OBJECTIVETo investigate the susceptibility of von Willebrand factor (VWF) type 2A mutant A1500E to proteolysis by metalloprotease ADAMTS13 and to provide the direct supports for the pathogenesis of VWF mutation A1500E responsible for von Willebrand disease (VWD) type 2A.

METHODSRecombinant wild-type VWF (WT-VWF) and A1500E mutant VWF transiently expressed on transfected HeLa cell lines. Expression media were collected and concentrated, then cleaved directly by recombinant ADAMTS13 (rADAMTS13). Compared with WT-VWF, the susceptibility of A1500E mutant VWF to proteolysis by ADAMTS13 was analyzed using SDS-agarose gel VWF multimers analysis.

RESULTSIn vitro the expression of VWF:Ag in the supernatants of WT-VWF and A1500E mutant VWF were 1.10 U/ml and 0.78 U/ml, respectively, while VWF:Ag in cells lysates of A1500E mutant VWF was 90.6% of that of WT-VWF. The SDS-agarose gel VWF multimers analysis showed that there were no differences between WT-VWF and A1500E mutant VWF. The A1500E mutant VWF could be efficiently cleaved by ADAMTS13 under static condition without denaturants such as urea and guanidine HCl. VWF multimeric analysis showed that high and intermediate molecular weight multimers dramatically decreased while low molecular weight multimers obviously increased. Conversely, WT-VWF could not be cleaved by ADAMTS13 under the same condition.

CONCLUSIONThe A1500E mutation resulted in VWF more susceptible to ADAMTS13-dependent proteolysis, which belonged to VWD type 2A group 2 mutation.

ADAM Proteins ; genetics ; metabolism ; ADAMTS13 Protein ; Genotype ; HeLa Cells ; Humans ; Hydrolysis ; Mutation ; Recombinant Proteins ; genetics ; metabolism ; von Willebrand Disease, Type 2 ; genetics ; metabolism ; von Willebrand Factor ; genetics

OBJECTIVETo study the effects of Huangqi decoction (HQD) on phagocytic activity of peritoneal macrophage of mice.

METHODOne hundred Kunming mice, whose weight varied from 18 g to 22 g, were selected and divided into 10 groups randomly in eluding contrast group, groups conducted at different doses of HQD by ig, groups conducted in various ways of taking medicine, and groups conducted with comparative treat combining Huangqi and Dexamethasone. Mice in every group were taken medicine one time daily for 6 days.

RESULTAmong the groups treated at different doses of HQD, phagocytic rate and phagocytic index of mice, which were taken HQD by ig at high, middle, and low doses, were significantly higher (P < 0.01) than that of mice in contrast group, at the same time the effect in group with high dose was the best. Among the groups treated in various ways of taking medicine, phagocytic rate of the ip group was significantly better (P < 0.01) than that of the sc group and that of the ig group respectively, but there was not significant difference (P > 0.05) of phagocytic index among them. Among the groups combining Huangqi and Dexamethasone, Huangqi could antagonize the immunosuppressive effect of Dexamethasone obviously (P < 0.01).

CONCLUSIONHQD at different doses and with various ways of taking medicine could improve phagocytic activity of peritoneal macrophage of mice at different degree, and could antagonize the immunosuppressive effect of Dexamethasone.

Animals ; Astragalus membranaceus ; chemistry ; Dexamethasone ; antagonists & inhibitors ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Macrophages, Peritoneal ; drug effects ; physiology ; Male ; Mice ; Phagocytosis ; drug effects ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation

Objective To investigate the clinical outcomes of patients undergoing microsurgery for cerebellar medulloblastoma. Methods This retrospective analysis of the clinical and follow-up data involves 57 patients who received microsurgery for pathologically confirmed cerebellar medulloblastorna, and the microsurgical techniques for medulloblastoma were discussed. Results Among the 57 patients, 42 had total tumor resection, 13 had subtotal and 2 had partial resection of the tumors. Patency of the midbrain aqueduct was achieved in all the cases after the surgery. Hydrocephalus was found in 43 patients before the operation and only in 16 patients after the operation. Tumor relapse occurred in 19 patients 2 years after the operation, including 8 with implantation metastasis compromising the central nervous system and 1 patient with frontal lobe metastasis who received reoperations. The earliest tumor relapse occurred 20 days after the surgery. The 2- and 5-year postoperative survival rates in these patients were 68.4% and 49.1%, respectively. Conclusion Good therapeutic effects can be achieved with total resection of the tumors and postoperative whole brain and spinal cord radiotherapy in patients with medulloblastomas.

