OBJECTIVES: To investigate the regulatory role of the NLRP3 signaling pathway in hepatocyte pyroptosis in nonalcoholic steatohepatitis (NASH) under hypoxia. METHODS: Twenty-four male C57BL/6 mice were randomized equally into hypoxic control (A), hypoxic NASH model (B), hypoxic NASH+NLRP3 inhibitor (C), and hypoxic NASH+caspase-1 inhibitor (D) groups. In groups B-D, the mice were fed a methionine choline-deficient (MCD) diet under hypoxic conditions (to simulate a 5000 m altitude) for 6 weeks; the mice in groups C and D received intraperitoneal injections of the respective inhibitors every other day. RESULTS: Compared with those in group A, the mice in group B showed significantly elevated serum levels of FBG, TC, TG, ALT and AST, increased liver lipid content, inflammatory cell infiltration and collagen fiber deposition, and enhanced hepatic expressions of NLRP3, caspase-1, IL-1β and GSDMD proteins, with obvious swelling, cristae breakage, vacuolization, and outer membrane disruption of the mitochondria, ribosome loss in the cytoplasm, destruction of the nuclear membrane, and pathological changes of the rough endoplasmic reticulum. Treatment with NLRP3 inhibitor and caspase-1 inhibitor both significantly lowered serum levels of TC, TG, ALT and AST (but without significantly affecting FBG) in the mouse models, and reduced liver lipid content, inflammatory cell infiltration, collagen deposition, and expression levels of NLRP3, caspase-1, GSDMD and IL-1β. The treatments also significantly improved pathological changes in the mitochondria, ribosomes and endoplasmic reticulum in liver tissues of the mice. CONCLUSIONS NLRP3 signaling pathway plays a key role in promoting hepatocyte pyroptosis in NASH mice under hypoxic condition, and inhibiting this pathway can effectively reduce liver inflammation, suggesting its potential as a therapeutic target for NASH treatment.
Animals ; Non-alcoholic Fatty Liver Disease/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Pyroptosis ; Mice, Inbred C57BL ; Male ; Hepatocytes/pathology* ; Signal Transduction ; Mice ; Hypoxia/metabolism* ; Caspase 1/metabolism* ; Interleukin-1beta/metabolism* ; Liver/metabolism*

Objective To establish the UPLC fingerprint of 10 batches of Gongliuning capsule and the content determination methods of 10 main components;To conduct network pharmacological research;To provide scientific basis for predicting the quality marker of Gongliuning capsule.Methods UPLC-DAD technique was used to establish the fingerprint of different batches of Gongliuning capsule samples and obtain the complete chemical composition information of Gongliuning capsule.With the help of network pharmacological research techniques,the target of chemical components of Gongliuning capsule and related pathways screened from various databases were predicted and analyzed,and the protein interaction network and composition-target-pathway diagram were constructed to predict the effective components of Gongliuning capsule.Based on the obtained pharmacodynamic substances,the content determination method of Gongliuning capsule was established to evaluate the quality of Gongliuning capsule.Results The fingerprints of Gongliuning capsule identified 17 common peaks.Combined with network pharmacology method,the candidate chemical components were analyzed,10 core targets and 42 metabolic pathways were obtained.Based on the testability of components and the availability of reference substances,10 compounds with high contribution were selected for content determination.Methodological verification results showed that all the 10 compounds had good linear relationship(r2>0.999 4)in the detection range,with good precision,accuracy and stability.Conclusion The established UPLC fingerprint,network pharmacology and multiple component content determination methods are highly specific,accurate and reliable.Combined with chemical pattern recognition,they can be effectively used for quality control and quality marker prediction of Gongliuning capsules.

AIM To investigate the protective effects of the aqueous extract of Bulbophyllum kwangtungense Schltr.on D-galactose(D-gal),an oxidative stress-induced liver damage in rats.METHODS 60 Male SD rats were randomly divided into the normal control group,the model group,the resveratrol group(20 mg/kg),and low-dose,medium-dose,and high-dose B.kwangtungense aqueous extract groups(750,1 500 and 3 000 mg/kg).Except for those of the normal control group,the rats of the other groups were subcutaneously injected with D-gal(200 mg/kg)in the back of neck to establish a liver damage model.Simultaneously,each group underwent the corresponding drug administration by gavage(10 mL/kg)once daily for 42 days.After the treatment period,the rats had their peripheral blood and liver tissues collected for the calculation of the liver index;their serum AST and ALT activities,hepatic SOD and GSH-Px activities,and MDA levels measured;their hepatic pathological changes observed using HE staining;their hepatic mRNA expressions of Nrf2,NQO-1 and HO-1 detected by RT-qPCR;and their hepatic protein expressions of Nrf2,NQO-1,HO-1,Bax,Bcl-2 and cleaved Caspase-3 measured by Western blot.RESULTS Compared with the model group,the groups intervened with medium-dose or high-dose B.kwangtungense aqueous extract showed increased liver index level(P＜0.01);decreased serum AST and ALT activities(P＜0.01);increased hepatic SOD and GSH-Px activities(P＜0.05,P＜0.01);reduced MDA levels(P＜0.01);alleviated pathological liver damage and increased hepatic mRNA and protein expressions of Nrf2,NQO-1 and HO-1(P＜0.05,P＜0.01);and reduced protein expressions of cleaved Caspase-3 and the Bax/Bcl-2 ratio as well(P＜0.05,P＜0.01).CONCLUSION The aqueous extract of B.kwangtungense is protective of rats with D-gal-induced liver injury,and the underlying mechanism may associate with the inhibition of oxidative stress and the suppression of hepatocellular apoptosis mediated by the activation of the Nrf2/HO-1/NQO-1 signaling pathway.

