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Table of Content

    05 September 2013, Volume 33 Issue 9
    Establishment and identification of a Tet-on HeLa cell model of CX26/CX32 gap junction
    2013, 33(9):  1079-1084. 
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    Objective To establish a specific cell model for fast and efficiently measuring the effect of candidate drug on gap junction (GJ), which will provide an ideal experimental tool for both the drug discovery targeting GJ and the specific research for GJ. Methods The bidirectional vector pBI plasmid under the control of a bidirectional doxycycline-inducible promoter was constructed. The Tet-on HeLa cells were transfected with CX26/CX32 cDNA and the cell model stably expressing CX26/CX32 and functional heteromeric GJ channels were established. Cells were cultured in the presence and absence of doxycycline (Dox, 1 μg/mL), the expression of CX26 mRNA and protein in Tet-on HeLa cell were assayed by RT-PCR and Western blot, respectively. The GJ function between adjacent HeLa cells was detected by dye transfer assay, and the proliferation of HeLa cells was measured using sulforhodamine B (SRB) assay. Upon the above cell model, the effects of GJ inhibitor, 2-aminoethoxydiphenyl borate (2-APB), and activator, retinoid acid (RA), on the function of GJ were also observed. Results HeLa cells expressed no endogenous CX, and expression of CX26 mRNA and protein in Tet-on HeLa cells could be induced by the addition of Dox in the culture medium. Formation of functional GJ due to CX induction by Dox was observed as evidenced by the dye transfer assay. 2-APB (50 μmol/L) decreased dye spread through GJs composed of CX26/CX32 in HeLa cells with a GJ inhibition rate of 51.3% (P<0.01); while RA (10 μmol/L) increased dye spread through GJs between adjacent HeLa cells with a GJ enhancement rate of 60.3% (P<0.01). Conclusion Tet-on HeLa cell model stably expressing CX26/CX32 induced by Dox was successfully established, providing an useful technology for both the drug discovery targeting GJ and the specific research of GJ modulation by candidate drugs.
    Down-regulation of GINS2 Inhibits Cell Cycle Regulators in HL60 Cells
    2013, 33(9):  1085-1090. 
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    Objective To observe changes of cell cycle regulators after down-regulation of GINS2 in HL60 cells lines and investigate related mechanism. Methods The recombinant plasmid carrying siRNA and non-homologous sequence were stably transfected into HL60 cells as experimental group and negative control group respectively, while, blank control group only treated by lipidosome and HL60 cells acted as untreated group. Flow cytometry was used to detect the cell cycle of HL60,3H-TdR incorporation assay and Colony-forming test were performed to measure duplication and nucleic acid synthesis in four groups either. The protein expression of cycle regulatory proteins CDK1 and CyclinB1 were measured by Western blot. Meanwhile,transcription and translation levels of ATM,CHK2,P53 were detected by RT-PCR and Western blot respectively. Results 48h after transfection, GFP could be observed by Fluorescence microscope. Flow cytometry showed that cell growth was significantly inhibited and G2 peak was found in interfering group. Compared with other three groups,nucleic acid synthesis and cell proliferation was blocked in experimental group. CDK1,cyclinB1 related with cell cycle regulation were dramatically decreased over time. Together,both RT-PCR and Western blot demonstrated that the transcription and translation levels of ATM,CHK2,P53 were notably increased. Conclusions Down-regulation of GINS2 can suppress nucleic acid duplication and influence cell cycle process of HL60 by up-regulation of ATM,CHK2 and P53.
    Adenovirus-mediated overexpression of BMP9 inhibits proliferation and migration in human osteosarcoma cell line 143B
    2013, 33(9):  1091-1096. 
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    Objective The aim of this study is to investiage the effect of bone morphogenetic protein 9(BMP9) on the biological behavior of human osteosarcoma cell line 143B.Methods Using RT-PCR and Western blot annlysis to characterize the endogenous expression of BMP9 in the osteosarcoma cell line. Using a recombinant adenovirus expressing BMP9(adBMP9)to infect the osteosarcoma cell line with relatively low endogenous BMP9,then Transwell assay was used to determine cell invasion; Wound-healing assay was used to determine cell migration;MTT assay and crystal violet staining were used to determine the influence of adBMP9 on osteosarcoma cell proliferarion; Hoechst 33258 assay was used to determine cell apoptosis;Clonoy formation assay was used to determine cell clonoy. Results Osteosarcoma cell proliferation、migration、invasion and colony formation were significantly decreased while the cell apoptosis was induced by adBMP9. Conclusion adBMP9 is able not only to inhibit cell proliferation and migration but also induce cell apoptosis in human osteosarcoma cell line 143B.
