ObjectiveTo investigate the role and mechanism of action of paeoniflorin （PF） in protecting HepG2 cells induced by palmitic acid （PA）. MethodsHepG2 cells were stimulated with PA at a concentration of 250 μmol/L to establish a NAFLD model. Compound C at a concentration of 10 μmol/L was used as an inhibitor， and PF at a concentration of 25 μmol/L was used for intervention. The experiment was divided into normal group （CON group） treated with complete culture medium， model group （MOD group） treated with PA， PF treatment group （MOD+PF group） treated with PA and PF， model+inhibitor group （MOD+COM group） treated with PA and Compound C， and model+inhibitor+PF group （MOD+COM+PF group） treated with PA， Compound C， and PF. Kits were used to measure lipid deposition indicators， liver function parameters， oxidative stress indicators， and inflammation indicators； oil red O staining was used to observe lipid deposition； Western Blot was used to measure the protein expression levels of AMPK， SIRT1， PGC-1α， mTOR， Beclin-1， LC3， and P62 in cells. The one-way analysis of variance was used for comparison of quantitative data between groups， while the Tukey’s test was used for comparison between two groups. ResultsCompared with the MOD group， PF improved the levels of TC and TG （P<0.05）， reduced the levels of ALT， AST， CRP， TNF-α， IL-1β， and IL-6 （P<0.05）， increased the activity of SOD and CAT and the level of GSH， and reduced the level of MDA in cells （all P<0.05）. Oil red O staining showed that PF alleviated lipid deposition in cells. Western blot results showed that compared with the MOD group， PF increased the protein expression levels of p-AMPK， SIRT1， PGC-1α， LC3Ⅱ/LC3Ⅰ， and Beclin-1 and reduced the protein expression levels of p-mTOR and P62 （all P<0.05）. ConclusionPF can inhibit PA-induced oxidative stress and inflammatory response in HepG2 cells， improve lipid deposition， and promote autophagy via the AMPK/SIRT1/PGC-1α/mTOR signaling pathway.

ObjectiveTo develop a scale for evaluating the syndrome of internal retention of dampness and turbidity in nonalcoholic fatty liver disease （NAFLD） that combines disease and syndrome and has the characteristics of traditional Chinese medicine （TCM）. MethodsAn item pool was established for evaluating the syndrome of internal retention of dampness and turbidity in nonalcoholic fatty liver disease （NAFLD） with reference to the guideline for developing international scales. A clinical survey was conducted among the outpatients and inpatients who were diagnosed with NAFLD and had the syndrome of internal retention of dampness and turbidity in Department of Hepatology and Spleen-Stomach， The First Affiliated Hospital of Henan University of Chinese Medicine， from June to August， 2023， and the items were screened based on the classical test theory and the item response theory. An expert questionnaire was developed， and expert discussions were conducted using the Delphi method to identify the items for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD. Finally， the scale was given scientific scores. ResultsA preliminary item pool was established， with 16 primary items and 22 secondary items， and it was divided into the two dimensions of disease and syndrome type. Clinical pre-survey suggested to retain 9 primary items and 14 secondary items， while the Delphi expert questionnaire recommended to retain 11 primary items and 15 secondary items， and tongue manifestation and pulse manifestation were no longer used for assessing the severity of syndrome. After hierarchical analysis and scientific assignment of scores， the scale for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD had a total score of 123 points and 9 important items， i.e.， discomfort in the hypochondrium， abdominal fullness and distension， obesity， heaviness of the head and body， loose stool， anorexia， coughing up phlegm， nausea with a tendency to vomit， and lethargy. ConclusionA preliminary scale is established for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD， which fills the gap in this research field and provides a basis for further clinical application.

ObjectiveTo investigate the effect of modified Weijingtang on the pyroptosis of RAW264.7 macrophages via the cysteinyl aspartate-specific protease-1 （Caspase-1）/gasdermin D （GSDMD） pathway. MethodLipopolysaccharide （LPS） was used to induce pyroptosis of RAW264.7 cells. The blank group was treated with the blank serum， and the intervention groups were treated with the sera containing different doses of modified Weijingtang. After 24 h， the viability of cells in different groups was examined by the cell counting kit-8 （CCK-8）. The pyroptosis and morphology of cells in each group were observed by a scanning electron microscope and a phase-contrast microscope， respectively. The mRNA and protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）， Caspase-1， and GSDMD in each group were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The levels of interleukin （IL）-18 and IL-1β in each group were measured by enzyme-linked immunosorbent assay. ResultUnder the electron microscope， RAW264.7 cells presented the best morphology and structure in the blank group and obvious pyroptosis and leakage of cell contents in the model （LPS） group. Compared with the model group， the intervention groups showed reduced pyroptosis to varying degrees， and the high-dose group had the closest cell morphology and structure to the blank group. Under the optical microscope， RAW264.7 cells were spherical in the blank group and irregular with protrusions in the model group. Compared with the model group， the intervention groups showed improved cell morphology， and the cell morphology in the group with the dose of 20% was the closest to that in the blank group. The mRNA and protein levels of NLRP3， Caspase-1， and GSDMD in the model group were higher than those in the blank group （P<0.05）. Compared with the model group， each intervention group showed down-regulated expression of the above indicators （P<0.05）. Compared with the blank group， the model group presented elevated levels of IL-18 and IL-1β （P<0.05）， which were lowered in the intervention （10%， 20%） groups （P<0.01）. ConclusionModified Weijingtang inhibits the pyroptosis of macrophages by down-regulating the Caspase-1/GSDMD pathway and reducing the release of proinflammatory cytokines.

