Objective To analyse the problems presented in external quality assessment (EQA)for blood cytomorphology and propose measures for improvement.Methods Statistical analysis was performed on EQA results for blood cytomorphology from 2011 to 2013 in Shaanxi Province,including general information of participant laboratories,coincident rate,and reported incorrect results.EQA for blood cytomorphology was held two times every year in Shaanxi province,with ten pictures inclu-ding bone marrow and peripheral blood smear each time.The compact disc including twenty pictures was given to partici-pants by EMS.Participants reported two EQA results in April and September each year.The center statisticed the EQA re-sults and provided the EQA reports to every participants in June and November.Results Participating laboratories increased from 76 in 2011 to 163 in 2013.The ratio of laboratories with the coincidence rate≥80% was 80%,47%,44%,55%,77%and 96% respectively.The number for single cell with the coincidence rate≥80% was 38.The coincidence rate of peripheral blood cells was higher than that of the bone marrow on the whole.Causes of incorrect results included cell lines’misclassifi-cation,growth stage’s misclassification,insufficient identification of abnormal cytomorphology,and so on.Conclusion The identification of blood cytomorphology was unbalanced in different leveled hospitals in Shaanxi Province.To develop EQA of blood cytomorphology definitely has a positive role in improving the experimenters’skill of identifying cytomorphology.

ObjectiveLoss-of-function mutation of p53,a tumor suppressor gene,is an important mechanism for the development of human cancers.In this study we tried to transfect p53N15-based fusion peptide into H1299,a lung cancer cell line,and evaluate the anti-tumor effects of the fusion peptide.MethodsAdeno-associated virus ( AAV) vectors were used for transfecting p53N15 fusion peptide into p53-null lung adenocarcinoma H1299 cells.The anti-replication effects of p53N15 fusion peptide were evaluated with inverted microscopy,MTT test for cell viability and flow cytometry.ResultsFusion peptides in H1299 cells was highly expressed and had detectable suppressive effects on cell proliferation.A large amount of dead cells were seen under microscope after the transfection of recombinant viruses for 72 hours.Cells activity was reduced significantly in the virus-transfecting groups as demonstrated by MTT test.The flow cytometry showed that a large number of dead cells were present in the virus-transfecting groups.ConclusionThe growth of H1299 lung adenocarcinoma cells could be inhibited in vitro by being transfected with p53N15 fusion peptide,which may be a potential gene therapy alone or as an adjuvant option in the treatment of lung cancer.

Objective To induce human mononuclear cell of cord blood into CD 3 activating killing (CD 3AK) cells with anti-CD 3 monoclonal antibody (CD 3McAb) and recombinant human interleukin-2 (rhIL-2), so that their proliferative activity, activity of killing action, phenotypes and level of secretory cytokines can be observed dynamically. Methods The increase of the number of cells was counted by Tapan-blue staining. The killing action can be measured by using methyl -thiazolyl-tetrazolium-array. The phenotypes of cells were analysed by using indirect immunofluorescence assay. The levels of IL-6, interferon-? (IFN-?) and tumor necrotic factor-? (TNF-?)in culture supernatants were analysed by using enzyme-linked irnmunosorbent assay(ELISA). Results The increase of the number of CD 3AK cells from cord blood was the highest amounting to 78.56 times in the second week. The killing action reached the peak on day 12, and all target cells (malignant cell lines) could be killed significantly. The heterogeneous phenotypes of CD 3AK cells showed that the number of cells with CD 3+, CD 8+, CD 25+, CD 38+, CD 16+ and CD 56+ increased significantly on day 7,14 compared with those of pre-culture (P

