Objective:To investigate the effect of NF-κB p65siRNA of IL-17 inducing the expression of MCP-1 in the primary cultured cardiac myocytes.Methods:The cardiac myocytes were isolated from neonatal mice by different adhesion method.NF-κB P65 siRNA was transfected into cardiac myocytes and the rates of transcription and translation of MCP-1 were detected by RT-PCR and enzyme-linked immunosorbent assay( ELISA) ,the rates of transcription and translation of P-P65 and P65 were detected by RT-PCR and Western blot.Results:Compared with negative siRNA group,the expression of the MCP-1 at mRNA and protein levels were increased in negative siRNA +IL-17 group(P<0.05).Compared with negative siRNA group,the expression of the MCP-1 at mRNA and protein levels were decreased in NF-κB P65 siRNA group( P<0.05).Compared with negative siRNA+IL-17 group,the expression of the MCP-1 at mRNA and protein levels were decreased in NF-κB P65 siRNA+IL-17 group(P<0.05).The expression of the NF-κB P65 at mRNA and protein levels in cardiac myocytes were specifically and extensively suppressed by NF-κB P65 siRNA(P<0.05).After stimulated by IL-17,the amount of P-P65 in the cardiac myocytes was significantly increased in time-dependent manner compared with that of black group( P<0.05) , but the levels of P65 changed little ( P>0.05 ) .Conclusion: IL-17 stimulates MCP-1 expression in cardiac myocytes via NF-κB activation,and NF-κB P65 siRNA can effectively inhibit the upregulation of IL-17 on MCP-1.

Objective To investigate the effect of dichloroacetate on the expression of Kv1.5 in a rat model of pulmonary arterial hypertension (PAH) .Methods Thirty-two male SD rats weighing 200-250 g were randomly divided into 4 groups ( n = 8 each): normal control group (group C), dichloroacetate control group (group D),PAH group, and PAH + dichloroacetate group (group PD). PAH was induced by left lung resection combined with subcutaneous injection of monocrotaline 60 mg/kg in PAH and PD groups. In group PD, dichloroacetate 80 mg/kg was given through a gastric tube into stomach once a day for 28 consecutive days after monocrotaline injection,while the equal volume of normal saline was given instead of dichloroacetate in group PAH. Group D only received dichloroacetate 80 mg/kg through a gastric tube into stomach once a day for 28 consecutive days. Pulmonary arterial pressure (PAP) was measured at day 28 after monocrotaline injection. The rats were then sacrificed and lung tissues were removed to calculate the percentage of thickness of the tunica media of pulmonary artery and right venicular hypertrophy index and to determine the proliferating cell nuclear antigen (PCNA) and Kv1.5 protein expression (by Western blot) and Kv1.5 mRNA expression (by RT-PCR).Results Compared with group C, the PAP,percentage of thickness of the tunica media, right ventricular hypertrophy index were significantly increased, Kv1.5 mRNA and protein expression was down-regulated and PCNA expression was up-regulated in groups PAH and PD ( P ＜ 0.05). Compared with group PAH, the PAP, percentage of thickness of the tunica media, right ventricular hypertrophy index were significantly decreased, Kv1.5 mRNA and protein expression was up-regulated and PCNA expression was down-regulated in group PD (P ＜ 0.05). There was no significant difference in the indexes mentioned above between group C and group D ( P ＞ 0.05). Conclusion Dichloroacetat alleviates PAH through upregulating Kv1.5 expression in lung tissues and inhibiting pulmonary vascular remodeling in rats.

