1.The Effect of Antisense miR-30d on Invasion and Migration in Breast Cancer Cells
Ge CHEN ; Ping LUO ; Yu GONG ; Yao ZHOU ; Ruijun ZHAO ; Chunwei XIE
Tianjin Medical Journal 2014;(3):203-207
Objective To investigate the expression of miR-30d in breast cancer tissues and demonstrate the regula-tive effects of miR-30d ASO on the invasion and migration of breast cancer cells in vitro. Methods The expression levels of miR-30d in 108 breast cancer tissues and their adjacent tissues were detected by real-time quantitative PCR method. Af-ter transfection with miR-30d ASO, the biological effects of miR-30d on in breast cancer cells was measured by transwell as-say and wound healing assay. The expressions of matrix metalloproteinase (MMP)-2 and MMP-9 were analyzed by Western blot assay. Results The expression level of miR-30d was found to be over-expressed in breast cancer tissues(P<0.05). Compared with control group and nonsense interference group, the miR-30d expression was significantly decreased in breast cancer cells(transfection with miR-30d ASO). Results of transwell and wound healing assay showed that the invasion and mi-gration ability decreased significantly after transfection with miR-30d ASO, and expressions of MMP-2 and MMP-9 were down-regulated (P<0.05). Conclusion miR-30d was over-expressed in human breast cancer. The invasion and migration of breast cancer cells can be effectively inhibited by decreasing the expression of miR-30d. miR-30d may become a new tar-get for the regulation of invasion and migration in breast cancer.
2.Biological characteristics of synovial mesenchymal stem cells after isolation and purification
Peiliang FU ; Lei ZHANG ; Yuli WU ; Haishan WU ; Ruijun CONG ; Song CHEN ; Zheru DING ; Qi ZHOU
Chinese Journal of Tissue Engineering Research 2013;(41):7221-7227
BACKGROUND:A large amount of studies have confirmed that synovial mesenchymal stem cells have the similarity in cellmorphology, immune phenotype, colony forming ability and differentiation potential with bone marrow mesenchymal stem cells. But bone marrow mesenchymal stem cells are better than synovial mesenchymal stem cells in the ability to differentiate into cartilages. OBJECTIVE:To discuss the possibility of using synovial mesenchymal stem cells as seed cells for meniscal tissue engineering. METHODS:The synovial mesenchymal stem cells were isolated from rabbit synovial tissues with limiting dilution monoclonal culture method, and then the cells were purified. The morphology, ultrastructure, molecular phenotype, proliferation kinetics, karyotype and tumorigenicity of the in vitro cultured cells were analyzed. RESULTS AND CONCLUSION:The synovial mesenchymal stem cells isolated from the rabbit synovial cells had high proliferation capacity during in vitro monolayer culture. The synovial mesenchymal stem cells grew to peak at 6 days, and the doubling time was (30.2±2.4) hours. Flow cytometry results showed the synovial mesenchymal stem cells could express some molecular makers of mesenchymal stem cells, such as CD44 and CD90. DNA contents check, karyotype test and oncogenicity test confirmed isolated and purified synovial mesenchymal stem cells were the normal diploid cells without tumorigenicity, so the cells can be used as seed cells for meniscal tissue engineering.
