ce, sIMRT and IMRT radiotherapy techniques can protect the lung and spinal cord well.

Objective To summary the clinical applied value of interventional therapy in hepatic venous outflow obstruction after orthotopic liver transplantation(OLT).Methods The clinical data of 27 patients suspect with hepatic outflow obstruction out of OLT patients were analyzed retrospec- tively.Most of them presented with liver dysfunction,like ascites,jaundice,or hydropsia of lower ex- tremity as BCS.These patients accepted venography and endovascular treatment if venous outflow ob- struction was found.Results By venography,one case of thrombus in inferior vena cava,one case of hepatic vein stenosis,13 cases of inferior vena cava stenosis(3 cases were associated with hepatic vein stenoses)were identified.Stent implantation was successfully performed on 10 patients,and balloon angioplasty on 4 patients.Rapid,dramatic resolution of symptoms was achieved in those patients. Hepatic vein restenosis occurred in one case 8 months after balloon dilatation,and treated with stent implantation.Inferior vena cava restenosis occurred in one case 2 years after balloon dilation,and trea- ted with another balloon expanding.Patients remained completely asymptomatic at 4 months to 5 years of follow-up.Conclusion The venacavographic balloon angioplasty and metallic stent replacement are safe and useful for post-OLT with venous outflow obstruction.

Adenocarcinoma ; genetics ; Adult ; Age Factors ; Aged ; Base Sequence ; Carcinoma, Squamous Cell ; genetics ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Sequence Deletion ; Smoking ; genetics

OBJECTIVETo study the effect of Hath1 gene transfer on the proliferation of colonic cancer cells in vitro.

METHODSThe recombinant plasmid pcDNA3.1(+)-Hath1 was transfected into HT29 colonic cancer cells, and 3 positive cell clones were randomly selected to test the levels of Hath1 mRNA, Muc2 mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time RT-PCR and Western blotting. The proliferation of the transfected HT29 cells was observed by means of colony formation assay and xenograft growth in nude mice.

RESULTSHath1 significantly down-regulated of cyclin D1 and up-regulate of p27 expressions and inhibited the proliferation of HT29 cells.

CONCLUSIONHath1 gene may be an anti-oncogene in colon carcinogenesis.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; pathology ; Cyclin D1 ; genetics ; metabolism ; Gene Transfer Techniques ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mucin-2 ; genetics ; metabolism ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics

Objective To investigate the ostengenie potential of adipose-derived stem cells(AD- SCs)when exposed to adenovirns containing hBMP-2 cDNA(Adv-hBMP-2)and offer a choice of cell source for gene therapy and tissue engineering.Methods Human adipose tissues were obtained from patients who received orthopaedic surgery or liposuction.ADSCs were obtained by digesting the adipose tissues.Firstly,flowcytometric analysis was performed for the confirmation of mesenchymal stem cell ori- gin and the surface markers including CD34,CD44,CD68,CD71,CD90,and CD105.The ADSCs were transfected by Adv-hBMP-2 and the effects were tested in vitro,lmmunoprecipitation and Western blotting and ELISA were performed for confirming BMP gone transduction and its stable expression.The transform of ADSCs was assessed by extracellular ALP staining,intracellular ALP spectrophotometry,von Kossa staining and RT-PCR.In the in vivo experiment ADSC-Adv-hBMP-2 cells were injected into the hind limb of nude mice and analyzed radiographically and histologically.Results ADSCs were successfully isolated from human adipose tissues.The isolated ADSCs expressed CD44,CD71,CD90 and CD105 and CD34 and CD68 were absent.The result confirmed the mesenchymal stem cell origin of the cells.West- ern blotting and ELISA confirmed successful and persistent hBMP-2 production by ADSC-Adv-hBMP-2 cells.Extracellular ALP staining,intracellular ALP spectrophotometry,yon Kossa staining and RT-PCR revealed that ADSCs treated with Adv-hBMP-2 had a tendency of transfering into osteoblast.X-ray and H&E sections from hind limb of nude mice injected with ADSC-Adv-hBMP-2 cells confirmed bone forma- tion at 2 weeks.Conclusions Liposuction aspirates contain abundant ADSCs that can be transduced with hBMP-2 gene,and the tranduced ADSCs differentiate into the osteoblast.ADSCs may be an ideal source of mesenchyme-lineage stem cells for gone therapy and tissue engineering.

