Objective To elucidate the mechanism of inflammation in vascular endothelial cells induced by hypochlorite-modified albumin (HOCl-Alb). Methods HOCl-Alb-induced NADPH oxidase activity in human umbilical vein endothelial cells (HUVEC) was measured by lucigenin-enhanced chemiluminescence. Phosphorylation of p47phox and binding of p47phox and p22phox were measured with immunoprecipitation and Western blotting. Membrane translocation of p47phox was measured with immunofluorescence. RT-PCR and Western blotting were used to determine the intercellular adhesion molecule-1 (ICAM-1) mRNA and protein expression in the presence or absence of apocynin, respectively. Results Co-incubation of HUVEC with HOClAlb resulted in the enhancement of NAIDPH oxidase activity in time- and dose-dependent manner.Compared with bovine serum albumin group, exposure of the cells with 200 mg/L HOCl-Alb for 15min resulted in a 6.16-fold increase in NADPH oxidase activity. Phosphorylation and membrane translocation of p47phox and binding of it with p22phox were also induced by HOCl-Alb. ICAM-1expression was up-regulated after exposure to HOCl-Alb and this effect was significantly abolished by apocynin, a specific inhibiter of NADPH oxidase, in dose-dependent manner. Preincubation of the cells with 500 μmol/L apocynin inhibited the expression of ICAM-1 protein induced by HOClAlb by 68.97% (P＜0.01). Conclusion NADPH oxidase plays a central role in HOCl-Albmediated ICAM-1 expression and provides a mechanism for HOCl-Alb-related vascular endothelial inflammation.

Objective To evaluate the therapeutic effect of collagenase chemolysis after 1,2,3 years and analyse the relationship between the therapeutic effect and image features. Probing the image features for the indication, 1 254 patients with integral data who received collagenase chemolysis procedure were followed. Methods According to standard unification diagnostic criteria and therapeutic method, 1 254 patients with lumbar intervertebral disc herniation who had intergral data and received collagenase chemolysis from January 1996 to January 1999 in our department were followed up (by visit, letter or telephone call etc). The therapeutic effects of collagenase chemolysis after 1,2,3 years were evaluated with unified therapeutic effect criteria. Results For the simple lumbar intervertebral disc herniation, 1 year effective rate was 93.1%, 2 year effective rate 90.3% and 3 year effective rate 90.9%. Conclusions The simple lumbar intervertebral disc herniation is the absolute indication of collagenase chemolysis, but those combined with other factors dosen't adapt to collagenase chemolysis.

Aim To study the protective effects of SEP on the hypertrophic myocardial cells induced with high glucose and high insulin and the mechanism. Methods The protein content was assayed with Lowrys meth-od;the cardiomyocytes area was measured by computer photograph analysis system;the expression of ANF and PPAR-α was determined by RT-PCR;APS was select-ed as control drug. Results Compared with conctrol group,the protein content,cardiomyocytes area and the expression of ANF and PPAR-α of high glucose and high insulin group were significantly increased. Com-pared with conctrol group,the SEP of different dosages were able to decrease the protein content, area, the expression of ANF mRNA and PPAR-α mRNA of cul-tured hypertrophic myocadial cells induced with high glucose and high insulin. Conclusion SEP can pre-vent cardiomyocyte hypertrophy induced by high glu-cose and high insulin, which is related to its inhibition on PPAR-α signaling path.

Objective To demonstrate the inhibitory effect of adenosine A1 receptor agonist R(-)-N6-(2-phenylisopropy1)adenosine(R-PIA)and cross-talk between adenosine A1 receptor and CaN in cultured myocardial cells hypertrophy induced by Ang Ⅱ.Methods The protein synthetic rate was determined by measuring the incorporation of [3H]-leucine into myocyte protein;the cardiomyocytes volumes were measured by computer photograph analysis system;the expression of atrial natriuretic polypeptide(ANP)and CaN were determined by Western-blot.Results R-PIA(1 ?mol/L)inhibited Ang Ⅱ(0.1 ?mol/L)-induced increase of [3H]-leucine incorporation [R-PIA:(976.2?89.0)vs Ang Ⅱ:(1130.7?102.7)count per min per well,P

Aim To evaluate the effects of pioglitazone,a thiazolidinedione on the heart of diabetic animals.Methods Streptozotocin(STZ)-induced diabetic rats were divided into two groups: untreated-DM and treated-DM. Pioglitazone(20 mg?kg~(-1)?d~(-1)) was mixed in water fed to the diabetic treated group.After 5 wk,the left ventricle mass index(LVI),cardiomyocytes size and diameters,the content of hydroxyproline and collagen volume fraction(CVF) were measured.The left myocardium sections were made for HE,MASSON straining to observe the level of myocardial cells' hypertrophy and the accumulation of collagen individually and were examined by light microscope.Myocardial ultrastructure was studied by electronic microscope.Results Compared with untreated-DM group,the LVI,cardiomyocytes size and diameters,the content of hydroxyproline and CVF were lower in treated-DM group(P

