In comparison with the American Society of Hematology (ASH) annual meeting last year,more studies of hematologic malignancies using next generation sequencing (NGS) are published in the 56th ASH meeting this year.NGS was mainly used in the diagnosis and classification,risk stratification,MRD detection,clonal evolution,prognosis and therapy guidance.This short report will address the application of NGS in the study of hematologic malignancies by summarizing several representative publications from the 56th ASH meeting.

At the 58th American Society of Hematology Annual Meeting,there were many reports on clonal evolution related with blood disease.Gene mutation and clonal evolution after treatment of azacitidine (AZA) in myelodysplastic syndromes (MDS) were summarized to explore the relationship between clonal evolution and treatment response and clinical process in MDS,and to provide reference for clinical treatment decision.

OBJECTIVETo investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.

METHODSAffymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.

RESULTSCompared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.

CONCLUSIONSHepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.

Carcinoma, Transitional Cell ; genetics ; pathology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Humans ; Nuclear Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proteins ; genetics ; physiology ; Urinary Bladder Neoplasms ; genetics ; pathology

The paper is aimed to establish a methods for identication of pearl powder and conch powder from different origins. Hermetic aluminum pan was used to encapsulate samples. The optimal testing conditions were: heating rate 10 degrees C x min(-1), sample weight 3 mg and nitrogen gas flow rate 40 mL x min(-1). The enthalpy values of pearl powder and conch powder was obvious different. Identication of pearl powder and conch powder by DSC is a practical method for its accuracy, convenience and practificality.
Animal Shells ; chemistry ; Animals ; Calorimetry, Differential Scanning ; methods ; China ; Discriminant Analysis ; Pinctada ; chemistry ; classification ; Powders ; chemistry

OBJECTIVETo study the differences of theantimicrobial-resistant profiles between the isolates of Staphylococci aureu from children and from adults.

METHODSStaphylococci was identified by the plasma coagulase test, Staphylococci monoclonal antibody and VITEK-32 fully automated microbiology analyzer.Antimicrobial susceptibility testing was done by the K-B disk diffusion for 84 Staphylococci isolates from children and 74 Staphylococci isolates from adults. Cefoxitin was used for detecting methillicin-resistant Staphylococcus aureus (MRSA) by the disk diffusion test.

RESULTSSeven (8%) MRSA isolates were found in Staphylococci isolates from children compared with 35 MRSA isolates (47%) in those from adults (p<0.01). All strains were susceptible to vancomycin. All strains from children were susceptible to fusidic acid. The resistant rates of the isolates from children to cefazolin, cefuroxime, gentamicin, cefoxitin, and levofloxacin were significantly lower than those from adults (p<0.01).

CONCLUSIONSThe antimicrobial resistance of the Staphylococcus aureus isolates from adults is more prevalent than that in the isolates from children.

Adult ; Child ; Drug Resistance, Bacterial ; Humans ; Methicillin-Resistant Staphylococcus aureus ; isolation & purification ; Microbial Sensitivity Tests ; Staphylococcus aureus ; drug effects

Objective: To investigate the feasibility of multiplex real-time RT-PCR with fluorescent probes in early screening of Ph-like acute lymphoblastic leukemia (ALL) and analyze the clinical feature and prognos. Method: A total of 118 adult B-ALL patients diagnosed between October 2010 and March 2016 were enrolled in this study. Multiplex RT-PCR was used to detect the Ph-like ALL related fusion gene and CRLF2 expression in 58 BCR-ABL and MLL rearrangement negative patients. The clinical features, treatment response and prognosis were analyzed in Ph-like fusion gene positive and/or CRLF2 over-expression patients. Result: Among 58 patients, 9 patients (9/58, 15.5%) showed Ph-like ALL related fusion genes positive and 10 patients (10/58, 17.2%) showed CRLF2 over-expression. There were statistical differences in age, WBC count, immunophenotypes, cytogenetics and risk stratification among Ph-like fusion gene positive or CRLF2 over-expression patients, Ph⁺ patients, MLL⁺ patients and B-other patients. The 2-year overall survival rates were 65%, 47%, 64% and 74% respectively among these four groups (P=0.043) . The 2-year relapse free survival rates were 51%, 39%, 62% and 70% respectively among these four groups (P=0.010) . Conclusion Routine screening of Ph-like ALL by multiplex RTPCR is feasible.

