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Objective: To investigate the effects of curcumin combined with thalidomide on the proliferation and apoptosis of acute myeloid leukemia KG-1 cells, and its correlation with B-cell lymphoma-xL (Bcl-xL), signal transducer and activator of transcription 3 (STAT3)., Methods: MTT assay was used to detect the proliferation of KG-1 cells and screen the optimal combined concentration of curcumin and thalidomide. The effects of curcumin, thalidomide and their combination on the proliferation and apoptosis of KG-1 cells were analyzed by MTT method and flow cytometry, respectively. The mRNA expression levels of STAT3 and Bcl-xL in single-drug group, two-drug combination group and control group (untreated cells) were detected by real-time quantitative PCR., Results: Both curcumin and thalidomide inhibited the proliferation of KG-1 cells in a concentration-dependent manner in the range of 20-100 μmol/L ( r =0.657, r =0.681). The IC 50 value of curcumin and thalidomide at 48 h was (42.07±0.50) μmol/L and (57.01±2.39) μmol/L, respectively. The cell proliferation inhibition rate of curcumin (40 μmol/L) + thalidomide (60 μmol/L) was (86.67±1.53)%, which was significantly higher than (51.67±1.15)% of curcumin (40 μmol/L) and (55.33±1.53)% of thalidomide (60 μmol/L) (both P < 0.05). Treated with curcumin and thalidomide alone or in combination, the apoptosis rate of KT-1 cells was (18.67±2.08)%, (21.33±2.52)%, and (46.67±1.53)%, respectively, which was significantly higher than (0.72±0.03)% of control group (all P < 0.05). The cell apoptosis rate of two-drug combination group was significantly higher than that of single-drug group (both P < 0.05). Compared with the control group, the mRNA expressions of STAT3 and Bcl-xL in single-drug group, two-drug combination group were significantly decreased (both P < 0.05). Compared with single-drug group, the mRNA expressions of STAT3 and Bcl-xL in two-drug combination group were also significantly decreased (both P < 0.05)., Conclusion: Curcumin combined with thalidomide can synergistically down-regulate the expression of STAT3 and Bcl-xL, inhibit the proliferation of KG-1 cells, and induce apoptosis.

Objective: To investigate the effect of overexpression of leukemia inhibitory factor (LIF) on cisplatin and paclitaxel resistance of endometrial cancer cells in vitro ., Methods: Endometrial cancer cell lines HEC-1B and RL95-2 were infected with a recombinant lentivirus to overexpress LIF, and the changes in LIF expression was verified using RT-qPCR and ELISA. The viability of the LIF-overexpressing cells was assessed using CCK-8 assay, and the cell apoptosis and changes in mitochondrial membrane potential in response to cisplatin or paclitaxel treatment were analyzed with annexin V-FITC/PI staining and JC-1 assay, respectively. The effect of LIF overexpression on the expressions of Bcl-2 family proteins and STAT3 pathway was evaluated using Western blotting; dual-luciferase reporter gene assay was employed to detect the transcriptional activity of STAT3. The effect of STAT3 silencing on apoptosis of the LIF-overexpressing cells induced by cisplatin or paclitaxel was investigated., Results: The cell lines infected with the recombinant lentivirus showed significantly increased mRNA and protein levels of LIF ( P < 0.05) without obvious changes in the cell viability ( P >0.05). LIF overexpression significantly attenuated cisplatin-or paclitaxel-induced apoptosis of the endometrial cancer cells ( P < 0.05) and markedly increased mitochondrial membrane potential of the cells ( P < 0.05). The expressions of Bcl-2, Bcl-xL and p-STAT3 proteins increased obviously while the expressions of Bax, Bad and STAT3 either decreased or showed no obvious changes in the LIF-overexpressing cells. Overexpressing LIF significantly enhanced the transcriptional activity of STAT3 ( P < 0.05), and silencing STAT3 obviously enhanced apoptosis of the endometrial cancer cells overexpressing LIF ( P < 0.05)., Conclusions: s Overexpression of LIF can enhance cisplatin and paclitaxel resistance to endometrial cancer cells in vitro .

Objective: To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion. Methods: Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF. Results: CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group ( P <0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group ( P <0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group ( P <0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other ( P <0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group ( P <0.05). Conclusions: Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.

Objective To investigate the effect of interferon-β (IFN-β) combined with all-trans retinoic acid (ATRA) on the proliferation and apoptosis of HepG2 human hepatocarcinoma cells and the role of Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signal pathway in the process. Methods HepG2 cells were randomly divided intro three groups and treated with 1000 U/mL IFN-β, 10 μmol/L ATRA and 1000 U/mL IFN-β combined with 10 μmol/L ATRA, respectively for 24 hours. Cell viability was measured by MTT assay and apoptosis rate was detected by flow cytometry. Western blotting was applied to detect the protein levels of p-JAK2, p-STAT3, gene associated with retinoid-interferon-induced mortality-19 (GRIM-19), Bcl-2, Bcl-xl and Bax. Results IFN-β or ATRA inhibited the proliferation and induced the apoptosis of HepG2 cells. The effect was enhanced when IFN-β was combined with ATRA. The expressions of p-JAK2 and p-STAT3 were down-regulated while the expressions of GRIM-19 and Bax were up-regulated after treated with IFN-β or ATRA on HepG2 cells, especially the combination of IFN-β and ATRA. Conclusion Combination of IFN-β and ATRA could suppress the proliferation and induced the apoptosis of HepG2 hepatocarcinoma cells by inhibiting JAK2/STAT3 signal pathway.

Objective: To investigate the expressions of Bcl-xl and caspase-3 in the tissues of benign prostatic hyperplasia(BPH) and prostate cancer(PCa) and their correlation., Methods: Using immunohistochemical methods, we determined the expressions of Bcl-xl and caspase-3 in 20 cases of BPH and 35 cases of PCa and analyzed their correlation., Results: The positive rates of Bcl-xl in the BPH and PCa tissues were 45. 00% and 88. 57%,and those of caspase-3 were 60. 00% and 17. 14%,respectively. The expressions of Bcl-xl and caspase-3 in the PCa tissue were not correlated with the preoperative PSA level, the risk of distant metastasis, Gleason scores, and clinical stages( P > 0. 05),and there was a negative correlation between the expressions of the two proteins in PCa( rs=- 0. 748,P < 0. 05)., Conclusion: The expression of Bcl-xl was upregulated while that of caspase-3 downregulated significantly in the PCa tissue, which indicates a possible interaction between the two proteins in cell apoptosis.

Objective: To study the effect of reactive oxygen species (ROS) on the regulation of platelet apoptosis., Methods: Washed healthy human platelets were pre-incubated with N-caetyl-Lcysteine (NAC), and then stimulated with dibucaine or thrombin. The production of ROS and depolarization of mitochondrial membrane potential (∆ ψm) were detected by flow cytometry. The activation of caspase-3 and expression of Bcl-xL were analyzed by Western blot., Results: (1)The average ROS fluorescence value of NAC+dibucaine group was lower than that of dibucaine group(0.66 ± 0.11 vs 1.06 ± 0.08, P<0.01), while that of NAC+thrombin group was also lower than that of thrombin group(0.45 ± 0.05 vs 0.71 ± 0.11, P=0.001). (2)The percentage of platelets with normal ∆ψm in NAC+Dibucaine group was higher than that of dibucaine group[(86.30 ± 9.37)% vs (13.52 ± 3.01)%, P=0.000], while that of NAC+thrombin group was also higher than that of thrombin group[(93.00 ± 3.03)% vs (76.58 ± 5.28)%, P=0.000]. (3)Fragmentation generated by caspase-3 activation in dibucaine group was much more than that in DMSO control group, while the fragmentation in NAC+dibucaine group was significantly decreased. (4)The expression of anti-apoptosis protein Bcl-xL of NAC+dibucaine group was significantly higher than that of the dibucaine group, while that of NAC+thrombin group was also higher than that of thrombin group., Conclusion: Through the regulation of ROS, NAC could inhibit the platelet apoptosis induced by dibucaine or thrombin.

Objective: To evaluate the anti neoplastic effects of p21(WAF1/CIP1) transcriptional activation induced by duplex RNAs in hepatocellular carcinoma (HCC) cell line BEL-7402., Methods: Cells were treated with dsRNAs complementary to promoter sequences of p21(WAF1/CIP1). Quantitative polymerase chain reaction (qPCR) and Western blot were employed to detect the expression of p21. At various timepoints post-transfection, cell viability assay and apoptosis analysis were used to determine the effect of RNA activation. After transfection Western blot was also performed to detect the expression of Bcl-xL, cleaved caspase-3, cleaved caspase-9 and cleaved PARP., Results: DsP21-322 transfection significantly inhibited cell viability. And, at Day 5, dsP21-322 inhibited cell growth by 65.84% versus control. Flow cytometry revealed that dsP21-322 caused a significant increase of cell apoptosis. The total percent of apoptotic cells (UR+LR) increased to 36.86% versus 11.51% and 14.06% in mocks and controls respectively. Such phenomena correlated with a decrease of anti-apoptotic protein Bcl-xL and an increase of cleaved caspase-3, cleaved caspase-9 and cleaved PARP., Conclusion: Activation of p21 gene expression by saRNA may offer therapeutic benefits for HCC and other cancers.

