Bone marrow-derived stromal cells and cartilage tissue engineering

Articular cartilage repair is one of the most challenging issues which remains to be resolved in clinic work. Discovering of bone marrow stromal cells (BMSC) and its application in tissue engineering provide new methods for the treatment of cartilage defects. High seeding density, appropriate cytokines and three-dimensional culture play important roles in the process of inducing BMSC differentiating into chondrocytes, suitable scaffold is also essential in reconstructing cartilage in vitro by methods of tissue engineering.     

Effect of cocaine on caspase-3 in myocardiac cells of male rats in different age

AIM: To study the effect of cocaine on caspase-3 in myocardiac cells of male rats in different age. METHODS: Three-week-old(n=16), six-week-old(n=16) and twelve-week-old (n=16) male Sprague-Dawley rats were all divided into control groups and experiment groups randomly, each group had eight animals, experiment groups were given cocaine hydrochloride (15 mg·kg^-1 body weight) subcutaneously daily for four weeks. The ratio of heart weight to body weight (HW/BW, mg/g) were measured. DNA fragmentation of myocardia cells was determined by gel electrophoresis, and caspase-3 activity in myocardia cells was tested by flow cytometry (FCM). RESULTS: In three experiment groups, the DNA isolated from myocardial cells displayed clear ladder pattern. The HW/BW and the caspase-3 activity were increased significantly than those of control groups (P<0.01). CONCLUSIONS: Cocaine induced apoptosis in rat myocardial cells. The increase in caspase-3 activity may be the one of important pathways related to the induction of apoptosis.     

Effects of hypoxia and hypercapnia on expression of soluble guanylate cyclase in rat pulmonary artery

AIM:To investigate the expression of soluble guanylate cyclase protein and its mRNA in rat pulmonary artery after exposure to hypoxia and hypercapnia.METHODS:Male Sprague-Dawley rats were randomly split into 4 group, which were hypoxic hypercapnic (HH 1 week, HH 2 weeks, HH 4 weeks) group and control group, to copy pulmonary hypertensive animal model. The expression of sGCα₁ and β₁ subunits protein of medial and small pulmonary artery was performed by immunohistochemistry with a polycolonal antibody. In situ hybridization was performed on the rat lung tissue using sGC oligonuclear probe to assay the expression of sGCα₁subunit mRNA.RESULTS:The sGCα₁ and β₁ subunits protein and sGCα₁ subunit mRNA were faint staining in the pulmonary small and medium artery in HH1 week, HH 2 weeks and HH 4 weeks groups compared with control group (all P＜0.01).CONCLUSION:sGC subunit mRNA and protein expression in pulmonary small and medium artery were decreased after exposure to hypoxia and hypercapnia, which took part in the development of the pulmonary hypertension.     

Rat urotensin-II-induced main pulmonary artery contractions are mediated by MAPK

AIM: To investigate rat Urotensin-II(rat U-II)-induced vasoconstriction of rat main pulmonary arteries and the role of mitogen-activated protein kinase(MAPK). METHODS: The main pulmonary artery was dissected from the male Sprague-Dawley rats and artery ring width was 3-4 mm. Concentration-response curves were generated to rat U-II(0.03 nmol/L-30 nmol/L).Inhibitor of MAPK, PD 98059(0.1 μmol/L-10 μmol/L) were added into the medium after rat U-II(30 nmol/L)induced vasoconstriction had reached plateau to construct the relaxant concentration-response curves and their EC₅₀ and E_max. RESULTS：Rat U-II was a potent vasoconstrictor of isolated rat main pulmonary arteries [EC₅₀=7.95±0.40, E_max=(14.28±6.34)% of the response to 60 mmol/L KCl]; PD 98059 caused concentration-dependent relaxations of rat U-II precontracted arteries [EC₅₀=5.91±0.45, E_max=(81.39±13.65)%]. CONCLUSION: Rat U-II was a potent vasoconstrictor of rat main pulmonary arteries and this response was mediated through MAPK.     

Effects of FK506 on anti-glomerular basement membrane nephritis in rats

AIM:To investigate the effects of FK506 on anti-glomerular basement membrane(GBM) nephritis in rats. METHODS: Anti-GBM nephritis model was elaborated by rabbit anti-rat GBM antibody injection in SD rats in this study. The rats were divided into three groups: FK506 treated group(0.5 mg·kg^-1·d^-1, sc), untreated nephritis control group and normal control group. FK506 was administered daily six hours after injection of anti-GBM IgG. All the rats were observed urinary protein at the 4th day, the 14th day and the 21st day. At the same time, the kidney specimens were collected, and T cell transforming function was also monitored.RESULTS:Rats injected with rabbit anti-GBM Ab developed heavy proteinuria by 4 days, and serum creatinine and serum urea appeared which kept on the rising. Glmerular hypercellularity, crescents, and protein casts were observed in nephritic rats. By electron microscopy, the thickening of GBM and loss of foot processes were seen. T cell transforming function was higher than normal. But, all pathological changes obviously turned for the better in FK506 treated group. CONCLUSION:FK506 could inhibit the progression of rat anti-GBM nephritis.     

