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Gliomas are the most common central nervous system tumours; they are highly aggressive and have a poor prognosis. RGS16 belongs to the regulator of G-protein signalling (RGS) protein family, which plays an important role in promoting various cancers, such as breast cancer, pancreatic cancer, and colorectal cancer. Moreover, previous studies confirmed that let-7c-5p, a well-known microRNA, can act as a tumour suppressor to regulate the progression of various tumours by inhibiting the expression of its target genes. However, whether RGS16 can promote the progression of glioma and whether it is regulated by miR let-7c-5p are still unknown. Here, we confirmed that RGS16 is upregulated in glioma tissues and that high expression of RGS16 is associated with poor survival. Ectopic deletion of RGS16 significantly suppressed glioma cell proliferation and migration both in vitro and in vivo. Moreover, RGS16 was validated as a direct target gene of miR let-7c-5p. The overexpression of miR let-7c-5p obviously downregulated the expression of RGS16, and knocking down miR let-7c-5p had the opposite effect. Thus, we suggest that the suppression of RGS16 by miR let-7c-5p can promote glioma progression and may serve as a potential prognostic biomarker and therapeutic target in glioma.
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Humans , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , MicroRNAs/metabolism , Glioma/genetics , Genes, Tumor Suppressor , Cell Proliferation , Gene Expression Regulation, Neoplastic , Cell Line, TumorABSTRACT
Objective:To screen important genes and characterize their functions and signaling pathways by weighted gene co-expression network analysis (WGCNA) on the gene expression profile of brain tissue after traumatic brain injury (TBI) so as to provide a reference for the mechanism research and treatment of TBI.Methods:The rat TBI gene expression profile GSE2871 was downloaded from the Gene Expression Omnibus (GEO). The expression profile of 8 799 genes of all 47 rat brain tissue samples was analyzed by WGCNA. After calculating and selecting the β-weighted soft threshold, undirected weighted gene network was constructed to identify gene sets with a high degree of correlation. Sample information was obtained from the database to calculate the correlation between each trait of the samples and modules. Gene ontology (GO) analysis and KEGG pathway analysis were performed for the genes in modules related to injury severity and sampling side in order to unvail the biological processes and pathways involved. The gene-module correlation and gene-trait correlation in these key modules were calculated and hub genes were selected.Results:All the rat brain tissue samples and genes in GSE2871 were included in the WGCNA analysis. A total of 22 modules were obtained, which were marked as modules A to V. Modules E, G, T and U were significantly associated with the sampling side. Modules E and G were significantly related to injury severity . GO analysis and KEGG pathway analysis indicated that the genes in modules E and G with significant relation to injury severity and sampling side were mainly implicated in leukocyte migration, cell chemotaxis, various immune response regulation, etc. The involved pathways included antigen processing and presentation pathways, cell factor-cytokine receptor interaction, interleukin-17 signaling pathway, etc. While modules T and U with significant relation to the sampling side were mainly implicated in hypoxia response, cell metabolism, cell membrane ion channel regulation, signal transduction, etc. The pathways involved were neurodegenerative disease signaling pathways, ribosomes, autophagy, neuroactive ligand-receptor interactions, etc. Among the key modules significantly relating to traits, Tuba1b/1c, Ifitm3, Cebpd, Nfkbia, Serinc3, Pmpcb and Cyp4a8 were selected as hub genes of the above key modules.Conclusion:The genes significantly relating to rat TBI are mainly involved in pathophysiological links such as immune activation, inflammatory response, abnormal energy metabolism, calcium channel disorders, abnormal autophagy and cell apoptosis.