BACKGROUND: Mesenchymal stem cells (MSCs) can differentiate into osteoblasts in the action of dexamethasone; Meanwhile, bone morphogenetic protein-2 (BMP-2) can promote the cell proliferation and differentiation, and matrix secretion in the process of bone repair. BMP-2 plays an important role in treating fracture and bone defects by inducing bone formation both in vivo and in vitro.OBJECTIVE: To analyze whether dexamethasone induction in vitro could enhance the ability of BMP-2 modified MSCs in osteogenic conversion.DESIGN: A randomized paired design.SETTING: Xijing Hospital of the Fourth Military Medical University of Chinese PLA.MATERIALS: The experiments were carried out in the Orthopaedic Oncology Institute of Chinese PLA, the Fourth Military Medical University of Chinese PLA from February to August in 2004. Twenty 20-month-old New Zealand rabbits were provided by the experimental animal center of the Fourth Military Medical University of Chinese PLA [Certificate number: 2005C00117]. The rabbits were raised normally at room temperature with normal humidity. The samples were collected bilaterally, the cells from the left limb were taken as the dexamethasone-induced group, and those from the right limb as the control group.METHODS: MSCs treated and untreated with dexamethasone were transfected with BMP-2 gene, then the BMP-2 expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical staining. The growths of the MSCs were observed, and the activity of alkaline phosphatase (ALP) and osteocalcin of MSCs were determined with ALP kit and osteocalcin radioimmunoassay kit after Ad-BMP-2 transfection.MAIN OUTCOME MEASURES: ① BMP-2 expression in MSCs; ② Morphological changes of the MSCs; ③ ALP activity and osteocalcin content in the MSCs. RESULTS: ① Expressions of BMP-2 gene could be observed in MSCs both treated and untreated with dexamethasone after transfection. ② The forms were irregular in most of the MSCs treated with dexamethasone, appeared as triangular or polygonal changes.They grew more slowly than those cultured with basic medium. There were no obvious changes of cellular forms after gene transfection. ③ Five days after transgene, the ALP activity in the MSCs supernatant in the dexamethasone-induced group was higher than that in the control group [(134.36±8.84), (104.02±7.83) nkat/L, t =3.350 6, P ＜ 0.01, n =20]. The amount of osteocalcin secretion in MSCs in the dexamethasone-induced group was higher than that in the control group [(14.68±0.73), (6.52±1.21) μg/L, t =3.568 2, P ＜ 0.01, n =20].CONCLUSION: Dexamethasone induction before transgene can promote the proliferation of BMP-2 modified MSCs and the conversion into osteoblasts.

Objective To investigate and compare the killing effects of different prodrugs combined with suicide gene HSV-tk on laryngocacinoma cell, Hep-2 in vitro. Methods Retroviral expressing vector pL(tk)SN was constructed by recombinant DNA technology. Hep-2 cells were infected by the recombinant retrovirus. The positive cloning was obtained after G418 selection and were termed Hep/tk. The integration and expression of tk gene in Hep-2 cells were identified by RT-PCR and Southern blot. The growth state and prodrugs killing effect of tk gene modified cells were used to investigate the expression of tk gene and antitumour effect on Hep-2 cells. Results RT-PCR and Southern blot analysis confirmed the integration and expression of tk gene in Hep-2 cells. There was no significant difference in cell proliferation between the Hep/tk and Hep-2. After the treatment of GCV,the Hep/tk showed high sensitivity to GCV and bystander effects were observed siginificantly in vitro. However the efficiency of another two prodrugs ACV and BVdU was lower than that of GCV. Both tk-positive and tk-negative Hep-2 cells were relatively insensitive to ACV and BVdU.Treatment of tk gene modified cells mixed with different proportion parental cells shoewd obviously bystander effects.Conclusions The laryngocarcinoma cells Hep-2 have sensitive to HSV-tk/GCV system and have significant bystander effects, which might have therapeutic potential value for laryngocarcinoma.

Biocontrol bacteria 308R(pKSH) with hrpN of Erwinia amyloyora. It can secrete the Harpin protein which can induce plant resistance. After successive growth in antibiotic-free LB medium for 50 generations, only 23.1% of the cell still retained the plasmid pKSH, in comparison with 4.75% of the cosmid pCPP430. Sprayed onto the leaves of tomato, the recombinant strain maintained a population density of over 10~ 5 cfu/cm~ 2 when kept under high humidity in thirteen days, but if without humidity, the strain keep 10~ 4 cfu/cm~ 2 over in five days. On the tomato leaves, the stability of the recombinant strain 308R(pKSH) was higher than the strain 308R(pCPP430). The experiment indicated that the recombinant strain 308R(pKSH) had higher stability compared with the strain 308R(pCPP430), but that was not enough. In this paper reasons for unstability and strategies for improving were discussed for the recombinant strain.