Objective: To investigate the effects of adenoviral vector-mediated over-expression of Wnt3 on the apoptosis of hepatic progenitor cells in vitro., Methods: Hepatic progenitor cells transfected with Ad-GFP-Wnt3 vector or the control vector Ad-GFP were examined for cell apoptosis under fluorescence microscopy with Hoechst 33342 staining, and the proportion of apoptotic cells were determined by flow cytometric analysis with Annexin-PE/7-ADD staining. The mRNA and protein expressions of Bax, Bcl-2 and Bcl-xl in the cells were detected by real-time PCR and Western blotting, respectively., Results: Real-time PCR and Western blotting showed a high expression of Wnt3 in Ad-GFP-Wnt3-transfected hepatic progenitor cells, which exhibited significantly decreased cell apoptosis as compared with the control group. The expressions of Bcl-2 and Bcl-xl mRNA and proteins increased significantly while Bax expression decreased obviously in Ad-GFP-Wnt3-transfected cells (P<0.05)., Conclusions: Adenoviral vector-mediated over-expression of Wnt3 can suppress apoptosis of hepatic progenitor cells possibly through the Bcl-2 pathway.

Objective: To detect the expression of prostate cancer antigen-1 (PCA-1) in prostate cancer, and to analyze the effects of downregulation of PCA-1 expression on malignant biological behavior of prostate cancer LNCaP cells, and to explore their possible molecular mechanisms., Methods: PCA-1-siRNA and control siRNA were transfected into LNCaP cells with lipofectamine 2000. The cell cycle, proliferation and migration were determined by methyl thiazolyl tetrazolium (MTT) assay, flow cytometry and Transwell chambers, respectively. Western blotting was used to detect the expression of cyclin E, matrix metallopeptidase 9 (MMP-9) and p21. Immunohistochemistry was used to detect the expression of PCA-1 protein in 126 cases of prostate cancer and 88 cases of benign prostatic hyperplasia (BPH)., Results: The positive rate of PCA-1 expression was 77.8% (98/126) in prostate cancer, and 10.2% (9/88) in BPH, and its expression was not significantly related to age, prostate specific antigen (PSA), Eastern Cooperative Oncology Group (ECOG) score (P > 0.05), and was associated with Gleason score, TNM staging and bone metastasis (P < 0.05). Downregulation of PCA-1 expression inhibited cell proliferation, arrested cell cycle at S phase and decreased cell migration of LNCaP cells. The downregulation of PCA-1 expression decreased the expression of Bcl-xl, cyclin E and MMP-9 proteins, but increased the expression of p21 proteins., Conclusions: PCA-1 may play an important role in the development of prostate cancer. The downregulation of PCA-1 expression can lead to changes in the proliferation, cell cycle and migration of prostate cancer LNCaP cells, and these effects may be associated with the decrease of Bcl-xl, cyclin E and MMP-9 proteins and increase of p21 protein.

This study was aimed to investigate the effect of Btk inhibitor PCI-32765 and the proteasome inhibitor bortezomib on Raji and Ramos cell proliferation, apoptosis, and its mechanisms. Raji and Ramos cells were treated with PCI-32765 and bortezomib alone and/or their combination. The cell proliferation and apoptosis were detected by CCK-8 and flow cytometry respectively, the expression level of Btk, NFκB, c-IAP1, Bcl-xL and caspase-3 protein were measured by Western blot. The results indicated that: (1) after Raji and Ramos cells were treated with PCI-32765 (0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0 µmol/L) alone and bortezomib (10, 20, 30, 40, 50, 60, 80 nmol/L) alone and their combination for 48 h, the cell proliferation and vitality were inhibited in a dose-dependent manner and both had synergistic effect; (2) Raji and Ramos cells were treated with PCI-32765 (2.0 µmol/L) and bortezomib (20 nmol/L) alone and their combination for 8, 12, 24, 36, 48 and 72 h, the cell proliferation and vitality were inhibited in a time-dependent manner, the two drugs displayed a synergistic effects; (3) the Raji and Ramos cells were treated with PCI-32765 (2.0 µmol/L) and bortezomib (20 nmol/L) alone and their combination for 48 h, all these treatments could induce significant apoptosis of Raji and Ramos cells.In Raji cell experiment, the cell apoptosis rate in the control group, PCI-32765 group, bortezomib group and PCI-32765 and bortezomib combination group were 10.34 ± 0.53%, 24.26 ± 0.91%, 43.66 ± 1.08% and 74.06 ± 0.72% respectively, and the differences was statistically significant among the different groups (P < 0.05). In Ramos cell experiment, the cell apoptosis rate in the control group, PCI-32765 group, bortezomib group and PCI-32765 and bortezomib combination group are 15.16 ± 1.49%, 71.36 ± 0.82%, 75.32 ± 2.36% and 84.30 ± 0.91% respectively, the differences was statistically significant among the different groups (P < 0.05); (4) PCI-32765 and bortezomib could inhibit the expression level of intracellular Btk, NFκB, Bcl-xl and c-IAP1 proteins, but up-regulate the expression level of caspase-3. It is concluded that PCI-32765 and bortezomib can synergistically inhibit the proliferation and induce apoptosis of Raji and Ramos cells, the mechanism may be associated with inhibition of Btk and NFκB activity, down-regulation of anti-apoptotic proteins expression, such as Bcl-xl and c-IAP1, and increase of caspase-3 expression.

Objective of this study was to detect the expression of Survivin in acute myeloid leukemia (AML) and investigate the relationship of its expression levels with clinical variates and its correlations with BCL-2 ,Bcl-xL and MCL-1. The expression of Survivin, BCL-2, Bcl-xL and MCL-1 were measured by immunohistochemistry in bone marrow biopsy of 63 newly diagnosed AML patients, and the relationship between its expression level and clinical parameters (age, sex, WBC count, diagnosis, prognosis), especially fusion protein AML1/ETO was investigated. The results showed that the expression level of Survivin in newly diagnosed AML patients was higher than that of normal controls (P < 0.01), the expression levels of Survivin did not correlate with age, sex, and WBC counts of patients and so on. There was no significant difference of Survivin expression between different NCCN prognosis groups, either between patients with AML1/ETO or FLT3-ITD mutation and the other patients. Survivin positive patients were got to have lower CR rate but higher relapse rate, however that was not significant in statistics. Indeed, the cumulative survivin rate of Survivin positive patients were lower than that of Survivin negative patients (P = 0.04). Finally, positive correlation between Survivin and MCL-1 was also observed (r = 0.639, P = 0.000). It is concluded that overexpression of Survivin are closely related with occurrence and development of acute leukemia, and may be used as an indicator of prognosis evaluation. In addition, Survivin and MCL-1 may have a relationship of cooperation or interaction.

Objective: To investigate the inhibitory effect of NF-kappaB p65siRNA on human laryngeal carcinoma xenograft model in nude mice., Method: Human laryngeal carcinoma cell line Hep-2 was seeded in the subcutaneous layer of 15 nude mice to build laryngeal carcinoma xenograft model. Then they were randomly divided into three groups. NF-kappaB p65siRNA was given in siRNA group and FAM-Control siRNA was given in negative control group while phosphoric-buffered saline (PBS) was used in normal control group for 3 weeks. Tumor size and body weight of the mice were measured. TUNEL method and immunohistochemical S-P method were used for detecting the expression of NF-kappaB p65 and Bcl-xL protein., Result: The volume of tumors in siRNA group was reduced and the average weight of tumors in siRNA group was lower than the other two groups (P < 0.05). In siRNA group, the expression of NF-kappaB p65 and Bcl-xL protein was down-regulated and the apoptotic rate was increased obviously as compared with the negative control group and the normal control group., Conclusion: NF-kappaB p65siRNA can significantly inhibit the expression of NF-kappaB p65 and the growth of human laryngeal carcinoma xenograft in nude mice. Its mechanism may be related to inducing the apoptosis in tumor cells by down-regulating the expression of Bcl-xL protein.

Objective: To clarify the effects and mechanisms of homoharringtonine (HHT) monomer therapy or combination therapy with arsenic trioxide (ATO) on human multiple myeloma (MM) cell line RPMI 8226 in in vitro researches., Methods: Effects of HHT, ATO, and HHT combined ATO on the growth of MM cell line RPMI 8226 were detected using MTT assay. The morphological changes of cell apoptosis were detected by Hoechst staining. The early apoptosis rate was detected using flow cytometry. Expressions of Caspase-3, Caspase-9, poly-ADP-ribose polymerase (PARP), Bcl-2, Mcl-1, Bcl-xl, and AKT protein were detected by Western blot., Results: HHT and ATO inhibited the proliferation of RPM1 8226 cell line in a time- and dose-dependent manner (P < 0.05). Synergistic effects was shown in the combination group (Cl < 1). HHT and ATO induced the apoptosis of RPMI 8226 in a dose-dependent manner with typical morphological changes of apoptosis and higher early stage apoptosis rate. The enhancement in apoptotic induction was seen when two agents were combined. HHT activated expressions of Caspase-3 and PARP in a dose dependent manner at 24 h. HHT at 40 ng/mL and ATO at 8.5 micromol/L could significantly activate expressions of Caspase-3 and Caspase-9, and down-regulate expressions of anti-apoptotic proteins Bcl-xl and Mcl-1. In addition, the combination therapy of HHT at 40 ng/mL and ATO at 8.5 micromol/L inhibited phosphorylation of AKT in a time-dependent manner., Conclusion: HTT, ATO, and combination therapy of HHT and ATO induced the apoptosis of RPMI 8226 cell line possibly through activating Caspase pathways, regulating expressions of Bcl-2 families, and inhibiting phosphorylation of AKT.

Objective: To investigate the function of Parkinson's disease (PD)-related protein Pink1 in autophagy., Methods: Pink1, autophagy inhibitor Bcl-XL and autophagy induced-factor Beclin1 were amplified with RT-PCR and contructed into pcDNA3. 1 (+) vector. The stable HEK293 cell line of Pink1 expression was established through the lentivirus system. Pink1, Bcl-XL and Beclin1 were tansfected into the HEK293 Cell Line. Co-Immunoprecipitation and Western blot were performed to determine the interaction between Pink1 and Bcl-XL, the effect of Pink1 on the interaction between Bcl-XL and Beclin1, and the function of Pink1 in autophagy., Results: There was a new interaction between Pink1 and antophagy inhibitor Bcl-XL. Overexpressed Pink1 promoted the dissociation of autophagy induced-factor Beclin1 from Bcl-XL. Overexpressed Pink1 increased the autophagic protein LC3 II/LC3 I., Conclusion: Pakinson's disease related protein Pink1 promotes the dissociation of autophagy induced-factor Beclin1 from antophagy inhibitor Bcl-XL and promotes autophagy.

