Objective To evaluate the antitumor effect of dacarbazine (DTIC) on B16-F1 melanoma after CDK2 gene silencing.Methods Cultured B16-F1 melanoma cells were divided into 4 groups:control group receiving no treatment,CDK2-shRNA group infected with a recombinant lentivirus pUL-CDK2-shRNA,DTIC group cultured in 96-well plates followed 12 hours later by the treatment with 250 μmol/L DTIC,CDK2-shRNA + DTIC group infected with pUL-CDK2-shRNA followed 12 hours later by the treatment with 250 μmol/L DTIC.MTT assay was performed to evaluate the growth inhibition of B16-F1 melanoma cells,and coefficient of drug interaction (CDI) was calculated.AnnexinV-FITC/PI double staining was conducted to detect cell apoptosis.C57BL/6 mice were subcutaneously injected with B16-F1 cells at exponential growth phase into the right groin to establish melanoma-bearing mouse models.Twenty mouse models were randomly and equally divided into 4 groups:control mouse group injected with phosphate-buffered solution (PBS) into tumors,CDK2-shRNA mouse group injected with pUL-CDK2-shRNA into tumors,DTIC mouse group injected with DTIC into the abdominal cavity,and CDK2-shRNA + DTIC mouse group treated with pUL-CDK2-shRNA and DTIC.The animal experiment lasted 18 days,and the tumor growth curve was drawn.After 18-day treatment,all the mice were sacrificed,and tumors were isolated and weighed.The tumor growth inhibition rate was calculated,and the tumor cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL).Results After 72-hour culture,compared with the control group,the CDK2-shRNA group,DTIC group,and CDK2-shRNA + DTIC group showed significantly decreased relative cell survival rates (40.6％ ± 2.8％,45.2％ ± 3.7％,28.7％ ± 2.1％,respectively;F =458.04,P ＜ 0.05),but significantly increased cell apoptosis rates (25.1％ ± 3.3％,15.6％ ± 2.2％,45.6％ ± 3.5％,respectively;F =115.46,P ＜ 0.05).Additionally,CDK2-shRNA + DTIC group showed significantly lower relative cell survival rates (P ＜ 0.01),but higher cell apoptosis rates (P ＜ 0.01) compared with the DTIC group.The CDI value was less than 0.7.On the sixth day after the in vivo treatment,the tumor volumes in the control mouse group,CDK2-shRNA mouse group,DTIC mouse group,and CDK2-shRNA + DTIC mouse group were (185.44 ± 68.97) mm3,(83.91 ± 14.33) mm3,(123.70 ± 20.85) mm3,and (34.54 ± 10.72) mm3 respectively.From then on,the CDK2-shRNA mouse group,DTIC mouse group,and CDK2-shRNA + DTIC mouse group showed significantly decreased tumor growth rates compared with the control mouse group (F =11.819,P ＜ 0.05),and the tumor growth rate was significantly lower in the CDK2-shRNA + DTIC mouse group than in the DTIC mouse group (P =0.04).The calculated tumor growth inhibition rates in the CDK2-shRNA mouse group,DTIC mouse group and CDK2-shRNA + DTIC mouse group were 52.2％,41.2％ and 86.4％ respectively.Compared with the control mouse group,the CDK2-shRNA mouse group,DTIC mouse group,and CDK2-shRNA + DTIC mouse group showed significantly increased tumor cell apoptosis indice (32.93％ ± 3.72％,21.62％ ± 3.54％,63.29％ ± 4.74％ respectively;F =222.25,P ＜ 0.05).Moreover,the tumor cell apoptosis index was significantly higher in the CDK2-shRNA + DTIC mouse group than in the DTIC mouse group (P ＜ 0.01).Conclusion CDK2 gene silencing can enhance the inhibitory effect of DTIC on the growth of melanoma,and show a synergistic effect with DTIC,likely by increasing the apoptosis of tumor cells.

