The present article introduced the standards of moldproof test and their application in detail, expatiated on the harmful effect of the mold on industrial materials or products and the requirements of selecting the experiment parameters, discussed the limitation of the standards for moldproof test used in China.

The direct ELISA was used to detect cells from pure cultures and nodules of Bradyrhizobium japonicum USDA110 and Rhizobium fredii RTt50. The optimum dilution of enzyme-linked conjugates, HRP-Ab110 and HRP-Ab50, was 1:3200 and 1:800 respectively. The optimum dilution of antibodies, Ab110 and Ab50, was 1:3200 and 1:800 separately. The optimum concentrations of antigens, USDA110 and RTt50, were both 6?10~7 cells/ml. Slow and fast-growing soybean rhizobiz can be detected and differentiated specifically by direct ELISA. Among a few strains of these two groups of soybean rhizobia, cross-reaction occurred. This was eliminated by absorption, so that specific strain could be identified by ELISA. The minimal concentration of antigen for detected was 2?10~5 cells/ml. It was found that nodules preserved by drying over silica gel or freezing were equally good, without loss in sensitivity of ELISA. ELISA was used to study the competition of USDA110 and RTt50 with indigenous rhizobia in soil pot experiment. Nodule occupacy of USDA110 ranged from 75-87.5% in different growing season of soybean and RTt50 ranged from 25-45%. The result showed that ELISA was more sensitive than agglutination.

Objective To investigate the effect of mesenchymal stem cell(MSC) on the balance of Th1/ Th2 with different stimulations.Methods To inoculate MSC in 24-well tissue culture plates.after 3 days, added T lymphocytes co-stimulated by PMA and Ionomycin,or the MSC were added to the two-way mixed lymphocyte culture according to different proportion.The subsets and cytokines of T lymphocytes were analyzed by flow cytomety.Results Differentiation into Th1 of T lymphocytes activated by co-stimulation can be inhibited MSC;In MLC,CD8~+T cell subsets and Th1 were evidently decreased.But Th2 cells were slightly increased.Conclusion MSC can significantly suppress CD8~(+)T cell,Th1,increase Th2.MSC has potentialities of alleviating acute graft-versus-host disease(aGVHD) and maintaining graft versus leukemia(GVL).

Objective To investigate the immunophenotype characteristics,the clinical significance of CD28 and International staging system in multiple myeloma (MM).Methods BM aspirate samples from 49 newly diagnosed MM patients and 22 patients after treatment were assessed using 4 color flow cytometric analyses.These MM patients were classified according to International staging system and the consolidated risk staging system,and the survival rates and treatment efficiency of 2 staging system were compared.Results The higher stage,according to the international staging system,the lower treatment efficiency [the efficacy rates of Ⅰ,Ⅱ,Ⅲ stage were 83.3 ％ (4/6),38.5 ％ (5/13),38.5 ％ (5/13),36.5 ％ (19/52)](x2 =4.235,P =0.04),and the survival and progression-free survival time of high-risk group were significantly shortened (P ＜ 0.05).The rate of CD28 abnormal expression was no significant different between the initial and the treatment group [27 (55.1％),14 (63.4 ％)],also similar in the impact of survival and progression-free survival time (P ＞ 0.05).According to the consolidated risk staging system,the treatment efficiency,the survival and progression-free survival time of high-risk group were significantly shortened than the low ones (P =0.040).Conclusions The patients with higher ISS stage,abnormal expression of CD28,and high-risk group have poor prognosis.

Objective To explore the clinical significance of the coagulation and fibrolysis parameters changes for the knowledge of complicated thrombosis after chemotherapy in malignant lymphomas. Methods Morning fasting anti-coagulation blood samples were taken to detect plasma prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and thrombin time (TT) with automatic coagulation analyzer in 71 hospitalized malignant lymphomas and 20 normal controls. The plasma D-dimer levels of the two groups were detected with immunoturbidimetry. Results The levels of plasma APTT, Fib and D-dimer in 71 malignant lynphomas were (30.44±1.43) s, (3.28±0.20) g/L, (297.05±56.59) μg/L respectively, which were significantly higher than those in normal controls at the levels of (23.72±0.76) s, (2.57±0.22) g/L, (94.50±26.07) μg/L respectively (P ＜0.05). The coagulation and fibrolysis parameters were of no statistic differences between the normal controls and lymphomas of stage Ⅰ and Ⅱ (P ＞0.05). The APTT and Fib levels in lymphomas of stage Ⅲ and Ⅳ were higher than those in normal controls and lymphomas of stage Ⅰ and Ⅱ (P ＜0.05). There are 7 malignant lymphomas complicated venous thrombosis . Of the 7 cases, the FIB and D-dimer levels were higher than those of stage Ⅰ and Ⅱ, furthermore the D-dimer levels were higher than those of stage Ⅲ and Ⅳ. Conclusion The abnormalities of coagulation and fibrolysis parameters occurred in malignant lymphomas with requirement of periodic monitoring. After chemotherapy, the lymphoma patients had a high incidence of venous thrombosis, which need early prevention for prolongation of the survival.

