Objective To construct baculovirus expression system for the human plasminogen kringle 5(hK5) gene,and detect its expression in Spodoptera frugiperda 21(sf21). Methods Baculovirus Bacmid-rhK5 was prepared with bac-to-bac baculovirus expression system,and sf21 cells were infected.The expression of rhK5 in sf21 cells was determined by Western blot.The activity of purified protein was detected by vascular endothelial cell inhibition test. Results The pFastBac HTB-rhK5 was successfully constructed,and sf21 cells were transfected with the constructed vector.The output of rhK5 obtained was 90 ug/L,and the protein possessed the inhibitory activity of endothelial cell proliferation.The median effective dose(ED50) was 4 ug/mL. Conclusion The rhK5 baculovirus expression vector is highly expressed in sf21 cells,which lays a foundation for the the expression of rhK5 protein with biologic activities.

Objective: To elucidate the effect of cervical multi-infarction on memory function. Method: Homogenous microthrombi were injected into internal carotid artery(ICA) through external carotid artery of Sprangue-Dawlay rat to get unilateral,bilateral or recurrent embolic cerebral multi-infarcction model. Water maze and passive-avoidance tests were used to assess the memorial function. Brain histopathological changes were also conducted. Result: Memorial function was severely injured,on the contrast to the pseudo-operational rats. Among the three models, unilateral,bilateral and recurrent ones, recurrent multi-infarction model is the worst on the memorial function, while the bilateral group is worse. Conclusion: Cerebral multi-infarction can cause memorial dysfunction. The degree of memorial function is closely related to the degree of ischemic severity.

Objective To investigate the protective effect of adiponectin on angiotesin Ⅱ (Ang Ⅱ)-mediated cardiomyocyte apoptosis,and explore the related mechanism. Methods Cardiomyocytes isolated from neonatal rats were cultured in vitro. The identity of cardiomyocytes was confirmed by morphological examination and staining with anti-sarcomeric ?-actin antibody,and most (more than 95%) of the cells were identified as neonatal rat ventricular myocytes. After being placed in a serum-free medium for 24 hours,the ventricular myocytes were randomly grouped and received the following treatments respectively:placebo,or Ang Ⅱ (1?mol/L),or Ang Ⅱ (1?mmol/L) plus adiponectin (30mg/L). The protein levels of Bcl-2 and Bax were detected by Western blotting. The apoptotic rate and the levels of intracellular reactive oxygen species (ROS) in the ventricular myocytes were determined by flow cytometry. The survival rate of ventricular myocytes was evaluated by trypan blue staining. Results As compared with placebo group,the apoptosis of neonatal rat ventricular myocytes was significantly promoted in 1?mol/L angiotesin Ⅱ group,the survival rate of cardiomyocytes was significantly reduced,with the protein expression of Bax and the ROS level increased,and the protein expression of Bcl-2 decreased (P

Objective To explore clinical significance of subtemporal decompression in traumatic total anterior circulation infarcts(TTACI).Methods 24 TTACI cases accepted subtemporal decompression were analyzed including CT scan feature, pre operative observation to temporal superficial artery, operative cranial pressure classification and blood circulation in cortical small artery.Results Clinical symptom compared between before and after operation: good 4 cases, unchanged 14 cases, dead 6 cases. CT scan showed infarct areas became smaller in 4 cases, unchanged in 9 cases, larger in 2 cases three months after operation.Conclusion Subtemporal decompression cannot reduce the areas of infarcts and mortality, nor improve neurological function and promote quality of survivor.

In the light of requirements of medical equipment simulation system, the overall framework of joint control system of medical equipment integrated support in the theater is designed in this paper. Such modules are developed with Delphi as communication module, module for equipment basic information, module for equipment video information and module for medical faculty information. A joint control system suiting integrated support simulation sand table for medical equipment is complteted,which provides a multidimensional informational platform for integrated demonstration of medical equipment.

To investigate the inhibitory effects of siRNA expression vector on proto-oncogene Pokemon expression in SW480 cells and to provide experimental basis for further research about the biological function of Pokemon.siRNA expression vectors were constructed to express a short hairpin RNA against Pokemon.The recombinants were transfected into SW480 cells with liposome.Cellular morphology and transfection efficiency were observed.The expression of Pokemon were checked by real-time fluorescence quantitative PCR and Western blot.MTT assay was used to detect the effect of siRNA on the growth of SW480 cells and Fluorescence Activated Cell Sorter was applied to study cell apoptosis of SW480 cells.The transfection efficiency was about 36% and the cellular morphology changed greatly at 48h after transfection.siRNA expression vectors could specifically reduce the expression of Pokemon mRNA and protein in SW480 cells.Compared with negative control,the inhibition ratio of Pokemon mRNA expression was 34.2% and 67.7%,in 24th hour and 48th hour,the inhibition ratio of Pokemon protein was 48.3% and 73.6%,in 48th and 72th hour,respectively.Transfection with Pokemon siRNA expression vectors could inhibit cell growth and induce cell apoptosis of SW480 cells.siRNA expression vectors were successfully established that could effectively inhibit the expression of Pokemon in SW480 cells.Pokemon gene silencing could decrease growth and increase apoptosis of SW480 cells.

