Objective:To investigate the changes in hepatic phase II detoxification enzymes and their mechanism in metabolic associated steatohepatitis (MASH) induced by a methionine-choline-deficient (MCD) diet in mice.Methods:Ten C57BL/6J mice were randomly divided into two groups, with five mice in each group, and fed with a control diet (NCD group) and a methionine-choline-deficient diet (MCD group) for four consecutive weeks to establish the MASH model in mice. Mice body weight was recorded weekly. Mice peripheral blood and liver tissue samples were collected after four weeks. The liver histopathological changes were observed by hematoxylin-eosin staining and Sirius red staining in liver tissue. The levels of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglycerides were measured by an automatic biochemical analyzer. Triglyceride and total cholesterol were used to evaluate the lipid accumulation condition in the liver of mice with Oil red O staining. Real-time fluorescence quantitative PCR was used to detect the expression of liver inflammatory factors interleukin (IL)-1β and monocyte chemoattractant protein-1 (MCP-1) condition. Transcriptome sequencing and bioinformatics were used to analyze the changes in gene expression profiles in the liver of mice and screen differentially expressed genes. The expression conditions of phase Ⅱ detoxification enzymes glutathione S-transferase mu 4 (GSTM4), dihydronicotinamide riboside:quinone oxidoreductases (NQO-2), sulfotransferase 1β1 (SULT1β1), and uridine diphosphate glucuronosyltransferase 2 family, polypeptide A3(UGT2A3) were verified by real-time fluorescent quantitative PCR. Plasma malondialdehyde content, total antioxidant capacity (T-AOC), plasma and liver glutathione content were determined using commercial kits. The expression of nuclear factor E2-related factor 2 (Nrf2), GSTM4, and UGT1A6 was examined by Western blotting. The independent sample t-test was used for comparison between the groups. Results:The body weight of mice in the MCD group showed a gradual downward trend, while the body weight of mice in the NCD group did not change significantly following four weeks of different dietary feeding. The MCD group mice liver had yellow-white appearance with round edges. The liver/body mass index was significantly lower in the NCD group ( t=3.216, P<0.01). Hematoxylin-eosin staining showed that hepatocytes in the MCD group had an occurrence of fatty degeneration accompanied by inflammatory cell infiltration, with a higher NAFLD activity score (NAS) compared to the NCD group ( t=7.155, P<0.001). Sirius red staining showed that the the liver of the MCD group had mildly increased periportal fibers. Plasma biochemical tests indicated that plasma ALT, AST, and triglyceride levels were significantly higher in the MCD group than those in the NCD group ( t=8.920, P<0.001; t=6.696, P<0.001; t=3.904, P<0.01). Oil red O staining showed that a large number of lipid droplets accumulated in the liver tissue of the MCD group and were more severe than those in the NCD group ( t=7.405, P<0.001). The triglyceride content was significantly higher in the liver of the mice in the MCD group than that in the NCD group ( t=3.559, P<0.01), and the expression of inflammatory factors IL-1β and MCP-1 was significantly increased ( t=2.562 and 2.391, respectively, P<0.05). Transcriptome sequencing analysis showed that the expression profile of genes related to lipid metabolism was changed in the liver tissue of the mice in the MCD group. The expression of multiple phase Ⅱ detoxification enzymes was significantly downregulated. Real-time fluorescence quantitative PCR verification demonstrated that the expression of four phase Ⅱ detoxification enzymes GSTM4, NQO2, SUIL1β1, and UGT2A3 were significantly lower in the liver of the mice in the MCD group than those in the NCD group ( t=2.498, 3.570, 3.768, and 4.166, respectively, P<0.05). The detection kit showed that compared with the NCD group, the malondialdehyde content in the liver of mice in the MCD group increased ( t=3.601, P<0.01), while the plasma total glutathione ( t=11.93, P<0.001) and reduced glutathione levels were significantly reduced ( t=3.635, P<0.01). The total antioxidant capacity of the liver decreased ( t=2.872, P<0.05), and the total glutathione and reduced glutathione levels in the liver were significantly increased ( t=3.175 and 3.064, P<0.05). Western blotting showed that the expression of Nrf2, GSTM4, and UGT1A6 proteins was significantly lower in the MCD group than that in the NCD group ( t=3.385, 2.990, 2.168, P<0.05). Conclusions:The expressions of multiple phase Ⅱ detoxification enzymes and antioxidant capacity are reduced in the liver of MASH mice induced by the MCD diet, and its mechanism is related to the down-regulation of the expression of the upstream regulatory factor Nrf2 protein.