3.Effects of Yiqi-Fumai injection and Shenshao capsule on vascular endothelial cell function in patients with angina pectoris of coronary heart disease and serum myeloperoxidase
Dongyan WANG ; Guizhen ZHAO ; Guozhong ZHANG ; Fengling HE ; Peng CHEN ; Wenjiang ZHOU ; Ruijun CAI
International Journal of Traditional Chinese Medicine 2015;(4):319-323
Objective To observe the therapeutic effect ofYiqi-Fumai injection andShenshao capsule in the treatment of angina pectoris and the influence on vascular endothelial cell function and serum MPO. Methods 90 cases of angina pectoris of coronary heart disease were randomly divided into a treatment group and a control group. 50 cases in the treatment group were treated withYiqi-Fumai injection and Shenshao capsule; 40 cases in the control group were treated with Plavix and metoprolol; clinical symptoms were observed before and after treatment, the indexes of endothelial cell function and electrocardiogram blood serum nitric oxide (NO), endothelin (ET), NOS and serum MPO, SOD, GSH changes were compared.Results The content of nitric oxide (NO), 6-keto-PGFLa and NOS of the treatment group were increased than the control group, while ET was reduced. In the treatment group, NO was (78.09 ± 88.71)μmol/L 6-keto-PGFL was 32.04 ± 25.82 pg/ml and NOS was 27.88 ± 9.08 U/MI before the treatment,; after the treatment, the NO was 127.11 ± 103.60μmol/L, 6-keto-PGFLa was 50.03 ± 60.87 pg/ml and NOS was 35.12 ± 22.18 U/MI. In the control group, NO was 96.98 ± 82.45μmol/L, 6-keto-PGFLa was 40.31 ± 51.92 pg/ml and NOS was 28.12 ± 5.11 U/MI before the treatment; After the treatment, NO was 95.62 ± 6.31μmol/L, 6-keto-PGFLa was 29.09 ± 42.01 pg/ml and NOS was 30.11 ± 7.51 U/MI. In the treatment group, ET was 58.81 ± 33.18 pg/ml before the treatment and 42.47 ± 40.5 pg/ml. after the treatment. In the control group, ET was 56.81 ± 32.76 pg/ml before the treatment and 60.67 ± 12.81 pg/ml after the treatment. In terms of resistance to oxidative stress and MPO, the SOD ,GSH and MPO improvement rate of the treatment group was significantly better than the control group (P<0.05). In the treatment group, SOD was 48.36 ± 17.38 U/MI, GSH was 126.68 ± 19.93 g/L and MPO was 57.33 ± 8.34 mmol/L before the treatment, after the treatment, SOD was 62.45 ± 15.76 U/MI, GSH was 135.98 ± 28.99 g/L and MPO was 17.11 ± 3.60 mmol/L. In the control group, SOD was 48.44 ± 19.37 U/MI, GSH was 29.44 ± 21.66 g/L and MPO was 56.35 ± 14.44 mmol/L before the treatment, after the treatment, SOD was 57.35 ± 14.44 U/MI, GSH was 28.56 ± 24.06 g/L and MPO was 48.62 ± 6.31 mmol/L.ConclusionYiqi-Fumai injection combined with Shenshao capsule has curative effect on angina pectoris of coronary heart disease, being worth of clinical use.
4.Ultrasound study of brachial plexus
Wen CAO ; Ruijun GUO ; Yajing ZHOU ; Miao ZHANG ; Zexing YU ; Xiaoning LIANG
Chinese Journal of Ultrasonography 2008;17(4):336-338
Objective To explore the anatomic character of brachial plexus nerve by ultrasonography,and provide a basis for ultrasound-guided brachial plexus block.Methods Twenty-four healthy volunteers were chosen for high-frequency sonography examination,the features of brachial plexus distributions were studied in the axilla location and the interscalene location.Results Brachial plexus of all the 24 cases were detected.At the axilla level,main braches of brachial plexus nerves were very close to the axillary artery.At the interscalene level,the brachial plexus was consistently found between the scalenus anterior and medius muscles in the interscalene groove.Conclusions Using high-frequency sonography,the main braches of brachial plexus can be observed clearly.
5.Histocompatibility between small intestinal submucosa and synovial mesenchymal stem cells
Peiliang FU ; Lei ZHANG ; Yuli WU ; Haishan WU ; Ruijun CONG ; Song CHEN ; Zheru DING ; Qi ZHOU
Chinese Journal of Tissue Engineering Research 2013;(47):8202-8208
BACKGROUND:Smal intestinal submucosa is characterized as antimicrobial activity, good biocompatibility, bio-mechanical properties, and rapid degradation in vivo, similar to the extracellular matrix of meniscal
fibrochondrocytes.
