Objective To explore the mechanism of the thioredoxin-interacting protein(TXNIP)/thioredoxin-1(Trx-1)pathway in regulating the polarization of retinal microglia in rats after retinal ischemia-reperfusion(RIR)in rats,and to provide new ideas for the prevention and treatment of retinal ischemia reperfusion injury(RIRI).Methods For-ty-two healthy adult male Sprague-Dawley rats were randomly divided into a Sham group,a RIRI group and a TXNIP siRNA group.The right eye of the rats was experimented.For RIRI and TXNIP siRNA groups,RIRI models were established using the anterior chamber high intraocular pressure method.Rats in the TXNIP siRNA group were given the intravitreal injection of TXNIP siRNA 3 d before modeling.Hematoxylin-eosin(HE)staining was used to analyze retinal histopathologic changes of rats in all groups 24 h after modeling.Immunohistochemical staining of brain-specific homeobox/POU domain proteins 3A(Brn-3a)was made to count the number of retinal ganglion cells(RGCs).The dynamical changes in the number of TXNIP+cells 6 h,24 h,72 h and 7 d after modelling were analyzed through immunohistochemical staining in the RIRI group.The retinal microglia polarization and changes in the expression of TXNIP and Trx-1 proteins in each group were de-tected by double immunofluorescence staining and Western blot 24 h after modeling.Results HE staining results showed that 24 h after modelling,the retinal cells were disordered and the inner retinal layer was thickened and swelled in RIRI and TXNIP siRNA groups,compared with those in the Sham group(all P＜0.05).Immunohistochemical staining results of Brn-3a showed that 24 h after modeling,the number of Brn-3a+cells in RIRI and TXNIP siRNA groups significantly decreased,compared with that in the Sham group(both P＜0.05).The number of Brn-3a+cells in the TXNIP siRNA group was signifi-cantly higher than that in the RIRI group(P＜0.05).Immunohistochemical staining results of TXNIP at different time points after modeling showed that the expression of TXNIP+proteins started to increase 6 h after modeling.The TXNIP+protein level reached a peak at 24 h and then decreased gradually.Western blot results revealed that 24 h after modeling,RIRI and TXNIP siRNA groups had significantly higher TXNIP levels and significantly lower Trx-1 levels than the Sham group(all P＜0.05).Compared with those in the RIRI group,the expression of TXNIP proteins was significantly lower and the expression of Trx-1 proteins was significantly higher in the TXNIP siRNA group(both P＜0.05).Double immunofluores-cence staining showed that 24 h after modeling,Iba1+/CD206+cells were significantly more and Iba1+/CD16+cells were significantly less in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).RIRI and TXNIP siRNA groups had significantly more Ibal+/TXNIP+cells and significantly less Iba1+/Trx-1+cells than the Sham group(both P＜0.05).The number of Iba1+/TXNIP+cells was significantly lower and the number of Iba1+/Trx-1+cells was significantly higher in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).Conclusion RIR activates the TXNIP/Trx-1 path-way to induce the activation of retinal microglia and regulate the polarization of microglia,thereby resulting in RIRI in rats.

Objective:To analyze the contamination characteristics and pathogenic potential of Listeria monocytogenes (LM) in pre-packaged and refrigerated ready-to-eat cooked meat products in Chengdu City. Methods:From September 2022 to May 2023, pre-packaged and refrigerated ready-to-eat cooked meat products were collected from six districts and counties in Chengdu City. Qualitative and quantitative determination of LM was performed on these samples. Whole genome sequencing was carried out on the isolated strains. Different ST strains were selected for cell adhesion and invasion experiments. The results were expressed as adhesion rate and invasion rate. One-way ANOVA was used for comparison between groups, and the Dunnett t-test was used for pairwise comparison. Results:A total of 145 samples were collected, and LM was detected and isolated in 29 samples. The total detection rate was 20.00%, and the detection rate of braised pork was the highest (68.18%). The contamination level of LM in 9 samples (31.03%) was greater than 100.00 MPN/g. The 29 strains of Listeria monocytogenes belonged to 7 STs, including ST 3 (27.59%), ST 8 (17.24%) and ST 87 (13.79%). The strain of ST 87 carried Listeria pathogenicity islands 4 (LIPI-4), which was a highly virulent strain. The medium and high virulence strains accounted for 79.31%. Some moderately virulent and highly virulent strains of ST 8 and ST 87 were closely related to clinical patient strains. Some LM isolates of the same ST type had little SNP differences (1-6) in the same manufacturer at different stages. In vitro cell experiments showed that the highly virulent strain ST 87 had the strongest adhesion and invasion ability towards Caco-2 cells. Conclusion:The pre-packaged and refrigerated cooked meat products have a high contamination rate. Some samples have high contamination levels and carry medium and high virulence strains, posing potential health risks to human beings. LM residues continue to persist in some manufacturers′ production and processing stages.