    Up-Expression and of SPARC in db/db mice kidney
    2013, 33(9):  1097-1100. 
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    Objective To test db/db mouse kidney tissue of the cysteine-rich acidic secreted proteins(SPARC)expression. Methods RT-PCR, Western blot and immunofluorescence method detection db/db mouse kidney tissue of SPARC mRNA and protein expression. Results SPARC highly expressed in db/db mouse kidney tissue. Conclusion SPARC in the db/db mouse kidney tissue showed high expression may be related to the occurrence and development.
    Activation of TLR3 inhibits angiogenic factors expression in the human trophoblast cells
    2013, 33(9):  1101-1106. 
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    Objective: To investigate the effect of TLR3 ligation on expression of angiogenic factors in primary trophoblast cells and evaluate the possible role of TLR3 signal pathway in the pathogenesis of hypertensive diorders in pregnancy. Methods The first trimester human trophoblast cells and immortalized human trophoblast cell line Swan71 were stimulated with Poly(I:C) (TLR3 specific ligand). sFlt-1 and PlGF protein and mRNA expression were evaluated by ELISA and Real-time PCR respectively at different time points. TLR3 mRNA expression was also determined by Real-time PCR. Results 24,48 and 120 h after Poly(I:C) stimulation, concentration of sFlt-1 in the conditioned media of Swan71 was significantly increased compared with non-treatment control (P < 0.05). Poly(I:C) (10 μg/mL) significantly induced sFlt-1 mRNA (P < 0.05), while suppressed PlGF mRNA (P < 0.05) in primary trophoblast cells. Induced expression of sFlt-1 mRNA showed a time and dose-dependent manner, which reached the highest effect 2 h after Poly (I:C) stimulation (P < 0.05). At the same time, PlGF mRNA expression was decreased significantly (P < 0.05). Expression of TLR3 mRNA was significantly increased 8 h up to 12 h after Poly(I:C) stimulation (P < 0.05). Conclusion TLR3 activation induced synthesis and secretion of sFlt-1, while inhibit PlGF in trophoblast cells, which lead to disturbance of angiogenesis and be involved in the pathophysiology of hypertensive disorders in pregnancy.
    Promotion of the invasion and proliferation of trophoblast cells by silencing Fzr1
    2013, 33(9):  1107-1111. 
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    Objective To explore the spatial and temporal expression of Fzr1 protein during the development of placenta, and observe the effect of Fzr1 on the invasion and proliferation of trophoblast cells. Methods To observe the expression level and localization of Fzr1 protein using Western blot and immunohistochemical methods, and detect the effect of Fzr1 on trophoblast cell invasion and proliferation by RNA interference technology, Transwell and MTS methods. Results (1) Immunostaining revealed that Fzr1 protein was strongly expressed in the cytotrophoblast and syncytiotrophoblast cells of villi during early pregnancy. Positive signals were mainly located in the cytoplasm. In the third trimester of pregnancy, Fzr1 protein was expressed in the syncytiotrophoblast, and expression levels significantly decreased. Positive signal was located in the nucleus. Western blot studies showed that Fzr1 protein levels in the third trimester decreased by 4.88 times compared with early pregnancy (P<0.01). (2) Transwell studies revealed that Fzr1 siRNA significantly enhanced the invasion of JAR cells compared with blank control (P<0.01), but scrambled siRNA had no significantly change. (3) MTS results showed that Fzr1 siRNA significantly enhanced the proliferation of JAR cells compared with blank control (P<0.01), but scrambled siRNA had no significantly change. Conclusions Fzr1 protein was temporally and spatially expressed at the placenta of normal pregnancy, and Fzr1 gene silencing could promote the invasion and proliferation of trophoblast cell.
    Genetic stability and proliferation capacity of human bone marrow mesenchymal stem cells in pulmonary adenocarcinoma microenvironment
    2013, 33(9):  1112-1117. 
