Objective To explore the relationship between adverse childhood experiences,negative emotions and interpersonal sensitivity.Methods Adverse Childhood Experiences Questionnaire (ACEQ),Depression Anxiety Stress Scale (DASS-21) and the Interpersonal Sensitivity Subscale of Symptom Check-List (SCL-90) were applied to investigate the 492 new enrolled medical students.Results Boys got higher ACEQ scores than girls; Compared with only-child,non-only-child scored higher in depression,interpersonal sensitivity; Compared with students from non-rural areas,those from rural areas got higher scores in depression,anxiety stress,interpersonal sensitivity.There existed significant differences of scores on ACEQ,depression,anxiety,stress,interpersonal sensitivity among children of fathers of different educational level; while differences of scores on depression,anxiety,interpersonal sensitivity among children of mothers of different education level were statistically significant.There were significantly differences of scores on depression,anxiety,stress,interpersonal sensitivity among participants with 0,1,2 and 3 or more ACEQ.The results of correlative analysis provided evidence that adverse childhood experiences,all the factors of negative emotions and interpersonal sensitivity were significantly positively correlative.The SEM suggested that adverse childhood experiences had an indirect effect on interpersonal sensitivity through negative emotions.Conclusions Adverse childhood experiences has a great influence on negative emotions and interpersonal sensitivity.Negative emotions has a completely mediating role between adverse childhood experiences and interpersonal sensitivity.

Objective To prove the clinical effect of portable auto-haemostat machine on body's limbs. Methods Portable auto-haemostat machine was applied to the indicated patients in their clinical aid treatments course, i.e. the rapid hemostasis on limbs. Results The hemostasis was rapid and accurate with hemostatic pressure in the range from 190 to 230 mmHg. The excellent effect guarantees the diagnosis, observation, bandaging and secure transmission of victims until they were sent to the operation room. Conclusion It is reliable to use portable auto-haemostat machine on body's limbs in clinic works.

Objective To analyze the immunophenotyping characteristics of myeloid leukemia in transgenic mouse.Methods According to differential antigen expression profile on various hematopoietic lineages,flow cytometric analysis of bone marrow sample was performed on 5 myeloid leukemia mice and 10 healthy BL6 mice.Cell cycle analysis was further performed to assess cell proliferation.Results Expressions of Mac-1+ Gr-1+ and c-Kit+ in bone marrow cells in transgenic leukemia mice were(72.6±6.5)% and (20.5±4.8)%,and it were significantly higher than those in normal mice[(52.8±4.8)% and(2.1±0.3)%](t=6.66,12.66,P＜0.01).And expressions of B220+,CD3+,CD41+ and Ter119+ in leukemia mice were(2.7±1.1)%,(1.2±0.3)%,(1.2±0.6)% and(2.8±1.1)%,respectively.It were significantly lower than those in normal mice[(20.2±2.1)%.(6.6±1.3)%,(4.7±1.1)% and(10.6±1.2)%](t=-17.63,-8.69,-6.30,-12.28,P＜0.01).The percentages of S phase and G2/M phase in leukemia mice were(25.7±4.2)% and(21.1±4.2)%,respectively.It were significantly increased as compared with normal mice[(11.8±2.1)% and(8.9±1.8)%](t=8.59,7.98,P＜0.01).Conclusions Immunophenotyping of myeloid leukemia in transgenic mouse was characterized by hish expression of myeloid specific marker(Mac-1 and Gr-1)and hematopoietic stem/progenitors cells specific marker(c-Kit),and by low expression of B-lymphoid specific marker(B220),T-lymphoid(CD3),megakaryocyte(CD41)and Erythroid(Ter119).

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.

MicroRNAs (miRNAs) are small noncoding regulatory RNAs ranging in size from 17 to 25 nucleotides which participate many physiological and pathological processes.MicroRNA could also stably exist in peripheral blood,and the detection of circulating microRNA is of great significance for disease diagnosis and prognosis.As so far,there is no unified method and standard for detection of microRNA,which is the major reason for discrepant results between different studies.However,circulating microRNA,as a new disease detection biomarker,has stable properties,convenient detection and high accuracy features,so it till has an important potential value in clinical applications.

Objective To study the clinical significance of a novel marker of platelet clumps count provided by hematology analyzer in differentiating true thrombocytopenia from EDTA-dependent pseudothrombocytopenia (EDTA-PTCP). Methods Samples from 65 cases of thrombocytopenia (including 15 EDTA-PCTP samples and 50 random samples of true thrombocytopenia) and 50 healthy controls were analyzed using hematology analyzers, and samples with low platelet counts were checked by replacing citric acid and using manual microscope observation to identify true thrombocytopenia from EDTA-PTCP. A novel marker of platelet clumps count was used to differentiate the two diseases for samples anficoagulated with EDTA or citric acid. Results In 65 patients with thrombocytopenia, platelet counts were (48±11)×109/L detected by automatic hematology analyzers. Fifty of 65 cases were true thrombocytopenia which showed low platelet counts [(48±10)×109/L by automated analyzer and (46±11)×109/L by manual assay]. No significance was observed between them (t=-1.26, P0.05). Platelet clumps counts were 86±15. No platelet clamps were detected under microscope. The other 15 cases were EDTA-PTCP [platelet counts were (48±12)×109/L and platelet clumps counts (840±184) were increased significantly by automated analyzer and using EDTA anticoagulant] which showed obviously platelet clumps and no less platelet counts under microscope. After replacing citric acid, platelet counts [(141±13)×109/L by automated analyzer and (134±17)×109/L by manual microscope assay] were increased significantly. No significance was observed between them (t=-1.29, P0.05). Platelet clumps counts (75±12) were decreased obviously compared with EDTA anticoagulant method (t=-6.82, P<0.001). No platelet clumps were detected under microscope. Conclusion Platelet clumps counts may be a useful clinical indicator for monitoring of platelet aggregates, especially for EDTA-PTCP caused by platelet clumping.

