Expression of caspase-4 in Hodgkin's lymphoma and anaplastic large cell lymphoma

AIM: This study is based on the result of the study in HL and ALCL employing gene chip technique, in which writer found that there was distinctly different expression of caspase-4 between HL and ALCL cell lines at the level of mRNA. From the point of view, we try to identify at the level of protein whether there is different expression of this gene in HL and ALCL tissues as well. METHODS: HE staining, the monoclonal antibodies CD30 (BerH2), CD15 (C3D-1), CD20 (L26) and CD45RO (UCHL1) were used for selecting the cases of HL and ALCL. Specific high affinitive anti-caspase-4 polyclonal antibody was used by immunohistochemical staining to analyze the expression of caspase-4 in 18 cases of HL and 15 cases of ALCL. RESULTS: The expression of caspase-4 demonstrated a strong positive staining in all ALCL cases (15/15,100%), whereas negative in 16 HL cases (88.8%), while other two cases were weakly stained (11.2%), showing a distinct difference (P<0.01) between two groups. CONCLUSIONS: (1) The diverse expression of caspase-4 gene in HL and ALCL groups implies a different mechanism of oncogenesis and the different defects of signal transduction pathway of apoptosis in these two entities of lymphomas. (2) Clarification of this gene might be useful for the differential diagnosis of HL and ALCL. Furthermore, it probably provides theoretically gene therapy strategies for lymphoma in the future.     

Expression of mic2/CD99 protein and their correlation with Eber-1/LMP-1 in Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma

AIM: To study the expression of mic2/CD99 protein and their correlation with Eber-1/LMP-1 in Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma. METHODS: Immunohistochemical staining, in situ hybrization and tissue microarry technique were used to detect the expressions of mic2/CD99 and Eber-1/LMP-1 of H/RS cells in 43 cases of cHL and 16 cases of NHL. RESULTS: The positive rate of CD99 protein expression in 43 cases of cHL was 2.3％ (1/43), mic2 was 55.8％ (24/43), LMP1 was 58.1％ (25/43) and Eber-1 was 53.5％ (23/43). The expressions of CD99 and mic2 in the NHL group were higher than those in cHL group (P<0.05). The expressions of LMP1 and Eber-1 in NHL group were lower than those in cHL group (P<0.05). The expression of mic2 was higher than CD99 in cHL group (P<0.05). There was no difference between the expression of Eber-1 and LMP1 in cHL group statistically (P>0.05). There was a negative correlation between the expression of CD99 protein and LMP1 in H/RS cells (P<0.05). No correlation was found between the expression of any other markers in H/RS cells and patients' sex (P>0.05). There was a significant correlation between the high expression of LMP1 and a low expression of CD99 in the young patients (P<0.05), but no correlation was found between the expression of mic2 and Eber-1 in young patients (P>0.05). CONCLUSION: There is a negative correlation between the expression of LMP1 and CD99 in Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma.     

Angiogenic effect of HIF-1α on extranodal nasal-type NK/T-cell lymphoma

AIM: To study the angiogenic effect of hypoxia inducible factor 1α(HIF-1α) and its significance on human extranodal nasal-type NK/T-cell lymphoma. METHODS: The protein levels of HIF-1α, vascular endothelial growth factor(VEGF) and VEGF receptor 2(VEGFR2) in human extranodal nasal-type NK/T-cell lymphoma were detected by immunohistochemistry. Microvessel density (MVD) of the tumor tissues was determined by labeling of microvessel endothelium with CD34 antibody. The correlation between the expression of HIF-1α, VEGF and VEGFR2 and MVD was analyzed with SPSS 13.0 statistical software. RESULTS: The positive expression of HIF-1α was observed in 39 cases (39/50, 78%) and the positive expression of VEGFR2 was 27 cases (27/50, 54%) of human extranodal nasal-type NK/T-cell lymphoma. A statistical difference of HIF-1α and VEGFR2 expression between tumor tissues and normal lymphocytes in lymph node was observed (P<0.05). In the tumor tissues, the co-expression of VEGF or VEGFR2 with HIF-1α was 72% and 64%, respectively, significantly higher than that without HIF-1α co-expression (P<0.05). The expression of HIF-1α, VEGFR and VEGFR2 was positively correlated with MVD of the tumor tissues (P<0.01). HIF-1α was expressed in all 15 cases of extranodal nasal-type NK/T-cell lymphoma with angiocentric infiltration.CONCLUSION: HIF-1α may promote angiogenesis of extranodal nasal-type NK/T-cell lymphoma through VEGF/VEGFR2 signaling pathway.     