Objective To study the clinical and electrophysiological characteristics of peripheral neuropathy in patients with multiple sclerosis(MS).Methods As a total of 84 MS patients,the clinical and electrophysiological data were analyzed in 16 MS patients with the symptoms of peripheral neuropathy(PNMS group)and in 68 MS patients without peripheral neuropathy(NPNMS group).Results The incidence of peripheral neuropathy was 19.0%(16/84)in this group.The clinical symptoms included extremity numbness in 14 cases(87.5%),limb inertia in 12 cases(75.0%),and radicular pain in 4 cases(25.0%).Signs included decrease of tendon reflex in 15 cases(93.7%),periphery sensory disability in 13 cases(81.2%),and myatrophy in 5 cases(31.2%).The mean age in the PNMS group was(44.6?12.5)years vs.(32.2?11.5)years in the NPNMS group.The course of disease was(39.3?18.3)months vs.(31.6?17.2)months in the NPNMS group,there were significant differences in two groups(P

Bone marrow transplantation was considered effective treatment for leukemias and other haematological diseases. Acute graft v-host disease (GVHD) is a major complication of allogeneic bone marrow transplantation. The T lymphocytes were specially depleted by anti-CD5, CD2, CD8, CD27 monoclonal antibody-ricin immunotoxins. Inhibition of the protein synthesis and T-cell functions in the target cells was observed in vitro. At 10 9mol / L of the immunotoxin, 3 logs (99.9%) of target cells were killed, but no cytotoxicity on nontarget cells. At the same concentration, the anti T cell immunotoxin had no any influence on the proliferating rate of CFU-GM and BFU E. None of the ten patients who received T cell depleted bone marrow developed grade III or IV acute GVHD.

Objective: To investigate which step of IL-6 signal transduction pathways in myeloma cells can be taken as the acting target of the antagonists for IL-6. Methods: EMSA and immunoprecipitation were used to detect the activation of transcription factors(TFs)-STAT3, NF-IL-6 and protein kinase ERK in a myeloma cell line-Sko-007 by IL-6. then anti-sense expression plasmids and anti-sense ODN for these signal moleculars were constructed and designed,the effects of these anti-sense nucleic acids on IL-6 signal transduction in Sko-007 cells were analyzed by the same methods. Results: IL-6 signal transduction could be antagonized by these anti-sense nucleic acids at different extent.Conclusion: TFs or protein kinases in IL-6 signal trareduction pathways can be taken as the target for antagonists design.

Objective To investigate the clinical role of a combination therapy of percutaneous radiofrequency ablation(RFA) and transcatheter arterial chemoembolization(TACE) in large hepatic tumors. Methods Out of 62 patients with hepatocellular carcinomas confirmed by pathology, 21 patients received a combination therapy of TACE and RFA(combination group), 22 patients TACE therapy alone, and the rest 19 patients RFA therapy alone. A total of 106 tumors with a mean diameter of ( 5.9? 0.7) cm(ranged from 5.0 to 8.1 cm) were detected, and the largest tumor was selected for observation in a patient with multiple lesions. There was no significant difference in mean age, tumor size and liver function grade among the three groups.Results Tumor complete necrosis accounted for 80.9%in combination group, which was significantly higher than that of TACE group and RFA group ( 27.2%, 47.4%,P 0.05). Mean survival duration of combination group was 25.6 months, significantly higher than that of TACE group( 14.9 months)(P0.05). Conclusions Compared with TACE or RFA therapy alone, the combination therapy improves tumor complete necrosis rate and prolongs the patients′ survival duration.

Objective　To investigate the IL-6 signal transduction pathways and their regulation mechanism in a human myeloma cell line-U266. Methods　Electrophoretic mobility shift assay (EMSA) was used to detect the activation of the transcription factors (TFs)-STAT3 and NF-IL-6 by IL-6. Cells were treated with chemical agents or transfected with the expression plasmids for the two TFs or the anti-sense oligonucleotide for protein kinase involved in one IL-6 signal transduction pathway. The change of the activation state of another IL-6 signal transduction pathway was also exhibited by EMSA. Results　Two IL-6 signal transduction pathways (JAK/STAT and Ras/NF-IL-6) can be activated by IL-6 of different dose in U266 cells. When one of the two signal transduction pathways was up-regulated, the other one was down-regulated. Conclusion　There is an antagonistic effect between the activation of two IL-6 signal transduction pathways in U266 cells.