Objective To investigate the telomerase activity and abnormality of p16 gene in liver metastatic tumors of colorectal carcinoma. Methods Telomerase activity was detected by a non isotopic PCR based TRAP assay (Telomeric repeat amplification protocol) and the homozygous deletions of p16 gene was detected by a semiquantitative multiple PCR in tissue samples from 24 liver metastases of colorectal carcinoma. Results Telomerase activity were observed in 19 (79 2%) out of the 24 colorectal carcinoma patients. There was no correlation hetween the telomerase activity, the number of metastasis, differentiation, liver fibrosis and HBsAg status of the patients. Homozygous deletions of p16 gene were found in 9 out of 24 (37 5%) patients, and homozygous deletions of p16 gene was significantly correlated with telomerase activity.Conclusion The abnormality in telomerase activity and homozygous deletions of p16 gene may be important for elucidating in liver metastases of colorectal carcinoma.

Objective To study the direct effect of parathyroid hormone(PTH) on osteoclasts (OCs) differentiation and bone resorption in vitro and to explore the relationship between PTH and expression of RANKL (ligand of receptor activator of nuclear factor kappa B)gene and OPG (osteoprotegerin) gene in osteoblasts (OBs) in order to provide some theoretic evidence for understanding the pathogenesis of high-transferred renal osteodyslrophy (ROD). Methods PTH was directly used to induce OCs differentiation from whole bone marrow of C3h mouse by using OCs formation assay and then inspected the bone resorption of these OCs in vitro by using pits assay. Multiple RT-PCR assay was used to examine the expression of RANKL-mRNA and OPG-mRNA in different PTH concentrations and different treating times by PTH. Results (1) PTH in vitro could induce OCs formation form whole bone marrow of wild-type C3h mice.In a certain concentration scope, as the concentration of PTH increased, the number of differentiated OCs and the degree of bone destruction of OCs increased. (2) Expression of RANKL-mRNA and OPG-mRNA was in dose-dependent and time-dependent manner within a certain range of concentration and time. Conclusion PTH in vitro mediates OCs differentiation and bone resorption probably through the up-regulated expression of RANKL-mRNA and OPG-mRNA.

Objective To explore the clinical characteristics of adrenoleukodystrophy (ALD). Methods The clinical date of 14 ALD patients were analyzed retrospectively. Results The 14 ALD patients were boys,the symptoms occurred slowly from 1 to 13 years old. The clinical manifestations of all the ALD patients were mainly mental retardation and disturbances of movement of extremities,9 cases had visual disorders,6 cases had auditory disorders and 13 cases had dysarthria,5 cases had seizure and 6 cases had increased skin pigmentation. The level of plasma very long chain fatty acids (VLCFA) in 5 csaes were increased at some degree.The brain CT from 2 cases and MRI from 12 cases showed butterfly-like focus in periventricle areas in the white matter bilaterally. Lace-like high intensity lesions were observed in 3 cases. Conclusion The clinical features of ALD are progressive dysnoesia,limbs dyskinesia,hypropsia,hearing loss etc,dysfunction of adrenal cortex,the level of plasma VLCFA increased,and with characteristic changes of skull imageological.

0.05),but both groups reached a better result when compared with the auto-HSCT group(P

AIM: To investigate the possibility of simultaneously ex vivo generating cytomegalovirus (CMV) pp65 and Epstein - Barr virus (EBV) - specific cytotoxic T lymphocytes (CTL) from human umbilical cord blood (CB). METHODS: Mononuclear cell derived from CB (CBMC) was used to construct EBV - transformed B-lym- phoblastoid cell lines (BLCL). Then BLCL were transduced with a recombinant retrovirus encoding pp65, the immunodominant CMV polypeptide. CBMC from the same CB donor were stimulated with pp65 - expressing BLCL (BLCLpp65) weekly for 5 - 6 weeks. Chromium release assays (CRA) were performed to detect the specific cytotoxicity of the CTL against EBV and CMV. RESULTS: Western blot analysis and immunocytochemistry confirmed that BLCLpp65 could simultaneously express CMVpp65 and EBV antigen. CRA results showed that the generated CTL possessed specific cytotoxic against EBV and CMV, and the cytotoxicity was mediated by CD8+ CTL. CONCLUSION: BLCLpp65 can be used as antigen - presenting cells to stimulate expansion of EBV and CMV specific CTL simultaneously from the predominantly native T cell population in CB.

