Objective To study telomere dynamics in bone marrow from patients with leukemia and myelodysplastic syndromes (MDS) by flow cytometry and FISH (FLOW FISH) Methods By FLOW FISH , we detected telomere lengths of leukocytes in bone marrow from 43 leukemia patients and 48 healthy donors The mean fluorescence intensity of cells (Q FISH) was detected by flow cytometer Results Compared with the age matched normal range (0 066?0 015), 38 of 43 patients had shortened telomere length (0 047?0 019), and the remaining patients (3 MDS and 2 chromic phase CML) had normal telomere lengths (0 069?0 007 vs 0 060?0 008) Although no significant difference in telomere lengths could be demonstrated in 8 leukemia patients at diagnosis and follow up, a trend toward telomere shortening was observed in CLL patients with 0 052 vs 0 048 in which telomere tended to shorten as the disease went on In contrast, longer telomeres were found in AML patients after induction chemotherapy (0 056) compared with those found in diagnostic specimens (0 043) In the healthy donors, the telomere length turned to be shorter with increase of age ( r =0 67), whereas in leukemia and MDS patients, the telomere length had no correlation with age ( r =0 14) Conclusion The telomere lengths from most patients with leukemia and MDS were shorter than that of healthy donors The telomere dynamics is associated with prognosis and therapy of disease

OBJECTIVE To investigate the bacterial distribution and the extent of drug resistance in lower respiratory tract disease among aged people.METHODS From Jan 2006 to Dec 2006 we collected 304 sputum samples of the old patients who were diagnosed lower respiratory tract infection,bacterial distribution and antibiotic resistance of 178 strains of the pathogens were analyzed retrospectively.RESULTS The most common four kinds of Gram-negative bacilli were Pseudomonas aeruginosa,Klebsiella pneumoniae,Escherichia coli and Acinetobacter baumannii.For above bacteria,the ratio of the resistance to ampicillin,cefazolin and compound sulfamethoxazole was over 40%.The isolating rate of ESBLs-producing E.coli and K.pneumoniae was 29.8%.CONCLUSIONS Monitoring the common bacteria and ESBLs bacteria precisely can provide therapy,guide to early cure and control lower respiratory tract infection of aged patients effectively.

Objective To establish a quality control for Tianmatoufengling Capsule. Methods HPLC method was taken for the determination of Gastrodine in Tianmatoufengling Capsule,A Kromasil C18 column was used,with methanol 0.05% Calcium phosphace(2.5∶97.5) as the mobile phase,and its flow rate was 1.0 mL/min. The detection wavelength was 220 nm. TLC method was used for the identification of Tianmatoufengling Capsule. Results Under the used chromatographic condition,Tianmatoufengling had fine linear responses(r =0.999 9) in the concentration range of 6.016～96.256 ?g/mL,the average recovery was 100.02%,RSD was 2.24%. The method used in TLC-qualitative analysis was simple and sensitive with high specificity,four ingredients identified in Tianmatoufengling Capsule could be detected and all showed specific spots. Conclusion The method employed in this study was convenient and reliable with good repetition. It can be used for quality control in production of Tianmatoufengling Capsule.

