Objective To study the antigenicity of human thyrotropin receptor(hTSHR)amino terminus (amino acid 29～280)and its association with Graves' disease.Methods Total thyroid RNA was prepared from human normal thyroid tissue.RNA was then reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and the recombinant plasmid was named pcDNA3.1/hTSHR188～940bp. Balb/c mice were immunized with peDNA3.1/hTSHR188～940bp. The levels of serum thyroxin,anti-TSHR antibody(TRAb)and thyroid stimulating antibody(TSAb)were measured,and the pathological changes of thyroid tissue were also observed.Results A 753 bp fragment encoding hTSHR ectodomain amino end was obtained after PCR amplification.Confirmed by Hind Ⅲ restriction enzyme digestion and DNA sequencing,pcDNA3.1/hTSHR188～940bphad been constructed successfully,with the correct sequence and direction of hTSHR188～940bp.In the Balb/c mice treated with pcDNA3.1/hTSHR188～940bp,elevated TRAb in week 6(0.148±0.018)were observed compared with those at week o(0.106±0.006,P＜0.01),and kept a higher level till week 10(0.134±0.011,P＜0.01).T4 and TSAb index values were significantly increased in week 10.Serum T4 concentration increased from(41.02±7.97)μg/L in week 0 to(62.20±12.77)μg/L in week 10(P＜0.01);TSAb index values rose from 0.864±0.076 at week 0 to 1.392±0.615(P＜0.01).Thyroid pathological examination showed that proliferated thyroid follicular epithelial cells and foll icular eapacity increased.Inflammatory cells were occasionally found.Conclusions There are antigen epitopes in hTSHR ectodomain amino acid 29～280,which can stimulate the production of TSAb.And the latter induces hyperthyroidism and Graves' disease like manifestations.It suggests that hTSHR ectodomain amino acid 29～280 is closely associated with Graves' disease,and maybe one of important etiological factors leading to the disease.

Objective Establish rabbit VX2 soft tissue tumor model,and treat it with percutaneous ethanol injection(PEI)under the guidance of magnetic resonance imaging.Make ready for the therapeutic evaluation with functional magnetic resonance imaging. Methods Fifteen healthy New Zealand white rabbits were included in this study.0.2 mL tumor tissue suspensions were injected into the rabbits’posterior limb.14 days later,all rabbits were underwent conventional MRI examination.PET were performed to all the tumors under the guidance of MRI in the next day of the examination.T2 WI was used as guidance and monitoring means.MR com-patible puncture needle with lateral hole was stabed into the lesion center,and inj ected anhydrous ethanol according to the volume of tumors’diameter (1 mL/cm )slowly.the tumors signal characteristics,morphological feature and pathological feature were ob-served pre and post-operation.Results All of the 1 5 rabbits were established soft tissue tumor model successfully;the success rate is 100%.The tumors were oval or round,3-4 cm in diam.MRI scanning showed low signal on T1 WI and high signal on T2 WI be-fore PEI.PEI was performed to all the tumors under the guidance of MR successfully with 3.5 mL ethanol injected into the tumors in average.T2 WI could monitor the ethanol in dispersion and distribution within the tumors clearly.Histologically,tumors were composed of large,uniform,oval/round cells arranged in solid nests which was intensive in the periphery of tumors.Necrosis tissue was apparent in the center of the tumors.10 days after operation,most tissue in the periphery of tumors was coagulative necrosis , only a few tumor cells left.Ranges of necrosis in the tumors center were obviously increased compared with pre-operation.Conclusion Rabbit VX2 tumor of soft tissue model is suitable for the therapeutic evaluation of tumor .It is an animal model which has the characteristic of simple to operate and high rate of suc-cessful.MR T2 WI can monitor the ethanol in dispersion and distribution within the tumors clearly.It is a good guidance and monitoring imaging method of tumor ablation.

OBJECTIVETo explore the effects of ytterbium citrate on human liver carcinoma HepG2 cell line and the potential mechanisms.

