OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.

METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.

RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.

CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers

Objective To determine the activities of 131I for treating differentiated thyroid carcinoma with diffuse pulmonary metastases ( DTC-DPM ) from the perspective of internal radiation dosimetry.Methods According to Medical Internal Radiation Dosimetry (MIRD) schema, the activity constraint,from which the whole bdy retention at 48 h should not exceed 2.96 GBq (2.96 GBq rule), was converted to dose-rate constraint(DRC) to lungs at 48 h ( DRCLU ·48 h ) in 131I therapy for DTC-DPM. Based on the assumption of DRCLU·48 h at 48 h in lung, the fractions of whole body activities ( F48 ), the effective half times of 131I in lungs ( TLL ) and the remainder of body ( TRB ) were 0.6-0.9, 20- 120 h, and 10- 20 h, respectively. The maximum safe activities of 131I for different human phantoms from the Organ Level Internal Dose Assessment (OLINDA) software were calculated. Results According to MIRD schema and 2.96 GBq rule, DRCLU ·48 h should not exceed 46.4 mGy/h in 131I therapy for DTC-DPM. Depending on varying F48 h,TLL and TRB, the maximum safe activities of 131I were 6.77-81.36, 5.29-56.20, 5.08-55.19 and 3.87-40. 52 GBq for the male adult, female adult, 15-year-old, and 10-year-old patients with DTC-DPM, respec tively. Conclusion Dosimetry-guided 131I therapy for DTC-DPM considers adequately the differences of 131I kinetics in individual patients and can adjust administered activities of 131I on the precondition of avoiding radiological pneumonitis and pulmonary fibrosis.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Imaging, Three-Dimensional ; Male ; Maxillofacial Injuries ; diagnostic imaging ; surgery ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Skull Fractures ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

OBJECTIVETo determine the inhibitory effect of 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human colon carcinoma cells and xenografts in nude mice, to observe its effect on CDH13 gene expression and methylation in the xenografts, and to explore the possible mechanisms.

METHODSHuman colon carcinoma cell line HCT116 cells were treated with 5-Aza-CdR, and the cell morphology was observe by phase contrast microscopy. The cell growth was assessed by MTT assay. A tumor-bearing mouse model was generated by subcutaneous inoculation of human colon carcinoma HCT116 cells into nude mice. The tumor growth in the nude mice was observed, the CDH13 gene expression and its methylation status in the tumors were detected using methylation specific PCR (MSP), RT-PCR, Western blotting and immunohistochemistry.

RESULTSAfter treatment with 5-Aza-CdR, the inhibition rate of the growth of cultured HCT116 cells was increased as the concentration was increasing. The growth of the xenografts in nude mice was significantly inhibited, and the methylated CDH13 gene was reactivated. After 4 weeks of 5-Aza-CdR treatment, no significant difference was found between the body weights of nude mice in the 5-Aza-CdR group [(18.06 ± 1.29) g] and control group [(17.07 ± 0.84) g], (P > 0.10), and the average volume of xenografts of the 5-Aza-CdR group was (907.00 ± 87.29) mm(3), significantly smaller than the (1370.93 ± 130.20) mm(3) in the control group (P < 0.005). No expression of CDH13 gene was found in the control group. The expression of CDH13 gene in the 5-Aza-CdR group was increased along with the increasing concentration of 5-Aza-CdR.

CONCLUSIONS5-Aza-CdR inhibits the growth of human colon cancer cells in culture and in nude mice, and induces the cancer cells to re-express CDH13 in nude mice. Its mechanism may be that demethylation of the methylated CDH13 promoter induced by 5-Aza-CdR restores CDH13 expression and thus inhibits the tumor growth in nude mice.

Animals ; Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cadherins ; genetics ; metabolism ; Cell Proliferation ; drug effects ; DNA Methylation ; HCT116 Cells ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; metabolism ; Tumor Burden ; drug effects

BACKGROUND AND OBJECTIVEColon cancer is one of the most common malignant tumors, and its pathogenesis is not fully understood. Transcriptional silencing by DNA methylation is believed to be an important mechanism of carcinogenesis. E-cadherin can suppress tumor cell invasion and metastasis, and is considered as an invasion/metastasis suppressor gene. Inactivation of E-cadherin gene often occurs in colon carcinoma. This study was to investigate the correlation between E-cadherin gene expression and the methylation status of E-cadherin 5' CpG islands in human colon carcinoma cell line HT-29, and to explore the mechanism of carcinogenesis of colon cancer.

