Abstract:Objective:To investigate the alteration of cell proliferation and tazarotene-induced gene 3(TIG3) mRNA level in cultured normal human keratinocytes after tazarotene and narrow-band UVB(NB-UVB) combined treatment or alone. Methods:Normal human keratinocytes were exposed to NB-UVB(50-100 mJ/cm2) and/or followed incubation with tazarotene(0.1-1 μmol/L) for 24 hours,then the proliferation potency of the cells was detected by methyl thiazoleterazolium colorimetric assay and the level of TIG3 mRNA was examined by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR). Results:Cultured keratinocytes proliferation was inhibited and TIG3 mRNA level was enhanced by both two doses of tazarotene single treatment but not by NB-UVB irradiation alone. Further,the higher dose of tazarotene showed more effects on inhibiting keratinocyte proliferation and elevating TIG3 mRNA level. Tazarotene-NB-UVB combination showed great effects on inhibiting keratinocyte proliferation and increasing TIG3 mRNA level,which were stronger than those induced separately. Conclusion:Tazarotene incubation single can inhibit keratinocyte proliferation and enhance TIG3 mRNA level,but NB-UVB irradiation alone can’t. Tazarotene-NB-UVB combination showed synergistic effects on inhibiting keratinocyte proliferation and increasing TIG3 mRNA level.

Abstract:Objective:To investigate whether neuronal nitric oxide synthase(nNOS) neurons in the hypothalamic paraventricular nucleus(PVN) and supraoptic nucleus(SON) were involved the central pathways of the arterial baroreflex in rats. Methods: Immunocytochemical method for Fos combining with nicotinamide adenine dinucleotide phosphate diaphorase(NADPH-d) histochemical technique for nNOS was used to localize the Fos and NADPH expressions in the subdivisions of the PVN and SON following sinoaortic baroreceptor denervation(SAD). Results:It was found that distinct number of Fos positive neurons coexisting with NADPH-d in the parvocellular subdivisions, magnocelluar subdivisions of the PVN and the whole SON. The percent of Fos/NADPH-d double-labeled neurons in the above regions was 6.8%, 72.1% and 47.1% respectively. Only few Fos positive neurons were observed at the above regions in the sham-operated and intact control rats, and no NADPH-d/Fos double-labeled neurons were detected. Conclusion:The nNOS neurons in the PVN and SON are activated by the SAD specifically, which highly indicates they may take part in the central cardiovascular regulation mediated by the arterial baroreflex.

Abstract:Objective:To investigate the efficiency of transducing rat bone marrow mesenchymal stem cells(BMSCs) with enhanced green fluorescent proteins(eGFP) gene in vitro by a chimeric adenovirus serotype 5 vector Ad5/F35 which carries an adenovirus serotype 35 fiber and observe the longevity of transgene expression in transduced rat BMSCs. Methods:Rat BMSCs was isolated and proliferated by adhesive culture; transduced rat BMSCs of passage 2 with eGFP gene using Ad5/F35 carrying an eGFP gene(Ad5/F35-eGFP) at a multiplicity of infection(MOI) of 1,10,100 and 1000 respectively; tested the transduction efficiency and expression level of eGFP gene in rat BMSCs by fluorescent inverted microscope and flow cytometer(FCM); studied the basic biological characteristic of transduced rat BMSCs by observing their growing state; tested the eGFP expression in rat BMSCs at day 2,7,14,21 and 30 respectively after transduction at a MOI of 100 by FCM. Results:A positive dose-response relationship was observed between the transduction efficiency and expression of eGFP gene and the Ad5/F35-eGFP concentrations ranging from 1~1000 MOI. Approximately 90% of the rat BMSCs can be transduced by Ad5/F35-eGFP at a MOI of 100,and the transgene expression can be detected for a month. The virus which infects rat BMSCs at a MOI of 1,10 and 100 can not impair the vitality and proliferative capability of transduced rat BMSCs. Conclusion:Ad5/F35-eGFP can transduce rat BMSCs with eGFP gene efficiently in vitro,the transgene expression in rat BMSCs can last a month. These data suggest that the Ad5/F35 is an effective vector of BMSCs for gene transfer and expression and it will be a powerful tool for gene therapy and cell therapy based on BMSCs.