Objective: To discuss the mechanism of rectal cancer apoptosis induced by preoperative chemoradiotherapy and evaluate its effect by detection of apoptosis related proteins in locally advanced colorectal cancer patients who had received preoperative chemoradiation., Methods: To detect Bcl-XL and Bax expression in rectal cancer before and after chemoradiotherapy by EnVision method, combined with patients clinical and pathological index, statistically analysis and evaluation their relationship and clinical significance., Results: Patients with or without tumor shrinkage after preoperative chemoradiotherapy was 13 cases and 21 cases. While the positive rate of Bcl-XL in rectal cancer before and after chemoradiotherapy were 58.8% (20/34) and 52.9% (18/34), respectively. There were significant difference between Bcl-XL change before and after chemoradiation with tumor size, tumor cells shrinkage and operation pattern. The positive rate of Bax in rectal cancer before and after chemoradiotherapy were 32.4% (11/34) and 44.1% (15/34), respectively. There were no significant difference between Bax change before and after chemoradiotherapy with tumor cells shrinkage. There were statistically significant difference between Bax ratio (χ(2) = 9.607, P = 0.048) before and after chemoradiation while there were no significant difference between Bcl-XL/Bax ratio before and after chemoradiation with tumor shrinkage. According to layered analysis with preoperative therapy, there were statistically significant difference (χ(2) = 13.964, P = 0.007) between Bcl-XL change with operation pattern while the same of significant difference between Bax change with tumor infiltration and tumor shrinkage (χ(2) = 10.806 and 10.455, both P < 0.05)., Conclusions: Preoperative chemoradiation can influence rectal cancer cell's apoptosis and treatment effect by changing Bcl-XL and Bax expression. Bcl-XL downregulation and Bax upregulation have shown important function in colorectal cancer cell apoptosis which induced by preoperative chemoradiation, it can also improve the effection of chemoradiation in rectal cancer.

Objective: To investigate the apoptosis effect of diffuse large B-cell lymphoma cell line (DLBCL) SUDHL-4 induced by IL-21 and its related mechanism., Methods: SUDHL-4 cells were treated with IL-21 at different concentration (1000 ng/ml, 100 ng/ml, 10 ng/ml, 1 ng/ml) for 24 h, 48 h, 72 h, respectively. The inhibitory rate of cell proliferation was detected by CCK-8 assay. The cell growth curves were drawn and half inhibitory concentration (IC(50)) values were calculated. The cell apoptosis were detected by flow cytometry (FCM), the expression of the caspase-9, caspase-3, cleaved caspase-3, Bcl-2, Bcl-XL, Bid, Bax and c-myc protein in SUDHL-4 cells treated with IL-21 by western blot, the mRNA expression of Bcl-2, Bcl-XL, Bid, Bax, c-myc by Survivin gene with RT-PCR., Results: IL-21 markedly inhibited SUDHL-4 cell growth in a time- and dose-dependent manner. The 48 hIC(50) was 140.9ng/ml; The FCM showed that the apoptosis proportion of SUDHL-4 cells treated with 100 ng/ml of IL-21 apoptosis (AnnexinV-FITC(+) positive cells) gradually increased (48 h: 19.7 ± 2.3%). The protein expression of caspase-9, caspase-3, Bcl-2 and Bcl-XL decreased in a time-dependent manner. The Bax and c-myc protein markedly increased, but the Bid protein level did not change. IL-21 up regulated c-myc and Bax gene expression, however down regulated Bcl-2 and BCL-XL gene expression, but the gene expression of Bid and Survivin hadn't been changed significantly., Conclusions: IL-21 can inhibit proliferation and induce apoptosis of SUDHL-4 cell. The mechanism may involve in endogenous mitochondrial pathway mediated by the c-myc and the Bcl-2 genes.

Objective: To observe the effect of sodium valproate (VPA) on the proliferation and apoptosis of human lung carcinoma SPC-A1 cells and the underlying mechanism., Methods: The effect of VPA on the proliferation of SPC-A1 cells was evaluated by MTT assay and clone formation assay. Flow cytometry was used to analyze the apoptosis of the cells exposed to VPA. The changes in the expressions of Bcl-xl, Bcl-2, Mcl-1, caspase-9, and caspase-3 in the exposed cells were detected by Western blotting., Results: Incubation with VPA for 48 h resulted in a significant inhibition of SPC-A1 cell proliferation, with a IC(50) of 1.8 mmol/L. VPA treatment also inhibited cell colony formation and induced obvious cell apoptosis. Exposure to 8 mmol/L VPA for 48 h caused a percentage of early apoptotic cells of 60.44%. VPA treatment at different concentrations for 48 h obviously lowered the protein levels of Bcl-xl, Bcl-2, and Mcl-1 and induced caspase-9 and caspase-3 activation in SPC-A1 cells., Conclusion: VPA can inhibit the proliferation of SPC-A1 cells by triggering mitochondrion-dependent apoptosis.

This study was aimed to investigate the inhibitory effects of sporoderm-broken ganoderma lucidum spores (GLS) on transplanted lymphoma in nude mice and its mechanism. The models of the subcutaneously transplanting tumor were established by N-methylnitrosourea-induced thymus T-cell lymphoma. The cellular apoptosis of tumor tissues in nude mice were observed by transmission electron microscopy (TEM), the mRNA expression of Bax, Bcl-2, Bcl-xl was determined by RT-PCR. The results indicated that the GLS had a certain anti-tumor effect, and the inhibitory rate was 45.8 with the dose of 4 g/kg (ig, once daily for 21 d). Apoptotic cells in lymphoma cells treated with GLS were observed by TEM. RT-PCR analysis showed that the expression of Bax was up-regulated, Bcl-2 and Bcl-xl were down-regulated in T-cell lymphoma cells. It is concluded that GLS can inhibit proliferation of lymphoma cells and induce the lymphoma cell apoptosis, the mechanism of which may be related with up-regulating the expression of Bax and down-regulating the expression of Bcl-2 and Bcl-xl.

Objective: To investigate the anti-tumor effect of adenovirus-mediated Bcl-XL shRNA on colon cancer cells in vitro., Methods: A recombinant Bcl-xl adenovirus was constructed, amplified, and purified. The effect on mRNA expression of Bcl-XL was assessed by RT-PCR, and the effect on apoptosis-induction of colon cancer(Lovo cell line) in vitro was assessed by MTT assay and cell clonogenic assay., Results: RT-PCR showed that Ad/Bcl-XL shRNA significantly down-regulated the mRNA expression of Bcl-XL in Lovo cells. Ad/Bcl-XL shRNA suppressed the proliferation of Lovo cells in a dose-dependent as well as a time-dependent manner compared with Ad/GFP (P<0.05). Treatment with Ad/Bcl-XL shRNA dramatically suppressed the colony formation of Lovo cells in a dose-dependent manner (P<0.05). Ad/Bcl-XL shRNA showed no effect on normal human fibroblast., Conclusion: Ad/Bcl-XL shRNA exhibits cytotoxic effect on Lovo cells and may have the potential value in the treatment of colon cancer.

Objective: To investigate the effect of Angiotensin(1-7) [Ang(1-7)] on left ventricular dysfunction and myocardial apoptosis on rat model of adriamycin-induced dilated cardiomyopathy (ADR-DCM)., Methods: Weight-matched adult male Wistar rats were randomly divided into 3 groups: (1) the ADR-DCM group (n = 25), in which 2.5 mg/kg of ADR was weekly intravenously injected for 10 weeks. (2) Ang(1-7) group (n = 25), in which ADR rats were simultaneously treated with angiotensin-(1-7) (24 µg×kg(-1)×h(-1), ip.) for 12 weeks. (3) normal control group (n = 10). Hemodynamics and echocardiography examination were performed at 12 weeks. The malondialdehyde (MDA) was measured by TBA methods. The plasma concentration of AngII was determined by immunoradiometric assay. The pathological change was analyzed by histological hematoxylin-eosin staining. Myocardial apoptosis was assessed by TUNEL method. The protein expression of pro-apoptotic protein caspase-3, Bax and anti-apoptotic protein Bcl-xl in cardiomyocytes were detected by Western blot., Results: Mortality was significantly lower in Ang(1-7) group than in ADR-DCM group (16% vs. 40%, P < 0.01). Compared to the control group, left ventricular end-diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD) and left ventricular end-diastolic pressure (LVEDP) were significantly increased in ADR-DCM group (all P < 0.01) while fractional shorting (FS), +dp/dtmax and -dp/dtmax were significantly reduced in ADR-DCM group (all P < 0.01). LVEDD, LVESD and LVEDP were significantly reduced while FS, +dp/dtmax and -dp/dtmax were significantly higher in Ang(1-7) group compared to the ADR-DCM group, but still higher than the control group (all P < 0.01). The concentrations of AngII and MDA were higher in the ADR-DCM group than in the control group (P < 0.01), which were significantly reduced by Ang(1-7) treatment (P < 0.01). The TUNEL-positive cells and apoptosis index, the expression of pro-apoptotic protein caspase-3 and Bax were significantly higher while the expression of anti-apoptotic protein Bcl-xl was significantly lower in the ADR-DCM group than in the control group (all P < 0.01) which could all be partially reversed by Ang(1-7) treatment (all P < 0.01)., Conclusion: Ang(1-7) could significantly attenuate left ventricular dysfunction and myocardial apoptosis in this model by downregulating pro-apoptotic protein caspase-3 and Bax and upregulating anti-apoptotic protein Bcl-xl expression.