OBJECTIVE:To observe whether there exists a good histocompatibility between smal intestinal submucosa and synovial mesenchymal stem cells.
METHODS:Smal intestinal submucosa was treated with physical and chemical treatment. And hematoxylin-eosin staining and scanning electron microscopy observation were performed. Then, smal intestinal submucosa extracts were prepared for the fol owing experiments. (1) Pyrogenic test:smal intestinal submucosa extracts were injected into the ear vein of New Zealand white rabbits. (2) Skin sensitization test:smal intestinal submucosa extracts, paraformaldehyde solution and normal saline were respectively injected intradermal y into New Zealand white rabbits. (3) General toxicity test:smal intestinal submucosa extracts and normal saline were respectively injected into the ear vein of New Zealand white rabbits. Smal intestinal submucosa was co-cultured with osteogenic rabbit synovial mesenchymal stem cells, and smal intestinal submucosa cultured alone served as control. RESULTS AND CONCLUSION:There were some intestinal mucosal epithelial cells, fat cells and other cells adhered onto the surface of smal intestinal submucosa after physical treatment. While, the amount of residual cells decreased sharply after chemical treatment. But the main structure and the component had not been changed. The surface of smal intestinal submucosa was smooth and no cells remained, and there was a three-dimension network spatial structure. The porosity was 80%. Smal intestinal submucosa is a non-toxic, nonirritating, non-immunogenic biomaterial with very good biocompatibility, which has a good histocompatibility with rabbit synovial mesenchymal stem cells.
6.Effects of angiotensin II blockade on renal blood flow and renal oxygen consumption in chronic renal failure rats
Tingting ZHU ; Desheng FAN ; Jing YANG ; Shanshan ZHOU ; Chen WANG ; Ruijun YAN ; Liqun HE
Acta Laboratorium Animalis Scientia Sinica 2014;(5):1-6
Objective To observe the effects of angiotensin Ⅱ( Ang Ⅱ) blockade on renal function, renal blood flow and renal oxygen consumption in chronic renal failure ( CRF) rats induced by 5/6th kidney ablation /infarction (5/6A/I).Methods Sprague-Dawley rats were randomly divided into 3 groups:the normal group (group A, n =14), mod-el group (group B, n=14) and angiotensin II blockade (Cozaar with Monopril) treatment group (group C, n =14).The chronic renal failure ( CRF) rat models were induced by 5/6th kidney ablation/infarction.The tail artery systolic pressure (SBP), diastolic blood pressure (DBP) and tail vein serum creatinine (Scr), blood urea nitrogen (BUN), hemoglobin ( Hb) and creatinine clearance rate ( Ccr) were assessed before and after intervention.The course of treatment was sixty days.The renal blood flow ( RBF) , blood gas analysis of abdominal aortic and renal vein, left renal vein pressure ( RV-pO2 ) were detected and remnant renal oxygen consumption ( QO2/TNa ) was calculated, and the pathological changes of remnant kidney were observed after the 60 d intervention.Results (1) Compared with the group A, the levels of Scr, BUN and tail artery SBP, DBP were significantly increased ( P<0.01 for all) , and the levels of Ccr and Hb were signifi-cantly decreased ( P<0.01) in the groups B and C, demonstrating the successful modeling.(2) Compared with the group B, the levels of Scr, tail artery SBP, DBP and QO2/TNa were significantly decreased (P<0.01 for all), the levels of BUN were decreased (P<0.05), the levels of Hb, Ccr and RVpO2 were significantly increased (P<0.01 for all), the level of RBF was increased ( P<0.05) in the group C after intervention.(3) The histopathological examination of the remnant re-nal tissue showed that the pathological changes in the group C were apparently reduced, better than those of the Group B. Conclusions Angiotensin II blockade can increase RBF, reduce renal oxygen consumption, improve renal function, and reduce the renal pathological changes in CRF rats.The mechanism of renal protection may be related to the regulation of cellular energy metabolism and improvement of renal oxygen consumption.
