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Objective:To evaluate the performance of an artificial intelligent (AI)-based automated digital cell morphology analyzer (hereinafter referred as AI morphology analyzer) in detecting peripheral white blood cells (WBCs).Methods:A multi-center study. 1. A total of 3010 venous blood samples were collected from 11 tertiary hospitals nationwide, and 14 types of WBCs were analyzed with the AI morphology analyzers. The pre-classification results were compared with the post-classification results reviewed by senior morphological experts in evaluate the accuracy, sensitivity, specificity, and agreement of the AI morphology analyzers on the WBC pre-classification. 2. 400 blood samples (no less than 50% of the samples with abnormal WBCs after pre-classification and manual review) were selected from 3 010 samples, and the morphologists conducted manual microscopic examinations to differentiate different types of WBCs. The correlation between the post-classification and the manual microscopic examination results was analyzed. 3. Blood samples of patients diagnosed with lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndrome, or myeloproliferative neoplasms were selected from the 3 010 blood samples. The performance of the AI morphology analyzers in these five hematological malignancies was evaluated by comparing the pre-classification and post-classification results. Cohen′s kappa test was used to analyze the consistency of WBC pre-classification and expert audit results, and Passing-Bablock regression analysis was used for comparison test, and accuracy, sensitivity, specificity, and agreement were calculated according to the formula.Results:1. AI morphology analyzers can pre-classify 14 types of WBCs and nucleated red blood cells. Compared with the post-classification results reviewed by senior morphological experts, the pre-classification accuracy of total WBCs reached 97.97%, of which the pre-classification accuracies of normal WBCs and abnormal WBCs were more than 96% and 87%, respectively. 2. The post-classification results reviewed by senior morphological experts correlated well with the manual differential results for all types of WBCs and nucleated red blood cells (neutrophils, lymphocytes, monocytes, eosinophils, basophils, immature granulocytes, blast cells, nucleated erythrocytes and malignant cells r>0.90 respectively, reactive lymphocytes r=0.85). With reference, the positive smear of abnormal cell types defined by The International Consensus Group for Hematology, the AI morphology analyzer has the similar screening ability for abnormal WBC samples as the manual microscopic examination. 3. For the blood samples with malignant hematologic diseases, the AI morphology analyzers showed accuracies higher than 84% on blast cells pre-classification, and the sensitivities were higher than 94%. In acute myeloid leukemia, the sensitivity of abnormal promyelocytes pre-classification exceeded 95%. Conclusion:The AI morphology analyzer showed high pre-classification accuracies and sensitivities on all types of leukocytes in peripheral blood when comparing with the post-classification results reviewed by experts. The post-classification results also showed a good correlation with the manual differential results. The AI morphology analyzer provides an efficient adjunctive white blood cell detection method for screening malignant hematological diseases.
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Objective This study is aimed to investigate the prognostic significance of ring sideroblast ( RS) in MDS( Myelodysplastic Sydrome ) and evaluate the correlation of RS and other prognostic index.Methods A total of 198 patients with MDS between March 2009 and December 2015 in Chinese PLA′s Gerneral hospital were chosen for this study .Based on the ratio of RS in nucleated red blood cell , patients were first separated into myelodysplastic syndrome without ring sideroblast (MDS RS-) group, RS≥15%, and myelodysplastic syndrome with ring sideroblast ( MDS RS +) group, RS <15%. Then, according to the proportion of blasts in bone marrow nucleated cells above 5%or below, patients were further divided into myelodysplastic syndrome with low blasts without ring sideroblast ( MDS-LB RS-) group, myelodysplastic syndrome with low blasts and ring sideroblast ( MDS-LB RS+) group, refractory anemia with excess blast without ring sideroblast ( RAEB RS-) group and refractory anemia with excess blast and ring sideroblast ( RAEB RS+) groupe.All patients had completed the morphological , genetics , molecular biology examination at dignosis, and followed up by phone.The results of the overall survival (OS) analysis have been presented in a Kaplan-Meier curve and cox regression model .Last, according to the percentage of RS in nucleated red blood cell , patients were separated into RS <5%groupe, 5%-15%group, 15%-40%group, RS≥40%group, and analyse their survival prognosis by statistical methods .Results Comparing to MDS RS-group, the morbidity age, WBC and PLT count were significantly higher [61 ±1.91 vs 52 ±1.37, t=-3.555, P<0.01, 3.82(0.47-323)vs 2.6(0.6-59.7), z=-4.014, P<0.01;139.5(7-608) vs 60(3-724), z =-3.988, P<0.01], bone marrow eythroid hyperplasia and gigantocyte were more obvious in MDS RS+group[χ2 =11.032, P<0.01, χ2 =5.165, P<0.05]; the percentage of GATA1 gene and abnormal rate of poor prognosis gene ( MLL, NRAS, WT1 ) , either mutation or high gene expression , were higher in MDS-LB RS+group than that in MDS-LB RS-( P<0.05 ); Contrasting with RAEB RS-group, the karyotype is worse in RAEB RS +group[χ2 =4.966, P<0.05];Comparing to 15%-40%group, the OS were poorer in RS≥40%;MDS RS+patients were more prone to adverse prognosis than MDS RS-patients.Conclusion Compared to MDS RS-group, MDS RS +patients had worse prognosis;RS maybe correlate to morbidity age , eythroid dysplasia and gene abnormality in affecting the survival prognosis of MDS.
