Pentoxifylline protects against cerebral ischaemia-reperfusion injury through ferroptosis regulation via the Nrf2/SLC7A11/GPX4 signalling pathway.

OBJECTIVE: To investigate whether pentoxifylline (PTX) attenuates cerebral ischaemia-reperfusion injury (IRI) in rats by inhibiting ferroptosis and to explore the underlying molecular mechanisms. METHODS: Cerebral IRI was induced in male Sprague-Dawley (SD) rats using middle cerebral artery occlusion (MCAO). The effects of PTX on cerebral ischaemia-reperfusion brain samples were detected through neurological deficit score, staining and electron microscopy; levels of ferroptosis biomarkers from brain samples were detected using kits. Additionally, the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), transferrin receptor protein 1, divalent metal transporter 1, solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) were determined by immunohistochemistry, real-time quantitative polymerase chain reaction and western blotting. RESULTS: Pre-treatment with PTX was found to improve neurological function, evidenced by reduced neurological deficit scores, decreased infarct volume and alleviated pathological features post-MCAO. This improvement was accompanied by reduced lipid peroxidation levels and mitigated mitochondrial damage. Notably, PTX's inhibitory effect on ferroptosis was characterised by enhanced Nrf2 nuclear translocation and regulation of ferroptosis-related proteins. Moreover, inhibition of Nrf2 using ML385 (an Nrf2-specific inhibitor) reversed PTX's neuroprotective effect on MCAO-induced ferroptosis via the SLC7A11/GPX4 signalling pathway. CONCLUSIONS: Ferroptosis is evident following cerebral ischaemia-reperfusion in rats. Pentoxifylline confers protection against IRI in rats by inhibiting ferroptosis through the Nrf2/SLC7A11/GPX4 signalling pathway.

Effect of La2 O3 and CeO2 on anaerobic methanogenesis process: Kinetics analysis and process mechanism study.

La2 O3 and CeO2 , as main rare earth oxides, with unique physical and chemical properties have been widely used in catalyst and grinding industry. In this study, the effects of La2 O3 and CeO2 on the anaerobic process were investigated. The biological methane production tests showed that 0-0.05 g/L La2 O3 and 0-0.05 g/L CeO2 enhanced anaerobic methanogenesis process. The result showed maximum specific methanogenic rates of La2 O3 and CeO2 were 56.26 mL/(h·gVSS) and 49.43 mL/(h·gVSS) and, compared with the control, increased 4% and 3%, respectively. La2 O3 significantly reduced the accumulation of volatile fatty acids (VFAs), whereas CeO2 had no similar effect. Dissolution experiments demonstrated that the content of extracellular La in the anaerobic granular sludge reached 404 µg-La/g volatile suspended solid (VSS), which was 134 times higher than that of extracellular Ce (3 µg-Ce/gVSS). The content of intracellular La reached 206 µg-La/gVSS, which was 19 times higher than that of intracellular Ce (11 µg-Ce/gVSS). The different stimulation between La3+ and Ce3+ could be attributed to the different dissolution of La2 O3 and CeO2 . The result of this work is helpful to optimize anaerobic processes and to develop novel additives. PRACTITIONER POINTS: Novel anaerobic additives were developed. La2O3 and CeO2 in 0-0.05 g/L enhanced organics degradation and methane production. The addition of La2O3 significantly reduced the accumulation of volatile fatty acids. The solubilization of La2O3 was stronger than CeO2. The promoting effects of low concentrations of La2O3 and CeO2 were derived from dissolved La and Ce.

The novel HLA-DQB1*03:03:29 allele, identified by sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-DQB1*03:03:29 differs from HLA-DQB1*03:03:02:01 by one nucleotide in exon 2.

[Effect of γδ T cells on the Proliferation, Apoptosis and Autophagy of Multiple Myeloma Cells].