Objective·To construct a nanoengineered T cell system and explore its synergistic efficacy in the in vitro treatment of oral squamous cell carcinoma,and provide a new strategy for this disease.Methods·Zr-MOF nanoparticles with a highly hydrophilic hyaluronic acid(HA)coating(PCN224-HA NPs)were prepared,and their carboxyl groups were activated to enable coupling with anti-CD45 antibodies through an amide reaction.High-purity CD8+T cells were then extracted from the spleens of mice,and the PCN224-HA NPs were loaded onto the T cells via the CD45 antigens on their surface.The coupling was observed using bioelectron microscopy and confocal microscopy,and free radical generation was detected under ultrasound irradiation.The viability of Cal27 cells was recorded by a microplate reader to explore therapeutic effects in vitro.Results·PCN224-HA NPs were successfully synthesized and confirmed to be coupled onto the surface of CD8+T cells,constructing a nanoengineered T cell system(T cells@PCN224-HA NPs).Free radical production was monitored after ultrasound irradiation and the viability of Cal27 cells decreased to 31.70%after in vitro treatment.Conclusions·Preliminary experiments demonstrated that the nanoparticles can be successfully loaded onto the surface of T cells,providing combined sonodynamic therapeutic effects from the nanoparticles and immunocidal effects from the T cells.This offers a novel strategy for the treatment of oral squamous cell carcinoma,with potential for further in vivo anti-tumor experiments and therapeutic applications.

Objective·To construct and validate a nomogram for distinguishing primary Sj?gren's syndrome(PSS)from immunoglobulin G4-related sialadenitis(IgG4-RS)based on ultrasound scoring parameters and clinical indicators.Methods·A total of 141 patients with PSS and 31 patients with IgG4-RS were retrospectively recruited from Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine from January 2018 to December 2023.The ultrasound scoring parameters,including parotid gland ultrasound(PGUS)score,submandibular gland ultrasound(SMGUS)score,and salivary gland ultrasound(SGUS)score,along with clinical indicators such as gender,age,anti-SSB/La antibody,anti-SSA/Ro60 antibody,anti-SSA/Ro52 antibody,IgG,and rheumatoid factor(RF),were collected.The optimal US scoring parameters and clinical indicators were screened by least absolute shrinkage and selection operator(LASSO)regression,and a nomogram model was constructed to distinguish PSS from IgG4-RS.The internal validation of the model was carried out through bootstrap method.Receiver operator characteristic(ROC)curve,calibration curve and decision curve analysis(DCA)were used to estimate the discrimination,calibration,and clinical utility of the nomogram model,respectively.Results·LASSO regression identified six major variables:gender,age,anti-SSA/Ro60 antibody,anti-SSA/Ro52 antibody,PGUS score,and SMGUS score.These variables were used to construct the nomogram.ROC curve of the nomogram showed that the area under the curve(AUC)was 0.976,indicating the nomogram had strong discrimination ability.The bootstrap method was used for internal validation with 1 000 resampling iterations,and the average absolute error was 0.018.Calibration curve demonstrated good agreement between predicted and observed values.DCA indicated that the nomogram had certain clinical utility.Conclusion·The nomogram based on ultrasound scoring parameters and clinical indicators demonstrates excellent discrimination and calibration in differentiating PSS from IgG4-RS.It has the potential to assist in clinical diagnosis and decision-making.

