4.miR-21/Sprouty1 function axis regulates the osteogenic differentiation of bone marrow mesenchymal stem cells after postmenopausal osteoporosis
Nan YANG ; Wei ZHOU ; Guang WANG ; Yin DING ; Yan JIN
Chinese Journal of Tissue Engineering Research 2017;21(21):3287-3292
BACKGROUND:Osteogenic differentiation is a complex process involving transcriptional and post-transcriptional regulation by multiple signaling pathways, and the specific mechanisms remain unclear. It is of great significance to study the role of critical miRNAs in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in the treatment of osteoporosis and bone defects. OBJECTIVE: To explore the regulatory ability of miR-21/Sprouty1 function axis in the osteogenic differentiation of BMSCs in patients with postmenopausal osteoporosis. METHODS:BMSCs were isolated from healthy people (H-hBMSCs) and patients with postmenopausal osteoporosis (PMOP-hBMSCs), and their osteogenic ability was compared. Expression of miR-21 and Spry1 at gene and protein levels was detected by real-time RT-PCR and western blot assay, respectively. miR-21 expression was upregulated via transfection in PMOP-hBMSCs, and the osteogenic ability and Spry1 expression of the cells were detected, while real-time RT-PCR and western blot were used to detect the expression of osteogenic marker genes, Runx2 and Osterix. RESULTS AND CONCLUSION:Compared with H-hBMSCs, PMOP-hBMSCs osteogenic ability was weakened significantly, miR-21 expression decreased, and Spry1 expression increased, indicating an inhibition to the miR-21-Spry1 function axis. Through the transfection of miR-21 and down-regulation of Spry1, the expression levels of Runx2 and Osterix were increased, and PMOP-hBMSCs osteogenic ability was partially restored.
5.Role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells under miR-21 regulation
Nan YANG ; Wei ZHOU ; Guang WANG ; Yin DING ; Yan JIN
Chinese Journal of Tissue Engineering Research 2014;(32):5085-5090
BACKGROUND:Previous studies have found that miR-21 expression is increased during osteogenic differentiation of bone marrow mesenchymal stem cells, but the action and molecular mechanism of miR-21 are stil unclear. OBJECTIVE:To verify the target gene of miR-21, Spry1, and to explore the role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells. METHODS:Luciferase report was used to verify Spry1 gene targeted by miR-21, and western blot assay was used to detect the expression of Spry1 in the osteogenesis of human bone marrow mesenchymal stem cells. Spry1 expression vector was established and transfected into human bone marrow mesenchymal stem cells. Osteogenesis ability of human bone marrow mesenchymal stem cells was analyzed after Spry1 high expression by alkaline phosphatase, alizarin red staining, RT-PCR and western blot. RESULTS AND CONCLUSION:Luciferase report suggested that Spry1 was a target gene of miR-21. The expression level of Spry1 was decreased in the osteogenesis of human bone marrow mesenchymal stem cells. Increasing expression of Spry1 could inhibit osteogenic differentiation of human bone marrow mesenchymal stem cells. These results indicate that Spry1 as a target gene of miR-21 negatively regulates osteogenic differentiation of human bone marrow mesenchymal stem cells, and plays an important role in bone formation process.
6.Therapeutic Effect of Fludarabine,Cytarabine and Granulocyte Colony-Stimulating Factor Regime on Relapsed and Refractory Acute Leukemia in Children
wei, LIN ; xuan, ZHOU ; bin, WANG ; guang-hua, ZHU
Journal of Applied Clinical Pediatrics 2006;0(15):-
Objective To primarily explore the efficacy and adverse effects of the combination of fiudarabine,cytarabine and granulocyte colony-stimulating factor(G-CSF)(FLAG regime)therapy for relapsed and refractory acute leukemia in children.Methods Ten children were treated with the FLAG regime for relapsed and refractory acute myeloid leukemia (AML)and acute lymphoblastic leukemia(ALL)from Feb.2007 to Mar.2010.There were 8 male and 2 female,with mean age 8 years(ranging from 4 to 12 years).AML was diagnosed in 8 children,AML-M2 in 5 cases,AML-M4 in 3 cases.ALL was diagnosed in 2 children,both were B-ALL.Six children had refractory disease,and 4 cases were in relapse.FLAG regime included:fludarabine 25 mg?m-2?d-1,days 1-5;cytarabine 2 g?m-2?d-1,days 1-5;G-CSF 150-300 ?g?d-1,from day 0 to neutrophils ≥0.5?109 L-1.Results Complete remission was obtained in 6 children(60%),partial remission was obtained in 1 child(10%),and 3 children were considered non-response(30%).The total effective rate was 70%.For 8 children with AML,6 children had achieved complete remission(75%),2 children had non-response(25%).While in children with ALL,1 child got partial remission,and the other one had non-response.Myelosuppression and infections due to neutropenia were the most frequent adverse effects,severe nonhematologic toxicity were not observed in these children.And there were no chemotherapy-related death.Conclusions The FLAG regime is effective in treatment of children with relapsed and refractory acute leukemia,especially for the children with the relapsed and refractory AML.The adverse effects from this regime were well tolerated.FLAG regime can give children with relapsed and refractory acute leukemia another chance.