METHODSThe HepG2 cells were cultured with DMEM medium and divided into different groups in the following media, in serum-free medium as control, different concentration (0.01 - 5.00 mmol/L) [YbCit(2)](3-)+serum-free medium as treatment group, MTT assay was used to measure the viability of the cells; 2.00 mmol/L [YbCit(2)](3-)+serum-free medium was used as treatment group, and Hoechst 33258 staining was used to detect apoptosis in HepG2 cells. Differential proteomic analysis, assay of intracellular H(2)O(2) levels and mitochondrial transmembrane potential were performed to study the effects of [YbCit(2)](3-) on HepG2 cells and the potential mechanisms.

RESULTSThe data showed that 72 h treatment of [YbCit(2)](3-) at 2.00 - 5.00 mmol/L significantly inhibited cell proliferation, and the IC(50) was (2.46 ± 0.23) mmol/L. After treatment with 2.00 mmol/L [YbCit(2)](3-) for 48 h and 72 h, Hoechst 33258 staining demonstrated that [YbCit(2)](3-) induced significantly increased apoptosis in HepG2 cells. After treatment with 2.00 mmol/L [YbCit(2)](3-) for 72 h, two dimensional gel electrophoresis and MALDI-TOF mass spectrometry analysis revealed 14 differentially expressed proteins between [YbCit(2)](3-)-treated cells and the control cells. These proteins mainly included cofilin1, peroxiredoxin6, S100 calcium-binding protein A6, and proteasome 26S non-ATPase subunit 13 isoform 3 and so on. These proteins played important roles in the processes of anti-apoptosis, oxidation reduction, cell proliferation and protein degradation. The mitochondrial membrane potential were investigated, the results showed the red and green fluorescence ratio was 2.45 ± 0.28 in the control group, 1.56 ± 0.23 in 24 h group, 1.16 ± 0.18 in 48 h group, compared with the control, the differences were significant (F = 23.97, P = 0.001). The results of H(2)O(2) detection showed the fluorescence intensity was 20.00 ± 2.08 in the control group, 40.00 ± 5.50 in 24 h group, and 48.00 ± 2.03 in 48 h group, compared with the control, the differences were significant (F = 48.40, P = 0.000). The results indicated a significant reduction in mitochondrial transmembrane potential and significant increase in H2O2 generation were observed in [YbCit(2)](3-)-treated cells.

CONCLUSIONThese results suggested that [YbCit(2)](3-) could induce apoptosis of HepG2 cells through the mechanisms involving oxidative stress and mitochondrial dysfunction.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Oxidative Stress ; Proteome ; analysis ; Proteomics ; Ytterbium ; pharmacology

The objective of study was to explore the influence of high mobility group box 1 (HMGB1) on migration of cord blood CD34(+) cells and their mechanism of migration. The expressions of receptor for advanced glycation end products (RAGE), toll-like receptor-2 (TLR2) and TLR4 were detected by flow cytometry. The CD34(+) cells in umbilical cord blood (CB) were enriched by MiniMACS and were exposed to various concentration of HMGB1 (10, 50, 100, 1, 000 ng/ml), then the migration effect of HMGB1 on umbilical cord blood (UCB) CD34(+) cell count was determined by microscopy, the chemotactic index was calculated. The CD34(+) cells untreated with HMGB1 were used as control. The results indicated that the purity of the isolated CD34(+) cells was more than 98%. The HMGB1 could promote the migration of CD34(+) cells, and the migration effect of HMGB1 on CD34(+) cells in certain concentrations gradually increased along with raise of concentration, the strongest effect was observed in concentration of 100 ng/ml, there was significant difference as compared with control (p < 0.01). Anti-RAGE antibody partially inhibited the migration effect of HMGB1 on CD34(+) cells. It is concluded that the HMGB1 in certain concentration can enhance migration of CD34(+) cells, which may be mediated through RAGE.
Antigens, CD34 ; Cell Movement ; drug effects ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; drug effects ; HMGB1 Protein ; pharmacology ; Humans ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Signal Transduction

OBJECTIVETo identify the distribution and differentiation of ABL kinase domain mutation in the Chinese Han nationality imatinib resistant chronic myeloid leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL).