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Objective:To investigate the humoral immune response to GⅠ.1/GⅡ.4 norovirus (NoV) virus-like particle (VLP) vaccine of different doses in mice.Methods:The GⅠ.1/GⅡ.4 norovirus vaccine was diluted into four different concentrations, containing 50, 25, 8.3 and 2.8 μg/dose antigens, respectively. Aluminum hydroxide adjuvant was used in the control group. Ten 7-week-old BALB/c mice in each group were immunized intraperitoneally with 0.5 ml vaccine or adjuvant on 0 d and 21 d. Blood samples were collected on 35 d and the histoblood group antigen (HBGA) blocking titers of GⅠ.1 and GⅡ.4 antibodies in serum were detected. The differences in antibody levels between the groups were analyzed by SPSS23.0 software.Results:GⅠ.1 and GⅡ.4 HBGA blocking antibodies in 50, 25 and 8.3 μg/dose groups became positive on 35 d after the first dose vaccination. The geometric mean titers (GMT) of GⅠ.1 and GⅡ.4 HBGA blocking antibodies were 488 (95%CI: 249-955) and 489 (95%CI: 302-790) in 50 μg/dose group, 278 (95%CI: 106-728) and 738 (95%CI: 299-1 820) in 25 μg/dose group, 300 (95%CI: 158-570) and 486 (95%CI: 222-1 068) in 8.3 μg/dose group, respectively. The positive rates of GⅠ.1 and GⅡ.4 blocking antibodies in 2.8 μg/dose group were 40% and 70% and the GMT were 23 (95%CI: 10-51) and 85 (95%CI: 24-304), respectively. The GⅠ.1 and GⅡ.4 blocking antibodies in the control group were negative. Statistically significant differences in antibody levels were found between 50, 25, 8.3 μg/dose groups and the control group ( P<0.05), as well as in GⅠ.1 blocking antibodies between 50, 25, 8.3 μg/dose groups and 2.8 μg/dose group ( P<0.05). GⅡ.4 antibody level in 25 μg/dose group was statistically different from that in 2.8 μg/dose group ( P<0.05). Conclusions:GⅠ.1/GⅡ.4 norovirus VLP vaccine at (50-8.3) μg/dose could induce humoral immune response in mice.
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Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.
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Objective@#To determine the clinical benefits of internal mammary sentinel lymph node biopsy (IM-SLNB) acquired by breast cancer patients with clinically positive axillary lymph node (ALN), and further optimize the IM-SLNB indications.@*Methods@#All primary breast cancer patients with clinically positive ALN from February 2014 to September 2017 were prospectively recruited in this study. IM-SLNB was performed under the guidance of the modified injection technique. The success rate and visualization rate of IM-SLNB, metastatic rate of internal mammary sentinel lymph node (IMSLN) and its related factors were analyzed, and the clinical benefits were accessed according to the current guidelines.@*Results@#Among 126 patients, all of 94 patients (74.6%) who showed internal mammary drainage successfully underwent IM-SLNB. The incidence of internal mammary artery bleeding and pleural lesion were 4.3%(4/94) and 9.6%(9/94), respectively. The metastatic rate of IMSLN was 38.3% (36/94), which was significantly associated with the number of positive ALN (P<0.001) and tumor size (P=0.024). The lymph node staging of 94 patients who underwent IM-SLNB was more accurate. Among them, 36 cases with positive IMSLN underwent internal mammary radiotherapy (IMRT), while the other 58 cases with negative IMSLN avoided radiotherapy.@*Conclusions@#IM-SLNB should be routinely performed in patients with positive ALN. IM-SLNB can provide more accurate staging and guide tailored IMRT to benefit more breast cancer patients.
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Objective@#To evaluate whether simultaneous vaccination with live attenuated polio vaccine affects the immunogenicity of live attenuated rotavirus (RV) vaccine.@*Methods@#Rotarix produced by GlaxoSmithKline was used as the research object. Two doses of Rotarix were orally administered on day 0 and month 1, and oral live attenuated polio vaccine (OPV) was administered on day 0, month 1 and month 2 according to the national vaccination plan. Healthy infants aged 6 to 16 weeks were randomly divided into two groups: interval vaccination group (Rotarix and OPV were vaccinated on different days) and simultaneous vaccination group (Rotarix and OPV were vaccinated on the same day). Serum samples were collected on day 0, month 2 and month 12, and serum RV-IgA was measured by enzyme linked immunosorbent assay. Statistical analysis was performed to evaluate whether there were statistical differences in the seroconversion rate and level distribution of RV-IgA between the two groups.@*Results@#The seroconversion rate of serum RV-IgA in month 2 was 73.84% in the interval vaccination and 63.95% in the simultaneous vaccination group, and the difference between them was statistically significant (P<0.05). The geometric mean concentrations (GMC) of RV-IgA were 97 EU/ml and 90 EU/ml, respectively (P>0.05). Compared with the simultaneous vaccination group, the seroconversion rate and GMC of serum RV-IgA in month 12 were higher in the interval vaccination group, and the differences were statistically significant (P<0.05).@*Conclusions@#Simultaneous vaccination with live attenuated polio vaccine would affect the immune response of live attenuated rotavirus vaccine, especially the maintenance of RV-IgA antibody level.