OBJECTIVEThis study investigated the impact of metabolic syndrome on the development of cardio-cerebral vascular (CVD) events in a pre-hypertensive population.

METHODSThe data used in this prospective study was derived from the Kailuan study cohort (n = 101 510). Prehypertension was diagnosed in 29 968 (mean age: 50 ± 9 years and 23 744 males) individuals by the JNC VII criteria and these subjects were further classified into metabolic syndrome positive (MS+, n = 3447) and MS negative (MS-, n = 26 521) groups according to the modified 2004 Chinese Diabetes Society criteria. Subjects were followed up for 38 - 53 (mean 47 ± 5) months and first-ever CVD events were recorded. Baseline anthropometric and laboratory features were obtained by physical examination from June 2006 to October 2007 and the last follow-up day was December 31, 2010. Multivariable Cox proportional hazards regression models were used to analyze the risk factors of first-ever CVD events.

RESULTSThere were 354 CVD events during follow up. The incidences of CVD events (1.80% vs. 1.28%) and cerebral infarction (1.10% vs. 0.57%) were significantly higher in the MS+ group than in the MS- group (all P < 0.05). After adjustment for other established CVD risk factors, the hazards ratio was 1.45 (95%CI: 1.10 - 1.92) for total CVD events and 1.84 (95%CI: 1.27 - 2.67) for cerebral infarction events in MS+ group.

CONCLUSIONSIn this cohort, metabolic syndrome is linked with increased risk for CVD events.

Adult ; Cardiovascular Diseases ; etiology ; Cerebrovascular Disorders ; etiology ; Cohort Studies ; Female ; Humans ; Male ; Metabolic Syndrome ; complications ; Middle Aged ; Prehypertension ; complications ; Prospective Studies ; Risk Factors

OBJECTIVETo evaluate the effectiveness and the practicability of the Acute Lymphoblastic Leukemia Berlin-Frankfurt-Münster 95 (ALL-BFM 95) protocol in treating childhood high-risk acute lymphoblastic leukemia (HR-ALL).

METHODSA retrospective analysis of 47 children with newly diagnosed HR-ALL between July 2003 and August 2013 was performed. These children were treated by the ALL-BFM 95 protocol. Survival was evaluated by Kaplan Meier analysis and Log-Rank test.

RESULTSRelapse-related death occurred in 12 of 47 patients (26%), and 5 of 47 patients (11%) were treatment-related mortality. Five-year probability of event-free-survival (pEFS) was 62%. Children with hematopoietic stem cell transplantation (HSCT) after chemotherapy achieved significantly better pEFS than those with chemotherapy alone (77% vs 52%; P=0.035). The patients who were only poor responders to prednisone had a better outcome (5-year pEFS 80%) than the Days 15 and 33 bone marrow M3 subgroups (5-year pEFS 60% and 0 respectively).

CONCLUSIONSALL-BFM 95 protocol can improve the outcome of children with high-risk ALL. The major cause of death is attributed to relapse. Chemotherapy plus HSCT can produce a better outcome than chemotherapy alone. The Days 15 and 33 bone marrow M3 subgroups have a poor prognosis.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; mortality ; Risk ; Treatment Outcome

OBJECTIVETo investigate the association between the anti-atherosclerotic effects of amlodipine and angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism in elderly essential hypertensive (EH) patients.

METHODSA total of 220 EH patients were treated with amlodipine (2.5 - 10 mg, once daily) for twelve months and complete data were obtained from 208 patients with genotypes of II (n = 90), ID (n = 91) and DD (n = 27). The indices of carotid arterial were compared before and post amlodipine treatment in patients with identical genotype and among different ACE genotypes and each genotype post therapy.

