ObjectiveTo compare the hepatic bile acid profile between a mouse model of metabolic-associated steatohepatitis （MASH） induced by a high-fat， high-sugar， and high-cholesterol diet combined with intraperitoneal injection of 10% carbon tetrachloride （CCl₄） and MASH cases in clinical practice， and to investigate the feasibility of this model in studying drug interventions on bile acid profile in MASH. MethodsA total of 30 male C57BL/6J mice were randomly divided into control group and model group， with 15 mice in each group. The mice in the control group were given normal diet and drinking water and weekly injections of olive oil， and those in the model group were given a high-fat， high-sugar， and high-cholesterol diet， high-sugar drinking water， and weekly injections of CCl₄+olive oil. At the end of weeks 8， 12， and 16， 5 mice were selected from each group to collect samples. Behavioral assessments were performed， and body weight and liver wet weight were measured； liver pathology and lipid deposition were evaluated by HE staining， SAF scoring， oil Red O staining， the semi-quantitative analysis of stained area， the serum levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， and liver triglyceride （TG） content； Sirius Red staining was performed for liver tissue to assess liver fibrosis； ultra-performance liquid chromatography-tandem mass spectrometry and targeted metabolomics were used to measure the hepatic bile acid profile， including cholic acid （CA）， glycocholic acid （GCA）， chenodeoxycholic acid （CDCA）， glycochenodeoxycholic acid （GCDCA）， ursodeoxycholic acid （UDCA）， tauroursodeoxycholic acid （TUDCA）， hyodeoxycholic acid （HDCA）， and glycodeoxycholic acid （GDCA）. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsCompared with the control group at the same time point， the model group had disheveled and dull fur， reduced activity， and relatively slow reactions at weeks 8， 12， and 16， as well as significant increases in liver wet weight （P<0.05）， the serum level of ALT （P<0.05）， the content of TG in the liver （P<0.05）， and SAF score （P<0.05）. As for the differentially expressed bile acids in liver tissue， compared with the control group at week 8， the model group had significantly higher levels of CA and CDCA and significantly lower levels of UDCA， TUDCA， HDCA， and GDCA （all P<0.05）； compared with the control group at week 12， the model group had significantly higher levels of CA， GCA， CDCA， and GCDCA and significantly lower levels of UDCA and HDCA （all P<0.05）； compared with the control group at week 16， the model group had significantly higher levels of CA， GCA， CDCA， GCDCA， and TUDCA and significantly lower levels of UDCA， HDCA， and GDCA （all P<0.05）. As for the differentially expressed bile acids in the bile acid pool of liver tissue， compared with the control group at week 8， the model group had significantly higher levels of CA and CDCA and significantly lower levels of UDCA， TUDCA， GDCA， and HDCA （all P<0.05）； compared with the control group at weeks 12 and 16， the model group had significantly higher levels of GCA and GCDCA and significantly lower levels of UDCA， GDCA， and HDCA （all P<0.05）. ConclusionThere are significant changes in the hepatic bile acid profile in a mouse model of MASH induced by a high-fat， high-sugar， and high-cholesterol diet combined with CCl₄， which are similar to the changes in bile acids in MASH cases in clinical practice， suggesting that this model can be used to explore the interventional effect of drugs on the bile acid profile in MASH.