Abstract:Objective:To study the effect of verapamil on platelet aggregation, thrombogenesis and the potential mechanism. Methods:The SD rats were injected the different concentration of verapamil, Aspirin-DL-Lysine and the saline solution for 7 days(i.p), respectively. After the administration, the platelet aggregation was measured by aggregometry, the thrombogenesis was tested by the pathway of arteria carotis communis-external jugular vein, and the concentration of NO was evaluated with the kit of nitrate reductase. Results:Verapamil exhibited a dose-dependent inhibitory effect on platelet aggregation and thrombus formation in rats. Conclusion: Verapamil imporoving the function of platelet was through elevating the NO level.

Abstract:Objective:To develop and validate a three dimension finite element model of the earlier-period degeneration L4-5 lumbar motion segments reconstruced from CT Images. Method:To develop such FE model,a 138 frames of CT images with the distance 0.75 mm were gathered in clinic,from which the plane coordinate data were obtained,and then these 2D data were transformed into 3D coordinate. Finally,the 3D FE model of a Chinese woman the earlier-period degeneration L4-5 lumbar motion segment was developed using Patran analysis software,and then the model was validate by a 400N stress force. Results:The result shows that the model has good effect visually and mathematically,which was consisted of 27 130 nodes,113 834 elements,including Solid elements 113 153,Area elements 161,Link elements 520. The results of load test of the aforementioned FE model was consistented the published results. Conclusion:Based on CT image and with the help of CAD methods,this three dimension finite element model of the earlier-period degeneration L4-5 lumbar motion segments can be constructed automatically with excellent simulation results.

Abstract:Objective:To cultivate rat MSCs and explore the protective effects of MSCs on the transplanted kidney. Methods:Bone marrow was collected from SD rat by flushing femurs and tibias under sterile condition. MSCs were purified by stillness-stick cultivation in vitro. Forty wistar rats were transplanted with the left kidneys of SD rats,twenty wistar rats were infused with MSCs(Group I),not did the other twenty Wister rats(GroupⅡ). At day3,the right renal pedicle of recipients were ligated. The survival time and morphology of the transplanted kidneys were observed. Results:The survival time of Group I(15.50 ± 6.35)d was longer than that of Group Ⅱ[(6.50 ± 3.11)d,P < 0.05]. The former transplanted kidney morphology observation and function obviously surpass the latter. Conclusion:MSCs can be isolated,cultured and expanded easily in vitro,and have protective effect on the transplanted kidney in early period.

Abstract:Objective:In order to know the clinical significance of the CD8+CD25+ T cell in autoimmune disease,hypersensitivity disease and organ transplantation, the normal limits of this T cell subset was obtained by studying the number of CD8+CD25+ T cell and its productions of IFN-γ,IL-10 and TGF-β in the normal peripheral blood. Methods:Mononuclear cells were isolated from healthy donors. The percentages and absolute numbers of CD8+CD25+,CD8+CD25-,CD8-CD25+ and CD8-CD25- from CD3+ T cells were detected by flow cytometry. In addition,stimulating with/without ionomycin and PMA 5 hours later,the expressions of IFN-γ,IL-10 and TGF-β in the CD8+CD25+ T cells was measured. Results:The percentage of CD8+CD25+ subgroup in CD3+ T cells from normal human peripheral blood was 2.52%,the absolute number was 37.09×106/L. The percentage and absolute number decreased after stimulated with ionomycin and PMA 5 hours. The expression of TGF-β on the surface of CD8+CD25+ T cells up to 93.8%,stimulation seems not obviously affected its expression but IFN-γ+ T cells increased obviously,however,IL-10+ T cells didn’t obviously up-regulat. Conclusion:There were a few CD8+CD25+ T cells in CD3+ T cells from normal peripheral blood. They expressed highly TGF-β in the cell surface,and could have IFN-γ reaction but no proliferation after stimulation.