AbstractObjective: To investigate the effect of γδ T cells on the proliferation, apoptosis and autophagy of multiple myeloma cells. METHODS: Peripheral blood mononuclear cells (PBMNC) were isolated from healthy volunteers, and stimulated with zoledronic acid (Zol) in combination with rhIL-2. Flow cytometry analysis was used to detected the purity of γδ T cells. γδ T cells were collected and co-cultured with RPMI-8226 or U-266 cells at different effector target ratios. The proliferation of RPMI-8226 or U-266 cell lines were detected by CCK-8. Cell cycle and cell apoptosis were detected by flow cytometry and Western blot．The expressions of autophagy-related proteins were detected by Western blot. RESULTS: γδ T cells can be expanded in vitro. γδ T cells could inhibit the proliferation of RPMI-8226 or U-266 cells, induced cell cycle arrest and promoted apoptosis in an effector target-dependent manner. In addition, γδ T cells could induce autophagy of myeloma cells, inhibited the expression of autophagy-related PI3K, P-AKT and P-mTOR, while increased the expression of AMPK and Beclin-1. CONCLUSION: γδ T cells can inhibit the proliferation of RPMI-8226 and U-266 myeloma cells, induce cell cycle arrest, promote apoptosis, and enhance autophagy in vitro. The mechanism may be related to inhibition of PI3K/AKT/mTOR signaling pathway and/or activation of AMPK/Beclin-1 signaling pathway.

[Genotype Analysis of Pregnant Women with α- and ß- Thalassemia in Fuzhou Area of Fujian Province in China].

OBJECTIVE: To analyze the genotype of pregnant women with α- and ß- thalassemia in Fuzhou area of Fujian province in China. METHODS: Blood routine examination and hemoglobin electrophoresis were performed for pregnant women, and positive samples were examined by gap polymerase chain reaction and reverse dot blot hybridization. RESULTS: 412 cases were diagnosed as α-thalassemia (63.9%); 201 cases were diagnosed as ß-thalassemia (31.2%); 32 cases were diagnosed as α and ß-composite thalassemia. There were 12 genotypes in α-thalassemia, whose major genotypes were --SEA/αα, α3.7/αα, -α4.2/αα and αQSα/αα, with carrying rate of 64.32%, 20.14%, 7.77% and 1.94%, respectively. There were 10 genotypes in ß- thalassemia, whose major genotypes were CD41-42/N, CD17/N, IVS-II-654/N and -28/N, with carrying rate of 30.84%, 27.86%, 15.92% and 10.45%, respectively. There were 9 genotypes in α and ß-composite thalassemia, whose major genotypes were --SEA/αα composited CD41-42/N, -α3.7/αα composited CD41-42/N, --SEA/αα composited CD17/N, with carrying rate of 18.75%, 15.62%, 15.62% respectively. CONCLUSION: The major genotypes of pregnant women with α- and ß- thalassemia in Fuzhou area of Fujian province in China are --SEA/αα, α3.7/αα, CD41-42/N and CD17/N. Thalassemia screening and prenatal gene diagnosis should be strengthened in Fuzhou area of Fujian province in China.

[Research Progress on Regulating Autophagy in the Treatment of Multiple Myeloma --Review].

Abstract　　Autophagy is a process in which cells in eukaryotes degrades abnormal proteins and organelles, thus it possesses important effects on the survival of normal cells and tumor cells. The related studies have shown that autophagy is widely present in the life activities of myeloma cells, which not only protects myeloma cells, but also induces death, and plays an important role in survival, proliferation, invasion and migration of myeloma cells and the treatment of multiple myeloma. This review focuses on the progress of regulating autophagy in the treatment of multiple myeloma.

[Effects of Autophagy Regulating Drugs on Proliferation, Apoptosis and Autophagy of Multiple Myeloma Cells].

OBJECTIVE: To investigate the effects of autophagy activator (rapamycin, RAPA) and autophagy inhibitor (hydroxychloroquine, HCQ and 3-methyl adenine, 3-MA) on the proliferation, apoptosis and autophagy of multiple myeloma cell line of RPMI8226. METHODS: RPMI8226 cells were treated with autophagy regulating drugs of different concentrations. The proliferation and apoptosis of cells were determined by CCK-8 and flow cytometry, respectively. The expressions of apoptosis-related proteins BCL-2, caspase-3 and PARP protein were assessed by Western blot. Autophagy was detected by monodansylcadaverine staining. Autophagic protein (LC-3b) and apoptosis-related proteins (caspase-3, PARP and BCL-2) were analyzed by Western blot. RESULTS: RAPA and HCQ inhibited the proliferation of RPMI8226 in a concentration- and time-dependent manner, and increased the apoptosis. However, 3-MA did not show significantly inhibitory effect on the proliferation and apoptosis of RPMI8226. MDC staining showed that the more autophagic vacuoles could be detected in the higher concentration of RAPA, but the less autophagic vacuoles in the higher concentration of HCQ and 3-MA. Western blot showed that RAPA increased the expression of LC3-II/LC3-I, caspase-3 and PARP, but inhibited the expression of BCL-2. HCQ inhibited the expression of LC3-II/LC3-I and BCL-2, but increased the expression of caspase-3 and PARP. 3-MA inhibited the expression of LC3-II/LC3-I, but had no effect on the expression of caspase-3, PARP or BCL-2. CONCLUSION: Rapamycin can inhibit the proliferation, induce apoptosis and autophagy of RPMI 8226, the hydroxychloroquine can inhibit autophagy and proliferation of RPMI 8226, and induce apoptosis, the 3-MA can inhibit autophagy of RPMI 8226, but hardly has any effects on proliferation and apoptosis of RPMI 8226 cells.