Detection of CD23 mRNA expression in B-cell lymphoma by in situ hybridization with digoxigenin-labelled DNA probe

AIM: To detect CD23 mRNA expression in B-cell lymphoma. METHODS: 20 pathological diagnostic conformed B-cell lymphoma samples were selected. Surface CD23 protein expression was examined by SP-immunohistochemistry and CD23 mRNA was detected by in situ hybridization with digotoxin labeled CD23 DNA probe.RESULTS: The positive rates of both surface CD23 protein and CD23 mRNA expression in B-cell lymphoma samples were more than 90%. CONCLUSIONS: The high expression of CD23 was showed in B-cell lymphoma both in mRNA and in protein levels. The results of this study was useful for understanding the molecular mechanism of B-cell lymphoma and for clinical diagnosis and treatment of the disease.     

Study on genetic instability of nm23-H1 gene and expression of hMLH1,hMSH2 in sporadic gallbladder carcinoma

AIM: To examine the MSI and LOH of locus D17S396 and their influence on the expression of nm23-H1 in gallbladder tumors,and to examine the protein expression of hMLH1/hMSH2,which may provide experimental evidence for the tumor occurrence and metastasis.METHODS: Techniques such as DNA extraction,CR-SSCP,ordinary silver stain were used to study MSI and LOH of locus D17S396.Envision IHC was used to assess the expression of nm23-H1 and hMLH1/hMSH2.RESULTS: ① The frequency of heredity instability of gallbladder carcinoma was 42.55%.The frequency of LOH in liver and lymph node metastasis cases and in stage Nevin IV and V was significantly higher than that without metastasis and stage I,II and III.However,the frequency of MSI showed contrary correlation with some clinicopathologic characteristics.② The expression of nm23-H1 was 46.81%.The case with lymph node metastasis and Nevin stage IV and V showed significantly lower expression than that without lymph node metastasis and stage I,II and III.③ The expressions of hMLH1 and hMSH2 were 51.06% and 42.55% respectively.hMLH1 in lymph node and liver metastasis cases and in stage Nevin IV and V were significantly lower than that without metastasis and in stage I,II and III.④ Positive frequency of hMLH1 in MSI positive group was higher than that in MSI negative group.The positive frequency of nm23-H1 and hMSH2 protein in LOH positive group was lower than that in negative group.CONCLUSION: The heredity instability of nm23-H1 gene may be implicated pathogenesis and progression of gallbladder carcinoma.Both MSI and LOH of nm23-H1 control the development of gallbladder carcinoma independently in different paths.Abnormal expression of hMLH1/hMSH2 may be a molecule marker in early stage of gallbladder carcinoma.     

Over-expression of hsa-miR-150 induces re-differentiation of diffuse large B-cell lymphoma cell line OCI-Ly10

AIM:To investigate the mechanism that over-expression of hsa-miR-150 induces the re-differentiation of diffuse large B-cell lymphoma cell line OCI-Ly10. METHODS:The expression level of hsa-miR-150 in CD19+B and OCI-Ly10 cell lines was detected by real-time PCR. The expression level of c-Myb was detected by Western blotting and immunofluorescence cytochemistry methods. Lentiviral supernatant containing recombinant plasmids was transfected into OCI-Ly10 cells by LipofectamineTM 2000 and named Ly10-control and Ly10-miR-150. The biological functions of the 2 cell sublines were identified by MTT assay. The cell cycle and apoptotic rates were detected by flow cytometry. The expression levels of B-lymphocyte differentiation-related genes and c-myb in Ly10-control and Ly10-miR-150 cells were detected by real-time PCR and Western blotting. When c-myb was interfered in by interference fragment in OCI-Ly10 cells, the interference efficiency and the expression levels of BCL6 and PRDM1 were detected by real-time PCR and Western blotting. RESULTS:The expression level of hsa-miR-150 in CD19+ B cells was significantly higher than that in OCI-Ly10 cells. The expression level of c-Myb in OCI-Ly10 cells was higher than that in CD19+ B cells. The expression levels of B-lymphocyte differentiation-related genes were changed significantly in OCI-Ly10 cells after transfected with hsa-miR-150. The expression levels of PAX5, BCL6 and c-Myb in Ly10-miR-150 cells were lower than those in Ly10-control cells, but the expression levels of IRF4, PRDM1 and XBP1 were higher than those in Ly10-control cells. The expression level of BCL6 was lower and PRDM1 was higher after interference. CONCLUSION:Hsa-miR-150 plays a significant role in inhibiting proliferation and inducing apoptosis of OCI-Ly10 cells. The mechanism that over-expression of hsa-miR-150 induces OCI-Ly10 cell differentiation toward terminal B cells may be related to the down-regulation of c-myb.     

