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.

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.

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 The purpose of this study was to improve the clinical results of the treatment firearm injury to maxillofacial-cervical region, so as to raise the life quality of these patients. Methods Based on the study of the anatomical study of this region, different methods were adopted to reconstruct various maxillofacial-cervial firearms injuries. Three methods were adopted to reconstruct the mandibular region by early bone transplantation to the defects of the mandible in diffferent regions. In patients with combined defects of the mandible and soft tissue, the defects were repaired by transplantation of vascular pedicle graft with muscle and nerve or free tissue transplantation to restore function. Results The survival-rate of early bone transplantation was 91.52%(54/59). The rate of function restoration was 90.72%(88/97) in cases of tissue flap grafting. Conclusions Applied anatomy is the basic of design new surgery method. Tne new developed surgery methods can improve clinical cured effect of maxillofacial-neck region firearms injure.

Objective To study the effect of continuous renal replacement therapy(CRRT) in preventing myonephropathic metabolic syndrome(MNMS) after operation of acute arterial occlusion. Methods Twenty-four patients with acute arterial occlusion were divided randomly into 2 groups: CRRT group(n=11) and control group(n=13).The patients were treated with embolectomy or revascularization.In control group,we used conventional therapy such as anti-inflammation,expansion of blood capacity,anticoagulation,and correcting acidosis and electrolyte disorder.In CRRT group,patients were treated by continuous veno-venous hemofiltration(CVVH) with 6 h during operation and 24 h after operation. Results In control group,24 h after operation,the serum potassium,blood urea nitrogen(BUN),serum creatinine(SCr),and myoglobin(Mb) were significantly increased(P

To investigate the efficacy of the formula composed of the herbal drugs to soothe the liver, regulate qi, clarify the heart and eliminate vexation plus acupuncture for general anxiety disorder. Methods: Forty cases in the treatment group were treated with acupuncture plus herbal formula and 38 cases in the control group were treated with oral Doxepin. The assessment was conducted by Self-rating Anxiety Scale (SAS) and the Treatment Emergent Symptom Scale (TESS) before, during and after treatment. Results: The total effective rate was respectively 82.50% in the treatment group and 84.21% in the control group, with a significant difference in SAS assessment (P＜0.01) between the two groups before and after the treatment.There were no significant differences in the curative and remarkable effective rate, total effective rate and SAS assessment between the two groups (P＞0.05), but the side effect was higher in the control group than in the treatment group (P＜0.01). Conclusion: Acupuncture plus herbal formula can have a precise effective effect for extensive anxiety neurosis with mild toxic side effects.

Objective To invetigate the effect and safety of sufentanil mixed levobupivacaine on postoperative analgesia in pediatric caudal block anesthesia.Method Sixty pediatric patients (2 ～ 6 years old) who were undergoing elective abdominal surgery,such as repair hernia of high ligation,were randomly divided into three groups with 20 cases each.after intravenous induction,0.25％ levobupivacaine was injected in sacrum tube in group Ⅰ,0.5 μg/ml sufentanil mixed 0.25％ levobupivacaine and 1.0 μg/ml sufentanil mixed 0.25％ levobupivacaine were injected in sacrum tube in group Ⅱ and group Ⅲ respectively.The analgesia effect,the analgesia time,recover time and adverse reaction were observed and recorded 2,4,8,12,16,24 hours after the surgery.Results The analgesia effect in group Ⅱ、Ⅲ were significantly better than the group Ⅰ when 4、8、12 hours after the operation(P ＜0.05),and the analgesia effect in groupⅢ were significantly better than the group Ⅱ when 8 hours after the operation (P ＜ 0.05).There were no significant differences in three groups when 2、16、24 hours after the operation(P ＞0.05),the analgesia time in group Ⅱ、Ⅲ were significantly longer than the group Ⅰ (P ＜ 0.05),and the analgesia time in group Ⅲ were significantly longer than the group Ⅱ (P ＜ 0.05).There were no differences in the recovery time of three groups (P ＞ 0.05).There were no adverse reactions in three groups.Conclusions 0.5 μg/ml and 1.0 μg/ml sufentanil mixed 0.25％ levobupivacaine may be used on postoperative analgesia in pediatric caudal block anesthesia safely and analgesia effect and time were more better and longer than 0.25％ levobupivacaine singly.The analgesia effect in group with 1.0μg/ml sufentanil mixed 0.25％ levobupivacaine was the best in three groups with the fewest side effects.

Objective To investigate the effect of three kinds of low molecular weight heparin combined with methylprednisolone on the high coagulation state and renal function in patients with refractory nephrotic syndrome.Methods 140 cases of refractory nephrotic syndrome were divided into four groups, 35 cases in each group.The control group were treated with methylprednisolone.Small, medium, and large dose group in the control group on the basis of the treatment, were given low molecular weight heparin on once every other day, once daily, twice daily treatment, 3075 AxaIU per times.The improvement of renal function and high coagulation state were compared between the four groups before and after treatment.In the course of treatment, the occurrence of adverse reactions was recorded.Results Remission rate in large, medium and small dose group remission rate was higher than the control group(χ2 =4.375,P<0.05); there was no significant difference in remission rate of three different dose groups.The 24h urine protein and ALB of the four groups after treatment were significantly increased, BUN and SCr were significantly lower(P<0.05).The 24h urine protein, ALB, BUN, SCr of improvement of large, medium and small dose group were significantly better than the control group( P<0.05).The 24 h urine protein, ALB, BUN, SCr in large dose group improved significantly better than those in small dose group(P<0.05).The APTT, PT in large, medium and small dose groups after treatment increased significantly, FIB decreased significantly(P<0.05).The improvement of FIB, APTT, PT in large, medium and small dose group were significantly better than the control group after treatment ( P <0.05 ).The FIB, APTT, PT in large dose group improved significantly better than those in medium and small dose group ( P <0.05 ) .Conclusion The low molecular weight heparin combined with methylprednisolone can significantly improve hypercoagulable state and renal function in the patients with refractory nephrotic syndrome, and the large dose of low molecular weight heparin effect is the best, and no significant adverse reactions occurred.