ObjectiveTo investigate the mechanism of Huazhuo Jiedu prescription in treating ulcerative colitis （UC） by regulating mitophagy. MethodsThe genes related to mitophagy and UC were retrieved from GeneCards， and then the common genes of mitophagy and UC were analyzed by metascape to identify the genes related to mitophagy in UC. Animal experiments were carried out to decipher the mechanism by which Huazhuo Jiedu prescription treated UC by regulating mitophagy. Sixty C57BL/6 male mice were randomized into normal， model， high-， medium-， and low-dose （50， 25， 12.5 g·kg^-1， respectively） Huazhuo Jiedu prescription， and mesalazine （0.52 g·kg^-1·d^-1） groups， with 10 mice in each group. After successful modeling by the dextran sulfate sodium-free drinking method， the colonic mucosal damage was observed by hematoxylin-eosin staining， and the ultracellular structure of colon mucosa was observed by transmission electron microscopy. The expression levels of mitophagy-related proteins PTEN-induced putative kinase 1 （PINK1） and Parkin protein were determined by Western blot. The expression of prohibitin 2 （PHB2）， ubiquitin-specific protease 15 （USP15）， ubiquitin-specific protease 30 （USP30） in the colon tissue was detected by immunofluorescence （IF）. ResultsAll the drug intervention groups showed ameliorated pathological manifestations of the colonic mucosa and improved mitochondrial structures in UC mice. Compared with the normal group， the model group demonstrated up-regulated protein levels of PINK1 and Parkin （P<0.05）， enhanced average fluorescence intensity of PHB2 （P<0.05）， and weakened average fluorescence intensity of USP15 and USP30 （P<0.05）. Compared with the model group， the mesalazine group and the high- and medium-dose Huazhuo Jiedu prescription groups showcased down-regulated protein levels of PINK1 and Parkin （P<0.05）， decreased average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. The low-dose Huazhuo Jiedu prescription group showed down-regulated protein levels of PINK1 and Parkin （P<0.05）， weakened average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. ConclusionHuazhuo Jiedu prescription can attenuate the intestinal mucosal injury and improve the mitochondrial cell ultrastructure in UC mice by regulating the expression of PINK1-Parkin pathway and inhibiting excessive mitophagy.

ObjectiveTo investigate the mechanism of Huazhuo Jiedu prescription in treating ulcerative colitis （UC） by regulating mitophagy. MethodsThe genes related to mitophagy and UC were retrieved from GeneCards， and then the common genes of mitophagy and UC were analyzed by metascape to identify the genes related to mitophagy in UC. Animal experiments were carried out to decipher the mechanism by which Huazhuo Jiedu prescription treated UC by regulating mitophagy. Sixty C57BL/6 male mice were randomized into normal， model， high-， medium-， and low-dose （50， 25， 12.5 g·kg^-1， respectively） Huazhuo Jiedu prescription， and mesalazine （0.52 g·kg^-1·d^-1） groups， with 10 mice in each group. After successful modeling by the dextran sulfate sodium-free drinking method， the colonic mucosal damage was observed by hematoxylin-eosin staining， and the ultracellular structure of colon mucosa was observed by transmission electron microscopy. The expression levels of mitophagy-related proteins PTEN-induced putative kinase 1 （PINK1） and Parkin protein were determined by Western blot. The expression of prohibitin 2 （PHB2）， ubiquitin-specific protease 15 （USP15）， ubiquitin-specific protease 30 （USP30） in the colon tissue was detected by immunofluorescence （IF）. ResultsAll the drug intervention groups showed ameliorated pathological manifestations of the colonic mucosa and improved mitochondrial structures in UC mice. Compared with the normal group， the model group demonstrated up-regulated protein levels of PINK1 and Parkin （P<0.05）， enhanced average fluorescence intensity of PHB2 （P<0.05）， and weakened average fluorescence intensity of USP15 and USP30 （P<0.05）. Compared with the model group， the mesalazine group and the high- and medium-dose Huazhuo Jiedu prescription groups showcased down-regulated protein levels of PINK1 and Parkin （P<0.05）， decreased average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. The low-dose Huazhuo Jiedu prescription group showed down-regulated protein levels of PINK1 and Parkin （P<0.05）， weakened average fluorescence intensity of PHB2 （P<0.05）， and enhanced average fluorescence intensity of USP15 and USP30 （P<0.05）. ConclusionHuazhuo Jiedu prescription can attenuate the intestinal mucosal injury and improve the mitochondrial cell ultrastructure in UC mice by regulating the expression of PINK1-Parkin pathway and inhibiting excessive mitophagy.

