Objective To investigate the impact of stable expression of perlecan shRNA lentiviral particles on proliferation of NIH3T3 cells. Methods Mouse fibroblasts were cultured.Lentiviral particles-green fluorescent protein (LV-GFP) was used to transfect the cultured NIH3T3 cells with multiplicity of infection (MOI) of 10,30 and 50.The GFP expression was observed with fluorescence microscopy after transfection for one week to estimate the proper MOI and the time of GFP expression needed.The transfection efficiency of LV-GFP with the proper MOI by fluorescence-activated cell sorting was detected.The stably transfected cell lines were developed by puromycin screening for more than 2 weeks.The third generation HFF in good condition was randomly divided into 3 groups:GFP group,shRNA group and control group.RT-PCR,Western blot and MTT assays were used to detect the expressions of perlecan mRNA and protein and cell proliferation in the 3 groups. Results Perlecan mRNA and protein showed high expressions in the control and GFP groups but low expressions in the shRNA group,with significant differences respectively between the shRNA group and the other 2 groups ( P ＜ 0.05).There was no significant difference between the 3 groups in the optical density at the first 2 days ( P ＞ 0.05).On 3 to 6 days the cells in the control and GFP groups grew normally while the cells in the shRNA group proliferated in a weak manner.the transfected cells in the shRNA group showed a significantly reduced proliferation rate compared with the other 2 groups ( P ＜ 0.05 ). Conclusion The growth of NIH3T3 cells can be inhibited significantly by transfection with perlecan shRNA lentiviral particles.

Objective To observe whether peripheral blood mononuclear cells (PBMCs) from patients with active systemic lupus erythematosus (SLE) can induce the adhesion of T lymphocytes to endothelial cells. Methods Sera and PBMCs were obtained from patients with active SLE and normal human controls. PBMCs were cultivated and culture supematants were harvested. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro with or without the presence of the sera or culture super-natants of PBMCs. Some cells were pretreated with the antibody to IL-17 before the treatment with the sera or supematants. After another 48-hour culture, RT-PCR and real-time PCR were used to detect the mRNA expressions of adhesion molecules, including intercellular adhesion moleeule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-cadherin in HUVECs, wound healing assay to estimate the motility of HUVECs. Additionally, T lymphocytes were added to HUVECs 48 hours after stimulation with the sera or supematants, the adhering of T lymphocytes to HUVECs was observed by microscopy. Results After stimulation with supematants of PBMCs from patients with active SLE, the mRNA expressions of ICAM-1, VCAM-1 and E-cadherin were significantly increased in HUVEC, while the increase could be inhibited by the antibody to IL-17. The elevation of adhesion molecule expression subsequently promoted the motility of HUVEC, mediated the adhesion of T lymphocytes to HUVEC, and the antibody to IL-17 could suppress the adhesion of T lymphocytes and motility of HUVEC. Conclusion The culture supematants of PBMCs from patients with active SLE can induce the expression of vascular cell adherin molecules and promote the adherin of T lymphocytes, which may in turn mediate the development of lupus vasculitis.

Objective To investigate influence of laser positioning system to position repeatability of patients treated with radiotherapy for pelvic tumors.Methods Forty patients with pelvic tumors were divided into two groups,using laser positioning system positioning (20 cases) and conventional positioning (20 cases),respectively,CBCT was used to verify and analyze the setup errors of two groups.Results The setup errors on X,Y and Z axes in laser positioning system positioning group were (2.63±1.07) mm,(2.38±1.28) mm and (2.46±1.21) mm,they were (3.04±1.22) mm,(2.55±1.35) mm and (3.09±1.37) mm in conventional positioning group.It has significance reduce on X and Z axes (P =0.040,0.003,respectively) by using laser positioning system,and has no significance on Y axis (P =0.432).Conclusion It can improve patient positioning repeatability effectively by using laser positioning system for patient positioning.

Objective To observe the effects of Dl-3-n-butylphthalide ( NBP) on the mitochondria infarction, size of myocardial infarction and myocardial apoptosis after acute myocardial ischemia in rats. Methods 92 male SD rats were divided into sham operation group (8 rats) , model group (21 rats) , and low-dose NBP group (21 rats) , medium-dose NBP group (21 rats) , high-dose NBP group (21 rats) . The model and NBP groups were made into MI model by ligation of the left anterior descending ( LAD) coronary artery, but not in sham-operated group. Model group and NBP group were taken heart specimens after coronary artery ligation. Cardiomyocyte apoptosis was ana-lyzed by TUNEL in each group. Size of MI was analyzed by TTC staining in sham-operated group, model group and high-dose NBP group. Electron perspective microscopy was applicated in observing mitochondria infarction in model group and high-dose NBP group after myocardial infarction. The expressions of Bcl-2 protein and Bax protein were detected by Western blot. Results Compared with model group, butylphthalide significantly increased expression of Bcl-2 protein ( P <0.05 ) and the ratio of Bcl-2/Bax ( P <0.05 ) , inhibited mitochondria infarction ( P <0.05 ) , reduced myocardial infarct size ( P<0.01 ) and cardiomyocyte apoptosis ( P<0.05 ) . Conclusion Bu-tylphthalide significantly inhibits myocardial infarction by increasing expression of Bcl-2 protein and the ratio of Bcl-2/Bax and decreasing mitochondria infarction, reducing myocardial infarct size and cardiomyocyte apoptosis in rats during the acute myocardial ischemia process.