Objective: To investigate the synergistical killing effect of docetaxel combined with ABT-737 on human prostate cancer cell line PC-3 by inducing apoptosis and further to determine the mechanism underlying such effect., Methods: PC-3 cells were treated with various concentrations of docetaxel or (and) ABT-737. Cell viability was determined using MTT assay. Apoptosis was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with 4',6-diamidino-2-phenylindole (DAPI). Cellular DNA was stained with propidium iodide and flow cytometric analysis was performed to analyze the cell cycle distribution. Bcl-2, Bax, Bcl-xL and Mcl-1 protein changes were detected by Western blot. The activity of caspase-3 was measured using a colorimetric assay., Results: Docetaxel (20 nmol/L) combination with ABT-737 (400 nmol/L) for 48 hours, the cell viability was decreased to 19.7% ± 3.2% to compare with 44.2% ± 4.4% (t = 4.45) of docetaxel and 93.2% ± 1.8% of ABT-737 separately and there was a synergistic effect between the two drugs (CI = 0.8). Apoptosis rate of the combination group was higher than other two drugs. Docetaxel increased the cell number arrested in G(2)/M phase compared with control group (P < 0.05), but the combination treatment resulted in a significant arrest in the G(0)/G(1) phase. The combination treatment could significantly reduced the Bcl-2, Bcl-xL and Mcl-1 expression (F = 369.53, 57.89 and 32.77, all P < 0.05) and enhanced the activity of caspase-3 (419.7% ± 15.6%) (F = 207.33, P < 0.05)., Conclusions: The combination of ABT-737 with docetaxel can synergistically inhibit the proliferation of PC-3 cells through inducing apoptosis, which may be associated with cell cycle arrest, down-regulation of Bcl-2, Bcl-xL and Mcl-1 expression and activation of caspase-3.

Objective: To study the effect of ABT-737 combined with cisplatin on apoptosis of breast cancer cell line T47D cells., Methods: T47D cells cultured in vitro was used for this experiment. Cell proliferation was measured by MTT assay. The expression of apoptosis-related protein was determined by Western blot. Morphological changes of apoptotic cells were observed by fluorescence microscopy. The apoptosis rate was examined by flow cytometry., Results: The MTT assay showed that ABT-737 significantly decreased the IC(50) of cisplatin in T47D cells [(26.00 ± 1.41) µmol/L of single cisplatin vs. (13.00 ± 1.11) µmol/L of combination (ABT-737 + cisplatin)]. As a single agent, ABT-737 did not inhibit the proliferation of T47D cells, but enhanced the inhibitory effect of cisplatin in a dose-dependent manner. The detection of the cleavage of PARP showed that ABT-737 lowered the doses of cisplatin to induce apoptosis and shortened the induction time of apoptosis in T47D cells. Compared with the single use of cisplatin, the combination of ABT-737 and cisplatin accelerated the cleavage of PARP and caspase3, but did not alter the expression levels of Bcl-2, Bcl-X(L), and Bax. Both flow cytometry and fluorescence microscopy showed that ABT-737 combined with cisplatin significantly increased the apoptosis induction in T47D cells (2.3% ± 0.1 % in the control, 30.0% ± 0.8% in the cisplatin alone, and 49.0% ± 0.5% in the cisplatin + ABT-737 groups, P < 0.05)., Conclusion: The Bcl-2 inhibitor ABT-737 can significantly enhance cisplatin-induced apoptosis in human breast cancer T47D cells in vitro.

Objective: To evaluate the effects of valsartan and carnitine on cardiomyocyte Calpain-1 and Bcl-xl expressions of dogs with chronic alcohol intake-induced cardiomyopathy., Methods: Dogs were randomly assigned into 4 groups (n = 7 each): (1) alcohol fed (free access to 5%, 1(st) week; 10% 2(nd) week; 500 ml 25% bolus plus free access to 5% from 3 to 24 weeks, A); (2) alcohol + valsartan (5 mg×kg(-1)×d(-1), B); (3) alcohol + carnitine (300 mg×kg(-1)×d(-1), C); (4) Control (D). After six months, all animals were assessed for left ventricular (LV) function by echocardiography. The Bad and Bcl-xl protein expressions were evaluated by immunohistochemistry. The expression of Calpain-1 protein was determined with Western blot. Myocardial morphology was quantified on HE stained slices and under electron microscopy. The terminal deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) was performed for apoptosis analysis., Results: Compared with group D, LVEDD and LVESD were significantly increased while EF and FS significantly decreased in group A. In alcohol fed group, expressions of Bad and Calpain-1 protein were significantly increased while Bcl-xl protein expression was downregulated, all changes could be significantly attenuated by intervention with valsartan and carnitine (all P < 0.05)., Conclusion: These data suggest that alcohol could promote cardiac myocyte apoptosis, reduce cardiac function and aggravate myocardial remodeling which valsartan and carnitine could reduce alcoholic cardiomyopathy by downregulating Calpain-1 and Bad protein expression and upregulating expression of Bcl-xl protein.

Objective: To investigate the different efficacy of Yinchenhao Decoction (YCHD), a compound traditional Chinese herbal medicine, for liver cirrhosis induced by dimethylnitrosamine (DMN) or carbon tetrachloride (CCl(4)) in rats., Methods: To induce liver fibrosis, 0.5% DMN solution (2mL/kg body weight, i.p.) was given three consecutive days a week to male Wistar rats for 4 weeks. Cirrhotic rats were randomly divided into DMN group, YCHD group, Xiaochaihu decoction group by the end of the fourth week to accomplish a 2-week recipe treatment course. In CCl(4)-induced liver fibrosis model, 50% CCl(4)-olive solution was injected subcutaneously to rats at a dose of 2 mL/kg body weight twice a week to duplicate rat cirrhosis model. After 8 weeks, rats were divided into CCl(4) group, CCl(4) plus YCHD group and Xiaochaihu decoction group. For the YCHD group, YCHD was administered intragastrically once a day for 4 weeks. For DMN or CCl(4) model, by the end of 6 or 12 weeks respectively, rats were sacrificed for sampling to detect liver function, hepatic histological changes, hydroxyproline (Hyp) content and apoptosis-related gene expressions., Results: In DMN liver fibrosis model, hepatic fibrosis was obvious at week 2 and cirrhosis was evident at week 4 in DMN-treated rats. Compared to 6-week DMN group, hepatic pathological changes and liver function were improved significantly and content of Hyp decreased remarkably in YCHD group. In CCl(4)-induced liver fibrosis model, hepatic fibrosis was obvious at 8 weeks and cirrhosis was evident at 12 weeks in CCl(4)-treated rats. Compared to 12-week CCl(4) group, hepatic pathological changes and liver function were not obviously improvement in YCHD group. The results of gene chip showed that YCHD significantly decreased Fas, Bax and caspase-3 gene expressions, and increased Bcl-xL gene expression in the liver of DMN model. However, in the model induced by CCl(4), YCHD did not inhibit hepatocyte apoptosis induced by CCl(4), but increased tyrosine kinase receptor gene expression by 4.8 times., Conclusion: YCHD exerts more significant therapeutic effects on DMN-induced than CCl(4)-induced cirrhosis in rats in Hyp content and pathological change in liver tissue.

This paper was aimed to investigate the biodistribution and ability of free 131-bcl-2/bcl-xl ASON (FA) and anionic long circulation liposomes encapsulated with 131I-bcl-2/bcl-xlASON (NA), in tumor-bearing rats, to image breast cancer. We investigated the tissue distribution of NA in virgin female Sprague-Dawley (SD) rats with n-methyl nitrosourea (MNU)-induced breast cancers in situ. The percentage of the injected dose per gram (%ID/g) was calculated, with the maximum ratios of tumor to blood and tumor to muscle, after injections of NA and FA for 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, 12 h and 24 h, respectively. The ability of NA to image breast cancer in tumor-bearing rats was determined using emission computed tomography (ECT). Seventy percent (90/130) SD rats in the study developed mammary tumors after MNU injection with the average latency (NA) (96 +/- 1.2)days. The %ID/g of NA in breast cancer tissue, tumor bearing rats in liver and spleen tumor tissues after 10 hours were (6.23 +/- 0.23) %ID/g, (12.00 +/- 0.26) %ID/g and (18.25 +/- 1.33)% ID/g, respectively. The ratios of tumor to blood 6.29 +/- 0.76 and tumor to muscle 10.55 +/- 0.68 in tumor bearing rats slowly maximized at 10 h post injection of NA, most probably due to the enhanced permeability and retention effect. Hence in radionuclide antisense scintigraphy, the breast cancer in rat was clearly displayed at 10h after iv administration of NA-D. However, tumors were not visualized in rats with the iv injection of NS and NN even at the delayed time. Due to the inhibition of rapid uptake of NA by the reticulo-endothelial system, NA displays valuable pharmacologic properties characterized by the enhanced accumulation in tumor.

Objective: To indentify the relation between hepatic cells apoptosis and the lesion of liver tissue in acute toxic lethal hepatitis., Methods: 60 Wistar mice were randomly divided into normal control, model group and treatment group. Normal control and model group were pretreated by portal vein injection of normal saline, the treatment group was pretreated by portal vein injection of BCL-X1 adenoviruses. The mice of model group and treatment group were received an injection of D-galn and LPS to establish fulminant hepatic failure models 7 days after pretrement. To observe BCL-X1 expression, serum ALT, AST, hepatocyte apoptosis rate, and mortality rate of the three groups., Results: The BCL-X1 expression was higher in treatment group than in model group; 6 hours after fulminant hepatic failure models were established,the serum ALT, AST level of treatment group was lower than model group;The hepatocyte apoptosis rate of treatment group was lower than model group. The death rate of treatment group was lower than model group., Conclusion: In fulminant mice hepatic failure models, the hepatocyte apoptosis rate has a positive correlation with death rate, the overexpression of BCL-X1 can decrease the hepatocyte apoptosis rate and the death rate.