AIM To study the protective effect of Gynura divaricate polysaccharide on gouty nephropathy(GN)induced by dry yeast combined with adenine in rats.METHODS Sixty male SD rats were randomly divided into the normal group,the model group,the allopurinol group(42 mg/kg),and the low-dose,medium-dose and high-dose G.divaricate polysaccharide groups(140,280,560 mg/kg).All the rats except those of the normal group were induced into GN models by intragastrical dosing of yeast(5 g/kg)and adenine(100 mg/kg)and intervened with corresponding drug administration simultaneously.After 35 days,the rats had their levels of creatinine(Cr)and uric acid(UA)in serum and urine detected and their fraction excretion of uric acid(FEUA)value determined;their kidney mass and volume measured and their levels of kidney index and density calculated;their renal pathological changes checked by HE staining;their renal GLUT9,URAT1,ABCG2 and OAT1 mRNA expressions dectected by RT-qPCR;and their renal GLUT9,URAT1,ABCG2 and OAT1 protein expressions dectected by Western blot.RESULTS Compared with the model group,each dose of G.divaricate polysaccharide group displayed decreased levels of kidney mass,kidney volume and kidney index(P＜0.01);increased levels of kidney density(P＜0.05,P＜0.01);decreased serum levels of UA and Cr(P＜0.01);increased urine levels of UA and Cr(P＜0.01);increased FEUA value(P＜0.01);decreased GLUT9,URAT1 mRNA and protein expressions(P＜0.05,P＜0.01);and increased ABCG2,OAT1 mRNA and protein expressions(P＜0.05,P＜0.01);and more alleviated renal histological aberrations.CONCLUSION G.divaricate polysaccharide exerts good protective effects against yeast/adenine-induced GN in rats probably through down-regulating protein expression of GLUT9,URAT1 and up-regulating ABCG2 and OAT1.

Objective To analyze the current quality of treatment for hospitalized cancer patients in Bei-jing,identify major issues in treatment practices,and propose improvements.Methods Nine hospitals in Beijing were selected for examination.Expert on-site interviews and medical record sampling were conducted.The"Bei-jing Cancer Diagnosis and Treatment Quality Control Checklist"was used to assess the hardware,management,anti-cancer drug therapy,radiation therapy,and surgical treatment during cancer treatment at these hospitals from January to October 2023.The relevant problems were analyzed.Results Among the nine hospitals,two(22.2％)were equipped with laminar flow rooms,and three(33.3％)had intravenous drug preparation centers.In terms of institutional management,seven hospitals(77.8％)had standardized anti-cancer drug prescription authority management,eight(88.9％)had complete emergency plans,and five(55.6％)had oncology specialist pharmacists.Regarding anti-cancer drug therapy,the areas with higher completion rates included pathology diag-nosis support(97.6％),routine pre-treatment examinations(96.3％),adverse reaction evaluation(92.7％),discharge summaries(95.1％),and admission records(91.5％).However,the accuracy of tumor staging before treatment(70.7％)and the evaluation of therapeutic efficacy after drug treatment(76.9％)needed improvement.The oncology specialty significantly outperformed the non-oncology specialty in terms of the accuracy rate of TNM staging(86.0％vs.46.9％,P＜0.001),the completeness of informed consent forms(100％vs.68.8％,P＜0.001),the completeness of drug indication evaluation(96.0％vs.78.1％,P=0.025),the completeness of admission medical history records(98.0％vs.81.3％,P=0.008),the rationality of drug dosage(96.0％vs.75.0％,P=0.005),the rationality of drug infusion time(100％vs.62.5％,P＜0.001),and the rationality of the order of drug infusion(100％vs.87.5％,P=0.010).Although the quality of radiation therapy was high,the subsequent evaluation of therapeutic efficacy(39.3％)requires enhancement.In surgical treatment,the preoper-ative pathology diagnosis support rate(78.1％)and the accuracy of tumor staging(37.5％)were relatively low,indicating issues with incomplete preoperative evaluation and the absence of multidisciplinary discussions.Conclusions There remains significant room for improvement in the quality of cancer treatment in China.It is recommended to standardize tumor staging assessment processes,strengthen entry assessments for non-oncology departments,promote the implementation of multidisciplinary treatment models,and establish a multi-department collaborative management model.Continuous monitoring of cancer diagnosis and treatment quality indicators is es-sential to promote ongoing improvements in cancer treatment quality.

