BACKGROUND:Steroid-induced femoral head necrosis is mostly caused by long-term and extensive use of hormones,but its specific pathogenesis is not yet clear and needs further study. OBJECTIVE:To screen out the differential miRNAs in osteoclast exosomes after the intervention of Panax notoginseng saponins,and on this basis,to further construct an osteogenic-related ferroptosis regulatory network to explore the potential mechanism and research direction of steroid-induced osteonecrosis of the femoral head. METHODS:MTT assay was used to detect the toxic effects of different concentrations of dexamethasone and different mass concentrations of Panax notoginseng saponins on Raw264.7 cell line.Tartrate resistant acid phosphatase staining and TUNEL assay were used to detect the effects of Panax notoginseng saponins on osteoclast inhibition and apoptosis.Exosomes were extracted from cultured osteoclasts with Panax notoginseng saponins intervention.Exosomes from different groups were sequenced to identify differentially expressed miRNAs.CytoScape 3.9.1 was used to construct and visualize the regulatory network between differentially expressed miRNAs and mRNAs.Candidate mRNAs were screened by GO analysis and KEGG analysis.Finally,the differential genes related to ferroptosis were screened out,and the regulatory network of ferroptosis-related genes was constructed. RESULTS AND CONCLUSION:(1)The concentration of dexamethasone(0.1 μmol/L)and Panax notoginseng saponins(1 736.85 μg/mL)suitable for intervention of Raw264.7 cells was determined by MTT assay.(2)Panax notoginseng saponins had an inhibitory effect on osteoclasts and could promote their apoptosis.(3)Totally 20 differentially expressed miRNAs were identified from osteoclast-derived exosome samples,and 11 differentially expressed miRNAs related to osteogenesis were predicted by target mRNAs.The regulatory networks of 4 up-regulated differentially expressed miRNAs corresponding to 155 down-regulated candidate mRNAs and 7 down-regulated differentially expressed miRNAs corresponding to 238 up-regulated candidate mRNAs were constructed.(4)Twenty-four genes related to ferroptosis were screened out from the differential genes.Finally,12 networks were constructed(miR-98-5p/PTGS2,miR-23b-3p/PTGS2,miR-425-5p/TFRC,miR-133a-3p/TFRC,miR-185-5p/TFRC,miR-23b-3p/NFE2L2,miR-23b-3p/LAMP2,miR-98-5p/LAMP2,miR-182-5p/LAMP2,miR-182-5p/TLR4,miR-23b-3p/ZFP36,and miR-182-5p/ZFP36).These results indicate that Panax notoginseng saponins may regulate osteoblast ferroptosis by regulating the expression of miRNAs derived from osteoclast exosomes,thus providing a new idea for the study of the mechanism of steroid-induced femoral head necrosis.

Objective:To establish ultra-performance liquid chromatography (UPLC) fingerprint chromatograms for Erigeron breviscapu from different origins and storage conditions; To identify the Erigeron breviscapus from different habitats by chemometric analysis.Methods:Acquity UPLC HSS T3 (2.1 mm×100 mm,1.8 μm) chromatography column was used; mobile phase was 0.1% formic acid aqueous solution-acetonitrile; detection wavelength was 268 nm; flow rate was 0.25 ml/min; column temperature was 35 ℃; injection volume was 2 μl gradient elution. The UPLC fingerprint of Erigeron breviscapu from different origins was established. Similarity evaluation combined with chemometric analysis methods such as clustering analysis (CA), principal component analysis (PCA), orthogonal partial least squares method - discriminant analysis (OPLS-DA) were used to identify the origin of Breviscapine from different places. Fingerprint technology was used to evaluate the similarity of storage conditions for Erigeron breviscapu.Results:The UPLC fingerprint method met the methodological requirements. 30 batches of Erigeron breviscapus had 24 common peaks, and the similarity between Xingyi in Guizhou and Huize in Qujing was 0.702-0.783, while the similarity between Honghe Luli, Kunming Fuming and Dali was 0.861-0.970. All samples were divided into two categories according to their origin by CA: category Ⅰ: Xingyi in Guizhou and Huize in Qujing, and category Ⅱ: Dali, Kunming Fuming and Honghe Luli. The results of PCA were consistent with CA. OPLS-DA screened out 10 differential markers of Erigeron breviscapus from different habitats. Moreover, a total of 40 common peaks were identified in six batches of Erigeron breviscapus samples stored under different conditions. Based on the similarity analysis, Erigeron breviscapus samples stored at 30% humidity and 70% humidity were classified into two separate categories.Conclusions:The fingerprint method constructed in this study is stable and reliable, and the predictive ability and reliability of the OPLS-DA model are excellent. By combining the two methods, a clear division can be made between Erigeron breviscapus from Yunnan and Guizhou. Humidity conditions are an important factor in storing Erigeron breviscapus.