Objective·To construct a nanoengineered T cell system and explore its synergistic efficacy in the in vitro treatment of oral squamous cell carcinoma,and provide a new strategy for this disease.Methods·Zr-MOF nanoparticles with a highly hydrophilic hyaluronic acid(HA)coating(PCN224-HA NPs)were prepared,and their carboxyl groups were activated to enable coupling with anti-CD45 antibodies through an amide reaction.High-purity CD8+T cells were then extracted from the spleens of mice,and the PCN224-HA NPs were loaded onto the T cells via the CD45 antigens on their surface.The coupling was observed using bioelectron microscopy and confocal microscopy,and free radical generation was detected under ultrasound irradiation.The viability of Cal27 cells was recorded by a microplate reader to explore therapeutic effects in vitro.Results·PCN224-HA NPs were successfully synthesized and confirmed to be coupled onto the surface of CD8+T cells,constructing a nanoengineered T cell system(T cells@PCN224-HA NPs).Free radical production was monitored after ultrasound irradiation and the viability of Cal27 cells decreased to 31.70%after in vitro treatment.Conclusions·Preliminary experiments demonstrated that the nanoparticles can be successfully loaded onto the surface of T cells,providing combined sonodynamic therapeutic effects from the nanoparticles and immunocidal effects from the T cells.This offers a novel strategy for the treatment of oral squamous cell carcinoma,with potential for further in vivo anti-tumor experiments and therapeutic applications.

Prescription sequence symmetry analysis (PSSA) is one of the important methods for post-marketing pharmacovigilance based on the real-world medical prescription databases. It can be used to detect prescription cascades and mine adverse drug reaction (ADR) signals, which has been verified by many studies. PSSA shows high specificity and medium sensitivity in identifying ADR. It can quantify the correlation or risks of ADR. It is easy to use and simple in algorithm, and it has good robustness to some non time-dependent confounding factors. However, the results may be affected by some human confounding factors and data quality. This paper reviews the principle, calculation method, application scope, and precaution of PSSA by reviewing related literature on PSSA domestically and abroad, in order to provide reference for pharmacovigilance in China.

Objective:To understand the application situation and role of prescription sequence symmetry analysis (PSSA) in pharmacovigilance.Methods:The relevant databases at home and abroad were searched (up to April 30, 2024), and the original articles using PSSA as the research method were collected. The basic information of the literature (first author, publication year, country, etc.), the purpose and main content of the study, the index drugs as well as the marker drugs or medical diagnoses involved in the adverse drug reactions (ADRs) were extracted. Descriptive statistical analysis was carried out.Results:A total of 66 articles were included in the analysis. The first article was published in 1996, the number of articles published in recent years has increased significantly, and those published after 2016 accounted for 68.2% (45/66). The top 3 countries in terms of published literature quantity were the United States, Denmark, and Japan. The index drugs most commonly studied were those for the cardiovascular system and the neuropsychiatric system, in 18 and 14 articles respectively. The drugs studied in 3 or more papers were hypolipidemic drugs, antihypertensive drugs, antipsychotics, antiepileptics, proton pump inhibitors, hypoglycemic drugs and anticoagulants. The targeted ADRs/diseases most studied were those about the neuropsychiatric system (in 13 studies), followed by those about the endocrine and metabolic system (in 12 studies). The research objective in 47 articles was to explore the association between index drugs and ADRs/diseases through PSSA. Finally, the associations between 21 ADRs and index drugs were identified in 24 articles, of which 9 were new ADRs not recorded in drug instructions; benefits or potential preventive and therapeutic effects of index drugs on certain diseases were found in 7 studies. Ten studies were conducted to explore ADR information of specific drugs or detect suspicious drugs that cause specific ADRs, and some correlation signals between drugs and ADRs that previously unknown were detected. Nine studies evaluated the prescribing cascades, including the use of antitussive drugs after ACEI, the prescribing cascades related to drug-induced lower urinary tract symptoms and edema, the prescription cascades of statins, and the prescribing cascade relic.Conclusion:PSSA is a useful method for identifying potential prescribing cascades and mining ADR signals using medical prescription databases, especially suitable for the safety monitoring of long-term medication for chronic diseases and the signal detection of ADR that causal relationships are difficult to determine.

Prescription sequence symmetry analysis (PSSA) is one of the important methods for post-marketing pharmacovigilance based on the real-world medical prescription databases. It can be used to detect prescription cascades and mine adverse drug reaction (ADR) signals, which has been verified by many studies. PSSA shows high specificity and medium sensitivity in identifying ADR. It can quantify the correlation or risks of ADR. It is easy to use and simple in algorithm, and it has good robustness to some non time-dependent confounding factors. However, the results may be affected by some human confounding factors and data quality. This paper reviews the principle, calculation method, application scope, and precaution of PSSA by reviewing related literature on PSSA domestically and abroad, in order to provide reference for pharmacovigilance in China.

