Objective:To construct a small interfering RNA(siRNA)expression vector(psiRNA-VEGFR-3)targeting vascular endothelial growth factor receptor 3(VEGFR-3)and to investigate the effects of VEGFR-3 siRNA on the adherence and invasion of human colon cancer cells.Methods:A siRNA expression vector(psiRNA-VEGFR-3)targeting VEGFR-3 were constructed and was used to transfect LoVo cells via lipofectamine 2000.The mRNA and protein expression of VEGFR-3 were examined after transfection by reverse transcriptase polymerase chain reaction(RT-PCR)and Western blotting,respectively.The tumor adhesion ability was detected by cell-matrix adhesion experiment and the invasion ability of tumor cells was evaluated by millicell chamber model.Results:The VEGFR-3 siRNA expression vector was successfully constructed.The expression of VEGFR-3 mRNA and protein was inhibited after psiRNA-VEGFR-3 transfection.Seventy-two hours after psiRNA-VEGFR-3 transfection,Western blotting assay showed that the expression of VEGFR-3 protein was decreased from(1.26?0.19)to(0.39 s0.12)(P

Objective:To evaluate the effects of Er,Cr:YSGG laser combined with NaF treatment on the acid resistance of human dental hard tissue.Methods:The enamel or dentin samples were randomly divided into four groups and prepared by different methods as follows: ①without treatment(control group),②laser irradiation(Laser group),③ treated with 20 g/L NaF solution(NaF group),④laser irradiation+ NaF solution treatment(Laser and NaF group).The samples of each group were respectively demineralized in 0.1 mol/L of lactic acid for 24 h and the calcium ion in the demineralization solution in each group was measured by atomic absorption spectrophotometry.The fluoride on the surface of the samples was examined by SED-X and the ultrastructure was investigated by SEM.Results:The concentrations of Ca~(2+) dissolved from the enamel or dentin samples in NaF,laser and NaF and laser groups were significantly lower than that in the control group(P(0.05)).Only in the NaF group,the F peak appeared.SEM observation showed the lased surface showed various patterns of microirregulation with a scaly appearance,the openings of dentinal tubules were visible,but no melting or carbonization;there were lot of particles attached to the surface of NaF treated samples;the ultrastructure of the NaF and laser-treated sample surfaces was similar to that of the laser treated.Conclusion:The combined use of laser and NaF dose not have synergetic effects on acid resistance of human dental hard tissue.

Objective:To observe the influence of schisandrone on the levels of IL-1? and iNOS mRNA in the hippocampus of rats with Alzheimer-like disease(AD) and to investigate the probable prevention and therapy effect of schisandrone on AD. Methods: Totally 30 male SD rats, 8-12 weeks old, weighing 250-300 g, were randomly divided into 3 groups: blank control group,AD model group and schisandrone intervention group(n=10). The animal model of AD was established by 10 mmol/L amyloid-beta protein(A? 25-35 ) 5 ?l stereotactic injection into lateral cerebral ventricle of rats and the rats of schisandrone intervention group were administrated with corn oil with 1 mmol/L schisandrone. And then the levels of IL-1? and iNOS mRNA in the hippocampus were detected by RT-PCR. Results: The levels of IL-1? and iNOS mRNA in the hippocampus of rats with Alzheimer-like disease (0.333 7?0.122 3, 0.266 6?0.088 5) decreased obviously after administration of schisandrone (0.969 3?0.153 9, 0.666 0?0.121 1)(P

Objectives This study was designed to observe the correlation of the levels of serum N-terminal pro-brain natriuretic peptide(NT-proBNP) and high sensitivity C-reactive protein(hs-CRP) between the 3rd day after acute myocardial infarction(AMI) and after 3 months of left ventricular remodeling(LV remodeling) and to establish the predictive levels of NT-proBNP,hs-CRP for LV remodeling after AMI.Methods The blood samples from 106 patients with the first AMI and echocardiography were examined on the 3rd day and after 3 month.LV remodeling was estimated by the changes of left ventricular end-diastolic volume(LVEDV) during the 3rd day and after 3 month.Based on the change of LV remodeling,the 106 patients were divided into two groups:LV remodeling group(defined as the change rate of LVEDV more than 20%) and non-LV remodeling group.Results The correlation coefficients of the change of LVEDV were 0.706 for serum NT-pro BNP and 0.596 for hs-CRP.With a cutoff value of 0.20,the area under the ROC curve(AUCs) was(0.892) for NT-proBNP and 0.825 for hs-CRP.There was no statistically difference between NT-proBNP and hs-CRP.The ROC plot indicated that NT-proBNP was superior to hs-CRP for predicting LV remodeling.Conclusion The level of serum NT-proBNP appropriately reflects LV remodeling after AMI and shows more efficacious than the level of serum hs-CRP.The level of serum NT-proBNP on the 3rd day after AMI and LV remodeling were positively correlated with the level after 3 months.