Background and purpose:RASSF10 acts as a kind of tumor suppressor in various tumor tissues, but researches in cardiac adenocarcinoma has not been reported. This study aimed to detect the methylation status and expression ofRas-association domain family 10 (RASSF10) in gastric cardia adenocarcinoma (GCA), and explore its role in occurrence and development of GCA.Methods:Methylation speciifc polymerase chain reaction (MSP), reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry method were respectively used to detect methylation status, mRNA expression and protein expression ofRASSF10 in 81 GCA tissues and corresponding normal tissues.Results:The promoter methylation frequency ofRASSF10 in GCA tissues (64.20%, 52/81) was signiifcantly higher than that in corresponding normal tissues (20.99%, 17/81,P<0.05), and was closely correlated with TNM stages, differential degree and lymph node metastasis (P<0.05). RASSF10 mRNA expression in GCA tissues (0.57±0.05) was significantly lower than that in corresponding normal tissues (0.78±0.02,P<0.05), and was closely correlated with TNM stages and lymph node metastasis (P<0.05). Protein expression of RASSF10 in GCA tissues (31.10%, 26/81) was signiifcantly lower than that in corresponding normal tissues (71.60%, 58/81,P<0.05), and was closely correlated with TNM stages, differential degree and lymph node metastasis (P<0.05). The promoter methylation frequency ofRASSF10 in GCA tissues was inversely related to its protein expression.Conclusion:Inactivation of RASSF10 caused by aberrantmethylation in the promoter region may be closely correlated with the GCA tumorgenesis.

Objective To detect the expression of SALL4 in patients with acute myeloid leukemia (AML) and analyze its potential clinical significance. Methods Reverse transcription polymerase chain reaction and Real-time fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) was used to examine SALLA expression in peripheral blood mononuclear cells (PBMCs) of 68 cases of AML including 36 cases in acute phase and 32 cases in remission phase, 30 healthy controls, Kasumi-1 cells and THP-1 cells. Then, flow cytometry, bone marrow smear and automated hematology analyzer were used to analyze the relationship between the SALL4 expression and blast cell counts in the bone marrow, peripheral white blood cell (WBC) counts, peripheral large unstained cell (LUC), CD34 in blast cells. Further, the change of SALL4 level during pre-chemotherapy, chemotherapy (2nd w to 3rd w) and remission were investigated in 5 AML cases. Results The level of SALL4 expression in patients with AML in acute phase [69.01 (17.20-120.28)] was 26-fold and 61-fold high compared with that in remission phase [2.64(1.35-5.41)] and in healthy control [1.14(0.50-1.62)] (Z=-6.48,-6.83,P<0.01). The level of SALL4 expression in remission phase was 2.3-fold high compared with that in healthy control (Z=-3.61 ,P<0.01). The expression level of SALL4 was decreased along with efficient chemotherapy in 5 AML cases in which SALL4 expression level was 79.74 (33.76-89.09), 7.19 (5.97-20.21) and 3.40 (1.44-15.53) during pre-chemotherapy, chemotherapy (2nd w to 3rd w) and remission, respectively. In groups of abnormal increased counts of blast cell, peripheral LUC% and CD34%, expression of SALL4 [33.82 (16.00-144.01), 30.70(23.75-72.50) and 56.25(23.79-153.81), respectively] were higher than that in groups of normal counts [2.74 (1.59-5.13), 5.71 (2.52-22.40) and 20.82 (14.03-55.12), respectively ] (Z=-4.64,-2.18,-3.66,P<0.01 or P<0.05). The expression of SALL4 in the group of increased WBC counts [89.26(23.75-154.34)] was higher than that in the group of normal WBC counts [3.86(2.03-6.01)] and the group of decreased WBC counts [6.66(2.51-17.06)] (Z=-4.91,-4.21,P<0.01). The level of SALL4 expression was positively correlated with blast cell counts in bone marrow and peripheral WBC counts (r=0.45,0.40,P<0.01). Conclusions FQ-RT-PCR method can be used successfully to detect the expression of SALL4,and the expression of SALLA may be useful to predict disease progression of AML.

A moderately halophilic bacterium strain DF-B6 capable of degrading Tween 20 was isolated from solar saltern brine by plate method.Based on the phenotype,physiological and biochemical character-istics,and the phylogenetic analysis of 16S rDNA sequence,strain DF-B6 was identified as Idiomarina.The substrate specificity test showed that the lipolytic enzyme from strain DF-B6 was an esterase,and not a li-pase.The maximum esterase activity was observed in 8% NaCl concentration at 50?C,pH 8.0.The effect of various divalent cations was studied on the activity of esterase from Idiomarina sp.DF-B6 at the concentra-tion of 10 mmol/L,Mg2+,Ca2+ and Mn2+enhanced the esterase activity,while Zn2+,Fe3+ and Cu2+ inhibited its activity.

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.