Objective To analyze the diagnostic process of a rare case of acute myeloid leukemia with minimal differentiation undergoing a lineage switch to mixed phenotype acute leukemia, NOS-rare types,and to investigate its difference from other acute myeloid leukemia and mixed phenotype acute leukemia. Methods Following tests were performed on the patient with switched mixed phenotype acute leukemia and three control leukemia patients ( including two acute myeloid leukemia with minimal differentiation and one mixed phenotype acute leukemia ). Cell morphology was analyzed by bone marrow smear and related cell chemical staining. Immunophenotyping of bone marrow was performed by flow cytometry ( FCM ). G-banding technique was used for karyotype analysis and RT-PCR was used for fusion gene detection. All the laboratory data of the switched patient were compared to that of three control patients in order to reveal the characteristics of such a rare phenotype switch in acute leukemia. Results Before switching, the morphology of acute myeloid leukemia with minimal differentiation demonstrated 0.82 blasts occurring in bone marrow, distinct nucleoli and absence of Auer rods. Blast cells expressed hematopoieticassociated antigens ( CD38, HLA-DR ), myeloid antigens ( CD13, CD56, CD11b ) and CD7. And these blasts were negative for MPO, CD33, CD15, CD79, CD19, CD22, cytoplasmic CD3, CD4 and CD8. After switching, 0. 42 blasts were found in bone marrow, showed eosinophilia and presence of basophile. Blast cells expressed hematopoietic-associated antigens ( CD38, HLA-DR ), myeloid antigens ( MPO, CD13 ),lymphoid antigens ( CD19, CD79a ,cytoplasmic CD3, and CD7 ). The control group showed typical morphology and immunophenotyping. No abnormal karyotype and fusion gene were detected. Conclusions It is a rare and complicated case that acute myeloid leukemia with minimal differentiation switched to mixed phenotype acute leukemia, NOS-rare types. The laboratory features, especially the change of immunophenotyping play an important role in the diagnosis.

Objective To inhibit the expression level of SALI4 in AML cell line THP-1 and investigate its potential effects on pathogenesis of leukemia. Methods AML cell line THP-1 was transfected with plasmids that expressed small interfering RNA targeting SALL4. The samples were divided into 4 groups:(1) blank group: samples with not any treatments; (2) control group: cells with empty pRS vector alone;(3) test1 group:cells with SALL4-shRNA-pRS-1 plasmid transfection complex; (4) test2 group:cells with SALL4-shRNA-pRS-2 plasmid transfection complex. The expression levels of SALL4 mRNA and protein were measured by real time fluorescence quantitative PCR and WB. C-myc, Cyclin D1 and β-catenin were important components of Wnt/β-catenin signaling pathway and their expression levels in SALL4 knockdown THP-1 cells were detected by real-time fluorescence PCR. Furthermore, THP-1 apoptosis was analyzed by flow cytometry after Annexin V-PI staining. Results Real time fluorescent quantitative PCR illustrated that the expression of SALL4 in testl group, test2 group, control group and blank group were ( 36. 0 ± 4. 3 ) %,(32. 0 ± 2. 4) %, ( 102. 0 ± 6.5 ) % and ( 100. 0 ± 2. 6 ) % respectively. There was statistical significance ( F = 226. 3, P < 0. 05 ). The expression of SALL4 in testl and test2 group respectively were significant lower than that in blank group (t = 19.7,19. 1, P<0. 05). The expression of SALL4 had no significant difference between blank group and control group (t = 1.1, P ＞0. 05). Western blot analysis revealed SALL4 protein in testl and test2 group were significantly decreased compared with those of control and blank group. All above data indicated the high efficiency of RNA interference targeting SALL4. Comparing with the blank group, the relative expression of C-myc, Cyclin D1 and β-catenin mRNA in test1, test2 and control group were(44.0 ±6.2)%,(44.0 ±5.1)% and (107.0±13.6)%;(22.0±4.5)%,(25.0±3.5)% and (48.0 ± 7. 6 ) %; ( 42.0 ± 3.5 ) %, ( 59. 0 ± 3.7 ) % and ( 79. 0 ± 5.6 ) %. The expression of C-myc,β-catenin and Cyclin D1 mRNA in testl and test2 group were significant lower than that in blank group (t = 10. 1,9. 5, 23. 3, 22. 9; 17.4, 12. 4; P < 0. 05). The percentage of apoptotic cells in group of test1,test2,control, blank were (57.2 ±9.1)%, (34.4 ±8.6)%, (14.4 ±3.6)% and (14.8 ±4.8)%respectively. There was statistical significance ( F = 42. 5, P < 0. 05 ). After the inhibition of SALL4, the percentages of apoptotic cell in testl and test2 group were significantly increased( t =9. 7, 4. 5 ;P <0. 05).Conclusion The inhibition of SALL4 in leukemia cell line THP-1 downregulates the expression of cell proliferation related genes such as C-myc, Cyclin D1,β-catenin and promoted apoptosis.