AIM To investigate the effects and mechanism of cardiomyocyte hypertrophy induced by urocortin. METHODS The cardiomyocytes were divided into 8 groups: normal control, urocortin, staurosporine(Sta), verapamil(Ver), H89, urocortin+Sta, urocortin+Ver, and urocortin+H89 groups. The cardiomyocytes diameter was measured by computer photograph analysis system. The protein synthetic rate was obtained through measuring the incorporation of [~3H]-leucine into myocyte protein by liquid scintillation method. The total protein content was assayed by Lowry method. The expression of atrial natriuretic peptide (ANP) was determined by Western blot. [Ca~(2+)]_i transient was measured by Till image system by cell loading Fura-2/AM. RESULTSUrocortin group enhanced cardiomyocyte volume, protein synthesis, total protein content and expression of ANP by 30.9%, 36.3%, 35.5% and 34.7%;urocortin+Sta group decreased cardiomyocyte diameter, protein synthesis, total protein content and expression of ANP by 16.5%, 22.1%, 18.1% and 21.3%;urocortin+H89 group decreased the cardiomyocyte diameter, the protein synthesis,total protein content and expression of ANP by 16.6%, 21.5%, 19.5% and 20.6%;urocortin+Ver decreased the cardiomyocyte diameter, the protein synthesis, total protein content and the expression of ANP by 17.1%, 20.9%, 17.9% and 19.9%;Sta, H89 and Ver could decrease the [Ca~(2+)]_i transient induced by urocortin. CONCLUSION The hypertrophic effect of urocortin in rat neonatal cardiomyocytes is mediated via activation of protein kinase C and protein kinase A pathway and L-type calcium channels.

BACKGROUND: Rapid development of tissue engineering is the new hope of repairing bone defect so as to obtain complete bone regeneration,and rapid proliferation of seed cells is one of the important factors in it.OBJECTIVE: To evaluate the proliferation and osteogenic differentiation of human mesenchymal stem cells (MSCs) plated at different cell density during subculturing period.DESIGN: Repetitive measurement.PARTICIPANTS: The experiment was conducted in the Trauma, Burn and Combined Injury Laboratory of the Third Military Medical University of Chinese PLA between January 2003 and March 2004. MSCs were obtained from iliac bone bone marrow of the 5 healthy male volunteers by separation and proliferation.METHODS: The MSCs of second generation were inoculated at 8×103/cm2,3×103/cm2, and 8×102/cm2, respectively, and then the growth curve was drawn and analyzed. After 18 days of proliferation and culture, the increased amount of MSCs was recorded to analyze the post-proliferation growth curve and osteogenic ability.MAIN OUTCOME MEASURES: ①Growth curve of cells plated at different cell density; ②Results of alkaline phosphatase(ALP) staining and immunohistochemical staining of osteocalcin.RESULTS: Human MSCs from bone marrow were CD34 negative and expressed low level of ALP. Population doubling time was 40 hours after MSCs were plated at 8×103/cm2 and the number of the cells was expanded (51±13) times after 18 days. MSCs were expanded (28±6) times 18 days after plated at 3×103/cm2. While plated at 8×102/cm2, the number of the cells was only expanded (5±3) times. Proliferative ability of cells cultured at a low density was stronger that that at a high density. MSCs could be induced to differentiate into the osteocytic lineages after plated at not only 8×103/cm2 but also 8×102/cm2.CONCLUSION: Cell density plays an evident role in the proliferation of MSCs. Cell density of 8×103/cm2 is proper to the rapid proliferation of MSCs as seed cells in bone tissue engineering.

BACKGROUND:The anesthesia process is required strict in establishing rabbit models of ileal neobladder. It is required for cleaning operation field, quick postoperative recovery, and the choice of anesthetics and anesthesia does not influence smooth muscle peristalsis. OBJECTIVE:To compare the anesthetic effects between urethane and chloral hydrate in rabbits with ileal neobladder. METHODS:Eighteen rabbits were randomly assigned to urethane and chloral hydrate groups, and were respectively anesthetized with 20% urethane 5 mL/kg and 10% chloral hydrate 2 mL/kg through the ear vein. Double ureter was dissociated at 1 cm outside original bladder, and 15 cm-intestine obtained from 10 cm of ileocecal junction was implanted so as to establish models of ileal neobladder. RESULTS AND CONCLUSION:Heart rates were significantly lower compared with these before anesthesia in al cases (P≤ 0.05), but no differences were detected between two groups (P > 0.05). Compared with the chloral hydrate group, the maintain time and recovery time were longer and the mortality rate was higher in the urethane group. Above data indicated that chloral hydrate is a safe long-acting anesthetic in establishing rabbit models of ileal neobladder.

Objective To study the effect and mechanism of chlorogenic acid in allergic rhinitis(AR) mice.Methods The model of AR mice was induced by intraperitoneal injection of adjuvant suspension on every other day for 7 times, and then by nasal drip of 10% OVA for 10 days.After administration with 50, 100, 200 mg/kg of chlorogenic acid for 30 min, AR mice were attacked locally for 10 days.Behavior analysis were observed and the effect of chlorogenic acid on the contents of cytokines, IgE and histamine by ELISA in AR mice were determined.Results Chlorogenic acid decreased the number of nose-scratching events in 10min, the weight of nosal draingage, the index of spleen, the content of IL-4 in nasal lavage fluid ( NLF) and the contents of cytokines, IgE and histamine in serum; while increased the index of thymus, the content of IFN-γin NLF, the contents of IFN-γin serum and the rate of IFN-γ/IL-4.The data showed a statistically significant up-regulation of IFN-γmRNA expressions and down-regulation of IL-4 mRNA, IL-5 mRNA, IL-10 mRNA expressions after adiminstration of chlorogenic acid.Conclusion Chlorogenic acid might exert the effect in AR mice through regulating the balance of Th1 and Th2.