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Objective To explore the role of contact system activation in the mechanism of systemic lupus erythematosus (SLE) patients with thrombotic events.Methods A simple sample drawing study was conducted.Sixty-nine patients with SLE admitted to Department of Rheumatism in Tianjin Hospital from June 2014 to February 2016 were enrolled.The patients were divided into simple SLE group (n =38) and SLE + vascular diseases (VD) group (n =31) according to whether the patients complicated with VD or not.The VD patients were subdivided into three subgroups including SLE complicated with myocardial infarction (SLE + MI,n =10),SLE complicated with deep vein thrombosis (SLE + DVT,n =13),and SLE complicated with arterial thrombosis (SLE + AT,n =8).Sixty-eight healthy age and gender-matched volunteers without history of VD were served as controls.Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of FⅫa-C1 inhibitor (FⅫa-C1INH) and FⅫa-antithrombin (FⅫa-AT) in plasma.Flow cytometry was used to analyze the contents of platelets associated factors.The correlation between platelet associated factor and FⅫA-C1INH and FⅫa-AT was analyzed by Spearman correlation analysis.Receiver operating characteristic curve (ROC) was plotted to analyze the predictive value of FⅫA-C1INH and FⅫa-AT for SLE thrombotic events.Results Compared with health control group,the expression of FⅫa-C1INH in plasma in SLE group was significantly decreased [nmol/L:0.00 (0.00,0.07) vs.0.08 (0.03,0.13),P ＜ 0.01],the expression of FⅫa-AT was significantly up-regulated [nmol/L:0.18 (0.07,0.38) vs.0.16 (0.12,0.26),P ＜ 0.05].Compared with the simple SLE group,the expression of FⅫa-C1INH in SLE + DVT and SLE + AT groups was significantly decreased [nmol/L:0.03 (0.02,0.07),0.02 (0.01,0.04) vs.0.07 (0.02,0.11),both P ＜ 0.05],and the expression of FⅫa-AT in plasma in SLE + AT group was significantly increased [nmol/L:0.34 (0.21,0.53) vs.0.17 (0.06,0.30),P ＜ 0.01].It was shown by correlation analysis that FⅫa-C1INH was negatively related with FⅫa-AT in patients with SLE (r =-0.24,P =0.041 6).Activated platelet associated factors such as the production of interferon mediated by transmembrane protein 1 (IFTMI1) and interferon induced by double stranded RNA dependent activation agent (PRKRA) were positively related with up-regulation of FⅫa-AT and down-regulation of FⅫa-C1INH (IFITM1 and FⅫa-AT:r =0.39,P =0.001 2;IFITM1 and FⅫa-C1INH:r =-0.30,P =0.0146;PRKRA and FⅫa-AT:r =0.29,P =0.017 6;PRKRA and FⅫa-C1INH:r =-0.36,P =0.0029).The thrombospondin-1 (TSP-1) and platelet P-selectin were positively related with up-regulation of FⅫa-AT (r1 =0.72,P1 ＜ 0.0001;r2 =0.34,P2 =0.003 8).It was shown by ROC curve analysis that the area under the ROC curve (AUC) for FⅫa-C1INH on evaluating the risk of SLE thrombotic events was 0.998,the sensitivity was 100％,and specificity was 97.4％ when cut-off ＜ 0.01 nmol/L;AUC for FⅫa-AT for evaluating the risk of SLE thrombotic events was 0.954,the sensitivity was 95.0％,and specificity was 84.2％ when cut-off ＞ 0.40 nmol/L;predicted probability of two markers for predicting diagnosis was 0.5,the sensitivity and specificity were both 100％.Conclusions Contact system is activated in patients with SLE.FⅫA-C1INH and FⅫa-AT levels are closely related with platelet associated factors IFITM1 and PRKRA.FⅫA-C1INH and FⅫa-AT can be served as a promising potential biomarker for evaluation of the risk of thrombotic events in SLE.