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    Objective To investigate the genetic stability of human bone marrow mesenchymal stem cells (HMSC-bm) under pulmonary adenocarcinoma microenvironment. Methods A co-cultured system of HMSC-bm and A549 cells was established through the combination of 6 well culture plate and transwell chamber as experimental group while HMSC-bm and A549 cells were cultured separately as the control groups. The morphology of the cells was observed by phase-contrast microscopy. The proliferation curve of mesenchymal stem cells was tested by MTT. Chromosome was examined by karyotyping analysis. The expression of HDAC4 protein was detected by Western blot. Results Cells in the co-culture group showed that the cell nucleus became bigger than the nucleus in HMSC-bm group and exhibited anachromasis in 7 days. The proliferation curve indicated faster proliferation of co-cultured HMSC-bm than HMSC-bm. Karyotyping analysis showed that the chromosome number of co-cultured HMSC-bm was hypotriploid and triploid and the chromosome was grossly abnormal. The expression of HDAC4 protein in co-cultured HMSC-bm significantly increased compared to that in HMSC-bm (p<0.01). Conclusion Pulmonary adenocarcinoma microenvironment could influence the proliferation speed and the genetic stability of HMSC-bm .
    Effect of embryoid body adhesion time on the cardiac differentiation of mouse embryonic stem cells
    2013, 33(9):  1118-1122. 
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    Objective To investigate the time-dependent effect of embryoid body (EB) adhesion on the cardiac differentiation of mouse embryonic stem cells (ESCs) and its mechanisms. Methods Hanging drop culture was used to induce ESCs differentiating into cardiomyocytes through forming EBs. Suspended EBs were transferred onto tissue culture plates at different differentiation days. The percentage of beating EBs was calculated at different time points. The mRNA expression of Nkx2.5, GATA4 and β-MHC were detected by RT-PCR. The phosphorylation of Src kinase was detected by Western-blot. Results The groups whose EB adhesion times were at differentiation day 3 or 4 had higher percentage of beating EBs and higher gene expression of Nkx2.5, GATA4 and β-MHC compared with those groups whose EB adhesion times were at differentiation day 5, 6 or 7(P<0.05). EB adhesion triggered the phosphorylation of Src kinase, which was inhibited by PP2(P<0.05). Furthermore, the percentage of beating EBs and gene expression of β-MHC were enhanced by PP2 at differentiation day 4~6 in the group whose EB adhesion time was at differentiation day 4(P<0.05). Conclusion Our results indicate that the EB adhesion time is important in regulation of the cardiac differentiation of ESCs. When EBs adhere to gelatin-coated plates at differentiation day 4 or earlier, the cardiac differentiation is significantly inhibited. The mechanism is likely that EB adhesion triggers the phosphorylation of Src kinase.
    TanⅡA triggered the differentiation of NB4 and MR2 cells by up-regulated expression of C/EBPβ
    2013, 33(9):  1123-1128. 
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    Objective:To Investgiate the expression of C/EBPβ in APL cells during differentiation induced by TanⅡA. Methods TanⅡA treated NB4 and MR2 cells 120h, the characterizations of differentiation were examined by morphology and membrane differentiation antigens;The mRNA and protein expressions of C/EBPβ in different concentration TanⅡA-intervened APL cells were examined by RT-PCR and Western-blot;APL cells were treated with different concentration TanⅡA, the characterizations of differentiation were examined by membrane differentiation antigens. Results TanIIA induced the differentiation of ATRA-sensitive and ATRA-resistant APL cells(p<0.05); TanIIA dose-dependend upregulated the expression of C/EBPβin mRNA and protein(p<0.05); TanIIA promoted the differentiation of the NB4 and MR2 cells in a dose-independent manner and exerted its strongest effect at a concentration of 1 mg/L. Conclusion TanIIA promoted the differentiation of ATRA-sensitive and ATRA-resistant APL cells and exerted its strongest effect at a concentration of 1mg/L; C/EBPβ played a key role in TanIIA-induced differentiation.
    In vivo and in vitro killing effect of the TSA on ovarian cancell cells
    2013, 33(9):  1129-1134. 
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    Objectives To investigate the killing effect and mechanism of histone deacetylase inhibitor, trichostatin A (TSA), on ovarian cancer. Methods The killing effect of TSA on ovarian cancers was detected by using XTT and flow cytometry technique on two of the normal ovarian epithelial cell lines, IOSE-29 and IOSE-329, and three of the ovarian cancer cell lines, ES-2, SKOV-3, and OVCAR-8. The in vivo effect was also observed. Results HDAC6 protein showed higher positivity in ovarian cancer and its cell lines than in normal ovarian epithelia. ES-2, SKOV-3, and OVCAR-8 ovarian cancer cells were more sensitive for HDACs inhibitor, TSA, treatment compared with normal ovarian epithelial cell lines (P<0.05). Also, the ratio of apoptosis and cell death were significantly higher in TSA treated ovarian cancer cells than it in the untreated group. In vivo experiments also showed that ES-2 xenograft treated by TSA was growing much slower compared with untreated ES-2 xenograft. Conclusions TSA could effectively kill the ovarian cancer cells in vivo and in vitro, and the mechanism might be carried out partially by HDAC6 gene inhibition.