Fatty acids(FAs),which were initially recognized as energy sources and essential building blocks of biomembranes,serve as the precursors of important signaling molecules.Tracing FA metabolism is essential to understanding the biochemical activity and role of FAs in physiological and pathological events.Inspired by the advances in click chemistry for protein enrichment,we herein established a click chemistry-based enrichment(CCBE)strategy for tracing the cellular metabolism of eicosapentaenoic acid(EPA,20:5 n-3)in neural cells.Terminal alkyne-labeled EPA(EPAA)used as a surrogate was incubated with N2a,mouse neuroblastoma cells,and alkyne-labeled metabolites(ALMs)were selectively captured by an azide-modified resin via a Cu(I)-catalyzed azide-alkyne cycloaddition reaction for enrichment.After removing unlabeled metabolites,ALMs containing a triazole moiety were cleaved from solid-phase resins and subjected to liquid chromatography mass spectrometry(LC-MS)analysis.The proposed CCBE strategy is highly selective for capturing and enriching alkyne-labeled metabolites from the complicated matrices.In addition,this method can overcome current detection limits by enhancing MS sensitivity of targets,improving the chromatographic separation of sn-position glycerophospholipid regioisomers,facilitating structural characterization of ALMs by a specific MS/MS fragmentation signature,and providing versatile fluorescence detection of ALMs for cellular distribution.This CCBE strategy might be expanded to trace the metabolism of other FAs,small molecules,or drugs.

Objective: To explore the prognostic value of the international prognostic index (IPI), the national comprehensive cancer network IPI(NCCN-IPI)and the age-adjusted IPI (aa-IPI) in diffuse large B cell lymphoma. Methods: A total of 311 patients with de novo diffuse large B-cell lymphoma (DLBCL) diagnosed from 2003 to 2012 in Nanfang hospital were included. All patients were divided into CHOP (cyclophosphamide, vincristine, doxorubicin, and prednisone) and R-CHOP (rituximab, CHOP) groups. Survival analysis was compared among IPI, NCCN-IPI and aa-IPI models. Discrimination of three different prognostic models was assessed using the Harrell's C statistic. Results: A total of 311 patients were analyzed. Among them, 128 patients were treated with CHOP regimen and other 183 patients were treated with R-CHOP regimen. In CHOP groups, both NCCN-IPI (5-year OS: 59.7% vs 26.8%, P<0.001) and aa-IPI (5-year OS: 71.0% vs 25.0%, P<0.001) showed better risk stratification for low-intermediate and high-intermediate group than the IPI (5-year OS: 47.6% vs 36.6%, P=0.003). However, in the patients treated with R-CHOP, NCCN-IPI showed better risk stratification in low, low-intermediate, high-intermediate groups (5-year OS: 96.0% vs 83.0% vs 66.5%, P=0.009). According to the Harrell's C statistic, C-index of IPI, NCCN-IPI and aa-IPI for overall survival (OS) were 0.546, 0.667, 0.698 in CHOP group and 0.611,0.654, 0.695 in R-CHOP group respectively. In patients younger than 60 years old, C-index of IPI, NCCN-IPI and aa-IPI for OS were 0.534, 0.675, 0.698 in CHOP group and 0.584, 0.648, 0.695 in R-CHOP respectively. Conclusion: The NCCN-IPI is more powerful than IPI and aa-IPI in DLBCL patients receiving R-CHOP. aa-IPI is a preferable model in predicting prognosis than IPI and NCCN-IPI in anthracycline-based chemotherapy without rituximab.
Antibodies, Monoclonal, Murine-Derived ; Antineoplastic Combined Chemotherapy Protocols ; Cyclophosphamide ; Doxorubicin ; Humans ; Lymphoma, Large B-Cell, Diffuse ; Middle Aged ; Prednisone ; Prognosis ; Retrospective Studies ; Vincristine