Objective To explore the inhibitory effect of aprotinin on general inflammatory reaction in cardiopulmonary bypass(CPB),Methods 16 patients with cardiac valves substitution were equal divided at random two groups;control group and aprotinin group.Before the CPB operation,40min during CPB period,30min,4h after CPB termination,the integrin CD 11b expression of neutrophilic granulocytes in heparinization arterial blood was determined by using flow cytometer.Results 30min,4h after CPB operation termination,the CD 11b expression in control group obviously higher than those of preopreration(P

Objective To analyze the influence of different types of seizures and nonepileptic events on peripheral white blood cell (WBC)counts. Methods We prospectively collected blood samples from all patients and detered WBC count to evaluate the relation of each type of seizure,duration,frequencies,and ttme lapse between a seizure and collection of blood sample to peripheral WBC count. Results Peripheral WBC count was elevated in about 50% cases after a generalized seizure. The length of a seizures was positively associated, whereas its frequencies and the lapse time was negatively correlated with the increase in WBC counts. Conclusion Peripheral WBC counts increase significantly after a generalized seizure.

Objective: To investigate the growth inhibition effect of caffeic acid and DDP (cisplatin) on human lung adenocarcinoma A549 cells in vitro, and to explore the possible mechanism. Methods: A549 cells were cultured in vitro. The effect of caffeic acid alone or in combination with DDP on proliferation of A549 cells was detected by MTT assay. The effect of caffeic acid alone or in combination with DDP on apoptosis of A549 cells was detected by Hoechst33258 immunofluorescence staining and flow cytometry. The expressions of Bcl-2, Bax and caspase-3 proteins were detected by Western blotting. Results: Caffeic acid or DDP alone or in combination could significantly inhibit the proliferation and induce apoptosis of A549 cells, and the combination of these two drugs had a synergistic effect. Western blotting result showed that the combination of caffeic acid and DDP synergistically inhibited the expression of Bcl-2 protein and enhanced the expression of Bax protein and the activation of caspase-3. Conclusion: Both caffeic acid and cisplatin may improve the growth-inhibitory and apoptotic activities of human lung cancer A549 cells in vitro. These effects are enhanced with the combination of caffeic acid and DDP in a synergistic manner. Copyright © 2012 by TUMOR.

Objective To evaluate the application of improved belly board in postoperative patients of rectal cancer radiotherapy and explore the influence of its set-up repeatability.Methods CBCT was used to measure the intra fractional and inter fractional setup errors using normal or improved belly board respectively.The data was analyzed with statistic method.Results There was no significance of intra fractional setup errors on x-axis (P ＞ 0.05).There was significance on y and z-axis (P ＜ 0.05).There was no significance of interfrational setup errors on x-axis (P ＞ 0.05) and there was significance on y and z-axis (P ＜ 0.05) using normal belly board.There was no significance of intrafractional setup errors on x,y and z-axis (P ＞ 0.05).There was also no significance of interfractional setup errors on x,y and z-axis (P ＞ 0.05) with using improved belly board.Conclusion The method of improved belly board has more advantage than nomal belly board in controlling set-up repeatability,and it is conductive to improve accuracy of patients treatment.

Objective: To investigate the effects of oral Lactobacillus Plantarum supplementation on apoptosis of liver and tight junction of hepatocytes in an experimental model of obstructive jaundice.Methods: Wistar rats were randomly divided into five groups: sham-operated;bile duct ligation;bile duct ligation +LP(0.2 g/5 mL/day);bile internal drainage;bile internal drainage+LP.Hepatocyte apoptosis was quantitated using TUNEL assay.Transmission electron microscopy,RT-PCR and Western blot were used to observe tight junction associated proteins.Results: Compared to the sham-operated,a significant increase in apoptosis(P

Objective To make a further understanding of mammographic features of simple cyst of the breast (SCB).Methods Molybdenum target radiographic signs in 39 cases with SCB proved pathologically and were retrospectively analyzed. Results Of the 56 SCB in 39 cases, 23 were diagnosed rightly as SCB, 10 as fibroadenoma, 2 as cancer and the remaining 4 were undefined. The diagnostic accuracy and misdiagnosis were 59.0% and 41.0% respectively. Radiograph showed round in 12, ovoid in 38, mild lobed in 4 and comet tail form in 2. The borders of cyst were distinct and sharp in 26, distinct partly and indistinct partly in 27,indistinct completely in 3. The density was homogeneous in 54 with calcifications in 2. Of the 56 SCB, the length axes of cyst were upright to chest wall in 20 and towards nipple in 18. The cysts could be transformed with pressure in 4.Conclusion Careful analysis of radiographic features of SCB is helpful for diagnosing accurately this disease.