Apoptosis of leukemia Molt-4 cells induced by resveratrol and effect of WAVE1 gene expression / 白血病·淋巴瘤

Objective To investigate the possible mechanism of apoptosis effect induced by resveratrol in human acute T lymphoblast leukemia Molt-4 cells. Methods MTT method was used to analyze the inhibition rate. The cell cycle and apoptosis percentage of Molt-4 cells treated with resveratrol were detected by flow cytometry. The expression of WAVE1 mRNA was assessed by semi-quantitative RT-PCR. Results After treated with 12.5, 25.0, 50.0, 100.0, 200.0μmol/L resveratrol for 24, 48 and 72 hours, the inhibitory effects were at 29.32 %, 36.11 %, 53.92 %, 62.50 %, and 74.98 %, respectively. Resveratrol was able to inhibit the proliferation of Molt-4 cells in a time-dependent and dose-dependent manner (F =33.614, P <0.05).Compared with control group, after treated with 50.0, 100.0 μmol/L resveratrol for 48 h, the cell number in S phase of Molt-4 cells was 68.6 % and 78.1 %, respectively, and the cell cycle was arrested at S phase (F = 19.453, P < 0.01) after treated with 50.0, 100.0 μmol/L resveratrol for 48 h, the ratio of WAVE1/GAPDH was 0.356±0.03, 0.382±0.05. Compared with control group 0.586±0.06, the ratio of WAVE1/GAPDH was decreased (F =8.950, P <0.01). Conclusion Resveratrol could induce the cells to undergo apoptosis, which was probably related to downregulation the expression of WAVE1.

CD4+CD25high regulatory cells in peripheral blood of NSCLC patients / 华中科技大学学报（医学）（英德文版）

The proportion and changes of CD4+CD25high regulatory T cells (Trs) in peripheral blood of non-small cell lung cancer (NSCLC) patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets (CD3+, CD4+ and CD8+) and CD4+CD25high Tr cells. The results showed that the proportion of CD4+CD25high Tr cells in NSCLC group was significantly higher than in control group [(4.36 +/-2.07) % vs (2.04+/-1.03) %, P<0.01]. The proportion of CD4+CD25 high Tr cells in late stage was higher than that in early stage [stages I +II (2.26+/-0.6) %; stage III (3.28+/-1.38) %; stage IV (6.06 +/-4.08) %] (P<0.05). Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4+CD25high Tr cells in peripheral blood was worse (P=0.0026). In conclusion, the relative increase in CD4+CD25high Tr cells in peripheral blood may be related to immunosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4+CD25+high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4+CD25high Tr cell therapy to treat NSCLC patients may be an effective strategy.

CD4+CD25high Regulatory Cells in Peripheral Blood of NSCLC Patients / 华中科技大学学报（医学）（英德文版）

The proportion and changes of CD4+CD25high regulatory T cells (Trs) in peripheral blood of non-small cell lung cancer (NSCLC) patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets (CD3+, CD4+ and CD8+) and CD4+CD25high Tr cells. The results showed that the proportion of CD4+CD25high Tr cells in NSCLC group was significantly higher than in control group [(4.36±2.07) % vs (2.04±1.03) %, P＜0.01]. The proportion of CD4+CD25 high Tr cells in late stage was higher than that in early stage [stages Ⅰ + Ⅱ (2.26±0.6) %; stage Ⅲ (3.28±1.38) %; stage Ⅳ(6.06±4.08) %] (P＜0.05). Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4+CD25high Tr cells in peripheral blood was worse (P=0.0026). In conclusion, the relative increase in CD4+CD25high Tr cells in peripheral blood may be related to immunosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4+CD25+high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4+CD25high Tr cell therapy to treat NSCLC patients may be an effective strategy.

The effect of human cytomegalovirus on the formation of CFU-MK in vitro / 华中科技大学学报（医学）（英德文版）

To investigate the mechanism and the suppressive effect of human cytomegalovirus (HCMV) on colony forming unit-megakaryocyte (CFU-MK), semi-solid culture system was used to observe the effect of HCMV AD169 strain on CFU-MK's growth of 18 cord blood samples. HCMV DNA and immediate early (IE) protein mRNA in CFU-MK was detected by PCR and reverse transcription-polymerase chain reaction (RT-PCR). Our results showed that HCMV AD169 significantly suppressed the formation of CFU-MK in vitro. Compared with the mock group, the CFU-MK colonies decreased by 21.6%, 33.8% and 46.3%, respectively, in all the 3 infected groups (P<0.05), suggesting the suppression and the titer of the virus was dose-dependent. Both HCMV DNA and the expression of HCMV IE protein mRNA were positively detected in the colony cells of viral infected group. It is concluded that HCMV AD169 strain could inhibit the differentiation and proliferation of CFU-MK by directly infecting their progenitors. There was early transcription of HCMV IE protein in CFU-MK infected by virus.

The effect of costimulatory factors in the pathogenesis of chronic idiopathic thrombocytopenic purpura / 华中科技大学学报（医学）（英德文版）

To investigate the effect of costimulatory factors in the pathogenesis of chronic idiopathic thrombocytopenic purpura (CITP), we examined the expression of CD80 on platelets and megakaryocytes in patients with CITP and the controls by FACS. By using CD80 monoclonal antibody (McAb) to inhibit interaction among cells which is mediated by costimulatory factors, we observed the effect of CD80 McAb on the growth and maturation of megakaryocytic progenitors of patients with CITP in vitro. The results showed the expression of CD80 on platelets and megakaryocytes in CITP group was significantly higher than that in controls (P<0.01). There was a significantly positive correlation between the expression of CD80 on platelets and serum PAIgG in CITP (r=0.86, P<0.05). The mean of various clone numbers (CFU-MK, BFU-MK and mCFU-MK) in CITP were all lower than those in controls (P<0.05). In megakaryocytes co-cultured with CD80 McAb, there was an increasing tendency of the number of CFU-MK and big CFU-MK (the number of megakaryocyte with GP IIIa positive was more than 20) and mediate CFU-MK (the number of megakaryocyte with GP IIIa positive was 11-20). When the concentration of CD80 McAb was 10 microg/L, there was a significant difference in the number of megakaryocytic colony formation (CFU-MK, BFU-MK and mCFU-MK) between the group with CD80 McAb and that without it (P<0.05). These showed the abnormality of costimulatory factors had important effect in the pathogenesis of CITP.

CD4~+CD25~+/CD4~+CD25~(high) regulatory cells in peripheral blood of cancer patients / 中国免疫学杂志

Objective: To investigate the frequencies of CD4+ regulatory T(Treg) cells(CD4+CD25+/CD4+CD25high) in peripheral blood of cancer patients,providing the opportunity to determine whether cancer patients exhibit an expanded CD4+CD25high pool. Methods: The frequency of CD4+CD25+/CD4+CD25high Treg in the peripheral blood of 62 cancer patients and 15 controls was determined by flow cytometry. Results: Compared with those of healthy control, the frequency of CD4+CD25+/CD4+CD25high in the peripheral blood of cancer patients showed a significant increase[(19.61?8.17)%/(4.20?1.90)%,P