Objective To explore the effects of music therapy and massage on the quality of newborn hearing screening. Methods One hundred and twenty neonates were equally divided according to their registration number: those with even numbers in the experiment group and those with odd number in the control one. Both groups were treated with music therapy. In the experiment group, massage was added by specially trained nurses on the basis of music therapy. The two groups were compared in terms of neonatal hearing test and screening results. Result In the experiment group, the excellence rate of neonatal hearing test and the success rate of screening were significantly higher than those of the control group ( P<0 . 05 ) . Conclusion The music therapy and massage in the neonatal hearing screening can improve the quality of newborn hearing screening.

Objective To study the prevention and corresponding measures for the allergy of contrast agent in scanning, and discuss the targeted prevention mode.MethodsSelection from June 2015 to August 2016, 400 cases was conducted in the people's hospital of ningbo yinzhou MR enhancement scanning patients as the research object, in the process of the study were randomly divided into control group and experimental group, 200 cases in each group.In the control group, patients were given routine procedure, including psychological counseling, diet control and health, education, etc.All patients were treated with the use of gadolinium bate as a contrast agent and intravenous injection.Compare two sets of contrast agent allergy.ResultsTwo groups of patients showed different levels of allergic reactions.The control group had five mild allergies, one moderate allergy, one severe allergy, and seven cases of allergy.There were two cases of mild allergy, 1 case of moderate allergy, no severe allergies, and 3 cases of allergies.Compared with the experimental group, the difference was statistically significant (P<0.05).ConclusionIn MR enhancement scanning to psychological counseling, diet, health education and other measures, to reduce the probability of contrast media in patients with allergic, have clinical significance.

Objective To explore the role of hMSH2, a novel endogenous tumor-associated pro-tein ligand recognized by Vγ9δ2 T cells, in innate anti-cervix cancer immunity .Methods hMSH2 that ex-pressed on the surface of cervical cancer cell line HeLa cells was blocked by specific antibody .Then the differences in their effects on Vγ9δ2 T cells before and after antibody blockage were evaluated by cytotoxicity of Vγ9δ2 T cells and cytokines secretion .The serum levels of hMSH2 in patients with cervical cancer were detected by ELISA .Distribution of hMSH2 in cervical cancer tissues was analyzed by TMA immunohisto-chemistry .Results Decreased cytotoxicity and IFN-γsecretion were observed in Vγ9δ2 T cells against He-La cells blocked with specific anti-hMSH2 antibody .Serum concentration of hMSH 2 in patients with cervical cancer was slightly higher than that of healthy control .Altered distribution pattern of hMSH 2 was observed in cervical cancer tissues .Conclusion hMSH2 is involved in Vγ9δ2 T cells-mediated innate anti-cervical cancer immunity by enhancing their cytotoxicity and IFN-γsecretion.