Objective To explore the influencing factors and possible mechanism of osteonecrosis of the femoral head(ONFH)in patients with leukemia after allogeneic hematopoietic stem cell transplantation (Allo-HSCT).Methods One hundred and two patients with leukemia who received Allo-HSCT between January 2003 and October 2007 were evaluated for ONFH and graft-versus-host disease(GvHD)within 2years after transplantation,and the dosage of methylprednisolone(MP)in every patient from the first diagnosis of leukemia to 2 years after Allo-HSCT was calculated.For patients who suffered acute GvHD(aGvHD) and chronic GvHD(cGvHD),the serum leveh of soluble ligand of receptor activator of nuclear factor kappa B (sRANKL)which was index for differentiation of osteoclast(OC),tartrate-resistant acid phosphatase-5b(TRAP-5b)and C-terminal telopeptide of collagen Ⅰ(CTP-Ⅰ)which both were indexes for metabolism of OC were detected by enzyme-linked immunosorbent assay in different stages of MP dosage.According to these results.possible factors for ONFH after Allo-HSCT were analyzed.Results Seven in 102 patients after Allo-HSCT had experienced ONFH within 2 years,the incidence was 6.9%(7/102),which was not related to age,gender,types of leukemia and donor.ONFH mainly developed in patients with aGvHD and cGvHD,and the incidence could be achieved to 21.9%(7/32)which Was much higher than that in patients with no GvHD,merely aGvHD or merely cGvHD(P<0.05).In patients with aGvHD and cGvHD,the average dosage of MP in ONFH(+)was(232.7±28.6)mg/kg which was extremely higher than that in ONFH(-)(115.1±16.9)mg/kg(P<0.05).The serum levels of sRANKL,TRAP-5b and CTP-Ⅰ were also higher when ONFH happened than ahead of pretreatment and 28 days after transplantation(-)(P<0.05),and they were closely related to the dosage of MP(correlation coefficient was 0.597,0.664 and 0.682 respectively,P<0.05).Conclusions After Allo-HSCT,ONFH is related to the dosage of MP in treatment of GvHD.MP may be responsible for enhancement of OC differentiation and metabolism in leukemia patients,and ultimately induced the onset of ONFH.

AIM: To determine the expression of membrane-bound B lymphocyte stimulator((BLyS)) and its mRNA in peripheral blood mononuclear cells(PBMCs) from individuals with systemic lupus erythematosus(SLE),and to investigate the effect of dexamethasone on(BLyS) expression.METHODS: PBMCs were obtained from 25 individuals with SLE(mean age of 31.40?14.23) and 20 female healthy volunteers(mean age of 28.20?10.36).They were randomized into dexamethasone((1 ?mol/L)) group and media group.PBMCs were gathered at 0,6,12 and 24 h for(BLyS) mRNA assessment using reverse transcription-PCR(RT-PCR).PBMCs were also collected at 72 h for membrane-bound(BLyS) protein detection using flow cytometry(FACS) and direct immunofluorescence.RESULTS:(1) The expression of(BLyS) mRNA and membrane-bound protein were significantly higher in PBMCs from individuals with SLE than that in PBMCs from healthy controls(0.40?0.18 vs 0.27?0.20,P