ObjectiveTo explore the therapeutic effect and mechanism of Xianglian Huazhuo prescription on chronic atrophic gastritis （CAG） in rats based on the Hedgehog signaling pathway. MethodsThe CAG rat model was established by sodium salicylate， N-methyl-N′-nitro-N-nitroguanidine （MNNG）， and irregular feeding. The successfully modeled rats were randomly divided into a model group （180 mg·L^-1）， a moradan group （1.4 g·kg^-1）， and Xianglian Huazhuo Prescription groups with high， medium， and low doses （36， 9， 18 g·kg^-1）， followed by drug intervention. Hematoxylin-eosin （HE） staining was used to observe morphological changes in the gastric mucosa. Transmission electron microscopy was used to observe the ultrastructure of gastric mucosa cells. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of Sonic Hedgehog （Shh）， Patched 1 （Ptch1）， and Glioma-associated oncogene homolog 1 （Gli1）. Western blot was used to detect the protein expression levels of Shh， Ptch1， and Gli1 in the gastric mucosa. Immunohistochemistry was used to observe the protein expression of the epithelial marker E-cadherin. ResultsCompared with the normal group， the CAG model group showed a reduction in gastric mucosal intrinsic glands and infiltration of inflammatory cells. The ultrastructure of gastric mucosal cells showed nuclear pyknosis， fewer mitochondria， and abnormal mitochondrial structure. The mRNA and protein expression of Shh， Ptch1， and Gli1 in the gastric mucosa were significantly decreased （P<0.05）， and E-cadherin protein expression was decreased. Compared with the model group， the intervention groups showed varying degrees of improvement in histopathological morphology and cellular ultrastructure. The mRNA and protein expression of Shh， Ptch1， Gli1， and E-cadherin increased to varying degrees. Xianglian Huazhuo Prescription upregulated the expression of key Hedgehog pathway factors and E-cadherin at both the mRNA and protein levels （P<0.05）. ConclusionXianglian Huazhuo prescription has a therapeutic effect on CAG in rats， and its mechanism may be related to activation of the Hedgehog signaling pathway and inhibition of epithelial-mesenchymal transition （EMT）.

Increasing evidence has underscored the significance of post-stroke alterations along gut-brain axis, while its role in pathogenesis and treatment of ischemic stroke (IS) remains largely unexplored. This study aimed to elucidate the therapeutic effects and action targets of Panax notoginseng saponins (PNS) on IS and explore a novel pathogenesis and treatment strategy of IS via profiling the microbial community and metabolic characteristics along gut-brain axis. Our findings revealed for the first time that the therapeutic effect of PNS on IS was microbiota-dependent. Ischemia/reperfusion (I/R) modeling significantly down-regulated Lactobacilli in rats, and PNS markedly recovered Lactobacilli, particularly Lactobacillus reuteri (L.Reu). Metabolomics showed a significant reduction in serum histidine (HIS) in clinical obsolete IS patients and rehabilitation period I/R rats. Meanwhile, the L.Reu colonization in I/R rats exhibited significant neuroprotective activity and greatly increased HIS in serum, gut microbiota, and brain. Moreover, exogenous HIS demonstrated indirect neuroprotective effects through metabolizing to histamine. Notably, vagus nerve severance in I/R rats was performed to investigate HIS's neuroprotective mechanism. The results innovatively revealed that PNS could promote HIS synthesis in gut by enhancing L.Reu proportion, thereby increasing intracerebral HIS through peripheral pathway. Consequently, our data provided novel insights into HIS metabolism mediated by L.Reu in the pathogenesis and treatment of IS.

