diabeticfoot ulcer of rats induced by streptozotosin,and to find the significance of free radical scavenging and the antioxidative function.MethodsA total of 60 male wistar rats were randomized into the control group (20 rats) and diabetic model group (40 rats).The model group was induced to diabetic foot ulcer using streptozotosin tail intravenous injection,and the control group was injected equal citric acid-citric acid sodium buffer fluid instead.A rectangular full-thickness skin of dorsum of foot in the two groups was removed from model rats to construct diabetic foot ulcer.Serum zinc levels of the two groups were monitored,and immunohistochemical technique was used to detect the MT expression.ResultsSerum zinc levels in the model group( 1.17 ±0.09) was significantly lower than control group( 1.48 ±0.07),and the difference was statistically significant ( t =12.94,P ＜0.01 ).The MT expression in control group was low,while it significantly increased in diabetic model group,and the difference was statistically significant ( x2 =12.78,P ＜ 0.01 ).ConclusionsThe zinc levels and MT expression of diabetic foot ulcer rats may be associated with free radical scavenging and the antioxidative function.The increased expression of MT may play an important role in preventing the further development of diabetic foot ulcer disease.

Objective To investigate the therapeutic effect of kansui root on patients with severe acute pancreatitis(SAP). Methods Clinical data of 54 cases of severe acute pancreatitis treated with kansui root(kansui root group) were analyzed and compared with 54 cases of severe acute pancreatitis treated without kansui root (control group).Results The releivng time of abdominal pain was significantly shorter than that in control group( P

The aim of this study is to investigate the immune effects of BCG primary immunization and IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immunization on mice .We randomly divided the mice into 7 groups ,namely PBS negative controls ,BCG controls ,pcAg85A controls ,BCG primary immunization combined with Ag85A booster immunization controls ,BCG primary immunization combined with Ag85A and IL-12 booster immunization controls , BCG primary immunization combined with IL-12 booster immunization controls ,and BCG primary immunization combined with pcDNA3 .1 booster immunization controls .Implementing the immune in procedure of BCG primary immunization and cytokine IL-12 booster immunization combined with mycobacterium tuberculosis Ag85A DNA ,we observed the immune effect on mice by detecting the mice serum total IgG , specific lymphocyte proliferation and cytokine levels in 4 ,6 ,8 weeks after the last immunization .Comparing the mice immunized in the strategy of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine strengthening immunization to the mice in other groups by other immune ways ,we found that ,in BCG/Ag85A+ IL-12 groups ,IgG in-creased significantly (P＜ 0 .05) ,specific lymphocyte prolifera-ted significantly ,and after strengthening immunization IFN-γlevels ,IL-2 levels and IL-4 levels in the three periods were 128 .2 ±20.4,190.2±16.51,244.2±39.14 ;146.2±17.29,271.6±16.36and16.36±28.12 ;68.6±6.62,96.6±5.5and5.5± 10 .71 ,respectively ,which were higher than those in other groups (P＜0 .05) .It’s suggested that the immunization way of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immu-nization could significantly enhance the humoral and cellular immunity of the bodies ,and provide the basis for further study on protective effect test in animals .

AIM:To investigate the effect of inhibiting Mcl-1 gene expression on apoptosis of mouse peritoneal macrophages infected with different virulence of Mycobacterium tuberculosis using a technique of RNA interference .METH-ODS:The BALB/c mice were infected with prepared bacterium of the virulence strains of Xinjiang , H37Rv, H37Ra and BCG.Mcl-1-shRNA was applied to the mouse model of infection , and the control groups were set up .On 1 d, 3 d, 5 d and 7 d, the mouse peritoneal macrophages were collected .The expression of Mcl-1 at mRNA and protein levels was deter-mined by real-time PCR and Western blot .The apoptotic rate of peritoneal macrophages was analyzed by flow cytometry . RESULTS:The expression of Mcl-1 at mRNA and protein levels was up-regulated in the peritoneal macrophages from the mice infected with different virulence of Mycobacterium tuberculosis, and the cells from the mice infected with virulence strains of Xinjiang and H37Rv expressed higher level of Mcl-1 than the uninfected control cells (P<0.05).The expres-sion of Mcl-1 at mRNA and protein levels was reduced by RNA interference as compared with control group ( P<0.05 ) . Inhibition of Mcl-1 expression induced apoptosis of peritoneal macrophages in the mice .CONCLUSION: The Mcl-1 ex-pression at mRNA and protein levels in mouse peritoneal macrophages infected with different virulence of Mycobacterium tu-berculosis was effectively suppressed by Mcl-1-shRNA, which can induce macrophage apoptosis .