8.The Establishment of an Anti-Trypanosoma Drug Screening System with Leucyl-tRNA Synthetase as an Inhibition Target
Guang-Wei GAO ; Ying YAO ; Da-Zhong DING ; Long YE ; Hu-Chen ZHOU ; Da-Wei LI ;
China Biotechnology 2006;0(12):-
Trypanosoma is a human parasite severely affecting poor tropical areas.However,current frontline drugs for Trypanosoma treatment have severe side-effects with decreased effectiveness.Based on the fact that aminoacyl-tRNA synthetase is a bonafide drug target for several microorganisms,including bacteria and fungi,it is plausible that it may also be effective target of Trypanosoma.The Trypanosoma brucei leucyl-tRNA synthetase(tbLeuRS)was cloned,expressed and purified to develop an in vitro enzymatic assay system.The assay conditions were further optimized for the effective screening of tbLeuRS inhibitors thus establishing an anti-Trypanosoma drug screening system targeting tbLeuRS.The results indicated that this system can be employed for the effective screening of anti-Trypanosoma drugs with satisfactory specificity.In addition,this system can also be used for compound optimization,as well as IC50 testing.Using this system a series of compounds are identified that are effective Trypanosoma inhibitors without toxicity to human cells.Therefore,targeting tbLeuRS may represent a new venue for the development of anti-Trypanosoma drugs.
9.Extraction Process Optimization of Fufang Jinsha Lidan Granule by Orthogonal Experiments
Ailing YI ; Fusheng ZOU ; Bo YI ; Guang DU ; Conghui ZHOU ; Yingying WEI ; Wei CHEN
China Pharmacist 2016;19(10):1963-1965
Objective:To optimize the extraction technology of Fufang Jinsha Lidan granule. Methods:According to the properties of traditional Chinese medicines in the formula, orthogonal experiments were used to optimize the extraction process of Fufang Jinsha Li-dan granule. The volume of water, boiling time and boiling frequency were used as the three influencing factors with three different lev-els in the orthogonal experiments. Moreover, the content of paeoniflorin and the yield of extract were chosen as the evaluation indices. The orthogonal experiments were carried out according to the L9 (34 ) orthogonal table. Results:The optimal extraction process of Fu-fang Jinsha Lidan granule was as follows:boiled twice, and one hour per time with 12-fold amount of water ( soaked for 30 min with 14-fold amount of water for the first extraction process) . Conclusion:The extraction process has such properties as high extraction rate, stability, simple operation, high yield of extract and controllable quality, which is worthy of wide application.
10.Application of salivary cortisol test for diagnosis of Cushing′s syndrome
Wei-Wei ZHOU ; Wei-Qing WANG ; Guang NING ; Ting-Wei SU ; Lei JIANG ; Na ZHU ; Yu-Fang BI
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
A total of 121 subjects comprising 40 normal subjects,58 patients with overweight or obesity and 23 patients with Cushing's syndrome were recruited in the study.The modified radioimmunoassay (RIA) for salivary cortisol test was established'and its normal range was determined.Then the diagnostic value of the salivary cortisol for the initial diagnosis of Cushing's syndrome was evaluated and single midnight salivary cortisol test demonstrated a sensitivity of 100.0% and specificity of 91.4 %.Salivary cortisol test can be recommended as a first-line diagnostic parameter for Cushing's syndrome.