Regulated MAP Kinases/metabolism* ; Mice, Inbred C57BL

Objective:To investigate the effects of Vibrio vulnificus ( Vv) quorum-sensing protein LuxS on the homeostasis of pulmonary innate immune cells in sepsis-induced acute lung injury. Methods:This study constructed luxS knockout and complemented Vv strains. The time required for wild type, luxS knockout, and complemented Vv strains to grow to an absorbance of 600 nm in liquid medium was measured using a spectrophotometer. Iron-overloaded mice were intraperitoneally infected with 1×10 5 CFU of the above three kinds of Vv strains, respectively. Clinical scoring for sepsis-induced dyspnea was used to evaluate the respiratory quality in mice. At 7 h after infection, the pathological changes in lung tissues were observed by HE staining; the bacterial loads in lung tissues were measured; the single-cell suspension of lung tissues was analyzed by flow cytometry. Uniform manifold approximation and projection (UMAP) was used to reduce the dimension of the distribution of CD45 + immune cells in lung tissues of mice in the PBS control group and infection groups with different strains. The frequency and absolute number of innate immune cells in lung tissues were analyzed by multicolor flow cytometry. One-way analysis of variance and t test were used for statistical analysis. Results:There was no significant difference in the growth rate of wild type, luxS knockout, and complemented Vv strains in liquid medium. Compared with the mice infected with the wild type or complemented strain, the mice infected with the luxS knockout strain exhibited overall alleviated respiratory difficulty, decreased inflammatory cell infiltration in lung tissues, and reduced bacterial load in lung tissues ( P<0.05). Besides, there was no significant difference in clinical respiratory scores, inflammatory cell infiltration, or bacterial loads between the mice infected with the complemented strain and wild type strain. UMAP analysis showed that compared with the mice infected with the luxS knockout strain, the mice infected with the wild type or complemented strain showed increased proportions of neutrophils and eosinophils in lung tissues. Results of multicolor flow cytometry analysis further verified that the proportions of neutrophils and eosinophils were significantly lower in the mice infected with the luxS knockout strain than in the mice infected with wild type or complemented strain ( P<0.01, P<0.000 1), while the proportion of alveolar macrophages was significantly higher as compared with that in the mice infected with wild type or complemented strain ( P<0.01). Conclusion:During Vv infection, LuxS may promote acute lung injury by affecting the homeostasis of neutrophils, eosinophils and resident macrophages in lung tissues.

Objective:To investigate the role of autophagy in the regulatory of phagocytosis in Vibrio vulnificus ( V. vulnificus)-infected murine macrophages. Methods:The expression of cellular autophagy-related proteins in PBS-treated and V. vulnificus-infected RAW264.7 and BMMφ cells was detected by Western blot. The co-localization of V. vulnificus-GFP and LC3Ⅱ protein in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells were detected using confocal microscopy. The phagocytosis of V. vulnificus in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells with or without autophagy inhibition using Bafilomycin A1 was detected by flow cytometry. Results:The up-regulated levels of Atg7, Atg12 and Atg16L1 proteins, increased LC3Ⅱ/actin ratio, as well as down-regulated p62 protein levels were significantly detected in V. vulnificus-infected RAW264.7 and BMMφ cells. The co-localization of V. vulnificus-GFP and LC3Ⅱ protein was clearly observed in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells. Enhanced phagocytosis of V. vulnificus and increased autophagy were exhibited in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells, while weakened phagocytosis, accumulation of Atg7, Atg12, Atg16L1, LC3Ⅱ and p62 protein levels, as well as blocking autophagy flux were detected in those cells within autophagy inhibition using Bafilomycin A1. Conclusion:Autophagy induced by V. vulnificus infection could promote phagocytosis of V. vulnificus in macrophages.