Objective:To transfect Mcl-1shRNA into Raw264.7 cells,and screen out specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene to figure out the effect of shRNA on Mcl-1 expression in murine macrophage cell line Raw 264.7.Methods: Specific shRNA was transfected into murine macrophage cell line Raw 264.7 via lipofectamine.Semi-quantitative RT-PCR and Western blot were respectively employed to test the changes in Mcl-1 mRNA level and Mcl-1 protein expressions 24 h and 48 h after the transfection ,and the silencing effects of the three pairs of specific shRNA fragments corresponding to different sites were analyzed.Results: Specific shRNA fragments at 24 h and 48 h could effectively reduce Mcl-1 mRNA and protein level ,with higher silencing effects than those of the normal group ,the lipofectamine group and the negative control group.There were statistically significant differences among them ( P<0.05 ).Among the three pairs of specific shRNA fragments corre-sponding to different sites ,Mcl-1 shRNA3 showed the most significant inhibiting effect on Mcl-1 mRNA and proteins.Conclusion:RNA interference can downregulate the level of Mcl-1 mRNA in murine macrophage cell line Raw 264.7 and greatly downregulate the expression of Mcl-1protein.Specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene have been screened out successfully.

Objective To study the expressions of myeloid cell leukemia-1 (Mcl-1 ) gene in mouse macrophages Raw264.7 and human macrophages THP-1,to screen out the cell lines with high levels of expression as the experimental cells,and based on the screening results to construct the short hairpin RNA(shRNA)eukaryotic expression plasmid targeting mice Mcl-1 gene for transfection and further screen out the shRNA expression plasmid with the most obvious effect in silencing Mcl-1 gene.Methods Semi-quantitative PCR method was used to detect the expression of Mcl-1 mRNA in the two kinds of macrophages.Western blotting was adopted to detect the expressions of Mcl-1 proteins in the two kinds of macrophages.Three different gene loci for Mcl-1 shRNA fragments were designed with small molecules interfering RNA (siRNA)software.Eukaryotic expression plasmid Mcl-1 shRNA 1-3 carrying the shRNA fragments was constructed by a company.And the eukaryotic expression plasmid vector was transfected into scavenger cells Raw264.7 mice via through the liposome.The transfection results 24 h and 48 h after transfection were observed under the inverted fluorescence microscope;Mcl-1 mRNA and protein expression were detected by real time quantitative PCR and Western blot,respectively.Results The relative expression levels of Mcl-1 mRNA and protein in mouse macrophages Raw264.7 were significantly higher than those of human macrophages (P <0.05).The shRNA expression vector constructed within 24 h and 48 h could decrease Mcl-1 mRNA and protein levels in Raw264.7 cells,especially with the most obvious silencing effect at 48 h.The 48-h transfection group differed significantly from normal group,liposome group and negative control group (P <0.05).Compared with Mcl-1shRNA1and Mcl-1shRNA2,Mcl-1shRNA3 had the strongest effect in silencing Mcl-1mRNA and protein.Conclusion We have successfully screened out experimental Raw264.7 cells and Mcl-1 shRNA eukaryotic expression plasmid which has an obvious silencing effect targeting on Mcl-1 in mice macrophages Raw264.7.