Objective To investigate the appropriate setting up of normal reference ranges of lymphocyte subsets in some flow cytometry laboratories and to study the effects of different flow cytometers and various reagents by different manufacturers on the analysis of peripheral blood lymphocyte subsets. Methods Three FCM labs (named A, B and C) in Beijing region were selected representing 3 commonly used flow cytometers (Beckman Coulter Epics XL, Beckman Coulter Cytomics FC500, BD FACS Calibur). 50 samples from healthy donors were distributed to 3 labs and tested according to individual lab's standard operating procedure to verify whether the normal reference ranges of peripheral blood lymphocyte subsets established were appropriate. The application of internal quality control was also investigated. Commercial blood quality control reagents were given to the 3 FCM labs and tested within 20 working days paralleled with routine samples. In addition, 20 patients' samples were prepared using 4 different combinations of reagents ( a , b , c and d). The results from combination a, which used the Beckman Coulter reagents and instrument, were compared to the results from combination b, c and d, which used reagents from different manufacturers. Then the prepared samples were tested on Beckman Coulter Epics XL to evaluate the effects of different combinations of reagents on the results of peripheral blood lymphocyte subsets analyzed by the same instrument. Furthermore, 24 patients' samples prepared by same reagents from Beckman Coulter company were tested on both Beckman Coulter Epics XL and BD FACS Calibur respectively to assess the effects of different instruments on peripheral blood lymphocyte subsets. 20 patients' samples prepared by same reagents and instruments were analyzed by Beckman Coulter Epics XL analytic system and BD FACS Calibur analytic system respectively to assess the effects of the two analytic systems on the lymphocyte subsets. Results Over 10% of the results for NK and T4/T8 in lab A as well as T4 in labs B and C fell outside of their normal reference ranges. The probabilities exceeding corresponding normal reference ranges were 16% ( 9/50 ), 24% ( 12/50 ), 22% (11/50) and 12% ( 6/50 ), respectively. The results using internal blood quality control in 3 FCM labs within 20 working days were all within the reference ranges of the quality control provided by the kit. The biases from b and c reagent combinations were substantial compared with that of reagent a combination. Among the biases from b and c reagent combinations, the lowest probability of bias exceeding 10% was T8 of combination b, which had probability of 70% (14/20). The highest probabilities of hias exceeding 10% were T3 and T4 of b and c reagent combinations, which reached 100% (20/20) . Furthermore, the biases of T3, T8 and B of d reagent combination compared with that of reagent a combination were also substantial. The probabilities of bias exceeding 10% were 35% (7/20) ,85% (17/20) and 75% (15/20), respectively. Comparing the results of samples prepared and analyzed by reagents and instruments from different manufacturers to that of samples prepared and analyzed by the same company's reagents and instruments showed that there were great discrepancies in T3, T4 , T8 , B and NK. The probabilities of bias exceeding 10% were 71% ( 17/24), 80% (19/24) ,38% (9/24), 33% (8/24) and 92% (22/24), respectively. The biases of T8, NK and B were substantial when compared the results from Beckman Coulter Epics XL analytic systems and BD FACS Calibur analytic systems. The probabilities of bias exceeding 10% were 55% (11/20 ), 70% ( 14/20 ) and 55% (11/20), respectively. Conclusions FCM labs should set up their own normal reference range for peripheral blood lymphocyte subsets. The normal reference range should be verified periodically. It is important to apply internal blood quality control regularly and accumulate the quality control results. The reagents and instrument for preparing peripheral blood samples should be from the same manufacturers.

Objective To investigate the association of serum angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) with myocardial reperfusion in ST-segment elevation myocardial infarction (STEMI) patients treated with primary percutaneous coronary intervention (PCI). Methods A total of 103 consecutive STEMI patients who received primary PCI were enrolled in this study. The patients were divided into two groups according to ST-segment resolution (STR) at 2 h after PCI:STR ≥ 50%group (n=69) and STR<50%group (n=34). Serum concentrations of Ang-1, Ang-2 and Ang-2 to Ang-1 ratios (Ang-2/Ang-1) before and immediately after PCI, 2 h, 6 h, 24 h after PCI were compared. Predictors of poor STR were identiifed by multivariable logistic regression analysis. Results The patients with STR≥50%had significant higher serum Ang-1 levels (P < 0.05) and lower Ang-2/Ang-1 ratios (P < 0.01) from before PCI to 6 h after PCI than those with STR < 50%;Ang-1 and Ang-2/Ang-1 at 24 h after PCI, and Ang-2 at all time points were not signiifcantly different between the two groups (P>0.05). In multivariable logistic regression analysis, Ang-2/Ang-1 before PCI was independently associated with STR < 50%;Other independent predictors were pain to balloon time, infarct related artery (LAD), and TIMI flow grade<Ⅲafter PCI. Conclusions Higher Ang-2/Ang-1 is an independent predictor of poor myocardial reperfusion in STEMI patients after PCI.