Objective To explore the effect of triglyceride glucose(TyG)index and uric acid level on carotid athero-sclerosis in patients with type 2 diabetes mellitus(T2DM),and build a risk nomogram model for T2DM complicated with carotid atherosclerosis.Methods A total of 125 T2DM patients admitted to the First People's Hospital of Xinxiang City from January 2020 to December 2022 were selected as the research subjects.The patients were divided into the carotid atherosclerosis group(n=33)and the non-carotid atherosclerosis group(n=92)according to whether they had carotid atherosclerosis.The general clinical data such as gender,age,course of disease,body mass index(BMI),diastolic blood pressure(DBP),and systolic blood pressure(SBP)of patients were collected through the electronic medical record system.The levels of triglycerides(TG),fasting plasma glucose(FPG),creatinine(Cr),fasting insulin(FINS),glycosylated hemoglobin(HbA1c),high-densi-ty lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),and total cholesterol(TC)were measured by using a fully automatic biochemical analyzer.The uric acid level was determined by using the uricase method.The risk factors of carotid atherosclerosis in T2DM patients were analyzed through univariate and multivariate logistic regression.R3.5.3 software was used to draw a nomogram model to predict carotid atherosclerosis in T2DM patients.The predictive performance of the nomogram model was validated by the receiver operating curve(ROC),and the accuracy of the model was tested by the Bootstrap method.Additionally,a total of 27 T2DM patients admitted to the First People's Hospital of Xinxiang City from January 2023 to June 2023 were selected to validate the predictive performance of the model.Results The univariate analysis showed that there were significant differences in the age,course of disease,Cr,FINS,FPG,TC,TG,TyG index and uric acid level of patients between the carotid atherosclerosis group and the non-carotid atherosclerosis group(P＜0.05);there was no statisti-cally significant difference in gender,BMI,DBP,SBP,HbA1c,HDL-C and LDL-C of patients between the two groups(P＞0.05).Multivariate logistic regression analysis showed that age ≥60 years,high Cr level,low FINS level,high TC level,high TyG index and high uric acid level were the risk factors for carotid atherosclerosis in T2DM patients(P＜0.05).The.The validation results of Bootstrap method for validating the early warning model established based on age,Cr,FINS,TC,TyG index and uric acid level to predict the risk of carotid atherosclerosis in T2DM patients showed that the C-index of the model was 0.762(95％confidence interval:0.728-0.808),and the area under the ROC curve,sensitivity,and specificity were 0.779,77.34％,and 82.46％,respectively.Among the 27 T2DM patients,8 cases developed carotid atherosclerosis,while the risk nomogram model predicted 7 cases of carotid arteriosclerosis.The ROC curve showed that the area under the curve,sensitivity and specificity were 0.785,80.47％and 75.36％,respectively.Conclusion High TyG index and high uric acid level are risk factors for T2DM patients with carotid atherosclerosis.The risk nomogram model constructed based on TyG index,uric acid level,age,Cr,FINS and TC has a good predictive effect on carotid atherosclerosis in T2DM patients.

Objective To understand the basic situation of blood transfusion adverse reactions to provide a basis for more ra-tionally and safely using blood in clinic.Methods A total of 24 409 cases of blood transfusion in this hospital during 2012?2015 were retrospectively analyzed.The report forms of blood transfusion adverse reactions served as the criterion for conducting the sta-tistics.Results There were 214 case of transfusion adverse reaction in this hospital during these 4 years.The incidence rate was 0. 88%.Among 5 blood components,the adverse reaction occurrence by frozen plasma was maximal,followed by single donor PLT and suspension red blood cells,which by the washed red blood cells was lowest.The blood adverse reactions showed the decreasing trend with the time and increasing trend with the age group increase.The transfusion reactions had no statistical difference between sexes and among blood types(P>0.05).Conclusion Clinical medical units should fully recognize the risk of blood transfusion,set up a standardized blood usage system and strictly implement,strengthen the blood transfusion knowledge training,advocate the au-tologous blood transfusion and clinically scientific,reasonable and safe blood usage.

