Objective To investigate the relationship between genetic polymorphism of methylenetetrahydrofolate reductase (MTHFR) and the risk of childhood acute lymphocytic leukemia (ALL).Methods 45 patients with ALL and a cohort of 45 matched healthy children were included,and DNA was extracted from their peripheral blood.PCR-RFLP was used to determine the genotypes of MTHFR C677T and A1298C.The adjusted odds tatio (OR) and 95 ％ confidence interwal (CI) were calculated using unconditional logistic regression model.Results The frequency of MTHFR 677 CC,CT and TT genetypes were 31.1％ (14/45),51.1％ (23/45) and 17.7 ％ (8/45) in controls and 51.1％ (23/45),40.0 ％ (18/45) and 8.9 ％ (4/45)in ALL,respectively (x2 =7.48,P =0.04).The frequency of MTHFR 677 T allele were 69.9 ％ (31/45) in controls and 48.8 ％ (22/45) in ALL.The MTHFR 677 T allele had an decreased risk in ALL compared with CC genetype (OR =0.4,95 ％ CI 0.21-0.83).The frequency of MTHFR 1298 AA,AC and CC genetypes were 57.8 ％,40.0 ％ and 2.2 ％ in controls and 18.8 ％,44.4 ％ and 6.8 ％ in ALL,respectively (x2 =11.23,P=0.23).The frequency of MTHFR 1298 C allele were 51.1％ (23/45) in controls and 42.2 ％ (19/45) in ALL.No significant association between the MTHFR 1298 polymorphism and the risk of ALL (OR =1.3,95 ％ CI 0.21-0.83).Conclusion MTHFR 677 polymorphism could significantly decrease the risk of developing childhood ALL,whereas MTHFR 1298 don' t significantly affect the risk of ALL.

Objective To investigate the value of interphase fluorescence in situ hybridization(FISH)technique and the detection of fusion gene in the diagnosis of acute myeloid leukemia(AML)M2 and M3 Methods FISH was used to detect the AML1/ETO fusion gene and/or PML/RARα fusion gene in incipient cases including 9 AML-M2, 12 AML-M3 and 10 AML undetermined as AML-M2 or AML-M3 primarily diagnosed by routine morphology though bone marrow,cytochemical staining and immunophenotyping,which can help diagnose and guide clinical therapy.Results 4 of 9 AML-M2 cases were AML1/ETO positive.Among 12 AML-M3 cases,10 were PML/RARα positive.1 case was detected AML1/ETO fusion gene.In 10 untonfirmed M3 or M2,3 case8 showed AML1/ETO,5 showed PMIJRARot fusion gene and the rest showed neither of the genes.Conclusion As a new technique of the molecular genetics,FISH is accurate, rapid and efficient.It would be of significance not only at diagnosis of AML,but also for subsequent clinical decision-making.

Objective To investigate the change in distribution and antimicrobial resistance of pathogens from blood culture of children,and provide a basis for treatment of bloodstream infection.Methods Pathogens isolated from blood culture of hospitalized children between January 2009 and December 2013 were divided into group 2009-2011 and 2012-2013.Distribution and antimicrobial susceptibility of pathogens were analyzed.Results From 2009 to 2013,a total of 48 455 blood specimens were taken for culture,2 730 pathogenic bacteria were isolated,positive rate was 5.63%.The positive rate of blood culture decreased year-by-year (χ2 =415.30,P <0.01 ).Of 2 730 iso-lates of pathogenic bacteria,gram-positive bacteria,gram-negative bacteria,and fungi accounted for 80.37% (n =2 194),18.68%(n=510),and 0.95%(n=26)respectively.The difference between two groups of pathogenic bacte-ria was significant(χ2 =180.334,P <0.001).Susceptibility rates of gram-positive cocci to vancomycin,linezolid and teicoplanin were all 100%,resistance rates of coagulase-negative Staphylococcus and Staphylococcus aureus to cip-rofloxacin,compound sulfamethoxazole and tetracycline all decreased.Susceptibility rates of gram-negative bacilli to imipenem,meropenem and amikacin were all≥97.50%,susceptibility rate of Klebsiella pneumoniae to levofloxacin was 100%;Of cephalosporins,Escherichia coli and Klebsiella pneumoniae had high resistance except ceftazidime and cefepime.Conclusion Distribution of pathogens from blood culture of children in 2009-2013 changed signifi-cantly,pathogens have high resistance to commonly used antimicrobial agents,more attention should be paid to the monitor of pathogens from blood culture and pathogenic antimicrobial resistance.

