Fanconi anemia gene-associated germline predisposition in aplastic anemia and hematologic malignancies / 医学前沿

Whether Fanconi anemia (FA) heterozygotes are predisposed to bone marrow failure and hematologic neoplasm is a crucial but unsettled issue in cancer prevention and family consulting. We retrospectively analyzed rare possibly significant variations (PSVs) in the five most obligated FA genes, BRCA2, FANCA, FANCC, FANCD2, and FANCG, in 788 patients with aplastic anemia (AA) and hematologic malignancy. Sixty-eight variants were identified in 66 patients (8.38%). FANCA was the most frequently mutated gene (n = 29), followed by BRCA2 (n = 20). Compared with that of the ExAC East Asian dataset, the overall frequency of rare PSVs was higher in our cohort (P = 0.016). BRCA2 PSVs showed higher frequency in acute lymphocytic leukemia (P = 0.038), and FANCA PSVs were significantly enriched in AA and AML subgroups (P = 0.020; P = 0.008). FA-PSV-positive MDS/AML patients had a higher tumor mutation burden, higher rate of cytogenetic abnormalities, less epigenetic regulation, and fewer spliceosome gene mutations than those of FA-PSV-negative MDS/AML patients (P = 0.024, P = 0.029, P = 0.024, and P = 0.013). The overall PSV enrichment in our cohort suggests that heterozygous mutations of FA genes contribute to hematopoietic failure and leukemogenesis.

Progress of targeted therapy for chronic myeloid leukemia / 白血病·淋巴瘤

The application of tyrosine kinase inhibitors (TKI) has revolutionarily improved the treatment outcome and survival of patients with chronic myeloid leukemia (CML). The new purpose of the current CML treatment is to achieve a sustained deep molecular biological response to reduce the relapse because of drug resistance and even to achieve treatment-free remission maintenance and cure of the disease. Four kinds of TKI have been approved internationally for the first-line treatment of newly diagnosed chronic-phase CML, and there are constantly new drugs and treatment regimens in development and clinical research. This article introduces the research progress of targeted therapy for CML in combination with the related reports at the 62nd American Society of Hematology Annual Meeting.

Progress of gene expression and gene expression patterns in hematological malignancies / 白血病·淋巴瘤

Disorders of gene expression are closely related to the biological characteristics, treatment response, and prognosis of hematological malignancies. The rapidly-developing transcriptome sequencing and single-cell transcriptome sequencing technologies in recent years provide powerful tools for discovering marker genes and studying gene expression patterns related to disease diagnosis and treatment. This article reviews the related research progress in conjunction with reports at the 62nd American Society of Hematology (ASH) Annual Meeting.

Influences of Surfactants on Contact Angle of Pharmaceutical Excipients / 中国药师

Objective:To investigate the effect of surfactants on contact angle of pharmaceutical excipients and further study the effect of surfactants at different concentrations on the wettability of excipients and the effect of of surfactants and excipients at different proportions on the wettability of tablets. Methods:Different kinds of excipients were selected for tabletting,and surfactant solutions at different concentrations were prepared. The contact angle of the solutions on the surface of the tablets was measured by a contact angle tester. Results:The contact angle between the surfactants below the critical micelle concentrations and the tablets was the smallest and the wettability was the best. The change of the contact angle on the tablets with hydrophilic materials was less than that with hydropho-bic ones for most of the surfactants at the same concentration. Conclusion:After the addition of surfactants,the contact angle of excip-ient tablets significantly decreases, however, the contact angle will be stable after the amount of surfactants reaches a certain ratio, which shows significant reference value for tablet formula design.

Interpretation of laboratory test results in early and late onset severe preeclampsia / 中华检验医学杂志

