The objective of this study was to evaluate the protective effect and possible related mechanisms of Sophora subprostrate polysaccharide(SSP)on intestinal epithelial cell injury in-duced by Tert-Butyl hydroperoxide(TBHP).The optimal dose of TBHP and the safe concentra-tion range of SSP were determined using the MTT method.In this study,IPEC-J2 cells were divid-ed into five groups:the control group,the model group,the SSPL group,the SSPM group and the SSPH group,and the cell morphology,cell survival rate and LDH release rate were observed and measured.The content of intracellular reactive ROS was observed and determined by DCFH-DA staining.The content of MDA in the supernatant and the antioxidant index of cells were determined by the reagent kit.Transcriptome technology was employed to analyze the potential mechanisms by which SSP mitigates oxidative damage in IPEC-J2 cells.The results showed that treatment with 625 μmol/L TBHP for 2 h significantly reduced the activity of IPEC-J2 cells,markedly increased LDH release(P＜0.05),inhibited CAT superoxide SOD and glutathione GPX activities(P＜0.05),and significantly elevated MDA and ROS levels(P＜0.05).Compared to the model group,after SSP treatment,intracellular ROS levels were significantly reduced(P＜0.05),while CAT,SOD,and GPX activities were significantly increased(P＜0.05),and MDA content and LDH re-lease were significantly decreased(P＜0.05)in a dose-dependent manner.Transcriptome analysis revealed that TBHP treatment significantly altered the transcriptional profiles of IPEC-J2 cells,while SSP treatment could restore the transcriptional profiles of the damaged cells to a certain ex-tent.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)indicated that the differentially expressed genes between the CC and TBHP groups were significantly enriched in oxidative phosphorylation,ribosome,and other pathways.Meanwhile,the differentially expressed genes between the SSP and TBHP groups were mainly enriched in oxidative phosphorylation,ap-optosis,glyoxylate and dicarboxylate metabolism,and other pathways.These results suggest that TBHP may disrupt normal oxidative respiration in IPEC-J2 cells by affecting oxidative phospho-rylation and interfering with metabolism pathways involving glycine,serine,and threonine,leading to oxidative damage in intestinal epithelial cells.Conversely,SSP treatment may potentially restore oxidative phosphorylation processes,alleviate lysosomal damage,reduce cell apoptosis,and miti-gate oxidative damage in intestinal epithelial cells through modulation of oxidative phosphoryla-tion,apoptosis,and lysosomal pathways.This discovery provides a theoretical basis for the clinical application of SSP in alleviating oxidative damage in the porcine intestinal tract.

Foot-and-mouth disease (FMD) is one of the major animal infectious diseases in the world. All cloven-hoofed animals are susceptible to FMD. Vaccination is still the first choice for the prevention and control of FMD. mRNA vaccines can be rapidly designed, synthesized, and produced on a large scale in vitro, and they can induce effective protective immune responses, demonstrating the advantages of rapid development, easy preparation, and low biosafety risks. The design of untranslated regions is a key to enhancing the expression and efficacy of mRNA vaccines. In order to generate an efficient FMD mRNA vaccine, we designed three FMD P12A3C expression vectors with different untranslated regions and synthesized corresponding mRNAs. By comparing expression efficiency of these vectors and their mRNAs at different time points and in different cell lines, we found that the mRNA P12A3C-UTR3 had the best expression and universality. This study laid a foundation for the development of mRNA vaccines against FMD and provided a theoretical basis for the optimal sequence design of efficient mRNA.
Foot-and-Mouth Disease Virus/genetics* ; Animals ; RNA, Messenger/biosynthesis* ; Foot-and-Mouth Disease/immunology* ; Antigens, Viral/biosynthesis* ; Viral Vaccines/biosynthesis* ; Genetic Vectors/genetics* ; Cell Line ; Vaccines, DNA/immunology*