METHODSImmunocytochemical dicho-step method and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression of E-cadherin protein and mRNA in HT-29 cells after 5-Aza-CdR treatment; methylation specific PCR was used to analyze the methylation status at promoter of E-cadherin gene.

RESULTSThe expression of E-cadherin gene could be restored by 5-Aza-CdR treatment, immunocytochemical staining showed the positive expression ratio of E-cadherin increased from (21+/-7)% (1 micromol/L) to (39+/-13)% (5 micromol/L); E-cadherin genes were methylated and not expressed in HT-29 cells in the colon carcinoma.

CONCLUSIONSE-cadherin methylation plays an important role in the carcinogenesis of colon carcinoma cells and can re-express after the treatment with 5-Aza-CdR.

Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cadherins ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; drug therapy ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; HT29 Cells ; metabolism ; pathology ; Humans ; RNA, Messenger ; metabolism

Objective To investigate the influence of high intensity interval exercise (HIIT) on peroxidation and vascular endothelial function for experimental hyperhomocysteinemia (HHcy) rats. Methods Thirty five male rats were randomly divided into 4 groups. Control group (n=8) was given ordinary feed. High methionine group (n=27) was given 3% methionine on this basis, and divided into model group, folic acid group and HIIT+ folic acid group, with 9 rats per group for 16 weeks. Serum homocysteine (Hcy) , content of plasma malondialdehyde (MDA) , hydroxyl radical (OH-), total antioxidant capacity (T-AOC), activity of glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) were measured, as well as the level of Nitric Oxide (NO), Nitric Oxide Synthase (NOS) and Endothelin 1 (ET-1) . The pathology of abdominal aortas was analyzed.Results Sixteen weeks after intervention, there was no significant difference between HIIT + folic acid group and the control group (P>0.05) . The levels of serum Hcy in the model group, folic acid group and the HIIT+folic acid group were (23.95±3.35) μmol/L,(8.73±0.60) μmol/L, and (6.19±0.34) μmol/L respectively (P<0.05) . Sixteen weeks after intervention, the content of MDA in HIIT+ folic acid group reduced, and there was no significant difference compared with the control group (P>0.05). The level of SOD and GSH-PX increased in HIIT+ folic acid group and folic acid group, and there was a significant difference compared with the model group. There were significant differences in activities of SOD and GSH-PX in HIIT+ folic acid group when compared with folic acid group (P<0.05). Compared with the control group, there were significant differences in levels of ET-1, NOS and NO in folic acid group (P<0.05), while there was no significant difference in the level of ET-1 and NOS between HIIT+folic acid group and control group (P>0.05) . Mild atherosclerotic lesions were observed in the HIIT+folic group. Conclusion High methionine diet can reduce the level of serum Hcy in HHcy rats, and high intensity interval exercise combined with folic acid intervention could reduce the level of serum Hcy, improve oxidative stress state, reduce the injury of endothelial function, and thus to alleviate atherosclerotic lesion.

Adolescent ; Adult ; Athletic Performance ; psychology ; Back ; physiology ; Electromyography ; Exercise ; physiology ; Humans ; Imagery (Psychotherapy) ; Male ; Young Adult

OBJECTIVETo observe the maintenance effect of polyglycosides of Tripterygium wilfordii (GTW) on remission in postoperative Crohn disease (CD).

METHODSFrom 2005 to 2007, 45 adult cases of postoperative Crohn disease were randomly divided into two groups, GTW group and mesalazine group, which received GTW and mesalazine treatment respectively. CD activity index (CDAI) and clinical markers were collected at 0, 3, 6, 12 months or at the onset of symptoms. Ileocolonoscopy was performed at the end of the trial (1 year after operation) or at the onset of symptoms, and recurrence score were recorded.

RESULTSNo clinical recurrence was ascertained in both groups at 3 months. Four patients (18.2%) in GTW group relapsed and 5 (21.7%) in mesalazine group relapsed at 6 months (P=0.530). Seven patients (31.8%) in GTW group and 9 (39.1%) in mesalazine group relapsed at one year (P=0.421). Ten patients (45.5%) in GTW group had endoscopic recurrence compared with 14 (60.9%) in mesalazine group at one year(P=0.231). There were no significant differences between two groups.

CONCLUSIONGTW is similar to mesalazine in maintenance of remission of postoperative Crohn disease.

Adolescent ; Adult ; Aged ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Crohn Disease ; drug therapy ; Glycosides ; therapeutic use ; Humans ; Male ; Mesalamine ; therapeutic use ; Middle Aged ; Phytotherapy ; Postoperative Period ; Recurrence ; Treatment Outcome ; Tripterygium ; chemistry ; Young Adult

OBJECTIVETo investigate the role of Rho-kinase signaling pathway in human airway smooth muscle cell (ASMCs) migration and cytoskeletal reorganization induced by endothelin-1 (ET-1).