    Expression of WAVEs in ovarian cancer and its implication
    2013, 33(9):  1135-1140. 
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    Objective To investigate the expression of WAVEs in ovarian cancer(OC)and clinical pathological characteristics. Methods The expressions of WAVEs were detected by immunohistochemistry technique in 60 cases of OC,30 ovarian benign tumors,25 ovarian borderline tumors and 21 normal ovarian tissues.The expression of WAVEs was subjected to statistical analysis combined with patients’ clinical pathological data. The protein and mRNA levels of WAVEs were detected by real time PCR (RT-PCR) and western blotting respectively. Results 1)The expression of WAVE1 in OC was significantly higher than ovarian normal tissues, ovarian benign tumors and ovarian borderline tumors, P<0.01. 2)The expression of WAVE1 in OC was closely related with clinical stage、pathological differentiation and Ca-125 level (P<0.05) .Whereas there was no relevance to age,tumor size or volume of ascites(P>0.05). 3)The expressions of WAVE1 mRNA and protein in OC were significantly higher than ovarian normal tumor,ovarian benign tumor and ovarian borderline tumor,(P<0.01;Both WAVE2 and WAVE3 have very low expression of mRNA and protein level in all groups. Conclusion Overexpression of WAVE1 may play an essential role in the development of OC and may be bound up with the invasion and metastasis of OC. That may be a potential new therapeutic target for OC.
    bFGF-2 inhibits bone morphogenetic protein9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells
    2013, 33(9):  1141-1145. 
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    Objective Observe the effect of FGF2 in BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells,and investigate its mechanism.Methods C3H10T1/2 cells were infected by adenovirus recombined with FGF2 and BMP9,then the early osteogenic maker alkaline phosphatase(ALP) activity were detected by quantitative and staining assay,later osteogenic maker calcium deposition was determined by Alizarin Red S staining,p12SBE-luc activity were detected by quantitative,Smad1/5/8,Runx2 and OCN were determined by western blot. Results BMP9-induced early osteogenic marker ALP, and late osteogenic markers matrix mineralization and osteocalcin (OCN) are inhibited by FGF2, BMP-Smad1/5/8 pathway and Runx2 are inhibited by FGF2.Conclusion FGF2 inhibits bone morphogenetic protein9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells.
    Changes in autophagy and its role in rat lung following Ischemia/reperfusion injury
    2013, 33(9):  1146-1149. 
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    Objective To explore the level and the role of autophagy in lung ischemia-reperfusion injury. Method Animals were subjected sham operation or ischemia-reperfusion. We detected the LC-3 protein in lung tissue by western blot and observed autophagosomes by electronic microscope. To explored the role of autophagy in lung I/R injury, 3-methyladenine (3-MA) were used to pretreat the Animals. HE stain and blood analysis were used to evaluated lung injury. Result The results indicated that the autophagic flux was elevated during ischemia period, and was significantly enhanced during reperfusion. Inhibition of autophagy by 3-methyladenine (3-MA) ameliorated lung I/R injury, as indicated by index number of lung injury and blood analysis. Conclusion The results demonstrated autophagy might be a scathing factor in lung I/R injury.
    Heme oxygenase - 1 / carbon monoxide system inhibits angiotensin II induced apoptosis of myocardial cells
    2013, 33(9):  1150-1154. 
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    Objective To investigate the influence of heme oxygenase -1/carbon monoxide (HO-1/CO) system on apoptotic myocardial cells induced by angiotensin II (AngII) . Methods Primary culture of myocardial cells in neonatal Wistar rat, randomly divided cells into four groups. Respectively with Real Time - PCR and Western blot analyzed HO -1 mRNA and protein expression, colorimetric method was used for determination of cell culture supernatant fluid medium carbon oxygen hemoglobin content; flow cytometry instrument was used to detect cells apoptosis rate. Results AngII group of myocardial cell HO -1 mRNA, protein, COHb content and cell apoptosis were significantly higher than those in the control group (P < 0.05), AngII+hemin group HO -1 mRNA, protein and COHb content further raise (P < 0.05), while apoptosis rollback (P < 0.05), but still higher than those of the control group (P < 0.05), and AngII+ZnPPIX group only apoptosis significantly increased (P < 0.05), other index no significant change. Conclusion HO-1/CO system attenuate the myocardial cells apoptosis induced by AngII.