BACKGROUND:It has been hypothesized that PANoptosis may be involved in the pathologic process of osteoporosis,but there have been no studies addressing the mechanisms of PANoptosis genes in osteoporosis. OBJECTIVE:To analyze the biological mechanism of PANoptosis regulators in the occurrence and development of osteoporosis. METHODS:The GSE56815 dataset was obtained from the Gene Expression Omnibus database and PANoptosis genes were extracted for differential analysis.The key genes of PANoptosis were screened by random forest tree model to construct a disease risk prediction model.Consensus clustering algorithm,single sample genome enrichment analysis and immune infiltration analysis were used to explore the differences between different PANoptosis molecular subtypes.Herbal drugs that regulate the key genes of PANoptosis were predicted through Coremine medical database,a medical ontology information retrieval platform. RESULTS AND CONCLUSION:Based on the four PANoptosis key genes(CASP1,CASP10,MEFV,and TNF),the diagnostic markers of osteoporosis were determined,and the risk prediction model was constructed and verified.Osteoporosis was divided into two different PANoptosis subtypes(clusters A,B and gene clusters A,B),and the PANoptosis scores of cluster B and gene cluster B were higher than those of cluster A and gene cluster A,respectively.Traditional Chinese drugs such as ginseng which can regulate the key genes of PANoptosis were predicted by the Coremine medical database.

Objective To investigate the regulation of insulin VGLUT2 gene expression in pancreatic β cells and its exactly mechanism.Methods The rat pancreatic β cell line RIN-5F was treated by different insulin concentrations and different siRNA.The changes of VGLUT2 mRNA and protein expression levels were detected by adopting real-time qPCR and Western blot.Results In-sulin with a concentration of 100,200 nmol/L significantly inhibited the expressions of VGLUT2 mRNA and protein in RIN-5RF cells(P<0.05),moreover the inhibiting effect was most significant at 100 nmol/L.After 100 nmol/L insulin treatment,the expres-sions of VGLUT2 mRNA and protein at 12,18,24 h were significantly inhibited compared with that at 0 h(P<0.05).Compared with the Blank group,Lip2000 group and Control-siRNA group,after interfering RIN-5F by using IR-siRNA,IRS1-siRNA and IRS2-siRNA,the inhibition situation of VGLUT2 mRNA and protein expressions by 100 nmol/L insulin in each group was signifi-cantly recovered(P<0.05).Conclusion Insulin at low concentration could inhibit VGLUT 2 gene expression in rat pancreatic β cell line RIN-5F.

Objective To investigate the expression of Akt and phosphoryled Akt (p-Akt1) protein in pancreatic carcinoma and to determine the clinical significance. Methods In 74 cases of pancreatic carcinoma and 10 cases of normal pancreatic tissue samples, the expression of Akt and p-Akt1 were detected by immunohistochemical method, and the its relationship with clinicopathologic characteristics and prognosis were analyzed. Results The positive expression rate of Akt and p-Akt1 in pancreatic carcinoma were 87.8% and 83.8% respectively, while there was no expression of Akt and p-Akt1 in normal pancreatic tissue, and the difference was statistically significant (p < 0.05). There was a positive correlation between the expression of Akt and p-Akt1 in pancreatic carcinoma (r =0.274, P =0. 018). The expression of Akt and p-Akt1 was not significantly associated with the age, sex, location, size, pathology stages, lymph nodes metastasis, clinical stages and nerve invasion of the tumor (P >0.05). But the higher expression of p-Akt1 was associated with T stages and TNM staging (p =0. 002). Patients with high intensity of Akt and p-Akt1 expression showed a significantly longer median survival time [(16.0 ± 5.7) month and (23.0 ± 5.5) month, respectively]than those with low intensity expression [(9.3 ± 0.2) month and (11.1 ± 1.8) month (P = 0. 007 and P = 0.004) respectively]. Conclusions p-Akt1 expression is a significant positive prognostic factor for pancreaticcarcinoma and detection of p-Akt1 expression may be of clinical value.