Adolescent ; Bone Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Follow-Up Studies ; Glomus Tumor ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Male ; Melanoma ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tibia ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Objective To study the mechanism of tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)-induced apoptosis of glioma U87 cells.Methods Human glioma U87 cells were treated with human recombinant soluble TRAIL(rsTRAIL),and the cell apoptosis was detecmd with flow cytometry with AnnexinV-FITC/PI double staining.Flow cytometry with DiOC6 staining was used to assess the changes in mitochondrial transmembrane potential(△ψm).The relative activity of caspase-3,-8 and-9 Was measmed by colorimetric assay,and the concentration of cytoplasmic cytochrome C(cyt C) determined using enzyme-linked immunosorbem assay.The effects ofcaspase-8 inhibitor(Z-IETD-fmk)on rsTRAIL-induced apoptosis,△ψm,caspase-3,-8 and-9 activities and cyt C concentration were observed. Results RsTRAIL tinle-dependently induced apoptosis and progressive collapse of △ψm in glioma U87 cells,resulting also in caspase-3,-8 and-9activation and elevated cytC concentration.Caspase-8 inhibitor partially antagonized these biological effects induced by rsTRAIL in U87 cells.Conclusion TRAIL initiates a cascade of mitochondrial events by activating caspase-8 and induces apoptosis of glioma U87 cells.

To study the gene polymorphism of HLA-A, B, DRB1 alleles in patients with chronic myelogenous leukemia and to explore the correlation of HLA with chronic myelogenous leukemia, the polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-RSSO) was used to analyze the polymorphism of HLA-A, B, DRB1 alleles of 293 CML Patients and 406 randomized and synchronous blood donors (healthy and unrelated with patients) from Guangdong Han population. The results indicate that the gene frequency of HLA-A*24 in CML group was 15.53% lower than that of control group (22.09%, RR = 0.63, p = 0.005); the gene frequency of HLA-B*13 in CML group was 10.41% higher than that of control group (6.74%, RR = 1.68, p = 0.016). The gene frequency of HLA- DRB1*14 in CML group was 7.51% lower than that of control group (11.89%, RR = 0.58, p = 0.008). The differences were all statistically significant. It is concluded that the gene frequency of HLA-A*24, HLA- DRB1*14 in CML patients is significantly lower than normal people in Guangdong. The gene frequency of HLA-B*13 in CML patients is significantly higher than normal people in Guangdong. Further study is needed to make sure whether HLA-A*24 and HLA- DRB1*14 are protective gene markers for CML acquisition on Guangdong Chinese Han population and whether HLA-B*13 is a gene marker for CML susceptibility on this population.
Adolescent ; Adult ; Aged ; Blood Donors ; Child ; Child, Preschool ; China ; Female ; HLA-A Antigens ; genetics ; metabolism ; HLA-A24 Antigen ; HLA-B Antigens ; genetics ; metabolism ; HLA-B13 Antigen ; HLA-DR Antigens ; genetics ; metabolism ; HLA-DRB1 Chains ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; immunology ; Male ; Middle Aged ; Young Adult

OBJECTIVE: To analyze the related factors affecting the long-term prognosis of acute myeloid leukemia (AML) children with positive RUNX1-RUNX1T1. METHODS: The clinical data of 63 chlidren with positive RUNX1-RUNX1T1 AML treated by BCH-AML 05 regimen in our hospital from January 2010 to December 2015 were collected and analyzed retrospectively. The level of RUNX1-RUNX1T1 was detected at the time of initial diagnosis (T), after the first induction treatment (T), after the second induction treatment (T), after the first consolidation treatment (T), after the second consolidation treatment (T) and after the third consolidation treatment (T). According to the fusion transcript levels of RUNX1-RUNX1T1 the AML children were divided into low-expression group and high-expression group; the threshold values for grouping were 10 copies/10 β-glucuronidase (GUS), 10 copies/10 GUS, 10 copies/10 GUS, 10 copies/10 GUS, 1 copies/10 GUS and 0 copies respectively. The gained data were enrolled in the statistical analysis. RESULTS: 23 cases of 63 children died during the follow-up period, and the median follow-up time of the remaining 40 children were 30.04 (11-60) months. There were statistically significant differences in CD15 positive rate between low-expression group and high-expression group (P<0.05), however, there were no statistically significant differences in sex, age, FAB typing, platelet (Plt) count, hemoglobin (Hb) and white blood cell (WBC) count and the ratio of bone marrow immature cells at T2 between the two groups (P>0.05). Univariate analysis showed that sex, Plt counts at T and fusion transcript levels at T, T and T correlated with the 5-year overall survival rate (P<0.05). Multivariate analysis showed that fusion transcript level >10 copies/10 GUS at T was an independent risk factor for 5-year overall survival rate (HR=2.13, 95%CI: 1.04-7.78)(P<0.05). CONCLUSION The fusion transcript level after the first induction therapy in RUNX1-RUNX1T1-positive AML children is an independent factor influencing the long-term prognosis.
Child ; Core Binding Factor Alpha 2 Subunit ; Humans ; Leukemia, Myeloid, Acute ; Oncogene Proteins, Fusion ; Prognosis ; RUNX1 Translocation Partner 1 Protein ; Retrospective Studies