Effects of ischemic preconditioning on cardiac myocyte apoptosis and expression of bcl-2 during myocardial ischemia/reperfusion in rats

AIM:To explore the effect of ischemic preconditioning on cardiac myocyte apoptosis and the expression of bcl-2 during myocardial ischemia/reperfusion in rats. METHODS:We use TUNEL,immunohistochemical and in situ hybridization(ISH) methods to detect the cardiac myocyte apoptosis and the expression of bcl-2 during myocardial ischemia/reperfusion in rats. RESULTS:①The numbers of positive cardiac myocyte nuclear and the percentage of positive cardiac myocyte nuclear in IP+I/R_3h group decreased significantly(P<0.05,P<0.01)compared with I/R_3h group,respectively.②The numbers of bcl-2 protein positive cardiomyocyte and the percentage of bcl-2 protein positive cardiomyocyte in IP+I/R_3h group were higher(P<0.01)than that of I/R_3h group,respectively.The numbers of positive bcl-2 mRNA cardiomyocyte and the percentage of positive bcl-2 mRNA cardiomyocyte in IP+I/R_1h group were higher(P<0.01)than that of I/R_1h group,respectively.CONCLUSION:① The first window of IP's protection could reduce cardiomyocyte apoptosis significantly.② Up-regulating the protein expression of bcl-2 in cardiomyocytes during I/R may be one of the mechanisms of first window of IP's protection.     

Hypoxia upregulates CD73 expression in mouse microvascular endothelial cells

AIM: To study the effect of hypoxia on CD73 expression in mouse microvascular endothelial cell line bEnd.3. METHODS: ① bEnd.3 cells were exposed to different periods of hypoxia. ② Concentration of LDH released by bEnd.3 cells into the culture medium was detected. ③ Surface CD73 activity in bEnd.3 cells was measured by HPLC according to the conversion of E-AMP to E-ADO. ④ CD73 mRNA expression were analyzed by semiquantitative RT-PCR. ⑤ Cell surface proteins were biotinylated and CD73 was detected by avidin blots of immunoprecipitation with mAb TY23. RESULTS: ① bEnd.3 cells exposed to hypoxia for 24 h demonstrated a significant increase in LDH release (P<0.01). ②Hypoxia induced CD73 activity in bEnd.3 cells in a time-dependent manner. ③ CD73 mRNA expression increased markedly in hypoxia for 4 h and 8 h (P<0.05). ④ bEnd.3 cells exposured to hypoxia induced a time-dependent increase in expression of CD73 (P<0.05). CONCLUSION: Hypoxia induces CD73 mRNA, protein expression and increases CD73 activity in mouse microvascular endothelial cells.     

Relationship between genetic instability of nm23H1 gene and clinical pathological characteristic in hepatocellular carcinoma