The objective of this study was to evaluate the protective effect and possible related mechanisms of Sophora subprostrate polysaccharide(SSP)on intestinal epithelial cell injury in-duced by Tert-Butyl hydroperoxide(TBHP).The optimal dose of TBHP and the safe concentra-tion range of SSP were determined using the MTT method.In this study,IPEC-J2 cells were divid-ed into five groups:the control group,the model group,the SSPL group,the SSPM group and the SSPH group,and the cell morphology,cell survival rate and LDH release rate were observed and measured.The content of intracellular reactive ROS was observed and determined by DCFH-DA staining.The content of MDA in the supernatant and the antioxidant index of cells were determined by the reagent kit.Transcriptome technology was employed to analyze the potential mechanisms by which SSP mitigates oxidative damage in IPEC-J2 cells.The results showed that treatment with 625 μmol/L TBHP for 2 h significantly reduced the activity of IPEC-J2 cells,markedly increased LDH release(P＜0.05),inhibited CAT superoxide SOD and glutathione GPX activities(P＜0.05),and significantly elevated MDA and ROS levels(P＜0.05).Compared to the model group,after SSP treatment,intracellular ROS levels were significantly reduced(P＜0.05),while CAT,SOD,and GPX activities were significantly increased(P＜0.05),and MDA content and LDH re-lease were significantly decreased(P＜0.05)in a dose-dependent manner.Transcriptome analysis revealed that TBHP treatment significantly altered the transcriptional profiles of IPEC-J2 cells,while SSP treatment could restore the transcriptional profiles of the damaged cells to a certain ex-tent.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)indicated that the differentially expressed genes between the CC and TBHP groups were significantly enriched in oxidative phosphorylation,ribosome,and other pathways.Meanwhile,the differentially expressed genes between the SSP and TBHP groups were mainly enriched in oxidative phosphorylation,ap-optosis,glyoxylate and dicarboxylate metabolism,and other pathways.These results suggest that TBHP may disrupt normal oxidative respiration in IPEC-J2 cells by affecting oxidative phospho-rylation and interfering with metabolism pathways involving glycine,serine,and threonine,leading to oxidative damage in intestinal epithelial cells.Conversely,SSP treatment may potentially restore oxidative phosphorylation processes,alleviate lysosomal damage,reduce cell apoptosis,and miti-gate oxidative damage in intestinal epithelial cells through modulation of oxidative phosphoryla-tion,apoptosis,and lysosomal pathways.This discovery provides a theoretical basis for the clinical application of SSP in alleviating oxidative damage in the porcine intestinal tract.

To investigate the alleviating effect of glycyrrhizic acid(GA)on chronic toxicity of afla-toxin B1(AFB1)in ducklings.Eighty 4-day-old ducklings were randomly divided into 4 groups,with 20 ducklings in each group,including a blank group,a model group(100 μg/kg AFB1),a high-concentration GA group(100 μg/kg AFB1+100 mg/kg GA),and a low-concentration GA group(100 μg/kg AFB1+50 mg/kg GA),with a trial period of 18 days.At the end of the experi-ment,the body weight of the ducklings in each group,AFB1-DNA content in the liver and liver tis-sues index,the biochemical indicators of liver function,and the pathological changes in liver tissues were examined.Additionally,the oxidative damage status of the liver tissues was eval-uated,and the mRNA expression levels of mitochondria-related apoptosis genes was detected.Com-pared with the control group,the body weight of ducklings in the model group decreased signifi-cantly(P＜0.01),AFB1-DNA content in the tissues increased significantly(P＜0.05),liver swell-ing and yellowing were observed,the liver index increased significantly(P＜0.01),as did the levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)(P＜0.01).The AST/ALT levels also increased significantly(P＜0.01),while the serum total protein(TP)content de-creased significantly(P＜0.01).The liver tissues showed a large amount of inflammatory cell infil-tration,vacuolar degeneration,fibrotic changes and apoptosis.The activities of catalase(CAT),glutathione S-transferase(GST),the total antioxidant capacity(T-AOC)of the duckling liver all decreased significantly(P＜0.01)and the mRNA expression of mitochondria-related apoptosis genes Bax,Cyt-c,Caspase-3,and Caspase-9 significantly increased(P＜0.01).Compared with the model group,GA treatment significantly increased the body weight of the model ducklings(P＜0.01),reduced AFB1-DNA content in the tissue,significantly reduced their liver index(P＜0.05),visibly restored the liver's apparent status,effectively reversed the abnormal changes in serum liver function indicators,improved the pathological changes in liver tissue histology,enhanced liver an-tioxidant function,significantly decreased expression of Bax,Cyt-c,Caspase-3,and Caspase-9 mR-NA(P＜0.05).Further correlation analysis showed that mRNA expression levels of Bax,Cyt-c,Caspase-3,and Caspase-9 in duck liver tissues were positively correlated with liver index,AFB1-DNA content,AST content,ALT content,AST/ALT ratio,and MDA content,and negatively cor-related with body weight,TP content,SOD activity,CAT activity,GST activity,and T-AOC activi-ty in ducklings.In conclusion,GA may alleviate liver damage to relieve duckling AFB1 chronic tox-icity by inhibiting the mitochondrial pathway of apoptosis.