Objective To investigate the predictive value of serum total IgE,eosinophil(EOS)ratio,lactate dehydrogenase(LDH)and exhaled nitric oxide(FeNO)levels in allergic rhinitis(AR)complicated with bronchial asthma(BA).Methods A total of 127 children with AR were divided into the AR group(84 cases)and the AR+BA group(43 cases)based on the presence of BA.Another 55 healthy children were selected as the control group.General data,total IgE,EOS ratio,LDH and FeNO levels were compared between the three groups.The correlation between these indicators and duration of disease symptoms,nasal symptom scores were analyzed.Receiver operating characteristics(ROC)curve analysis was performed to evaluate the predictive value of these indicators for AR and AR+BA.Results Total IgE,EOS ratio,LDH and FeNO levels were significantly higher in the AR group and the AR+BA group than those in the control group(P＜0.05).The total IgE,EOS ratio and FeNO levels were higher in the AR+BA group than those in the AR group(P＜0.05).Correlation analysis showed that total IgE,EOS ratio and FeNO levels were positively correlated with the duration of disease symptoms and symptom scores in the AR group and the AR+BA group(P＜0.05).In the AR group,LDH levels were positively correlated with the duration of disease symptoms and symptom scores(P＜0.05),but in the AR+BA group,there was no significant correlation between the duration of BA symptoms and symptom scores(P＞0.05).ROC curve analysis indicated that total IgE,FeNO,EOS ratio and LDH all had predictive values for both AR and AR+BA.The predictive value of total IgE for AR and AR+BA was higher(AUC=0.971 and 0.800).Combined detection of total IgE,EOS ratio,LDH and FeNO had the best predictive value for AR+BA(AUC=0.904).Conclusion Serum levels of total IgE,LDH,EOS ratio and FeNO can be served as effective indicators for predicting AR complicated with BA.Combined detection can improve the accuracy of prediction and provide a reference for early clinical identification and intervention in AR+BA.

Objective To investigate the effect of a nucleic acid purifier combined with a vacuum concentrator on DNA recovery rate.Methods Using 39 tubes of 007 standard samples,two 30 μL samples were taken from each tube.Samples were purified using the PrepFiler ExpressTM BTA Purification Kit on the AutoMate ExpressTM purifier,with elution volumes of 30 μL and 200 μL.Samples with 200 μL eluent were concentrated to 30 μL using a trace DNA sample vacuum concentrator.DNA solution concentrations were measured,and recovery rates were calculated and compared.Results The measured DNA concentration of the 007 standard samples was 0.08～0.15(0.11±0.02)ng/μL,showing a significant difference from the kit-labeled concentration of 0.1 ng/μL(t=3.88,P<0.01).The DNA solution concentration in the purified and concentrated group(200 μL eluent concentrated to 30 μL)[0.05～0.13(0.08±0.02)ng/μL]was significantly higher than the purified group(30 μL eluent)[0.02～0.08(0.05±0.01)ng/μL](t=21.20,P<0.01).The DNA recovery rate of the purified and concentrated group[54.55%～92.86%(75.41±10.12)%]was substantially higher than the purified group[25.00%～54.55%(39.79±7.13)%](t=19.79,P<0.01).Conclusion Utilizing a large-volume elution system in nucleic acid purification can significantly enhance DNA recovery rate.Combining this method with a vacuum concentrator effectively increases DNA solution concentration,thereby improving DNA testing success rates.

Objective:To explore the application and clinical significance of pathological diagnostic criteria for medically refractory epilepsy in children.Methods:Cross-sectional study.A retrospective analysis was conducted on 490 children(pathology involved) with medically refractory epilepsy treated continuously in the Pediatric Epilepsy Center of Peking University First Hospital from January 2019 to May 2022.The distribution of different pathological types was observed, and the differences in clinical characteristics among different pathological types were analyzed through Kruskal-Wallis or χ2 tests.The impact of clinical and pathological features on patient prognosis was evaluated through regression analysis. Results:Focal cortical dysplasia (FCD) was the predominant lesion (49.59%, 243/490).The electroencephalograms ( χ2=6.720, P=0.035) and clinical seizure characteristics ( χ2=26.370, P<0.001) in FCDⅡ were more focal than those in FCD Ⅰ and Ⅲ.Moreover, the proportions of focal resection in surgery ( χ2=24.286, P<0.001) and central involvement ( χ2=22.849, P<0.001) in FCDⅡ were higher than those in FCD Ⅰ and Ⅲ.Univariate and multivariate regression analyses revealed that FCD Ⅱ had a better prognosis than other dysplastic patients among the 375 cases of dysplasia ( P=0.049).Next-generation sequencing was performed on 35 cases of cortical malformations with such morphological characteristics as increased numbers of neurons in the white matter and Olig2-positive glial cell hyperplasia, and SLC35A2 mutations were detected in 2 cases (5.71%). Conclusions:Pathology of refractory epilepsy is specialized and continuously evolving.Standardized specimen processing and the accumulation of morphological, immunohistochemical, and molecular genetic data provide the foundation for clarifying the neuropathological nature of epilepsy, improving integrated classification, and advancing prognosis prediction and targeted therapy.

