1.Effect of dexamethasone on biological characteristics of bone marrow stromal stemc ells
Jie LIU ; Zhengyi SUN ; Lei CAO
Chinese Journal of Orthopaedics 2000;0(11):-
Object To investigate the effect of dexamethasone(Dex)of different concentration on the differentiation of the bone marrow stromal stem cells(MSC).Methods MSC were isolated from mice and cultured in vitro.After the third passage,the cells were treated with Dex of different concentration,the expression of typeⅠcollagen mRNA(COL1mRNA),alkaline phosphatase mRNA(ALP mRNA)and adipocyte lipid-binding protein mRNA(ap2mRNA)were detected by quantitative RT -PCR technique.The ratio of the electrophoretic results of the ap2mRNA,COL1mRNA and the ALP mRNA with the house keeping gene of glyceraldehydes-3-phosphate dehydrogenase(GADPH mRNA)respectively,were used to show the relative content of the mRNA.Results COL1mRNA,ALP mRNA and ap2mRNA were ex pressed despite of the difference of the concentration of the Dex.When Dex was1?10 -7 mol/L,the COL1mRNA was0.47?0.12,and ap2mRNA was0.21?0.16;and when Dex was1?10 -8 mol/L,the COL1mRNA was0.96?0.17,and ap2mRNA was0.06?0.03.There was significant statistic difference be-tween the two groups respectively.But the expression of the ALP mRNA were0.35?0.13and0.46?0.24re spectively,there was no significant difference among them.Conclusion Dex could regulate the differenti-ation of the MSCs into adipocyte or osteoblast by regulating special gene expression.In addition,the differ-en tia tion of MSC induced by Dex was bidirectional depending on the concentration of the Dex.When the concentra tion of the Dex was high,it could induce MSC differentiating into adipocytes.This might be the molecular mechanism of the steroid induced osteonecrosis.And the results proved when Dex was1?10 -8 mol/L,it was suitable for inducing MSCs differentiation into osteoblasts.
2.Comparative study between mesenchymal stem cells derived form orofacial bone and those from long bone
Lin YUAN ; Han WANG ; Jin SUN ; Zhengyi YANG ; Lu HAO ; Guangsi PAN ; Yina CAO ; Jun QIAN
Journal of Practical Stomatology 2015;(4):526-530
Objective:To compare the proliferation and osteogenic differentiation between human bone marrow mesenchymal stem cells from orofacial bone(OMMSCs)and those from long bone(BMMSCs).Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.BMMSCs were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method.The surface markers of the cells were detected by flowcytometry.Single-colony formation,CCK assay and cell circle analyses were conducted.Osteogenic differentiation ability was evaluated by ALP activity test and Alizarin red staining after osteogenic induction culture.Results:The cell surface markers STRO-1 and CD105 of both stem cells were positive,CD34,CD31 and CD45 were negative. OMMSCs generated significantly higher numbers of colonies than BMMSCs.In addition,OMMSCs had a higher proliferation rate and more cells in proliferative(S +G2 )stage than BMMSCs.After osteogenic induction for 3,5,7 and 10 d,OMMSCs showed higher levels of ALP activity.OMMSCs formed significantly more mineralized nodules than BMMSCs after 21-day ostogenic induction.Conclusion:The proliferation and osteogenic differentiation capacity of OMMSCs are higher than those of BMMSCs.
3.Exosomes serected by mesenchymal stem cells derived form orofacial bone regulate the function of macrophage
Lin YUAN ; Yina CAO ; Zhengyi YANG ; Jin SUN ; Guangsi PAN ; Jun QIAN ; Jingjing SONG ; Han WANG
Journal of Practical Stomatology 2017;33(3):344-348
Objective:To investigate the trait of exosomes serected by mesenchymal stem cells derived form orofacial bone(OMMSCs-Exo) and the communication between the exosomes and macrophages.Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.Their cell surface markers were characterized by flow cytometry.the rate of colony formation and the differentiation potential of OMMSCs were evaluated.Exosomes were prepaired from the culture supernatants of OMMSCs(P4-P6).Transmission electron microscopy(TEM) and western blot were used to identify the exosomes.The expression of miRNAs associated with immunity such as miR-223 and miR-let-7c were determined by Real-time RT-PCR.Human peripheral blood mononuclear cells(PBMCs) were isolated from health donor and co-cultured with OMMSCs-Exo.After co-cultured for 24 h,the communication between exosomes and macrophages was tested using a confocal microscope.Results:Human OMMSCs were proved to have the characteristics of mesenchymal stem cells.The diameter of OMMSCs-Exo ranged from 40 to 160 nm.The OMMSCs-Exo expressed CD63 and CD81 and contained miRNAs associated with immune regulation such as miR223 and miR-let-7c.OMMSCs-Exo could be uptaken by macrophages.After co-culture of OMMSCs-Exo with marcrophages for 72 h,miR223 expression in macrophages increased.Conclusion:OMMSCs-Exo has the potential of immune regulation.
