Objective:To study the influence of TNF-? on the expression of secretory component (SC) in Caco-2 cells.Methods:Immunocytochemistry,ELISA,Western blot and quantitative real-time PCR were used to test SC-positive cells of Caco-2,free SC in culture supernatants,protein and mRNA expression of SC in the cells.Results:The increase of SC-positive cells,free SC in culture supernatants,SC protein expression and expression of SC genes in Caco-2 cells was found under stimulation of TNF-? treatment compared with control(P

Objective To explore the influence of Shensongyangxin capsule on plasma high-sensitivity C-reactive protein(hs-CRP) and N-terminal pro-brain natriuretic peptide(NT-proBNP) in patients with persistent atrial fibrillation after cardioversion.Methods 152 patients with persistent atrial fibrillation were randomly divided into the observation group(76 cases)and the control group (76 cases).All patients were conducted atrial cardioversion.The patients were followed up for 12 months.The plasma hs-CRP and NT-proBNP were measured before and after treatment.Results All patients with atrial fibrillation were cardioversed to sinus rhythm after cardioversion.During 12 months of follow-up,in the observation group 2 patients had recurrence of atrial fibrillation and in the control group,nine cases of recurrence,the difference between the two groups was significant (x2 =5.28,P ＜ 0.05).After treatment,the plasma hs-CRP and NT-proBNP in the two groups were significantly decreased(t =7.270,3.601,8.118,3.006,P ＜ 0.05,P ＜ 0.01),and those in the observation group were significantly lower than control group (t =4.720,2.914,all P ＜ 0.05).Conclusion Shensongyangxin capsule can significantly reduce plasma hs-CRP and NT-proBNP in patients with persistent atrial fibrillation after cardioversion.

Objective To study the changes of IgA,secretory component(SC)and ZO-1,occludin of gut in severe hepatitis and to understand the reason of abdomen symptom in severe hepatitis patients.Methods IgA,SC,ZO-1 and occludin of gut were assayed by immunohistochemistry.Results Compared with the controls,the staining of IgA,SC,ZO-1 and occludin in severe hepatitis were notably decreased.Conclusion In severe hepatitis,IgA,SC,ZO-1 and occludin expression of gut decrease,leading to the abnormality of barrier of gut,which is one the reasons of resuhings in abdomen symptoms in severe hepatitis.

Objective To develop a radiology algorithm and test its the accuracy in distinguish pacing in the septum from the other parts. Methods One hundred patients were implanted with double-chamber pacemakers. Sites of the leads were verified by two-dimensional echocardiography, and the patients were divided into 4 groups according to the echocar?diography:septal right ventricular outflow tract group(RVOT), RVOT anterior free wall group, mid septum group, and anteri?or septum group (near to the anterior free wall ). An algorithm was developed according to radiological characteristics in the 45° left anterior oblique (LAO45° ) view and the 30° right anterior oblique (RAO30° ) view. Then its sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were tested . Results The algorithm has high sensitivi?ty and specificity, which were 90%and 97%respectively. The positive predictive value and negative predictive value were 90% and 97% respectively. Conclusion The radiology algorithm we developed have a high sensitivity and specificity in identifying the site of the leads.

BACKGROUND: Nowadays, fish collagen biomedical materials still exhibit obvious deficiency in thermal stability,in vivo degradation stability and in vivo material morphology stability. To expand the application of fish source collagen, it is urgent to improve the material performance by increasing the density and collagen molecule tightness of freshwater fish collagen sponge materials using technique methods.OBJECTIVE: To optimize the reconstitute process for nasopore preparation using fish scale collagen.METHODS: The optimal process for nasopore preparation through the reconstitution of fish scale collagen was ascertained by taking tilapia fish skin as a raw material to extract enzymatic soluble collagen at a temperature lower than the collagen denaturation temperature and recombinant rate of collagen fibers as index. Optimization of the conditions for nasopore preparation was carried out using single factor test and orthogonal test. The prepared nasopore was analyzed through infrared spectroscopy and microstructure analysis.RESULTS AND CONCLUSION: The optimal conditions for nasopore preparation were determined through the single factor test and orthogonal test as follows: 20 ℃ for 10 hours at pH 7.4 using a mixture of 65 mmol/L NaCl and 1 g/L collagen, by which the reconstitute rate of collagen fibers was up to 68.6%. The prepared nasopore is characterized by a refined porous structure constituted by threadlike collagen fibers, and has complete three-dimensional spiral structure,which is a potential intracavitary hemostatic material with fine properties.