Objective To detect expression of TEL-AML1 fusion genes in pediatric cases with acute lymphoblastic leukemia(ALL) and discuss the role of reverse transcriptase polymerase chain reaction(RT-PCR)and fluorescence in situ hybridization(FISH) in detection of t(12 ;21) and the clinical significance. Methods TEL-AML1 fusion gene was identified in bone marrow munonuclear cells from 31 newly diagnosed childhood ALL patients by NRT-PCR, FISH and conventional cytogenetic analysis (CCA). Results TEL-AML1 fusion gene was found in 7 out of 31 cases, accounting for 22.6 % in pediatric ALL, and 7 out of 31 cases accounting for 25.9 % in B-ALL Seven cases were found with t (12;21) by FISH and NRT-PCR. The incidence of the t(12;21) was 22.6 % in newly diagnosed pediatric ALLs. Conclusion It is concluded that TEL-AML1 rearrangement is a frequent molecular abnormality in childhood ALL. t(12;21) is the most common cytogenetic translocations in Chinese pediatric ALLs, but it is always difficult to identify by routine CCA.Other molecular methods, e.g. NRT-PCR and FISH are powerful in detecting such a critical genetic translocation.

Objective To study the over-expression and clinical implications of the oncogene MDM2 in acute leukemia (AL). Methods The expression of MDM2 gene in 100 patients with newly diagnosed and relapse or refractory AL and 20 healthy as control was measured by relative quantitative reverse transcriptase polymerase chain reaction (RT-PCR),then the results was measured by χ2-test,t-test and one-way ANOVA to compare expession positive rate and intensity of MDM2. Results Among 100 patients,fifty-eight had the high expression of MDM2 gene (58 %). The expression level of MDM2 gene in patients was higher than that of health controls(P ＜0.05). The expression positive rate of MDM2 is higher in poor outcome group (67.9 %,19/28)than that in general outcome group (33.9%,19/56) (P＜0.05). Conclusion Our results suggest that the expression of MDM2 gene plays an important role in the pathogenesis and poor outcome of AL.

Objective To analyse the fusion genes derived from chromosome structural aberrations in acute myeloid leukemia(AML) and the relationship between fusion genes and the MICM classification, clinical diagnosis, chemotherapy and prognosis. Methods The expression of fusion gene in bone marrow samples was detected with multiplex RT-PCR technique and chromosome karyotypes, immunological phenotypes and clinical data were analyzed in 60 acute myeloid leukemia newly diagnosed. Results 37 cases(61.67 %) of 60 patients carried 5 kinds of fusion genes consisting of MLL-AF9, TLS-ERG, CBFβ-MYH1, AML1-ETO and PML-RARα. The activation of oncogene HOX11 was detected in 13 AML cases, three of them with other chromosome aberration simultaneously.23 cases of 31 patients carrying AML1-ETO or PML-RARα, reached complete remission(CR) after chemotherapy and without relapse. Conclusion Gene typing is the most precise classification method that can direct clinical treatment and evaluate prognosis. Multiplex RT-PCR technique, which can quickly screen 29 kinds of fusion gene derived from chromosome structural aberrations at one time, maybe helpful to improve M1CM classification and guide the choice of treatment.

Objective To observe the effect of aspirin on phtelets activation markers in patients with coronary heart disease and set up a diagnostic criteria of aspirin resistance.To preliminarily predict the incidence of aspirin resistance in hospital patients.Methods The subjects were divided into 3 groups:aspirin group(103 cases),control group(24 eases),and healthy control group(23 cases).Using whole blood samples,we detected the ratio of CD62P,PAC-1 expression by flow cytometry(FCM)before and after 7-day treatment and compared the changes of CD62P and PAC-1 expression ratio,then calculated the inhibition ratio of platelets glycoprotein,set up the diagnostic criteria of aspirin resistance with receiver operator characteristic curve(SOC)and calculate the incidenee of aspirin resistance in hospital patients.Results The statisticsl reaults are listed as below:in asptirin group,before treatment CD62P(10.16±6.80)%,PAC-1(14.66±10.56)%,and after treatment CD62P(5.70±4.28)%,PAC-1(8.93±7.08)%,P＜0.01.In control group,before treatment CD62P(9.14±6.52)%,PAC-1(17.67±11.53)%,and after treatment CD62P(7.81±5.72)%,PAG-1(14.97±8.05)%,P＜0.05.According to ROC,the inhibition ratio of CD62P＜21.5% or PAG-1＜17.7% was individually set up asdiagnostic criteria of AR.Our study indicate that the incidence of aspirin resistance in hospital CHD patients is 17.5%.Conclusion There exists platelet activation in CHD patients.CD62P and PAC-1 could be considered as the sensitive index of platelet activation and used in the evaluation of anti-platelet therapy.Aspirin can decrease the expression of CD62P and PAC-1,and inhibit the activation of platelet.According to this study,aspirin resistance really exists in CHD patients.By FCM,the diagnostic criteria of aspirin resistance in CHD is the inhibition ratio of CD62P＜21.5% or PAC-1 ＜17,7% due to aspirin.The incidence of aspirin resistance in hospital CHD patients is 17.5%.