Objective:To understand the application situation and role of prescription sequence symmetry analysis (PSSA) in pharmacovigilance.Methods:The relevant databases at home and abroad were searched (up to April 30, 2024), and the original articles using PSSA as the research method were collected. The basic information of the literature (first author, publication year, country, etc.), the purpose and main content of the study, the index drugs as well as the marker drugs or medical diagnoses involved in the adverse drug reactions (ADRs) were extracted. Descriptive statistical analysis was carried out.Results:A total of 66 articles were included in the analysis. The first article was published in 1996, the number of articles published in recent years has increased significantly, and those published after 2016 accounted for 68.2% (45/66). The top 3 countries in terms of published literature quantity were the United States, Denmark, and Japan. The index drugs most commonly studied were those for the cardiovascular system and the neuropsychiatric system, in 18 and 14 articles respectively. The drugs studied in 3 or more papers were hypolipidemic drugs, antihypertensive drugs, antipsychotics, antiepileptics, proton pump inhibitors, hypoglycemic drugs and anticoagulants. The targeted ADRs/diseases most studied were those about the neuropsychiatric system (in 13 studies), followed by those about the endocrine and metabolic system (in 12 studies). The research objective in 47 articles was to explore the association between index drugs and ADRs/diseases through PSSA. Finally, the associations between 21 ADRs and index drugs were identified in 24 articles, of which 9 were new ADRs not recorded in drug instructions; benefits or potential preventive and therapeutic effects of index drugs on certain diseases were found in 7 studies. Ten studies were conducted to explore ADR information of specific drugs or detect suspicious drugs that cause specific ADRs, and some correlation signals between drugs and ADRs that previously unknown were detected. Nine studies evaluated the prescribing cascades, including the use of antitussive drugs after ACEI, the prescribing cascades related to drug-induced lower urinary tract symptoms and edema, the prescription cascades of statins, and the prescribing cascade relic.Conclusion:PSSA is a useful method for identifying potential prescribing cascades and mining ADR signals using medical prescription databases, especially suitable for the safety monitoring of long-term medication for chronic diseases and the signal detection of ADR that causal relationships are difficult to determine.

Objective·To construct and validate a nomogram for distinguishing primary Sj?gren's syndrome(PSS)from immunoglobulin G4-related sialadenitis(IgG4-RS)based on ultrasound scoring parameters and clinical indicators.Methods·A total of 141 patients with PSS and 31 patients with IgG4-RS were retrospectively recruited from Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine from January 2018 to December 2023.The ultrasound scoring parameters,including parotid gland ultrasound(PGUS)score,submandibular gland ultrasound(SMGUS)score,and salivary gland ultrasound(SGUS)score,along with clinical indicators such as gender,age,anti-SSB/La antibody,anti-SSA/Ro60 antibody,anti-SSA/Ro52 antibody,IgG,and rheumatoid factor(RF),were collected.The optimal US scoring parameters and clinical indicators were screened by least absolute shrinkage and selection operator(LASSO)regression,and a nomogram model was constructed to distinguish PSS from IgG4-RS.The internal validation of the model was carried out through bootstrap method.Receiver operator characteristic(ROC)curve,calibration curve and decision curve analysis(DCA)were used to estimate the discrimination,calibration,and clinical utility of the nomogram model,respectively.Results·LASSO regression identified six major variables:gender,age,anti-SSA/Ro60 antibody,anti-SSA/Ro52 antibody,PGUS score,and SMGUS score.These variables were used to construct the nomogram.ROC curve of the nomogram showed that the area under the curve(AUC)was 0.976,indicating the nomogram had strong discrimination ability.The bootstrap method was used for internal validation with 1 000 resampling iterations,and the average absolute error was 0.018.Calibration curve demonstrated good agreement between predicted and observed values.DCA indicated that the nomogram had certain clinical utility.Conclusion·The nomogram based on ultrasound scoring parameters and clinical indicators demonstrates excellent discrimination and calibration in differentiating PSS from IgG4-RS.It has the potential to assist in clinical diagnosis and decision-making.