Objective:Hyperoxia is a necessary therapy in some neonatal diseases, and long-term therapeutic hyperoxia may induce severe damaging effects on intestinal epithelial cells.The aim of this study was to investigate whether hyperoxia could promote the expression of ASK1 and P38 in intestinal epithelial cells through ROS.Methods:The human colon adenocarcinoma cell line Caco-2 cells were treated with different concentrations of H 2O 2(100 μmol/L, 200 μmol/L and 400 μmol/L)and 85% oxygen in vitro.The expression of ASK1 was detected by immunofluorescence, and the expression of P38 and p-P38 were detected by Western Blot and Real-time PCR. Results:With the increase of H 2O 2 concentration, the fluorescence intensity of ASK1 increased.The fluorescence intensity of ASK1 in the hyperoxia group was significantly stronger than that of the control group and the H 2O 2 groups.With the increase of H 2O 2 concentration(100 μmol/L、200 μmol/L、400 μmol/L), the expression of P38 protein(0.21±0.02, 0.28±0.13, 0.44±0.07)and p-P38 protein(0.09±0.02, 0.19±0.03, 0.37±0.07)gradually increased.The expression of P38 mRNA in 200 μmol/L and 400 μmol/L H 2O 2 groups(4.03±0.68、3.94±0.71)was significantly higher than that in 100 μmol/L H 2O 2 group(3.05±0.47)( P<0.01). The expressions of P38 protein, p-P38 protein and P38 mRNA in the hyperoxia group were significantly higher than those in the H 2O 2 group( P<0.01). Compared with the control group, the expressions of P38 protein, p-P38 protein and p38 mRNA in the hyperoxia group and H 2O 2 groups increased significantly( P<0.01). Conclusion:The expression of ASK1 and P38 in intestinal epithelial cells increased significantly under hyperoxia, which indicated that hyperoxia might activate ASK1 and thereby regulate the expression of downstream P38 through ROS, resulting in intestinal epithelial cells damage.

BACKGROUND: Studies are very few regarding the specific reaction of bone marrow mesenchymal stem cells (BMSCs) to activated microglia. Moreover, it remains unclear how MSCs maintain dopaminergic neuronal survival under specific microenvironment.OBJECTIVE: To explore the effect of BMSCs stimulated by activated microglia on dopaminergic neuron survival.METHODS: BMSCs were isolated from Wistar rats by attachment method, and in vitro cultured; microglia was activated, and dopaminergic neurons were cultured by enzyme digestion method. The experiment included 5 groups: BMSCs, microglia, lipopolysaccharide (LPS)+microglia; BMSCs+LPS+microglia groups, in which the dopaminergic neurons were cultured with corresponding culture medium; the dopaminergic neurons alone group was cultured with 10% fetal bovine serum+ DMEM/F12. The effect of different microenvironment on dopaminergic neuron survival and gliocyte-derived neurotrophic factor released from BMSCs were detected by immunofluorescence technique.RESULTS AND CONCLUSION: The release of gliocyte-derived neurotrophic factor in groups involving BMSCs was greater than corresponding control group. Tyrosine hydroxylase immunofluorescence showed that neuronal survival of dopaminergic neurons alone group was 15%, microglia group was 10%, LPS+microglia was 5%, but BMSCs+LPS+microglia group was 28%, significantly greater than the other groups (P < 0.05). In addition, survival of in vitro cultured dopaminergic neurons was decreased with increasing culture duration, but the survival of dopaminergic neurons in group involving BMSCs was significantly greater than corresponding control group. This indicates that microglia activation stimulated BMSCs to upregulate gliocyte-derived neurotrophic factor to prevent dopaminergic neurons from toxic injury, and inhibit delayed death of dopaminergic neurons.

Objective: To analyze the chemical constituents of Polygonum multiflorum extract which may cause human liver cell damage and to explore the mechanism. Methods: Raw and processed Polygonum multiflorum were extracted by 70% ethanol, then raw and processed Polygonum multiflorum water-eluted material (RW and PW), 50% ethanol-eluted material (R50 and P50) and 95% ethanol-eluted material (R95 and P95) were obtained by absorbing through AB-8 macroporous resin, followed by water, 50% ethanol and 95% ethanol elution in order. The water extracts of raw and processed Polygonum multiflorum (RWE or PWE) were obtained by boiling them in water as usual. Normal human liver L02 cells were treated by different concentrations of eluted Polygonum multiflorum materials for different time, and the cell growth inhibition of each group was determined by methylthiazolyldiphenyl-tetrazolium bromide method. The chemical constituents which had a significant cytotoxicity to L02 cells were analyzed by high-performance liquid chromatography (HPLC). Morphological changes of L02 cells were observed by Giemsa staining and cell cycle distribution was observed by flow cytometry. Results: It was found that 95% ethanol-eluted extracts of raw and processed Polygonum multiflorum showed significant growth inhibition on normal human liver L02 cells, while the other components showed no significant inhibition on cell growth. HPLC analysis showed that the main component in 95% ethanol-eluted extract of raw and processed Polygonum multiflorum was emodin at content of (18.53+/-2.96)% and (10.28+/-1.34)% respectively. Cell cycle analysis showed that 95% ethanol-eluted material of Polygonum multiflorum and emodin had a similar significant effect of S phase arrest and all could induce L02 cell apoptosis. Conclusion: The main part of Polygonum multiflorum causing liver cell damage is the 95% ethanol-eluted extract, and emodin is one of the important chemical constituents leading to liver cell damage.