Objective To analyze the difference of laboratory test results between early-onset and late-onset severe preeclampsia and to investigate their clinical application values.Methods Totally 108 blood samples were collected from patients with severe preeclampsia who were diagnosed according to the Diagnostic Standard of Obstetrics and Gynecology(7th Edition) published by People′s Medical Publishing House,in Shandong Provincial Hospital affiliated to Shandong University from March to November 2016,which consisted of 64 early-onset severe preeclampsia before 34 weeks gestation(early onset group) and 44 late-onset severe preeclampsia after 34 weeks gestation(late onset group).In addition,42 women with normal pregnancies as the control group were selected.General clinical data were collected,and the blood sample was analyzed through detecting Hb,PLT,fibrinogen (FIB),D-dimer,AST,ALT,urea,creatinine (Cr),uric acid,CRP,urine protein.The tested results were analyzed and compared.Flow cytometry was used to analyze the proportion of T helper 1 cells(Th1) and T helper 2 cells(Th2),and the ratio of Th1/Th2 was also calculated.All data and F test were performed by use of statistical software SPSS19.0.Results The pre-pregnancy body mass index(29.55±4.49,30.66±5.13,26.62±3.17,F=9.829,P<0.05),diastolic blood pressure[(105.17±14.46)mmHg(1 mmHg=0.133 kPa),(99.80±12.56)mmHg,(74.36±8.42)mmHg,F=82.088,P<0.05],Hb[(123.22±14.38)g/L,(117.03±16.48)g/L,(112.62±11.24)g/L,F=7.133,P<0.05],urea[(6.56±2.36)mmol/L,(4.51±1.35)mmol/L,(3.04±0.87)mmol/L,F=51.733,P<0.05],Cr[(68.47±18.05)μmol/L,(61.37±14.37)μmol/L,(48.54±8.73)μmol/L,F=23.737,P<0.05],CRP[(7.68±8.76)mg/L,(5.88±6.03)mg/L,(3.56±2.41)mg/L,F=4.735,P<0.05],urine protein[(3.66±0.76)g/L,(2.20±1.05)g/L,(0.19±0.40)g/L,F=249.714,P<0.05]had a statistically significant difference among the early-onset,late-onset and control groups.The flow cytometry results demonstrated that the proportion of Th1 in early-onset group(19.83±3.04)was higher than that in both late-onset (14.49±2.79)and control groups(11.78±1.17),on the contrary,the result of Th2 was much lower(early-onset:1.02±0.12,late-onset: 1.11±0.12,control: 1.56±0.11),there was statistical significance among these three groups(Th1: F=135.110,P<0.05;Th2: F=293.687,P<0.05).Conclusions It′s necessary to real-time monitor the laboratory indicators,such as liver and kidney function,especially the immunologic function indicators for evaluating the disease of early-onset and late-onset severe preeclampsia and personal treatment,and for ensuring the health of mother and fetus and improving the prognostic of mother and fetus.

EBV DNA detection in the diagnosis of Epstein-Barr virus infection realated diseases in children / 中华检验医学杂志

Objective To investigate the clinical significance of Epstein-Barr virus EBV DNA in children with Epstein-Barr virus infection realated diseases.Methods A retrospective cohort study was performed.Totally 222 blood samples were collected from children who were diagnosed as EBV infection in Shandong Provincial Hospital from June 2012 to August 2013.Fluorescent quantitative PCR( FQ-PCR) was used to analyze the EBV DNA in peripheral blood lymphocytes.ELISA was used to analyze the four EBV serology antibodies in the serum.Two groups of tested results were compared.Heart, hepatic impairment and renal function were analyzed through detecting AST, ALT, BUN, CREA, CK, CKMB.The results were grouped by EBV DNA copy number, and then non-parametric test together with correlation analysis was performed using SPSS21.0 analytics software.Results The positive rate of EBV-CA IgM and EBV DNA was 51.35%(114/222) and 72.97% (162/222) respectively, χ2 =24.01, P1 ×106 copies/ml,Ⅱ1 ×105 -1 ×106 copies/ml, Ⅲ1 ×104 -1 ×105 copies/ml, Ⅳ5 × 103 -1 ×104 copies/ml, Ⅴ<5 ×103 copies/ml), and ALT(χ2 =10.14,P<0.05), BUN(χ2 =18.17, P<0.05), CK(χ2 =13.09,P<0.05), CKMB(χ2 =17.93,P<0.01) had a statistically significant difference between each group.Well, the log value of EBV DNA copy number had a positive correlation relationship with AST(r=0.357,P=0.001), ALT(r=0.376,P=0.001), BUN(r=0.329,P=0.000), CK(r=0.235,P=0.035).Conclusions Detection of EBV DNA can be used for the early diagnosis and assessment of process of the EBV infection related disease in children.The detection of liver, kidney function and myocardial enzymes can be used for evaluating the severity of EBV infection.