4.Acetyltransferase MORF regulates osteogenic differentiation potential of periodontal ligament stem cells
Lin YUAN ; Jin SUN ; Feng CHENG ; Zhengyi YANG ; Yina CAO ; Guangsi PAN ; Jun QIAN ; Enliang HE ; Han WANG
Journal of Practical Stomatology 2016;32(6):778-782
Objective:To compare acetyltransferase MORF level in periodontal ligament stem cells( PDLSCs) derived from healthy individuals ( H-PDLSCs) with those derived from the individuals with periodontitis ( P-PDLSCs ) . And to determine the effect of MORF on the osteogenic differentiation potential of PDLSCs. Methods: Human H-PDLSCs and P-PDLSCs were cultured and cloned with limited dilution method. H-PDLSCs were stimulated by LPS, TNF-α, IL-β and the mix of the 3 inflammatory factors to imitate inflammatory environment ( IP-PDLSCs ) . Quantitative RT-PCR and Western Blot were applied to examine different expression of MORF in H-PDLSCs and P-PDLSCs. Western Blot was applied to detect expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red staining were applied to determine osteogenic differentiation potential of H-PDLSCs with MORF knock-down. Results:Quantitative RT-PCR and Western Blot showed lower expression of MORF in P-PDLSCs compared with H-PDLSCs( P<0. 05). Western Blot revealed lower expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red stai-ning indicated osteogenic differentiation potential was inhibited in H-PDLSCs with MORF knockdown(P<0. 05). Conclusion: Peri-odontitis can suppress the expression of MORF in PDLSCs and inhibite the osteogenic differentiation potential of PDLSCs.
5.A multicenter prospective randomized open comparative study on the treatment of ovulatory menorrhagia with tranexamic acid and norethisterone in China
Yiwen ZHANG ; Fangfang HE ; Zhengyi SUN ; Shangwei LI ; Shiliang BI ; Xiuling HUANG ; Zansun CAO ; Shulan LU ; Junli Lü ; Zhenyu ZHANG ; Yimin ZHU ; Hefeng HUANG ; Maohua MIAO
Chinese Journal of Obstetrics and Gynecology 2008;43(4):247-250
Objective To compare the efficacy and safety of tranexamic acid(TA)and norethisterone(NET)for the treatment of patients with ovulatory menorrhagia in China. Methods Onehundred and thirty one patients with proven ovulatory menorrhagia from gynecologic clinics of 5 teaching hospitals located in 4 different cities in China were enrolled during Jul 2004 to Dec 2006.Ameng them 128 completed the study.Patients were randomly divided into two therapeutic regimen groups:TA 1g thrice daily during menstrual cycle days(D)1-5,69 cases;or NET 5 mg twice daily on D19-26.59 cases.The drugs were administered for 2 consecutive cycles,then withdrawn and patients were followed-up for 1 more cycle.Data on menstrual blood loss [ estimated by pictorial blood assessment chart(PBAC)],length of menstrual periods,quality of life(QOL)evaluated by a 6 item health-related questionnaire were collectedbefore,during each cycle and were compared.Results Both treatments led to significant decreases of mean PBAC scores and shorter duration of menstrual periods,and improved the QOL ranking during the twotreatment cycles.The mean percentages of PBAC decrements in the TA first and second cycles were significantly greater than those in the NET corresponding cycles(35%VS 17%,P=0.004;4J4%VS 34%,P=0.04 respectively).The success rate of TA second cycle was higher than that of the NET second cycle (41%VS 24%,P=0.04).Improvement of QOL ranking in the TA first cycle was also significantly better than those in the NET first cycle ( P=0.03).The percentage of patients with at least 1 adverse event in TA group(19%)was significantly lower than that in NET group(35%,P=0.04).Patients'willingness tocontinue the treatment in the TA second and follow-up cycles(94%,79%respectively)were significantly higher than those in the corresponding cycles of NET groups(79%,59%respectively;P=0.01,P=0.02).Conclusion The regimen of TA 3 g daily during menstrual days 1-5 is a more effective and tolerable treatment than luteal phase norethisterone for patients with ovulatory menorrhagia.
6.Role of MDA in the pathogenesis of GERD esophagus mucosal inflammatory injury
Mijiti ABULAJIANG ; Yisireyili MAIMAITI ; Aili AIKEBAIER ; Wulamu WUBULIKASIMU ; Zhengyi CAO ; Yiliang LI ; Alimujiang AZIGULI ; Yuan JIANG ; Abudureyimu KELIMU
Journal of Chinese Physician 2019;21(7):997-1001,1006
Objective To investigate the expression of malondialdehyde ( MDA) in esophageal mu-cosa of different types of gastroesophageal reflux disease ( GERD) patients and its role in the esophageal in-flammation. Methods According to the inclusion and exclusion criteria, 42 patients hospitalized in the the Xinjiang Uygur Autonomous Region People's Hospital from December 2017 to October 2018 were selected as the research group. 8 healthy subjects completed physical examination were set up as healthy control group. GERD completed GERDQ score, 24 h pH monitoring, and taken 3 cm on the dentate line of the esophagus as a specimen. The study group was divided into non-erosive reflux disease (NERD) group (17 cases) and Ero-sive reflux disease [erosive esophagitis (RE)] group (25 cases). Then hematoxylin-eosin (HE) staining, immunohistochemistry, real-time polymerase chain reaction ( qPCR ) , enzyme-linked immunosorbent assay (ELISA) methods were used to detect inflammation, oxidative stress (MDA), antioxidant enzyme [manga-nese superoxide dismutase (Mn SOD), glutathione (GSH), catalase (CAT)], and proinflammatory cyto-kines [monocyte chemotactic protein-1 (MCP-1), interlukin-8 (IL-8), tumor necrosis factorα(TNF-α)]. Results There was no significant difference in body mass index ( BMI ) between the three groups ( P >0. 05). 24 h pH monitoring of esophagus showed that the indexes of weak acid reflux (4