    Wnt/β-catenin signaling pathway in inhibition of Raw264.7 cell differentiation
    2013, 33(9):  1155-1159. 
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    Objective To learn the regulation of Wnt/β-catenin signaling pathway in the mouse monocyte/macrophage Raw264.7 differentiation and maturation. Methods The cells were divided into four groups: the negative control group (empty Raw264.7), the experimental control group (infected with Ad-GFP), the positive control group (50ng/mL RANKL induced) and the experimental group (infected with Ad-β-catenin). RNA were extracted after the cells were treated for 3, 6, 9days respectively, The expressions of nuclear factor KB receptor, Tartrate-resistant acid phosphatase, Cathepsin K and Metalloproteinase-9 gene were detected by fluorescence quantitative RT-PCR respectively. Cells treated for 6 days were used to observe the differentiation of Raw264.7 by Tartaric acid-resistant phosphatase (TRAP) staining and expressions of TRAP and Cathepsin K protein by Western blot. Results qRT-PCR showed that Ad-β-catenin group could decrease the mRNA expression of RANK, TRAP, Cathepsin K and MMP-9 gene. TRAP staining showed that 50ng/mL RANKL could successfully induced Raw264.7 into multi-cores. Few multinucleated cells in the control group and the Ad-GFP group were seen, In Ad-β-catenin group cells were mononuclear. The TRAP and Cathepsin K protein expression was significantly reduced analysed by Western blot ( p<0.05). Conclusion Wnt/β-catenin signaling pathway can inhibit Raw264.7 differentiate into mature osteoclasts.
    The relationship between Ccbe1 expression and VEGF-C, lymphangiogenesis as well as prognosis in human laryngeal carcinoma
    2013, 33(9):  1160-1164. 
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    Objective To investigate the expression of Ccbe1 in patients with laryngeal cancer, and the relationship with vascular endothelial growth factor C, lymphangiogenesis and their prognosis significance. Methods Forty five specimens of the laryngeal cancer and twenty benign laryngeal diseases were studied.The protein expression of Ccbe1 and VEGF-C were observed by immunohistochemistry and Western bolt. LVD was evaluated by LYVE-1 staining, and then judged the role of Ccbe1 in laryngeal cancer prognosis by using Kaplan-meier method. Results The protein expression of Ccbe1 and VEGF-C were much higher in tumor tissue than in the normal tissue(P<0.05), and the expression of Ccbe1 was significantly correlated with VEGF-C(r=0.338,P<0.05).The expression of Ccbe1 was significantly correlated with LVD, TNM stage and lymph node metastasis(P<0.05),and the expression of VEGF-D was significantly correlated with LVD and lymph node metastasis(P<0.05).The different expression of Ccbe1 had effect on survival ratio(P<0.05).Conclusion Ccbe1 stimulates VEGF-C upregulation and promote lymphangiogenesis in laryngeal cancer. Detection of Ccbe1 protein may be helpful for judging prognosis in laryngeal cancer.
    Effect of CD45RO N-glycosylation sites on galectin-3 binding to CD45RO mutant-transfected cells
    2013, 33(9):  1165-1170. 
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    Objective To construct eleven cell lines each of which expressed CD45RO with removal of one N-glycosylation site and to examine its binding to galectin-3. Methods Individual site-directed N→Q mutagenesis of N-glycosylation site was performed for all eleven N-glycosylation sites in CD45RO and cloned into lentiviral vector pWPXL. Recombinant lentiviral was produced by 293T cells with co-transfection of pWPXL-mutated-CD45RO, the packaging plasmids psPAX2 and MD2.G. Human J45.01 cells were infected by the recombinant lentiviruses. The expression of CD45RO mutants was confirmed by RT-PCR and flow cytometry, and the binding of these cell lines to galectin-3 was evaluated by flow cytometry. Results Eleven cell lines each of which expressed CD45RO with removal of one N-glycosylation site were successfully constructed. RT-PCR, DNA sequencing and flow cytometry analysis revealed that the mutations were correct and CD45RO with removal of one N-glycosylation site was stably expressed on the surface of each cell lines. It was observed that galectin-3-binding to N327Q mutant was largely enhanced, to N36Q or N217Q mutant was largely suppressed. Conclusion Galectin-3 binding to CD45RO-J45.01 cells is regulated by N-glycosylation sites in CD45RO.
