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.

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

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

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 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.

· AIM: To study the inhibitory effect of pyrrolidine dithiocarbamate (PDTC) on the inflammatory reaction in an experimental proliferative vitreoretinopathy (PVR)model with laser flare cell meter (LFCM).· METHODS: A total of 20 pigmented rabbits were divided into two groups randomly, with 10 rabbits in each group. After the creation of retinal holes, 0.1mL PDTC was injected intravitreally into the right eyes of Group 1(A1) and the left eyes of Group 1 (A2), and 0.1mL balanced saline solution (BSS) into the right eyes of Group2 (B1). One hour later, 0.1mL BSS into the eyes of A1,and 5000U IL-1 β in 0.1mL BSS was injected intravreally into the eyes of A2 and B1. Clinical evaluation and LFCM examination were performed before retinal injury (PO)and at 4h, 24h, 1, 2 and 4wk after the second injection(P4h, P24h, Plwk, P2wk and P4wk). Histopathologic and immunohistochemical examination were also performed at these time points.· RESULTS: PDTC could inhibit the inflammatory reaction obviously from P24h to P2wk. The eyes of A1 and A2 recovered earlier than those of B1. Although inflammatory reaction in the 3 groups resolved completely by the end of P2wk measured with the slit-lamp microscope,the eyes of the B1 still showed obvious aqueous flare judged by the LFCM compared with those of A1 and A2.Histopathologic and immunohistochemical examination showed that nuclear factor- κ B (NF- κ B) was activated by IL-1 β and the PDTC had inhibitory effect on it without obvious toxicity to retina.· CONCLUSION: Inflammatory reaction involves in the rabbit model of PVR induced by injecting intravitreally IL-1 β and the PDTC can relieve it significantly. The LFCM provides a new, sensitive, objective and noninvasive method to quantify the inflammatory reaction in the PVR model.

Objective To observe the effects of hIL-10-MSCs on heterologous T cell prolifera-tion. Methods hlL-10 was cloned by RT-PCR and then hIL-10/pLOX-cwGFPs was construct. The lentivirus was transfected into cavy MSCs and cell culture supernatant was tested for IL-10 protein by ELISA assay. The effects of hIL-10-MSCs on heterologous T cell proliferation were determined by CCK-8. The effect of peripheral blood mononuclearcell secretion IFN-γ was tested by EIASA. Results hIL-10/pLOX-cwGFP were constructed successfully and hIL-10-MSCs cell culture supernatant of IL-10 protein were increased obviously, hIL-10-MSCs could inhibit heterologous T cell proliferation sig-nificantly(P<0.05) but could not induce T cell proliferation (P<0.05). Adding IL-2 could reverse this inhibition(P<0.05), hIL-10-MSCs cell culture supernatant could inhibit peripheral blood mono-nuclearcell secretion IFN-γ. Conclusion hIL-10-MSCs may play an important role in significant im-munosuppression of heterologous T cell proliferation and suppression of secretion IFN-γ by peripheral blood mononuclear cells.