Objective: The study was designed to evaluate the changes and significance of circulating CD4~+CD25~+ and CD8~+CD28~- regulatory T cells (Tregs) in patients with multiple myeloma (MM).Methods:CD4~+CD25~+ and CD8~+CD28~-Tregs in peripheral blood of 38 patients with MM and of 20 healthy doners were measured by flow cytometry.Serum albumin and β_2-MG in patients with MM were measured using bromocresol green method,transmission turbidimetry respectively.Results:Compared to those of the controls,the proportions of CD4~+CD25~(+/high),CD4~+CD25~(high) CD127~(low) and CD8~+CD28~-Treg cells in newly diagnosed MM patients were elevated.Furthermore,the proportions of CD4~+CD25~(high) and CD4~+CD25~(high)CD127~(low) Tregs in each clinical stage were elevated when compared to those of the controls.The number of the Tregs were increasing with clinical stages and were significantly higher in stage Ⅲ MM than in stageⅠ MM;In stageⅡand Ⅲ MM,there were also elevated proportions of CD8~+CD28~- Tregs,increasing with clinical stages.However,there were no differences when compared between stage Ⅰ MM and the controls;Both the proportions of CD4~+CD25~(+/high) and CD4~+CD25~(high)CD127~(low) Tregs in active MM were not different from stable MM,although all of them were higher than those of controls.The proportion of CD8~+CD28~- Tregs was higher in active MM than in stable MM and controls,but there were no differences when compared between active and stable MM.The proportions of both CD4~+CD25~(high) Tregs and CD4~+CD25~(high)CD127~(low)Tregs had negative correlation with the levels of serum albumin.Conclusion:MM patients have elevated levels of circulating CD4~+CD25~+ and CD8~+CD28~-Tregs,which may be an important mechanism of MM immune evasion,and may be associated with clinical stages,disease progression and prognosis of MM to some extent.

Objective To compare the efficacy and safety between flumatinib and imatinib in patients with newly diagnosed chronic myeloid leukemia (CML). Methods A multi-center, randomized and parallel comparison clinical trial was conducted in 24 newly diagnosed patients with Philadelphia chromosome-positive CML-chronic phase (Ph+ CML-CP) who were treated by flumatinib 400 mg/d, 600 mg/d or imatinib for 6 cycles (24 weeks). The hematology was evaluated at pre-medication and the 2nd, 4th, 6th, 8th, 10th, 12th, 16th, 20th, 24th week of post-medication. The morphology, cytogenetics and molecular biology were evaluated at pre-medication and 12th, 24th week of post-medication. Results In terms of efficacy, the main molecular remission (MMR) rate of flumatinib 600 mg/d group was higher than that of imatinib group after 24 weeks [44.44 % (4/9) vs. 14.29 % (1/7), P=0.017]. The rate of bcr-ablIS≤10 % in flumatinib 600 mg/d group was significantly higher than that in imatinib group (P=0.002). PK/PD analysis also hinted that patients treated by flumatinib 600 mg/d was more likely to get molecular reaction in the early stage compared with those treated by flumatinib 400 mg/d. In terms of safety, there was no significant difference in grade Ⅲ-Ⅳ of adverse events among flumatinib 400 mg/d group, flumatinib 600 mg/d group and imatinib group (P >0.05). The common adverse events in flumatinib group included skin toxicity, gastrointestinal reactions and diarrhea.There was no heart and cardiovascular toxicity in flumatinib group, and incidence of edema in flumatinib group was lower than that in imatinib group. Conclusions Flumatinib is a safe and effective drug for newly diagnosed patients with Ph+ CML-CP, and 600 mg/d is the appropriate clinical starting dose. Flumatinib and imatinib have similar safety in clinic.