Abstract:Objective:To investigate the effect of Valsartan on the expressions of Toll-like receptor(TLR) 2 in the arteria of streptozotocin(STZ) induced diabetic rats. Methods:Thirty-one Sprague-Dawley rats aged 8 weeks were assigned randomly to receive STZ at a dose of 60 mg/kg intraperitoneally(diabetes, n = 24) or citrate buffer alone(normal controls,NC,n = 7). One week after injection, only STZ-treated rats with plasma glucose concentrations above 16.7 mmol/L were considered diabetic and were subsequently dividied into two groups:diabetic group and DV group which received Valsartan 20 mg/(kg·d) intragastricly. At 12th week, the rats in three groups were sacrificed for their arteria and blood samples. The expressions of TLR2 mRNA were determined by reverse transcription PCR. The protein levels of TLR2 were detected by Western blot. Results:Compared with NC group, the levels of blood glucose in DM and DV groups increased significantly(P < 0.01), but no notable difference was found in the levels of blood glucose between DM and DV group(P > 0.05). The expressions of TLR2 mRNA and protein in DM group were much higher than that in NC group(P < 0.01), but it was much lower in DV group than that in DM group(P < 0.01). Conclusion:The expressions of TLR2 mRNA and protein were much higher in the arteria of diabetic rats, Valsartan could obviously reduce the expression of TLR2 in the arteria of diabetic rats which implicates that it might be able to ameliorate the early inflammatory reaction in the arteria of the rats with diabetes.

Abstract:Objective:To investigate the relationship between the expression of nNOS-HO-2 and the high contration glucose treatment in the interstitial cells of cajal(ICC) in vitro. Methods:The ICC was isolated from the proxinal colon in newborn rats,and divided into control group,high glucose group and mannitol(osmolarity control) group. Cells viability was assessed by the MTT assay,and the morphologic changes of ICC was observed under microscopy. The expression of nNos and HO-2 was evaluated by Western-blotting. Results:Compared to the controls,high glucose markedly decrease the cell viability. After treatment of high glucose,most of the ICC lost their network of each other,and the expression of nNOS and HO-2 was also decreased. In mannitol treatment ICC,there was no changes in cell viability,cell morphology and nNos and HO-2 compared to the control cells. Conclusion:High glucose treatment could induce ICC cell damage and inhibit the expression of nNos and HO-2. The abnormal expression of nNos and HO-2 may play an important role in the intestines dysfunction of diabetic mellitus.

Abstract:Objective:To investigate the effect of tissue inhibitor of metalloproteinase-1(TIMP-1) on the apoptosis of β cell induced by hydrogen peroxide and the mechanism of TIMP-1 action. Methods:The insulin-producing rat cell line RINm5F cells were cultured and stimulated by H2O2 with or without the addition of TIMP-1(50 μg/L). Cell viability,Caspase-3 and superoxide dismutase(SOD) activity were determined after the cells were exposed to H2O2 and TIMP-1. Results:Cell viability was decreased obviously in RINm5F cells after incubation with H2O2(0.1 mmol/L) for 1 hour alone. In the cells after treatment with TIMP-1(50 μg/L) for 1 hour prior to H2O2,cell viability and SOD activity were remarkably increased and the activity of Caspase-3 was lower significantly than that in the cells treated by H2O2 alone(P < 0.05). Conclusion:TIMP-1 can protect RINm5F cells from apoptosis induced by hydrogen peroxide,which possibly involves in raising the activity of SOD.

Abstract:Objective:To establish the nonobese diabetic-severe combined immune deficient(NOD/SCID) mice model inoculated with human leukemia cell line K562 cells,which encoded a marker gene-green fluorescence protein. Methods:GFP-encoded K562 cells were obtained by the infection of K562 cells with recombinant retrovirus encoded GFP. After total body irradiation with γ-ray of 2.5Gy,NOD/SCID mice were intravenously transplanted with 5 × 105 K562-GFP cells(Group 1,n = 3),or 1 × 106 K562-GFP cells (Group 2,n = 3),or normal saline(Control,n = 2). Six weeks after transplantation,the bone marrow,peripheral blood and spleen of recipient mice were assayed for the presence of human cells by flow cytometry and PCR. Results:Six weeks posttransplatation,the presence of human leukemia cells was found in the peripheral blood,bone marrow and spleen of recipient mice. The mean percentages of human cells in the bone marrow of recipient mice of Group 1 and Group 2 were 12.3% and 22.6%,respectively. Conclusion:human leukemia model inoculated with GFP-encoded k562 cells was successfully established,which could be employed in the future research.