Objective To investigate the effects of liraglutide on the proliferation and differentiation of mouse pre-osteoblasts MC3T3-E1 exposed to higher glucose concentration. Methods MC3T3-E1 cells were cultured and divided into control group and liraglutide group. In liraglutide group, cells were treated with different liraglutide concentrations (10-9 mol/L, 10-8 mol/L, and 10-7 mol/L, respectively) for 48 hours. Cell proliferation was tested with CCK-8. The mRNA expressions of typeⅠcollagen (COL-Ⅰ), osteopontin (OPN), and alkaline phosphase ( ALP) were detected using semi-quantitative RT-PCR. Results ( 1 ) Compared to control group, the proliferation rate of different liraglutide concentration groups (10-9 mol/L, 10-8 mol/L, and 10-7 mol/L) increased significantly (all P<0. 05), the proliferation rate was the highest in 10-8mol/L liraglutide group. (2)The expression of COL-Ⅰ, OPN, and ALP mRNA in liraglutide groups were higher than those in control group (all P<0. 05). The optimal concentration of liraglutide was 10-8 mol/L. Conclusion Liraglutide within a certain concentration range may improve the proliferation and differentiation of mouse pre-osteoblasts MC3T3-E1.

[Summary] To investigate the effect of gliclazide on the proliferation and differentiation in MC3T3-E1 cells exposed to normal glucose concentration by applying CCK-8 and RT-PCR. Gliclazide improved the proliferation and stimulated COL-I, OPN and Runx2 mRNA expression in MC3T3-E1 cells, the best concentration of gliclazide was 20μmol/ L, the expression of COL-1 and OPN mRNA had a significant positive correlation with Runx2 binding activity.

After MC3T3-E1 cells were treated with 1,10,and 20 μmol/L glibenclamide for 48 h,the proliferation rate of cells was detected by CCK-8 assay.Flow cytometry was used to test cell apoptosis.The mRNA expressions of collagen I (COL-1) and osteopontin (OPN) were tested by realtime fluorescence quantitative PCR.Western blot was used to detect the expression levels of apoptosis-related proteins Bax and Bcl-2.The results showed that compared with the control group,the proliferation rate of MC3T3-E1 cells was gradually increased (P＜0.05),the apoptosis rate decreased (P ＜ 0.05),the expressions of COL-1 mRNA,OPN mRNA,and Bcl-2 protein were progressively raised (P＜0.05),and the expression of Bax protein were gradually decreased (P＜0.05) along with increasing concentration of glyburide.It suggested that glibenclamide could promote the proliferation and differentiation of MC3T3-E1 cells in high glucose and may inhibit apoptosis in a concentration-dependent manner within a certain range.

Objective To investigate the effects of TLR7 on imiquimod induced apoptosis of THP-1 derived macrophages.Methods Three cell lines ( THP-1 derived macrophages, MDCK cell line and HUVEC cell line) with different capabilities of expressing TLR7 were selected.The survival rates of cells af-ter the treatment with different concentrations of imiquimod were detected by MTT assay.The levels of IL-6 in the supernatants of TLR7 inhibitor chloroquine or TLR7-siRNA treated cells were detected by enzyme-linked immunosorbent assay.The apoptosis of cells was detected by flow cytometry after inhibiting the ex-pression of TLR7.Results Imiquimod induced the apoptosis of THP-1 derived macrophages, MDCK cell lines and HUVEC cell lines.The levels of IL-6 were significantly decreased as the expression of TLR7 was inhibited by treating THP-1 derived macrophages with chloroquine or TLR7-siRNA.Treating THP-1 derived macrophages with chloroquine or TLR7-siRNA did not affect the cell apoptosis induced by imiquimod.Con-clusion Imiquimod could induce the apoptosis of THP-1 derived macrophages through TLR7 independent pathway.