METHODSBone marrow or peripheral blood samples of 112 imatinib resistant CML patients and 21 Ph(+)ALL patients were obtained from the first affiliated hospital of Soochow university according to local law. Total RNA was extracted from the mononuclear cells using a TRIzol reagent. ABL kinase domain (KD) mutation was detected by direct sequencing.

RESULTSOf the 112 imatinib resistant CML patients, 54.46%(61 cases) had ABL KD mutation. Twenty-three mutants were identified in 20 amino acid sites and 23.21% (26 cases) ABL KD mutations were in P-loop region. ABL KD mutations were also detected in 71.43% (15 cases) imatinib resistant Ph(+)ALL patients, with 10 mutations in 8 amino acid sites. The most frequent mutation was T315I (28.57%), followed by E255K/V (19.05%) and Y253F/H (14.29%). The frequency of T315I was much higher in imatinib resistant Ph(+) ALL than that in imatinib resistant CML (P = 0.001). Ph(+)ALL with additional chromosomal aberrations also had a higher rate of ABL KD mutation than that of CML (P = 0.010). Ph(+)ALL gained ABL KD mutation faster than CML (P < 0.010).

CONCLUSIONChinese imatinib resistant CML and Ph(+)ALL patients had different characteristics in ABL KD mutation. The rate of ABL KD mutation in Ph(+)ALL with additional chromosomal aberrations was much higher than that of CML with additional chromosomal aberrations.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Benzamides ; pharmacology ; Chromosome Aberrations ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Middle Aged ; Mutation ; Philadelphia Chromosome ; Piperazines ; pharmacology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Protein-Tyrosine Kinases ; genetics ; Proto-Oncogene Proteins c-abl ; genetics ; Pyrimidines ; pharmacology ; Young Adult

OBJECTIVETo apply echocardiography in diagnosis of isolated noncompaction of ventricular myocardium (INVM) in adults.

METHODSSix patients with INVM underwent echocardiographic examination using HP5500 ultrasound system with the frequency of 2 approximately equals 4 MHz, and the observation was focused on ventricularmyocardium and endocardium at one-third of the apex.

RESULTSAll 6 patients showed typical echocardiographic images characterized by an altered structure of the ventricular myocardium with extremely thickened, hypokinetic segments consisting of two layers: a thin, compacted epicardium and an extremely thickened endocardium, resulting in an extremely thickened ventricular wall.

CONCLUSIONEchocardiography should be the first choice to be applied in diagnosis of INVM, which is a distinct entity of specific cardiomyopathy in adults.

Adult ; Cardiomyopathies ; congenital ; diagnostic imaging ; pathology ; Echocardiography, Doppler, Color ; Female ; Heart Ventricles ; diagnostic imaging ; pathology ; Humans ; Male ; Middle Aged ; Myocardium ; pathology

Adolescent ; Adult ; Constriction, Pathologic ; diagnostic imaging ; surgery ; Female ; Humans ; Male ; Peripheral Vascular Diseases ; diagnostic imaging ; surgery ; Renal Veins ; diagnostic imaging ; pathology ; Stents ; Ultrasonography

OBJECTIVETo study the methylation status of fragile histidine triad (FHIT) gene promoter in patients with myelodysplastic syndrome (MDS) and its clinical relevance.

METHODSMethylation-specific PCR (MSP) was used to detect FHIT promoter methylation in bone marrow samples from 54 MDS cases.