ObjectiveTo explore the mechanism and pathway of Gandou Fumu decoction （GDFMD） in the development of liver fibrosis in Wilson's disease （WD）. MethodFirst， 30 TX-j mice were randomly divided into the model group， high-dose， medium-dose， and low-dose GDFMD groups， and penicillamine group， with six mice in each group， and another six wild-type mice were used as the normal group. The high-dose， medium-dose， and low-dose GDFMD groups were intragastrically administered drugs of 13.92， 6.96， 3.48 g·kg^-1. In the penicillamine group， 0.1 g·kg^-1 of penicillamine was given by intragastric administration. The model group and the normal group were given equal volume of normal saline， once a day， for four consecutive weeks. Samples were collected four weeks after gavage， and enzyme-linked immunosorbent assay （ELISA） was used to detect type Ⅲ procollagen peptide （PCⅢ）， collagen type Ⅳ （Col Ⅳ）， hyaluronic acid （HA）， and laminin （LN）. Hematoxylin-eosin （HE）， Masson， and picric acid-Sirus red collagen （Sirus Red） staining were used to observe the histopathological changes of liver fibrosis. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， immunohistochemistry， and Western blot were used to observe the expressions of α-smooth muscle actin （α-SMA） and collagen type Ⅰ （Col Ⅰ）， which were related to the activation of hepatic stellate cells （HSCs）. The expression of miR-29b-3p was observed by Real-time PCR. The expression of Unc-51-like kinase 1 （ULK1） and its downstream-related factors were observed by Western blot. The downstream genes of miR-29b-3p were verified by the dual luciferase reporter gene detection method. ResultCompared with the normal group， the four items of liver fibrosis （PCⅢ， Col Ⅳ， HA， and LN） in the model group were significantly abnormal （P<0.01）， and the pathology was significantly abnormal. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ， as well as α-SMA mRNA and Col Ⅰ mRNA was up-regulated （P<0.05， P<0.01）， and miR-29b-3p expression was down-regulated （P<0.01）. ULK1， p-ULK1， autophagy-related gene 13 （Atg13）， p-Atg13， Beclin-1， FAK family kinase-interacting protein of 200 kDa （FIP200）， activating molecule in BECN1-regulated autophagy protein 1 （AMBKA1）， and microtubule-associated protein 1 light chain 3Ⅱ/Ⅰ（LC3Ⅱ/Ⅰ） were up-regulated （P<0.05， P<0.01）. p62 protein expression was down-regulated （P<0.01）. Compared with the model group， the four items of liver fibrosis in the high-dose， medium-dose， and low-dose GDFMD groups and the penicillamine group were significantly improve （P<0.01）， and the pathological conditions were improved. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ， as well as α-SMA mRNA and Col Ⅰ mRNA was down-regulated （P<0.05， P<0.01）， and the expression of miR-29b-3p was up-regulated （P<0.01）. ULK1， p-ULK1， Atg13， p-Atg13， Beclin-1， FIP200， AMBKA1， and LC3Ⅱ/Ⅰ were down-regulated （P<0.05， P<0.01）， and p62 protein expression was up-regulated （P<0.01）. The prediction software predicted that there was a binding site between miR-29b-3p and ULK1. The dual-luciferase reporter gene detection method indicated that the luciferase activity of the ULK1-WT plasmid-transfected cell group was reduced when miR-29b-3p mimics were co-cultured （P<0.01）. ConclusionGDFMD can regulate ULK1-mediated autophagy by up-regulating miR-29b-3p and further exert its anti-hepatic fibrosis effect in Wilson's disease.

Objective To study the levels and clinical significance of serum angiopoietin-like protein 8(ANGPTL8)and neopurine in children with hormone sensitive primary nephrotic syndrome(NS).Methods A total of 159 children with hormone sensitive primary NS treated in the hospital from January 2018 to January 2021 were selected as the study subjects(NS group).After 1 year of follow-up,based on whether frequent recurrence occurred,they were divided into the non frequent recurrence subgroup(n=93)and the frequent recurrence subgroup(n=66).Additionally,60 children with oblique hernia who underwent e-lective surgery in the Department of Pediatrics in the hospital were selected as the control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of ANGPTL8 and neopterin.The ser-um levels of ANGPTL8 and neopterin in each group were compared.Multivariate Logistic regression analysis was used to analyze risk factors for frequent recurrence in children with hormone sensitive primary NS.The predictive value of serum ANGPTL8 and neopterin in predicting frequent recurrence in children with hormone sensitive primary NS was analyzed by receiver operating characteristic(ROC)curve.Results The serum lev-els of ANGPTL8 and neopterin in the NS group were higher than those in the control group,and the differ-ences were statistically significant(t=20.948,44.288,P＜0.001,0.001).The serum ANGPTL8,neopterin levels,CD8+T cells,24 h urine protein quantification,and urine protein conversion time in the frequent recur-rence subgroup were significantly higher than those in the non frequent recurrence subgroup,while the blood albumin,CD4+T cells,CD4+/CD8+T cell ratios were lower than those in the non frequent recurrence sub-group,and the differences were statistically significant(all P＜0.05).The serum levels of ANGPTL8 and neopterin in NS patients were significantly positively correlated with 24 h urine protein and CD8+T cells,while they were significantly negatively correlated with serum albumin,CD4+T cells,CD4l+/CD8+(all P＜0.05).Serum ANGPTL8 and neopterin were independent risk factors for frequent recurrence in children with hormone sensitive primary NS.The area under the curve(AUC)of the combined model of serum ANGPTL8 and neopurine for predicting frequent recurrence in hormone sensitive primary NS patients was 0.852(95％CI:0.813-0.889),which was higher than 0.764(95％CI:0.722-0.812)of ANGPTL8,and 0.749(95％CI:0.711-0.790)of neopurine,and the differences were statistically significant(Z=3.623,3.987,P=0.003,＜0.001).Conclusion Elevated levels of serum ANGPTL8 and neopterin are risk factors for frequent recurrence hormone sensitive primary NS.The combination detection of serum ANGPTL8 and neopterin has high predictive value for frequent recurrence in children with hormone sensitive primary NS.