To elucidate the genomic characteristics of Salmonella strains derived from sheep,this study employed various methods,including bacterial isolation and identification,biochemical identi-fication,pathogenicity test,whole-genome sequencing,and BLAST comparison,along with the screening of integrative conjugative elements(ICE)using ICEfinder and EasyFig for comparative analysis,as well as plasmid comparisons utilizing PlasmidBrig.The results revealed the isolation of a Gram-negative,non-spore-forming bacillus from nasal swabs of diseased sheep,which formed gray-white,smooth-surfaced,and neatly edged circular colonies on TSA sheep blood agar.On XLT-4 agar medium,it produced smooth-surfaced,white,circular colonies.The bacterium was identified as Salmonella enterica through 16S rRNA sequencing and biochemical identification.This bacteri-um induces hemorrhaging in the intestines of guinea pigs,resulting in their demise within a 48-hour period.The pathogen exhibits high virulence.Whole-genome alignment demonstrated a high degree of homology with Salmonella enterica subsp.diarizonae serotype 61:k:1,5,(7).ICE screen-ing and comparative analysis indicated the presence of a novel ICE in this strain,characterized by a core structural framework that includes an integrative shear module,a mobilizable processing mod-ule,a conjugative pair formation module,and a regulatory module.Notably,ICE from different spe-cies containing the same integrase exhibited identical inverted repeat sequences and insertion sites at tRNAPhe.Plasmid homology comparisons revealed that plasmid sequences from different strains of Salmonella enterica subsp.diarizonae serotype 61:k:1,5,(7)also showed high homology;however,the homology with plasmid sequences from other Salmonella and Escherichia coli strains was only 50％.These findings indicate that the isolated strain is Salmonella enterica subsp.diarizonae serotype 61:k:1,5,(7)and contains a novel ICE as well as a plasmid.This study fur-ther enriches the molecular epidemiology of Salmonella and provides a theoretical basis for the prevention and control of infections caused by this pathogen.

Objective:To investigate the effect of short-term intensive insulin pump therapy on blood glucose control and renal function in type 2 diabetes mellitus (T2DM) patients with diabetic kidney disease (DKD).Methods:The clinical data of 360 T2DM patients with DKD in Shanxi Bethune Hospital from January 2022 to October 2024 were retrospectively analyzed. These patients were divided into an observation group ( n=192; treated with continuous subcutaneous insulin infusion) and a control group ( n=168; treated with conventional hypoglycemic therapy) according to the different treatment modalities; subsequently, 103 pairs of patients entered final analysis by using the propensity score matching method. The differences in glycemic control and improvement of renal function were compared between these two groups. Results:Before the treatment, the blood glucose-, blood glucose fluctuation-, and kidney function-related indicators showed no significant differences between these two groups (all P>0.05). After the treatment, fasting blood glucose, two-hour postprandial blood glucose, and average blood glucose significantly decreased in both groups and were significantly lower in the observation group than in the control group (all P<0.05). In addition, the amplitude of postprandial blood glucose fluctuation, standard deviation of intraday multi-point blood glucose, and difference between the maximum and minimum intraday blood glucose values significantly declined after treatment and were also significantly lower in the observation group than in the control group (all P<0.05). The observation group had a significantly higher total glycemic control rate and a significantly lowr incidence of hypoglycemia (both P<0.05). Finally, the 24-hour urine protein level, serum creatinine level, and urine albumin/creatinine ratio of the observation group significantly dropped after the treatment, and these results were significantly lower than those in the observation group (all P<0.05). Conclusions:Intensive insulin pump therapy benefits T2DM patients with DKD by improving glycemic control and enhancing renal function.