Abstract:Objective:To investigate the effects of ketamine on production of Nitric oxide(NO),hydroxy radical(·OH) and superoxide anion(O2-·) in the rat alveolar macrophage cell line(NR8383) activated by Lipopolysaccharide(LPS). Methods:Macrophage was distributed into five groups,group C,group L,group K1,group K2 and group K3. Group L,K1,K2 and K3 were stimulated with LPS. Before stimulation,Group L,K1,K2 and K3 were treated with phosphate buffer solution(PBS),10,100 and 1 000 μmol/L ketamine. Group C was treated with PBS. The levels of NO,·OH and O2-· in the supernatant were assayed. Results:The concentration of NO,·OH and O2-· in group L was increased significantly(P < 0.01) when compared to Group C. The concentration of NO,·OH and O2-· in Group K2,K3 was obviously lower than those of Group L(P < 0.05). There was no statistical difference between K2 and K3(P > 0.05). Conclusions:Ketamine can inhibit LPS-induced production of NO,·OH and O2-·.

Abstract:Objective:To investigate the relationship between serum IL-10 and apoptosis of T lymphocytes in systemic lupus erythematosus(SLE). Methods:ABC-ELISA was used to analyze the level of serum IL-10. Three-color flow cytometry was used to determine the changes of percentages of peripheral T lymphocyte subsets,their expressions of Fas and FasL,and of their early apoptosis. And then,the relationship between these changes and the level of serum IL-10 was analyzed. Results:In SLE,compared with normal controls,the serum IL-10 level increased significantly,and had a positive correlation with SLEDAI and the level of serum anti-dsDNA antibodies. In peripheral blood,the percentage of CD4+ cells and CD4+/CD8+ ratio was decreased,and the expression rates of Fas and FasL of T lymphocyte subsets was increased. The early apoptosis of T lymphocyte subsets,especially of CD4+ subpopulation,was increased. These results above became more obvious in SLE patients with active disease. In SLE,there was a negative correlation between serum IL-10 level and CD4+/CD8+ ratio,while a positive correlation between serum IL-10 level and the increased expression of FasL, and the increased apoptosis of CD4+ T lymphocyte. Conclusion:The abnormally increased serum IL-10 level in SLE patients can induce a high level of FasL,which mediates the apoptosis of CD4+ T lymphocyte,and involves in a more active disease.

Abstract:Objective:To explore the changes of the myelin-associated oligodendrocytic basic protein(MOBP) gene mRNA and protein expression in the prefrontal cortex(PFC) of SHR/WKY and to analyze the relationship between the MOBP and the etiology and pathophysiology of Attention Deficit Hyperactivity Disorder(ADHD). Methods:The expressive levels of MOBP in the PFC of SHR and WKY were verified by using RT-PCR technique. The distribution of the MOBP in rat brains was marked by used MOBP-HRP immunohistochemistry(IHC) and the electron microscope were applied as the ultrastructural study on brain tissues. Results:The levels of MOBP gene mRNA expression(35.78 ± 6.04)% in the PFC of SHR were significantly lower than those(69.41 ± 7.43)% in the PFC of WKY rats. At the same time,there was ultrastructural difference in arrangement of the radial component in myelin between two groups. Conclusion:The less of MOBP expression in the SHR may be one of the important pathophysiological bases for ADHD.