Objective:To discuss the relationship between the serum level of interleukin-6 (IL-6) and the presence of an intrauterine contraceptive device (IUCD). Methods:The experimentalgroup was subdivided into the bleeding and the non-bleeding groups which consisted of 64 womenwith IUCD. There were 60 healthy women in the control group. The serum levels of IL-6 and C-reactive protein (CRP) were detected by enzyme-linked immunosorbent assay (ELISA) in bothgroups. Results :In the experimental group,the serum levels of IL-6 in 3 cases were increased,andthere was a significant difference in comparison with those of the control group (P＜0. 01),butthere were no statistical differences between the bleeding group and the non-bleeding group withIUCD (P＞0． 05). Serum CRP was negative both in the experimental groups and in the controlgroup. Conclusion:The non-bacterial endometrial inflammation induced by IUCD causes the in-crease of serum level of IL-6 in some women. It is suggested that IL-6 may play an important rolein the contraceptive mechanism of IUCD.

Objective To evaluate the effects of dexmedetomidine on the expression of aquaporins 1 (AQP1) and AQP5 during lung ischemia/reperfusion (I/R) in rats in an in vitro experiment.Methods SPF healthy male Sprague-Dawley rats,aged 3-4 months,weighing 250-320 g,were used in this study.Forty-five isolated rat lungs in which the model of isolated lung perfusion was successfully established were divided into 3 groups (n=15 each) using a random number table:sham operation group (S group),I/R group and dexmedetomidine group (D group).The isolated rat lungs were continuously perfused for 150 min in group S.After 15 min of perfusion,the isolated rat lungs were subjected to 60 min of ischemia and apnea followed by 75 min of ventilation and reperfusion to establish the model of isolated lung I/R injury in group I/R.In group D,the isolated rat lungs were perfused for 15 min with K-H perfusion fluid containing 10 nmol/L dexmedetomidine,and then subjected to 60 min of ischemia and apnea followed by 75 min of ventilation and reperfusion with K-H perfusion fluid containing 10 nmol/L dexmedetotnidine.The lung compliance,airway resistance,perfusion flow and partial pressure of arterial oxygen (PaO2) were recorded at 10 min of perfusion and 15,45 and 75 min after restoration of perfusion.Lung tissues were obtained at 75 min after restoration of perfusion for determination of wet/dry weight ratio (W/D ratio) and for examination of the pathological changes and changes in the uhrastructure.The expression of AQP1 and AQP5 protein and mRNA in lung tissues was detected by Western blot and real-time polymerase chain reaction,respectively.Results Compared with S group,the airway resistance was significantly increased and lung compliance,perfusion flow and PaO2 were decreased during reperfusion,and W/D ratio was increased in I/R and D groups (P＜0.05),the expression of AQP1 and AQP5 protein and mRNA in lung tissues was significantly down-regulated in group I/R,and the expression of AQP 1 and AQP5 protein and mRNA in lung tissues was significantly up-regulated in group D (P＜0.05).Compared with I/R group,the airway resistance was significantly decreased and lung compliance,perfusion flow and PaO2 were increased during reperfusion,W/D ratio was decreased,the expression of AQP1 and AQP5 protein and mRNA in lung tissues was up-regulated (P＜0.05),and the pathological changes of lung tissues was significantly attenuated in group D.Conclusion The mechanism by which dexmedetomidine reduces I/R injury may be related to up-regulation of the expression of AQP1 and AQP5 in rats in an in vitro experiment.

Objective To explore the engrafted effect of NSCs combined with OECs on the motor function repair of hindlimbs in rats subjected to spinal cord transected injury.Methods The NSCs and OECs were isolated from the Transgenic Green Fluorescent Protein(GFP) embryo mouse' hippocamp and neonatal mouse' olfactory bulb.Rats were divided into NSCs group,OECs group and co-transplantation group as well as control group.The recovery of posterior limb motor function in rats subjected to spinal cord transected injurywas evaluated by the BBB locomotion score respectively in each weekend among eight weeks.This was followed by a histochemiscal observation to know the engrafted cells survival and migration in the spinal cord of host under the fluorescence microscope.Results The NSCs and OECs cells survived and migrated in the spinal cord of transplantation groups:the motor functional improvement of hindlimb in rats was seen in all cell transplanted groups;NSCs group,OECs group and co-transplantation group demonstrated a significant increase than that of control group at 3 and 4 weeks post operation(P 0.05).Conclusions The engrafted NSCs,OECs and co-transplantation can improve partly the motor function recovery of hindlimb in rats at 3 to 4 weeks following cord transection;The OECs group and co-transplantation have better effects than that of NSCs group,but there is no obvious difference between the OECs group and co-transplantation group at observed time point in this investigation.