RESULTSThe carotid mean intimal-medial thickness (MIMT) was slightly decreased in EH patients with ID and DD genotypes and significantly decreased in EH patients with II genotype (0.96 +/- 0.12 vs. 0.92 +/- 0.13, P < 0.01) compared to pre-treatment values. The decreased degree of MIMT (DeltaMIMT) in II genotype was significantly higher in II genotype than those in ID or DD genotype (0.05 +/- 0.03 vs. 0.01 +/- 0.02, 0.01 +/- 0.03 respectively, P < 0.01). The post treatment plaque score (PS) in patients with II genotype was significantly reduced (4.85 +/- 2.51 vs. 3.90 +/- 2.36, P < 0.05). Multivariate linear regression analysis showed the baseline SBP, the decreased degree of SBP (DeltaSBP) and the II genotype were the major factors affecting the DeltaMIMT.

CONCLUSIONHypertensive patients carrying II genotype ACE genotype are the best responders for the anti-atherosclerotic effects of amlodipine.

Aged ; Aged, 80 and over ; Amlodipine ; therapeutic use ; Carotid Artery Diseases ; pathology ; prevention & control ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; drug therapy ; genetics ; pathology ; Male ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Genetic ; Treatment Outcome

OBJECTIVETo investigate the chromosomal abnormalities and evaluate the prognostic value of post-remission chemotherapy in children with t (8;21) acute myeloid leukemia (AML).

METHODSThe diagnosis of AML and its subtyping were performed using morphological, immunological, and cytogenetic methodologies in 79 children. Induction therapies included homoharringtonine and cytarabine (HA), daunorubicin and cytarabine (DA), or homoharringtonine and daunorubicin and cytarabine (HAD). Allogeneic stem cell transplantation or 5-6 cycles of intensive chemotherapy was performed after remission therapy.

RESULTSAdditional chromosomal abnormalities, including loss of sex chromosome (n = 40, 50.6%), del (9q) (n = 9, 11.4%), and complex abnormality (n = 7, 8.9%) were identified in 55 patients (69.6%). Three patients had more than 90 chromosomes and duplicate t (8;21) tetraploid karyotype, and their prognoses were poor. The complete remission (CR) rates were 81.7% (49/60) and 94.8% (55/58), respectively, after one and two cycles of induction chemotherapy. The 3-year event-free survival rate (EFS), disease-free survival rate (DFS), and overall survival rate (OS) were (26.2 +/- 6.8)%, (31.3 +/- 6.7)%, and (27.6 +/- 6.6)%, respectively. Twenty-nine patients received 5 or more cycles of chemotherapy after CR and demonstrated an improved 3-year DFS [(51.7 +/- 9.3)%]. The 3-year DFS was not significantly differently in patients with or without additional abnormalities other than sex chromosome (P = 0.36). Post-remission consolidation by high dose cytarabine (HDAC) was significantly superior to standard chemotherapy (66.7% vs. 27.3%, P = 0.03).

CONCLUSIONMost children with t (8;21) AML have additional chromosomal abnormalities, although they do not affect the prognosis and long-term survival. Few patients have more than 90 chromosomes and duplicate t (8;21) tetraploid karyotype, which may result in poor prognosis. Childhood t (8;21) AML usually has high CR rate with relatively good prognosis, and post-remission consolidation by HDAC can improve the survival.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; Chromosomes, Human, Pair 21 ; Chromosomes, Human, Pair 8 ; Female ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; genetics ; therapy ; Male ; Prognosis