7.The correlation between cognitive impairment and anti N-methyl-D aspartate receptor antibody in patients with systemic lupus erythematosus
Yan ZHOU ; Ruijun ZHANG ; Yang LI ; Xiaofeng ZENG ; Zhenhai WANG ; Hong ZHU
Chinese Journal of Rheumatology 2016;20(5):312-316
Objective To investigate the serum level of anti N-methyl-D-aspartate (NMDA) receptor antibody in patients with cognitive dysfunction of systemic lupus erythematosus (SLE) and its clinical significance.Methods The levels of anti NMDA receptor antibody in blood samples from 36 patients with cognitive dysfunction of SLE,30 cases with normal cognitive function of neuropsychiatric lupus erythematosus (NPSLE) groups,64 patients with no-NPSLE,100 patients with other connective tissue diseases and 30 healthy controls were measured by enzyme linked immunosorbent assay,and the reports of Mini-Mental State Examination and the laboratory indexes were recorded in those groups,then these index were measured again when patients were treated for 2 weeks.The comparison of multi-group was conducted by means of single factor analysis of variance or rank test,the Pearson or Spearman correlation analysis was used to analyze the correlation between variables.Results ① The level of anti NMDA receptor antibody in the patients with cognitive dysfunction of the NPSLE group (1.30±0.27) was higher than those with normal cognitive function of NPSLE group (0.47±0.08),No-NPSLE group (0.47±0.07),the other connective tissue disease (0.42±0.04) and the healthy controls (0.35±0.06) (P<0.05),the difference was statistically significant.The levels of anti NMDA receptor antibody before treatment were higher than those after the treatment,and the difference was statistically significant (t =2.296,P<0.05).② The serum levels of anti NMDA receptor antibody in patients were negatively correlated with the intelligent score (r=-0.330,P<0.01),and were positively correlated with the levels of IgG (r=0.684,P<0.05).Conclusion The anti NMDA receptor antibody can be used as a new autoantibody in the early diagnosis of cognitive dysfunction in patients with SLE,and can be used for monitoring the activity of disease in cognitive dysfunction of SLE.
8.Experimental study on the compound culture of small intestinal submucosa with synovial mesenchymal stem cells
Song CHEN ; Song PENG ; Peiliang FU ; Yuli WU ; Zheru DING ; Qi ZHOU ; Ruijun CONG ; Haishan WU ; Zhenyu XU
Chinese Journal of Orthopaedics 2014;(10):1059-1067
Objective To investigate the biosecurity and biocompatibility of small intestinal submucosa (SIS) as scaffold for tissue engineering and to explore the possibility to induce synovial mesenchymal stem cells (SMSCs) differentiate into cartilage with SIS as the scaffold and SMSCs as the seed cells. Methods The SMSCs were isolated and cultured from the synovial mem-brane of knee joints of rabbits by a conventional method. The SIS was harvested from pigs according to the physical method and Abraham's method. 4 rabbits are divided into the experimental group and control group. The biosecurity of SIS as scaf-folds were investigated in pyrogen test, skin sensitization test and systemic toxicity test. The SMSCs and SIS were co-cultured in vitro and induced to differentiate into cartilage to explore the biocompatibility of SMSCs and SIS, the proliferation and differ-entiation ability of SMSCs on the scaffold of SIS. The varietyies were identified by the microscope. Results The SIS isolated with the physical method and Abraham's method is a milky and translucent membrane with a smooth surface. Under the electron microscope, SIS presented a porous Stereoscopic three-dimensional network structure, which is formed by fibrous tissues' intertex-ture. Its' porosity was about 80%and its aperture was 100-200μm. Meanwhile, the biosecurity and biocompatibility of SIS were also fine. In the trial that the SMSCs and SIS were co-cultured in vitro, the SMSCs can grow, adhere to and differentiate on the sur-face of SIS and into the hollows very well. It can also secrete extracellular matrix. Through scanning microscope observation, cells contact with each other by their neuritis, or adhered to the wall of hole by cellular protrution. On the surface of SIS, the SMSCs maintain the property that it can easily differentiate into the chondrogenic lineage in the chondrogenic medium. Immunochemistry staining showed positive expression of type II collagen post 21 days of co-cultrue. Conclusion SIS can be used as the scaffold to construct tissue engineering meniscus as it has good biosecurity and biocompatibility with SMSCs without disturbing the cell form or inhibiting the growth and function of SMSCs.
