As the amount of PET/CT increases,the amount of cyclotron also increases.Hospitals are confronted with some similar problems in cyclotrons purchase.The article is about the stability,after service,yields of nuclide and the particle energy of cyclotron.The stability of cyclotron is the most important factor,and after service is in the next place,while the yields of nuclide are also very important.

Objective To compare the long-term efficacies of anterior cervical decompression and fusion (ACDF)and conservative therapy in treatment of unisegmental cervical disc herniation and to analyze the degree of the long-term adjacent segment disc degeneration in ACDF.Methods 120 patients treated in our hospital from January 2006 to January 2009 were selected,60 patients underwent ACDF as operation group and 60 patients underwent physical therapy and drug (conservative therapy)as non-operation group. All the patients were recorded when they were diagnosed with cervical unisegmental cervical disc herniation, and followed-up for 60 months, and they were evaluated with American Spinal Injury Association (ASIA)score,Visual Analogue Scale (VAS),Japanese Orthopedics Association (JOA) score and imaging (Miyazaki grading system). Results 107 patients were followed up,55 patients treated by operation were followed up for an average 59.7 months (59.7±0.4),while 52 patients treated by conservative therapy were followed up for an average 58.8 months (58.8±1.5).Compared with before treatment, the ASIA, JOA, and VAS scores of the patients in two group after treatment were improved (P<0.05),and the scores of ASIA,VAS,and JOA in operation group were superior to that in non-operation group (P<0.05),and adjacent segment disc degeneration occurred more frequently in the upper adjacent segment than the lower adjacent segment (P<0.05).Conclusion ACDF is superior to conservative therapy in alleviating symptoms and improving prognosis of unisegmental cervical disc herniation. Long-term postoperative disc degeneration mainly occurrs in the upper segment.

Objective To establish a simple and effective method for isolation and culture of mouse adipose-derived stem cells (mASCs)in vitro,in order to provide the sufficient sources of seed cells for the research of mesenchymal stem cells.Methods The mouse inguinal fat tissues were isolated in vitro and performed a digestion with 0.1% collagenase type NB4,then adipose-derived stem cells(ASCs)were seeded and adhered to the culture dishes in low glucose DMEM containing 10% fetal calf serum.The cellu-lar morphology,in vitro proliferation capacity,multidifferentiation potential and immunophenotype were assessed.Results The mASCs showed good cell morphology,extremely strong proliferation capacity and potential of adipogenesis,osteogenesis and chon-drogenesis via in vitro three-dimensional induction.The cellular surface antigen phenotype was consistent with that reported by lit-erature,and the expression of CD34 and CD105 was positive,Sca-1 was highly expressed,CD45 and SSEA-1 were not expressed. Conclusion Using the experimental methods in this research can culture the high purity of mASCs with the excellent stem cell properties and extremely strong proliferative ability.

Peer review anti assessment includes the evaluation of expert and index system in two parts.The current status of anti-assessment study mainly focused on expert,much of the index system anti-assessment study reports.As can be seen from the status,the anti-assessment system not yet formed and the application is rare.In urgent need of further research to improve our peer-reviewed scientific research.