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ObjectiveTo study the method of cell isolation,primary culture and identification of subchondral bone cell of osteoarthritis(OA) rabbits.Methods The rabbit instable knee joint models were made by modified Hulth modeling method.The osteoblasts were harvested from the subchondral bone of rabbits by collagenase and tissue explants attachment.The morphology observation and biological identification were performed by inverted microscope and immunocytochemistry staining,respectively.The proliferative activity of cells were detected by MTT and the expression of Ⅰ-collagen at gene level was detected.ResultsThe cells started to appeared on the 11th day after the attachment.The cells form were fusiformis and triangle,the nucleolus were clear.The cultured cells had typical osteoblast morphological characteristics.The cells obtained from subchondral bone of rabbits were identified to be osteoblast by immunocytochemistry staining.The proliferative activity of cells were equably proliferation which detected by MTT.ConclusionThe modified method provides better way to obtain ideal subchondral osteoblast and the co-culture method is suitable for the study of OA microenvironment,which can simulate interactions of the subchondral osteoblast,synovial cells and chondrocyte.
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BACKGROUND: The Chinese medicine, Sheng-Ji Ointment, is utilized to cure bone defect due to infectious open fractures in the clinical field. So it was imagined that it was a new inductive factor of osteogenesis and its ingredients could acceleate the proliferation of bone marrow-derived mesenchymal stem cells (BMSCs) and induce them into osteoblasts. In our primary experiment, the Sheng-Ji liquor, which was extracted from the same Chinese herbs with Sheng-Ji Ointment, had been identified that it affected the proliferation of BMSCs with its different concentration.OBJECTTVE: To investigate the effect of Sheng-Ji liquor on the osteogenesis of in vitro cultured BMSCs from rabbit tibia.DESTGN: An observation in single kind of sample.SETTING: Orthopaedic Research Institute of Tianjin Hospital.MATERIALS: The experiment was carried out in the Tissue Engineering Laboratory of Tianjin Orthopadic Institute of Integrated Chinese and Western Medicine from October 2004 to December 2005. BMSCs were obtainedfrom male healthy Japanese long-eared rabbits of 3 months old, (3.5±0.5) kg. Sheng-Ji liquor (5×102 g/L) was extracted from the Chinese herbs with Sheng-Ji Ointment (Tianjin Factory of Chinese Crude-drug and Cut Crude-drug), including the crude drugs of angelica, rehmannia dride rhizome, carapax testudinis, corium elephatis and crinis carbonisatus.METHODS: The Sheng-Ji liquor was put to BMSCs in different concentrations of 25, 8.3, 5 g/L. Basic medium without Sheng-Ji liquor was taken as blank control, while the standard-controlled medium contained the ingredients of the following blank-controlled medium, dexamethasone 10-8 mol/L, vitamin C 0.05 g/L and β-sodium glycerophosphate 10 mmol/L. The BMSCs passaged in different conditions were seeded in plate and cultured for 2-3 weeks, then the mineralized nodes were observed under inverted fluorescence microscope, and the cell expressions were observed with acheomycin labeling and collegan Ⅰ immunohistochemical staining. The culture medium was collected to determine the contents of extracellular and intracellular alkaline phosphatase (ALP), osteocalcin (OCN) and lactate dehydrogenase (LDH), then the ratios of ALP/LDH and OCN/LDH were calculated.MAIN OUTCOME MEASURES: After 2 and 3-week culture, the mineralized nodes, results of acheomycin labeling and collegan Ⅰ immunohistochemical staining, and the ratios of ALP/LDH and OCN/LDH were observed.were prolonged, proliferated assembly and overlapped. The inter-cell limits disappeared gradually and sporadic cellular Results of acheomycin labeling and collegan Ⅰ immunohistochemical staining: There were positive cells and mineralized nodes in all of groups with Sheng-Ji liquor and standard-controlled group. BMSCs of the blank-controlled group were negative. The positive cells and mineralized nodes were green with yellowish color under irrigation of fluorescence; The results of immunocytochemistry for collagen Ⅰ were positive in all of groups with Sheng-Ji liquor and standard-controlled group. The result of blank-controlled group was negative. The positive induced BMSCs had the cytoplasm with granula of highest as compared with the other groups (P ＜ 0.05). The ratios of ALP/LDH and OCN/LDH in the other groups were close (P ＞ 0.05).
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Bone tissue engineering is one of the most clinical applicable research areas in tissue engineering. Seed cell is the first step and essential element of construction and application of tissue-engineered bone. In recent years, many inspiring achievements have been gained in the field of seed cell study. In this review, current status and the prospects of study of seed cell source in bone tissue engineering are reviewed.