Objective To conduct lipidomics analysis of medical Marmota himalayana oil and explore its potential biological effects. Methods Marmota himalayana abdominal fat was heated and refined to produce medical marmot oil. Lipidomics analysis of two batches of Marmota himalayana oil was performed using gas chromatography-mass spectrometry (GC-MS) and ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS). The potential effects of Marmota himalayana oil were analyzed based on the results of lipid composition detection. Results GC-MS results showed that the main components of the two batches of Marmota himalayana oil were saturated fatty acids (volume fraction of about 60%) and unsaturated fatty acids (volume fraction of about 30%). The saturated fatty acids were mainly palmitic acid, stearic acid, and arachidic acid, while the unsaturated fatty acids were mainly oleic acid and linoleic acid. There were no significant differences in the volume fraction of various fatty acids between the two batches, indicating that the preparation process of Marmota himalayana oil was stable. The results of UPLC-MS negative ion mode detection showed that Marmota himalayana oil contained 15 saturated fatty acids and 18 unsaturated fatty acids, but there were significant differences in volume fraction between the two batches. The results of UPLC-MS positive ion mode detection showed that Marmota himalayana oil of both batches contained 15 diglycerides and 30 triglycerides. Conclusion This study innovatively applies GC-MS and UPLC-MS techniques to analyze the lipidomic components of medical Marmota himalayana oil, providing a reliable basis for subsequent exploration of its pharmacological activity and the establishment of strict quality control standards.

Osteochondral lesion of talus (OLT) is a foot and ankle disease characterized by ankle pain, which may impact the joint function and life quality. If managed improperly, it may lead to a further ankle arthritis, severely compromising the prognosis. The therapeutic effect of conservative treatment for OLT is still uncertain. Surgery is still the main treatment modality for OLT with various techniques. However, the optimized surgical technique is still inconclusive, furthermore, regeneration and repair of cartilage after debridement is also a great challenge for the treatment of OLT. Platelet-rich plasma (PRP) with good repair effect on cartilage injury is gradually applied in the treatment of OLT. However, there still lacks the unified understanding of the technique and specification of PRP for the treatment of OLT. Therefore, National Orthopedics Center of Shanghai Sixth People′s Hospital allied Foot Ankle Basic Research & Orthopedics Group, Chinese Association of Orthopedic Surgeons; Foot and Ankle Committee of Chinese Association of Sports Medicine Physicians; and Foot and Ankle Group of Orthopedic Specialized Branch of Shanghai Medical Association to organize related experts to formulate the Expert consensus on platelet- rich plasma treatment for osteochondral lesion of talus ( version2023). Fifteen recommendations were put forward upon PRP preparation, indications, contraindications and treatment methods of PRP for OLT, so as to standardize the PRP treatment for OLT.

Objective To study the expression of serum interleukin(IL)-17A,calcium binding protein S100A8 and S100A9 in children with severe Mycoplasma pneumoniae pneumonia(SMPP)and their prognostic significance.Methods A total of 116 children with SMPP who were diagnosed and treated in this hospital from March 2019 to March 2021 were enrolled as the SMPP group.According to the pediatric critical cases score,the SMPP children divided into non-critical group(43 cases),critial group(40 cases),extremely critical group(33 cases).According to the prognosis of 28 d after admission,the SMPP children were divided into a good prognosis group with 82 children and a poor prognosis group with 34 children.A total of 60 physical ex-amination of healthy children in the same hospital during the same period were enrolled as the control group.The levels of serum IL-17A,S100A8,S100A9,procalcitonin(PCT),C-reactive protein(CRP),IL-6 and tumor necrosis factor(TNF)-α were detected in each group.Pearson correlation analysis was used to analyze the cor-relation between serum levels of IL-17A,S100A8,S100A9 and PCT,CRP,IL-6,and TNF-α.Multivariate Lo-gistic regression analysis was used to analyze the factors affecting the poor prognosis of children with SMPP.The receiver operating characteristic(ROC)curve was used to analyze the value of each index in predicting the poor prognosis of children with SMPP.Results The SMPP group had significantly higher serum levels of IL-17A,S100A8,S100A9,PCT,CRP,IL-6,and TNF-α than the control group(P<0.05).In children with SMPP,the serum levels of IL-17A,S100A8,and S100A9 were positively correlated with PCT,CRP,IL-6,and TNF-α(P<0.05).The serum levels of IL-17A,S100A8 and S100A9 in extremely critical group were signifi-cantly higher than those in critical group and non-critical group(P<0.05).Elevated serum levels of IL-17A,S100A8 and S100A9 were independent risk factors for poor prognosis in children with SMPP.The area under the curve(AUC)of combined detection of serum IL-17A,S100A8 and S100A9 for predicting poor prognosis in children with SMPP was 0.895,which was higher than that of single detection of serum IL-17A,S100A8 and S100A9(0.833,0.764,0.810),the differences were all statistically significant(Z=3.780,6.723,5.012,P<0.059).The sensitivity and specificity of combined detection were 0.891 and 0.755,respectively.Conclu-sion The serum levels of IL-17A,S100A8 and S100A9 are increased in children with SMPP,which are related to the severity of SMPP.The combined detection of the three indicators has a high predictive value for the poor prognosis of SMPP.