Objective Through a non-inferiority trial design,to verify whether the efficacy of transcuta-neous electrical acupoint stimulation(TEAS)in intervening with nausea and vomiting after painless gastros-copy and colonoscopy was not inferior to that of ondansetron,and evaluate its effect on patient recovery.Meth-ods A total of 112 patients who experienced nausea and vomiting after painless gastroscopy and colonoscopy in the gastroscopy and colonoscopy center of the hospital from March 2022 to March 2023 were selected as the research subjects,and divided into the TEAS group(n=56)and the control group(n=56)according to the random number table method.The TEAS group received TEAS intervention,while the control group received ondansetron hydrochloride injection intervention.The non-inferiority margin was set at 10.0％.The interven-tion effective rate of nausea and vomiting after examination,as well as heart rate,blood pressure,length of stay,satisfaction,and visual analogue score(VAS)were compared between the two groups.Results The in-tervention effective rate of the TEAS group was close to that of the control group[82.4％(46/56)vs.80.4％(45/56)],risk difference(RD)=2.0％(95％CI:-9.8％to 13.8％),the lower limit of 95％CI(-9.8％)was higher than the preset cut-off value(-10.0％),and non-inferiority was established(P=0.032).Com-pared with before the examination,the heart rates of the two groups were slower,and the levels of systolic blood pressure(SEP)and diastolic blood pressure(DBP)were lower during the examination,and the differ-ences were statistically significant(P＜0.05).There was no statistically significant difference in the two groups of heart rate,SBP,DBP,length of stay[(69.9±14.2)min vs.(75.3±22.8)min],satisfaction score(9.8±0.6 vs.9.2±2.0),VAS(1.5±0.5 vs.2.1±0.5,P＞0.05).Conclusion The efficacy of TEAS in in-tervening with nausea and vomiting after painless gastroscopy and colonoscopy is not inferior to that of ondan-setron.

Objective:To investigate the effect of short-term intensive insulin pump therapy on blood glucose control and renal function in type 2 diabetes mellitus (T2DM) patients with diabetic kidney disease (DKD).Methods:The clinical data of 360 T2DM patients with DKD in Shanxi Bethune Hospital from January 2022 to October 2024 were retrospectively analyzed. These patients were divided into an observation group ( n=192; treated with continuous subcutaneous insulin infusion) and a control group ( n=168; treated with conventional hypoglycemic therapy) according to the different treatment modalities; subsequently, 103 pairs of patients entered final analysis by using the propensity score matching method. The differences in glycemic control and improvement of renal function were compared between these two groups. Results:Before the treatment, the blood glucose-, blood glucose fluctuation-, and kidney function-related indicators showed no significant differences between these two groups (all P>0.05). After the treatment, fasting blood glucose, two-hour postprandial blood glucose, and average blood glucose significantly decreased in both groups and were significantly lower in the observation group than in the control group (all P<0.05). In addition, the amplitude of postprandial blood glucose fluctuation, standard deviation of intraday multi-point blood glucose, and difference between the maximum and minimum intraday blood glucose values significantly declined after treatment and were also significantly lower in the observation group than in the control group (all P<0.05). The observation group had a significantly higher total glycemic control rate and a significantly lowr incidence of hypoglycemia (both P<0.05). Finally, the 24-hour urine protein level, serum creatinine level, and urine albumin/creatinine ratio of the observation group significantly dropped after the treatment, and these results were significantly lower than those in the observation group (all P<0.05). Conclusions:Intensive insulin pump therapy benefits T2DM patients with DKD by improving glycemic control and enhancing renal function.

Objective:To investigate the effects of Vibrio vulnificus ( Vv) quorum-sensing protein LuxS on the homeostasis of pulmonary innate immune cells in sepsis-induced acute lung injury. Methods:This study constructed luxS knockout and complemented Vv strains. The time required for wild type, luxS knockout, and complemented Vv strains to grow to an absorbance of 600 nm in liquid medium was measured using a spectrophotometer. Iron-overloaded mice were intraperitoneally infected with 1×10 5 CFU of the above three kinds of Vv strains, respectively. Clinical scoring for sepsis-induced dyspnea was used to evaluate the respiratory quality in mice. At 7 h after infection, the pathological changes in lung tissues were observed by HE staining; the bacterial loads in lung tissues were measured; the single-cell suspension of lung tissues was analyzed by flow cytometry. Uniform manifold approximation and projection (UMAP) was used to reduce the dimension of the distribution of CD45 + immune cells in lung tissues of mice in the PBS control group and infection groups with different strains. The frequency and absolute number of innate immune cells in lung tissues were analyzed by multicolor flow cytometry. One-way analysis of variance and t test were used for statistical analysis. Results:There was no significant difference in the growth rate of wild type, luxS knockout, and complemented Vv strains in liquid medium. Compared with the mice infected with the wild type or complemented strain, the mice infected with the luxS knockout strain exhibited overall alleviated respiratory difficulty, decreased inflammatory cell infiltration in lung tissues, and reduced bacterial load in lung tissues ( P<0.05). Besides, there was no significant difference in clinical respiratory scores, inflammatory cell infiltration, or bacterial loads between the mice infected with the complemented strain and wild type strain. UMAP analysis showed that compared with the mice infected with the luxS knockout strain, the mice infected with the wild type or complemented strain showed increased proportions of neutrophils and eosinophils in lung tissues. Results of multicolor flow cytometry analysis further verified that the proportions of neutrophils and eosinophils were significantly lower in the mice infected with the luxS knockout strain than in the mice infected with wild type or complemented strain ( P<0.01, P<0.000 1), while the proportion of alveolar macrophages was significantly higher as compared with that in the mice infected with wild type or complemented strain ( P<0.01). Conclusion:During Vv infection, LuxS may promote acute lung injury by affecting the homeostasis of neutrophils, eosinophils and resident macrophages in lung tissues.