METHODSPrimary cultured human ASMCs obtained by tracheal explant culture method were examined for cell migration in response to ET-1 treatment using modified Boyden chambers. The changes in actin cytoskeletal reorganization were observed under confocal laser scanning microscope, and the phosphorylation of myosin-phosphatase target 1 (p-MYPT1) was examined using Western blot analysis.

RESULTSAt the concentration of 0.1, 1, 10, and 100 nmol/L, ET-1 induced migration of the ASMCs, and 10 nmol/L ET-1 produced the most obvious effect (P<0.01). Rho-kinase inhibitor Y-27632 showed a dose-dependent inhibitory effect on ET-1-induced ASMC migration, and in cells exposed to 10 nmol/L ET-1, Y-27632 at 10 micromol/L significantly blocked ASMC migration (P<0.01). ET-1 (10 nmol/L) exposure resulted in reorganization of actin cytoskeleton and formation of stress fibers in the ASMCs, which were obviously inhibited by Y-27632. Compared with the control group, the AMSCs showed significant enhancement of p-MYPT1 protein expression after ET-1 exposure for 15 and 30 min (P<0.01), but prolonged exposure failed to result in the expression enhancement (P>0.05).

CONCLUSIONRho-kinase signaling pathway may play an important role in ET-1-induced ASMC migration and reorganization of actin cytoskeleton.

Amides ; pharmacology ; Bronchi ; cytology ; Cell Movement ; drug effects ; Cells, Cultured ; Cytoskeleton ; drug effects ; metabolism ; Endothelin-1 ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Humans ; Microscopy, Confocal ; Muscle, Smooth ; cytology ; Pyridines ; pharmacology ; Signal Transduction ; drug effects ; rho-Associated Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo evaluate the clinical value of iodine[131I] metuximab infusion combined with transcatheter arterial chemoembolization (TACE) for treating cases of post-intervention relapse of mid or advanced stage hepatocellular carcinoma (HCC).

METHODSSixty patients who were diagnosed between March 2009 and June 2010 with relapse of mid or advanced stage HCC following previous intervention with various standard clinical methods were recruited for study. The patients were randomly and equally divided into a control treatment group (CG; receiving TACE therapy alone) and an experimental treatment group (TG; receiving TACE combined with iodine [131I] metuximab injection). For all patients, licartin was first perfused into the tumor feeding artery and then the TACE procedure was performed 20 min later. Liver function markers and routine blood parameters, including alpha-fetoprotein (AFP) and clotting time, were examined at one week and one month after the treatment. Enhanced computed tomography or magnetic resonance imaging of the liver was performed at one month after treatment and thereafter on a bi-monthly follow-up schedule. The World Health Organization's tumor evaluation standard was used to assess the therapeutic effects in each group. Results of laboratory tests (pre- and post-treatment), reported complications, and side-effects were evaluated for their contributions to time of tumor progression (TTP) and survival time.

RESULTSPatients in the TG and CG groups had similar blood cell counts at pre-operative and 1-week postoperative time points. The TG group showed a significantly reduced level of AFP following treatment, but it was not significantly different from the level in the CG group. The TG group did however show significantly different levels of liver functional parameters (all P less than 0.05) and significantly higher TTP (4.84+/-4.11 vs. CG: 2.54+/-2.08 months; t = -2.13, P less than 0.05) and average survival time (7.05 vs. 5.15 months; x2 = 4.24, P = 0.039). The rates of partial response (PR), slight remission (MR), unchanged status (SD) and progressive disease (PD) were 16.7%, 37.5%, 25.0% and 20.8% in the TG group, and 8.7%, 17.4%, 21.7% and 52.2% in the CG group. The therapeutic effect rate (CR + PR + MR) and reaction rate (CR + PR + MR + SD) was significantly different between the two groups (P = 0.048). No serious adverse effects were reported.

CONCLUSIONTACE combined with iodine [131I] metuximab injection is a safe and effective procedure for prolonging the survival and TTP of patients with HCC relapse following prior therapeutic intervention.

Aged ; Antibodies, Monoclonal ; therapeutic use ; Carcinoma, Hepatocellular ; pathology ; therapy ; Chemoembolization, Therapeutic ; methods ; Female ; Humans ; Iodine Radioisotopes ; therapeutic use ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; therapy ; Treatment Outcome