Objective To constructed a predictive model of gastric adenocarcinoma prognosis-related proteins by using TCPA and UCSC Xena databases to obtain sample information.And its application value was tested in order to provide reference for clinical treatment and related mechanism research.Methods Using R project,univariate Cox analysis and LASSO-Cox regression analysis were used to select the proteins related to the prognosis of gastric adenocarcinoma,and the risk scores were calculated.According to the risk scores,the patients were divided into high risk group and low risk group for survival analysis.At the same time,draw the ROC curve of the risk score and the protein heat map to evaluate the predictive ability of these proteins.Then,risk scores were combined with other clinical phenotype data of the patients,in order to establish a prognostic model through multivariate regression analysis,and draw a nomogram.The accuracy of the prognostic model was evaluated by C-index,ROC curve and decision curve.GO enrichment analysis and KEGG enrichment analysis were carried out to explore the proteins involved in biological processes.Results Fifteen differential proteins(CKIT,CHK1,CLAUDIN7,COLLAGENVI,DVL3,EGFR_pY1173,ERALPHA_pS11,NFKBP65_pS536,SYK,ETS1,MYOSINIIA_pS1943,P21,P90RSK,RAPTOR,XBP1)were selected by univariate Cox analysis and LASSO-Cox analysis,for calculating the protein risk scores.Through multivariate regression analysis,five factors including age,gender,M stage,N stage and risk score were selected to construct a prognostic model.The results of survival analysis showed that there were significant differences in overall survival between high and low risk groups(P＜0.0001).The nomogram can be used to predict 3-year and 5-year survival rate of patients,and it is considered to have good predictive value(C-index=0.7257).The ROC curve showed that the model exhibited stable sensitivity and specificity in prediction(AUC=0.79 and 0.88).Conclusion The prognostic model of gastric adenocarcinoma prognosis-related proteins can be used to predict the clinical prognosis of patients,and the research results will help to further guide clinical treatment,which could provide a reference for personalized treatment and management.

Objective:To analyze the contamination characteristics and pathogenic potential of Listeria monocytogenes (LM) in pre-packaged and refrigerated ready-to-eat cooked meat products in Chengdu City. Methods:From September 2022 to May 2023, pre-packaged and refrigerated ready-to-eat cooked meat products were collected from six districts and counties in Chengdu City. Qualitative and quantitative determination of LM was performed on these samples. Whole genome sequencing was carried out on the isolated strains. Different ST strains were selected for cell adhesion and invasion experiments. The results were expressed as adhesion rate and invasion rate. One-way ANOVA was used for comparison between groups, and the Dunnett t-test was used for pairwise comparison. Results:A total of 145 samples were collected, and LM was detected and isolated in 29 samples. The total detection rate was 20.00%, and the detection rate of braised pork was the highest (68.18%). The contamination level of LM in 9 samples (31.03%) was greater than 100.00 MPN/g. The 29 strains of Listeria monocytogenes belonged to 7 STs, including ST 3 (27.59%), ST 8 (17.24%) and ST 87 (13.79%). The strain of ST 87 carried Listeria pathogenicity islands 4 (LIPI-4), which was a highly virulent strain. The medium and high virulence strains accounted for 79.31%. Some moderately virulent and highly virulent strains of ST 8 and ST 87 were closely related to clinical patient strains. Some LM isolates of the same ST type had little SNP differences (1-6) in the same manufacturer at different stages. In vitro cell experiments showed that the highly virulent strain ST 87 had the strongest adhesion and invasion ability towards Caco-2 cells. Conclusion:The pre-packaged and refrigerated cooked meat products have a high contamination rate. Some samples have high contamination levels and carry medium and high virulence strains, posing potential health risks to human beings. LM residues continue to persist in some manufacturers′ production and processing stages.