Objective: To observe the inhibitive effects of Plastrum testudinis Extracts (PTE) on 6-Hydroxydopamine (6-OHDA) induced PC12 cells apoptosis and explore its mechanism., Methods: PC12 apoptosis model was established by serum starvation and damaged for 24 hours. The cells were randomly divided into four groups:control group, 6-OHDA group, PTE 3, 30 microg/mL group. Cell optical density was determined by MTT; Ratio of cell apoptosis was examined by Annexin V/PI double stain flow cytometry (FCM), and Western blot was applied to detect the BCL-X/L expression., Results: MTT and FCM analysis demonstrated that PTE can elevate PC12 cells viability and reduce their apoptotic ratio in a dose dependent manner. Western blot showed that PTE promoted the expression of BCL-X/L., Conclusion: PTE can inhibit the apoptosis of PC12 induced by 6-OHDA in a dose dependent manner, and its mechanism maybe associated partially with up-regulating BCL-X/L signaling pathway.

Objective: To study the effects of siltuximab on the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (Stat3) signaling pathway in ovarian epithelial carcinoma., Methods: (1) Expressions of IL-6 in ovarian cancer patient specimens were assessed by immunohistochemistry. (2) Expression of phosphorylation Stat3 (pStat3) protein in siltuximab and IL-6 treated SKOV3 cell lines was determined by western blot, and expression levels of Stat3-induced bcl-XL, MCL-1, survivin, in siltuximab treated SKOV3/TR and CAOV3/TR cells lines were also determined by western blot. (3) Real-time image analysis was used to study the nuclear translocation of pEGFP-Stat3 fusion protein in ovarian cancer cell line SKOV3-pEGFP-Stat3 treated with siltuximab and IL-6. (4) Paclitaxel sensitivity in siltuximab treated SKOV3/TR and CAOV3/TR cell lines were assessed using the methyl thiazolyl tetrazolium (MTT). The 50% inhibiting concentration (IC(50)) was defined as the paclitaxel concentration required to decrease the A(490) value to 50%., Results: (1) There were significantly difference in IL-6 staining density and the positive rate of IL-6 protein stained among the metastatic, and drug-resistant recurrent tumors, and matched primary tumors [69% (18/26)] vs. 77% (20/26) vs. 23% (6/26), P < 0.05]. (2)A clear increase in Stat3 phosphorylation levels was observed in the IL-6-treated SKOV3 cell lines as compared to the SKOV3 cell lines. When the IL-6-treated SKOV3 cells were incubated with siltuximab with a range of concentrations of 0.001, 0.01, 0.1, 1.0 and 10 µg/ml, there were trends toward reduced pStat3 expression in the treated cell lines. Compared without treatment with siltuximab, the expression of the anti-apoptotic proteins MCL-1, bcl-XL and survivin in SKOV3/TR and CAOV3/TR cell lines were significantly decreased after treated with siltuximab. (3) In resting cells, the majority of pEGFP-Stat3 was cytoplasmic until the addition of human IL-6, which promptly induced the translocation of fluorescent Stat3 molecules to the nucleus. Exposure of cells to siltuximab with a range of concentrations of 0.001, 0.01, 0.1, 1.0 and 10 µg/ml, followed by an incubation in IL-6 significantly reduced pEGFP-Stat3 nucleocytoplasmic translocation. (4) MTT cytotoxicity assay demonstrated that siltuximab increased paclitaxel-induced cell death and partially overcame paclitaxel resistance. Treated with siltuximab (1 and 10 µg/ml), the paclitaxel IC(50) value of siltuximab in SKOV3/TR (0.49, 0.19 µg/ml) and CAOV3/TR (0.0010, 0.0008 µg/ml) cells were significantly lower than those in untreated cells (0.71, 0.0021 µg/ml; all P < 0.05)., Conclusions: These results demonstrated that siltuximab effectively block the IL-6 signaling pathways, which. Blockage of IL-6 signaling may provide benefits for the treatment of ovarian cancer.

Objective: To study the effect of pGCsi-H1-APRIL on the growth of human colorectal cancer cells in transplated tumor in nude mice and to improve the effect of APRIL on proliferation and apoptosis of colorectal cancer (CRC)., Methods: Human CRC model was established in nude mice, and the nude mice were treated with APRIL siRNA twice per week for 2 weeks. APRIL mRNA expression was surveyed by PCR and APRIL protein expression was detected by immunohistochemistry. The expression of PCNA protein was detected by ELISA. The expression of bcl-2 and bcl-xl was assessed by immunohistochemical staining, and TUNEL staining was used to detect apoptosis., Results: The expression of APRIL mRNA in the APRIL siRNA group was (0.13 ± 0.05) × 10(-3), significantly lower than that in the vector group (0.95 ± 0.04) × 10(-3) and the PBS group (0.96 ± 0.05) × 10(-3). The expression of APRIL protein in the APRIL siRNA group was (87.5 ± 5.0)% lower than that in the vector and PBS groups (P < 0.05). APRIL siRNA significantly suppressed the growth of SW480 tumor: the IR (inhibitory rate) of APRIL siRNA group was (60.7 ± 1.5)% (P < 0.05). The expression of PCNA in APRIL siRNA group was (176.8 ± 18.1) ng/ml, was (56.5 ± 2.0)% lower than that of PBS group (328.4 ± 22.8) ng/ml. Furthermore, the expressions of anti-apoptosis proteins bcl-2 and bcl-xl of APRIL siRNA group were (82.6 ± 4.5)% and (79.2 ± 3.5)% lower than those of the PBS group. The apoptotic rate of the APRIL siRNA group was 40.1% ± 2.5%, significantly higher than that in the vector group (2.5 ± 0.1)% and PBS group (2.5 ± 0.2)% (P < 0.05)., Conclusion: APRIL siRNA may significantly suppress the growth and promote apoptosis in transplanted tumor of human colorectal cancer in nude mice. APRIL may become a candidate gene of gene therapy of human colorectal cancer.

This study was aimed to explore the effects of STI571 alone or with As₂O₃ on proliferation, apoptosis and caspase 3, bcl-xL mRNA expression of K562 cells, and the molecular mechanism of As₂O₃ enhancing the anti-leukemia effect of STI571 so as to provide the scientific basis for clinical treatment of chronic myeloid leukemia. The effect of drugs on proliferation of K562 cells was assayed by MTT method, the apoptosis rate of K562 cells was detected by flow cytometry with Annexin V/PI double staining, the caspase 3, bcl-xL mRNA expressions of K562 cells were determined by real time quantitative PCR. The results showed that STI571 alone or with As₂O₃ both could inhibit the proliferation of K562 cells. OD value in test groups reduced along with prolonging of action times, the OD values between different time points were significantly different (p < 0.05), furthermore the OD values at 72 hours in test groups were lowest, while as compared with control group, OD values at same time points in test groups all gradually decreased, among which decrease of OD value in test 5 group was most significant. The flow cytometric detection indicated that along with time prolonging, the apoptotic rate in control group not obviously changed, but the apoptotic rate in test groups gradually increased, the difference between time points was significant (p < 0.05), moreover apoptotic rate increased most obviously at 72 hours, while as compared with control group, apoptotic rate at same time points in test groups was gradually enhanced (p < 0.05), among which the apoptotic rate in test 5 group was highest. The real time qPCR assay revealed that as compared with control group, the bcl-xL mRNA expression in test groups reduced with decrease of 2-ΔΔCT value, furthermore the decrease of expression level in test 3 group was higher than that in test 2 group (p < 0.05), while the caspase 3 mRNA expression in test groups was enhanced with increase of 2-ΔΔCT value, moreover the increase of expression level in test 3 group was higher than that in test 2 group (p < 0.05). It is concluded that the STI571 can inhibit the proliferation of K562 cells, accelerate the apoptosis of K562 cells. The STI571 combined with As₂O₃ can enhance these two effects, increase the expression of caspase-3 mRNA and decrease the expression of bcl-xL mRNA. Therefore, the effect of STI571 combined with As₂O₃ on expression of caspase 3 and bcl-xL mRNA may be one of molecular mechanisms underlying their synergic antileukemia efficacy.

Objective: To explore the protection effects of acupuncture on glaucomatous optic nerve damage and its mechanism., Methods: Experimental glaucoma model was induced by intracameral injection of compound Carbomer solution in rabbits. After 28 days, ocular tension returned to normal by filtration surgery. Twenty rabbits (40 eyes) were randomly divided into a model group, an acupuncture group, a neurotrophy group and a nomal control group, 5 cases in each group. The acupuncture group was treated with acupuncture at bilateral "Qiuhou" (EX-HN 7),"Fengchi" (GB 20) and "Xingjian" (LR 2), twice a day. The neurotrophy group was treated with intramuscular injection of Vitamin B1 (100 mg) and Vitamin B12 (500 microg), once a day, and the other groups with no treatment. The expressions of Bcl-xl and BDNF in rabbits retina were observed after 4 weeks., Results: At the end of the experiment, the positive expression cells of Bcl-xl and BDNF were (31.20 +/- 5.97) per mm2 and (6.3 +/- 1.89) per mm2 in the acupuncture group, being significantly higer than (26.70 +/- 4.32) per mm2 and (4.0 +/- 1.89) per mm2 in the model group (both P<0.05)., Conclusion: Acupuncture can raise the expression of Bcl-xl and BDNF of retina, so as to prevent optic nerve damage caused by intraocular hypertension.