Objective: To investigate the relationship between the image quality of 4D cone beam CT (4D-CBCT) and the characteristic parameters of respiratory motion. Methods: Catphan 500 was fixed on the Mapcheck XY/4D table by a fixation device. The control platform was utilized to simulate the static and 19 motion states. A synergy accelerator was adopted to perform 4D-CBCT scan of the image quality detection module under different motion states. By analyzing the geometric accuracy, spatial resolution, low-contrast visibility, CT value uniformity and motion position accuracy of the images of the 10 phases of the reconstructed images, the relationship among the image quality of 4D-CBCT, motion direction, amplitude and cycle was analyzed. Results: 4D-CBCT images could not be properly reconstructed in the static state or only in the left-right (LR) direction. The image quality was significantly worse than that in the SI direction. The contrast-to-noise ratio (CNR) was decreased as the motion cycle was increased. When the cycle was increased from 2 s to 6 s, the CNR was significantly declined from 3.33 to 1.72(P<0.01), whereas respiratory motion exerted no significant effect upon the geometric accuracy, spatial resolution and sagittal position. Conclusions The CNR, horizontal and vertical geometric accuracy and motion position accuracy of 4D-CBCT image quality are significantly affected by respiratory motion, whereas spatial resolution and sagittal geometric accuracy are not significantly influenced by respiratory motion.4D-CBCT requires the intervention of periodical motion in the SI direction. Excessively slow or absence of periodical motion can influence the image quality and even the images cannot be accurately reconstructed.4D-CBCT image reconstruction equally requires the intervention of periodical motion in the SI direction. The image quality is simultaneously affected by the amplitude and cycle of respiratory motion. For 4D-CBCT scan, the respiratory cycle of<6 s and an amplitude of>0.75 cm are recommended.

OBJECTIVETo study the cost of the hypertensive outpatients.

METHODSThe study randomly selected 460 insured patients with hypertension and investigated their cost on each case in the out-patient department through 2002, based on the electronic system of medical insurance.

RESULTSAs a whole, the distribution of hypertensive outpatient expenditure takes on the positively skewed, with the median of 1 567.9 Yuan RMB. With the increase of age, the average expenses in each age group increased accordingly. In the study, the average number of outpatient attendances per patient was 19.5, the average expenses per visit was 115.4 Yuan RMB. In age groups 40 - 49 and 50 - 59, expenses of outpatient in male and female groups are obviously different in 2002 (Wilcoxon W(40 - 49) = 36, P(40 - 49) = 0.037; Wilcoxon W(50 - 59) = 374, P(50 - 59) = 0.023), as well as the number of out-patients (Wilcoxon W(40 - 49) = 52.5, P(40 - 49) = 0.007; Wilcoxon W(50 - 59) = 379, P(50 - 59) = 0.028). When considering the factors of gender and age at one time, the outpatient expenditures in the male group were significantly different between the different age groups (chi(2) = 22.3, P < 0.001), as well as the number of outpatients (chi(2) = 25.4, P < 0.001). In addition, the expenditure of drugs, which took a large proportion of the total expenditure of hypertensive outpatients (about 83.6 percent), was divided into three parts according to the degree of correlation with hypertension: direct expenses related to the with disease, the indirect expenses and the irrespective. The proportions of each part were 19.9 percent, 32.3 percent and 47.8 percent respectively.

CONCLUSIONWhen economic evaluation of community prevention is carried out, the cost and cost-benefit analysis based on the analysis of outpatient expenditure and the proportion of expenses on hypertension should be taken into account. Additionally, to provide appropriate mode of medicare, to impact the behaviors and expenditure of patients, and to provide low-cost but good effective drug are also essential and important factors.

Adult ; Age Factors ; Aged ; Aged, 80 and over ; Ambulatory Care ; China ; Female ; Health Care Costs ; statistics & numerical data ; Hospitals, Community ; economics ; statistics & numerical data ; Humans ; Hypertension ; economics ; therapy ; Male ; Middle Aged ; Outpatients ; statistics & numerical data ; Sex Factors ; Time Factors

OBJECTIVETo summarize our experience in adult-to-infant living donor liver transplantation (A-ILDLT) and to analyze the efficacy and complications of A-ILDLT.

METHODSThe clinical data, surgical strategies and complications of 28 adult donors and infantile recipients who underwent A-ILDLT from April 2006 to December 2009 were retrospectively analyzed. These 28 patients (14 boys and 14 girls) aged from 80 days to 11.5 months with body weights of 3.08 to 10.3 kg at the time of operation . They suffered from biliary atresia with decompensated cirrhosis. The living donors were 15 mothers, 9 fathers, 3 grandma and 1 elder brother with ABO compatible with the infantile recipients. 27 Donor organs were the left lateral lobe grafts (segment II, III) and 1 graft was segment II. All patients were followed up for 5 to 24 months.

RESULTSThese grafts were orthotopically transplanted into the infantile recipients. The average length of stay was 9.3 days for the donor group without any complications. Postoperative immunosuppression included prednisone, Cyclosporin and mycophenolate mofetil (MMF). A total of 24 postoperative complications occurred in 20 recipients, including 5 vascular complications, 4 bleeding, 7 pneumonia, 2 bowel obstruction, 4 intestinal perforation and 3 rejection. Three recipients died of hepatic arterial thrombosis (HAT). The perioperative mortality rate of recipients was 10.7% (3/28) and the survival rate was 89.3% in peroperative period. One died of stricture of hepatic vein and 1 of accidental asphyxia during follow-up term. At present, 23 cases are still alive.