Objective To analyze the results of detection on influenza A (H1N1) 2009 virus in Beijing from May 2009 to December 2009 and to understand the epidemiologic characteristics during the pandemic period. Methods The study was conducted from the May 1 to December 27,2009. A total of 101 852 throat swab samples were detected with the real-time RT-PCR assay by the Beijing Network Laboratory. Data was statistically analyzed. Results 9843 samples showed influenza A (H1N1) 2009 positive, with an overall positive rate as 9.66%. In terms of the positive rates, they were 2.85% from May to June, 3.32% from July to August and 8.35% from September to October. The peak month fell in November (29.67%) and December (24.33%). The positive rates among the following subpopulations were: 8.40% among the suspected cases, 4.75% among close contact cases, 11.46% among the influenza-like illness cases and 7.33% among the cluster cases with fever. Positive cases mainly fell in age groups 5-14 and 15-24. The ratio of male to female was 1.5:1.Conclusion During the pandemic period of influenza A (H1N1) 2009, positive cases gradually increased during May to November but slowly decreasing in December.

With defined culture protocol, human embryonic stem cells (hESCs) are able to generate cardiomyocytes in vitro, therefore providing a great model for human heart development, and holding great potential for cardiac disease therapies. In this study, we successfully generated a highly pure population of human cardiomyocytes (hCMs) (>95% cTnT(+)) from hESC line, which enabled us to identify and characterize an hCM-specific signature, at both the gene expression and DNA methylation levels. Gene functional association network and gene-disease network analyses of these hCM-enriched genes provide new insights into the mechanisms of hCM transcriptional regulation, and stand as an informative and rich resource for investigating cardiac gene functions and disease mechanisms. Moreover, we show that cardiac-structural genes and cardiac-transcription factors have distinct epigenetic mechanisms to regulate their gene expression, providing a better understanding of how the epigenetic machinery coordinates to regulate gene expression in different cell types.
Cell Differentiation ; Cell Line ; DNA Methylation ; Embryonic Stem Cells ; cytology ; metabolism ; Epigenesis, Genetic ; Gene Expression Profiling ; Gene Expression Regulation ; Gene Regulatory Networks ; Humans ; Myocytes, Cardiac ; cytology ; metabolism ; Transcription, Genetic

Drug clinical trial risk assessment system will transfer the fo-cus of quality control from post review to prevention control, which can more effectively ensure the quality of drug clinical trials.It can also pro-tect the subjects’ rights and interests in a great degree.This article will discuss the establishment of drug clinical trial risk assessment system.