Objective To constructed a predictive model of gastric adenocarcinoma prognosis-related proteins by using TCPA and UCSC Xena databases to obtain sample information.And its application value was tested in order to provide reference for clinical treatment and related mechanism research.Methods Using R project,univariate Cox analysis and LASSO-Cox regression analysis were used to select the proteins related to the prognosis of gastric adenocarcinoma,and the risk scores were calculated.According to the risk scores,the patients were divided into high risk group and low risk group for survival analysis.At the same time,draw the ROC curve of the risk score and the protein heat map to evaluate the predictive ability of these proteins.Then,risk scores were combined with other clinical phenotype data of the patients,in order to establish a prognostic model through multivariate regression analysis,and draw a nomogram.The accuracy of the prognostic model was evaluated by C-index,ROC curve and decision curve.GO enrichment analysis and KEGG enrichment analysis were carried out to explore the proteins involved in biological processes.Results Fifteen differential proteins(CKIT,CHK1,CLAUDIN7,COLLAGENVI,DVL3,EGFR_pY1173,ERALPHA_pS11,NFKBP65_pS536,SYK,ETS1,MYOSINIIA_pS1943,P21,P90RSK,RAPTOR,XBP1)were selected by univariate Cox analysis and LASSO-Cox analysis,for calculating the protein risk scores.Through multivariate regression analysis,five factors including age,gender,M stage,N stage and risk score were selected to construct a prognostic model.The results of survival analysis showed that there were significant differences in overall survival between high and low risk groups(P＜0.0001).The nomogram can be used to predict 3-year and 5-year survival rate of patients,and it is considered to have good predictive value(C-index=0.7257).The ROC curve showed that the model exhibited stable sensitivity and specificity in prediction(AUC=0.79 and 0.88).Conclusion The prognostic model of gastric adenocarcinoma prognosis-related proteins can be used to predict the clinical prognosis of patients,and the research results will help to further guide clinical treatment,which could provide a reference for personalized treatment and management.

