Objective To investigate methylation of the P15INK4B and p27kipl genes in human mantle cell lymphoma cell line Mino,to evaluate the effects of decitabine on demethylation of p15INK4B and p27kip1 genes and on apoptosis of Mino cells and its relative mechanism. Methods Mino cells were after treated with various concentration of decitabine, the cell viability, cell cycle distribution or the apoptosis of Mino cells was respectively analyzed by trypan dye-exclusion assay or flow cytometry.The mRNA and protein expression of p15INK4B 、p27kip1 and bcl-2 were studied by RT-PCR or Western blot, respectively.Methylation of the p15INK4B and p27kip1 genes in Mino cells were determined by PCR using the methylation specific primer(MSP).Results Decitabine significantly inhibit the cell growth,induced G1 arrest and promoted apoptosis of Mino cells. The expression of p15INK4B and p27kipl mRNA were both significantly increased,wheres bcl-2 mRNA was decreased, After treatment with 6.4,3.2,1.6 mmol/L decitabine for 72 h,the methylationg of p15INK4B gene were decreased to 25.2 ％ 、48.2 ％ and 65.0 ％respectively,in the meantime,the methylationg of p27kip1 gene was decreased to 20.21％ 、50.2 ％ and 70.0 ％. Conclusion The hp15INK4B and p27kip1 genes of Mino cells are methylated and down-regulated.Decitabine can inhibit the proliferation and induce the apoptosis of Mino cell lines,with the reduction of bcl-2 gene and demethylation of p15INK4B and p27kip1 genes.

Objective To establish the model of pancreatoduodenal allotransplantation in pigs with enteric drainage (ED) and portal venous drainage (PVD). Methods Forty-six hybrid landraces were divided into two groups (donor and recipient groups) randomly, for pancreatoduodenal allotransplantation. Donors were perfused via abdomial aorta without clamping the portal venous outflow with UW solution after heparinization. Whole pancreatoduodenal graft was harvested with segments of abdomial aorta and portal vein and shaped under cold UW solution. Then, the end-to-end anastomosis was performed with the donor iliac artery bifurcation “Y” graft to the recipient superior mesenteric arteries and celiac artery. Furthermore, type Ⅰdiabete model was made by removal of the recipient pancreas. The venous anastomosis was reconstructed between the donor portal vein and the recipient superior mesenteric vein. Meanwhile, the end-to-side anastomosis was performed with the donor common iliac artery bifurcation “Y” graft to the recipient abdomial aorta and the side-to-side intestinal anastomosis was performed between the donor duodenum and the recipient jejunum. External jugular vein was intubated for transfusion. The levels of blood glucose, insulin and glucagon in blood were measured before and during the operation and 1, 3, 5, 7 d after operation. Results Twenty-three cases of pancreatoduodenal allotranplantations were performed on pigs. One died from complication of anesthesia. Success rate of operation was 95.7%.Complications of operation happened in 2 cases in which one was phlebothrombosis, incidence 4.5% and the other was duodenojejunal anastomotic leak, incidence 4.5%. The level of blood glucose increased within 30 min and recovered on the 2nd day after removal of pancreas. The levels of insulin and glucagon decreased within 30 min and recovered on the 2nd day after removal of pancreas. Rejection curred at the 1st day and reached the worst level on the 9th day after transplantation without the change of insulin and glucagon in blood and clinical symptoms of rejection. Conclusion Pancreatoduodenal transplantation in pigs can treat type Ⅰ diabete. ED and PVD can keep the function of endocrine in normal. The techni- que of duodenal transplantation with ED and PVD may pave the way for the further development of pancreas transplantation in clinic.

