Objective To investigate the effect of osteoblast-derived exosome(Exo)mediating microRNA(miR)-494 on bone metabolism and bone remodeling balance in osteoporosis(OP)rats and its mechanism.Methods Exosomes was isolated and identified from MC3T3-E1 osteoblast cell line and transferred to Exo by electrical transfer.Forty SD rats were randomly divided into control,model,miR-494 mimic(miR-494),and miR-494 inhibitor group,10 rats in each group.The ovaries were removed to construct the OP model except the control group.After modeling,the miR-494 group and miR-494 inhibitor group received tail vein injections of exosomes containing the corresponding miRNA,at a dose of 3×109 particles.Four weeks later,bone parameters were detected in each group of rats by Micro-CT,serum bone markers were measured by ELISA,pathological changes in bone tissue were observed by HE staining,osteoclast numbers were detected by tartrate-resistant acid phosphatase(TRACP)staining,and the expression levels of bone remodeling-related proteins and toll-like receptor 4(TLR4)pathway-related proteins were determined by Western blotting.Results Typical cup-shaped or round exosomes were successfully isolated with a diameter of about 100 nm from MC3T3-E1 cells,which contained CD63,CD9,tumor susceptibility gene 101(TSG101),heat shock protein 70(HSP70)proteins and can be taken up by MC3T3-E1 cells.Compared with the model group,the bone parameters of the rats in the miR-494 mimic group decreased,serum bone markers bone Gla protein(BGP),TRACP,C-terminal telopeptide of type Ⅰ collagen(CTX-Ⅰ)increased,osteoprotegerin(OPG),procollagen type Ⅰ N-terminal propeptide(PⅠNP)decreased,bone trabeculae structure was disordered,osteoclasts increased,bone morphogenetic protein 2(BMP-2),Runt related transcription factor 2(RUNX2)in bone tissue downregulated,receptor activator of nuclear factor kappa-B ligand(RANKL)upregulated,TLR4,nuclear factor kappa-B p65(NF-κB p65)and myeloid differentiation primary response 88(MyD88)upregulated(all P＜0.05).In contrast,the situation of the miR-494 inhibitor group was opposite,bone parameters and OPG,PⅠNP increased,BGP,TRACP,CTX-Ⅰdecreased,bone structure returned to normal,osteoclasts decreased,BMP-2,RUNX2 in bone tissue upregulated,RANKL downregulated,TLR4,NF-κB p65 and MyD88 downregulated(all P＜0.05).Conclusion The transfer of miRNA-494 by Exo aggravates abnormal bone metabolism in OP rats and inhibits bone remodeling balance,suggesting that the mechanism of action may be related to the regulation of TLR4 pathway.

Objective: To investigate the association of the levels of high sensitivity C-reactive protein (hs-CRP) with frailty and its components among the elderly over 65 years old in 9 longevity areas of China. Methods: Cross-sectional data from the Health Ageing and Biomarkers Cohort Study (HABCS, 2017-2018) were used and the elderly over 65 years old were included in this study. Through questionnaire interview and physical examination, the information including demographic characteristics, behavior, diet, daily activity, cognitive function, and health status was collected. The association between hs-CRP and frailty and its components in the participants was analyzed by multivariate logistic regression model and restrictive cubic spline. Results: A total of 2 453 participants were finally included, the age was (84.8±19.8) years old. The median hs-CRP level was 1.13 mg/L and the prevalence of frailty was 24.4%. Compared with the low-level group (hs-CRP<1.0 mg/L), the OR (95%CI) value of the high-level group (hs-CRP>3.0 mg/L) was 1.79 (1.35-2.36) mg/L. As for the components, the hs-CRP level was also positively associated with ADL disability, IADL disability, functional limitation and multimorbidity. After adjusting for confounding factors, compared with the low-level group, the OR (95%CI) values of the high-level group for the four components were 1.68 (1.25-2.27), 1.88 (1.42-2.50), 1.68 (1.31-2.14) and 1.39 (1.12-1.72), respectively. Conclusion: There is a positive association between the levels of hs-CRP and the risk of frailty among the elderly over 65 years old in 9 longevity areas of China. The higher hs-CRP level may increase the risk of frailty by elevating the risk of four physical functional disabilities, namely ADL disability, IADL disability, functional limitation and multimorbidity.
Humans ; Aged ; Aged, 80 and over ; C-Reactive Protein/analysis* ; Frailty/epidemiology* ; Cohort Studies ; Cross-Sectional Studies ; China/epidemiology*