Relationship between ABCA3 gene polymorphisms and susceptibility with neonatal respiratory distress syndrome / 中国小儿急救医学

Objective To investigate ABCA3 gene polymorphism and its relationship with neonatal respiratory distress syndrome (NRDS) in the Guangxi Zhuang Autonomous Region of China by genotyping and haplotype analysis.Methods Using a tagging single nucleotide polymorphism (tSNP) strategy and TaqMan (r) real-time PCR,we genotyped 4 tSNPs (rs4787273,rs 1 50929,rs 11867129,and rs 17135889) and one additional coding SNP(rs13332514) of the ABCA3 gene in preterm infants with NRDS(NRDS group,n =45) and without NRDS (non-NRDS group,n =45) and subsequently predicted the haplotypes.The minor allele frequency and the haplotype 'distribution were compared between the two groups.Results The minor allele A(0.14 vs.0.05,P =0.046) and genotype AG (0.289 vs.0.111,P =0.035) frequency of SNP rs17135889 in NRDS group were significantly higher than those in non-NRDS group.Totally 6 haplotypes occurred at a frequency ≥0.01,among which,the haplotype TGGAG,depended on rs17135889,was significantly higher in NRDS group than that in non-NRDS group (0.061 vs.0.000,P =0.014).Conclusion The results suggested that SNP rs17135889 of ABCA3 gene might be related to NRDS in preterm population of the Guangxi Zhuang Autonomous Region.Allele A contributes to NRDS susceptibility in preterm infants.

Distribution of surfactant protein A2 haplotype and its association with preterm respiratory distress syndrome susceptibility / 中华实用儿科临床杂志

Objective To study the distribution of surfactant protein A2 (SP-A2) haplotype and its association with preterm respiratory distress syndrome (RDS) susceptibility in a local Chinese cohort.Methods Using population base and case-control study cohorts,genotyping for four single nucleotide polymorphism (SNP) was performed,rs1059046,rs17886395,rs1965707,rs1965708,in 80 term infants,50 preterm infants with RDS (RDS group) and 50 preterm infants without RDS(control group) by using TaqMan (R) real-time polymerase chain reaction.Infants in RDS group and control group were matched according to gender and gestational age.Frequency of each haplotype was compared between preterm infants with RDS and without RDS,term infants and preterm infants without RDS.Results Most common haplotypes in term infants were 1A0,1A5,1A1.In each preterm infants groups with RDS and without RDS,1A0,1 A5,1 A1 were also the most common haplotypes.Among these three common haplotypes,frequency of 1A0 was lower in preterm infants,including RDS group and control group,than that in term infants.No significant difference was found between preterm groups with RDS and without RDS (P ＞ 0.05),neither in preterm infants with gestational age ≥32 or ＜ 32 weeks.Haplotype 1A0 frequency(0.542) in term infants was significantly higher than that in preterm infants ＜ 32 weeks without RDS (0.329) (x2 =6.06,P =0.01).Conclusions SP-A2 haplotype distribution in a local Chinese population shows ethnic characteristics to some extent.Only SP-A2 does not contribute to the susceptibility for preterm RDS.

Association between HCV-RNA and chemiluminescence immunoassay for detecting anti-HCV antibody / 国际检验医学杂志

Objective To analyze the correlation of real-time fluorescent quantitation PCR(FQ-PCR)for detecting HCV-RNA loading and the chemiluminescence immunoassay(CLIA)for detecting anti-HCV antibody.Methods 587 samples of anti-HCV an-tibody positive detected by CLIA were furteher detected HCV-RNA by FQ-PCR.Results Among 587 samples of anti-HCV anti-body positive by the CLIA screening,225 samples were HCV-RNA negative and 362 samples were HCV-RNA positive detected by FQ-PCR,and the positive rate was 61 .67%,moreover,which was positively correlated with the S/CO ratio detected by CLIA.Con-clusion The positive rate of HCV-RNA is positively correlated with the S/CO ratio detected by CLIA.The result of HCV-RNA can be predicted according to the S/CO ratio.