AIM: The aim of this study was to examine the microstatellite instability (MSI) and loss of heterozygosity(LOH) of locus D17S396 on chromosome 17 and their influence on the expression of nm23H1 in hepatocellular carcinoma (HCC),which may provide experimental evidence for the mechanism of nm23H1 gene and tumor metastasis.METHODS: Techniques such as DNA extraction from formalin-fixed paraffin-embedded tissues,PCR-SSCP,ordinary silver stain were used to study MSI and LOH of locus D17S396.Envision immunohistochemistry and Leica-Qwin computer imaging techniques were used to assess the expression of nm23H1.RESULTS: ① The frequency of heredity instability of HCCs was 35.42%.The frequency of LOH in the cases with lymph node or distant organs metastasis or not and with intrahepatic metastasis or embolus of portal vein or not was significantly different (P<0.01),it was higher in stage TNM Ⅲ than that in stageⅠ and Ⅱ.Moreover,it was higher in high tendency to invasion or metastasis cases than that in the low tendency cases (P<0.01).② The expression of nm23H1 was 56.25%.It was significantly different in Edmondson grade,TNM stage and in lymph node or distant organ metastasis cases (P<0.01).The cases with high tendency of invasion or metastasis exhibited lower nm23H1 expression compared with low tendency cases (P<0.01).③ The positive rate of nm23H1 protein in LOH positive group was lower than that in LOH negative group (P<0.05).CONCLUSION: The results indicate that both MSI and LOH of nm23H1 gene control the development of HCC independently in different pathways.LOH inhibits the expression of nm23H1,which endows it with high aggressive and poor prognosis.Increase in the amount of nm23H1 protein expression effectively restrains the tendency to invasion or metastasis of HCCs and improves prognosis of patients.     

Effect of γ radioactive stent on proliferation and apoptosis of smooth muscle cells

AIM: To observe effect of γ radioactive ［103Pd］ stent on the proliferation and apoptosis of smooth muscle cells, the mechanism of radioactive stent preventing in-stent restenosis was explored. METHODS: Fifty male New Zealand rabbits were randomized into stent group and ［103Pd］ stent group. Control group was set up. The materials were harvested on 3, 7, 14, 28, 56 days after operation and the following investigation were carried out, including pathomorphology, immunohistochemistry, apoptosis (TUNEL) and in situ hybridization studies.RESULTS: ① The severity of the stenosis in ［103Pd］ stent group was less severe than that in stent group. It was most obvious on 56 th day (P<0.01). ② The expression of proliferating cell nuclear antigen(PCNA) of ［103Pd］ stent group was lower than that in stent group on 3 to 28 days. It was most obvious on 7th day, 16.35%±0.79% vs 24.36±0.55% (P<0.01). ③ TUNEL method showed that the ［103Pd］ stent group had much more apoptosis of VSMC than that in stent group. The highest rate of apoptosis appeared on day 7, 14.72%±0.53% vs 12.42%±1.13% (P<0.01). ④ By calculating the ratio of PCNA/apoptosis (P〖KG*6〗∶〖KG-*2〗A), a much lower ratio was seen in ［103Pd］-stent groups than that in stent group at 3 to 28 days. There was significant statistic difference (P<0.05). ⑤ For bcl-2/bax ratio, the result in ［103Pd］-stent group was lower than that in stent group at 3 to 28 days. There was significant statistic difference (P<0.05).CONCLUSION: γ radioactive stent inhibits the proliferation and accelerates apoptosis of injured media vascular smooth muscle cells. It decreases the ratio of proliferation to apoptosis and relieves the severity of restenosis.     

Apoptosis induced by 5-fluorocytosine in human pancreatic cancer cells genetically modified to express cytosine deaminase

AIM:To elucidate the pattern of 5-fluorocytosine(5-FC) induced apoptosis and its role in gene therapy for human pancreatic cancer. METHODS: The human pancreatic cancer SW1990 cells(CEA-producing) were infected with recombinant adenoviruses(Adex1CEA-prCD or Adex1CEA-prZ).Cytosine deaminase(CD) expression was examind by western blot. Apoptosis induced by 5-FC in human pancreatic cancer SW1990 cells genetically modified to express cytosine deaminase was investigated by applying electron microscopy, DNA electrophoresis and flow cytometry analysis techniques. RESULTS: The SW1990 cells infected with Adex1CEA-prCD were treated with 5-FC at 100 μmol·L^-1 for 48 h, cell apoptosis occurred. Typical apoptosis morphological feature appeared and DNA ladder could be demonstrated on DNA electrophoresis. Apoptosis peak was also showed by flow cytometry. Apoptotic cells accounted for 34.6% of the cell population. Cells in G₁, S and G₂/M phase of cell cycle were 64%, 11% and 7%, respectively. CONCLUSION: The apoptosis induced by 5-FC may be a primary mechanism in CD gene therapy for pancreatic cancer.     