AIM: To investigate the effect of Zhongfeng Nao De Ping, a traditional chinese medicine, on the cerebral ischemia/reperfusion injury in rats. METHODS: Delayed neuronal death (DND) model was established by bilateral carotid arteries occlusion for l0 min followed by 1 day and 7 days reperfusion. Effects of sodium glutamate, ketamine and Zhongfeng Nao De Ping on DND in the hippocampus was examined by counting neuronal density in CAl Sector, and the contents of Ca 2+ and amino acids in the hippocampus were also measured. RESULTS: The contents of the glutamate and aspartate in the hippocampus were increased significantly at l0 min after transient cerebral ischemia. Sodium glutamate markedly decreased the neuronal density in hippocampal CAl sector, while ketamine and Zhongfeng Nao De Ping increased it significantly in the same sector. The level of Ca 2+ increased on day 1, day 7 after the reperfusion, while Zhongfeng Nao De Ping inhibited the increase in Ca 2+ concentration in hippocampus. CONCLUSION: These results suggested that Zhongfeng Nao De Ping has an effective protection against DND in hippocampus following cerebral ischemia/reperfusion in rats.

Objective:This study aimed to observe the synergistic effect of a new tumor vaccine combined with metronomic che-motherapy in vivo on breast cancer. This study was also conducted to investigate the mechanism of this combination. Methods:Balb/c mice inoculated with 4T1 mouse breast cancer cell were used as tumor models. High-mobility group nucleosome-binding protein 1 (HMGN1) gene was used to transfect 4T1 cell lines as cancer vaccines. After 4T1 cell was inoculated, the mice were randomized into four groups:normal saline (NS);metronomic gemcitabine (GEM) alone;cancer vaccine alone;and combination therapy group. Tumor growth and potential toxicities of these regimens were observed. The Foxp3 expression of regulatory T cells (Tregs) was detected by western blot and immunohistochemical staining. The microvessel density (MVD) of the tumor was also detected by immunohistochemi-cal staining. Results:The tumor volume of the mice was significantly lower in the combination group than in the MET group or cancer vaccine group (P<0.05). This result exhibited a higher significant difference than the tumor volume of the mice in the NS group (P<0.01). Foxp3 expression was significantly lower in the mice treated with GEM (combination or MET group). MVD was significantly lower in these two groups than in the cancer vaccine group or NS group (P<0.05). Furthermore, adverse reactions slightly occurred in each group. Conclusion: The combination of cancer vaccines and metronomic GEM is a very active and well-tolerated regimen for breast cancer in mice.

Objective To investigate the expression of Sema4 D and Ki67 in pancreatic carcinoma and its relationship with clinicopathological features. Methods The expression of Sema4 D and Ki67 in 40 pancreatic carcinoma tissues and 10 adjacent tissues was measured by immunohistochemistry, and its relationship with the clinicopathological features of pancreatic carcinoma was analyzed. Comparison of categorical data between two groups was made by chi-square test. Spearman's rank correlation analysis was used to identify the correlation between these variables. Results Sema4 D and Ki67 were detected in 19 (47. 5％ ) and 18 (45. 0％ ), respectively, of the 40 pancreatic carcinoma tissues, with significantly higher positive rates than the adjacent tissues (χ2= 10. 040 and 10. 000, P = 0. 020 and 0. 022) . The abnormal expression of Sema4 D and Ki67 differed significantly between patients with different tumor stages and between patients with and without lymph node metastasis (χ2= 6. 352, 4. 912, 12. 031, and 6. 465, P = 0. 013, 0. 028, 0. 001, and 0. 025) . Sema4 D expression was positively correlated with Ki67 expression (r = 0. 347, P = 0. 028) . Conclusion The expression of Sema4 D and Ki67 is elevated in pancreatic carcinoma compared with the adjacent tissue. Their expression is positively correlated with each other and is associated with tumor staging and lymph node metastasis.