Objective To observe the general status and the immunity of DBA/2 mice injected with leukemia vaccine during the immune reconstitution, and to observe the survival rate of these injected mice that were then challenged by L1210 cells. Methods Normal mice and reconstituted lymphopenic mice (RLM; DBA/2 mice rendered lymphopenic with sublethal irradiation and reconstituted with naive splenocytes) were used in the vaccination and challenge experiments with L1210 cells. Results Following vaccination, a significant increase of the IFN-? was detected in the blood of vaccinated RLM compared with that of vaccinated normal mice, as assessed by ELISA. Significantly, greater protection was induced in vaccinated RLM, which led to a stronger protection from death. Conclusion The vaccination of reconstituted lymphopenic hosts could elicit superior anti-tumor immunity compared with normal hosts, highlighting the potential clinical benefit of performing tumor vaccination during immune reconstitution.

Objective To explore the capacity of the Sweden CellaVision DM96 automatic digital cell morphology analysis sys‐tem (DM96) in nucleated cell classification of serous cavity effusion .Methods 36 specimens of serous cavity effusion were selected from the inpatients of Second Affiliated Hospital of Xi′an Jiaotong University in March 2015 and performed the Wright staining by the two kinds of method ,the Japanese Sysmex SP‐1000Ⅰ automatic smearing machine and manual smearing ,after staining ,the smear was performed the nucleated cells classification by DM96 .The consistency and relevance of the classification results by DM 96 with those by the Sysmex XT‐4000i were calculated .Results The classification results by DM96 had better consistency with the results by XT‐4000i ,moreover the cell images taking by DM96 were clear with high automatic degree .Conclusion The DM96 auto‐mated digital nucleated cell morphology analysis system is reliable and effective ,and has a significance for improving the cellular morphological analysis of serous cavity effusion specimen .

The frequency of peripheral lymphocyte subsets using monoclonal was studied in patients with viral myocarditis (MC). The percentage of T_8 positive cells was significantly lower in patients with MC of heart failure(MCHF) than that in patients with MC of arrhythmias and that in normal controls, whereas the T_4/T_8 ratio was significantly inereased in MCHF patients. At the same time, suppressive T cells (Ts) function in patients with MC was evaluated by shortlived assay. The Ts function was significantly reduced in MCHF patients, close correlation was found between the Ts function and the hemodynamic function in MCHF patients. Abnormal immunoreguiation may be of the important pathogenic factor in MCHF patients.

Our previous study demonstrated that human KIAA0100 gene is a novel acute monocytic leukemia-associated antigen (MLAA) gene. But the functional characterization of human KIAA0100 gene has remained unknown to date. Here, firstly, bioinformatic prediction of human KIAA0100 gene was carried out using online software;Secondly, human KIAA0100 gene expression was downregulated by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system in U937 cells. Cell proliferation and apoptosis were next evaluated in KIAA0100-knockdown U937 cells. The bioinformatic prediction showed that human KIAA0100 gene was located on 17q11.2, and human KIAA0100 protein was located in the secretory pathway. Besides, human KIAA0100 protein contained a signal peptide, a transmembrane region, three types of secondary structures (alpha helix, extended strand, and random coil) , and four domains from mitochondrial protein 27 (FMP27). The observation on functional characterization of human KIAA0100 gene revealed that its downregulation inhibited cell proliferation, and promoted cell apoptosis in U937 cells. To summarize, these results suggest human KIAA0100 gene possibly comes within mitochondrial genome; moreover, it is a novel anti-apoptotic factor related to carcinogenesis or progression in acute monocytic leukemia, and may be a potential target for immunotherapy against acute monocytic leukemia.

Monoclonal antibodies (MAbs) are important tools for the study of proteins′ function and structure. But there has been no report on the preparation of MAbs against human KIAA0100 protein up to date. Here, first, we generated the mouse MAb against human KIAA0100 protein using purified recombinant 6×Histidinc (6×His)- tagged human KIAA0100 protein segment (1557–2234) as an antigen; then, the mRNA expression of human KIAA0100 gene was detected in U937 cells using Northern blot analysis. The results showed that the mouse MAb against human KIAA0100 protein could sensitively recognize the human KIAA0100 protein using Western blot analysis and immunocytochemistry analysis. Besides, Western blot analysis revealed that human KIAA0100 gene possibly encoded two different protein products (254 kDa and < 250 kDa) in U937 cells. Moreover, Northern blot analysis confirmed that human KIAA0100 gene might produced two different mRNA products (6000–10000 bp and 5000–6000 bp) in U937 cells. The results provide a basis for large-scale production of the MAb against human KIAA0100 protein, which will be useful for the study of human KIAA0100 protein′s function/structure and MAb-targeted drugs in the future.