Background/Aims: There may be many predictors of anticoagulation-related gastrointestinal bleeding (GIB), but until now, systematic reviews and assessments of the certainty of the evidence have not been published. We conducted a systematic review to identify all risk factors for anticoagulant-associated GIB to inform risk prediction in the management of anticoagulation- related GIB. Methods: A systematic review and meta-analysis were conducted to search PubMed, EMBASE, Web of Science, and Cochrane Library databases (from inception through January 21, 2022) using the following search terms: anticoagulants, heparin, warfarin, dabigatran, rivaroxaban, apixaban, DOACs, gastrointestinal hemorrhage, risk factors. According to inclusion and exclusion criteria, studies of risk factors for anticoagulation-related GIB were identified. Risk factors for anticoagulant-associated GIB were used as the outcome index of this review. Results: We included 34 studies in our analysis. For anticoagulant-associated GIB, moderate-certainty evidence showed a probable association with older age, kidney disease, concomitant use of aspirin, concomitant use of the antiplatelet agent, heart failure, myocardial infarction, hematochezia, renal failure, coronary artery disease, helicobacter pylori infection, social risk factors, alcohol use, smoking, anemia, history of sleep apnea, chronic obstructive pulmonary disease, international normalized ratio (INR), obesity et al. Some of these factors are not included in current GIB risk prediction models. such as anemia, co-administration of gemfibrozil, co-administration of verapamil or diltiazem, INR, heart failure, myocardial infarction, etc. Conclusions The study found that anemia, co-administration of gemfibrozil, co-administration of verapamil or diltiazem, INR, heart failure, myocardial infarction et al. were associated with anticoagulation-related GIB, and these factors were not in the existing prediction models. This study informs risk prediction for anticoagulant-associated GIB, it also informs guidelines for GIB prevention and future research.

Objective:To investigate the relationship between serum osteopontin (OPN), bone morphogenetic protein 2 (BMP2), retinol binding protein 4 (RBP4) and renal function and bone mineral density in patients with diabetes nephropathy (DN).Methods:A total of 120 patients with DN diagnosed in the Affiliated Hospital of Jining Medical University from January 2020 to December 2021 were selected as the DN group, 60 patients with simple diabetes as the type 2 diabetes mellitus (T2DM) group, and 60 subjects with normal glucose tolerance test as the control group. The serum OPN, BMP2, RBP4, low bone mineral density (LBMD), femoral neck bone density (FNBMD) and renal function indicators of the three groups were compared. According to the urinary albumin excretion rate (UAER) of DN patients, the patients were divided into microalbuminuria DN group (71 cases) and massive albuminuria DN group (49 cases), and stratified comparison was made. The simple linear correlation analysis was used to analyze the OPN of DN patients. BMP2, RBP4, renal function and bone mineral density.Results:The fasting blood glucose (FPG), glycated hemoglobin (HbA 1c), serum creatinine (Scr), UAER, and cystatin (CysC) levels of DN group patients were significantly higher than those of T2DM group and control group, and the differences were statistically significant (all P<0.05); The FPG and HbA 1c in the T2DM group were higher than those in the control group, and the differences were statistically significant (all P<0.05); The OPN and BMP2 of DN group patients were higher than those of T2DM group and control group, while the RBP4, LBMD, FNBMD of DN group were lower than those of T2DM group and control group, and the differences were statistically significant (all P<0.05); The OPN and BMP2 of the T2DM group were higher than those of the control group, while RBP4 was lower than that of the control group, and the differences were statistically significant (all P<0.05); The levels of FPG, HbA 1c, Scr, UAER, and CysC in patients with macroalbuminuria DN were significantly higher than those in patients with microalbuminuria DN, and the differences were statistically significant (all P<0.05); The OPN and BMP2 of patients in the large albuminuria DN group were higher than those in the microalbuminuria DN group, while the RBP4, LBMD, and FNBMD of patients in the large albuminuria DN group were lower than those in the microalbuminuria DN group, and the differences were statistically significant (all P<0.05). The OPN of DN group patients was positively correlated with Scr, UAER, and CysC (all P<0.05), while BMP2 was positively correlated with UAER and CysC (all P<0.05); The OPN and BMP2 of DN group patients were negatively correlated with LBMD and FNBMD (all P<0.05), while RBP4 was positively correlated with LBMD and FNBMD (all P<0.05). Conclusions:OPN, BMP2, RBP4 are closely related to the degree of renal function impairment and bone loss in DN patients, and can to some extent reflect the degree of bone metabolism and osteoporosis in T2DM patients.