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The research of bone tissue engineering provides new thought and method to repair mass bone defect. Neovascularization plays a significant role in bone repair. This article reviews the advancements of the growth factors, seed cells and scaffolds in vascularization of tissue engineering bone, then raise the problems to solve and the prospect of future research.
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Cell apoptosis is an important biological process. It plays an essential role on cell growth and response for outside stimulations. This review focused on the recent advancement of apoptotic signal transduction, enzymological mechanisms, the function of mitochondria in cell apoptosis as well as the regulation of genes.
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[Objective]To evaluate the results of therapy with knee joint synovectomy or articular irrigation in rheumatoid arthritis.[Method]Thirty-six knees in 32 rheumatoid arthritis patients-according to ARA standard were performed with knee joint synoveetomy and articular irrigation.Twenty-two knees in 20 patients(most in grade Ⅰ)were treated with knee joint cavity irrigation operation combined with intra-articular injection of hyaluronic acid,and 14 knees in 12 cases(most in grade Ⅱ)with knee joint synovectomy,All 32 patients received routine anti-rheumatoid drug therapy preoperatively and postoperatively,and were followed-up for 6 months.All the cases were evaluated by the Lysholm scale.[Result]The early symptoms of all knee joints were improved.The excellent and good result rate of articular irrigation-group was 86.4%,and in synovectomy-group was 85.7%.[Conclusion]Satisfactory results can be obtained through articular irrigation combined with intra-articular rejection of hyaluronic acid and concomitant anti-rheumatoid drug therapy in early-stage of rheonmtoid arthritis,but for mid-stage patients,especially with hyperplasia synovium and deslxoyed cartilage,joint synoveetom should he performed early in order to obtain favoring rehabilitation.
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Objective To study ADFR with statin programmed therapy of osteoporosis in ovariectomized rats.Methods Fifty female rats were randomly allocated into 2 groups:sham operation (S, n =10) and OVX ( n =40) group.After operation for one month,OVX were randomly allocated into 4 groups (each n =10):OVX,statins (T),bisphosphonates (B) and statins+bisphosphonates+calcium+vitamin D (ADFR).After feeding statins or bisphosphonates or ADFR for 100 days,all rats were sacrificed.The effects of T or B or ADFR on bone histomorphology or osteocalcin in sera or deoxypyridoxine in urea were studied.Results The data showed that osteocalcin and deoxypyridine in OVX group were significantly improved compared with S group ( P 0 05) ,in B group was decreased,and in ADFR group was increased compared with OVX group.The histomorphometric date showed that TOS,MOSW,STS/DTS,TBOS,TBSC and iMAR in OVX were significantly increased,and TBV,MLT and ? in OVX were decreased,compared with S group.TBV in B,T and ADFR groups was larger than that in OVX group.TOS,MOSW,TBOS and TBCS in B group were smaller than those in OVX group,? in B group was longer than that in OVX group,TBCS and ? in T group were increased compared with OVX group.Conclusion Statins promote bone turnover,increase osteoblast activity and osteoid production,and reduce the bone construction.Bisphosphonates inhibit bone absorption,while ADFR acelerate bone formation and reduce bone loss,suggesting that polytherapy is preferable to monotherapy.
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Objective To evaluate the clinical efficacy of domestic bisphosphonates (etidronate and alendronate) in the treatment of postmenopausal osteoporosis. Methods Eighty three patients with postmenopausal osteoporosis were divided randomly into three groups: group Ⅰ(37 cases),group Ⅱ(15 cases) and group Ⅲ(31 cases). Etidronate(400 mg/d) and placebo were given with intermittent cyclical therapy in group Ⅰ and group Ⅲ respectively, and alendronate(10 mg/d) was given continuously in groupⅡ for 9 months. Eeach group was given calcium agents(equal to elementary calcium 500 mg/d). Before and after treatment, the bone mineral density(BMD) was measured by dual energy X ray absorptiometry. The biochemical markers such as blood calcium, phosphate, alkaline phosphatase and urine calcium/creatinin were examined. Results After treatment the BMD of groupⅠ and groupⅡ wee increased obviously(P0.05). The BMD of group Ⅱ was increased more than that of groupⅠ(P