0.05).Conclusion:So-fastTM can greatly improve the penetration of PS-ODN into S.mutans and can be used as appropriate delivery system of PS-ODN for S.mutans. No matter what approaches were adopted, the uptake rate reached the maximum at 5 ?mol/L and 2 h-exposure. The penetration can not be enhanced by increasing the PS-ODN concentration and the transformation time.

Background Protein kinase B (Akt) is the center of multiple cellular signaling pathways,and it participates in the regulation of cell function,such as proliferation,migration,and metabolism of cells.Phosphoinositide 3-kinase (PI3K) promotes activation of Akt and therefore triggers many signal pathways.PI3K inhibitor can silent Akt,but whether it can affect the biological behavior of lens epithelial cells (LECs) during the posterior capsular opacity (PCO) is worthy of investigation.Objective This study was to explore the effect of LY294002,a PI3K inhibitor,on Akt activation in human LECs.Methods Human LECs strain,HLEC-B3 cells,were cultured and passaged.The cells were incubated to 96-well plate for 24 hours,then LY294002 was added with the final concentration at 0,10,20,30,40,50,60,70,80 μmol/L,respectively.After incubation for 24 hours,cell counting kit-8 (CCK-8) was used to detect the inhibitory rate of cell proliferation.Transforming growth factor-β2 (TGF-β2) of 10 μg/L was added in the medium of cells as TGF-β2 group,TGF-β2 + LY294002 (20 μmol/L) was used to coculture the other group of cells,and the cells without TGF-β2 and LY294002 served as the control group.Phosphorylation of Akt (p-Akt) in the cells was detected by laser scan confocal immunofluorescence microscope.The expression level of p-Akt was evaluated using Western blot.Results CCK-8 assay showed that the A value of the cells was gradually reduced with the increase of LY294002 concentration (Fgroup =9.72,P =0.00),but the A value was significant raised along with the lapse of time (Ftime =1737.54,P=0.00).Confocal immunofluorescence revealed that a little of p-Akt was expressed in the control cells.After induced by TGF-β2,lots of red fluorescence of p-Akt was seen in cells around the cell membranes.But in the TGF-β2+LY294002 co-culture cells,the fluorescence of Akt was much weaker.Western blot showed that the expression level of p-Akt was 0.91±0.08,1.48±0.13 and 0.95±0.19 in the control group,TGF-β2 group and TGF-β2 +LY294002 group,respectively,with a significant difference among the three groups (F =15.04,P =0.00).Conclusions LY294002 can inhibit the activation of Akt induced by TGF-β2.LY294002 may have utility in the prevention and treatment of PCO.

Objective To investigate the value of high frequency ultrasound and CT in the diagnosis of laryngeal carcinoma. Methods Sixty-nine cases of laryngeal carcinoma were typed and staged by high frequency ultrasound and CT, and compared with the results of laryngoscopy and postsurgical pathology. Results The typing accuracy was ~94.2 % for ultrasonography and ~91.3 % for CT, higher than that of laryngoscopy(~73.9 %).T staging accuracy of ultrasonography was T_1 ~85.9 %,T_2 ~88.5 %,T_3 ~94.1 % and T_4 ~88.9 %, higher than that of CT (~68.8 %) and laryngoscopy (~75.0 %) in T_3 stage and superior to laryngoscopy (~66.7 %) in T_4 stage. Lymph node metastases were found in 37 cases (~53.6 %) by ultrasonography, higher than those by CT scanning (~27.5 %). Color Doppler blood flow laryngeal carcinoma was shown in 65 cases(~94.2 %),most were low resistance spectrum pattern. Conclusions High frequency ultrasonography is superior to CT and laryngoscopy in the staging of laryngeal carcinoma and is of great value in choosing the therapeutical protocol.

ObjectiveTo investigate the effect of heart-regulating and mind-calming acupuncture on the expression of gap junction protein connexin43 (Cx43) in myocardial cells in a rabbit model of ventricular arrhythmia.MethodA rabbit model of ventricular arrhythmia was made by injecting barium chloride into the marginal ear vein. Forty New Zealand rabbits were randomized into control, model, acupuncture and medication groups. The acupuncture group received heart-regulating and mind-calming acupuncture[at Lingtai(GV10), Shendao(GV11), bilateral Neiguan(PC6)and Baihui(GV20)]and the medication group, oral gavage of propafenone. Rabbit left ventricle was taken after the completion of the experiment. Cx43 expressions were compared and analyzed usingan immunohistochemical staining technique and a computer image processing system.ResultCx43 expression decreased significantly in the model group compared with the control group (P<0.01) and increased in the acupuncture and medication groups compared with the model group (P<0.05). There was no significant difference in Cx43 expression between the acupuncture and medication groups (P>0.05).Conclusion Adjusting Cx43 expression in myocardial cells may be one of the target points for this acupuncture treatment of arrhythmia.