Objective:To investigate the role of autophagy in the regulatory of phagocytosis in Vibrio vulnificus ( V. vulnificus)-infected murine macrophages. Methods:The expression of cellular autophagy-related proteins in PBS-treated and V. vulnificus-infected RAW264.7 and BMMφ cells was detected by Western blot. The co-localization of V. vulnificus-GFP and LC3Ⅱ protein in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells were detected using confocal microscopy. The phagocytosis of V. vulnificus in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells with or without autophagy inhibition using Bafilomycin A1 was detected by flow cytometry. Results:The up-regulated levels of Atg7, Atg12 and Atg16L1 proteins, increased LC3Ⅱ/actin ratio, as well as down-regulated p62 protein levels were significantly detected in V. vulnificus-infected RAW264.7 and BMMφ cells. The co-localization of V. vulnificus-GFP and LC3Ⅱ protein was clearly observed in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells. Enhanced phagocytosis of V. vulnificus and increased autophagy were exhibited in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells, while weakened phagocytosis, accumulation of Atg7, Atg12, Atg16L1, LC3Ⅱ and p62 protein levels, as well as blocking autophagy flux were detected in those cells within autophagy inhibition using Bafilomycin A1. Conclusion:Autophagy induced by V. vulnificus infection could promote phagocytosis of V. vulnificus in macrophages.

To elucidate the genomic characteristics of Salmonella strains derived from sheep,this study employed various methods,including bacterial isolation and identification,biochemical identi-fication,pathogenicity test,whole-genome sequencing,and BLAST comparison,along with the screening of integrative conjugative elements(ICE)using ICEfinder and EasyFig for comparative analysis,as well as plasmid comparisons utilizing PlasmidBrig.The results revealed the isolation of a Gram-negative,non-spore-forming bacillus from nasal swabs of diseased sheep,which formed gray-white,smooth-surfaced,and neatly edged circular colonies on TSA sheep blood agar.On XLT-4 agar medium,it produced smooth-surfaced,white,circular colonies.The bacterium was identified as Salmonella enterica through 16S rRNA sequencing and biochemical identification.This bacteri-um induces hemorrhaging in the intestines of guinea pigs,resulting in their demise within a 48-hour period.The pathogen exhibits high virulence.Whole-genome alignment demonstrated a high degree of homology with Salmonella enterica subsp.diarizonae serotype 61:k:1,5,(7).ICE screen-ing and comparative analysis indicated the presence of a novel ICE in this strain,characterized by a core structural framework that includes an integrative shear module,a mobilizable processing mod-ule,a conjugative pair formation module,and a regulatory module.Notably,ICE from different spe-cies containing the same integrase exhibited identical inverted repeat sequences and insertion sites at tRNAPhe.Plasmid homology comparisons revealed that plasmid sequences from different strains of Salmonella enterica subsp.diarizonae serotype 61:k:1,5,(7)also showed high homology;however,the homology with plasmid sequences from other Salmonella and Escherichia coli strains was only 50％.These findings indicate that the isolated strain is Salmonella enterica subsp.diarizonae serotype 61:k:1,5,(7)and contains a novel ICE as well as a plasmid.This study fur-ther enriches the molecular epidemiology of Salmonella and provides a theoretical basis for the prevention and control of infections caused by this pathogen.