Lipid nanoparticles serve as a promising drug delivery system due to the good biocompatibility, non-immunogenicity, and high drug loading efficiency. However, unmodified lipid nanoparticles have limitations such as poor stability, easy hydrolysis, and rapid removal. To overcome these shortcomings, researchers have developed peptide modification, antibody modification, ligand modification, nucleic acid aptamer modification, and polysaccharide modification for lipid nanoparticles. Polysaccharides are a class of natural polymers, and the polysaccharide-modified lipid nanoparticles exhibit good biocompatibility, precise targeting, and low toxicity. Therefore, polysaccharide-modified lipid nanoparticles demonstrate great potential in clinical treatment. This review summarizes the preparation and application of polysaccharide-modified lipid nanoparticles, aiming to provide a reference for further research and development of new lipid nanoparticles.
Polysaccharides/chemistry* ; Nanoparticles/chemistry* ; Lipids/chemistry* ; Drug Delivery Systems ; Humans ; Drug Carriers/chemistry*

To investigate the alleviating effect of glycyrrhizic acid(GA)on chronic toxicity of afla-toxin B1(AFB1)in ducklings.Eighty 4-day-old ducklings were randomly divided into 4 groups,with 20 ducklings in each group,including a blank group,a model group(100 μg/kg AFB1),a high-concentration GA group(100 μg/kg AFB1+100 mg/kg GA),and a low-concentration GA group(100 μg/kg AFB1+50 mg/kg GA),with a trial period of 18 days.At the end of the experi-ment,the body weight of the ducklings in each group,AFB1-DNA content in the liver and liver tis-sues index,the biochemical indicators of liver function,and the pathological changes in liver tissues were examined.Additionally,the oxidative damage status of the liver tissues was eval-uated,and the mRNA expression levels of mitochondria-related apoptosis genes was detected.Com-pared with the control group,the body weight of ducklings in the model group decreased signifi-cantly(P＜0.01),AFB1-DNA content in the tissues increased significantly(P＜0.05),liver swell-ing and yellowing were observed,the liver index increased significantly(P＜0.01),as did the levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)(P＜0.01).The AST/ALT levels also increased significantly(P＜0.01),while the serum total protein(TP)content de-creased significantly(P＜0.01).The liver tissues showed a large amount of inflammatory cell infil-tration,vacuolar degeneration,fibrotic changes and apoptosis.The activities of catalase(CAT),glutathione S-transferase(GST),the total antioxidant capacity(T-AOC)of the duckling liver all decreased significantly(P＜0.01)and the mRNA expression of mitochondria-related apoptosis genes Bax,Cyt-c,Caspase-3,and Caspase-9 significantly increased(P＜0.01).Compared with the model group,GA treatment significantly increased the body weight of the model ducklings(P＜0.01),reduced AFB1-DNA content in the tissue,significantly reduced their liver index(P＜0.05),visibly restored the liver's apparent status,effectively reversed the abnormal changes in serum liver function indicators,improved the pathological changes in liver tissue histology,enhanced liver an-tioxidant function,significantly decreased expression of Bax,Cyt-c,Caspase-3,and Caspase-9 mR-NA(P＜0.05).Further correlation analysis showed that mRNA expression levels of Bax,Cyt-c,Caspase-3,and Caspase-9 in duck liver tissues were positively correlated with liver index,AFB1-DNA content,AST content,ALT content,AST/ALT ratio,and MDA content,and negatively cor-related with body weight,TP content,SOD activity,CAT activity,GST activity,and T-AOC activi-ty in ducklings.In conclusion,GA may alleviate liver damage to relieve duckling AFB1 chronic tox-icity by inhibiting the mitochondrial pathway of apoptosis.