OBJECTIVE To explore the effect and mechanism of Bushen Zhuyun formula(BSZYF)on gonadotropins and gonadotropin-releasing hormone receptor(GnRHR)signaling transduction in mifepristone-induced LPD rats,and to find the scientific evidence of"heart,kidney and uterus axis"theory of TCM.METHODS SD female rats were administered mifepris-tone suspension via oral gavage,followed by pharmaceutical intervention.Then,the serum follicle-stimulating hormone (FSH)and luteinizing hormone(LH)levels were detected by ELISA assay,the FSHβand LHβexpression levels in the pitui-tary were determined by qPCR,and the GnRHR expression levels were assessed by qPCR and Western blot analysis.Simulta-neously,transcription factors,such as c-Jun,Elk-1,Egr-1,Nur77 and CREB,and transduction molecules in PKC-MAPK, Ca2+-CAM,and cAMP-PKA signaling pathways,such as PKC,p38MAPK,ERK1/2,CAM,and PKA were assessed by qPCR and Western blot analysis.RESULTS After pharmaceutical intervention,BSZYF increased the serum FSH levels sup-pressed by mifepristone(P<0.05).Moreover,the expression of c-Jun,Elk-1,Egr-1,CREB,PKA,p38MAPK,JNK,ERK and PKA were was significantly upregulated in the BSZYF group compared with that in the mifepristone group(P<0.05). However,there was no statistical difference in Nur77 and CAM expression among the blank,mifepristone and BSZYF groups (P>0.05).CONCLUSION BSZYF can regulate abnormal pituitary gonadotropin secretion and GnRHR signaling transduc-tion systems induced by mifepristone,which confirms that the prescription can regulate the whole function of the reproductive axis,and explains the consistency of"Heart,kidney and uterus axis"theory in TCM and the concept of the hypothalamus-pitu-itary-ovary axis in modern medicine

Objective: To investigate the spectrum of gene mutations in adult patients with B-acute lymphoblastic leukemia (B-ALL), and to analyze the influences of different gene mutations on prognosis. Methods: DNA samples from 113 adult B-ALL patients who administered from June 2009 to September 2015 were collected. Target-specific next generation sequencing (NGS) approach was used to analyze the mutations of 112 genes (focused on the specific mutational hotspots) and all putative mutations were compared against multiple databases to calculate the frequency spectrum. The impact of gene mutation on the patients’ overall survival (OS) and recurrence free survival (RFS) was analyzed by the putative mutations through Kaplan-Meier, and Cox regression methods. Results: Of the 113 patients, 103 (92.0%) harbored at least one mutation and 29 (25.6%) harbored more than 3 genes mutation. The five most frequently mutated genes in B-ALL are SF1, FAT1, MPL, PTPN11 and NRAS. Gene mutations are different between Ph⁺ B-ALL and Ph^- B-ALL patients. Ph^- B-ALL patients with JAK-STAT signal pathway related gene mutation, such as JAK1/JAK2 mutation showed a poor prognosis compared to the patients without mutation (OS: P=0.011, 0.001; RFS: P=0.014,<0.001). Patients with PTPN11 mutation showed better survival than those without mutation, but the difference was not statistically significant (P value > 0.05). Besides, in Ph⁺ B-ALL patients whose epigenetic modifications related signaling pathway genes were affected, they had a worse prognosis (OS: P=0.038; RFS: P=0.047). Conclusion Gene mutations are common in adult ALL patients, a variety of signaling pathways are involved. The frequency and spectrum are varied in different types of B-ALL. JAK family gene mutation usually indicates poor prognosis. The co-occurrence of somatic mutations in adult B-ALL patients indicate the genetic complex and instability of adult B-ALL patients.