Objective The purpose of this paper is to understand the advantages and disadvantages of the bone marrow smears and bone marrow biopsy in multiple myeloma diagnosis and efficacy judgment,explicit the value of bone marrow smears and bone marrow biopsy synchronous check in the diagnosis and treatment observation of multiple myeloma.Methods With two step-suction two biopsy specimens assay,obtained specimens of bone marrow smears and bone marrow biopsy,retrospective-ly analysed results of 283 multiple myeloma patients bone marrow smear and biopsy,and made a comparative study on the degree of bone marrow hyperplasia,myeloma cell morphology,the degree of tumor cell infiltration,proliferation pattern,bone marrow stromal pathological changes,and fibrosis cases.Results The degree of proliferation of bone marrow biopsy sections and infiltration of plasma cells was significantly higher than that of bone marrow smears,statistically there was a significant difference (P <0.01).Multiple myeloma diagnostic sensitivity by bone marrow biopsy sections was significantly higher than by the bone marrow smears,the difference was statistically significant (P < 0.05).Plasma cells in bone marrow biopsy tumor proliferation modes:clusterpiece nodular type 33 cases (11.66%),interstitial-type 86 cases (30.39%),among nodular interstitial type 112 cases (39.58%),diffuse cypriot real 52 cases (18.37%).Plasma cells in bone marrow smears tumor morphology:small mature plasma cell type 77 cases (27.21%),immature plasma cell type 148 cases (52.30%),protoplas-mic cell type 36 cases (12.72%),reticular plasma cell type 22 cases (7.77%).Conclusion Marrow biopsy can accurately reflect the degree of bone marrow hyperplasia,plasma cell tumor proliferation mode and infiltration degree,myelofibrosis sit-uation;bone marrow smears Wright-Giemsa staining,plasma cell tumor morphology was clear,typicalfeatured,and easily i-dentifiable.Bone marrow smear and biopsy synchronous check can improve the sensitivity and accuracy for multiple myeloma diagnosis,which has very important significance for multiple myeloma diagnosis and treatment observation.

Objective To observe on the clinical effect and the influence of the level of plasma vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR) and basic fibroblast growth factor (bFGF) in acute leukemia before and after treatment by thalidomide combined with chemotherapy. Methods Thirty-six cases of acute leukemia patients were randomly divided into experimental group and control group by 18 cases each. Each group was treated with conventional chemotherapy in the standard-dose, meanwhile in the experimental group additional thalidomide 100 mg/day were taken orally. Before treatment and 8 weeks after treatment, plasma were collected for the detection of VEGF, VEGFR and bFGF content by double antibody sandwich enzyme-linked immunosorbent assay (ELISA).Results The ratio of experimental group and control group, were 88.9 % (16/18), 77.8 % (14/18)respectively and the difference was statistically significant (x2 =4.103, P ＜0.05). The level of plasma VEGF (389.78+249.94 pg/ml, 318.54±125.78 pg/ml) of experimental group and control group before treatment was statistically significant (t = 3.141, t =3.024, P ＜0.01) compared with healthy group [(132.91±26.66) pg/ml] respectively. The level of plasma VEGF of those groups after treatment [(211.74+36.72) pg/ml, (288.02±31.77) pg/ml] was statistically significant (t =2.413, t =2.324, P ＜0.05) compared with healthy group respectively. The difference of the level of plasma VEGF of experimental group and control group before treatment was not statistically significant (t =1.384, P ＞0.05). The difference of the level of plasma VEGF of experimental group and control group after treatment was statistically significant(t =2.793,P ＜0.05). The level of plasma VEGFR [(2490.75+1695.9) pg/ml, (2322.78+1105.87) pg/ml] of experimental group and control group before treatment was statistically significant (t =2.914, t =2.783, P ＜0.01) compared with healthy group [(1134.98+378.45) pg/ml] respectively. The level of plasma VEGFR of those groups after treatment [(1359.71± 390.24) pg/ml, (1753.89±337.04) pg/ml] was statistically significant(t =2.572, t =2.447, P ＜0.05) compared with healthy group respectively. The difference of the level of plasma VEGFR of experimental group and control group before treatment was not statistically significant (t =1.276, P ＞0.05). The difference of the level of plasma VEGFR of experimental group and control group after treatment was statistically significant (t = 2.486, P ＜0.05). The level of plasma bFGF [(2.43±0.27) ng/ml, (2.41±0.33) ng/ml] of experimental group and control group before treatment was statistically significant(t =4.982, t =4.171, P ＜0.05) compared with healthy group (1.83±0.44) ng/ml respectively; the level of plasma bFGF of those groups after treatment [(2.09±0.17) ng/ml,(2.11±0.31) ng/ml] was statistically significant (t =3.011, t =2.773, P ＜0.05) compared with healthy group respectively. The difference of the level of plasma bFGF of experimental group and control group before treatment was not statistically significant (t =0.953, P ＞0.05). The difference of the level of plasma bFGF of experimental group and control group after treatment was not statistically significant (t =1.282, P ＞0.05).Conclusion The remission rate could be improved by thalidomide combined with chemotherapy in acute leukemia, which could be an effective treatment by anti-angiogenesis and inhibiting the growth and infiltration of acute leukemia cells.