Objective:To investigate the current status of polypharmacy among elderly outpatients with 4 types of chronic diseases such as hypertension, diabetes mellitus, coronary atherosclerotic heart disease, and cerebrovascular disease.Methods:A retrospective study was conducted on the drug use of elderly (≥65 years old) outpatients with hypertension, diabetes mellitus, coronary atherosclerotic heart disease, and cerebrovascular disease with data of Beijing Municipal Health Insurance Centre database from July 2017 to September 2017. The included patients had at least 1 of 4 types of chronic diseases. Polypharmacy was defined as ≥5 different types of medication at the first visit, and non-polypharmacy was defined as <5 types of medication. The number and severity of comorbidity were quantified using the Charlson Comorbidity Index (CCI), and the prognosis of patient was evaluated at 4 levels of 0, 1, 2, and ≥3 scores. The larger the value, the more severe the disease. Based on the Beers Criteria 2015, the potential inappropriate medication (PIM) was identified using the prescription review system of Puhua Health.Results:A total of 405 608 patients were included in this study, with a median age of 74 (65-107) years, and 204 219 patients (50.35%) were female. According to the type of medication used by patients, they were divided into polypharmacy group (113 594 cases, 28.01%) and non-polypharmacy group (292 014 cases, 71.99%). The CCI of the polypharmacy group was significantly higher than that of the non-polypharmacy group ( P<0.001). The proportion of patients with 0, 1, 2, and ≥3 scores in the polypharmacy group was significantly higher than that of the non-polypharmacy group, and the differences were statistically significant (all P<0.001). In terms of comorbidity, the proportions of patients among the 4 types of chronic diseases were higher in the polypharmacy group than in the non-polypharmacy group ( P<0.001). In terms of concomitant diseases, the proportion of patients with hyperlipidemia, cognitive impairment, heart failure, and osteoporosis in the polypharmacy group was higher than that in the non-polypharmacy group (all P<0.001). In terms of medical treatment behaviour, the median number of medical visits was higher in the polypharmacy group than in the non-polypharmacy group [2(1,3) vs. 1(1,2), P<0.001]. In terms of evaluating the unsuitability of medication, the proportion of patients with PIM in the polypharmacy group was higher than that in the non-polypharmacy group, including repeated medication [4.60% (5 227/113 594) vs. 1.64% (4 486/292 014)], contraindications [2.97% (3 376/113 594) vs. 1.13% (3 294/292 014)], interactions [6.51% (7 399/113 594) vs. 1.94% (5 658/292 014)], and lack of indications [22.39% (25 432/113 594) vs. 13.54% (39 543/292 014)], and the differences were all significant (all P<0.001). In terms of drug use categories, the top 5 most commonly prescribed drugs in the polypharmacy group were HMG-CoA reductase inhibitors (68 318 cases, 60.14%), dihydropyridines (60 951 cases, 53.66%), angiotensin receptor antagonists(45 050 cases, 39.66%), β-receptor blockers (25 675 cases, 22.60%) and sulfonylureas (16 023 cases, 14.11%). Conclusions:Polypharmacy is common in elderly patients with hypertension, diabetes mellitus, coronary artery disease, and cerebrovascular disease. The elderly patients with polypharmacy have a worse baseline status and more problems with PIM.

Epilepsy is a disorder of the brain charac-terized by abnormal neuron excitability.However,the underlying molecular mechanism of neuron excitability modulation remains elusive.With the help of bioinformatic methods,we have identified receptor-type tyrosine-pro-tein phosphatase-like N(PTPRN)as a critical gene dur-ing epileptogenesis.PTPRN recruits NEDD4L ubiquitin E3 ligase to NaV1.2 sodium channels,facilitating NEDD4L-mediated ubiquitination and endocytosis.Knockout of PTPRN endows hippocampal granule cells with augmented depolarization currents and higher intrinsic excitability,which is reflected by increased seizure susceptibility of transgenic mice.On the contrary,reduced neuron excit-ability and decreased seizure susceptibility are observed after PTPRN overexpression.Meanwhile,we find that a 133 aa fragment recaptures modulation effect of PTPRN full-length,and this fragment shows therapeutic potential towards epilepsy caused by NaV1.2 gain of function vari-ants.In brief,our results demonstrate PTPRN playsa criti-calroleinregulatingneuronexcitability,providing a poten-tial therapeutic approach for epilepsy.