BACKGROUND:Elderly patients with degenerative lumbar degeneration often appear insufficient holding power of pedicle screw in spine surgery, which is prone to occur de-pinning and leads to insecure fixation. How to increase the holding power of screws has become a hot research. OBJECTIVE:To observe the early clinical effect of pedicle screws with cement augmentation for treating lumbar degenerative diseases in elderly patients. METHODS:A total of 65 old patients with lumbar degenerative diseases received a treatment between August 2012 and April 2014, and were divided into two groups according to the treatment strategy:treatment group (n=24;internal fixation of pedicle screws with cement augmentation) and control group (n=41;routine internal fixation of pedicle screws). General conditions of patients in two groups were observed and compared. Visual analog scale (VAS) and Japanese Orthopaedic Association (JOA) score system were used for evaluating the lumbar and back pain, and restoration of neurological function in lower limbs respectively. RESULTS AND CONCLUSION:Al of the patients successful y received the surgery and then were fol owed up from 3 to 20 months. The anterioposterior and lateral X-ray film revealed no loosening, loss, fracture of the screws, and no loss of intervetebral space height was found. There was no significant difference in the blood loss and hospital stay between two groups (P>0.05). JOA at postoperative 3 and 6 months, and VAS score at postoperative 3 months were significantly improved after the treatment of pedicle screws with cement augmentation, when compared to control group (P<0.05). VAS scores showed no difference at 6 months postoperatively in two groups (P>0.05). Pedicle screws with cement augmentation for treating lumbar degenerative diseases have the advantages of improving the screws holding strength, reconstructing the stability of lumbar vertebra and obtaining clinical efficacy on degenerative spine.

Although two chain transfering separately could be used to overcome the volume limitation of adeno-associated virus vectors (AAV) in coagulation factor VIII (FVIII) gene delivery, it leads to chain imbalance for inefficient heavy chain secretion. In this study we aimed to improve the efficacy of two chain strategy in FVIII gene delivery through the degradation of glucose-regulated protein 78 (GRP78) known as a protein chaperone in endoplasmic reticulum (ER) by deoxynivalenol (DON) to decrease GRP78-bound FVIII heavy chain. By treating the two-chain gene transduced 293 cells with DON, the heavy chain (HC) secretion and FVIII bioactivity were observed. Data showed that 293 cells after three hours post-treatment with DON at a concentration of 500 ng mL(-1) resulted in obvious decrease the level of GRP78 but no effect on the cell proliferation. The HC secreted from DON-treated cells transfected with HC gene alone was 59 +/- 11 ng mL(-1), higher than that secreted by control cells (15 +/- 4 ng mL(-1)), and the HC secretion was further increasing to 146 +/- 34 ng mL(-1) in light chain (LC) gene co-transfected cells with an activity measured up to 0.66 +/- 0.15 U mL(-1), also greater than control cells (76 +/- 17 ng mL(-1) and 0.35 +/- 0.09 U mL(-1)). Taken together, these data suggest that DON-mediated GRP78 down-regulation could improve the efficacy of two-chain FVIII gene transfering by facilitating HC secretion, providing an experimental basis for in vivo dual-AAV application in FVIII gene delivery.

In our recent study by exploring an intein-based dual-vector to deliver a B-domain-deleted FVIII (BDD-FVIII) gene, it showed that covalently ligated intact BDD-FVIII molecules with a specific coagulant activity could be produced from expressed heavy and light chains by protein trans-splicing. Here, we assessed the hypothesis that the efficiency of trans-splicing may be increased by adding to the intein sequences a pair of leucine zippers that are known to bring about specific and strong protein binding. The intein-fused heavy and light chain genes were co-transferred into cultured COS-7 cells using a dual-vector system. After transient expression, the intracellular BDD-FVIII splicing was observed and the spliced BDD-FVIII and bioactivity secreted to culture media were quantitatively analyzed. An enhanced splicing of BDD-FVIII with decreased protein precursors from gene co-transfected cells was observed by Western blotting. The amount of spliced BDD-FVIII and bioactivity secreted to the culture media were 106 +/- 12 ng x mL(-1) and 0.89 +/- 0.11 U x mL(-1) analyzed by ELISA and Coatest method respectively, which was greater than leucine zipper free intein-fused heavy and light chain genes co-transfected cells (72 +/- 10 ng x mL(-1) and 0.62 +/- 0.07 U x mL(-1)). The activity of cellular mechanism-independent protein splicing was also improved, as showed by the increasing of spliced BDD-FVIII and bioactivity in culture media from combined cells separately transfected with heavy and light chain genes which was 36 +/- 11 ng x mL(-1) and 0.28 +/- 0.09 U x mL(-1). It demonstrated that the leucine zippers could be used to increase the efficiency of protein trans-splicing to improve the efficacy of a dual-vector mediated BDD-FVIII gene delivery by strengthening the interaction between the two intein-pieces fused to heavy and light chains. It provided evidence for further study in animal model using a dual-adeno-associated virus vector to deliver FVIII gene in vivo.