    Exenatide improves cardiovascular function in diabetic rats
    2013, 33(9):  1171-1175. 
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    Objective To investigate the effect of Exenatide (Ex), a glucagon-like peptide-1 (GLP-1) receptor, on cardiovascular function in diabetic rats, and to provide the theoretical foundation for the clinical therapy. Methods A completely randomized design with four treatments was employed in this study. Thirty four Wistar rats were randomly divided into four groups, including control (C, n=7); diabetic model (DM, n=9,); low dose of Ex intake (Emin, n=9);. high dose of Ex intake (Emax, n=9). The level of fasting plasma glucose was determined. Ejection fraction (EF) and the fractional shortening (FS) of the left ventriculus of the rats were determined by using ultrasound for small animals. The blood velocity of abdominal aortic flow (AF) was measured. The injuries in the endangium of thoracic aorta were inspected by using the thoracic scanning electron microscope. Results The level of fasting plasma glucose was significantly higher (P<0.01) in rats from DM group than from control group. The rats in exenatide treatment group had a lower level of fasting plasma glucose than those in DM group (P <0.05). The rats of DM group had a more striking dysfunction either in ejection fraction or fractional shortening compared with control group (P <0.01), and attenuated levels than exenatide treatment group (P <0.05). In addition, the blood velocity of abdominal aortic flow in rats from DM group was obviously slower in control group (P <0.01), and vascular intimals of rats from former group were tremendously damaged. The blood velocity of abdominal aortic flow in rats from exenatide treatment group was much faster than DM group, the level of vascular intimal injury was alleviated. Conclusion In addition to reducing the level of glucose, exenatide can also ameliorate cardiovascular dysfunction in diabetic rats.
    Photodynamic therapy under endoscope on patients with early and earlier esophageal carcinomas
    2013, 33(9):  1176-1179. 
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    Objective To evaluate effectiveness of photodynamic therapy(PDT) under endoscope in 10 patients with esophageal carcinomas, investigate the preventions of its side-effectiveness. Methods 10 cases with esophageal carcinoma were included in this study, which accepted PDT during June, 2011 and June, 2013. Forty-four to forty-eight hours after intravenous injection of photosensitizer (photosan) (2mg/kg), semiconductor laser irradiation was performed by gastric endoscope to local tumor tissues. Treated for two consecutive days as a cycle, and repeated another cycle one month later. 1,3,6,12,18,and 24 months later after PDT, gastroscope, chest and abdominal enhanced CT-scan were taken to evaluate the effectiveness and side-effectiveness of PDT, and the managements of these adverse effects were investigated. Results Tumor tissues occurred edema and necrosis on the following day after PDT, part or all of which disappeared one month later. One month later after the second cycle of PDT, tumor tissues in all of the 10 cases disappeared under gastric endoscope, and tumor cell invisible under microscope. From a follow-up of 2-24 months in the 10 cases, no recurrence or metastasis was observed under gastric endoscope, chest and abdominal enhanced CT-scan and serum tumor markers. Except for photosensitivity reactions, the main adverse effects of PDT were transient fever, chest pain, cough and expectoration, pulmonary infection and dysphagia, all of which remissed through symptomatic treatments. 2 cases presented esophageal stenosis, and then remitted through endoscopic dilation and stent placement. Hypercoagulable state in vary degrees were observed in all of these 10 cases, more over, one of them got acute coronary syndrome. Conclusions For early staged esophageal carcinoma, the effectiveness of PDT was definite. The adverse effects of PDT were relatively moderate and could be remitted through symptomatic treatments. The reason and mechanism of abnormal coagulation function still need further investigation.
    Hemeoxygenase-1 up-regulates of MMP-2 and MMP-9 expressions in liver of spontaneously hypertensive rats
    2013, 33(9):  1180-1185. 