Objective To explore the differentiated diagnostic value of the morphological changes of follicular dendritic cell (FDC)meshwork between different subtype of lymphoma.Methods CD21 was stained by immunohistochemistry (IHC) method,FDC meshwork pattern was studied in 5 6 cases of various lymphomas(including 8 cases of diffuse large B cell lym-phoma,2 cases of burkitts lymphoma,6 cases of small lymphocytic lymphoma,6 cases of plasmacytoma,3 cases of MALT lymphoma,6 cases of peripheral T cell lymphoma,3 cases of anaplastic large cell lymphoma,8 cases of NK/T cell lympho-ma,4 cases of follicular lymphoma,3 cases of mantle cell lymphoma,3 cases of AITL,2 cases of FDC sarcoma).Results FDC meshwork in the morphological changes of various subtypes of lymphoma could be classified into four patterns:①FDC form a disappeared and disintegrated meshwork,most of the lymphoma FDC meshwork fully or partially destroyed,including diffuse large B cell lymphoma,burkitt lymphoma,small lymphocytic lymphoma,plasmacytoma,peripheral T cell lymphoma, anaplastic large cell lymphoma,NK/T cell lymphoma;②FDC meshtwork existence,even hyperplasia,including follicular lymphoma,mantle cell lymphoma,MALT lymphoma;③FDC meshtwork proliferation,disorder and deformation,such as AI-TL;④Full expression subtype:such as FDC sarcoma.Conclusion The morphologic pattern of the FDC meshwork in lym-phomas of follicular origin was differs according to the lymphoma subtypes,and it has important clinical value in lymphoma differential diagnostic.

Objective To explore the abnormal karyotype characteristics of myelodysplastic syndrome (MDS)patients and their correlation with clinical prognosis.Methods Analyzed the karyotypes of 281 MDS patients by use of G-banding tech-nique.Results Through analysis of the karyotypes of 281 MDS patients,found that the percentage of abnormal karyotypes was 48.75% (137/281),among 137 patients with abnormal karyotypes,43.07% (59 cases)presented with numerical aber-ration,31.39% (43 cases)with structural aberration,and 25.54% (35 cases)with both numerical and structural abnormali-ties.As for MDS subtypes,the occurrence rate of abnormal karyotype was 63.41% (26/41)in RAEB-2,58.73% (37/63)in RAEB-1,39.2% (49/125)in RCMD,15.38% (2/13)in RAS and 22.58% (7/31)in RA.The rates of abnormal karyotype in RAEB-1 and RAEB-2 were significantly higher than that in RA and RAS(P<0.01),and in RCMD (P <0.05).The fre-quent abnormal karyotypes were as follows:+8,-7/7q-,-20/20q-,complex karyotypes chromosomal translocation,i(17),-Y and +21.The follow-up study of 159 MDS patients indicated that the median survival time was 39 months for 68 patients with normal karyotypes and 21 months for 91 patients with abnormal karyotypes,the former was significantly prolonged than the latter (P < 0.05).As far as the leukemia transition rate was concerned,the patients with aberrant karyotypes (35.5%)were significantly higher than that with normal karyotypes (10.3%)(P < 0.01),among them,the cases with complex karyotypes and-7/7q-more easily transit into leukemia.Conclusion MDS was one kind of clonal hematological ma-lignancy with high heterogeneity.Chromosomal karyotype test plays an important role in the correct diagnosis,typing and prognosis evaluation of MDS.

OBJECTIVETo establish a serum-free culture system of dendritic cells (DCs) from chronic myeloid leukemia (CML) cells so that DCs vaccine may be applied to the adoptive immunotherapy of CML in the near future.

METHODSFetal calf serum, serum-free medium and autologous serum were used for culture of DCs. The usage of cytokines was classified into two groups: group A (stem cell factor, granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4) and group B (granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4). The phenotypes of DCs were analyzed by using indirect immunofluorescence and flow cytometry. Mixed leukocyte responses were performed by methyl thiazolyl tetrazolium (MTT) assay. Chromosome analysis of DCs can be achieved by displaying G banding. T cells from CML patients were stimulated with autologous DCs and T-cell cytotoxicity was measured by (MTT) assay.

