Objective To summarize the diagnostic and therapeutic experience of a patient with chronic graft versus host disease (cGVHD) related polymyositis (PM) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods A patient with acute lymphocytic leukemia in com- plete remission received sibling allo-HSCT,and cyclosporine and methotrexate were adopted to pre- vent GVHD.Results Eleven days after HSCT,WBC＞0.5?10~9/L,13 days after HSCT,PLT＞20?10~9/L;27 days after HSCT,chromosome analysis of bone marrow cells showed 99% donor type. Seventeen days after HSCT,Ⅰ~0 acute GVHD of skin occurred,and it was cured by intravenous injec- tion of dexamethasone and methotrexate.Eight months after HSCT,cGVHD of liver happened.Al- though treated by tacrolimus and azathioprine,enzymes of liver were still elevated.At last,tacrolimus combined with sirolimus were used,and enzymes of liver subsided gradually.However,the serum creatine phosphokinase (CK) began to rise from 9 U/L to 3010 U/L,and fatigue all over the patient occurred.Finally,the symptom relapsed,and disability involved with the origin of limbs appeared. The electromyogram and magnetic resonance imaging of concerned muscles confirmed the PM diagno- sis.Although treated with methylprednisolone and plasma exchange,the patient died due to asphyxia, while the CK as high as 21 010 U/L.Conclusion PM is a rare kind of manifestations of cGVHD. When the key muscle tissue was involved,the prognosis is poor.

The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.
Amino Acid Sequence ; Chromatography, High Pressure Liquid ; methods ; Interferons ; standards ; Mass Spectrometry ; methods ; Molecular Weight ; Oxidation-Reduction ; Peptide Mapping ; Protein Processing, Post-Translational ; Reference Standards

Objective To understand the prevalence of smoking in different sub-populations in Xuzhou area so as to develop effective tobacco control policies. Methods Through multi-stage randomized cluster sampling,a face-to-face study with standard questionnaire was carried out among residents aged above 15 years,from June to December 2008,with descriptive nature. Results Totally,44 686 people,with 21 524 males and 23 162 females at age 15 and over were investigated,including 34 391 of them from rural areas and 10 295 from the urban population. Data regarding rates on ever smoked and current smoking,regular smoking,heavy smoking,and average numbers of cigarettes smoked per day,types of tobacco products smoked,rates on quitting smoking successfully,relapse,passive smoke exposure etc.,were 22.45%,21.40%,15.49%,9.49%,15.09/d,4.68%,5.91% and 14.12% respectively. There were no significant differences in the rates of overall smoking,current smoking and regular smoking among urban and rural residents (P>0.05). However,rates of successfully quitting smoking and passive smoke exposure in the urban areas were higher than those in the rural areas (P<0.05). Rates regarding relapse,heavy smoking and average numbers of cigarettes smoked per day in the rural areas were higher than those in the urban areas(P<0.05). Rates on smoking,current smoking,regular smoking,heavy smoking,relapse and average numbers of cigarettes smoked per day were higher in males than those in females (P<0.05). Rates of quitting smoking successfully and passive smoke exposure were higher in females than those in males (P< 0.05). The major types of tobacco products smoked by people aged 15 years old and above were cigarettes (85.17%),and Chinese pipes (3.24%). Conclusion Smoking was quite common in people from Xuzhou. Our data suggested that health education should be strengthened and sustainable intervention measures be developed and implemented to control the tobacco use in the area.

OBJECTIVETo evaluate the prevalence of c-kit and JAK2 gene mutations in protein tyrosine kinase (PTK) family in adult t(8;21) acute myeloid leukemia (AML) and their implications.

METHODSGenomic DNAs from 78 t(8;21) AML patients were screened for mutated c-kit (mutKIT) in exon 8 and 17 by PCR and sequencing. JAK2 V617F mutation screening was processed by allele-specific PCR.

RESULTS(1) Among 78 t(8;21) AML patients, 27 (34.6%) had muc-kit/JAK2 and 6 (7.7%) had JAK2 V617F, and none had both. (2) Peripheral WBC count was higher in mutKIT/JAK2 V617F patients than in wide-type c-kit/JAK2 (wtKIT/JAK2) patients \[(36.2 +/- 37.7) x 10(9)/L vs (21.7 +/- 21.1) x 10(9)/L\] (P < 0.05). There was no significant difference in hemoglobin level, platelet counts, percentage of blast cell in bone marrow, CD117 expression level, the age of onset and gender between the two groups. Peripheral WBC count was higher in mutKIT patients \[(38.8 +/- 40.7) x 10(9)/L\] than in wtKIT patients \[(22.0 +/- 20.4) x 10(9)/L\] (P < 0.05). (3) Complete remission (CR) rates between patients with mutkit/JAK2 V617F and with wtKIT/JAK2 were similar (69.6% vs 80.0%, P > 0.05), but the 2 year continuous CR (CCR) rate was lower in patients with mutKIT/JAK2 V617F (26.7% vs 56.1%, P < 0.05). However, there was no significant difference in OS between mutKIT/JAK2 V617F and wtKIT/JAK2 patients (34.8% vs 58.6%, P > 0.05).