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    Objective To explore the effects of hemeoxygenase-1 (HO-1) induction on matrix metalloproteinase 2 (MMP-2) and MMP-9 expression in liver of spontaneously hypertensive rats (SHR). Methods Rats were randomly divided into 4 groups. Wistar induction group (WH) and SHR induction group (SH): 8 g/L hemin was administered to the animals intraperitoneally at a concentration of 15 mg/kg/d for 4 consecutive days to induce expression of HO-1; Wistar control group (W) and SHR control group (S): equal volume of 0.1 mol/L NaOH (pH 8.3) was administered intraperitoneally as controls. Systolic blood pressure (BP) of rats was determined. The expressions of HO-1, MMP-2 and MMP-9 in liver were determined by immunohistochemistry. The relative activity of MMP-2 and MMP-9 in plasma were detected by gelatin zymography. Results The expressions of liver HO-1, MMP-2 and MMP-9 were significantly higher in WH and SH group compared with W and S group, respectively (p<0.01). Induction of HO-1 significantly attenuated BP of SHR. The relative activity of MMP-2 and MMP-9 in rat plasma was significantly higher in S group compared with W group (p<0.01). The relative activity of MMP-2 and MMP-9 in rat plasma was significantly higher in WH and SH group compared with W and S group, respectively (p<0.01). Conclusions HO-1 up-regulates expressions of MMP-2 and MMP-9 in liver of SHR and the mechanisms need to be further elucidated.
    Transplantation of placenta-derived mesenchymal stem cells reduces hypoxic-ischemic brain injury in rats
    2013, 33(9):  1186-1190. 
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    Objective In the present study, we investigate the neuroprotective effect of placenta-derived mesenchymal stem cells (PDMSCs) transplantation in rat's hypoxic-ischemic brain damage(HIBD) model and its mechanism. Methods The rats were randomly divided into four groups: sham-operated group; only HIBD group; HIBD+PDMSCs-treated group and HIBD+Fibroblasts-treated group. After 2 weeks of treatment, we measured inflammatory cytokines (TNF-a, IFN-γ, IL-10 and IL-17) expression in the brain and serum by appropriate methods. Results Our results indicated that pro-inflammatory cytokine gene and protein expressions increased in the HIBD group, and that PDMSCs-treated group significantly (all P<0.05) lowered at these levels. Furthermore, PDMSCs treatment was able to increase IL-10 levels. We also observed that motor dysfunctions in Fibroblasts-treated group were not improved and the indicators were not statistically significant compared with PDMSCs-treated group. Conclusions In summary, PDMSCs treatment plays a critical role in the neuroprotection observed in this model and the mechanism may be partly through alleviating inflammatory reaction.
    Effect of FK506 on NIT-1 cells proliferation and insulin secretion
    2013, 33(9):  1191-1194. 
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    Objective To investigate the effects of FK506 on cell proliferation of pancreatic NIT-1 beta-cell and insulin secretion function. Methods Effect of FK506 on proliferation and apoptotic were mensured by MTT-assay, Flowcytometry; Insulin secretion was investigated by Radioimmunoassay and Real-time PCR. Results FK506 (20μg?L-1) could inhibit the proliferation of NIT-1 cells, and could induce apoptosis; FK506(10μg?L-1) could inhibit NIT-1 cell insulin release, The mRNA expression of PDX-1 and GLUT2 were down-regulated. Conclusion FK506 inhibit NIT-1 cells insulin secretion, the mechanism maybe related to the down-regulation of. PDX-1 and GLUT2.
    Continuous infusion of rocuronium with close-loop auto-feedback system in hepatic surgery
    2013, 33(9):  1195-1198. 
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    Objective To assess the clinical effect of continuous infusion of rocuronium with close-loop auto-feedback system on laparotomic hepatic surgery. Methods 30 patients (ASA I/II) who underwent hepatic surgery were randomized divided into continuous infusion of rocuronium group (group A, n=15) and intermittent bolus infusion group (group B, n=15). Recovery index, antagonistic time, recovery time and the average amount of rocuronium were compared between the two groups under muscle relaxants monitoring. Surgeons’ satisfactory scores were recorded. Results There was no significant difference inrecovery index between the two groups (P = 0.59> 0.05), while antagonistic and recovery time of group A wereshorter than those of group B (P <0.01), and both correlated with rocuronium average amount (at 0.01 level).Patients in both groups did not appear any respiratory complications or any other adverse events after extubation. Group B had higher satisfactory scores from surgeons (P <0.01). Conclusion Continuous infusion of rocuronium with close-loop auto-feedback system in hepatic surgery was safe. It reduced rocuroium’s amount, and could reduce the risks of residual neuromuscular blockade.
    Growth factor receptor binding protein-2 inhibits colon cancer HT29 cell proliferation
    2013, 33(9):  1199-1204. 