Using recipient free edge fingernails for chimerism analysis following allogeneic hematopoietic stem-cell transplants / 中华检验医学杂志

ObjectiveTo examine whether DNA extracted from free edge fingernails specimens from patient after hematopoietic stem cell transplantation (allo-HSCT) could be used for short tandem repeat (STR) genotyping and chimerism analyzing,and to observe the chimerism status in fingernails after allo-HSCT.MethodsPeripheral blood,bone marrow,oral mucosa and free edge fingernail specimens were collected from 25 patients which allo-HSCT were performed in Beijing Dao-pei Hospital during Jul.2009 to Sep.2011 and their donor.Genomic DNA was extracted and 15 STR loci genotyping and chimerism analysis were performed.For the first group which including 12 patients,pairs of fingernail and oral mucosa specimens were collected within one month after allo-HSCT and were comparative analyzed.For the second group which including 13 patients,chimerism status in fingernail samples were analyzed 3 months or longer after allo-HSCT,and 3 patients underwent repeated testing at different times.ResultsFor the first group,4 oral mucosa specimens showed donor chimerism with varying degrees,but no donor chimerism was detected.in all of 12 fingernail specimens.For the second group,6.7％ to 82.6％ donor chimerism was detected in fingernail specimens in 5 out of 13 patients.For the 3 patients underwent repeated testing,donor chimerism was continued negative in one cases,but continued positive in the other 2 cases.ConclusionsFree edge fingernail samples of patients within one month after allo-HSCT can be used for STR typing and chimerism analysis,and it is better than oral mucosa samples.There are cells in allo-HSCT donor graft can differentiate into skin cells,donor derived skin cells chimerism can be formed and persist in some patients.Med,2012,35:23-26)

A clinical and laboratory investigation of myelodysplastic/myeloproliferative neoplasms / 中华检验医学杂志

ObjectiveTo investigate distingwished clinical and experimental characteristics of the four main subtypes in myelodysplastic/myeloproliferative neoplasms (MDS/MPNs).MethodsMDS/MPNs 53 cases from Provincial Hospital Affiliated to Shandong University,including 24 cases CMML,13 cases aCML,12 cases JMML,4 cases MDS/MPN-U,were analyzed regarding to 2001 WHO classification.Morphology (M) of peripheral and bone marrow blood cells were observed under microscope.FCM was used in immunological(Ⅰ) analyse on blasts and myelomonocytes in peripheral blood and bone marrow.G-banding technique was used in cytogenetic (C)examination.PCR was used in molecular genetic (M) mutation detection.Numeric data,such as mean Hb,WBC,PLT et al,among several groups,were compared using Single-factor analysisof variance.Student-Newman-Keulstestwasuseincomparingmeansof two groups.Proportions,such as percentage of clinical features,immunological and cytogenetic abnormal cases among different groups,were compared using Chi-square test or Fisher exact test.Results( 1 ) In the course of MDS/MPNs,there were 46 cases (86.8％ ) had paleness and fatigue 33 cases (62.5％ ) had palpable spleen.JMML had most fever and enlargement of lymph node (75.0％,75.0％ ),statistically distinguished from CMML ( 12.5％,12.5％ ) (x2 =14.89,17.98,P ＜ 0.05 ).(2) The hemoglobin was ( 83.1 ± 24.6 ) g/L.WBC counts were ( 19.8 ± 8.1 ) × 109/L.PLT counts were ( 158.7 ± 108.2) x 109/L.Immature neutrophils and blasts were found in peripheral blood.(3)JMML and CMML had most monocytes absolute counts among the subtypes (4.25 ±0.76) (3.62 ±0.76).(4) Almost 100％ JMML had monocytes abnormalities.(5)For 15 cases were detected immunological characteres by FCM,13 cases showed abnormalities.(6)For 29 cases of MDS/MPNs had been analyzed chromosome karyotypes and 12 out of them (41.4％) were abnormal,Ph chromosomes and those AML-defining translocations were all negative,+ 8 and 7-involved- karyotypes were more frequent.(7)23 cases were detected molecular genetic features,in which were all negative.BCR/ABL1 and JAK2 V617F mutation were all negative in the 13 cases of aCML.JAK2 V617F mutation was positive in 1 case of MDS/MPN-U.ConclusionsMost MDS/MPNs had paleness and fatigue,light to mild anemia,cytosis,monocytes low grade of blast and immature neutrophils in peripheral blood with dysplasia in bone marrow.JMML seems has more severe clinical features and more distinguishing laboratory characters.Immunological abnormalities and abnormalkaryotypes are found frequently in MDS/MPNs with no statistical differences among the four subtypes.There is no specific molecular abnormals in MDS/MPNs.( Chin J Lab Med,2012,35:832-837)