IL-12 induces T lymphocytes underging apoptosis and affects the expression and signal conduction of Bcl-2

AIM: IL-12 acts upon T lymphocytes and activates its receptor complexes of β1/β2 ,and so IL-12 can regulate TH1/TH2 balance. Our study is aimed at IL-12-inducing apoptosis of T cells and expression and signal conduction of Bcl-2 during T cell apoptosis. METHODS: The apoptosis of T cells was detected by Annexin V staining cytometry and the expression of Bcl-2 under different inhibitors were detected by the method of semi-quantitative PCR. RESULTS: IL-12 can induce the human leukemic T cell line(TIB-152) and the human lymphoma T cell line(HTB-176) and the normal human T cells to undergo apoptosis. The Bcl-2 expression at 6 hours of treatment with IL-12 increased aparently, and reached the max at 24 hours. But IL-12 did influence Bcl-2 expression. IL-12 can induce T cells to undergo apoptosis which is characterized by early membrane changes. CONCLUSION: The inducing effect is correlated with the concentration of IL-12 and the maturation of T cells. Bcl-2 takes part in the progression of T cells' apoptosis as a apoptosis mediator.     

Change of calcium sensing receptor expression and apoptotic pathways in myocardial infarction rat induced by isoprel

AIM:To observe the change of calcium sensing receptor (CaSR) expression and apoptotic pathways in myocardial infarction rat induced by isoprel.METHODS: The myocardial infarction rat models were prepared by subcutaneous injection of isoprel (ISO 200 mg·kg-1·d-1 for 2 d). Wistar rats were divided into three groups randomly: ① Control group; ② ISO/1d group; ③ ISO /2d group. The expressions of CaSR, 〖STBX〗bcl-2, bax〖STBZ〗 and caspase-3 mRNA and protein were analyzed by RT-PCR and Western blotting, respectively. Apoptotic cells were measured by TUNEL staining assay. The morphological and ultrastructural changes were observed under optical microscope and electronic microscope. The activity of LDH, CK, SOD and the content of MDA were assayed with ultraviolet spectrophotometer. The level of troponin(cTnT) was observed by chemical immunofluoresence.RESULTS: Compared with control group, the activity of LDH and CK, the content of MDA and cTnT, the apoptosis index and the expression of CaSR, Bax and caspase-3 were reached the highest level, but the SOD activity and Bcl-2 expression were decreased. The myocardial ultrastructural injury was aggravated in the ISO/1d group. The change of above parameters in ISO/2d group was between control and ISO/1d group.CONCLUSION:The increased expression of CaSR is involved in rat myocardial infarction induced by isoprel, which is related with oxidative stress and apoptosis.     

Detection of monoclonal immunoglobulin heavy chain gene rearrangements and its application in diagnosis of B-cell non-Hodgkin lymphoma

AIM: To establish a reliable and feasible protocol for detection of monoclonal immunoglobulin heavy chain (IgH) gene rearrangements for routine diagnosis of B-cell non-Hodgkin lymphoma (B-NHL). METHODS: Using the primer combinations of FR2, FR3, LJH and VLJH, mode tube A+ tube B, and semi-nested PCR, the monoclonal IgH gene rearrangements in 121 cases of B-NHL, 58 cases of T-cell non-Hodgkin lymphoma (T-NHL) and 19 cases of reactive lymphoid hyperplasia were detected. The differences of clonality detection rate between B-NHL group and T-NHL group, B-NHL group and reactive lymphoid hyperplasia group, and between the use of FR2, FR3 and FR2+FR3 primers were analyzed. RESULTS: The clonality detection rates of B-NHL, T-NHL and reactive lymphoid hyperplasia were 81% (96/118), 4% (2/54) and 0% (0/19). There were remarkable differences between B-NHL group and T-NHL group, B-NHL group and reactive lymphoid hyperplasia group in monoclonal IgH gene rearrangements (P<0.05). In B-NHL group, monoclonality was found in 58% of the cases using primer FR2, 55% using FR3, and 81% using the combination of both primers, with significant differences (P<0.05). CONCLUSION: Using the primer combinations of FR2, FR3, LJH and VLJH, detection of paraffin-embedded tissues, the method of tube A+tube B mode and semi-nested PCR for determining monoclonal IgH gene rearrangements is feasible and reliable, and the clonality detection rate is high enough for clinical diagnosis of B-NHL.     