This study was purposed to explore the inhibitory effect of Curcumin on growth of retinoic acid-resistant acute promyelocytic leukemia (APL) cells and its mechanism. The NB4-R1, an APL cell line resistant to retinoic acid, was used as a model. The growth level of NB4-R1 was detected by MTT assay, the morphologic features of cells were observed by light microscopy, the mitochondrial transmembrane potential was determined by flow cytometry, the expressions of apoptosis-related proteins procaspase 3, caspase 3, PARP and BCL-XL were measured by Western blot. The results indicated that the sensitivity of NB4-R1 to Curcumin was consistent with NB4 though NB4-R1 was resistant to retinoic acid, Curcumin displayed inhibitory effect on growth of NB4-R1 in time-and concentration-dependent manners. The morphologic observation showed existence of apoptotic bodies in NB4-R1 cells treated with 20 micromol/L of Curcumin. The flow cytometry indicated that the mitochondrial transmembrane potential in NB4-R1 cells treated with 20 micromol/L of Curcumin obviously decreased. The Western blot detection revealed that expressions of pro-caspase 3 and BCL-XL were down-regulated, expressions of caspase 3 and sheared PAPP were up-regulated in NB4-R1 cells treated with 20 micromol/L of Curcumin. It is concluded that the Curcumin can inhibit the growth and induce the apoptosis of NB4-R1.

Objective: To investigate the protective effects of Sini decoction (SND) on Adriamycin-induced heart failure and its mechanism., Methods: SD rats were randomly divided into three groups:control group,heart failure model group and SND group. ADR was injected in to the rats of model group and SND group by caudal vein. After injection,the rats in SND group were given SND [3.75 g/(kg x d), p.o.]. Three weeks later, protein expressions of Bid and Bcl-xl were detected by immunohistochemistry; mRNA expression ratio of Bcl-xl/Bcl-xs was detected by RT-PCR and apoptosis rate was determinated by flow cytometry., Results: Compared with control group, the protein expression of Bcl-xl and mRNA ratio of Bcl-xl/Bcl-xs obviously decreased,while the protein expression of Bid and apoptosis rate significantly increased in the model group. SND could decrease cell apoptosis, increase the protein expression of Bcl-xl, increase bcl-xl/bcl-xs mRNA ratio and decrease Bid protein expression., Conclusion: Bcl-xl may play an important role in ADR-induced heart failure rats. The mechanism of SND on protecting cardiocyte may be related to apoptosis correlation factor, Bcl-xl and Bid.

Objective: To investigate the expressions of proliferating cell nuclear antigen (PCNA), B cell lymphoma-2 (Bcl-2) and Bcl-associated X (Bax) in the lymphocytes of patients with oral lichen planus (OLP)., Methods: Immunohistochemistry was used for detecting the expressions of PCNA, Bcl-2 and Bax proteins in oral lichen planus tissue and normal oral mucosa (NOM)., Results: The expressions of PCNA and Bcl-2 proteins increased significantly in the lymphocytes of patients with OLP compared with those in NOM. The expressions of PCNA and Bcl-2 proteins showed no significant differences between patients with OLP of general type and erosive type., Conclusion: Increased expression of PCNA and the imbalance of Bcl-2/Bax expressions in the lymphocytes of patients with OLP can be important causes for continuous lymphocyte infiltration in OLP.

Objective: To investigate the suppressive effect of resveratrol on growth of U251 human glioma cells and its correlated mechanism., Method: U251 human glioma cells were treated with resveratrol at various concentrations, MTT assay was used to determine the inhibitory rate of cell proliferation, FCM to detect the cell apoptosis, the expressions of Bcl-2, Bcl-XL, STAT3 and CyclinD1 were analysed by immunohistochemistry and Western blot to examine the expression of Bcl-2, Bcl-XL, STAT3, CyclinD1, Caspase-3 and Bax., Result: After treatment with resveratrol, MTT assay showed the growth of U251 cells was inhibited in dose-dependent and time-dependent manners, apoptosis of cells advanced stage was built up, immunohistochemical staining displayed decreased the expression of Bcl-2, Bcl-XL, STAT3 and CyclinD1 and Western blot showed that resveratrol decreased the expression of Bcl-2, Bcl-XL, STAT3 and CyclinD1, and built up Bax and Caspase-3., Conclusion: It is possible that downregulated the expression of Bcl-2, Bcl-XL, but upregulated Bax and Caspase-3, and the indication was obviously in dose-dependent and time-dependent manners.

This study was aimed to investigate the effects of apogossypolone (ApoG2) on proliferative inhibition and apoptotic induction of multiple myeloma cells and its mechanism. The effects of ApopG2 on cell growth, cell viability, cell cycle and cell apoptosis were determined by Hoechst 33258 staining, DNA ladder formation and subdiploid peak analysis respectively. Cleavage of caspase-3 and caspase-9 was analyzed by colorimetric assay. Expression of BCL-2 and BCL-XL was detected by flow cytometry. The results indicated that the ApoG2 inhibited multiple myeloma cell proliferation in dose-and time-dependent manners, with IC(50) value to both U266 and Wusl cells at 0.1 and 0.2 micromol/L at 48 hours after treatment. ApoG2 effectively inhibited the proliferation of multiple myeloma cells, the IC(50) value in U266 cells and Wusl cells (at 48 hours) were 0.1 micromol/L and 0.2 micromol/L respectively. ApoG2 could induce the apoptosis of cells of myeloma in a time-dependent manner.The typical apoptotic morphological changes were observed under transmission electron microscope, while DNA ladder formation and remarkable peak of subdiploid cells appeared. ApoG2 could arrest the myeloma cells in G(2) phase, increasing from 9.7%(0 micromol/L) to 19.6% (10 micromol/L) in U266 cells and 9.8%(0 micromol/L) to 31.7% (10 micromol/L) in Wusl cells. ApoG2 could induce increase of caspase 9 and caspase 3 activity and down-regulate the expression of BCL-XL in U266 and Wusl cells, as well as the expression of BCL-2 in Wusl cells. It is concluded that ApoG2 has significant effect of antiproliferation and induction of apoptosis on multiple myeloma cells in vitro, ant its mechanisms may involve in down-regulation of BCL-2/BCL-XL and in change of cell cycle.

This study was purposed to explore the apoptotic effect of gambogic acid on Raji cells and the role of death inducer-obliterator 1 (DIO-1) in this process. Annexin V-fluorescein-isothiocyanate/propidium iodide was used to detect apoptosis of Raji cells. Western blot was used to determine the expressions of DIO-1, Bcl-xL, pro-caspase 3 and 2 activated subunits: P17 and P20. The subcellular localization of DIO-1 in untreated and treated Raji cells was checked by immunofluorescence and Hoechst 33258 double staining. The results showed that the Gambogic acid dose-dependently induced the apoptosis of Raji cells, downregulated the expression of Bcl-xL, upregulated the expressions of DIO-1 and pro-caspase 3, induced the cleavage of pro-caspase 3 and DIO nuclear translocation. It is concluded that gambogic acid induces the apoptosis of Raji cells through DIO-1 upregulation, nuclear translocation, Bcl-xL downregulation and caspase 3 activation.

Objective: To investigate the effects of extracts of Solanum lyratum (ESL) on the apoptosis of Human stomach cancer SGC-7901 cells., Methods: Dried whole herbs of Solanum lyratum were extracted by boiling distilled water. SGC-7901 cells were randomly divided into control group, ESL-treated groups (12.5 g/L, 25 g/L, 50 g/L) and the positive control (25 mg/L DDP) group. The growth inhibitory rate was evaluated by MTT assay. Morphological changes of apoptosis were observed with fluorescence microscope. Cell apoptosis rate was determined by flow cytometry. Expressions of bcl-xl, Caspase-9 and bid mRNA were detected by semi-quantitive RT-PCR. The activity of Caspase-3 was detected by Fluorospectrophotometry., Results: Compared with control group, the cell proliferation inhibitory rate and apoptosis rate of human stomach cancer SGC-7901 cells increased obviously (P < 0.05). There were obvious changes of morphology of the SGC-7901 cells as the nuclear shrinkage, chromatin condensation and margination; The expression of bcl-xl mRNA decreased obviously (P < 0.05), the expression of Caspase-9 and bid mRNA increased obviously respectively (P < 0.05), and displayed effect in a dose-dependent manner in the SGC-7901 cells of the ESL-treated groups. The activity of Caspase-3 in the SGC-7901 cells of the ESL-treated groups were higher than that of the control group significantly (P < 0.01)., Conclusion: ESL can induce apoptosis and inhibit proliferation of the human stomach cancer SGC-7901 cells by regulating expression of bcl-xl, Caspase-9 and bid genes and strengthening the activity of Caspase-3.