BACKGROUND:The Wnt/β-catenin signaling pathway is one of the most important signaling pathways in stem cel regulation, which is involved in regulation of cel proliferation and differentiation. OBJECTIVE:To investigate the expression of Wnt/β-catenin main signaling molecule in inflammatory bowel tissues treated with bone marrow mesenchymal stem cel transplantation. METHODS:2,4,6-Trinitrobenzene sulfonic acid was used for establishing inflammatory bowel diseases rat models. Bone marrow mesenchymal stem cels labeled with green fluorescent protein were transplanted into rat modelsviatail vein. Normal saline was injected as control. The expression of Wnt/β-catenin signaling molecule was detected in the large intestine tissue of inflammatory bowel disease rat models by quantitative RT-PCR at 14 and 28 days after transplantation. RESULTS AND CONCLUSION:Real-time quantitative PCR results showed that the expression of Wnt3a andβ-catenin in the inflammatory bowel tissue increased significantly (P < 0.05), while no difference in the expression of c-myc (P > 0.05). The expressions of Wnt3a, β-catenin and c-myc in the transplantation group were significantly lower than those in the control group after transplantation (P <0.05). These findings indicate that the Wnt/β-catenin signaling pathway plays important roles in inflammatory bowel disease and repair after bone marrow mesenchymal stem cel transplantation, while this pathway may promote stem cels differentiating into intestinal epithelium, promote recovery from inflammatory bowel disease, repair inflammatory area, and restore intestinal tissue homeostasis.

Objective To explore the characteristics of resting myocardial perfusion of type 2 diabetes mellitus(T2DM) by real-time myocardial contrast echocardiography(RT-MCE). Methods Thirteen patients with T2DM and 12 in control were enrolled in the study. RT-MCE was performed using a continuous infusion of SonoVue with vena mediana in elbow. Images of left ventricle filled with contrast were acquired from apical 4-, 2-chamber and long axis views with real-time myocardial contrast mode of GE Vivid 7 Dimension system. All above images were captured in continuous 3 cardiac cycles before "flash" ,and then 15 cardiac cycles after "flash". All clips were stored for off-line analysis. Results In quiescent condition, there were significant decrease on indices of A,k and A × k in patients compared with control group (A:6.46 ±1.60 vs 6.81 ± 1.53, P ＜0.05;k:1.04 ± 0.39 vs 1.28 ± 0.31*, P ＜0.01;A× k:6.55 ± 2.72 vs 8.78 ±3.16, P ＜ 0.01, respectively). Conclusions RT-MCE could evaluate the early change of myocardial perfusion in T2DM patients and offers important reference for clinical diagnosis and treatment.

Objective To study the relationship between level of plasma glucose and cognitive function in patients with Parkinson's disease.Methods Two hundred PD patients were assessed cognitive function using Mini-Mental State Examination (MMSE),Montreal Cognitive Assessment (MoCA),Wechsler Intelligence Scale and Wechsler Memory Scale.The patients were divided into cognitive normal group (n=91) and cognitive impairment group (n=109).One hundred twenty-six normal subjects were enrolled as control group (n=126).The levels of fasting plasma glucose (FPG),postprandial plasma glucose (2hPPG),glycosylated hemoglobin (HbAlc) and the prevalence of diabetes mellitus were compared among the groups.The effect of blood glucose level on the cognitive function of PD patients was analyzed by Binary Logistic Regression.Results The levels of FPG,HbAlc and the prevalence of diabetes mellitus [5.19 (0.72),5.7％ (0.5％),14％] were significantly higher than those in the normal control group [4.85(0.79),5.6％ (0.5％),6％] (P＜0.05).The levels of FPG in PD patients with cognitive impairment [5.21 (1.32)] was significantly higher than that in PD patients with cognitive normal group [4.81 (0.95)] (P＜0.05).Although 2hPPG and HbAlc increased slightly in PD patients with cognitive impairment,the difference did not reach an significant level (P＞0.05).Binary Logistic Regression analysis showed that FPG(OR:1.764;95％ CI:0.06-3.244;P=0.068) was not associated with the impaired cognitive function in PD patients.Conclusion The present study has not revealed an association between the incidence of cognitive impairment in patients with PD and plasma glucose level although high plasma glucose may be a high risk factor for PD patients.

Objective To explore the clinical value of color Doppler flow imaging(CDFI) combined with quantitative analysis technique to evaluate renal flow perflusion. Methods Twenty patients with different renal diseases matched with healthy control subjects were examined with Philips iU22 CDFI system. CDFI system and Qlab quantitative technique were used to measure renal restrictive index(RI), vascularization index(VI), flow index(FI), vascularization-flow index (VFI). The patients all received renal single-photon emission computed tomography (SPECT) examination. Results The RI of 32 kidney with lower flow perfusion in SPECT were higher than in the contol group (P <0.01). VI,FI, VFI were lower than those in control(VI, VFI, P <0.01 ;FI, P <0.05). Conelusious The renal blood flow perfusion of patients with renal diseases are lower than that of healthy people. It has high clinical value using CDFI combined with quantitative analysis technique to evaluate the renal perfusion of the patients with renal diseases.

