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

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

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.

Objective To evaluate the outcome of unconstrained shoulder arthroplasty for severe injury of proximal part of the humerus.Methods Twelve eases were all treated with unconstrained shoul- der arthroplasty.Six patients with complex fractures of proximal humerus and four with bone tumor in the proximal part of the humerus used hemiarthroplasty and two patients with osteoarthritis were managed with total shoulder arthroplasty.Cemented prostheses were used in all the cases.Results The average age was 65 years and the follow-up was 2.3 years.Two cases of complex fractures had light pain with limited external rotation of the shoulder.No pain or prosthetic stems loosening were found,and the range of motion and the function of the shoulder were satisfactory in other cases.Conclusion Unconstrained shoulder arthroplasty is a satisfactory and safe technique for severe injury of proximal humerus.

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.

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.