Expression of MMP-2 mRNA and MMP-9 mRNA in pulmonary arterioles ofrats with pulmonary hypertension induced by chronic hypoxia hypercapnia

AIM:To investigate the expression of matrix metalloproteinases(MMPs) in pulmonary arterioles of rats with chronic hypoxia and hypercapnia-induced pulmonary hypertension.METHODS:MMP-2, MMP-9 and MMP-2 mRNA, MMP-9 mRNA were observed in pulmonary arterioles by the techniques of immunohistochemistry and in situ hybridization.RESULTS:①The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) of hypoxia-hypercapnia groups were higher than those of normal control group (P＜0.01). ②Light microscopy showed that vessel wall and media of pulmonary arterioles were thicker in rats of hypoxia-hypercapnia groups than normal control group. There were vessel smooth muscle cell hypertrophy, vessel cavity straitness in hypoxia-hypercapnia group, but no same performance was found in normal control group. ③The expression of MMP-2, MMP-9 and MMP-2 mRNA, MMP-9 mRNA in pulmonary arterioles were significantly higher in rats of hypoxia-hypercapnia groups than control group (P＜0.01).CONCLUSION:Expression of matrix metalloproteinases in pulmonary arterioles is enhanced by hypoxia hypercapnia. This may be involved in pulmonary vascular remodeling in rats with pulmonary hypertension.     

Expression of cytokines and their receptors in leukemia cell lines and normal blood cells

AIM: To study the expression of cytokines and their receptors in leukemia cell lines and normal blood cells. METHODS: RT-PCR was used to detect expression of mRNA for cytokines in leukemia cell lines(HL-60,U937,K562,HEL,DAMI,MEG-01,HUT78 and CA) and normal blood cells, including CD34⁺ cells, megakaryocytes,platelets, peripheral mononucleates cells and granulocytes. RESULTS: ①CD34⁺ cells simultaneously expressed mRNA for IL-1(α,β),IL-3, IL-6 , G-CSF, GM-CSF and their receptors and SCFR,MPL as well. The granulocytes only expressed IL-6,IL-6R,G-CSFR,GM-CSF. Megakaryocytes and platelets only expressed IL-3R,IL-6,IL-6R,MPL.Interestingly, TGFβ₁ ,TNFα and their receptors sustained to express in normal cells.②Most leukemia cell lines were found to simultaneously express at least two or more stimulating cytokines and receptors ,while TGFβ 1 , TNFα and their receptors were expressed in all the leukemia cell lines we observed. CONCLUSIONS: ①Multi-autocrine loops exist in leukemia cells;②Imbalance of autocrine loops of positive and negative cytokines may be related to leukemia.     

Plumbagin enhances TRAIL-induced apoptosis in Kasumi-1 cells

AIM: To investigate the effect of plumbagin and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the apoptosis of leukemic Kasumi-1 cells. METHODS: Kasumi-1 cells were treated with plumbagin alone, recombinant soluble TRAIL(rsTRAIL) alone or the combination of plumbagin with rsTRAIL to induce apoptosis. The cell proliferation was analyzed by CCK-8 assay. Apoptosis was determined by flow cytometry with AnnexinⅤ/PI double staining and TUNEL staining. The expression of DR4 and DR5 at mRNA level was measured by real-time PCR. The expression of signal transduction proteins, such as DR5, caspase-3, caspase-8, caspasep-9, Bid, Bax and c-FLIP was detected by Western blotting. RESULTS: Both rsTRAIL and plumbagin induced the apoptosis in Kasumi-1 cells, and combination of plumbagin with rsTRAIL enhanced the apoptosis. The ratios of Annexin V-positive Kasumi-1 cells were (27.7±2.9)%, (25.6±3.1)% and (52.1±3.3)% in 100 μg/L rsTRAIL group, 2 μmol/L plumbagin group and the combination group, respectively, and the positive rate in combination group was significantly higher than those in other 2 groups. TUNEL assay demonstrated that the number of apoptotic cells in combination group was higher than that in the cells treated with rsTRAIL or plumbagin alone. Plumbagin up-regulated the expression of DR5 at mRNA level in Kasumi-1 cells, and up-regulation of DR5, activation of caspase-8 and down-regulation of c-FLIP at protein level were detected in the cells treated with plumbagin alone and the combination of plumbagin with rsTRAIL. CONCLUSION: Plumbagin enhances TRAIL-induced apoptosis in Kasumi-1 cells by up-regulating DR5, activating caspase-8 and down-regulating c-FLIP.     