9.Options for preventing postlaminectomy adhesion: a comparative study in rabbit model.
Chinese Journal of Traumatology 2002;5(5):288-293
OBJECTIVETo compare the various methods for prevention of peridural adhesion.
METHODSLaminectomy was performed in lumbar 1, 3 and 5 segments in 30 adult rabbits that were divided into 5 groups. A 10 mmx5 mm dura was exposed, and then covered with autogenou purifieds pearl fat (APPF) mixed with basic fibroblast growth factor (bFGF) in Group A, with APPF treated with insulin in Group B, with APPF in Group C, with 2% sodium hyaluronate (SHA) in Group D, and uncovered in Group E. The slide sections for histological study were observed at 2, 4, 6, 8 and 12 weeks postoperatively. The specimens were observed with a computed imaging analysis system at 4 and 8 weeks postoperatively.
RESULTSSevere peridural adhesions were formed in Groups B, C, D and E after laminectomy. But no adhesion was formed in the Group A at all time points. bFGF could stimulate pearl fat revascularization, increase the number of newly formed vessels and contribute to the survival of pearl fat. However, insulin had no certain effect in preventing degradation and deterioration of pearl fat. SHA had some effect in prevention of peridural adhesions only in the early stage, which was weakened with the lapse of time.
CONCLUSIONSAPPF implantation mixed with bFGF provides a potential new approach to improving fat survival and preventing peridural adhesion postlaminectomy.
Adipose Tissue ; transplantation ; Animals ; Cicatrix ; prevention & control ; Dissection ; Female ; Fibroblast Growth Factor 2 ; therapeutic use ; Hyaluronic Acid ; Laminectomy ; Male ; Models, Animal ; Rabbits ; Tissue Adhesions ; prevention & control
10.Preparation and antithrombogenicity of oxidated low molecular weight heparin-antithrombin complex coated-polyvinyl chloride tubing.
Peng LUO ; Weiyong LIU ; Chun YANG ; Hua ZHOU ; Ruijun CAO ; Jian YANG
Journal of Biomedical Engineering 2011;28(1):90-94
Based on non-enzymatic protein glycated reaction, the sodium periodate-oxidated low molecular weight heparin-antithrombin covalent complex (SPLMWATH) was produced. By using polyethyleneimine-glutaraldehyde bonding technique, polyvinyl chloride (PVC) tubings were coated with SPLMWATH, heparin and low molecular weight heparin (LMWH). Spectrophotometry and dynamic clotting time experiment were used to determine the synthetic ratio of SPLMWATH, graft density, coating leaching ratio and to evaluate the antithrombogenicity of different coating on the PVC tubings. The results showed that the synthetic ratio of SPLMWATH was approximately 55%, and compared with heparin coating and LMWH coating, the graft density of SPLMWATH coating on the PVC tubing was smaller, but its coating stability and antithrombogenicity were significantly better than that of heparin coating and LMWH coating on the PVC tubings.
Anticoagulants
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chemical synthesis
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pharmacology
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Coated Materials, Biocompatible
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chemical synthesis
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pharmacology
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Extracorporeal Circulation
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instrumentation
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Heparin, Low-Molecular-Weight
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chemistry
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pharmacology
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Humans
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Polyvinyl Chloride
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chemistry
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Surface Properties