Abstract:Objective:To explore the role of ERK signal pathway in the extracellular matrix(ECM) synthesis of rat mesangial cells treated with PDGF-BB in vitro. Methods:Cultured rat mesangial cells were divided into seven groups:control cells; treated with PDGF-BB(10,25,50 ng/ml,respectively);treated with PDGF-BB(25 ng/ml) and UO126(5,10 -滋mol/ml,respectively);treated with UO126 10 -滋mol/ml. The expression of fibronectin(FN),matrix metalloproteinases-2(MMP-2) and plasminogen activator inhibitor-1(PAI-1) mRNAs were detected by RT-PCR. Results:In mesangial cells,PDGF-BB downregulated MMP-2 mRNA expression(P < 0.05),upregulated PAI-1and FN mRNAs expression in a dose-dependent way(P < 0.05). The changes of FN,MMP-2 and PAI-1 mRNA induced by PDGF-BB can be inhibited by UO126 in a dose-dependent fashion(P < 0.05). Conclusion:PDGF-BB can inhibit the degradation of extracellular matrix and stimulate the synthesis of FN in rat mesangial cells through ERK siginal pathway.

Abstract:Objective:To investigate the antioxiative effects of spironolactone ,an aldosterone receptor blockade,on kidney of rats with unilateral ureteral obstruction(UUO). Methods:Thirty wistar rats underwent UUO and then received vehicle(n = 11) or spironolactone(50mg·kg-1·d-1 by daily gastric gavage,n = 12) during the 15 days from the day before surgery. Additional 7 rats were sham operated. All the rats were killed at 14 days after surgery. Histological changes were observed by HE and Masson staining. Malondialdehyde(MDA),superoxide dismutase(SOD) as well as aldosterone(ALD) content were measured. Results:UUO induced a significant increase in MDA and ALD content but a marked decrease in SOD content and severe morphology changes. However,spironolactone ameliorated what had mentioned. Conclusions:Spironolactone can suppress oxidative stress and alleviate renal in rats with UUO by reducing lipid peroxidation production and improving antioxidative enzyme contents.

Abstract:Objective:To assess the diagnostic accuracy and clinical application of three-dimensional(3-D) 16-slice CT angiography(CTA) in detecting cerebral arteriovenous malformations(AVM). Methods:70 cases suspected cerebral AVM received CT scan and digital subtraction angiography(DSA),and the results were compared retrospectively. CT posterior reconstructions were carried out by using volume rendering(VR),maximum intensity projection(MIP) and shaded-surface display(SSD). Results:17 of 70 patients had cerebral AVM,24 patients had cerebral aneurysms,three patients had moyamoya disease,and 26 patients were negative. In 17 AVM patients,all patients found malformated vessel masses,16 patients clearly displayed feeding arteries and 15 patients demonstrated draining veins in 3-D 16-slice CTA images. Conclusion:16-slice CTA has a very high accuracy of detecting AVM. 16-slice CTA is sensitive enough to replace conventional DSA in the triage,diagnosis,and treatment planning in patients with cerebral AVM.

Abstract:Objective:To evaluate the correlation between the expression of Pin1 and β-catenin. To identify whether Pin1 was a prognostic factor in colorectal cancer. Methods:The expression of Pin1 and β-catenin proteins was detected by immunohistochemistry in 50 cases of colorectal cancer tissues. The relationships between Pin1 and β-catenin expression and clinicopathological factors were evaluated. Results:The overexpression of Pin1 and the aberrant accumulation of β-catenin in colorectal cancer was 36%(18/50) and 58%(29/50),respectively. A significant positive correlation was found between Pin1 and β-catenin(P = 0.006). The overexpression of Pin1 was related to pathological grade and tumor stage(P < 0.05). Pin1 was significantly related to the overall survival(P < 0.01). Patients with negative β-catenin expression survived longer than those with aberrant expression despite that no statistic significance was detected(P = 0.099). Conclusion:Pin1 was overexpressed and β-catenin was aberrant accumulated in colorectal cancer in this study. There was a significant correlation between their expression. Pin1 maybe a prognostic factor in colorectal cancer.