BACKGROUND: The in vitro amplification is a primary method for harvesting endothelial progenitor cells (EPCs) due to its simple operation and low cost.OBJECTIVE: To isolate EPCs from rabbit bone marrow to further observe the effects of autologous EPCs on promoting vascular endothelial repair.DESIGN, TIME AND SETTING: An open experiment was performed at the laboratory of Department of Internal Medicine, Changzheng Hospital, Second Military Medical University of Chinese PLA between March 2005 and February 2006. MATERIALS: Eight New Zealand rabbits of either gender, aged 6-8 months, weighing (2.5:L-0.5) kg, were included in this study. Rabbit bone marrow was taken for isolation of bone marrow mononuclear cells by density centrifugation. METHODS: Bone marrow-derived mononuclear cells were inoculated at 1×106/cm2 and cultured for 7 days in M199 medium containing vascular endothelial growth factors and basic fibroblast growth factors. EPCs were identified by Dil-labeled acetylated low-density lipoprotein (Dil- Ac-LDL) and FITC-labeled lectin BS-1 staining. Cells that phagocytized Ac-LDL displayed red fluorescence, cells that combined with lectin BS-1 showed green fluorescence, and cells that were labeled with both exhibited orange fluorescence. Expression levels of CD133, CD134, and Flk-lwere detected using immunofluorescent staining and through the use of flow cytometer.MAIN OUTCOME MEASURES: ① Cellular morphology observation. ② Proliferative capacity of EPCs.③EPCs identified by Dil- Ac-LDL and FITC-labeled lectin BS-1. ④ lmmanohistocbemical identification of EPCs. ⑤Flow cytometry identification of EPC surface marker.RESULTS: ① Cellular morphological observation: the newly isolated bone marrow-derived mononuclear cells exhibited a round appearance. Following 72-hour culture, adherent cells grew in colony cluster, presenting with round or irregular appearance, and nuclear division was obvious. By day 7, flaky cell colonies mutually connected together, presenting with shuttle-shaped endothelioid cells.② Proliferative capability of EPCs: in the 2-4 days of culture, EPCs proliferated fast, and the proliferation slowed down thereafter, exhibiting a typical "S" -shaped appearance. By days 6 and 7, EPC proliferation accelerated again, with the absorbance values of 0.58±0.15 and 0.62±0.23, respectively. ③ Over 95% of EPC cytoplasm exhibited red fluorescence after stained with Ac-LDL, appropriately 100% of cytoplasm exhibited green fluorescence after stained with FITC-labeled lectin BS-1, and over 90% of cytoplasm exhibited orange fluorescence after double staining. ④ Immonohistochemistry and flow cytometry results revealed positive expression of EPC surface markers CD133, FIK-1, and CD34.CONCLUSION: Cell population with EPC characteristics can be successfully isolated from rabbit bone marrow by in vitro amplification.

Objective:To study the preventive and therapeutic effects of Jiechangyan Qixiao granule ( JQX) in the rats with ulcer-ative colitis ( UC) induced by dextran sulfate sodium .Methods:The UC model was induced by drinking dextran sulfate sodium ( DSS, 4%) freely in Wistar rats weighting 180-220g.Guben Yichang tablets and sulfasalazine was used as the standard drugs for the compari -son.After 7-day intragastric administration of Jiechangyan Qixiao granules at the dose of 2.70, 1.35 and 0.68 g· kg-1 · d-1 , the se-rum levels of malondialdehyde (MDA), superoxide dismutase (SOD), nitric oxide (NO), interleukin -6(IL-6)and tumor necrosis factor-α(TNF-α), and the protein expression of myeloperoxidase (MPO) and intercellular adhesion molecule (ICAM-1)in colon and nuclear factor(NF-kBp65) were detected.Results:Compared with the model group, the granule at high and medium dose could sig-nificantly increase the activity of SOD in blood and decrease the contents of MDA , TNF-αand IL-6 (P<0.05 or P<0.01).The granule could also notably decrease the MOP activity in colonic mucous of UC rats (P<0.05 or P<0.01), and the contents of NF-kBp65 and ICAM-1 in the inflammation reaction(P<0.01).Conclusion: JQX shows promising efficacy in the treatment of UC rats induced by dextran sulfate sodium .