Objective To assess the value of quantitative color Doppler sonography in patients with systemic lupus erythematosus (SLE). Methods The right renal in 21 patients with SLE and 22 normal subjects were examined with Philips IU22 ultrasonic diagnostic device. Resistence index (RI) of interlobar artery and interlobular artery were measured with color Doppler sonography, vascularity index (VI), flow index (FI) and vascularization flow index (VFI) which reflecting the renal cortical blood flow were calculated with quantitated analysis software. All the parameters between two groups were compared and analyzed. Results The color signals of renal cortical blood flow in patients with SLE were inferior to those of the normal group, especially in the area near the edge of renal. No statistical difference of RI was found between the two groups (P>0.05), while VI, FI and VFI in SLE group were significantly lower than those of the normal group (VI, VFI, P<0.01, FI, P=0.01). Conclusion Color Doppler sonography combined with blood flow quantitative technique provide a helpful, sensitive and convenient tool to detecte the renal changes in SLE patients.

Objective To observe the rotation of subendocardium and subepidium by two-dimensional speckle tracking imaging(2D-STI),and to evaluate its performance in diastolic heart failure patients(DHF)with a normal left ventricular ejection fraction. MethodsNinety-seven consecutive clinically stable patients were enrolled in this study [41 healthy controls,36 with diastolic heart failure,20 with systolic heart failure (SHF)]. High frame rate dynamic two-dimensional images were recorded at the left ventricular short-axis view,including basal, papillary muscle and apical planes. Subendocardial and subepicardial global rotation were measured using Q-lab 7.0 software offline. Results ① In all the subjects, the rotation of the subendocardium was obviously greater than that of subepicardium. ②As seen from the apex,left ventricular subendocardium and subepicardium performed a wringing motion with a clockwise rotation at the base and countclockwise rotation at the apex. ③In the apical plane, subendocardial rotation was significantly lower in both heart failure groups than in controls,and was depressed to a larger extent in SHF patients than in those with DHF. Subepicardial rotation was no significant difference between the DHF group and the control group, though it was significantly lower in patients with SHF. ④At the base, the rotation of subendocardium and subepicardium were not different between DHF and control groups, but it was significantly reduced in patients with SHF. Conclusions The subendocardial rotation is reduced, but subepicardial rotation is normal in DHF patients. On the other hand, in patients with SHF, subendocardial and subepicardial rotation are both reduced. The left ventricular systolic properties are impaired in DHF patients.

Objective:To explore the effects of dual task training assisted by a lower limb rehabilitation robot on lower extremity mobility and the walking ability of stroke survivors.Methods:Sixty-one stroke survivors were randomly divided into a control group and an experimental group with 30 in the control group and 31 in the experimental group. In addition to routine exercise training and physical therapy, both groups were given cognitive-motor dual task training 5 times a week for 3 weeks. But only in the experimental group was the dual task training assisted by a lower limb rehabilitation robot. Both groups′ lower limb motor function, walking ability, cognition, balance and ability in the activities of daily living were evaluated before and after the experiment using the Fugl-Meyer lower extremity assessment (FMA-LE), functional ambulation categories (FAC), the digital span test (DST), the Berg Balance Scale (BBS) and the Modified Barthel Index (MBI). Additionally, 6 survivors of a right hemisphere stroke from the experimental group received cognitive-motor dual task training both with and without the robotic assistance alternately. Near-infrared functional brain imaging was applied before and after the intervention, and the functional network connectivity of the resting brains was analyzed.Results:After the intervention the average FMA-LE, FAC, BBS and MBI scores had improved in both groups, with the improvement in the experimental group significantly better than in the control group on average. In terms of cognition there was no significant difference in the DST forward and backward assessment results between the two groups. The analysis of brain network functional connectivity showed that the intensity of functional connectivity between the left prefrontal cortex (PFC) and the left premotor cortex and supplementary motor cortex (PMC/SMA) increased significantly more, on average, after training assisted by the robot.Conclusion:Dual task training with the assistance of a lower limb rehabilitation robot can effectively improve the lower limb motor function, walking, balance and ability in the activities of daily living of stroke survivors. Enhanced functional connection of the PFC and the PMC/SMA in the healthy hemisphere induced by the robot may be the cause.