Objective To explore the mechanism of the thioredoxin-interacting protein(TXNIP)/thioredoxin-1(Trx-1)pathway in regulating the polarization of retinal microglia in rats after retinal ischemia-reperfusion(RIR)in rats,and to provide new ideas for the prevention and treatment of retinal ischemia reperfusion injury(RIRI).Methods For-ty-two healthy adult male Sprague-Dawley rats were randomly divided into a Sham group,a RIRI group and a TXNIP siRNA group.The right eye of the rats was experimented.For RIRI and TXNIP siRNA groups,RIRI models were established using the anterior chamber high intraocular pressure method.Rats in the TXNIP siRNA group were given the intravitreal injection of TXNIP siRNA 3 d before modeling.Hematoxylin-eosin(HE)staining was used to analyze retinal histopathologic changes of rats in all groups 24 h after modeling.Immunohistochemical staining of brain-specific homeobox/POU domain proteins 3A(Brn-3a)was made to count the number of retinal ganglion cells(RGCs).The dynamical changes in the number of TXNIP+cells 6 h,24 h,72 h and 7 d after modelling were analyzed through immunohistochemical staining in the RIRI group.The retinal microglia polarization and changes in the expression of TXNIP and Trx-1 proteins in each group were de-tected by double immunofluorescence staining and Western blot 24 h after modeling.Results HE staining results showed that 24 h after modelling,the retinal cells were disordered and the inner retinal layer was thickened and swelled in RIRI and TXNIP siRNA groups,compared with those in the Sham group(all P＜0.05).Immunohistochemical staining results of Brn-3a showed that 24 h after modeling,the number of Brn-3a+cells in RIRI and TXNIP siRNA groups significantly decreased,compared with that in the Sham group(both P＜0.05).The number of Brn-3a+cells in the TXNIP siRNA group was signifi-cantly higher than that in the RIRI group(P＜0.05).Immunohistochemical staining results of TXNIP at different time points after modeling showed that the expression of TXNIP+proteins started to increase 6 h after modeling.The TXNIP+protein level reached a peak at 24 h and then decreased gradually.Western blot results revealed that 24 h after modeling,RIRI and TXNIP siRNA groups had significantly higher TXNIP levels and significantly lower Trx-1 levels than the Sham group(all P＜0.05).Compared with those in the RIRI group,the expression of TXNIP proteins was significantly lower and the expression of Trx-1 proteins was significantly higher in the TXNIP siRNA group(both P＜0.05).Double immunofluores-cence staining showed that 24 h after modeling,Iba1+/CD206+cells were significantly more and Iba1+/CD16+cells were significantly less in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).RIRI and TXNIP siRNA groups had significantly more Ibal+/TXNIP+cells and significantly less Iba1+/Trx-1+cells than the Sham group(both P＜0.05).The number of Iba1+/TXNIP+cells was significantly lower and the number of Iba1+/Trx-1+cells was significantly higher in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).Conclusion RIR activates the TXNIP/Trx-1 path-way to induce the activation of retinal microglia and regulate the polarization of microglia,thereby resulting in RIRI in rats.

In the past decades, significant progress has been achived in cartilage regeneration. The traditional techniques for constructing tissue engineering cartilage scaffold mainly include pore agent method (or template method), phase separation method, gas foaming method, freeze-drying method, electrospinning method, etc. Cartilage is heterogeneous, and it is difficult for traditional scaffolds to simulate the high anisotropy of cartilage. Therefore, functional regeneration of cartilage is challenging. With the progress of three-dimensional(3D) printing technology, it is possible to prepare functional bionic scaffolds with fine structure and gradient changes through co-deposition of biomaterials, cells and active biomolecules, so as to achieve functional cartilage regeneration. This article reviewed 3D printing technology of cartilage tissue engineering, and the application of 3D printing technology in cartilage regeneration at different anatomical positions (articular cartilage, auricle cartilage, nasal cartilage). In addition, the importance of preparing bionic constructs with regional structure gradient and regional composition gradient was discussed. 3D bioprinting technology, 4D printing techniques, smart biomaterials brought hope for the construction of bionic tissues and organs.

In order to promote the innovative application of Sanjiao theory and Yingwei theory， this paper tries to apply the ''Sanjiao-Yingwei'' Qi transformation theory to the treatment of tumor diseases， integrating it with T cell exhaustion mechanism to elaborate on its scientific connotation and using network pharmacology and bioinformatics to elucidate the correlation between the anti-tumor mechanism of ''Sanjiao-Yingwei'' Qi transformation and T cell exhaustion. The ''Sanjiao-Yingwei'' Qi transformation function is closely related to the immunometabolic ability of the human body， and the ''Sanjiao-Yingwei'' Qi transformation system constitutes the immunometabolic exchange system within and outside the cellular environment. Cancer toxicity is generated by the fuzzy Sanjiao Qi， and the long-term fuzzy Sanjiao Qi is the primary factor leading to T cell exhaustion， which is related to the long-term activation of T cell receptors by the high tumor antigen load in the tumor microenvironment. Qi transformation malfunction of the Sanjiao produces phlegm and collects stasis， which contributes to T cell exhaustion and is correlated with nutrient deprivation， lipid accumulation， and high lactate levels in the immunosuppressed tumor microenvironment， as well as with the release of transforming growth factor-β and upregulated expression of programmed death receptor-1 by tumor-associated fibroblasts and platelets in the tumor microenvironment. Ying and Wei damage due to Sanjiao Qi transformation malfunction is similar to the abnormal manifestations such as progressive loss of exhausted T cell effector function and disturbance of cellular energy metabolism. Guizhi decoction， Shengming decoction， and Wendan decoction can correct T cell exhaustion and exert anti-tumor effects through multi-target and multi-pathways by regulating ''Sanjiao-Yingwei'' Qi transformation， and hypoxia inducible factor-1α （HIF-1α） may be one of the main pathways to correct T cell exhaustion. It was found that HIF-1α may be one of the important prognostic indicators in common tumors by bioinformatics. The use of the ''Sanjiao-Yingwei'' Qi transformation method may play an important part in improving the prognosis of tumor patients in clinical practice.