CONCLUSIONA-ILDLT has become an effective method to infants with end-stage liver disease. The postoperative vascular complication is the predominant cause of death.

Female ; Humans ; Infant ; Liver Diseases ; surgery ; Liver Transplantation ; methods ; Living Donors ; Male ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo evaluate the osteogenic potential of bone morphogenetic protein (BMP)-2 gene transfected goat bone marrow-derived mesenchymal stem cells (MSCs).

METHODSGoat bone marrow-derived MSCs were transfected by Adv-human bone morphogenetic protein (hBMP)-2 gene (Group 1), Adv-beta gal transfected MSCs (Group 2) and uninfected MSCs (Group 3). Western blot analysis, alkaline phosphatase staining, Von Kossa staining and transmission electron microscopy were adopted to determine the phenotype of MSCs. Then the cells were injected into thigh muscles of the nude mice. Radiographical and histological evaluations were performed at different intervals.

RESULTSOnly Adv-hBMP-2 transfected MSCs produced hBMP-2. These cells were positive for alkaline phosphatase staining at the 12th day and were positive for Von Kossa staining at the 16th day after gene transfer. Electron microscopic observation showed that there were more rough endoplasmic reticulum, mitochondria and lysosomes in Adv-hBMP-2 transfected MSCs compared to MSCs of other two groups. At the 3rd and 6th weeks after cell injection, ectopic bones were observed in muscles of nude mice of Group 1. Only fibrous tissue or a little bone was found in other two groups.

CONCLUSIONSBMP-2 gene transfected MSCs can differentiate into osteoblasts in vitro and induce bone formation in vivo.

Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; metabolism ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; genetics ; Cell Differentiation ; Genetic Therapy ; Goats ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Nude ; Osteogenesis ; physiology ; Staining and Labeling ; Tissue Engineering ; Transfection ; Transforming Growth Factor beta ; genetics

OBJECTIVE To study the methodology of achieving stable co-expression of drug-metab?olizing enzymes in the HepG2 cells by the piggyBac (PB) transposon system. METHODS N-terminal attachment of enhanced green fluorscent protein plasmid (pEGFP- N2) and 2A peptide linked recombinant PB transposon plasmid containing dual-genes encoding drug metabolizing enzymes cyto?chrome P450 3A4 (CYP3A4) and CYP2C19 (pPB-CYP3A4-2A-2C19) were transfected into HepG2 cells respectively by Lipofectamine?LTX reagent, GenJetTM (Ver.Ⅱ) reagent and Neon?Transfection System reagent, which were widely used for large-sized DNA fragments transfection. 48 h later, the transfection efficiency and cell toxicity were detected and compared between the three methods so as to find a method with relatively high efficiency and low toxicity for later transfection.Then,three groups of recombinant PB transposons-single-gene transposon (PB-CYP3A4), 2A peptide linked dual-gene transposon (PB-CYP3A4-2A-2C19) and multiple single-gene transposon mixture〔PB-CYP3A4, PB-CYP2C8, PB-CYP2A6, organic anion transporting polypeptide 1B1 PB transposon (PB-OATP1B1)〕-were transfected into HepG2 cells respectively with the above established method.The puromycin (Puro)-resistant and GFP positive cell clones were picked up and further cultured. The mRNA, protein and metabolic levels of drug-metabolizing enzymes in monoclonal cell lines were detected by quantitative real-time PCR,Western blotting and high performance liquid chromatography-tandem mass spectrometry respectively after screening by Puro and green fluorescence. Comparisons of different groups were made using statistical analysis. RESULTS The comparison of three different transfection methods indi?cated that the transfection efficiency of GenJetTMwas up to(94.2±2.5)% and (89.3±3.3)%,significantly higher than those of the other two methods (P<0.01), along with lower cytotoxicity. Then GenJetTMwas chosen for later transfection. In the Puro-resistant monoclonal cell lines of single transposon PB-CYP3A4,PB-CYP3A4-2A-2C19 groups,the mRNA,protein and enzyme activity levels of drug-metabo?lizing enzymes were significantly increased respectively.The recombinant transposon (PB-CYP3A4-2A-2C19) containing 2A peptide could achieve stable and efficient co-expression of two metabolizing enzymes CYP3A4 and CYP2C19,while the expression of drug-metabolizing enzymes remained unbal?anced and random in those of multiple single-gene transposon mixture group (PB-CYP3A4, PB-CYP2C8,PB-CYP2A6,PB-OATP 1B1)(CYP3A4 was expressed in some cell clones only).CONCLUSION GenJetTM could be an effective method for the PB recombinant transposon transfection into HepG2 cells, by which the PB transposon could mediate stable expression of drug-metabolizing enzymes. In terms of multi-gene expression,a low and unbalanced expression is found by multiple transposons co-transfection method,which is different from that by virus mediated method.In contrast,mono-PB trans?poson linked by 2A peptide can achieve stable expression of multi-genes.