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Objective To grasp the current situation of drinking water-borne endemic fluorosis in Shandong Province,and provide a scientific basis for formulating preventive strategies.Methods In 2016,according to the "National Surveillance Scheme of Drinking Water-Borne Endemic Fluorosis" and "Shandong Provincial Surveillance Scheme of Drinking Water-Borne Endemic Fluorosis",10 counties (cities,districts) were selected in Shandong Province,3-5 villages were selected in each county (city,district),to investigate the operation of water improvement and fluorine reduction project and the water fluorine,and 1 tap water sample in each village was collected to detect water fluorine.At the same time,dental fluorosis examination was performed on all children aged 8-12 years old and fluorosis X-ray examination was performed on adults aged ≥25 years old.Water fluorine was detected by ion selective electrode method;the water fluorine was determined according to "Standards for Drinking Water Quality" (GB 5749-2006);criteria for diagnosis,grading and epidemic intensity of dental fluorosis were based on "Diagnosis of Dental Fluorosis" (WS/T 208-2011);the diagnostic and grading criteria for skeletal fluorosis were based on the "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS 192-2008).Results All the 32 villages in 10 counties (cities,districts) changed water,water improvement and fluorine reduction project ran normally,24 villages had qualified water fluorine (≤ 1.20 mg/L),and 8 villages had water fluorine exceeding the standard.The detection rate of dental fluorosis in children was 25.75％ (419/1 627),and the difference between 10 counties (cities,districts) was statistically significant (x2 =288.55,P ＜ 0.01);the dental fluorosis index was 0.48,and the defect rate was 2.77％ (45/1 627).The detection rate of skeletal fluorosis in adults was 4.72％ (72/1 525),and the difference between 10 counties (cities,districts) was statistically significant (x2 =18.83,P ＜ 0.01).Conclusions The problem of water fluorine exceeding the standard in water improvement and fluorine reduction project in Shandong Province is still serious.The condition of fluorosis has not been fully controlled,and the prevention and treatment measures need to be further strengthened.
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Objective To evaluate whether simultaneous vaccination with live attenuated polio vaccine affects the immunogenicity of live attenuated rotavirus ( RV) vaccine. Methods Rotarix produced by GlaxoSmithKline was used as the research object. Two doses of Rotarix were orally administered on day 0 and month 1, and oral live attenuated polio vaccine (OPV) was administered on day 0, month 1 and month 2 according to the national vaccination plan. Healthy infants aged 6 to 16 weeks were randomly divided into two groups:interval vaccination group ( Rotarix and OPV were vaccinated on different days) and simultane-ous vaccination group ( Rotarix and OPV were vaccinated on the same day) . Serum samples were collected on day 0, month 2 and month 12, and serum RV-IgA was measured by enzyme linked immunosorbent assay. Statistical analysis was performed to evaluate whether there were statistical differences in the seroconversion rate and level distribution of RV-IgA between the two groups. Results The seroconversion rate of serum RV-IgA in month 2 was 73. 84％ in the interval vaccination and 63. 95％ in the simultaneous vaccination group, and the difference between them was statistically significant (P<0. 05). The geometric mean concen-trations (GMC) of RV-IgA were 97 EU/ml and 90 EU/ml, respectively (P>0. 05). Compared with the simultaneous vaccination group, the seroconversion rate and GMC of serum RV-IgA in month 12 were higher in the interval vaccination group, and the differences were statistically significant (P<0. 05). Conclusions Simultaneous vaccination with live attenuated polio vaccine would affect the immune response of live attenua-ted rotavirus vaccine, especially the maintenance of RV-IgA antibody level.