Objective To explore the clinical features of infection in multiple myeloma (MM)undergoing autologous hematopoietic stem cell transplantation (ASCT). Methods Thirty-seven patients with MM undergoing ASCT were retrospectively analyzed for type and time of infection, pathogen, and outcome. Results Fifty-nine cases of infectious complications occurred in 33 patients (89. 2% ) after ASCT, with 34 cases (57.6%) of bacterial infections in 30 patients, 15 cases (25.4%) of fungal infections in 12 patients, 4 cases (6. 8% ) of cytomegalovirus (CMV) infection, 3 cases (5. 1% ) of herpes zoster virus infection and 3 cases (5. 1% ) of HBV reactivation. The proportion of bacterial infection, fungal infection and virus infection were 62. 8%, 28.6% and 8. 6% respectively in the early stage after ASCT, and 50. 0%, 20. 8% and 29. 3% respectively in the median stage. Response to first-line antibiotic therapy was seen in 38 cases (64. 4% ). Infection-related mortality was 8. 1% (3 cases). Conclusions The incidence of infection in MM patients undergoing ASCT is high and they are susceptible to all pathogens. It is important to choose the right antifungal agents as quickly as possible to reduce infection-related mortality.

Objective To investigate the efficacy and safety of bortezomib-based induction regimen followed by autologous hematopoietic stem cell transplantation (ASCT) in pationts with multiple myeloma (MM). Methods A retrospective analysis was performed upon clinical data of 62 MM patients who received bortezomib-based induction regimen followed by ASCT from June 2006 to June 2011.All patients were followed up to September 30,2011.Results Overall response rate [ complete remission (CR) + near complete remission (nCR) + partial remission (PR) ],≥ nCR rate (CR/nCR) and CR rate of postinduction with bortezomib-based regimen were 88.7％,66.1％ and 24.2％,respectively.After ASCT,CR rate and CR/nCR rate were increased to 50.0％ and 82.3％,respectively,with significant differences (P =0.003 and P =0.032).The median time of neutrophil and platelet engraftment was 12.0 (9-43) days and 13.5 (0-120) days,respectively. Significances were found in neutrophil and platelet engraftment between MM patients with and without prior exposure to alkylating agents. Furthermore,engraftment of neutrophil and platelet in patients receiving peripheral blood stem cell transplantation were faster than those receiving bone marrow transplantation.No unexpected side effects occurred.The median time of follow-up was 26.5 (7-61) months.The median overall survival (OS) was not reached and the median progression-free survival (PFS) was 30 months.There were significant differences in OS and PFS between patients obtaining CR/nCR and those with ≤ PR before ASCT.Conclusions Bortezomib-based induction regimen can improve the efficacy of ASCT in MM patients.The side effects are tolerant.Higher response quality before ASCT can translate to high rates of OS and PFS following high-dose therapy and stem cell transplantation.

Objective To investigate the effect of bortezomib on the proliferation and apoptosis in leukemia cell line NB_4-R2 in vitro and provide some new evidences for the treatment of acute promyelocytic leukemia APL with ATRA-resistant using bortezomib. Methods NB_4-R2 cells were incubated with bortezomib at different does for 48 h. The proliferation capacity was measured by MTT assay, the morphology of cell apoptosis observed with Hoechst33342 staining by fluorescence microscopy and the percentage of apoptosis calculated by flow cytometry. The expression of apoptosis protein of cleaved (poly ADP-ribose polymerase, PARP) and Caspase-3 were determined by Western blotting. Results The proliferation of NB_4-R2 cells were obviously inhibited by bortezomib in vivo and the role of inhibition was a does-dependant manner within the scope of the bortezomib concentration from 1-5 μg /L.The incidence of inhibition was up to 74.9 % at the bortezomib concentration of 5 μg/L. Within this scope of the bortezomib concentration mentioned above, the role of inhibition of proliferation of NB_4-R2 cells mainly showed an increase of the late apoptosis, and the percentage of apoptosis was up to 78.9 %. In the meaning time, the expressions of the apoptotic protein of cleaved PARP and Caspase-3 were up-regulated in NB_4-R2 cells after treated with bortezomib by Western blotting assay. Conclusion Bortezomib can inhibit the proliferation of NB_4-R2 cells in vivo by inducing cell apoptosis.