BACKGROUND:Exercise has a regulatory effect on intestinal flora dysbiosis,which can effectively protect the beneficial flora and improve the intestinal environment.However,the effect of treadmill exercise on the structure and diversity of intestinal microbial community in Parkinson's disease and the specific mechanism are not clear. OBJECTIVE:Using 16S rDNA technique to analyze the effect of treadmill exercise on the structure and diversity of the intestinal flora of rats with Parkinson's disease,and to investigate the mechanism of non-pharmacological treadmill exercise to improve Parkinson's disease. METHODS:Twelve of the 18 Sprague-Dawley rats were randomly selected to make animal models of Parkinson's disease using unilateral 2-point nigrostriatal injection of 6-hydroxydopamine.The remaining six rats were used as sham-operation group,which were injected with the same dose of saline containing 0.2%ascorbic acid using the same positioning and injection method.After successful modeling,12 rats with Parkinson's disease were randomly divided into model group and treadmill exercise group(n=6 per group).The treadmill exercise group was subjected to a middle and low intensity tread mill exercise,10 m/min,30 minutes per day,5 days per week for 4 weeks.Fresh feces were collected and stored in liquid nitrogen 24 hour after the last exercise session,and the changes in fecal flora were analyzed by 16S rDNA sequencing technique. RESULTS AND CONCLUSION:Treadmill exercise significantly improved behavior and nigrostriatal tyrosine hydroxylase-positive cell expression in rats with Parkinson's disease model and alleviated changes in the structure and diversity of the gut microbial community caused by Parkinson's disease,increased the number of operational taxonomic units and modulated Alpha and Beta diversity in rats.At the phylum and genus levels,the abundance ratio of Firmicutes/Bacteroidetes in the model group decreased compared with the sham-operated group,while beneficial bacteria such as Prevotella,Bacteroides,and Clostridium_XlV increased significantly after treadmill exercise.To conclude,treadmill exercise has a significant modulating effect on behavioral abnormalities,toxic damage to dopaminergic neurons and gut microbial imbalance caused by Parkinson's disease,alleviates the symptoms of flora-related diseases,and has a positive effect on the improvement of Parkinson's disease.

In the past decades, significant progress has been achived in cartilage regeneration. The traditional techniques for constructing tissue engineering cartilage scaffold mainly include pore agent method (or template method), phase separation method, gas foaming method, freeze-drying method, electrospinning method, etc. Cartilage is heterogeneous, and it is difficult for traditional scaffolds to simulate the high anisotropy of cartilage. Therefore, functional regeneration of cartilage is challenging. With the progress of three-dimensional(3D) printing technology, it is possible to prepare functional bionic scaffolds with fine structure and gradient changes through co-deposition of biomaterials, cells and active biomolecules, so as to achieve functional cartilage regeneration. This article reviewed 3D printing technology of cartilage tissue engineering, and the application of 3D printing technology in cartilage regeneration at different anatomical positions (articular cartilage, auricle cartilage, nasal cartilage). In addition, the importance of preparing bionic constructs with regional structure gradient and regional composition gradient was discussed. 3D bioprinting technology, 4D printing techniques, smart biomaterials brought hope for the construction of bionic tissues and organs.