Objective:To investigate the changes in hepatic phase II detoxification enzymes and their mechanism in metabolic associated steatohepatitis (MASH) induced by a methionine-choline-deficient (MCD) diet in mice.Methods:Ten C57BL/6J mice were randomly divided into two groups, with five mice in each group, and fed with a control diet (NCD group) and a methionine-choline-deficient diet (MCD group) for four consecutive weeks to establish the MASH model in mice. Mice body weight was recorded weekly. Mice peripheral blood and liver tissue samples were collected after four weeks. The liver histopathological changes were observed by hematoxylin-eosin staining and Sirius red staining in liver tissue. The levels of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglycerides were measured by an automatic biochemical analyzer. Triglyceride and total cholesterol were used to evaluate the lipid accumulation condition in the liver of mice with Oil red O staining. Real-time fluorescence quantitative PCR was used to detect the expression of liver inflammatory factors interleukin (IL)-1β and monocyte chemoattractant protein-1 (MCP-1) condition. Transcriptome sequencing and bioinformatics were used to analyze the changes in gene expression profiles in the liver of mice and screen differentially expressed genes. The expression conditions of phase Ⅱ detoxification enzymes glutathione S-transferase mu 4 (GSTM4), dihydronicotinamide riboside:quinone oxidoreductases (NQO-2), sulfotransferase 1β1 (SULT1β1), and uridine diphosphate glucuronosyltransferase 2 family, polypeptide A3(UGT2A3) were verified by real-time fluorescent quantitative PCR. Plasma malondialdehyde content, total antioxidant capacity (T-AOC), plasma and liver glutathione content were determined using commercial kits. The expression of nuclear factor E2-related factor 2 (Nrf2), GSTM4, and UGT1A6 was examined by Western blotting. The independent sample t-test was used for comparison between the groups. Results:The body weight of mice in the MCD group showed a gradual downward trend, while the body weight of mice in the NCD group did not change significantly following four weeks of different dietary feeding. The MCD group mice liver had yellow-white appearance with round edges. The liver/body mass index was significantly lower in the NCD group ( t=3.216, P<0.01). Hematoxylin-eosin staining showed that hepatocytes in the MCD group had an occurrence of fatty degeneration accompanied by inflammatory cell infiltration, with a higher NAFLD activity score (NAS) compared to the NCD group ( t=7.155, P<0.001). Sirius red staining showed that the the liver of the MCD group had mildly increased periportal fibers. Plasma biochemical tests indicated that plasma ALT, AST, and triglyceride levels were significantly higher in the MCD group than those in the NCD group ( t=8.920, P<0.001; t=6.696, P<0.001; t=3.904, P<0.01). Oil red O staining showed that a large number of lipid droplets accumulated in the liver tissue of the MCD group and were more severe than those in the NCD group ( t=7.405, P<0.001). The triglyceride content was significantly higher in the liver of the mice in the MCD group than that in the NCD group ( t=3.559, P<0.01), and the expression of inflammatory factors IL-1β and MCP-1 was significantly increased ( t=2.562 and 2.391, respectively, P<0.05). Transcriptome sequencing analysis showed that the expression profile of genes related to lipid metabolism was changed in the liver tissue of the mice in the MCD group. The expression of multiple phase Ⅱ detoxification enzymes was significantly downregulated. Real-time fluorescence quantitative PCR verification demonstrated that the expression of four phase Ⅱ detoxification enzymes GSTM4, NQO2, SUIL1β1, and UGT2A3 were significantly lower in the liver of the mice in the MCD group than those in the NCD group ( t=2.498, 3.570, 3.768, and 4.166, respectively, P<0.05). The detection kit showed that compared with the NCD group, the malondialdehyde content in the liver of mice in the MCD group increased ( t=3.601, P<0.01), while the plasma total glutathione ( t=11.93, P<0.001) and reduced glutathione levels were significantly reduced ( t=3.635, P<0.01). The total antioxidant capacity of the liver decreased ( t=2.872, P<0.05), and the total glutathione and reduced glutathione levels in the liver were significantly increased ( t=3.175 and 3.064, P<0.05). Western blotting showed that the expression of Nrf2, GSTM4, and UGT1A6 proteins was significantly lower in the MCD group than that in the NCD group ( t=3.385, 2.990, 2.168, P<0.05). Conclusions:The expressions of multiple phase Ⅱ detoxification enzymes and antioxidant capacity are reduced in the liver of MASH mice induced by the MCD diet, and its mechanism is related to the down-regulation of the expression of the upstream regulatory factor Nrf2 protein.