Objective: To investigate the effects of epigallocatechin-3-gallate (EGCG) on human hepatocellular carcinoma (HCC) cells and mechanism thereof., Method: Human HCC cells of the lines HepG2 and SMMC-7721 were cultured and treated with of EGCG of the concentrations of 6.25, 12.5, 25, 50, 100, 200, and 400 microg/ml respectively for 24 h and 48 h. The cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Trypan blue staining was used to count the cells. Flow cytometry was conducted to detect the cell apoptosis. The protein levels of Bcl-2, an anti-apoptosis factor, and cyclooxygenase-2 (COX-2), an up-regulator of Bcl-2. The activities of caspase-9 and caspase-3 hat promote the apoptosis of HCC cells, were measured using colorimetric method. RT-PCR was used to detect the mRNA expression of COX-2 and Bcl-2 family., Results: The viabilities of the HepG2 and SMMC-7721 cells treated with EGCG of the concentrations of 50 - 400 microg/ml for 48 h reduced to 93.8% +/- 2.8%, 62.3% +/- 5.4%, 33.9% +/- 2.5%, and 17.6% +/- 3.2% respectively, all significantly lower than that of the control group [(100.0% +/- 2.8%), all P < 0.05]; and the viabilities of the SMMC-772 cells treated with EGCG of the concentrations of 50 - 400 microg/ml for 48 h reduced to 49.6% +/- 3.5%, 30.3% +/- 3.8%, 17.7% +/- 2.2%, and 13.0% +/- 2.5% respectively, all significantly lower than that of the control group [(100.0% +/- 0.8%), all P < 0.05]. After treatment with 100 microg/ml EGCG for 24 h, 48 h, 72 h, and 96 h, the live HepG2 cell numbers were (8.0 +/- 1.5), (22.0 +/- 3.1), (37.0 +/- 5.4), and (61.0 +/- 8.7) 10(4) respectively, all significantly lower than those of the control cells [(15.0 +/- 2.5), (45.0 +/- 5.3), (86.0 +/- 11.0), and (210.0 +/- 23.0) 10(4) respectively, all P < 0.05]; and the live SMMC-7721 cell numbers were (7.0 +/- 2.2), (13.0 +/- 2.5), (20.0 +/- 3.7), and (31.0 +/- 4.0) 10(4) respectively, all significantly lower than those of the control cells [(15.0 +/- 2.5), (45.0 +/- 5.3), (86.0 +/- 11.0), and (210.0 +/- 23.0) 10(4) respectively, all P < 0.05]. The apoptotic rates of HepG2 cells treated with EGCG of the concentrations of 50, 100, and 200 microg/ml for 12 h were 8.7% +/- 0.4%, 18.1% +/- 1.1%, and 22.1% +/- 1.8% respectively, all significantly higher than that of the control group (3.3% +/- 0.3%, P < 0.05); and the apoptotic rates of SMMC-7721 cells were 5.9% +/- 0.3%, 7.8% +/- 0.6%, and 12.2% +/- 0.8% respectively, all significantly higher than that of the control group (3.7% +/- 0.4%, P < 0.05). After treatment with EGCG of the concentrations of 100 and 200 microg/ml for 12 h, the caspase-9 activities of the HepG2 cells increased to (1.8 +/- 0.4) and (2.5 +/- 0.4) respectively, both significantly higher than that of the control group (1.0 +/- 0.1, both P < 0.05); and the caspase-3 activities of the HepG2 cells increased to (2.0 +/- 0.4) and (2.8 +/- 0.5) respectively, both significantly higher than that of the control group (1.0 +/- 0.2, P < 0.05) ; and the caspase-9 activities of the SMMC-7721 cells increased to (1.7 +/- 0.4) and (2.5 +/- 0.4), both significantly higher than that of the control group (1.0 +/- 0.1, both P < 0.05), and the caspase-3 activities of the SMMC-7721 cells increased to (1.9 +/- 0.4) and (2.6 +/- 0.3) respectively, both significantly higher than that of the control group [ (1.0 +/- 0.2), both P < 0.05]. When the concentration of EGCG was over 200microg/ml, it down-regulated the expression of COX-2 and Bcl-2 in both cell lines, however, EGCG resulted in no significant changes of Bcl-xl, Bax, Bad, and Bid., Conclusion: EGCG induces apoptosis in HCC cells through down-regulation of COX-2 and Bcl-2 and consequently activating caspase-9 and caspase-3.

Objective: To investigate the mechanism that mda-7/IL-24 selectively kills hepatocellular carcinoma (HCC) HepG2 cells in vitro., Methods: HCC cell line HepG2 and normal liver cell line L02 were infected with Ad.mda-7. The expression of mda-7/IL-24 was detected by RT-PCR and ELISA, respectively. The apoptotic effects were confirmed by Hoechst staining and flow cytometry assay, respectively. Furthermore, Bcl-2 family proteins, cytochrome C, Smac/DIABLO and caspase-9 were determined by Western blot., Results: The exogenous mda-7/IL-24 gene was expressed in HepG2 and L02 cells infected with Ad.mda-7. Ad.mda-7 induced apoptosis in HepG2 but not in L02 cells in vitro. The induction of tumor cell apoptosis is correlated with the increasing expression of Bax and decreasing expression of Bcl-2 and Bcl-xL genes, then facilitated the releasing of cytochrome C and Smac/DIABLO from mitochondria to cytoplasm and increasing the expression of caspase-9, eventually, resulted in apoptosis., Conclusion: Ad.mda-7 selectively induces growth inhibition and apoptosis in hepatocellular carcinoma HepG2 cells but not in normal L02 hepatocytes in vitro, and the mechanism might involve the decrease of Bcl-2 and Bcl-xL and increase of Bak expression, facilitating the release of cytochrome C and Smac/DIABLO from mitochondria in HCC cells.

Objective: To investigate the anticancer effects of betulinic acid (BA) on Jurkat cells in vitro and its molecular mechanism., Methods: The effects of betulinic acid on the growth of Jurkat cells were studied by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5diphenyl-2H-tetrazolium (MTT) assay. Apoptosis was assessed by Hoechst33258 staining and annexin-V/PI double-labeled cytometry. The effect of betulinic acid on the cell cycle of Jurkat cells was studied by propidium iodide staining. RT-PCR and Western blot were used to analyze the changes of cyclin D3, bcl-xl mRNA and protein levels in Jurkat cells after treatment with betulinic acid., Results: The proliferation of Jurkat cells was decreased in betulinic acid-treated group at a 24 h IC50 value of 70.0 micromol/L. The effect of betulinic acid to induce apoptosis in Jurkat cells was in a time- and dose-dependent manner. Jurkat cells treated with betulinic acid showed an increase of G0/G1 phase and decrease of S phase. The Jurkat cells treated with 0, 20, 60, 100 micromol/L betulinic acid for 24 h, showed an increased G0/G1 phase from 31.0% to 58.8%, whereas decreased S phase from 61.5% to 35.8%, respectively. PBMC was less sensitive to the cytotoxic effect of betulinic acid than Jurkat cells. The expression of cyclin D3, bcl-xl mRNA and protein were decreased sharply in Jurkat cells treated with betulinic acid., Conclusion: Betulinic acid can inhibit the proliferation of Jurkat cells by regulating the cell cycle that arrests cells at G0/G1 phase and induces apoptosis in Jurkat cells. The antitumor effects of betulinic acid may be related to down-regulation of the expression of cyclin D3 and bcl-xl.

Objective: To explore the mechanism of Kechuanluo oral liquid, a compound Chinese herbal medicine, in inhibiting allergic airway inflammation by observing the effects of Kechuanluo on eosinophil (EOS) apoptosis and its regulation factors in asthmatic mice., Methods: Fifty-six BALB/c mice were randomly divided into normal control group (n=16), untreated group (n=16), Western medicine group (n=12) and Kechuanluo group (n=12). Except for the mice in normal control group, asthma was induced in BALB/c mice by using ovalbumin (OVA) and potassium aluminium sulfate. The mice were intragastrically administered with normal saline, Kechuanluo (30 ml/kg daily) and prednisolone tablets (10 mg/kg daily) respectively for two weeks. At 0 hour, the 3rd, 7th and 14th day after the end of OVA sensitization, the EOSs of lung tissues were counted by improved-Giemsa staining method; immunohistochemical method and image analysis were used to detect the expressions of Fas, FasL and Bcl-XL in the EOSs in the four groups; and apoptotic rates of the EOSs in the lung tissues of asthmatic mice were detected by terminal deoxynucleotidyl transferase-mediated biotin-dUTP nick end labeling (TUNEL) technique., Results: Compared with the untreated group, airway inflammations of the mice in the Kechualuo group and Western medicine group were lessened and the EOS counts decreased on the 3rd, 7th and 14th day after the last OVA sensitization (P<0.05). On the 3rd day, the EOSs apoptotic rates, the expression areas of Fas in the EOSs and FasL in the lung tissues were significantly higher in the Kechuanluo group than those in the untreated group (P<0.01), furthermore, the EOS apoptotic rate reached the peak level. Inversely, the EOS count and the expression area of Bcl-XL in the EOSs were obviously lower in the Kechuanluo group (P<0.05). On the 7th and 14th day, the expression areas of Fas and Bcl-XL in the EOSs were significantly decreased in the Kechuanluo group (P<0.01), and on the 14th day, the EOS apoptotic rate and the expression area of FasL in the lung tissues were obviously lower either. The effects exhibited in the Western medicine group were similar to those in the Kechuanluo group., Conclusion: There is airway inflammation with eosinophilic infiltration in asthmatic mice, accompanied with suppressed apoptosis and delayed apoptosis. Kechuanluo can induce and accelerate EOS apoptosis in early inflammation by inhibiting eosinophilic inflammation, improving Fas expression in EOSs and FasL expression in the lung tissues, and reducing Bcl-XL expression in EOSs.

Objective: To investigate the reversing effect of Bcl-XL small interfering RNA (siRNA) on the acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in human colon cancer., Methods: Human colon cancer cells DLD1-TRAIL/R, with acquired resistance to TRAIL, were firstly transfected with Bcl-XL siRNA for 24 h followed by the treatment of TRAIL protein. The survival rate of DLD1-TRAIL/R cells was assessed by FACS analysis and cell number counting, respectively, and activation of its apoptotic signaling was evaluated by Western blot., Results: Bcl-XL siRNA effectively downregulated the expression of Bcl-XL protein and reversed the acquired resistance to TRAIL in DLD1-TRAIL/R cells. After combination treatment of Bcl-XL siRNA and TRAIL protein, the apoptotic rate of DLD1-TRAIL/R cells was more than 50% and survival rate was less than 40%, whereas there was no effect on the survival of DLD1-TRAIL/R cells after treatment with control treatment or TRAIL protein treatment alone (P < 0.05). Western blot analysis demonstrated that caspase-8, caspase-9, Bid, caspase-3, and poly (ADP-ribose) polymerase (PARP) were obviously activated after combination treatment with Bcl-XL siRNA and TRAIL protein, and the release of cytochrome C was also significantly increased., Conclusion: Bcl-XL siRNA can effectively reverse the acquired resistance to TRAIL in human colon cancer cells, suggesting that it might be a new strategy for overcoming the resistance in cancer therapy.

