Objective To construct lentiviral vector miR30-CD133 targeting CD133 gene and investigate its effects on SMMC7721 cells. Methods Thespecific sequence of DNA which containing both sense and antisense Oligo DNA of the targeting sequence were designed, synthesized and cloned into the pPRIME vector. pPRIME-miR30-CD133, psPAX2 and pMD2G were co-transfected on 293T cells to get the lentivirus the transfection efficiency was investigated by confocal laser scanning microscope. The expression of miR30-CD133 on CD133 were detected by qRT-PCR and Western blott. The proliferation and apoptosis effect of miR30-CD133 was respectively evaluated by CCK-8 assay and AnnexinV/PI. Results Functional PFU titers of PRIME-miR30-CD133 were 6.58 × 109 PFU/mL. The expression of CD133 mRNA in sh CD133 group (0.18 ± 0.04) was less than Blank group and shNC group (P<0.05). The expression of CD133 protein in sh CD133 group was less than Blank group (10%) and shNC group (8%) (P < 0.05). Cells proliferation activity were significantly inhibited in shCD133 group compared with control group. Apoptosis rate were significantly increased in shCD133 group (41.3%) compared with Blank group (25.3%)and shNC group (24.3%). Conclusion The lentivirus miR30 vector targeting CD133 gene was successfully constructed, which will be a basis to the further CD133 gene studies.

Objective To study the effects of lentiviral vector of microRNA targeting IGF1R gene on growth of liver cancer cells.Methods The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pPRIME vector,named pPRIME-IGFIR-miR30-shRNA.The viruses were propagated on 293T cells.Viruses were purified by CsCI gradient according to standard techniques,and functional PFU titers were determined by plaque assay on 293 cells.The effect of pPRIME-IGFIR-miR30-shRNA on IGFIR expression of Hep3B、SMMC7721 cells was detected by RT-PCR and Western blot.The antitumor potential of PRIME-IGFIR-miR30-shRNA to Hep3B、SMMC7721 cells was evaluated by CCK-8 assay and Tunel.Results PRIME-IGFIR-miR30-shRNA was constructed successfully.Functional PFU titers of pPRIME-IGFIR-miR30-shRNA were 4.58?109PFU/ml.PRIME-IGFIR-miR30-shRNA inhibited IGFIR expression and the proliferation of Hep3B、SMMC7721 cells.Conclusions PRIME-IGFIR-miR30-shRNA expressing IGFIR-siRNA can inhibit IGFIR expression and may be used for further investigation of gene therapy of liver cancer.

Adult ; Breast Neoplasms ; diagnostic imaging ; radiotherapy ; surgery ; Cone-Beam Computed Tomography ; Female ; Humans ; Mastectomy, Segmental ; Postoperative Period ; Radiotherapy Planning, Computer-Assisted ; methods ; Respiration

Air Pollution ; analysis ; Barium ; analysis ; chemistry ; Solubility ; Spectrophotometry, Atomic ; methods ; Workplace

Objective To study the therapeutic effects and mechanisms of Astragaloside-Ⅳ (ASG-Ⅳ) on diabetic cardiomyopathy (DCM) in the rat diabetic model. Methods Forty SD(Sprague-Dawley)healthy rats were used. The diabetic retinopathy rats model were induced by STZ, 45 mg/kg, 3d. Another 10 were injected the same amount of citrate buffer as a control group. Fasting blood glucose was measured with SureStep Plus detector 72 h later. The blood glucose of the diabetes model was ≥16.7 mmol/L. And 12 weeks later, DCM rats were divided into 4 groups randomly in the experiment, includes: DCM, ASG-Ⅳ-L (10 mg/kg), ASG-Ⅳ-M (30 mg/kg), ASG-Ⅳ-H (60 mg/kg)groups. After give dugs 4 weeks, the phosphokinase isoenzyme (CK-MB) and LDH level were tested, the cardiac hypertrophy was evaluated by HW/BW and LVW/BW. Activity of Na+-K+-ATPase and Ca2+-ATPase were determined in left ventricular tissues by ATPase ELISA Assay Kit. The content of FFA was measured and myocardial pathological examination was performed. Results Compared with the control group, blood and urine glucose were higher than experimental animal in diabetic model group, were significantly increased (P<0.05). LDH and Phosphokinase isoenzyme (CK-MB) level were significantly increased, the HWI and LVWI ratio were enhanced in DCM group (P<0.05). ASG-Ⅳ could reduce the ratio of HWI and LVWI, decrease the level of CK-MB and LDH, improve the pathomorphological changing of DCM rat model (P<0.05). Moreover, compared with DCM group, ASG-Ⅳ could restore the Na+-K+-ATPase and Ca2+-ATPase activity, reduced the content of FFA (P<0.05). Conclusion ASG-Ⅳ plays therapeutic effect on diabetic cardiomyopathy might via restore the Na+-K+-ATPase and Ca2+-ATPase activity, reduce the content of FFA, protect the myocardial energy metabolism in DCM.