Objective To investigate the effects of ATP-binding cassette sub-family G member 2 (ABCG2) and CYP3A4 gene polymorphisms on the pharmacokinetics of bicalutamide and compare the pharmacokinetic properties of different formulations of bicalutamide (capsules and tablets).Methods The subjects were randomly divided into two groups:a single dose of oral test preparation or reference preparation 50 mg,the high performance liquid chromatography-mass spectrometry was used to determine of plasma concentration of bicalutamide,the Phoenix WinNonlin 6.4 was used to calculate pharmacokinetic parameters,and bioequivalence evaluation.Analysis the effects of ABCG2 gene polymorphisms on the pharmacokinetics of bicalutamide by SPSS.Results The main phannaeokinetie parameters of the bicalutamide in the test and reference preparation were as follows:Cmax were (900.00 ± 159.20) and (902.40 ± 146.10) ng· mL-1;tmax were (26.40 ± 9.60) and (35.30 ± 19.80) h;AUC0-t were (1.96 × 105 ±3.28 × 104) and (2.02 × 105 ±4.84 × 104) ng · h · mL-1;AUC0-∞ were(2.03 × 105 ±3.62 × 104)and(2.11 × 105 ±5.63 × 104)ng · h · mL-1.The 90％ confidence interval of Cmax was 88.93％-111.39％;the 90％ confidence interval of AUC0-t was 85.97％-110.76％,the AUC0-∞ 90％ confidence interval was 85.18％-111.51％.There was a significant difference of AUC0-t between CC,AA and CA group (P ＜0.05) and a significant difference of AUC0-∞ between CC and AA group (P ＜ 0.05).There was significant difference of CL between CC and AA group (P ＜ 0.05).Conclusion The pharmacokinetic properties of the two preparations are similar.

It is known that SMAD specific E3 ubiquitin protein ligase 1 (SMURF1) mediates autophagy through its E3 ubiquitin ligase activity, but the ubiquitinated substrates of SMURF1 need to be further explored. In this paper, the interacting proteins of SMURF1 in THP-1 cells were captured and identified by co-immunoprecipitation (Co-IP) combined with mass spectrometry. It was found that SMURF1 could physically bind to 222 proteins in THP-1 cells, and Adenosine deaminase acting on RNA 1 (ADAR1) had a higher peptide binding score. SMURF1 overexpression vectors were constructed and transfected into HEK-293T cells, then Co-IP and Western blotting assays verified the interaction between exogenous SMURF1 and endogenous ADAR1. qRT-PCR and Western blotting assays were carried out after transfecting SMURF1 overexpression vectors in HEK-293T cells, which identified that overexpression of SMURF1 attenuated the protein levels of ADAR1 (P<0. 05). However, there was no significant difference in the mRNA level of ADAR1. HEK-293T cells with normal and overexpressing SMURF1 were treated with cycloheximide (CHX), respectively, and Western blotting assays showed a shortened half-life of ADAR1 after overexpression of SMURF1 (P < 0. 05). Furthermore, overexpression of SMURF1 increased the polyubiquitination level of ADAR1 as detected by Co-IP and Western blot (P<0. 05). After the proteasome inhibitor (MG132) treatment, the Western blotting assay was performed to demonstrate that the negative regulatory effect of SMURF1 on ADAR1 was weakened after the proteasome degradation pathway was attenuated (P<0. 05). This study shows that SMURF1 interacts with ADAR1, catalyzes the polyubiquitination of ADAR1 and mediates its degradation through the proteasome pathway, which provides a theoretical basis for exploring the various biological functions of SMURF1 by affecting the stability of ADAR1.

Objective:To obtain the information of alkaloids in Evodia rutaecarpa by HPLC-Q-TOF-MS/MS. Method:Inter Sustain-C₁₈ column (4.6 mm×250 mm,5 μm) was used with 0.2% formic acid water-acetonitrile as the mobile phase for gradient elution. The column temperature was 25℃,the volume flow rate was 1.0 mL·min^-1,and the sample volume is 5 μL. The detection wavelength was 245 nm,and the chromatographic effluent was detected and analyzed by using both positive and negative ions. Result:According to molecular ion peaks and secondary mass spectrometry characteristic fragment ions,as well as the mass spectrometry information of reference substances and relevant literature reports,more than 40 major peaks were analyzed,and 21 alkaloids were identified from the methanol extract of E. rutaecarpa, including 10 kinds of indole alkaloids,10 kinds of quinolone alkaloids,and 1 kind of ephedrine. Main types of alkaloids in E. rutaecarpa were basically clarified. And the research found that the alkaloids have a good response mainly in the positive mode. Conclusion:Based on HPLC-Q-TOF-MS/MS technology, high-performance liquid chromatography (HPLC) separation,mass spectrometry determination of molecular mass,pyrolysis data,literature analysis and retrieval were performed to quickly,accurately and comprehensively identify alkaloids in E. rutaecarpa, so as to provide a scientific basis for the further extraction and separation of the chemical constituents of E. rutaecarpa.