ObjectiveTo investigate the effect of acute normovolemic hemodilution (ANH) on the apoptosis in hippocampal cells induced by global cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy 50-60 day old male SD rats weighing 280-320 g were randomly divided into 3 groups ( n =12 each):group sham operation (group S); group global cerebral I/R (group I/R) and group ANH.Global cerebral I/R was produced by 4-vessel technique described by Pulsinelli et al.in groups I/R and ANH.ANH was carried out at 24 h after cauterization of bilateral vertebral arteries,before occlusion of bilateral carotid arteries.Blood was withdrawn from femoral artery until Hct was reduced to 30％ and equal volume of hydroxyethyl starch 130/0.4 sodium chloride was infused into femoral vein simultaneously.Bilateral carotid arteries were blocked for 5 min at 10 min after ANH.The rats were sacrificed at 24 h of reperfusion and their hippocampi were isolated.Apoptosis was detected by flow cytometry.The expression of Apaf-1 mRNA and caspase-3 mRNA was determined by RT-PCR.Results Global cerebral I/R significantly increased apoptosis index and up-regulated Apaf-1 mRNA and caspase-3 mRNA expression in group I/R as compared with group S.ANH significantly attenuated apoptosis and down-regulated Apaf-1 mRNA and caspase-3 mRNA expression in group ANH compared with group I/R.ConclusionANH can reduce hippocampal cell apoptosis induced by cerebral I/R through down-regulation of Apaf-1 and caspase-3 expression in hippocampus.

Objective To explore the DNA expression of human cytomegalovirus (HCMV) in glioma and the association between HCMV infection and prognosis of glioma patients.Methods Used for this study were 89 specimens of glioma which had been surgically ablated and pathologically confirmed from the patients between January 2007 and December 2016 at Department of Neurosurgery,The First Affiliated Hospital of Zhengzhou University,and Department of Neurosurgery,The First Hospital of China Medical University.Of them,32 belonged to WHO grade Ⅱ,31 to WHO grade Ⅲ and 26 to WHO grade Ⅳ.Ten specimens of normal brain tissue were excised as controls from the contemporary patients receiving resection for essential epilepsy.Nested PCR was used to analyze the DNA expression of HCMV in the glioma tissue and normal brain tissue,and in the peripheral blood from the glioma and control patients.Prognosis of the glioma patients was evaluated using the Kaplan-Meier survival analysis.Results The DNA expression of HCMV was positive in 46 of the 89 specimens of glioma,involving 14 cases of WHO grade Ⅱ,16 ones of WHO grade Ⅲ and 16 ones of WHO grade Ⅳ.The DNA expression of HCMV was negative in all the 10 specimens of normal brain tissue.There was a significant difference in the DNA expression of HCMV between the glioma tissue and normal brain tissue (P=0.002).The HCMV DNA was measured in the peripheral blood from 26 glioma patients,involving 10 cases of WHO grade Ⅱ,8 ones of WHO grade Ⅲ and 8 ones of WHO grade Ⅳ.No HCMV DNA was detected in the peripheral blood from the 10 control patients.There was a significant difference between the brain glioma and control groups in gene expression of HCMV in peripheral blood (P=0.048).There were no significant differences in the survival rate between the patients with positive or negative DNA expression of HCMV in the glioma tissue or in the peripheral blood from the glioma and control patients (x2=1.849,P=0.174;x2=0.082,2=0.774).Conclusion HCMV infection may play an active role in pathogenesis and development of glioma.

OBJECTIVE: To explore the genetic basis for a child with unexplained global developmental delay (GDD), seizure, and facial deformity. METHODS: Whole exome sequencing (WES) was carried out for the patient. Candidate variants were verified by Sanger sequencing of the patient and his parents. RESULTS: WES revealed that the patient has carried a previously unreported de novo heterozygous nonsense c.4906C>T (p.Arg1636Ter) variant of the KMT2A gene, Based on the American College of Medical Genetics and Genomics standards and guidelines, the c.4906C>T variant of KMT2A gene was predicted to be pathogenic (PVS1+ PS2+ PM2+PP3). CONCLUSION The heterozygous nonsense c.4906C>T (p.Arg1636Ter) variant of the KMT2A gene probably underlay the disease in the child. Above finding has enriched the spectrum of pathogenic variants of the KMT2A gene.
Abnormalities, Multiple/genetics* ; Child ; Histone-Lysine N-Methyltransferase/genetics* ; Humans ; Intellectual Disability/genetics* ; Male ; Myeloid-Lymphoid Leukemia Protein/genetics* ; Syndrome