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Objective:To explore the relationship among emotional intelligence,emotional labor and job satisfaction.Methods:Totally 257 bus drivers [158 males and 99 females,aged 17 to 50 years,mean age (32 ± 7)years] in Jinan were surveyed with the Emotional Intelligence Scale (EIS),the Emotional Labor Scale (ELS) and the Minnesota Satisfaction Questionnaire (MSQ).Bootstrap was used to test the mediating effect.Results:The scores of EIS and ELS were both positively correlated with the MSQ scores (r =0.28,0.23,Ps ＜0.01) in the bus drivers.The surface acting (SA) scores were negatively correlated with the MSQ scores (r =-0.18,P ＜ 0.01),and the deep acting (DA) score were positively correlated with the MSQ scores (r =0.40,P ＜ 0.01).Mediating effect test showed that the indirect effect of surface acting was 0.07 (95％ CI:0.01-0.16),and the indirect effect of deep acting was 0.10 (95％ CI:0.03-0.21).Correspondingly,the direct effect of emotional intelligence on job satisfaction respectively was 0.35 (95％ CI:0.20-0.58) and 0.33 (95％ CI:0.12-0.53).Conclusion:These findings support that the bus drivers with higher emotional intelligence may be more satisfied with their job.Emotional intelligence is found to indirectly influence job satisfaction through surface acting and deep acting,and it may affect the bus drivers more effectively choose and use emotion regulation strategies.
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Objective To investigate the clinical efficacy and safety of re-radiotherapy combined with TS-1 (tegafur gimeracil oteracil potassium capsule) in treatment of local recurrent esophageal carcinoma.Methods A total of 63 esophageal carcinoma patients who recurred after the first course radiation treatment admitted to Jiangdu People's Hospital of Yangzhou during January 1,2012 to June 30,2015 were retrospectively analyzed.Twenty-seven of them treated with re-radiotherapy combined with TS-1 were deemed as the research group and 36 of them treated with radiotherapy alone were deemed as the control group based on different treatment.Then the clinical efficacy and adverse reactions of the two groups were compared.Results The objective response rates were 77.8％ (21/27) and 50.0％ (18/36) respectively in the research group and control group,and the difference was statistically significant (x2 =5.048,P =0.025).The median survival time in the two groups were 21.6 months and 13.7 months,the 1-year (74.1％) and 2-year (44.4％) survival rates of the research group were both higher than those of the control group (52.8％ and 30.6％,respectively),and the difference was statistically significant (x2 =6.086,P =0.013).The major adverse effects of the research group and control group during the treatment were radiation oesophagitis (92.6％ vs.80.5％),radiation pneumonia (18.5％ vs.19.4％),myelosuppression (96.3％ vs.77.8％) and gastrointestinal reactions (25.9％ vs.19.4％).Most of them were 1-2 grade,and there were no statistically significant differences (x2 =0.975,P =0.323;x2 =0.009,P =0.926;x2 =2.941,P =0.086;x2 =0.375,P =0.540).Conclusion The treatment of re-radiotherapy combined with TS-1 for local recurrent esophageal carcinoma can improve the efficacy and prolong survival period,and the adverse reactions are tolerable.