· AIM: To investigate the expression and the possible implication of CD40/CD40L costimulatory molecules in erythema nodosum of patients with Beh(c)et's disease.· METHODS: Sampling was done from erythema nodosum of 5 patients with Beh(c)et's disease and normal skin of 2 healthy individuals. Immunohistochemical staining was performed to examine the expression of CD4, CD8, CD19, CD68, HLA-DR,CD40 and CD40L molecules in the obtained tissues.· RESULTS: Approximately 90% of epidermic cells in erythema nodosum expressed CD40 molecule. In the dermis and subcutaneous tissue, a significantly increased number of CD4+Tcells, CD8+Tcells, CD19+cells, CD68+cells, HLA-DR+cells,CD40L+cells, and CD40+cells were observed in the erythema nodosum as compared with that in normal skin. Double staining showed that CD40L molecules were expressed on 45% of CD4+T cells. CD40 molecules were expressed on 100% CD68+ cells and 59.2% of HLA-DR+cells respectively.· CONCLUSION: A number of CD40/CD40L costimulatory molecules are upreguiated in the erythema nodosum of patients with Behcet's disease.

Abdominal Pain/etiology* ; Acute Disease ; Appendicitis/diagnosis* ; Diabetes Mellitus ; Diabetic Ketoacidosis/diagnosis* ; Diagnostic Errors/adverse effects* ; Humans ; Insulin

Objective To summarize radiographic manifestations of granulocytic sarcoma (GS),in order to investigate its radio-graphic feature.Methods Nine patients of GS confirmed by surgery pathology and immunohistochemistry were collected.There were four cases underwent CT scan,2 underwent MRI,3 underwent CT and MRI meanwhile.Results There were 4 cases in the lymph nodes,4 cases in the bone,1 case in the enterocoelia.The density of enlarged lymph node was inhomogeneous,and smaller lymph node was homogeneous density,both enhanced obviously.The density or intensity of the lesions occurred in the canalis verteb-ralis and paravertebra was also homogeneous,slightly enhanced,with bone and medulla ossea invasion.Lesions in the enterocoelia was inhomogeneous desity with clear boundary,enhanced inhomogeneously.Conclusion GS is a rare extramedullary malignant tumor composed of immature myeloid cells.CT and MRI can provide a certain diagnostic value.The perplexity in diagnosing such lesions highlighted the need of careful interpretation of all clinacal,radiographic,histopathological and immunochistochemical details as it is one of the most frequently misdiagnosed disorder.

Objective To choose proper proton magnetic resonance spectroscopy(~1XH-MRS) parameters to fit for practical femoral marrow cavity and to produce short-timed,well-repeated and excellent ~1H-MRS images.Methods The tentative study of ~1H-MRS on the normal femoral bone marrow in 26 volunteers was performed with a 1.5 T MR after the informed consent.The single-voxel spectroscopy and stimulated echo acquisition mode were used for ~1H-MRS collection.~1H-MRS parameters for 12 volunteers were 128 acquisitions,1 cm?1 cm?1 cm volume of interest(VOI)size and repeatedly 2—3 times within the same location.~1H-MRS parameters for another:14 volunteers were different numbers of acquisition (128 and 256 times,respectively)and different VOI sizes(2 cm?2 cm?2 cm and 1 cm?1 cm?1 cm, respectively).Results For ~1H-MRS with 1 cm?1 cm?1 cm size of VOI and 128 times of acquisition with the full width haft max of water≤8—12 Hz,the base-line was steady and the signal-noise ratio was high up to 11.31.~1H-MRS was different in the different femoral locations showing the maximum peak sites at near 0.90 ppm(?10~(-6))or 1.65 ppm,but~1H-MRS within the same location was always same or similar with different VOI sizes(1 cm?1 cm?1 cm or 2 cm?2 cm?2 cm)or different numbers of acquisition(128 or 256 times).~1H-MRS acquisition time was not related with the size of VOI but with the numbers of acquisition.128 and 256 times of acquisition cost 199 s and 391 s,respectively.Conclusion With the technique of small size of VOI(1 cm?1 cm?1 cm)and decreased numbers of acquisition(128 times),it is propable to get well-repeated and excellent ~1H-MRS within less time.It is also more practical for clinics to achieve ~1H-MRS of the femoral marrow with the proper technique.