Objective To investigate the relationship of acute graft versus host disease (aGVHD) and the regulatory T (Treg) cells,NK cells as well as their function-related cytokines such as IL-10,TGF-β,and perforin in mice with allogeneic bone marrow transplantation.Methods H-2 completely mismatched C57BL/6→ BALB/c aGVHD mice model was constructed.Flow cytometry analysis was used to detect the proportion of CD4+CD25 + Treg cells and NK-1.1+ NK cells in the spleen of aGVHD mice after transplantation.ELISA method was used to detect the serum levels of IL-10,TGF-β and perforin in the aGVHD mice intervened with CSA prophylactic or not.The normal C57BL/6 mice were used as controls.Results Compared with those of normal mice,the proportion of Treg cells in aGVHD mice after transplantation was decreased (mean 3.6％ vs.1.55％) and the proportion of NK cells increased (mean 3.3％ vs.11.5％).In the aGVHD mice treated with cyclosporine A,serum IL-10 expression level was significantly increased (treated group (125.79 ± 0.27)pg/mL,untreated group [(103.09 ± 3.27)pg/mL,P＜0.01)],TGF-β expression level was increased [(252.05 ±7.84)pg/mL vs.(241.61±15.41)pg/mL,P＞0.05],perforin expression level was significantly increased [(186.97 ± 4.68)pg/mL vs.(144.35 ± 14.42)pg/mL,P＜0.01].Conclusion ① The occurrence of aGVHD is correlated with the decreased number of Tree cells after transplantation in mice.Treg cell function-related cytokines IL-10 and TGF-β are involved in the treatment of aGVHD by cyclosporine A-mediated immunosuppression.②NK cells are involved in the occurrence of aGVHD after allogeneic bone marrow transplantation,and the increased level of perforin is related to the inhibition of aGVHD.

Objective:To observe the general status, the effect of resisting to the aggression of FBL-3 cells and protective immunity of the mice by using leukemia vaccine during the immune reconstitution.Methods:The immune deficiency C57BL/6 mice induced by total body irradiation were reconstituted immunity with homologous splenic lympocytes.The mice were immunized by injecting FBL-3 cells subcutaneously,and then attacked by using FBL-3 cells 7days thereafter.The activities of mice,the percentage of carcinoma formation and the growth status of tumor were observed.The amount of IFN-? in peripheral blood was determined by ELISA to evaluate the magnitude of cell immunity.The condition of cell infiltration in tumor central region and its periphery region was explored by patologic-section.Results:Immune reconstitution could obviously elevate protective immunity on the basis of using vaccine,and the amount of IFN-? was obviously higher.The FBL-3 tumors inoculated thereafter grwe slowly,and significant infiltration of mononuclear cells peri-tumor as well as necrosis of tumor cells were observed.Also,the life time of the mice was prolonged.Conclusion:Immune reconstitution and vaccine can significantly elevate protective immunity

Objective To study the curative effects and adverse effects of the thalidomide combined with COMP chemotherapy in treating multiple myeloma(MM).Methods 42 patients were initially diagnosed as MM and 27 patients were refractory and relapsed MM.The small dose of thalidomide combined with COMP management and COMP management alone were used.The effective rate and adverse effects were analyzed.Changes of M-protein in serum,percentage of plasma cells in bone marrow and the level of hemoglobin were also analyzed in both pre-treatment and post-treatment periods.Results In 42 patients who were initially diagnosed as MM,the effective rate was 40.9% for 22 patients treated by chemotherapy alone and 70.0% for 20 patients treated by the thalidomide combined with chemotherapy.Statistic difference was observed between those two group.As to the 27 patients who were refractory and relapsed MM,the effective rate was 42.9% for 13 patients treated by chemotherapy alone and 84.6% for 14 patients treated by the thalidomide combined with chemotherapy.Statistic significance was present between them.Adverse effects were less and tolerated.Conclusion Treatment of small dose of thalidomide combined with COMP chemotherapy could significantly improve the effective rate with less adverse effects.

Objective To construct a cDNA expression library of Psilgramma menephorn to screen its major allergen so as to provide the basis for producing recombinant allergen vaccine of Psilgramma menephorn. Methods Total RNA was extracted from the whole body of Psilgramma menephorn with Trizol and mRNA was purified with Oligo (dT) Spin-Column. And dscDNA was synthesized through reverse transcription. After blunting, the cDNA fragments were ligated with EcoRⅠ adapters. Then the cDNAs were digested by XhoⅠ, and the fragments less than 400 bp were removed by using GHROMA SPIN-400 column. The remaining fragments longer than 400 bp were ligated with Uni-ZAP XR vector. The recombinants were packaged in vitro and a small portion of the packaged phage was used to infect E.coli XL1-Blue MRF′ for titration. The recombinants were examined by color selection. The size of cDNA inserts and the diversity of library were analyzed by PCR. The library was screened using SPT positive sera from patients with Psilgramma menephorn allergy repeatedly. Results The cDNA expression library consisting of a 5×105 recombinant bacteriophages was constructed with the recombinant ratio of 67%. The average length of recombinant exogenous inserts was about 1.49 kb. Five positive cDNA clones were obtained. Conclusion The constructed cDNA expression library shows appropriate contents and size of cDNA fragments and the related genes of Psilgramma menephorn major allergens were harbored successfully, which lays the foundation for the positive clone identification and further analysis.

Objective:To evaluate the cytotoxicity of macrophage of C57BL/6 mice treated with the prepared leukemia vaccine.Methods:Established the leukemia burden mice and prepared three types of leukemia vaccine,which were administered on the mice respectively.The MTT colormetric method was adopted 2 and 4 weeks later to measure the cytotxicity of M? derived from the mice accept immunotherapy or prevention,and compared with control group.Results:(1)With the growth of leukemia cells in the mice,the cytotoxicity of M? was seriously depressed;(2)The administration of tumor vaccine prepared from inactivated tumor cells,IFA and cytokines,such as rGM CSF,rIL 2 and rIL 6,promote the cellular immunity of tumor burden mice,more effciently than tumor vaccine made from either inactivated tumor cells and IFA or inactive tumor cells themselves along.Conclusion:The tumor vaccine simply prepared with inactive tumor cells,IFA and cytokines can active the non specific cellular immunity that represented as M?,for example non specific cytotoxicity,antigen present,immune surveillance and so on,these can build a base for active the T lymphocyte the next.The tumor vaccine provides a promising future in the therapy against hematopietic tumor.