OBJECTIVE: To observe the clinical significance of translocator proteins (TSPO) gene in the treatment of FLT3-ITD/DNMT3A R882 double-mutated acute myeloid leukemia (AML). METHODS: Seventy-six patients with AML hospitalized in the Department of Hematology of the Affiliated People's Hospital of Ningbo University from June 2018 to June 2020 were selected, including 34 patients with FLT3-ITD mutation, 27 patients with DNMT3A R882 mutation, 15 patients with FLT3-ITD/DNMT3A R882 double mutation, as well as 19 patients with immune thrombocytopenia (ITP) hospitalized during the same period as control group. RNA was routinely extracted from 3 ml bone marrow retained during bone puncture, and TSPO gene expression was detected by transcriptome sequencing (using 2-deltadeltaCt calculation). RESULTS: The expression of TSPO gene in FLT3-ITD group and DNMT3A R882 group at first diagnosis was 2.02±1.04 and 1.85±0.76, respectively, which were both higher than 1.00±0.06 in control group, but the differences were not statistically significant (P=0.671, P=0.821). The expression of TSPO gene in the FLT3-ITD/DNMT3A R882 group was 3.98±1.07, wich was significantly higher than that in the FLT3-ITD group and DNMT3A R882 group, the differences were statistically significant (P=0.032, P=0.021). The expression of TSPO gene in patients who achieved complete response after chemotherapy in the FLT3-ITD/DNMT3A R882 group was 1.19±0.87, which was significantly lower than that at first diagnosis, and the difference was statistically significant (P=0.011). CONCLUSION TSPO gene may be used as an indicator of efficacy in FLT3-ITD /DNMT3A R882 double-mutated AML.
Humans ; DNA (Cytosine-5-)-Methyltransferases/genetics* ; DNA Methyltransferase 3A ; Mutation ; Leukemia, Myeloid, Acute/drug therapy* ; Nucleophosmin ; Prognosis ; fms-Like Tyrosine Kinase 3/genetics* ; Receptors, GABA/therapeutic use*

PTH-related peptide (PTHrP) improves the bone marrow micro-environment to activate the bone-remodelling, but the coordinated regulation of PTHrP and transforming growth factor-β (TGFβ) signalling in TMJ-OA remains incompletely understood. We used disordered occlusion to establish model animals that recapitulate the ordinary clinical aetiology of TMJ-OA. Immunohistochemical and histological analyses revealed condylar fibrocartilage degeneration in model animals following disordered occlusion. TMJ-OA model animals administered intermittent PTHrP (iPTH) exhibited significantly decreased condylar cartilage degeneration. Micro-CT, histomorphometry, and Western Blot analyses disclosed that iPTH promoted subchondral bone formation in the TMJ-OA model animals. In addition, iPTH increased the number of osterix (OSX)-positive cells and osteocalcin (OCN)-positive cells in the subchondral bone marrow cavity. However, the number of osteoclasts was also increased by iPTH, indicating that subchondral bone volume increase was mainly due to the iPTH-mediated increase in the bone-formation ability of condylar subchondral bone. In vitro, PTHrP treatment increased condylar subchondral bone marrow-derived mesenchymal stem cell (SMSC) osteoblastic differentiation potential and upregulated the gene and protein expression of key regulators of osteogenesis. Furthermore, we found that PTHrP-PTH1R signalling inhibits TGFβ signalling during osteoblastic differentiation. Collectively, these data suggested that iPTH improves OA lesions by enhancing osteoblastic differentiation in subchondral bone and suppressing aberrant active TGFβ signalling. These findings indicated that PTHrP, which targets the TGFβ signalling pathway, may be an effective biological reagent to prevent and treat TMJ-OA in the clinic.
Animals ; Osteoclasts ; Osteogenesis ; Parathyroid Hormone-Related Protein/pharmacology* ; Temporomandibular Joint ; Transforming Growth Factor beta/pharmacology*