Seven compounds were isolated from fermentation extract of cave-derived Metarhizium anisopliae NHC-M3-2 by silica gel, semi-preparative HPLC and other chromatographic methods. Their structures were elucidated by UV, IR, MS and NMR methods as 2,3-dehydroindigotide G (1), (-)-regiolone (2), naphtho-γ-pyrone (3), indigotide G (4), indigotide B (5) destruxin A (6) and destruxin B (7). Compound 1 is a new glycoside naphthopyranone compound. The anti-hepatitis B virus (HBV) activity of these compounds was evaluated. The EC₅₀ and CC₅₀ of compound 3 against HBV were 4.5 μmol·L^-1 and 92.3 μmol·L^-1, respectively. This is the first report of the antiviral activity of compound 3.

Adult ; Aged ; Animals ; Female ; Humans ; Kidney Calculi ; surgery ; Male ; Middle Aged ; Nephrostomy, Percutaneous ; instrumentation ; methods ; Pyonephrosis ; surgery ; Swine ; Swine, Miniature ; Ultrasonography, Interventional

OBJECTIVETo construct E1-deletion and replication-defective human type 5 recombinant adenovirus vector and to study the effect of p16INK4a on proliferation and aging of A549 cells.

METHODSp16INK4a cDNA was cloned into pAdCMV to construct recombinant pAdCMV p16INK4a, which was co-transfected into 293 cell together with pJM17. The recombinant p16INK4a adenovirus (Ad-p16INK4a) was generated by homologous recombination and identified with duplex PCR. Lung cancer cell A549, which has a homozygous deletion of p16INK4a gene, was infected with the prepared Ad-p16INK4a virus. X-gal staining and TRAP-ELISA were used for detecting senescence-associated beta-galactosidase and telomerase activities in A549 cells.

RESULTSImmunohistochemical staining and Western blot indicated that p16INK4a gene was transferred into A549 cell with more than 95% efficiency by recombinant adenovirus and p16INK4a protein was expressed at a high level- p16INK4a could markedly inhibit growth of A549 cells, induced expression of senescence-associated beta-galactosidase and suppressed telomerase activity in A549 cells.

CONCLUSIONRecombinant adenovirus vector could efficiently mediate transfer and expression of foreign genes in human cell and could be used for gene immunization and gene therapy; p16INK4a could inhibit A549 cell growth and induce its replicative senescence.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Cellular Senescence ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; physiology ; Gene Expression ; Genetic Vectors ; Humans ; Recombination, Genetic ; Transfection

OBJECTIVETo review the preliminary clinical experience with high-field-strength intra-operative magnetic resonance imaging (iMRI) in the endoscopic chordoma operation with transsphenoidal or transoral approach.

METHODSFrom January 2009 to December 2010, 23 patients [range, 29 - 64 years, mean age (42 ± 3) years] of chordoma were operated with endoscopic transsphenoidal or transoral approach and examined intraoperatively with a movable 1.5 T iMRI magnet. Tumor size range was 2.0 - 5.7 cm, mean (3.5 ± 0.8) cm. A navigation system based on iMRI was used in 20 cases.

RESULTSiMRI scan were performed in each operation from 1 time to 5 times. Neuronavigation system were used in 20 operations and the data renewed in 12 cases by the information from iMRI. In 15 of 23 patients, iMRI had revealed residual lesions and resulted in 12 cases further treatment, eventually, 9 tumors were totally removed and 3 tumors were further removed. The ratio of total removal tumor was enhanced to 73.9% (17/23) from 34.8% (8/23). Among 15 cases of partial chordoma removal detected by scanning in operation, 9 were huge chordoma. The residual of huge chordoma detected by scanning in operation was 9/11, and other chordoma contributed to 6/12. There were no iMRI related safety issue or accident recorded in this study.

CONCLUSIONSHigh-field-strength iMRI provide high-quality images of tumor resection that allows intraoperative modification of the surgical strategy. Combined with the navigation system, iMRI is helpful to maximize the resection of the chordoma and benefit for the safety of endoscopic operation.

Adult ; Chordoma ; surgery ; Endoscopy ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Neuronavigation ; methods ; Pituitary Neoplasms ; surgery ; Sphenoid Sinus ; surgery