CONCLUSIONSOccurrence of c-kit and JAK2 gene mutations especially c-kit mutation is common in t(8;21) AML patients, and is associated with higher WBC. These mutations confer higher relapse risk and predict poor prognosis.

Adolescent ; Adult ; Aged ; Chromosomes, Human, Pair 22 ; genetics ; Chromosomes, Human, Pair 8 ; genetics ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Janus Kinase 2 ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Mutation ; Prognosis ; Proto-Oncogene Proteins c-kit ; genetics ; Translocation, Genetic ; Young Adult

BACKGROUNDPeripheral T-cell lymphoma (PTCL) is generally characterized by poor prognosis after conventional chemotherapy. The place for high-dose chemotherapy and autologous stem cell transplantation (ASCT) in these patients is still not clear. In this study, we presented the outcomes of PTCL patients followed these treatments in our centre.

METHODSWe retrospectively analyzed the outcomes of 39 patients with PTCL received the two treatments between 1999 and 2010.

RESULTSThe 3-year overall survival (OS) of 61.9% and 3-year progression free survival (PFS) of 35.7% were observed in the 39 patient. Twenty-one patients received Hyper-CVAD chemotherapy with 3-year OS of 46.2% and 3-year PFS of 27.9%. Eighteen patients received ASCT with 3-year OS of 70.3% and 3-year PFS of 44.2%. Further analysis revealed that patients with elevated lactate dehydrogenase, at least 2 international prognostic index (IPI) points, and extranodal involvement had a poorer outcome compared with the control group.

CONCLUSIONThese findings might suggest that Hyper-CVAD chemotherapy and ASCT could offer a durable survival benefit for patients with aggressive PTCL.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Dexamethasone ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Lymphoma, T-Cell, Peripheral ; drug therapy ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Vincristine ; therapeutic use ; Young Adult

OBJECTIVETo screen and obtain the validate immunogenic membrane antigens in pancreatic cancer.

METHODSPancreatic cancer cell line SW1990 membrane protein was extracted and separated by two-dimensional gel electrophoresis (2-DE). One of the three parallel 2-DE gels underwent Coomassie blue staining while the other two underwent immunoblot. Serum IgG was purified from clinically collected sera of 66 pancreatic cancer patients and 24 chronic pancreatitis patients and used as the primary antibody of the immunoblot. Positive dots of immunoblot were identified by MALDI-TOF mass spectrometry and PMF matching. The candidate membrane antigens were further validated respectively in cell lines and tissues by RT-PCR, real-time PCR, Western blot, and their different expression level of gene and protein between pancreatic cancer cell line and normal pancreatic tissue were compared studied.

RESULTSThe immunoblot of SW1990 membrane protein with serum IgG from cancer patients showed nine positive dots which were not the same as those from immunoblot with serum IgG from chronic pancreatitis patients. One talent dot was identified with MALDI and PMF as VDAC2. RT-PCR and real-time PCR showed that the gene of VDAC2 was expressed in the pancreatic cancer cell line. Western blot showed that the expression of protein level of VDAC2 in the pancreatic cancer cell line was obviously higher than in normal pancreatic tissue.

CONCLUSIONSVDAC2 might be the candidate immunogenic membrane antigens of pancreatic cancer, and its gene is all expressed in the pancreatic cancer cell line SW1990, AsPc and P3. The protein level of VDAC2 is significantly overexpressed in pancreatic cancer cell line than in normal pancreatic tissue.

Adult ; Aged ; Antigens, Neoplasm ; isolation & purification ; Cell Line, Tumor ; Female ; Humans ; Male ; Membrane Proteins ; immunology ; isolation & purification ; Middle Aged ; Pancreatic Neoplasms ; immunology ; Proteomics ; Voltage-Dependent Anion Channel 2 ; metabolism ; Young Adult

OBJECTIVETo detect the expression profile of NK ligands in acute leukemia cell lines and investigate the differential expression pattern between acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).