To explore the medication rules of famous veteran traditional Chinese medicine practitioners in treating infertility based on medical cases of infertility collected from book series of Hundred Traditional Chinese Medicine Clinicians of Hundred Years in China and Prescription Proven by Traditional Chinese Medicine Masters. Researchers extracted the information of prescriptions from these cases according to the inclusion and exclusion criteria. Then, Excel 2010, SPSS Clementine(ver.12.0) and SPSS(ver. 22.0) were adopted respectively for frequency analysis, association rules analysis, cluster analysis and factor analysis. Cluster analysis was carried out by Ochiai algorithm of binary variable data, which was a systematic clustering method. And principal component analysis was used for factor analysis. Besides, KMO test and Bartlett spherical test were used for factor adaptation test. Finally, 151 medical cases and 396 prescriptions were included in total. A total of 60 kinds of frequently used herbs were identified according to the results of frequency analysis for medication, they were mainly used for activating blood and resolving stasis, tonifying and clearing heat respectively. The association rules analysis found out 25 drug pair association rules and 14 3-drug combination association rules. A total of 15 medicine groups were extracted by cluster analysis. KMO test and Bartlett spherical test indicated that the method was suitable for factor analysis, and 21 common factors were respectively extracted by factor analysis. Association rules indicated the characteristics of the therapeutic methods, like tonifying Qi and replenishing blood. The famous veteran traditional Chinese medicine practitioners utilized modified Siwu Decoction for tonifying blood and preferred Atractylodis Macrocephalae Rhizoma(Baizhu) for tonifying Qi. The results of both cluster analysis and factor analysis demonstrated the characteristics of the therapies for tonifying kidney, activating blood, tonifying spleen and dispelling dampness. In addition, factor analysis could reflect the therapies for nourishing Yin, tonifying kidney, warming the meridian, dissipating cold, nourishing blood and dispelling blood stasis. These results of analysis comprehensively showed out the medication characteristics of famous veteran traditional Chinese medicine practitioners of strictly following the pathogenesis, making good use of classical formulas and providing proper compatibility. In conclusion, data mining techniques(including frequency analysis, association rules analysis, cluster analysis and factor analysis) were used to comprehensively analyze the medication rules of famous veteran traditional Chinese medicine practitioners in treating infertility, which is helpful for guiding the clinical practice of treating infertility with traditional Chinese medicine.
China ; Humans ; Infertility ; drug therapy ; Medicine, Chinese Traditional

Objective To investigate the influencing factors of the formation of intracranial collateral circulation in patients with anterior circulation chronic occlusion. Methods From January 2015 to December 2017,181 consecutive patients with unilateral internal carotid artery or middle cerebral artery chronic occlusion diagnosed by DSA and admitted to the Department of Neurology,Chinese People′s Liberation Army General Hospital were enrolled retrospectively. According to the American society for interventional and therapeutic neuroradiology/society of interventional radiology ( ASITN/SIR ) collateral circulation grading system, 68 patients were divided into poor collateral circulation group (grade 0-2) and 113 were divided into good collateral circulation group (grade 3-4). After admission,the patients completed the relevant examinations, including blood routine,blood uric acid,blood lipids,and DSA examination. The age,gender,basic diseases (hypertension,diabetes,hyperlipidemia, coronary heart disease, history of previous ischemic stroke) and history of smoking/alcohol of the patients were recorded. The formation of collateral circulation was used as the dependent variable. The factor of P<0. 05 in the univariate analysis was included in the multivariate logistic regression analysis. Results (1) The patients of the poor collateral circulation group were older than those of the good collateral circulation group (61 ± 9 years vs. 56 ± 12 years),and the proportion of hyperlipidemia was higher than that of the good collateral circulation group (26. 5%[18/68] vs. 13. 3%[15/113]). The differences were statistically significant between the groups (all P<0. 05). There were no significant differences in the proportions of gender,hypertension,diabetes,coronary heart disease,ischemic stroke,tobacco and alcohol history between the two groups ( all P>0. 05) . (2) Compared with the good collateral circulation group,the level of high density lipoprotein ( HDL) in the poor collateral circulation group was lower. The differences were statistically significant between the groups (1. 0 ± 0. 2 mmol/L vs. 1. 1 ±0. 3 mmol/L,P<0. 05). There were no significant differences in blood uric acid,triglyceride,cholesterol, low-density lipoprotein,and platelet count levels between the two groups (all P>0. 05). (3) After variable selection,age (from young to old) and HDL (from low to high) as independent variables and the formation of collateral circulation as dependent variables, they were included in multivariate logistic regression analysis. The results showed that increasing age increases the risk of poor collateral circulation (OR,1. 053, 95%CI 1.021-1.085,P <0.05);the elevated HDL level was a protective factor of the formation of collateral circulation (OR,0. 265,95% CI 0. 085-0. 825,P<0. 05). Conclusion With the increase of age,the risk of intracranial poor collateral circulation increases,and the increased HDL level is beneficial to the formation of collateral circulation.