As synthesized by vascular endothelial cells and megakaryocytes, the von Willebrand factor (vWF) plays an important hemostatic role in the binding to and stabilizing blood coagulation factor VIII (FVIII) and preventing its enzymatic degradation. Our recent work demonstrated intein can efficiently ligate BDD-FVIII (B-domaim deleted FVIII) posttranslationally by protein trans-splicing after transfer of split BDD-FVIII gene by a dual-vector system. In this study we investigated the effect of vWF on secretion and activity of intein-ligated BDD-FVIII. We observed the levels of full-length BDD-FVIII antigen secreted into culture supernatant by ELISA and their activity by Coatest assay after transfection of cultured 293 cells with intein-fused BDD-FVIII heavy- and light-chain genes simultaneously with the vWF gene co-transfected. The data showed that the amount of full-length BDD-FVIII protein and their bioactivity in vWF gene co-transfected cell supernatant were 235 +/- 21 ng x mL(-1) and 1.98 +/- 0.2 u x mL(-1), respectively, greater than that of non-vWF co-transfected cell (110 +/- 18) ng x mL(-1) and 1.10 +/- 0.15 u x nL(-1)) or just BDD-FVIII gene transfected control cell (131 +/- 25 ng x mL(-1) and 1.22 +/- 0.18 u x mL(-1)) indicating the benefit of vWF gene co-transfection in the secretion and activity of intein-spliced BDD-FVIII protein. It provided evidence that vWF gene co-transfer may be useful to improve efficacy of gene therapy for hemophilia A in protein splicing-based split FVIII gene transfer.

This study is to construct a chimeric human/porcine BDD-FVIII (BDD-hpFVIII) containing the substituted porcine A1 and A3 domains which proved to have a pro-secretory function. By exploring Ssp DnaB intein's protein trans-splicing a dual-vector was adopted to co-transfer the chimeric BDD-hpFVIII gene into cultured COS-7 cell to observe the intracellular BDD-hpFVIII splicing by Western blotting and secretion of spliced chimeric BDD-hp FVIII protein and bio-activity using ELISA and Coatest assay, respectively. The dada showed that an obvious protein band of spliced BDD-hpFVIII can be seen, and the amount of spliced BDD-hpFVIII protein and bio-activity in the supernatant were up to (340 +/- 64) ng x mL(-1) and (2.52 +/- 0.32) u x mL(-1) secreted by co-transfected cells which were significantly higher than that of dual-vector-mediated human BDD-FVIII gene co-transfection cells [(93 +/- 22) ng x mL(-1), (0.72 +/- 0.13) u x mL(-1)]. Furthermore, a spliced BDD-hpFVIII protein and activity can be detected in supernatant from combined cells separately transfected with intein-fused BDD-hpFVIII heavy and light chain genes indicating that intein-mediated BDD-hpFVIII splicing occurs independently of cellular mechanism. It provided evidence for enhancing FVIII secretion in the research of animal models using intein-based dual vector for the delivery of the BDD-hpFVIII gene.

Objective To develop the dual-beam technology of cyclotron to produce radioactive nuclide, which can increase radiopharmaceuticals output. Methods A new target is installed in cyclotron standby target, and at the same time the tubes and electronic components are fixed to produce radioactive nuclide through dual-beam technology, the product is tested by the dose calibrator. Results The radioactive nuclide can be produced through dual-beam technology and the output is nearly double times about which a single target. Conclusion The productions of cyclotron radioactive isotope are improved by dual-beam technology and meet the clinical needs of PET/CT examination for drugs.