Since outbreak of the coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)at the end of 2019,hundreds of millions of people were infected and millions of people around the world were killed.SARS-CoV-2 is highly prone to present rapid mutations that can render vaccines ineffective.Drug therapy is currently an effective mean to prevent and treat COVID-19,and traditional Chinese medicine therapy based on clinical char-acteristics and syndrome differentiation has played an extremely important role in the prevention and treatment of COVID-19 in China.Therefore,the characteristics and mechanism of the various prescrip-tions via multiple approaches and targets used in the treatment of COVID-19,including drugs with func-tions of heat-clearing,exterior-resolving,aromatic flavor and damp-resolving,and deficiency-tonifying,based on the clinical experience of Chinese traditional medicine in preventing and treating COVID-19,in order to provide reference for the clinical treatment of such diseases with symptomatic prescriptions.

Since outbreak of the coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)at the end of 2019,hundreds of millions of people were infected and millions of people around the world were killed.SARS-CoV-2 is highly prone to present rapid mutations that can render vaccines ineffective.Drug therapy is currently an effective mean to prevent and treat COVID-19,and traditional Chinese medicine therapy based on clinical char-acteristics and syndrome differentiation has played an extremely important role in the prevention and treatment of COVID-19 in China.Therefore,the characteristics and mechanism of the various prescrip-tions via multiple approaches and targets used in the treatment of COVID-19,including drugs with func-tions of heat-clearing,exterior-resolving,aromatic flavor and damp-resolving,and deficiency-tonifying,based on the clinical experience of Chinese traditional medicine in preventing and treating COVID-19,in order to provide reference for the clinical treatment of such diseases with symptomatic prescriptions.

Wilson's disease (WD) is a rare autosomal recessive disorder with a complex pathogenesis involving multiple systems, multiple visceral organs, and the complex copper homeostasis regulation system within the body. The liver is the most common organ for copper deposition, and liver injury is the earliest and most common manifestation of WD; therefore, it is important to find an ideal animal model for WD research. By summarizing the animal models of WD commonly used in the world, this article systematically summarizes the background, liver and nervous manifestations, and application of different models and compares the characteristics of different animal models, so as to provide a reference for the application of various animal models of WD.

AIM: To explore the promoting effect of 2-APB on skin wound healing in mice and its potential mechanism. METHODS: KM mice were divided into 5 groups: control group, DMSO group, low (50 mg/L), medium (100 mg/L) and high (200 mg/L) concentration 2-APB group. On the back of each mouse's skin use a circular punch about 1 cm on both sides of the midline of the spine to make a skin wound with a diameter of 10 mm and as deep as the fascia. The control group was only wrapped with gauze and no drugs were applied; the DMSO group was applied 1 g DMSO/Vaseline ointment per day; in the 2-APB group, apply 1 g of 2-APB/Vaseline ointment at a corresponding concentration every day. Pictures were taken the next day to observe the healing, and the material was taken on the 21st day, HE staining was used to observe the pathological morphology of the wound and western blot to detect TRPM7, TGF-β, collagen-I and IL-1β expression. RESULTS: Compared with the control group and the DMSO group, different concentrations of 2-APB could significantly promote skin wound healing in mice (P<0.01), but there was no significant difference in wound healing rate between the DMSO group and the control group group. The results of HE staining showed that, compared with the control group group and the DMSO group, 2-APB could increase the collagen content of the wound and the thickness of the dermis (P<0.01), but there was no significant difference between the DMSO group and the control group group. At the same time, 2-APB could also significantly increase the expression of TGF-β and Col-I on the wound, and inhibit the expression of TRPM7 and IL-1β. CONCLUSION: Different concentrations of 2-APB (50, 100 and 200 mg/L) can promote skin wound healing, and its mechanism may be related to the inhibition of TRPM7.