Clinical Effect of Modified Gegen Qinlian Decoction Combined with Western Medicine on Children with Rotavirus Enteritis with Damp-Heat Syndrome

[Objectives] To explore the clinical effect of Modified Gegen Qinlian Decoction combined with western medicine on children with rotavirus enteritis with damp-heat syndrome and its influence on myocardial enzymes.

Celastrol targeting Nedd4 reduces Nrf2-mediated oxidative stress in astrocytes after ischemic stroke

Stroke is the second leading cause of death worldwide,and oxidative stress plays a crucial role.Celastrol exhibits strong antioxidant properties in several diseases;however,whether it can affect oxidation in cerebral ischemic-reperfusion injury(CIRI)remains unclear.This study aimed to determine whether celastrol could reduce oxidative damage during CIRI and to elucidate the underlying mechanisms.Here,we found that celastrol attenuated oxidative injury in CIRI by upregulating nuclear factor E2-related factor 2(Nrf2).Using alkynyl-tagged celastrol and liquid chromatography-tandem mass spectrometry,we showed that celastrol directly bound to neuronally expressed developmentally downregulated 4(Nedd4)and then released Nrf2 from Nedd4 in astrocytes.Nedd4 promoted the degradation of Nrf2 through K48-linked ubiquitination and thus contributed to astrocytic reactive oxygen species production in CIRI,which was significantly blocked by celastrol.Furthermore,by inhibiting oxidative stress and astrocyte activation,celastrol effectively rescued neurons from axon damage and apoptosis.Our study uncovered Nedd4 as a direct target of celastrol,and that celastrol exerts an antioxidative effect on astrocytes by inhibiting the interaction between Nedd4 and Nrf2 and reducing Nrf2 degradation in CIRI.

Monoclonal antibody targeting mu-opioid receptor attenuates morphine tolerance via enhancing morphine-induced receptor endocytosis

Morphine is a frequently used analgesic that activates the mu-opioid receptor(MOR),which has prominent side effects of tolerance.Although the inefficiency of morphine in inducing the endocytosis of MOR underlies the development of morphine tolerance,currently,there is no effective therapy to treat morphine tolerance.In the current study,we aimed to develop a monoclonal antibody(mAb)precisely targeting MOR and to determine its therapeutic efficacy on morphine tolerance and the underlying molecular mechanisms.We successfully prepared a mAb targeting MOR,named 3A5C7,by hybridoma technique using a strategy of deoxyribonucleic acid immunization combined with cell immunization,and identified it as an immunoglobulin G mAb with high specificity and affinity for MOR and binding ability to antigens with spatial conformation.Treatment of two cell lines,HEK293T and SH-SY5Y,with 3A5C7 enhanced morphine-induced MOR endocytosis via a G protein-coupled receptor kinase 2(GRK2)/b-arrestin2-dependent mechanism,as demonstrated by immunofluorescence staining,flow cytometry,Western blotting,coimmunoprecipitation,and small interfering ribonucleic acid(siRNA)-based knockdown.This mAb also allowed MOR recycling from cytoplasm to plasma membrane and attenuated morphine-induced phosphorylation of MOR.We established an in vitro morphine tolerance model using differentiated SH-SY5Y cells induced by retinoic acid.Western blot,enzyme-linked immunosorbent assays,and siRNA-based knockdown revealed that 3A5C7 mAb diminished hyperactivation of adenylate cyclase,the in vitro biomarker of morphine tolerance,via the GRK2/b-arrestin2 pathway.Furthermore,in vivo hotplate test demonstrated that chronic intrathecal administration of 3A5C7 significantly alleviated morphine tolerance in mice,and withdrawal jumping test revealed that both chronic and acute 3A5C7 intrathecal administration attenuated morphine dependence.Finally,intrathecal electroporation of silencing short hairpin RNA illustrated that the in vivo anti-tolerance and anti-dependence efficacy of 3A5C7 was mediated by enhanced morphine-induced MOR endocytosis via GRK2/b-arrestin2 pathway.Collectively,our study provided a therapeutic mAb,3A5C7,targeting MOR to treat morphine tolerance,mediated by enhancing morphine-induced MOR endocytosis.The mAb 3A5C7 demonstrates promising translational value to treat clinical morphine tolerance.

MicroRNAs as prognostic biomarkers for survival outcome in osteosarcoma:A meta-analysis

BACKGROUND Osteosarcoma was considered to be one of the most prevalent malignant bone tumors in adolescents.AIM To explore the prognostic significance of microRNA(miRNA)in osteosarcoma.METHODS The literature was selected by searching online in PubMed,Embase,Web of Science,Cochrane Library,China National Knowledge Infrastructure,and Wanfang Database until July 1,2021.The pooled hazard ratio(HR)with corresponding 95%confidence interval(CI)for the outcomes of overall survival(OS),disease-free survival(DFS),progression-free survival(PFS)and recurrence-free survival were calculated.Subgroup analyses were carried out to identify potential sources of heterogeneity.Publication bias was assessed by Egger’s bias indicator test.RESULTS A total of 60 studies from 54 articles with 5824 osteosarcoma patients were included for this meta-analysis.The pooled HR for OS,DFS,PFS were 2.92(95%CI:2.43-3.41,P=0.000),3.70(95%CI:2.80-4.61,P=0.000),and 3.57(95%CI:1.60-5.54,P=0.000),respectively.The high miR-21 expression levels were related to poor OS in osteosarcoma(HR=2.86,95%CI:1.20-4.53,P=0.001).Subgroup analysis demonstrated that a high expression level of miRNA correlated with worse OS(HR:3.56,95%CI:2.59-4.54,P=0.000).In addition,miRNA from tissue(HR:3.20,95%CI:2.16-4.23,P=0.000)may be a stronger prognostic biomarker in comparison with that from serum and plasma.CONCLUSION miRNA(especially miR-21)could be served as a potential prognostic biomarker for osteosarcoma.A high expression level of miRNA in tumor tissue correlated with worse OS of osteosarcoma.