[Significance of H3K27me3 and EZH2 in Predicting the Therapeutic Efficacy of Diffuse Large B-Cell Lymphoma].

OBJECTIVE: To investigate the significance of H3K27me3 and its methyltransferase EZH2 in predicting the short-term and long-term outcome of newly diagnosed patients with diffuse large B-cell lymphoma (DLBCL). METHODS: The paraffin wax speciments of 102 DLBCL patients in Fujian Medical University Cancer Hospital were collected. The expression of H3K27me3, EZH2 and BCL-2 protein were detected using tissue array made by tissue microarray(TMA) technique and immunohistochemistry method. The evaluation data after clinical treatment and follow-up results were collected and combined with expression levels of H3K27me3, EZH2 and BCL-2 detected by tissue array, then on the basis of these data, the survival of patients was analyzed by Kaplan-Meier method, the correlation of EZH2 with H3K27me3 and BCL-2 was analyzed by pearson correlation test, the correlation of above mentioned indicators with different therapeutic efficacy was analyzed by spearman correlation test. The relationship of H3K27me3 and EZH2 expression as well as co-expression of H3K27me3 and EZH2 with the therapeutic efficacy and prognosis of patients were compared. RESULTS: A total of 61.8% patients showed EZH2 high expression which positively correlated with high expression of H3K27me3 and BCL-2. The complete remission (CR) and overall remission (OR) rates in H3K27me3 high expression and co-expression of H3K27me3 EZH2 groups were lower than those in low expression groups (P<0.001), moreover OS and PFS rates also were lower than those in low expression (P<0.001). In the RCHOP subgroup, the patients with EZH2 low expression showed significantly better CR, OR OS and PFS in comparison with those of patients with higher expression (P=0.003,P=0.019). CONCLUSION: Part of DLBCL patients with H3K27me3 high expression or coexpression of both H3K27me3 and EZH2 exhibit a worse prognosis in comparison with those patients with H3K27me3 low expression or without coexpression. The patients with EZH2 low expression usually responde well to RCHOP regimen in the short-term or long-term survival.

Differences in Spontaneous Intracerebral Hemorrhage Cases between Urban and Rural Regions of Taiwan: Big Data Analytics of Government Open Data.

This study evaluated the differences in spontaneous intracerebral hemorrhage (sICH) between rural and urban areas of Taiwan with big data analysis. We used big data analytics and visualization tools to examine government open data, which included the residents' health medical administrative data, economic status, educational status, and relevant information. The study subjects included sICH patients of Taipei region (29,741 cases) and Eastern Taiwan (4565 cases). The incidence of sICH per 100,000 population per year in Eastern Taiwan (71.3 cases) was significantly higher than that of the Taipei region (42.3 cases). The mean coverage area per hospital in Eastern Taiwan (452.4 km²) was significantly larger than the Taipei region (24 km²). The residents educational level in the Taipei region was significantly higher than that in Eastern Taiwan. The mean hospital length of stay in the Taipei region (17.9 days) was significantly greater than that in Eastern Taiwan (16.3 days) (p < 0.001). There were no significant differences in other medical profiles between two areas. Distance and educational barriers were two possible reasons for the higher incidence of sICH in the rural area of Eastern Taiwan. Further studies are necessary in order to understand these phenomena in greater depth.

[Anti-proliferation Effect of NS-398 in Combination with Bortezomib on Multiple Myeloma RPMI 8226 Cells In Vitro].