Antiproliferation Effect of Beta-TrCP Ubiquitin Ligase Mediated BCR-ABL Protein Degradation in Leukemia K562 Cells / 中国肿瘤临床

Objective: The BCR-ABL fusion gene induced by reciprocal translocation of t (9; 22) (q34; q11) plays an important role in the pathogenesis of chronic myeloid leukemia (CML). Using recombinant ade-noviruses carrying the N-terminal oligomerizaton domain (OD) of the BCR/ABL and chimeric ubiquitin ligase β-TrCP, this study was to investigate the effect of the targeted degradation of oncoprotein BCR-ABL by Ubiqui- tin-Proteasome System on the proliferation of leukemia call line K562. Methods: The recombinant adenovirus-es carrying wild-type β-TrCP gene (Ad5β-TrCP-OD-HA), mutational β-TrCP gene (Ad5 A F-TrCP-OD-HA)and green fluorescent protein gene (Ad5GFP)were amplified in 293 calls and co-infected into K562 cells respec- tively. The rates of infection were analyzed by flow cytometry (FCM). Recombinant protein and BCR-ABL ex-pression was detected by Western blot. Cell proliferation was determined by cell counting and methylcellu- cose clonal cell culture. Cell cycle was observed through FCM. Untreated K562 cells were used as blank con-trols. Result: The leukemia K562 cell lines with exogenous recombinant β-TrCP-OD-HA and F-TrCP-OD-HA gene were established. The infection rates in the three groups were over 66.4% and recombinant protein sus-tained to be expressed. Ad5β-TrCP-OD-HA down-regulated the expression of BCR-ABL and inhibited prolifer-ation of K562 cells. FCM showed that the percentage of cells at S phase was decreased to 10.88%±2.42%, while that of cells at G_0/G_1 was increased to 85.6%±5.61%, with a significant difference (P<0.05). No changes were found in the cell cycle in groups of Ad5 △ F-TrCP-OD-HA and Ad5GFP. Conclusion: There is sustained ex-pression of recombinant β-TrCP-OD-HA protein in K562 cells infected by recombinant adenovirus.β-TrCP-OD-HA could inhibit the proliferation and clonogenicity of K562 cells through targeted degradation of oncoprotein BCR-ABL and arresting the progression of call cycle.

Mechanism research and effects of clock gene Per2 in K562 leukemia cells on their proliferation,differentiation and apoptosis / 第三军医大学学报

Objective To investigate the effects of circadian clock gene Period2(Per2)on the proliferation,differentiation and apoptosis of K562 cells and its probable molecular mechanism.Methods The Per2 expression plasmid pcDNA3.1-Per2 and empty control plasmid were respectively transfected into K562 cells with cationic liposome,and the resistant cells stably expressing Per2 gene were obtained by G418 selection.Their morphological changes were observed under light microscope following Wright-Giemsa staining.Trypan blue excluding staining and MTT assay were employed to evaluate cell proliferation.Flow cytometry was performed to analyze cell cycle distribution and cell apoptosis,and electron microscopy was used to detect cell apoptosis.Meanwhile,the expressions of proliferation and apoptosis associated proteins,such as P53,Cyclin B1 and C-Myc,were respectively detected by RT-PCR and Western blot analysis at mRNA and protein level.Results The K562/Per2 cell line stably expressing Per2 gene was screened out.As compared with either the empty plasmid transfected group(K562/empty)or the untreated group(K562/untreated),K562/Per2 cells was smaller in volume and showed no obvious cellular differentiation.Circadian clock gene Per2 could significantly inhibit both growth and proliferation of K562 cells.The percentage of K562 cells in G2/M phase increased [K562/Per2 group(36.1?5.5)%,K562/empty group(12.5?2.9)%,untreated group(9.7?2.3)%,P