Breast Neoplasms ; diagnostic imaging ; radiotherapy ; Female ; Humans ; Imaging, Three-Dimensional ; Mastectomy, Segmental ; Postoperative Period ; Radiotherapy Planning, Computer-Assisted ; methods ; Respiration ; Surgical Instruments ; Tomography, X-Ray Computed ; methods ; Tumor Burden

Humans ; Imaging, Three-Dimensional ; Neoplasms ; radiotherapy ; surgery ; Radiosurgery ; instrumentation ; methods ; Tomography, X-Ray Computed

Objective To observe the influence of hIL-10-MSCs on rejection of rat discordant liver xenotransplantation. Methods The model of cavia to rat discordant liver xenotransplantation was established. The rats were randomized into the control group, MSCs group, and hIL-10-MSCs group.The survival time and liver function of each recipient were observed and expression levels of E-Selectin, LFA-1, VCAM-1, and NF-κB determined by RT-PCR and ELISA 24 h after transplantation.Results The survival time was prolonged, liver function improved, and expressions levels of E-Selectin, LFA-1, VCAM-1 and NF-κB were significantly decreased in the hIL-10-MSCs group (P＜0.05).Conclusion hIL-10-MSCs are able to protect transplanted liver and decrease the expression of E-Selectin, LFA-1, VCAM-1, and NF-κB in the liver.

Objective To investigate the effects of alcohol on hippocampal apoptosis-inducing factor (AIF) and caspase-3 expression in cercbral ischemia/reperfusion in rats.Methods Sixty-eight healthy adult male Wistar rats were randomly divided into a sham operation group (n =4) and a saline control group as well as low-dose (1.0 g/kg),medium-dose (1.5 g/kg) and high-dose (2.0 g/kg) ethanol groups.The saline control group and each dose alcohol group were redivided into ischemia/reperfusion 0,1,2 and 3 h subgroups according to the intervention time points (n =4 in each group).A model of middle cerebral artery ischemia/reperfusion in rats was induced by the suture method.The corresponding doses of ethanol or an equal volume of saline were injected intraperitoneally at ischemia for 2 h and reperfusion for 0,1,2 and 3 h in the alcohol groups and the saline control group.At ischemia for 2 hand reperfusion for 24 h,the neurological deficit in rats was evaluated by using behavioral score.Immunohistochemistry assay was used to detect the expressions of AIF and caspase3 in the hippocampus of ischemic sides at ischemia for 2 hand reperfusion for 24 h.Results The behavior evaluation showed that the neurological deficit score was 0 in the sham operation group.The neurological deficit scores in the different dose ethanol groups were significantly lower than those in the saline control group at the same intervention time points (all P=0.000),and there were significant differences between the same intervention time points in the different dose ethanol groups (all P =0.000).The high-dose ethanol group was the lowest.There were no significant differences between the different intervention time points in the same dose ethanol groups (all P＞0.05).Immunohistochemistry revealed that the numbers of positive cells of AIF and caspase-3 in the sham operation group were 17.21 ±2.86 and 20.60 ±4.39,respectively,and they were significantly less than those in the saline control group and the each dose ethanol group at ischemia/reperfusion 0 h (all P =0.000); the number of positive cells of hippocampal AIF and caspase-3 in the different dose ethanol groups were all significantly less than those in the saline control group at the same intervention time points (all P =0.000).There were significant differences between the same intervention time points in the different dose ethanol groups (all P =0.000),and the high-does ethanol group was the lowest.There were no significant differences between the different intervention time points in the same dose ethanol groups (all P =0.000).Conclusions Alcohol has protective effect on the cerebral tissue in ischemia/reperfusion injury in rats.Its mechanism may be associated with the inhibition of AIF and caspase-3 expression.

NAD +-dependent cytosolic glycerol-3-phosphate dehydrogenase in Sacch aromyces cerevisiae is one of the key enzymes in metabolic pathway of glycerol . catalysing the reduction of dihydroxyacetone phosphate to glycerol-3-phosph ate.It has two isoenzymes.To study the differences between their structures, their expression of encoding genes and their functions may help increase the understan ding of the cell response mechanism to the hyperosmotic and anoxic conditions. In this paper the research on the two isoenzymes was reviewed.