RESULTSCD34(+) cells or mononuclear cells were obtained from peripheral blood or bone marrow samples of eight patients of chronic-phase CML. Group A of serum-free medium was better than group B in expansion of total cell numbers and the rate of DCs. These results of serum-free medium were not significantly different from those of fetal calf serum medium, but the results of autologous serum medium were inferior to two groups above. The expression of major histocompatibility complex class II antigen on the surface of DCs was notable (> 50%), but the expression of CD83 and the costimulatory molecules CD86 was not noticeable (10% - 50%). Although CD1a(+)/CD14(-) DCs were potent stimulators of allogeneic lymphocytes, expansion of T cells from normal volunteers were not significant (average 27.2 fold at DCs: T cells ratio of 1:10). At day 12, CD1a(+) cells from three patients were studied by displaying G banding and Ph(+) cells in these populations were 100%, 98% and 60%, respectively. At an effector: target ratio of 40:1, 32% to 45% cytotoxicity was noted with DC-stimulated T cells against autologous leukemia cells.

CONCLUSIONSA stable serum-free culture system of CML-DCs was established. The expression of CD83 and CD86 on the surface of CML-DCs and DCs' potent stimulation of allogeneic lymphocytes were not notable. DCs in CML patients can be derived from the malignant clone and these malignant DCs could induce anti-leukemic reactivity in autologous T lymphocytes without the necessity for additional exogenous antigens.

Cells, Cultured ; Culture Media, Serum-Free ; Cytotoxicity, Immunologic ; Dendritic Cells ; physiology ; Humans ; Immunotherapy, Adoptive ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; therapy ; T-Lymphocytes ; immunology

Objective: To observe the analgesic and anti-inflammatory effect of mandelic acid. Methods: Fifty Kunming mice were randomly divided into 5 groups:the blank control group (0. 1 ml/10 g), mandelic acid high (300 mg·kg-1), medium (200 mg ·kg-1 ) and low (140 mg·kg-1 ) dose groups, and the positive control ( aspirin) group, ig, qd. The analgesic effect of mandelic acid was observed by writhing test and hot plate method in mice. The ear swelling model caused by dimethyl benzene in mice was a-dopted to observe the analgesic effect. Results:Mandelic acid in each dose group could make the number of writhing in mice signifi-cantly reduced and pain threshold extended, and when compared with the blank control group, the difference was statistically significant (P<0. 01). The writhing times of mice mandelic acid high dose group was fewer than that of the positive control group, and there was no statistically significant between the groups (P>0. 05). In low and medium dose group, the writhing times of mice were more than those of the positive control group, and there was a significant difference between the low dose group and the positive control group( P<0. 05). The pain threshold of the mice in each mandelic acid dose group was higher than that of the positive control group, the pain threshold increased significantly in the high dose group before and after the administration, and the difference was statistically signifi-cant when compared with the positive control group (P<0. 05). The effect of mandelic acid on the ear swelling of mice was not signifi-cant, and when compared with the blank control group, the difference was not significant (P>0. 05). Conclusion:Mandelic acid has significant analgesic effect, while anti-inflammatory effect is not obvious.

【Objective】 To analyze the testing results of platelet antibody in peripheral blood of 266 pregnant women, so as to explore the correlation of platelet antibody between adverse pregnancy and the number of pregnancies. 【Methods】 A total of 266 pregnant women, admitted to Shaanxi Provincial People′s Hospital, were selected for platelet antibody detection. They were divided into two groups according to history of adverse pregnancy, and the positive rate of platelet antibody between two groups was compared. They were divided into group A (1 pregnancy), group B (2 pregnancies) and group C (≥3 pregnancies) in parallel, and the positive rate of platelet antibody among three groups was compared 【Results】 The yield rate of platelet antibody in groups with and without adverse pregnancy was 31.81% vs 14.86%, showing statistical significance(P<0.05). The yield rate of platelet antibody of Group A, B and C was 9.09%, 21.62% and 23.65%, respectively, with The significant differences(P<0.05). The statistical analysis of inter-group χ² test demonstrated a linear trend between the number of pregnancies and platelet antibodies yielding(χ²=7.056, P<0.05). 【Conclusion】 Platelet antibody was correlated with adverse pregnancy, and the yield rate of platelet antibody in the group with adverse pregnancy was higher than that in the group without adverse pregnancy. There was a linear trend between platelet antibodies yielding and the number of pregnancies. The yield rate of antibodies gets higher as the number of pregnancies increased. Taking platelet antibody test as a routine test during pregnancy is conducive to the early detection of clinical adverse pregnancy, as well as the early prevention, early detection and early intervention of fetal/neonatal homoimmune thrombocytopenic purpura and other diseases.