BACKGROUND: The transcription factor Runx2 is the key factor that regulates osteogenic differention and bone development. It has been reported that the C2C12 mesenchymal cells can be induced to differentiate into osteoblasts by Runx2 overexpression, but the molecular mechanism of induction is stil largely unclear. OBJECTIVE: To investigate the role of the members of the miR-376 family during Runx2-induced osteogenic differentiation in C2C12 cells. METHODS: The expression of the members of the miR-376 family was detected by real-time quantitative PCR at different time points using C2C12/Runx2Dox sub-line with conditional Runx2 expression. In miR-376b-3p-transfected C2C12/Runx2Dox cells, the expression of osteoblast markers, such as alkaline phosphatase and osteocalcin, was detected by real-time quantitative PCR, and the alkaline phosphatase activity was also examined by alkaline phosphatase staining. The putative miR-376b-3p targets were commonly predicted by online tools (miRanda, miRWalk and TargetScan). The functional classification of these putative targets was performed by DAVID Bioinformatics Resources database. RESULTS AND CONCLUSION: The expression of miR-376b-3p was significantly increased during Runx2-induced osteogenic differentiation of C2C12 cells, but the expression of other members was not changed. Transfection of miR-376b-3p mimic upregulated alkaline phosphatase expression, but had no effect on osteocalcin expression. The alkaline phosphatase activity was also increased by transfection of miR-376b-3p. The functional classification of miR-376b-3p putative targets showed that miR-376b-3p is involved in the skeleton development, indicating the role of miR-376b-3p in osteoblast differentiation. Taken together, these results suggest that Runx2 promotes early osteogenic differentiation in C2C12 cells by regulating the expression of genes related to osteogenic differentiation through upregulation of miR-376b-3p.

Aim To investigate TY501′s role in bleo-mycin ( BLM )-induced pulmonary fibrosis in rats. Methods Forty rats were randomly divided into 5 groups, including the sham operation, BLM, PFD, TY501(high and low dose) groups. After administra-tion of BLM intratracheally, PFD and TY501 were giv-en in each group daily, according to the dosage de-signed during 21 days. Lung coefficient, PaO2 were tested before killing the rats. The contents of ALB, ALP, LDH, GSH, HYP were detected by regent kit respectively. PCⅢ and COL4 were determined by ELISA. Results ( 1 ) Some indicators of alveolitis in early stage of IPF: the contents of lung coefficient in three treatment groups were lower and PaO2 was higher than those in BLM group ( P<0. 05 ); compared with BLM group, the contents of ALB, ALP, LDH in the treatment groups reduced on 21 st day ( P<0. 05 );the expression of GSH in BLM group was increased for feedback regulation and higher than the treatment groups and the sham operation group (P<0. 05);(2) some indicators of pulmonary interstitial fibrosis in late stage of IPF:the expressions of HYP, PCⅢand COL4 were reduced after the treatment. There were signifi-cant differences compared with BLM group ( P <0. 05 ) . Conclusions TY501 is valuable for the ther-apy of IPF, the same as the positive drug pirfenidone. TY501 attenuates BLM-induced pulmonary fibrosis, which may be related to the affection of TGF-βpathway and inhibition of MMPs.

ObjectiveTo explore the childhood trauma experience and its relation to the eye emotion recognition ability in patients with schizophrenia.MethodsSeventy-one schizophrenic patients and fifty-two healthy subjects were assessed with the Childhood Trauma Questionnaire (CTQ) and the Eye Basic Emotion Discrimination Task (EBEDT).ResultsThe CTQ total score in the patients group was significantly higher than those in the control group (47.86 ± 13.26 vs 34.27 ± 7.27,P ＜ 0.01 ).The subscores in emotional abuse,physical abuse,sexual abuse,emotional neglect and physical neglect of CTQ in the patients group were also significantly higher than those in the control group.(all P＜ 0.01 ).Compared with controls,the patients had significantly lower total accuracy( (86.74 ± 6.16vs73.42 ± 11.86) ％,P ＜ 0.01 ) and longer total reaction time ( (2776.75 ± 703.42 vs 4321.40 ± 1956.83 )ms,P ＜ 0.01 )of EBEDT.The significant difference were also found on the accuracy and reaction time of the EBEDT six basic emotion between the two groups ( all P ＜ 0.01 ).Correlation analysis indicated that CTQ total score were negatively correlated with the EBEDT total accuracy ( r=-0.390,P＜0.01 ),and its sub-accuracy of happiness,surprise,sadness,and anger ( respectively r =- 0.419,r =- 0.451,r =- 0.391,r =- 0.344,P ＜ 0.01 ) in the patients group.The correlation between CTQ total score and EBEDT total accuracy was also found in the control group.ConclusionChildhood trauma is related to eye basic emotion recognition in schizophrenia patients,suggesting that the trauma during childhood can affect the social recognition ability.