Therapeutic effects of recombinant human growth hormone on rat sepsis

AIM:To investigate therapeutic effects of recombinant human growth hormone(rhGH) on rat sepsis and its possible mechanisms.METHODS:Mean arterial pressure (MAP), levels of plasma TNFα, IL-1β and endotoxin, leukocyte count and survival rate within 1 week were determined after E. coli injection among control group, sepsis group and sepsis+rhGH group.RESULTS:(1)rhGH diminished the decrease of MAP, reduced plasma endotoxin and TNFα levels and increased neutrophil ratio in total leukocytes in sepsis rat. rhGH increased survival rate within 1 week on sepsis rat. (2)No changes were found in IL-1β level among the three groups.CONCLUSION:rhGH showed desirable beneficial effects on rat sepsis, which may attribute to: improving circulatory function;maintaining intestinal mucosa barrier, attenuating bacteria/endotoxin translocation and inhibiting the production and release of TNFα.     

Serum contents of E-selectin,sICAM-1, sVCAM-1 in patients with acute coronary syndrome

AIM:To explore the serum levels of certain adhesion molecules and its significance in acute coronary syndrome(ACS). METHODS:The subjects included 40 patients with acute myocardial infarction(AMI) and 40 patients with unstable angina pectoris (UAP). Among the 80 patients, 60 patients accepted a follow-up 4 months. At the same time we selected 40 controls from people who attended a routine health check in the university. Serum levels of E-selectin, sICAM-1, sVCAM-1 were measured by ELISA.RESULTS:Serum levels of E-selectin, sICAM-1, sVCAM-1 were significantly higher in the ACS group(AMI or UAP) than in the control group. Four months later, the levels of E-selectin, sICAM-1 became significantly lower in the follow-up group than in the ACS group, while sVCAM-1 showed no significant difference. CONCLUSION:Serum levels of E-selectin, sICAM-1 may have certain diagnostic value for ACS, and can be a useful marker reflecting the stability of the disease.     

Effect of tea-polyphenols on the activation of NF-κB and the expression of TGF-β1 mRNA in THP-1 cells incubated with VLDL,LDL or ox-LDL

AIM:To investigate the effect of tea-polyphenols (TP) on the activation of NF-κB and the expression of TGF-β₁ mRNA in THP-1 cells (a human acute monocytic leukemia cell line). METHODS:THP-1 cells were incubated with the different concentrations of TP, VLDL, LDL or ox-LDL. In the THP-1 cellls, the nuclear malposition rate of NF-κB was detected with immunohistochemistry technique, the positive index of the TGF-β₁ mRNA expression was detected by hybridization in situ, and accumulation of total cholesterol (TC) in cells incubated with 0.4-40 μg/L TP was determined with oxidase assay. RESULTS:The nuclear malposition rate of NF-κB, the positive index of the TGF-β₁ mRNA expression and TC in THP-1 cells incubated with 0.4-40 μg/L of TP were lower than those with 0 μg/L of TP in TP-V group, TP-L group and TP-O (P＜0.05). The differences of these markers in THP-1 cells incubated with more than 40 μg/L TP in TP-V group, TP-L group and TP-O were not statistically significant, compared with TP-C group (P＞0.05). CONCLUSION:TP inhibited the activation of NF-κB, the expression of TGF-β₁ mRNA and the foam cell formation in the mono-macrophage.     

Effect of fibronectin on proliferation and collagen synthesis in cardiac fibroblasts from spontaneously hypertensive rats

AIM:To investigate the effects of fibronectin (FN) on the proliferation and collagen synthesis in cultured rat cardiac fibroblasts (CFb) derived from SHR (CFb_SHR) and WKY (CFb_WKY). METHODS:CFb derived from 12-week-old spontaneously hypertensive rat (SHR) and WKY was cultured by outgrowth of tissue block. Cell proliferation of CFb was measured by cell number counting and[³H]-TdR incorporation using 24-well plates pre-coated with 5 μg/cm² of FN. Collagen synthesis was determined by [³H]-proline incorporation. RESULTS:As compared with control, the cell number of fibroblasts derived from SHR and WKY were significantly increased to 163.75% and 170.42% respectively after 72 h incubation with FN in the presence of 0.4% FCS from a intial cell density of 1×10⁴ cells/mL. DNA synthesis of CFb was markedly promoted by FN. FN induced an increased in [³H]-proline incorporation in both CFb_SHR and CFb_WKY. CONCLUSION:FN is able to promote cell proliferation and collagen synthesis of CFb derived both from SHR and WKY.     

Effect of cilazapril on pulmonary vascular endothelial dysfunction in hypoxic rats

AIM:To study the effect of cilazapril on pulmonary vascular endothelial dysfunction in hypoxic rats. METHODS:The structure and function of endothelium in hypoxic rats were studied by biochemical analysis, radioimmunoassay, transmission electron microscope and correlated with hemodynamic. RESULTS:1) The change and damage of ultrastructure in endothelial cell (EC) were obsevered in hypoxic rats. 2) The contents of plasma nitric oxide (NO) and superoxide dismutase (SOD) activity in blood as well as endothelial nitric oxide synthase (eNOS) activity in the lung tissue were significantly lower in the hypoxic rat than those in contral animals. The concentrations of plasma endothelin-1(ET-1) and angiotensin converting enzyme(ACE) as well as malondialdehyde(MDA) were significantly higher in the hypoxic rat than these in contral animals. The relaxing and contracting factors had a significant positive/negative correlation with mean pulmonary artery pressure (mPAP). 3) Cilazapril significantly decreased the level of ET-1 and ACE and significantly increased the level of NO and activity of eNOS and SOD. At the same time, cilazapril extenuated hypoxia-induced injuries of EC. CONCLUSION:The results indicate that damaging structure and dysfunction of EC existes in hypoxic rats. The cilazapril effectively preventes and treates the chronic hypoxic PH by relieving the injury and improving secretion in EC.