Objective To investigate the predictive value of serum total IgE,eosinophil(EOS)ratio,lactate dehydrogenase(LDH)and exhaled nitric oxide(FeNO)levels in allergic rhinitis(AR)complicated with bronchial asthma(BA).Methods A total of 127 children with AR were divided into the AR group(84 cases)and the AR+BA group(43 cases)based on the presence of BA.Another 55 healthy children were selected as the control group.General data,total IgE,EOS ratio,LDH and FeNO levels were compared between the three groups.The correlation between these indicators and duration of disease symptoms,nasal symptom scores were analyzed.Receiver operating characteristics(ROC)curve analysis was performed to evaluate the predictive value of these indicators for AR and AR+BA.Results Total IgE,EOS ratio,LDH and FeNO levels were significantly higher in the AR group and the AR+BA group than those in the control group(P＜0.05).The total IgE,EOS ratio and FeNO levels were higher in the AR+BA group than those in the AR group(P＜0.05).Correlation analysis showed that total IgE,EOS ratio and FeNO levels were positively correlated with the duration of disease symptoms and symptom scores in the AR group and the AR+BA group(P＜0.05).In the AR group,LDH levels were positively correlated with the duration of disease symptoms and symptom scores(P＜0.05),but in the AR+BA group,there was no significant correlation between the duration of BA symptoms and symptom scores(P＞0.05).ROC curve analysis indicated that total IgE,FeNO,EOS ratio and LDH all had predictive values for both AR and AR+BA.The predictive value of total IgE for AR and AR+BA was higher(AUC=0.971 and 0.800).Combined detection of total IgE,EOS ratio,LDH and FeNO had the best predictive value for AR+BA(AUC=0.904).Conclusion Serum levels of total IgE,LDH,EOS ratio and FeNO can be served as effective indicators for predicting AR complicated with BA.Combined detection can improve the accuracy of prediction and provide a reference for early clinical identification and intervention in AR+BA.

AIM To investigate the protective effects of the aqueous extract of Bulbophyllum kwangtungense Schltr.on D-galactose(D-gal),an oxidative stress-induced liver damage in rats.METHODS 60 Male SD rats were randomly divided into the normal control group,the model group,the resveratrol group(20 mg/kg),and low-dose,medium-dose,and high-dose B.kwangtungense aqueous extract groups(750,1 500 and 3 000 mg/kg).Except for those of the normal control group,the rats of the other groups were subcutaneously injected with D-gal(200 mg/kg)in the back of neck to establish a liver damage model.Simultaneously,each group underwent the corresponding drug administration by gavage(10 mL/kg)once daily for 42 days.After the treatment period,the rats had their peripheral blood and liver tissues collected for the calculation of the liver index;their serum AST and ALT activities,hepatic SOD and GSH-Px activities,and MDA levels measured;their hepatic pathological changes observed using HE staining;their hepatic mRNA expressions of Nrf2,NQO-1 and HO-1 detected by RT-qPCR;and their hepatic protein expressions of Nrf2,NQO-1,HO-1,Bax,Bcl-2 and cleaved Caspase-3 measured by Western blot.RESULTS Compared with the model group,the groups intervened with medium-dose or high-dose B.kwangtungense aqueous extract showed increased liver index level(P＜0.01);decreased serum AST and ALT activities(P＜0.01);increased hepatic SOD and GSH-Px activities(P＜0.05,P＜0.01);reduced MDA levels(P＜0.01);alleviated pathological liver damage and increased hepatic mRNA and protein expressions of Nrf2,NQO-1 and HO-1(P＜0.05,P＜0.01);and reduced protein expressions of cleaved Caspase-3 and the Bax/Bcl-2 ratio as well(P＜0.05,P＜0.01).CONCLUSION The aqueous extract of B.kwangtungense is protective of rats with D-gal-induced liver injury,and the underlying mechanism may associate with the inhibition of oxidative stress and the suppression of hepatocellular apoptosis mediated by the activation of the Nrf2/HO-1/NQO-1 signaling pathway.