Adult ; Chromosome Aberrations ; Female ; Humans ; Male ; Pedigree ; Spinocerebellar Ataxias ; genetics

OBJECTIVETo isolate the side population (SP) and non-side population (NSP) cells from A431 cells and compare their difference in tumorigenicity in mice and the expression profiles of epithelial-mesenchymal transition (EMT) markers.

METHODSA431 cells stained with Hoechst 33342 were sorted with flow cytometry. The isolated SP cells and NSP cells were inoculated into NOD/SCID mice and the tumorigenicity of the cells was observed. EMT markers E-cadherin, β-catenin, vimentin, AXL, and Erbb3 in the tumor tissues were detected by immunohisto-staining.

RESULTSThe tumors generated by SP cells were larger than those by NSP cells in NOD/SCID mice. Compared with the tumors generated by NSP cells, the cells in the periphery of tumors generated by SP cells showed up-regulated expressions of AXL, vimentin and β-catenin and down-regulated ERBB3 and E-cadherin.

CONCLUSIONThe SP cells in A431 cells have a strong tumorigenicity and show more EMT phenotypes in tissues.

Animals ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Humans ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplastic Stem Cells ; metabolism ; Side-Population Cells ; metabolism ; Skin Neoplasms ; metabolism ; pathology

Objective:Enriching and isolating breast cancer stem cells from breast cancer transplantation tumors in nude mice.Methods:Human breast cancer MDA-MB-231 cells were injected into the right axilla subcutaneous of 20 nude mice, and the tumor growth was observed .After 30 days, tumors were isolated and stained with hematoxylin and eosin, and then tumor cells from tissues were isolated. DMEM medium containing serum was used to cultivate isolated transplantation tumor cells, cell morphology and growth were also observed. Flow cytometry was used to detect the proportion of stem cells (CD44 +/CD24 -/low cells) in transplantation tumor cells. Serum-free DMEM medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and B27 cell supplement were used to cultivate transplantation tumor cells and to obtain cell microspheres. The proportion of stem cells on the 10th day in cell microspheres was detected by using flow cell sorter and stem cells were isolated according to the markers of cell surface. Results:After subcutaneously injecting MDA-MB-231 cells into 20 nude mice for 9 days, 17 nude mice had subcutaneous tumors with more parenchymal cells, little interstitial cells, arranged cords tumor cells, large volume of the cell and abundant cytoplasm, the nuclei in different sizes and hyperchromatic state, mitotic more common, the nucleoli clear and obvious pleomorphy. After cultivating transplantation tumor cells with DMEM medium containing serum, the cells began to grow adherent after 24 h, and the adherent proportion rose to 60% after 3 days; after 7 days, the cell proliferation was accelerated; and the cell morphology was more consistent, most of which were spindle shaped and were not significantly different from MDA-MB-231 cells; the proportion of stem cells in transplantation tumor cells was (0.10±0.02)%. After cultivating transplantation tumor cells with serum-free DMEM medium containing cell cultured supplement, the cells grow in spherical patterns, the proportion of stem cells in cell microspheres got up to (70.47±2.03)% on the 10th day.Conclusions:Subcutaneously injecting MDA-MB-231 cells in nude mice can build breast cancer nude mice ectopic transplantation tumor model. Breast cancer stem cells in the transplantation tumors can be enriched from isolated transplantation tumor cells through serum-free medium, and more stem cells can be isolated to provide the research basis for the biological characteristics of breast cancer stem cells.