To explore the genetic characteristics of viral quasispecies in HIV-1 CRF07_BC infections among intravenous drug users (IDU), the gp120 fragments of HIV-1 env gene were amplified from plasma samples collected from 6 CRF07_BC infected persons using single genome amplification and sequencing (SGA/ SGS) method, and 11 to 28 sequences were obtained from these samples, respectively, A neighbor-joining phylogenetic tree was reconstructed to describe the genetic characteristics of viral quasispecies. The Simplot, segments' phylogenetic trees and diversity plots based on average pairwise distance (APD) were used to identify the recombination events between quasispecies. The SGA sequences derived from single specimen formed a large monophyletic cluster in the neighbor-joining phylogenetic tree and showed the complex topologic structures of viral quasispecies. Of the 6 CRF07_BC infected patients, only one possessed the high genetic homogeneity, whereas the other five individuals showed high heterogeneity, with two to four subclusters inside the monophyletic cluster for each specimen. In addition, the recombinant events were identified among viral quasispecies from 3 cases. The results show SGA technique and phylogenetic analyses are useful tool to investigate the intrahost CRF07_BC gp120 complex quasispecies variation and high genetic diversity.
Adult ; Drug Users ; Female ; HIV Infections ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Substance Abuse, Intravenous ; virology ; Young Adult

OBJECTIVETo characterize the Gag-Specific T lymphocyte responses and identify immunodominant region recognized in Chinese HIV-1 recombinant subtype B/C infectors.

METHODS10 antiretroviral treatment (ART) naive HIV-1 recombinant subtype B/C infectors with infected time in 1 year, 25 ART-naive infectors with infected time > 3 years and 10 HIV-1-seronegative healthy individuals were enrolled. HIV-1-specific T lymphocyte responses were analyzed by an IFN-gamma Elispot assay against 123 overlapping peptides spanning HIV-1 Gag protein in the present study.

RESULTSGag-specific T lymphocyte responses of interferon-gamma secretion were identified in 8(8/10) Chinese HIV-1 recombinant subtype B/ C infectors with infected time in 1 year, the specific T lymphocytes are mainly targeted at five seperated peptides. Responses were identified in 17(68%) infectors with infected time more than 3 years, the specific T lymphocytes are mainly targeted at one peptide in p17 and six in p24. There was obviously positive correlation (P = 0.0318, r = 0.519) between the magnitude of responses and viremia in infectors infected time > 3 years. The magnitude of response in infectors infected in 1 year was significantly higher than group infected time > 3 years (P = 0.021). None of healthy individuals produced positive responses.

CONCLUSIONSHIV-1 recombinant subtype B/C Infectors at different stages of diseases recognize different region of gag.

CD4-Positive T-Lymphocytes ; immunology ; virology ; Gene Products, gag ; genetics ; immunology ; HIV Infections ; genetics ; immunology ; virology ; HIV-1 ; classification ; genetics ; immunology ; Humans ; Immunodominant Epitopes ; Interferon-gamma ; genetics ; immunology ; Recombination, Genetic

OBJECTIVETo investigate the methylation rate of cyclin-dependent kinase inhibitor 2A (CDKN2A) and cyclin-dependent kinase inhibitor 2B (CDKN2B) in the 9P21 region in children with acute myeloid leukemia (AML) and the association of gene methylation with clinical features and outcomes.

METHODSThe clinical data of 58 children who were newly diagnosed with AML between January 2010 and December 2012 were retrospectively analyzed. Thirty-eight healthy children were recruited as the control group. Genomic DNA was extracted from bone marrow or peripheral blood of the 58 patients and 38 healthy children. The methylation status of CDKN2A and CDKN2B was analyzed by methylation-specific multiplex ligation-dependent probe amplification.

RESULTSGene methylation was not found in healthy children. Methylation probes of 44 patients were detected in 58 patients. The methylation of CDKN2A was detected with 136 bp and 237 bp methylation probes. The methylation of CDKN2B was detected with 130 bp, 210 bp, 220 bp, and 417 bp methylation probes. The methylation rate of CDKN2A was 5%, while the methylation rate of CDKN2B was 76%. The methylation detected by some probes was associated with sex, hemoglobin, and platelet count at the first visit.

CONCLUSIONSThe methylation of CDKN2B is a common event in children with AML, while the methylation of CDKN2A is relatively rare.

Adolescent ; Child ; Child, Preschool ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; Female ; Humans ; Infant ; Leukemia, Myeloid, Acute ; genetics ; Male