Objective To discuss the feasibility and effectiveness of transcatheter implantation of double-ring aortic valve stent through puncturing the tip of the heart under thoracotomy.Methods A novel double-ring aortic valve stent was independently designed by the authors.Three healthy goats were selected for this study.A small incision on the left anterolateral thoracic wall was made to expose the cardiac apex,than the puncturing of the left ventricular apex was performed to establish the working pathway.Guided by fluoroscopy,along a hard guide wire a double-ring aortic valve stent was inserted through a 22-French sheath to the site above the aortic valve.By utilizing the opened outer ring of the stent,the double-ring aortic valve stent was accurately placed at the bottom of the aortic valve sinus.Then,the balloon was inflated and the stent was released to substitute the original aortic valve of the experimental goat.The experiment results were evaluated immediately after the procedure.Results Transcatheter aortic valve implantation (TAVI) was successfully accomplished in all the three experimental goats.DSA was performed immediately after the procedure and anatomy evaluation indicated that the position of the implanted artificial aortic valve was satisfactory,which could replace the work of original valve.Conclusion It is technically feasible and clinically effective to use this novel double-ring aortic valve stent to perform TAVI through transapical route by puncturing the left ventricular apex.

[ ABSTRACT] AIM: To explore the effects of Mcl-1 signal pathway blockers on Mcl-1 expression, macrophage apoptosis and Mycobacterium tuberculosis in the model of mice infected with Mycobacterium tuberculosis H37Rv.METH-ODS:A mouse infection model was established by intraperitoneal injection of H37Rv suspension.The signaling pathway blockers AG490, PD98059 and LY294002 for JAK/STAT, MAPK and PI3K, respectively, were intraperitoneally injected into the mice infected with H37Rv.Cell acid-fast staining was used to observe whether the mouse peritoneal macrophages infected with H37Rv were successfully established.Immunocytochemical method was employed to detect Mcl-1 expression in the mouse peritoneal macrophages infected with H37Rv.The apoptotic rate in each group was measured by flow cytomer-ty.The scavenging capacity of apoptotic macrophages against H37Rv was determined by Mycobacterium tuberculosis colony counting.RESULTS:The result of cell acid-fast staining revealed the existence of dispersive arrangement of red short anti-acid Mycobacterium tuberculosis within infected macrophages.The result of cell immunocytochemistry showed strongly posi-tive expression of Mcl-1 protein in H37Rv infection group, AG490 treatment group and LY294002 treatment group, weakly positive expression of Mcl-1 protein in PD98059 treatment group, and negative expression of Mcl-1 protein in control group. The result of flow cytometry found that the macrophage apoptotic rate in H37Rv infection group was higher than that in con-trol group, while that in PD98059 treatment group was high than that in other groups with statistically significant differences (P<0.05).The result of Mycobacterium tuberculosis colony counting showed that PD98059 treatment had the most signifi-cant inhibitory effect on H37Rv strain.CONCLUSION: Mcl-1 signaling pathway blockers increase the apoptotic rate of macrophages infected with Mycobacterium tuberculosis H37Rv and inhibit the growth of Mycobacterium tuberculosis by inhibi-ting the signaling pathways of JAK/STAT, MAPK and PI3K, among which the MAPK has the most obvious interfering effect on Mcl-1, and leads to the highest apoptotic rate of infected macrophages and the strongest bacteriostasis.

20 years)was 8.60%(48/558) , which was more than that in the second decade of life ( 5.22% ). The incidence of painful group was 9.79%(14/143),which was more than that in foot trauma group 6.78%(40/630).The positive C sign was presented in 61.1%(33/54),posterior-type coalition in 38.9%(21/54),short talar neck sign in 61.1%(33/54)and talar beak sign in 22.2%(12/54). Conclusion The talocalcaneal coalition is a common development abnormality in our country. We must pay attention to the diagnosis of talocalcaneal coalition for painful foot adulthood.

Objective To study the incidence and radiologic findings of calcaneonavicular coalition.Methods CR films of foot andankle in 1361 cases were presented,which were evaluated for acute trauma or chronic pain.There were 588 cases of foot CR and 773 cases of ankle CR,age ranged from 10 years to 91 years(984 cases of 20~40 years).The prevalence of calcaneonavicular coalition was determined and the different significance of both male and female,acute trauma and chronic pain group were analysed.Results In 1361 cases,72 cases of calcaneonavicular coalition(5.3%) were demonstrated,8.7%(47/588)on foot CR films and 3.2%(25/773) on ankle CR films.Calcaneonavicular coalition was more dipicted on foot CR films than on ankle CR films(P0.05).Conclusion The foot CR films is more superexcellent than the ankle CR films on demonstrating calcaneonavicular coalition.