RESULTSHypermethylation of FHIT promoter was detected in 26 cases (48.1%). Association was not found between FHIT gene hypermethylation and sex, hematologic parameters and chromosomal abnormalities of MDS patients, but found between FHIT gene hypermethylation and age of the MDS cases. Although significant difference was not observed in the frequencies of FHIT gene hypermethylation among patients with refractory anemia/refractory anemia with ringed sideroblasts (RA/RAS) (1/6, 16.7%), refractory anemia/refractory anemia with ringed sideroblasts (RCMD) and refractory cytopenia with multilineage dysplasia with ringed blasts (RCMD-RS) (6/19, 31.6%), refractory anemia with excess blasts-1 (RAEB-1) (7/11, 63.6%), refractory anemia with excess blasts-2 (RAEB-2) (4/7, 57.1%) and refractory anemia with excess blasts in transformation/acute myeloid leukemia (RAEBt/AML) (8/11, 72.7%)(chi-square=8.417, P=0.077), it was observed in patients in early stages (RA/RAS and RCMD) (7/25, 28.0%), advanced stages (RAEB-1 and RAEB-2)(11/18, 61.1%) and RAEBt/AML (8/11, 72.7%) (chi-square=7.938, P=0.019). Furthermore, there was a positive correlation between the frequency of FHIT gene hypermethylation and different IPSS groups (chi-square=10.110, P=0.018).

CONCLUSIONFHIT gene hypermethylation might be one of the molecular events involved in the disease progression of MDS.

Acid Anhydride Hydrolases ; genetics ; Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Base Sequence ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Myelodysplastic Syndromes ; classification ; genetics ; pathology ; Neoplasm Proteins ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics

The study was aimed to explore the influence of co-culture ex vivo of CD34+ cells from two units of cord blood (CB) on the homing-related adherent molecule expression of each other. Mesenchymal stem cells (MSCs) were obtained from human bone marrow. Two units of CB CD34+ cells were co-cultured on 12 Gy gamma-ray irradiated MSC layer. Their adherent molecule expressions were assessed by flow cytometry. The results showed that the purity of the isolated CD34+ cells was (98.25+/-0.93)%. After co-culture on MSC layer for 6 days, the proportion of CD34+ cells of each unit was dropped to (60.4+/-6.32)% and (60.2+/-5.12)% respectively, but there was no significant difference from the control groups. The expressions of CD44, CD62L, CD184 and CD26 on CD34+ cells of each unit remained unaffected. The expression of CD162 was downregulated and CD54 was first increased but then dropped to the level before co-culture. But there was no significant difference between the experimental and control groups. In conclusion, co-culture of CD34+ cells from two units of CB may have no effects on the adherent molecule expressions of each other.
Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; Cell Adhesion Molecules ; metabolism ; Coculture Techniques ; Fetal Blood ; cytology ; metabolism ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology

This study was aimed to investigate the therapeutic efficiency and complications after allo-hematopoietic stem cell transplantation (allo-HSCT) with reduced-intensity conditioning regimens in hematologic malignancies. 10 patients (6 CML patients, 2 AML patients, 1 ALL patient and 1 NHL patient) underwent related allogeneic hematopoietic stem cell transplantation with reduced-intensity conditioning regimens. The conditioning regimens consisted of "FLU + CY + TBI" basically and was appropriately improved in accordance with status of patients. Cyclosporin A (CsA) and mycophenolate mofetil (MMF) were used to prevent the graft-versus-host disease (GVHD). Detection of bone marrow cells, chromosomes, fused gene, ABO blood group and STR-PCR were used to observe engraftment, relapse, GVHD, transplantation- related complications (TRC) after transplantation and to evaluate patients quality of life. The results showed that the 10 patients successfully accepted the transplantation and their primary diseases were cured. In one patient, severe pulmonary infection happened, and in another one CMV infection occurred. Grade IV of acute GVHD occurred in one case and grade I of acute GVHD in 2 cases, the no chronic GVHD appeared. 5 patients relapsed after transplantation at various time points, the donor lymphocytes infusion (DLI) or drugs rescued these 5 patients. During median follow-up of 5 - 35 months, 2 out of which died, 8 survived, the overall survival rate was 80%, and the survivors live in a high-quality life. In conclusion, the hematopoietic stem cell transplantation with reduced intensity conditioning regimens was feasible with relatively low toxicity for recipients. GVHD and TRC were low, and life quality of patients after transplantation was high. DLI could cure the primary diseases even relapsed after transplantation.
Adult ; Cyclosporine ; administration & dosage ; Female ; Graft vs Host Disease ; prevention & control ; Hematologic Neoplasms ; therapy ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Lymphocyte Transfusion ; Male ; Middle Aged ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; Transplantation Conditioning ; methods