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    Objective To analyze the effect of Grb2 inhibition by shRNA transfection on the proliferation of colon cancer cells HT29,and to estimate the feasibility of Grb2 as a target for the treatment of colorectal cancer. Method The Grb2 shRNA was transfected to 293T cells by shRNA lentiviral vector system, 5 strains of different sequence Grb2 shRNA lentiviral particles was obtained and infected colon cancer HT29 cells respectively. 5 colon cancer HT29 cell lines infected with the Grb2 shRNA lentiviral particles for the experimental group (i.e. infected the Grb2 shRNA cells HT29/shGrb2-69,HT29/shGrb2-70,HT29/shGrb2-71, HT29/shGrb2-72,HT29/shGr-b2-73),eGFP shRNA slow virus infection colon cancer HT29 cell for the negative control group. MTT method was used to determine the cell proliferation situation, the expression of Grb2 mRNA was detected by RT-PCR and Grb2、P42/44 ERK、phosphorylation P42/44 ERK、serine/threonine protein (Akt)、phosphorylated Akt (P-Akt)、STAT5 and other molecules signaling pathway detected by Western Blot. Results 72h after infection,HT29/shGrb2-69, HT29/shGrb2-73 cell were suppression in the mRNA and protein levels. After 24h infection ,HT29/shGrb2-69、HT29/shGrb2-73 cell A values were 0.176±0.045、0.186±0.013,and 48h for 0.347±0.048,0.382±0.041 (Significantly lower than the blank control, negative control, P<0.001),and 72 hours after infection were 0.934±0.038、0.983±0.205 (compare with the blank control, negative control, P<0.01); 72h after infection,Phospho-P42/44 ERK, Akt, Phospho-Akt were decreased, while the ERK,STAT5 expression were not affected. Conclusion The application of a lentiviral vector system transfected of Grb2 shRNA to HT29 cells inhibited the levels of Grb2 mRNA and protein significantly, and induced HT29 cell proliferation and down-regulated signal transduction molecules expression.
    Short-term effects of Infliximab and Cyclosporin as rescue therapy for patients with refractory-IBD
    2013, 33(9):  1205-1208. 
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    Objectives To evaluate the effect of Infliximab(IFX) and cyclosporin(CsA) as rescue therapy for patients with refractory-IBD. Methods Prospectively and not randomly, 18 cases with refractory-IBD were invited to participate in the study. Of 12 patients with UC, 6 cases received IFX; 6 patients took CsA. All of 6 patients with CD were given IFX. Comparing the effect of IFX with CsA at 12 weeks and 30 weeks. Results 5 UC cases responded to IFX and CsA, 1 case failed to therapy respectively for both groups at 14 weeks. Of 6 UC cases with IFX 3 were effects and 1 failed , but 4 cases responded and 1 case failed for CsA group at 30 weeks. Of 6 CD cases, 3 responded to IFX while 1 case failed at 14 weeks. Of 6 CD cases treated with IFX, 4 patients improved, no invalid case was found at 30 weeks. Rash occurred in 2 cases treated with IFX, while among cases treated with CsA, 1 developed tatter and 4 had hands tremor. Conclusions As rescue therapy for patients with refractory IBD, IFX and CsA presented the same effectiveness, Patients with good economic conditions had better take the fomer, those with relatively poor economic bear ability might as well choose the latter.
    Antimicrobial peptide P18 inhabits the proliferation of Hep-2 cells
    2013, 33(9):  1209-1211. 
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    Application of molecular imaging in stem cell therapy for ischemic heart diseases
    2013, 33(9):  1214-1218. 
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    Stem cell therapy might ameliorate heart failure by promoting neovascularization, regenerating cardiac tissue, and recruiting resident stem cells. Several clinical trials have shown beneficial effects of stem cell transplantation to improve cardiac function in ischemia heart disease. However, to fully understand the beneficial effects of stem cell therapy, investigators must be able to track the biology and physiology of transplanted cells in living subjects over time. Molecular imaging, a multidisciplinary field which provides integrated information on specific molecules of interest within the cells of living subjects and thus holds great promise as an effective way to track the transplanted cells.
    A preliminary exploration: how to build the Academic Health System in China
    2013, 33(9):  1219-1222. 
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    With the development of the society, the changing functions of academic medicine, result in institutional expansion and integration in the health field. The institutional net is called academic health system, which integrate various all kinds and levels of health institutions centering the universities、medical schools and its affiliated hospitals, and have more functions of education、research and medical care. It will be a hot focus in the field of health.