This study was aimed to investigate the effect of STI571, an inhibitor of tyrosine kinase, on the proliferation and apoptosis of chronic myelogenous leukemia (CML) cells as well as expression of anti-apoptotic protein Mcl-1, and to explore the possible role of Mcl-1 in apoptosis-inducing mechanism. K562 cell line was used to observe the effect of STI571 on CML cells. Proliferation and cytotoxicity were analyzed by MTT assay. The apoptotic cells were labelled with Annexin V-FITC and PI and then analyzed by flow cytometry. The expression of apoptotic-related proteins in K562 cells was determined by Western blot with specific antibodies. The results showed that STI571 significantly inhibited the proliferation and induced apoptosis of K562 cells in a dose-and time-dependent manner. Coincidently, the protein phosphorylation on tyrosine residues was reduced and the expressions of anti-apoptotic protein Mcl-1 and Bcl-xl were down-regulated after exposure to STI571. It is concluded that STI571 induces the apoptosis of CML cells by down-regulating the expressions of Mcl-1 and Bcl-xl, which suggests that Mcl-1 and Bcl-xl may play an important role in anti-apoptotic process of CML cells.

Objective: To investigate the expression status and clinical significance of cytoprotective proteins to human renal allografts., Methods: Expressions of cytoprotective proteins: antiapoptotic protein (A20), hemeoxygenase-1 (HO-1), and B-cell lymphoma/leukemia-X(L) (Bcl-X(L)) were analyzed by immunohistochemistry and compared in 30 renal allografts, including 10 grafts undergoing acute rejection (AR), 10 grafts undergoing chronic rejection (CR), and 10 nonrejecting (NR) grafts., Results: Expressions of A20, HO-1 and Bcl-X(L) localized mainly in endothelial, smooth muscle, and infiltrating mononuclear cells. A20 expressed in grafts undergoing AR (5.04 +/- 0.71) and CR (3.20 +/- 0.64), not in NR grafts, and its expression in CR was weaker than that in AR ( P < 0.01). HO-1 expressed in AR (7.91 +/- 2.24), not in CR and NR. Bcl-X(L) was detected in all grafts (AR: 10.62 +/- 3.17; CR: 8.50 +/- 2.45; NR: 11.03 +/- 2.77), but had a decreased expression levels in CR., Conclusion: A20 and HO-1 may play protective role for AR, and A20 may be the essential protein having protective function for CR.

Objective: To explore the protective effect of neuregulin against cardiac myocyte injury in pacing-induced heart failure in rhesus monkeys and its mechanism., Methods: Rapid pacing was used to induce heart failure in rhesus monkeys. Aorta intubation was used to perform hemodynamic measurements such as the peak positive rate of of left ventricular pressure (LVdP/dtmax) and left ventricular systolic, end-diastolic blood pressures (LVSP and LVEDP, respectively) 17 days after the pacing. N-terminal pro-brain natriuretic peptide (BNP), as molecular marker of heart failure, was also measured by electrochemical luminescence immunoassay. The apoptosis of cardiac myocyte was observed by Tunel method. Western blot was used to detect the PKB activity and the relative amounts of Bcl-xl protein in the left ventricular free walls., Results: After the daily intravenous injection of 3microg/kg recombinated neuregulin for 10 days, LVdP/dtmax increased significantly while BNP decreased remarkably. The apoptotic index was obviously lower, and Bcl-xl and activity of PKB were higher., Conclusion: Neuregulin protects against rapid pacing-induced apoptosis in heart failure in rhesus monkeys and the mechanism might be attributed to the increase of the activity of PKB and Bcl-xl protein.

Objective: To investigate the effect of complete Freund's adjuvant (CFA) on islet beta cell apoptosis in preventing diabetes in non-obese diabetic (NOD) mice, and the influence on apoptotic related-gene expression., Methods: Four-week-old female NOD mice were randomly divided into Group CFA (n=5) and Group saline (NS) control (n=5). Mice in Group CFA were injected in the hind footpad with 50 microL CFA and mice in Group NS with 50 microL NS. Blood sugar was monitored and diabetes was diagnosed if blood sugar was higher than 11.1 mmol/L for 2 continuous days in the NOD mice. The mice were sacrificed when diagnosed as diabetes or at 30 weeks of age. Pancreatic sections were made for: evaluation of insulitis severity with HE staining; counting of apoptotic beta cells with the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) method, and ABC immunohistochemical double labeling; counting of Fas, FasL and Bcl-x positive cells respectively with ABC immunohistochemical method., Results: By 30 weeks of age, none of the 5 CFA-treated mice, compared with 3 of the 5 control mice, had developed diabetes. The insulitis score was lower (1.820+/-0.962 vs. 3.020+/-1.040, P<0.05), the rates of apoptotic beta cells, Fas positive cells and FasL positive cells were lower [(10.2+/-2.8)% vs. (15.9+/-6.5)%, (54.9+/-14.5)% vs. (75.7+/-12.9)%, (20.3+/-10.4)% vs. (27.9+/-12.0)%, P<0.05), and the Bcl-x positive cell rate was higher [(74.9+/-10.7)% vs. (66.0+/-18.3)%, P<0.05] in the CFA-treated group than those in the NS-treated group respectively., Conclusion: CFA may inhibit beta cell apoptosis in NOD mice by regulating Fas, FasL and Bcl-x expression on cells within islets.

Objective: To investigate the mechanism of myocardial cell apoptosis during delayed preconditioning induced by Sini decoction (SND)., Method: SD rats were divide into four groups: control, sham, I/R and SND groups. The rats in I/R group, left anterior descending coronary artery (LAD) was occluded for 1h and reperfused for 1 h. The rats in SND group were pretreated with Sini decoction (5 g x kg(-1) x d(-1)) for three days, the last treatment was pretreated 24 h before the index occlusion. Cell apoptosis was measured by flow cytometry, cytochrome C and bcl-xl were detected by Western blotting and the activity of caspase-3 was detected by assay kit., Result: As compared with I/R group, apoptosis rate of myocardial cell, the release of cytochrome C from mitochondria and the activity of caspase-3 were significantly decreased, and the expression of bcl-xl protein was elevated in SND group., Conclusion: The delayed preconditioning induced by Sini decoction decreased myocardial cell apoptosis. The mechanism may be related to the inhibition of mitochondria signal pathway of apoptosis.

Objective: To overcome the drug-resistance of tumor cells is one of the methods of improving the therapeutic results. Histone deacetylase inhibitors (HDACIs) is a novel class of chemotherapeutic agents which can induce apoptosis of tumor cells. Valproic acid (VPA) is a common drug used in the treatment of epilepsy. It has been shown that VPA has a marked HDACIs effect at the pharmaceutical level, and can induce the differentiation and apoptosis of transformed cells. But the mechanism of its effect has not been clarified. The aim of this study was to investigate the mechanism of VPA in inducing the apoptosis of leukemic cells at molecular level., Methods: The cell lines U937, Jurkat clone E6 - 1 (Jurkat) and BALL-1 were cultured in RPMI 1640 medium containing 20% calf serum, then divided into three groups (control group, 1 mmol/L VPA group and 1 mmol/L VPA + 1 micromol/L Pan-caspase inhibitor zVAD-fmk group). At 72 hours after the treatment, the cells were double stained with Aunexin and PI (propidium iodide) and then were analyzed with the flow cytometry (FCM) to detect apoptosis. Before and after treatment with VPA the mean fluorescence index (MFI) of Bcl-2, Bax, Bcl-xl and the levels of caspase 8, 9 and 3 were also detected with the FCM. The changes of P(44/42) mitogen activating protein kinase (MAPK) and phosphorylated P(44/42) MAPK were determined by Western blotting., Results: Seventy-two hours after the treatment, 1 mmol/L VPA induced apoptosis of U937 and Jurkat. The apoptotic rate of U937 was (75.78 +/- 4.20)% and that of Jurkat was (53.50 +/- 5.87)% (P < 0.01, vs. control group); zVAD-fmk could fully inhibit the apoptosis of U937, and the apoptotic rate was (2.89 +/- 0.36)%; while it could partly inhibit the apoptosis of Jurkat, and the apoptotic rate was (15.38 +/- 1.40)% (P < 0.01). 1 mmol/L VPA could not induce the apoptosis of BALL-1 which had a high expression level of Bcl-2. The MFI of Bcl-2, Bax and Bcl-xl in these three cell lines did not change significantly with VPA (P > 0.05). After treatment with VPA, the level of caspase 3 in U937 increased from (14.09 +/- 1.19)% to (32.30 +/- 2.47)%, and caspase 8 from (4.58 +/- 1.41)% to (86.47 +/- 3.26)% (P < 0.01), but there was no significant change in caspase 9 [(13.25 +/- 3.11)% and (10.95 +/- 1.30)%]. In Jurkat, the level of caspase 3 increased from (12.01 +/- 1.63)% to (35.56 +/- 0.27)%, and caspase 9 from (13.89 +/- 1.71)% to (75.89 +/- 4.08)% (P < 0.01 for both); no significant change was observed for caspase 8 [(5.94 +/- 1.38)% and (5.44 +/- 0.72)%]. In BALL-1, there was a slight decline in caspase 3 (P < 0.05). With the effect of VPA, levels of P(44/42) MAPK and phosphorylated P(44/42) MAPK decreased in all three cell lines (P < 0.01)., Conclusion: VPA could induce apoptosis of U937 through the activation of caspase 3 and 8; and it induced the apoptosis of Jurkat involving the activation of caspase 3 and 9. P(44/42) MAPK pathway also plays an important role in this course. VPA induced apoptosis of these cell lines without the alteration of Bcl-2, Bax and Bcl-xl. High level of Bcl-2 could antagonize the effect of VPA.