Objective To investigate the risk factors for the development of advanced liver fibrosis by analyzing the clinical features of patients with in nonalcoholic fatty liver disease(NAFLD)and advanced liver fibrosis,and to establish a nomogram model for predicting the risk of advanced liver fibrosis.Methods A retrospective analysis was performed for the clinical data of 406 NAFLD patients who attended The First Affiliated Hospital of Henan University of Chinese Medicine from January 2022 to October 2023,and according to whether liver stiffness measurement(LSM)measured by FibroScan was≥11.0 kPa,the patients were divided into advanced liver fibrosis group with 65 patients and non-advanced liver fibrosis group with 341 patients.Related data were collected,including general information,laboratory markers,and medical history.The independent-samples t test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups;the chi-square test was used for comparison of categorical data between two groups.A multivariate Logistic regression analysis was used to identify independent risk factors,and a nomogram model was established based on these factors.The receiver operating characteristic(ROC)curve was used to evaluate the discriminatory ability of the nomogram model,and the calibration curve was used to evaluate its effectiveness.Results The univariate analysis showed that there were significant differences between the advanced liver fibrosis group and the non-advanced liver fibrosis group in age,controlled attenuation parameter(CAP),total bilirubin,direct bilirubin(DBil),indirect bilirubin,globin,alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase(ALP),gamma-glutamyl transpeptidase(GGT),glucose,body mass index(BMI),and history of diabetes(all P<0.05).The multivariate Logistic regression analysis showed that CAP(odds ratio[OR]=1.015,95%confidence interval[CI]:1.006-1.024,P=0.010),DBil(OR=1.345,95%CI:1.139-1.590,P=0.001),ALP(OR=1.019,95%CI:1.008-1.029,P=0.001),GGT(OR=1.004,95%CI:1.000-1.008,P=0.035)and BMI(OR=1.240,95%CI:1.137-1.353,P=0.001)were independent risk factors for the development of advanced liver fibrosis in NAFLD.A nomogram model was established based on the results of the multivariate Logistic regression analysis.The ROC curve analysis showed that this nomogram model had an area under the ROC curve of 0.841(95%CI:0.791-0.891)in predicting the development of advanced liver fibrosis in the NAFLD population,and the calibration curve showed a good degree of fitting between the observed and predicted values for the development of advanced liver fibrosis.Conclusion Elevated levels of CAP,BMI,DBil,ALP,and GGT are independent risk factors for advanced liver fibrosis in NAFLD.The nomogram model established based on these factors has good predictive performance and a certain value in predicting advanced liver fibrosis.

Endodontic diseases are a kind of chronic infectious oral disease.Common endodontic treatment concepts are based on the removal of inflamed or necrotic pulp tissue and the replacement by gutta-percha.However,it is very essential for endodontic treatment to debride the root canal system and prevent the root canal system from bacterial reinfection after root canal therapy(RCT).Recent research,encompassing bacterial etiology and advanced imaging techniques,contributes to our understanding of the root canal system's anatomy intricacies and the technique sensitivity of RCT.Success in RCT hinges on factors like patients,infection severity,root canal anatomy,and treatment techniques.Therefore,improving disease management is a key issue to combat endodontic diseases and cure periapical lesions.The clinical difficulty assessment system of RCT is established based on patient conditions,tooth conditions,root canal configuration,and root canal needing retreatment,and emphasizes pre-treatment risk assessment for optimal outcomes.The findings suggest that the presence of risk factors may correlate with the challenge of achieving the high standard required for RCT.These insights contribute not only to improve education but also aid practitioners in treatment planning and referral decision-making within the field of endodontics.