METHODSUsing quantitative real-time PCR, 23 NK ligands (MICA, MICB, ULBP-1, ULBP-2, ULBP-3, ULBP-4, HLA-E, HLA-G, CD48, NBTA, HLA-F, LLT-1, PVR, Nectin2, CD72, CD80, ICAM-1, LFA-3, CRACC, Fas, DR4, DR5, TNFR1) were detected in 6 acute leukemia cell lines, including 3 ALL cell lines (CEM, Jurkat T, Reh) and 3 AML cell lines (HL-60, KG-1a, NB4), respectively. Independent-samples t test analysis was performed to determine statistical significance.

RESULTSUsing β-actin as reference gene, the relative expression results showed that the expression of 4 NK ligands between ALL and AML is significantly different. Specifically, the level of ULBP-2 is higher in ALL (CEM: 1, Jurkat T: 0.617, Reh: 0.246) than that in AML (HL-60: 0.000, KG-1a: 0.003, NB4: 0.000)(P = 0.047). However, the expressions of CD48, PVR(PVR-1, PVR-2) and DR4 is higher in AML (HL-60: 13.987, 4.403, 10.334, 8.711; KG-1a: 5.387, 2.900, 7.315, 4.512; NB4: 7.763, 3.248, 7.049, 6.127) than that in ALL (CEM: 1, 1, 1, 1; Jurkat T: 2.035, 1.553, 3.888, 0.449; Reh: 1.559, 0.000, 0.000, 1.304) (P = 0.044, 0.014, 0.014, 0.011). And there're no significant differences between the rest 19 NK ligands.

CONCLUSIONSULBP-2, CD48, PVR and DR4 might play an important role in the distinct mechanisms in leukemogenesis between ALL and AML and could be potential targets for diagnosis and treatment.

Acute Disease ; Antigens, CD ; genetics ; metabolism ; CD48 Antigen ; Cell Line, Tumor ; GPI-Linked Proteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Ligands ; Membrane Proteins ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Receptors, Virus ; genetics ; metabolism

This study was purposed to investigate the therapeutic effects of early transfusion of immunized donor lymphocytes after haploidentical transplantation by means of mouse model of nonmyeloablative haploidentical bone marrow transplantation. CB6F1 female mouse was served as recipient and C57BL/6 male mouse was served as donor. Each CB6F1 female mouse was subjected to intravenous transfusion with 1×10(6) erythroleukemia (EL9611) cells at day 4 before transplantation, followed with intraperitoneal injection of Ara-C (0.015 g) respectively at day 2 and day 1, then conditioned for BMT with TBI (450 cGy) at day 1 before transplantation. After conditioning (day 0), each of recipients was transplanted with 6×10(7) mixture of bone marrow and spleen cells from the C57BL/6 mice, and was infused with 6 × 10(7) immunized donor lymphocytes at day 15 after transplantation. All treated animals were evaluated for survival, development of leukemia and aGVHD. The donor CD3(+) cell chimerism and sex determining region Y gene (SRY)in recipients were monitored periodically after transplantation. The results showed tht all mice with only inoculation of 10(6) EL9611 cells survived for 15 ± 1 days (n = 4); all mice of other groups obtained the varying degrees of implantation. SRY could be detected at day 30 and 60 after transplantation. The chimerism of donor CD3(+) cells in mixed bone marrow transplantation (MT) group at day 14, 30 and 60 respectively reached 17.95% ± 12.03%, 37.34% ± 2.78% and 47.06% ± 6.1%. In donor lymphocyte infusion (DLI) group it reached 69.78% ± 12.62%, 75% ± 15.97%, 83.41% ± 16.07% at day 30, 45 and 60 after transplantation. The mice of MT and DLI group survived for 66.66 ± 1.47 days and 78.2 ± 7.82 days. It is concluded that the high tumor burden before transplantation can affect donor cell engraftment and prognosis.Early post-transplanted infusion of immunized lymphocytes from donor can help to improve the therapeutic efficacy and survival.
Animals ; Bone Marrow Transplantation ; methods ; Female ; Haplotypes ; Leukemia, Erythroblastic, Acute ; therapy ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors ; Transplantation Conditioning ; methods ; Transplantation, Homologous

PEGlated interferons have many advantages compared with native interferons including longer half-life,fewer intervals,higher stability of blood concentration and so on,and more and more related products are coming into the market.The difference with native interferons in structure gives PEGlated interferons unique physicochemical and biological characteristics and production processes,as well as its specificity in quality control.Here we focus on discussing the main difficulties in quality control of PEGlated interferons,including modification degree,modified site,molecular weight,and so on,providing reference for the research and development,manufacture,and supervision of related products.

Adult ; Antimetabolites, Antineoplastic ; administration & dosage ; therapeutic use ; Azacitidine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Female ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; Male ; Middle Aged ; Myelodysplastic Syndromes ; drug therapy ; genetics ; Treatment Outcome