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The first genetic linkage map of Salvia miltiorrhiza was constructed in 94 F1 individuals from an intraspecific cross by using simple sequence repeat (SSR), sequence-related amplified polymorphism (SRAP) and inter-simple sequence repeat (ISSR) markers. A total of 93 marker loci in the linkage map, consisting of 53 SSR, 38 SRAP and 2 ISSR locus were made up of eight linkage groups, covered a total length of 400.1 cm with an average distance of 4.3 cm per marker. The length of linkage groups varied from 3.3 -132 cm and each of them included 2-23 markers, separately. The result will provide important basis for QTL mapping, map-based cloning and association studies for commercially important traits in S. miltiorrhiza.
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Objective To study the changes of coagulation function and platelet-related parameters of patients with acute pancre-atitis(AP) and their clinical significance .Methods 36 patients with AP were served as the AP group ,38 healthy people ,the control group .Healthy people in the control group and patients in the AP group at the time of admission and remission were subjected to detection of serum amylase ,lipase ,leukocytes ,neutrophils ,lymphocytes ,platelets ,mean platelet volume(MPV) ,platelet distribution width(PDW),prothrombin time(PT),thrombin time(TT),activated partial thromboplastin time(APTT),prothrombin activity (PTA) ,international normalized ratio (INR) ,fibrinogen(FIB) and D-dimer(D-D) levels .Results Levels of serum amylase ,lipase , TT ,INR ,FIB ,and D-D of patients in AP group were significantly higher than those in the control group(P<0 .01) ,while their PT , PTA levels were significantly lower than those in the control group (P<0 .01) .Differences of leukocytes ,neutrophils ,lymphocytes and MPV of AP patients between at the time of admission and remission were statistically significant (P<0 .01) .Conclusion De-tections of coagulation function and platelet-related parameters changes of AP patients contribute to evaluation of the patients′con-dition and prognosis .
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Objective To determine the antigen-specific CTL in PBMC induced by a fusional family-gene vaccine of the immunoglobulin heavy chain variable gene framework region combined with the sequence of cytokine CM-CSF in vitro with MHC pentamer. Methods Peripheral blood samples were collected from two healthy donors and two patients. One was follicular lymphoma and another was hair cell leukemia. PBMC were isolated by density gradient centrifugalization with Ficoll and then subsequently differentiated into immature DCs (imDCs) induced by recombinant human GM-CSF and recombinant human IL-4. Gene gun was used to deliver the plasmids of the gene vaccine or the control plasmids into the imDCs. RT-PCR and ELISA assay were used to detect IgHVl-GM-CSF mRNA and GM-CSF in order to validate the transfection of the vaccine. After adding the cytokine cocktail, the imDCs became mature DCs. Then the mature DCs were co-cultured with lymphocytes from the blood samples for the induction of the antigen-specific CTL. The cultured cells were classified into vaccine group and control group and harvested at different time points of 0 d,7d, 17 d and 24 d after transfection. The subset of CD3+CD8+ T cells was analyzed by FCM assay. Finally, the CTL levels were detected with fluorescently labeled MHC pentamer antibody targeting vaccine epitopes. Results With the induction of cytokines, the imDCs with typical morphology were generated in PBMC. After delivering, the efficient expressions of the vaccine in the imDCs were determined by RT-PCR. And ELISA results also confirmed that GM-CSF was produced at a level of (28 ±6) ng/106 cells of the imDCs loaded with the vaccine, which was significantly different from that of control group (10 ± 3) ng/106 cells (t = 5. 191, P <0.01). FCM assay result showed that the CD3+ CD8+ T cells increased in a stepwise pattern during the culture. For control group, the levels at 0 d,7d, 17d and 24 d were ( 34. 24 ± 2. 72 )% , (46.06 ± 3.08)%, ( 65. 34 ± 4. 26 )% and (73.86 ±4.85 )% , respectively. For vaccine group, the results were (32. 28 ± 2. 08 ) % , (45. 32 ± 3. 81)% , ( 63. 37 ± 4. 21)% and (75. 01 ±3. 20)%. The differences between each time point had statistical significance (F = 176. 966 ,P <0.01) ,but there was no statistical differences between vaccine group and control group ( F = 0.657,P>0.05). The MHC pentamer analysis showed that the DCs loaded with IgHV1-GM-CSF fusional vaccine could efficiently induce the antigen-specific CTL response and the CTL levels increased gradually with the culture time, with the highest level of 4. 36% in the lymphoma blood and 3. 89% in the hair cell leukemia blood. Conclusions MHC pentamer assay could efficiently determine the antigen-specific CTLs response induced by the gene vaccine of family IgHV frame region in vitro. It could be a useful method for monitoring of anti-tumor cell immunity and evaluating of diagnosis and prognosis of the tumors in clinical application.