Objective:A SARS-CoV-2 pseudovirus(PsV)system was established for neutralizing antibody evaluation and virus entry inhibitor screening.Methods:Lentiviral vector plasmids psPAX2,pCDH-Luc and SARS-CoV-2 Spike(S)protein expres-sion plasmids were co-transfected,and harvested pseudoviral supernatant was used to infect ACE2-293T cells.Protein content of p24 was determined to reflect titer of PsV,and expression of S protein in PsV was detected by Western blot.Neutralization capacity of an S protein monoclonal antibody was evaluated using original strain,D614G,Gamma,Delta,Omicron PsV.Two reported virus entry inhibitors,chloroquine and carrageenin,were used to detect effect on entry of Omicron PsV.Results:Lentiviral vector successfully incorporated S protein.Western blot results showed that S protein mutated at 665Y showed a different cleavage form(90 kD)than wild-type full-length S protein(180 kD).Titer of PsV packaged by three plasmids system was higher.Ratio of S protein expression plasmid,transfer plasmid and packaging plasmid at 1∶3∶3 was optimum condition for viral packaging.Titer of PsV packaged under this condi-tion was over 20 ng/ml.PsV could effectively infect ACE2-293T cells,and double reporter gene GFP and firefly luciferase were expressed obviously,whose chemiluminescence values reached 106.Monoclonal antibodies of S protein effectively neutralized four types of PsVs,but neutralization of original strain was 10～30 times greater than that of variant PsV.Virus entry inhibitors,chloroquine and ι-carrageenan significantly inhibited entry of Omicron PsV.Conclusion:SARS-CoV-2 PsV infection system we conducted can simu-late entry of SARS-CoV-2 successfully.Effective pharmacodynamic evaluation of neutralizing antibodies and virus entry inhibitors can be performed efficiently by the system,which can provide a technical platform for evaluation of neutralizing antibody of SARS-CoV-2 and screening of virus entry inhibitors,and would benefit R&D of anti-SARS-CoV-2 drugs.

Nonalcoholic fatty liver disease (NAFLD) has gradually become a prominent cause affecting human liver health, and the development and progression of NAFLD are associated with metabolic dysfunction, with glucose and lipid metabolism disorder as the key link in this process. Takeda G protein-coupled receptor 5 (TGR5) is one of the main receptors of bile acid and is extensively expressed in the body, and glucose and lipid metabolism mediated by TGR5 plays an important role in the human body. This article summarizes the role and mechanism of TGR5 in glucose and lipid metabolism and the research findings of the treatment of NAFLD based on TGR5, in order to provide a reference for basic and clinical research.

Nonalcoholic fatty liver disease (NAFLD) has been renamed as metabolic-associated fatty liver disease, and systemic metabolic dysfunction has become one of the concerns of this disease. NAFLD is a metabolic disease based on dyslipidemia in the liver, which is closely associated with adipose tissue. Hepatokines and adipokines secreted by the liver and adipose tissue play an important role in regulating liver lipid metabolism. This article summarizes the hepatokines and adipokines that can promote or inhibit lipid metabolism, focusing on the mechanism of lipid metabolism mediated by hepatokines and adipokines in NAFLD, so as to provides ideas and a theoretical basis for clinical prevention and treatment.

Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease in which a large amount of fat accumulates in hepatocytes due to lipid metabolism disorders. Conventional anti-inflammatory and transaminase-lowering treatment regimens often have an unsatisfactory therapeutic effect, and restoring the normal biosynthesis and metabolism of lipids is the key to the treatment of NAFLD. Studies have shown that brown adipose tissue can improve metabolic diseases by enhancing insulin sensitivity and regulating lipid metabolism, and the treatment of NAFLD by promoting white fat browning has attracted wide attention in the medical field. This article reviews the mechanism of white fat browning in improving NAFLD and summarizes the hepatokines that can promote white fat browning, so as to provide new ideas for the clinical treatment of NAFLD.

Total knee arthroplasty， as a common treatment option for advanced knee osteoarthritis， can alleviate the clinical symptoms of patients. Deep vein thrombosis of the lower extremity is the most common complication of total knee arthroplasty. Previous studies have shown that the occurrence of deep vein thrombosis after total knee arthroplasty is mostly related to the overexpression of inflammatory factors in vivo. Nuclear transcription factor-κB， Toll-like receptor 4， phosphatidylinositol 3-kinase/protein kinase B， tumor necrosis factor， and nuclear transcription factor E₂-related factor 2 are typical signaling pathways related to inflammation. Regulating the expression of the signaling pathways can intervene the formation of inflammatory factors. Inhibiting the formation of inflammatory factors can help suppress the activation of platelets， thereby blocking thrombosis. According to previous research， Chinese medicine monomers， Chinese medicine extract， and compound Chinese medicine prescriptions all directly or indirectly inhibit the expression of inflammatory factors by regulating the above signaling pathways， thereby suppressing the occurrence of deep vein thrombosis after total knee arthroplasty. Therefore， Chinese medicine can reduce postoperative complications and promote postoperative recovery of patients at low cost with small side effects. This article summarizes the research on Chinese medicine intervention on deep vein thrombosis-related signaling pathways after total knee arthroplasty， which is expected to lay a basis for the in-depth study and clinical application of Chinese medicine in deep vein thrombosis.