Objective To investigate the common chromosome abnormalities in the patients with multiple myeloma and the relationships of cytogenetic abnormalities and clinical features. Methods The interphase fluorescence in situ hybridization (I-FISH) analysis method was designed to detect RB1-/13q14-and 14q32 rearrangements in 49 MM patients. The statistic value of its effect on clinical features were determined. Results FISH disclosed 14q32 translocations in 26 of the 40 (53.1%) patients. 25 out of the 49 (51.02 %) cases were found with deletion of chromosome 13q14 included del(RB1) in 9 (18.4 %) and del(13q14.3) in 18 (36.7 %). 13q14 deletion and 14q32 translocation were simultaneously observed in 18 (36.7 %) cases. Spearman correlation analysis were found associated of 14q32 rearrangement with the percentage of plasma cells in bone marrow (r=0.316, P=0.27). Conclusion The frequency of 13q14 deletion and 14q32 gene translocation in multiple myeloma are high. There is a significant correlation between the presence of 14q32 translocations and chromosome 13 abnormalities in MM patients. The percentage of 14q32 translocation in plasma cells was increased significantly. The 14q32 translocation is an independent prognostic factor.

Objective To investigate aberrations of bcl-6,p53,c-myc genes in diffuse large B-cell lymphoma (DLBCL) and its clinical significance.Methods Interphase fluorescence in situ hybridization (I-FISH) was detected in 59 DLBCL patients in vivo tissue bcl-6,p53 protein,c-myc gene status.The patients were treated with CHOP or R-CHOP chemotheralpy,and the survival rates and treatment efficiency were compared.Results The p53 deletion was detected in 18 of the 59 cases (30.5 ％),bcl-6 rearrangement in 11 cases (18.6 ％),5 cases with c-myc rearrangement (8.5 ％).In the aspects of remission rate,p53 deletion positive group contained less advantage than negative ones (33.3 ％ vs 75.6 ％,x2 =9.560,P =0.002).The prognosis of bcl-6 gene rearrangement positive group different from negative group,but the difference was not statistically significant (OS,P =0.107; PFS,P =0.094),p53 deletion positive patients was in significantly worse prognosis than the negative group (OS,P =0.031; PFS,P =0.028),c-myc rearrangement positive group difference in gene rearrangement negative group,but the difference was not statistically significant (OS,P =0.163; PFS,P =0.167).In the CHOP group,prognosis of p53 deletion,c-myc rearrangement positive group were significantly worse than the negative group,the difference was statistically significant (P ＜ 0.05).In R-CHOP group,the prognostic significance of bcl-6 gene rearrangement positive group were worse (OS,P =0.003; PFS,P =0.007).Conclusion DLBCL patients with bcl-6,p53,c-myc genes aberrations are related with poor prognosis,and they can be used as prognostic factors for predicting DLBCL and guiding therapy.

We investigated the effects of dendritic cell (DC) pulsed with acute leukemia cell frozen-thawed antigen on inducing the cytotoxic T lymphocyte (CTL) to get specific anti-tumor activity in vitro. DC was generated from healthy human bone marrow mononuclear cell (BMMC) in the presence of granulocyte/macrophage-colony stimulating factor(GM-CSF), interleukin-4 (IL-4) in vitro. DC pulsed with acute leukemia cell frozen-thawed antigen was co-cultured to induce T cell into specific CTL. Then we observed the effects of CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen killing acute leukemia cell specially and the influence of dendritic cell affecting the function and CD expression on CTL. The levels of CD1a, CD86, HLA-DR expression on DC pulsed with acute leukemia cell frozen-thawed antigen were obviously higher than those before culture (P<0.01). There were more CD3+CD8+ T cells in the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen, compared with those in the T cell uncultured group (P<0.01). The CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen significantly had higher activity in killing acute leukemia cell than in killing k562 cell (P<0.01), and the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen was also more effective for killing acute leukemia cell as compared with the CTL induced by DC simply, T cell co-cultured with IL-2 and T cell simply (P<0.01). The DC generated from human bone marrow mononuclear cell (BMMC) in the presence of granulocyte/macrophage-colony stimulating factor (GM-CSF), interleukin-4 (IL-4) was CD14- CD1a+CD83+DC, and it could also induce the cytotoxic T lymphocyte (CTL) to get specific anti-tumor activity in vitro. Otherwise,the increasing of CD3+CD8+ T cells in the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen implied the main role of the CD3+CD8+ T cells in the anti-tumor immunity.
Antigens, Neoplasm ; immunology ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interleukin-4 ; pharmacology ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Lymphocyte Activation ; T-Lymphocytes, Cytotoxic ; immunology