Objective To evaluate five types of DIC scoring systems based on sepsis patients,to explore the values of different DIC scoring systems in the occurrence and prognosis of DIC in sepsis patients,and to compare the applicability of different DIC scoring systems for sepsis complicated with DIC.Methods Laboratory indexes and clinical data from sepsis patients who had been hospitalized in Gansu Provincial People's Hospital from December 1,2019 to December 31,2021 were retrospectively analyzed within 24 hours.Five types of DIC scoring systems were used to score,and the difference of diagnostic rate and discharge outcome in sepsis patients with different severity was compared.The ROC curves of five DIC scoring systems were established to evaluate the accu-racy of DIC in sepsis patients.Results The fatality rate of sepsis increased with the severity of sepsis(P<0.05).There were statistically significant differences in discharge outcomes between DIC and non-DIC in the five scoring systems(P<0.05).JMHW,CDSS and part of ISTH were detected in JAAM cases,while ISTH was detected in non-dominant ISTH cases.ISTH,JAAM,JMHW,CDSS,and non-dominant ISTH5 scoring systems were used to diagnose DIC,and absence of full health restoration and death were 3.0,3.8,4.2,3.9,and 3.0 times higher than non-DIC cases,respectively.Conclusion JAAM scoring system has higher diagnostic rate and sensitivity for adult sepsis.CDSS and JMHW scoring systems are more accurate in predicting the prognosis of sepsis patients.

Objective The combined detection of serum angiopoietin-like protein 8(ANGPTL8)and Vascular cell adhesion molecule-1(VCAM-1)levels was analyzed for the predictive value of cerebral vasospasm(CVS)after intracranial aneurysm embolization.Methods A total of 196 patients who underwent intracranial aneurysm embolization in our hospital from March 2019-March 2022 were selected as the study subjects,99 patients with CVS were in the CVS group,and 97 patients without CVS were in the non CVS group.Serum ANGPTL8 and VCAM-1 levels were detected by ELISA;the correlation between serum ANGPTL8 and VCAM-1 levels was analyzed by Pearson method,Logistic regression was used to analyze the influencing factors of CVS in patients undergoing intracranial aneurysm embolization;ROC curve was used to analyze the serum levels of ANGPTL8 and VCAM-1 to predict the cutoff value of CVS in patients undergoing intracranial aneurysm embolization;four grid table method was used to analyze the predictive value of ANGPTL8,VCAM-1 and their combination on the occurrence of CVS in patients undergoing intracranial aneurysm embolization.Results The differences between CVS and non-CVS groups were statistically significant in hypertension,Hunt-Hess grade,and Glasgow coma(GCS)scores(P＜0.05).The serum ANGPTL8 and VCAM-1 levels in the CVS group were significantly higher than those in the non-CVS group(P＜0.05).There was a positive correlation between serum ANGPTL8 and VCAM-1(r=0.468,P＜0.05).Multivariate analysis showed that high level of ANGPTL8(OR=3.652,95％CI:1.434-9.302),high level of VCAM-1(OR=2.619,95％CI:1.212-5.658),Hunt Hess grade Ⅲ-Ⅳ(OR=1.927,95％CI:1.104-3.362),GCS score of 3-8(OR=2.813,95％CI:1.257-6.295)were independent risk factors for CVS in patients undergoing intracranial aneurysm embolization.The AUC of serum ANGPTL8 level in predicting CVS in patients undergoing intracranial aneurysm embolization was 0.844,and the cut-off value was 189.233 U/L;the AUC of serum VCAM-1 level in predicting CVS in patients undergoing intracranial aneurysm embolization was 0.795,and the cutoff value was 17.984 mg/L.The accuracy,sensitivity and specificity of the combined prediction for CVS were 89.81％,93.94％and 85.57％,respectively,which were obviously higher than those of the single prediction.Conclusion The serum levels of ANGPTL8 and VCAM-1 in CVS group are obviously higher than those in non CVS group.The combination of the two has a high predictive value for CVS after intracranial aneurysm embolization.