Expression of CD44s in lung cancer and its significance

AIM: To investigate expression of CD44s in lung cancer and it's clinical significance. METHODS: A total of 117 primary lung cancer from patients were examined for CD44s expression by immunohistochemical staining. RESULTS: CD44s mostly expressed in non-small cell lung cancer (NSCLC) but not in small ecll lung cancer (SCLC), and squamous cell carcinoma(SCC) showed much stronger expression of CD44s than adenocarcinoma(ADC)(P＜0.05). In comparison of the lung cancer with/ without lymph node metastasis, the latter showed stronger expression of CD44s(P＜0.01). According to TNM, there was a distinct statistic difference between early stage and advanced stage(P＜0.05). CONCLUSION: CD44s might be a better indicant in histological classification of lung cancer, lymph node metastasis, clinical stage and prognosis.     

CpG-oligodeoxynucleotides induce differentiation and apoptosis in HL60 cells

AIM: To investigate the differentiative and apoptotic effect of CpG-oligodeoxynucleotides on HL60 cells and its mechanism. METHODS: After HL60 cells were exposed to synthetic CpG-oligodeoxynucleotides,non-CpG-oligodeoxynucleotides or ZpG-oligodeoxynucleotides for 72 hours, respectively, the inhibition of HL60 cells were detected using MTT method, NBT test was used and CD14 expression were determined.,Apotosis of HL60cells were mensurated with flow cytometry and transmission electron microscope, and caspase 3，Bcl-2 and Bax expres-sion of HL60 cells treated with oligodeoxynucletides were detected using immunohistochemistry RESULTS: Treatment with CpG-oligodeoxynucleotides induced the differentiation and apoptosis in HL60 cells, but non-CpG-oligodeoxynucleotide and ZpG-oligodeoxynucleotide had no effect on HL60 cells. CONCLUSION: CpG-oligodeoxynucleotides can induce the differentiation and apoptosis in HL60 cells. It may provide a new approach for the immunological treatment of leukemia.     

Determination of T cell cycle and the expression of bcl- 2 in asthmatic mice and its significance

AIM: To investigate the changes of T cell cycle, the expression of bcl- 2 in allergic asthmatic mice and the effects of dexamethasone on them. METHODS: An animal model with asthma was established by means of ovalbumin sensitizing-challenging. CD3 expression in spleen and lymphocytes in bronchoalveolar lavage fluid (BALF), T cell cycle and Bcl-2 expression in spleen were detected by flow cytometry. RESULTS: In BALF lymphocytes and spleen lymphocytes, CD3 expression rate in the asthmatic group was significantly higher than that of control group. In BALF lymphocytes, CD3 expression rate in the asthma plus dexamethasone group was significantly lower than that of the asthmatic group. However, in spleen lymphocytes, CD3 expression rate in the asthma plus dexamethasone group was significantly higher than that of the asthmatic group. In spleen lymphocytes, the cell count in S phase, G₂+M phase and apoptosis rate of T cell from the asthmatic group were significantly higher than that from the control group. Cell count in S phase, G₂+M phase and apoptosis rate of T cell from the asthmaplus dexamethasone group was significantly lower than that from the asthmatic group. The Bcl-2 expression rate of T cell from the asthmatic group was significantly higher than that from the control group. CONCLUSIONS: In the allergic asthmatic mice model, T cell count, proliferation and activation of T cells, apoptosis rate of T cells in spleen lymphocytes increase, meanwhile bcl- 2 expression also increases significantly. There was no significant effect of dexamethasone on the bcl- 2 expression. The therapeutic effects of dexamethasone on asthma may be not due to the inhibition of the bcl- 2 expression in T cells.