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Objective To understand the implementation effects of control measures and the dynamic prevalence of drinking water-borne endemic fluorosis in Shandong Province.Methods From 2013 to 2017 in Shandong Province drinking water-borne endemic fluorosis areas,in accordance with the method of simple random sampling,10 monitored counties were selected,10 water fluorine decreasing projects were investigated of its running effect,and water fluoride was detected.In each monitored county,3 villages in the diseased areas were selected as fixed monitored villages.If the monitored village had changed its water,the operation of the project would be investigated.If the monitored village had not changed the water,one water sample would be collected according to 5 directions of east,west,south,north and middle to determine the water fluorine.All children aged from 8 to 12 in the village were also examined for dental fluorosis.X-ray fluorosis and urinary fluorosis in adults aged over 25 were detected in 2013 and 2017.Results From 2013 to 2017,in monitored counties,the normal operation rate of improved-water defluoridation projects was 99.19％ (368/371),the qualified rate of water fluoride content was 66.85％ (248/371).In 128 improved water projects,the normal operation rate was 100.00％ (128/128),the qualified rate of water fluoride content was 67.19％ (86/128).The average detection rate of dental fluorosis and community fluorosis index (CFI) in children aged 8-12 were 33.92％ (2 608/7 689) and 0.68 in all monitored counties from 2013 to 2017.The detection rate presented a declining trend year by year (x2 =152.47 P ＜ 0.01).The average detection rate and CFI in the villages with normal operation projects and qualified fluoride content were 26.10％ (1 311/5 023) and 0.47.The detection rate presented a declining trend year by year (x2 =207.67 P ＜ 0.01).The detection rate and CFI in the villages with water improvement projects under abnormal operation or excessive fluoride were 48.65％ (1 297/2 666) and 0.90.The detection rate of dental fluorosis in the villages with normal operation projects and qualified fluoride content was lower than the rate of the villages with water improvement projects under abnormal operation or excessive fluoride (x2 =193.98,P ＜ 0.01).In 2013 and 2017,X-ray detection rates of skeletal fluorosis in adults aged over 25 were 8.04％ (119/1 481) and 6.04％ (81/1 342),respectively.The adult urinary fluoride geometric mean values (1.24,1.49 mg/L) were lower than the normal limit of 1.6 mg/L both in qualified villages of water ffuoride content and exceeding-standard villages in 2017.Conclusions In Shandong Province,the effective water improvement has acquired some effects in control of drinking water-borne endemic fluorosis.But the water fluoride content of improved water defluoridation projects exceeds the national standard seriously and fluorosis has not been effectively controlled.
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Objective To evaluate the effect and the anti-tumor mechanism of nucleoside combination on human hepatoma cell line Bel-7402 with Arg-Gly-Asp sequence labeled by ultrasmall superparamagnetic iron oxide (RGD-USPIO).Methods The tumor cells Bel-7402 of logarithmic phlyhase were divided into experimental group and control group,treated with 1 mmol/L nucleoside combination and 1640 medium respectively.The two group were co-cultured for 48 h,and were added RGD-USPIO and co-cultured for 6 h.Then the two groups were proceeded with MR scanning,and the signal intensity of T2WI were measured.After extraction of the total RNA and protein of experiment group and control group,the expression of integrin avβ3 was detected using real-time PCR and Western blot.Results The T2WI signal intensity of experimental group (997.35±42.83) was higher than that of control group (241.05±15.36,t 28.79,P＜0.01).Compared with control group,the expression of integrin αvβ3 mRNA in experimental group was (0.22±0.02) times (t=4.50,P＜0.01).According to Western blot,the protein bands of experimental group were relatively lighter than that of control group,the expression of integrin αvβ3 in experimental group was lower (t =11.88,P＜0.01).Conclusion Nucleoside combination has anti-tumor effect by inhibiting integrin ligand-receptor binding,and the anti tumor mechanism may be related to the induction of tumor cell apoptosis.MR molecular probes can conveniently and accurately evaluate the anti-tumor effect of nucleoside combination on Bel-7402 cells.