Objective: To investigate the impact of FLT3-ITD and DNMT3A R882 double mutations to the prognosis of acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: FLT3-ITD, DNMT3A, C-kit, CEBPA, FLT3-TKD and NPM1 mutations were detected in 206 newly diagnosed AML patients by Sanger sequencing (M(3) and those received FLT3 inhibitor were excluded). Clinical data of AML patients were retrospectively analyzed to compare the prognosis of each gene mutation group. Results: ①Of 206 patients, 104 were male and 102 female with a median age of 38 (3-63) years, including 6 cases of M(0), 24 cases of M(1), 56 cases of M(2), 39 cases of M(4), 63 cases of M(5), 6 cases of M(6) and 12 unclassified cases. ②All 206 patients were divided into four groups according to the mutation gene at the time of diagnosis: FLT3-ITD(+) DNMT3A R882(+) group (group A), FLT3-ITD(+) DNMT3A R882(-) group (group B), FLT3-ITD(-) DNMT3A R882(+) group (group C) and FLT3-ITD(-) DNMT3A R882(-) groups (group D). Gender, leukocyte count at diagnosis, chromosome karyotype, the median age, FAB classification, disease status prior to transplantation, type of donor, conditioning regimen and GVHD were not significantly different between four groups (P>0.05). ③The 2-year cumulative recurrence rate (CIR) of group A was significantly higher than that of other groups [group A (72.2±2.6)%, group B (38.6±0.6)%, group C (36.8±1.6)%, group D (27.8±0.1)%, respectively, P<0.05], while the 2-year overall survival (OS) rate and 2-year leukocyte-free survival (LFS) rate were lower than those of other groups [group A (30.9±13.3)%, (11.3±10.2)%; group B (67.5±7.8)%, (47.9±8.4)%; group C (61.4±12.4)%, (56.8±12.5)%; group D (80.1±3.7)%, (79.7±3.6)%, respectively, P<0.05]. Conclusion: AML patients with FLT3-ITD and DNMT3A R882 double mutations had a very high CIR and low OS, LFS after transplantation.
Adolescent ; Adult ; Child ; Child, Preschool ; DNA (Cytosine-5-)-Methyltransferases/genetics* ; DNA Methyltransferase 3A ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute ; Male ; Middle Aged ; Mutation ; Nucleophosmin ; Prognosis ; Retrospective Studies ; Young Adult ; fms-Like Tyrosine Kinase 3/genetics*

Objective To investigate the risk assessment,guiding role,and clinical value of Vaso CT image features for recanalization of vertebrobasilar junction occlusion. Methods From January 2016 to May 2017,14 patients with vertebrobasilar junction occlusion admitted to the Department of Neurology, Chinese PLA General Hospital were analyzed retrospectively. Preoperative cerebral angiography confirmed vertebrobasilar junction occlusion. Vaso CT was used to measure the length of the occluded vessels and vascular direction at both ends. According to these results, the operative risks were evaluated and the recanalization of vertebrobasilar junction occlusion were guided. Results The length of vertebrobasilar junction occlusion was 2. 56-19. 09 mm (mean 4. 5 ± 2. 1 mm) in 14 patients,and 13 of them were treated with the recanalization of vertebrobasilar artery occlusion,the blood vessels of 12 cases were successfully recanalized and stent placement was performed after the recanalization,among them,8 Solitaire stents and 4 Wingspan stents were implanted;One patient did not perform recanalization because of longer length of occlusion (19. 09 mm). All patients operated did not have any perioperative complications, and the neurological symptoms were significantly improved after procedure. Conclusion Vaso CT can accurately determine the surgical risk of the recanalization of vertebrobasilar junction occlusion,and can guide the surgical pathways,reduce the incidence of perioperative complications,and improve the success rate of the operation.

Adipocytes ; pathology ; Angiomyoma ; pathology ; Humans ; Nasal Cavity ; Nose Neoplasms ; pathology

Several studies have shown that the tumor endothelial cells are different from the normal tissue endothelial cells. These tumor endothelial cells may contribute to tumor neo-vasculogenesis. This study was purposed to analyze the biologic features and determine the expression level of CD133 and BCR/ABL fusion gene in circulating endothelial cells (CEC) isolated from peripheral blood of CML patients, as well as to investigate the role of CEC in disease progression. Mononuclear cells were isolated from peripheral blood by density gradient centrifugation; CEC were sorted by MACS and harvested in the endothelial growth medium. The morphologic features of CEC were observed by microscopy, the cell growth rate was calculated by cell counting, and the cells were identified by immunofluorescence staining for the expression of CD31,CD34,VWF and CD133. The expression of BCR/ABL fusion gene was examined by FISH in 12 CML patients. The results indicated that the isolated CEC displayed the typical cobble-stone morphology. These cells could be identified by the positive immunofluorescence staining for CD31, CD34 and VWF, and showed more increased proliferative potential as compared to that of healthy donors. It was found that the positive rate of CD133 was 31.29% in CML patients, which was significantly different from that of healthy donors (P < 0.05). In 12 CML patients, CEC carried the same chromosome aberration as the leukemia cells (10.77%). Higher expression level of CD133 and BCR/ABL fusion gene positively correlated with progression of disease. It is concluded that the CEC may participate in invasion and angiogenesis in patients with CML and possibly correlate to the spreading and progression of the disease.
AC133 Antigen ; Adult ; Antigens, CD ; metabolism ; Cell Proliferation ; Endothelial Cells ; metabolism ; Female ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Glycoproteins ; metabolism ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Male ; Middle Aged ; Neovascularization, Pathologic ; Peptides ; metabolism ; Young Adult