Abstract:Objective:To investigate retrospectively the clinical significance the serum C-reactive protein(CRP) in patients with relapse of malignant tumor before and after chemotherapy. Methods:The values of serum CRP were quantitatively determined by immunoephelometry. Thirty-four patients with relapse of malignant tumor were served as the experiment group,and 22 patients with adjuvant chemotherapy after surgical resection were served as control group. Results:CRP elevation was found in 83.25% of patients with relapse of malignant tumor. No obvious change of CRP was found in patients with adjuvant chemotherapy after surgical resection. CRP was decreased in chemotherapy responders after chemotherapy compared to before chemotherapy,but it was increased in chemotherapy nonresponders. Median survival time was 8 months in patients with ineffective chemotherapy. Conclusion:The curative effect was inverse correlation with the CRP level and the increase of CRP after chemotherapy indicates the poor outcome. Maybe CRP could be served as a reference to diagnosis,curative effect and prognosis for patients with relapse of tumor.

Abstract:Objective:To study the effect on Leptin by investigating the level of Leptin in serum of rats with brain injury and spinal injury combined femoral fracture. Methods:Seventy-two male SD rats were randomly divided into 6 groups:normal control,fracture group,brain injury group,spinal injury group,fracture with brain injury and fracture with spinal injury group(n = 16 per group). Fracture,spinal moderate injury and brain moderate injury models were established. The rats were sacrificed 2,4,8 and 12 weeks respectively after operation,and then the Leptin concentration in serum was detected by rat Leptin. Results:The serum levels of Leptin in the brain injury group,fracture group or fracture with brain injury group significantly increased as compared with that in control group at the 2nd week after operation(P < 0. 05). The concentration of the Leptin in serum of the fracture with brain injury group was higher than other operative group 4 and 8 weeks after operation(P < 0. 05); at the 12th week after operation the difference in above 6 groups was not statistically significant(P > 0. 05). Conclusion:The lever of Leptin in serum was higher in fracture with brain injury group than other groups in certain period,which suggests that Leptin probably makes a comparatively strong responsiveness to fracture with brain injury group.

Abstract:Objective:To study the isolation rate of Acinetobacter species(including Acinetobacter baumannii and Acinetobacter calcoaceticus-baumannii),survey and analyze the changes of Acinetobacter resistance to common antimicrobial agents. Methods:The bacteria were identified by API system; the sensitivity of antimicrobial agents was tested by K-B method; the data was analyzed by WHONET 5.4. Results:The isolation rate of Acinetobacter was increasing year-by-year from 3.3% in 2002 to 6.8% in 2006; and the resistance rates of Acinetobacter to common antimicrobial agents was increasing. The three of the highest increase as follows:Amikacin from 21.9% to 59.6%,Piperacillin/tazpbactam from 25.4% to 54.6%,Ceftazidime from 41.5% to 69.1%. Conclusion:It’s important that surveillance data of the resistance rates of Acinetobacter to common antimicrobial agents,place importance on high increases of the resistance rates and select antimicrobial agents rational.

Abstract:Objective:To study the association between cigarette smoking and the severity of coronary atherosclerosis. Methods:The study population consisted of 1060 consecutive patient who underwent coronary angiography for suspected or known coronary atherosclerosis. The severity of coronary atherosclerosis was defined by Gensini’s score .The Spearman two-way test and partial correlation analysis was used to assess the relation between two quantitative variables. The independent-sample T test was employed to investigate the distribution of the clinical data in two groups according to smoking status. Multivariate stepwise linear regression analysis was employed to explore the independent relationship between smoking and the severity of coronary atherosclerosis. Results:Spearman’s correlation analysis indicated that smoking was positively correlated with leukocyte count, neutrophil, CH/HDL-C, LDL-C/HDL-C and the Gensini’s score. Leukocyte count, neutrophil, CH/HDL-C and LDL-C/HDL-C was positively correlated with the Gensini＇s score, and partial correlation analysis showed that smoking was still positively correlated with the Gensini’s score while the age and the sex were under controlled. Independent-sample T test also found the significant differences in terms of leukocyte count, neutrophil, CH/HDL-C and LDL-C/HDL-C between the smoking group and non-smoking group. Multiple stepwise linear regression analysis demonstrated that neutrophil, smoking index and LDL-C/HDL-C significantly independently associated with Gensine’s score. Conclusion:Smoking was independently associated with the severity of coronary atherosclerosis, as measured by Gensini＇s score. Inflammatory response and hyperlipidemia may be the mechanism involved.