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<p><b>OBJECTIVE</b>The aim of this study was to determine the impact of routinely performed internal mammary sentinel lymph node biopsy (IM-SLNB) on the staging and treatment, and to analyze the success rate, complications and learning curve.</p><p><b>METHODS</b>All patients with biopsy-proven breast cancer who underwent sentinel lymph node biopsy between 2012 and 2014 were included in a prospective analysis. Internal mammary sentinel lymph node biopsy (IM-SLNB) was performed in all patients with IM-SLN visualized on preoperative lymphoscintigraphy and/or detected by intraoperative gamma probe detection. The adjuvant treatment plan was adjusted according to the current guidelines.</p><p><b>RESULTS</b>In a total of 349 patients, 249 patients (71.1%) showed internal mammary drainage. IM-SLNB was performed in 153 patients with internal mammary drainage, with a success rate of IM-SLNB of 97.4% (149/153). Pleural lesion and internal mammary artery bleeding were found in 7.2% and 5.2% patients, respectively. In 8.1% of patients (12/149) the IM-SLN was tumor positive. In the group of patients who underwent IM-SLNB, lymph node staging was changed in 8.1% of patients, and IMLNs radiotherapy was guided by these results, however, systemic treatment was changed in only 0.7% of the patients.</p><p><b>CONCLUSIONS</b>IM-SLNB has a high successful rate and good safety. Identification of internal mammary metastases through IM-SLNB may provide more accurate staging and guide the tailored internal mammary radiotherapy.</p><p><b>TRIAL REGISTRATION</b>ClinicalTrials. gov, NCT01642511.</p>
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Female , Humans , Breast , Pathology , Breast Neoplasms , Pathology , Learning Curve , Lymph Nodes , Pathology , Lymphatic Metastasis , Mammary Arteries , Neoplasm Staging , Methods , Neoplasms, Second Primary , Prospective Studies , Sentinel Lymph Node Biopsy , MethodsABSTRACT
<p><b>OBJECTIVE</b>To explore the efficacy and safety of pemetrexed in the treatment of relapsed primary central nervous system lymphoma (PCNSL).</p><p><b>METHODS</b>Seven cases with relapsed PCNSL admitted in our hospital between August 2012 and August 2013 were retrospectively reviewed.</p><p><b>RESULTS</b>Of the 7 relapsed cases, ectopic recurrence occurred in 3, in situ recurrence in 3 and leptomeningeal metastasis in 1. Patients with relapsed PCNSL were administered with high-dose pemetrexed (900 mg/m²) once for every 3 weeks and supplemented with folic acid and vitamin B₁₂. Complete remission was obtained in 2 patients, partial remission in 3 patients and progressive disease in 2. The overall response rate was 71.4% (5/7). The main adverse reactions were myelosuppression and gastrointestinal reaction.</p><p><b>CONCLUSION</b>Treatment of relapsed PCNSL is difficult, and its prognosis is very poor. Pemetrexed therapy is a meaningful trial.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Central Nervous System Neoplasms , Drug Therapy , Pathology , Glutamates , Therapeutic Uses , Guanine , Therapeutic Uses , Lymphoma, Large B-Cell, Diffuse , Drug Therapy , Pathology , Neoplasm Recurrence, Local , Pemetrexed , Prognosis , Retrospective StudiesABSTRACT
Objective:To investigate the influence of Mannan Peptide combined with 5-Fu on the proliferation and apoptosis of HepG2 cells and to explore the related mechanism.Methods:HepG2 cells were divided into four groups:the control group,5-Fu group,Mannan Peptide group and combination group.Cell proliferation was evaluated by MTT assay.Flow cytometry was used to examine the cell cycle,proliferation index(PI)and apoptosis ratio.The expression of Bcl-2 protein in each group was analyzed by immunocytochemical method.Results:The inhibitive effect of 5-Fu plus Mannan Peptide was significantly stronger than that of Mannan Peptide alone,in a time-and dose-dependent manner. Cells were treated with medicine for 48 hours.Compared with cells in the control group and 5-Fu group,the percentage of cells in G_0/G_1 stage and the apoptosis ratio in the combination group were increased(P<0.01).In immunocytochemical analysis,the combination of Mannan Peptide and 5-Fu could significantly decrease Bcl-2 protein level(P<0.01).Conclusion:The combination of Mannan Peptide and 5-Fu can synergistically suppress the proliferation and induce apoptosis of human hepatoma HepG2 cells.The mechanism might be associated with down-regulation of Bcl-2 protein expression.