OBJECTIVE: To investigate if NS-398 could enhance the chemosensitivity of Bortezomib (BOR) on human multiple myeloma RPMI 8226 cells. METHODS: After the treatment of NS-398 combined with BOR, MTT assay was used to detect the proliferation inhibition effect on human multiply myeloma RPMI 8226 cells in vitro, Flow cytometry was used to analyze their effect of apoptosis and cell cycle; the caspase-3 activity of different treatment group was detected by using ELISA and the activity of Cox-2 was measured by using Cox-2 activity assay kit. RESULTS: The inhibitory rate of NS-398 combined with BOR was higher than that of NS-398 or BOR alone(Q>0.85). After treatment of NS-398 combined with BOR, the percentage of cells arrested in the G0/G1 phase and apoptotic rate were both higher than that of treatment with each drug alone(Q>1.15). The caspase-3 activity in cells treated with combined of NS-398 and BOR was significantly higher than that of treatment of each drug alone(Q>1.15). CONCLUSION: NS-398 combined with BOR shows a synergistic effect on the growth inhibition of RPMI 8226 cells in vitro.

A modified molecular framework derived highly efficient Mn-Co-carbon cathode for a flexible Zn-air battery.

A controllable Co doping strategy is introduced to significantly activate more catalytic sites for Mn-based materials and anchor Co-Mn nanoparticles on the N-doped carbon nanotube (N-CNT) substrates. The as-synthesized CoMn2O4/N-CNTs exhibit excellent ORR catalytic performance with large limited current density and positive half-wave potential, even outperforming the Pt/C catalysts. The outstanding ORR activity allows the CoMn2O4/N-CNTs to directly serve as the cathode electrode in a liquid/solid state Zn-air battery, demonstrating large power density and robust stability.

[Effect of Complicated Hemophagocytic Syndrome on Clinical Prognosis of Patients with Non-Hodgkin's Lymphoma and Analysis of Its Affecting Factors].

OBJECTIVE: To investigate the effect of complicatal hemophagocytic syndrome on clinical prognosis of patients with non-Hodgkin's lymphoma (NHL) and analyze its factors affecting prognosis. METHODS: Ninety cases of NHL were selected and divided into 2 groups: 61 cases of NHL without hemophagocytic syndrome as group A and 29 cases of NHL with hemophagocytic syndrame as group B. The survival analysis of Kaplan-Meter method and the Cox regression model were used for univariate and multivariate analyses of related factors. RESULTS: The patients in group B were more likely to start with fever, moreover, the hemophagocytes could be found in bone marrow samples of 89.66% (26/29) patients; the levels of total bilirubin, triglycerides, serum ferritin, serum soluble CD25, DNA copies of epstein-barr virus (EBV) and lactate dehydrogenase level in the group B were significantly higher than those in the group A(P<0.05). And the patients in group B had worse physical state, later disease stage, worse disease status and lower overall prognosis as compared with patients in the group A. The complicased hemophagocytic syndrome, incomplete improvemant of deseases state after treatment and EBV infection were the independent risk factors for the poor prognosis of patients with NHL. CONCLUSION: The complicated hemophagocytic syndrome can increase the severity of NHL, there fore significantly influences the clinical prognosis of patients, while the complicated hemophagocytic syndrome, poor therapatic efficacy for patients and EBV infection are independent risk factors affecting the prognosis of NHL patients.

Clinical significance of expression of proliferating cell nuclear antigen and E-cadherin in gastric carcinoma.

AIM: to investigate the expression of proliferating cell nuclear antigen (PCNA) and E-cadherin in gastric carcinoma and to analyze their clinical significance. METHODS: A total of 146 patients were selected for this study, including 38 patients with intestinal metaplasia, 42 with dysplasia, and 66 with primary gastric cancer. In addition, 40 patients with normal gastric tissues were selected as controls. The expression of PCNA and E-cadherin was detected by immunohistochemistry. Differences in PCNA and the E-cadherin labeling indexes among normal gastric mucosa, intestinal metaplasia, dysplasia, and gastric carcinoma were compared. Subjects with normal gastric tissues were assigned to a normal group, while gastric cancer patients were assigned to a gastric cancer group. The difference in PCNA and E-cadherin expression between these two groups was compared. The relationship between expression of PCNA and E-cadherin and clinicopathological features was also explored in gastric cancer patients. Furthermore, prognosis-related factors, as well as the expression of PCNA and E-cadherin, were analyzed in patients with gastric cancer to determine the 3-year survival of these patients. RESULTS: The difference in PCNA and the E-cadherin labeling indexes among normal gastric mucosa, intestinal metaplasia, dysplasia, and gastric carcinoma was statistically significant (P < 0.05). During the transition of normal gastric mucosa to gastric cancer, the PCNA labeling index gradually increased, while the E-cadherin labeling index gradually decreased (P < 0.05). The PCNA labeling index was significantly higher and the E-cadherin labeling index was significantly lower in gastric cancer than in dysplasia (P < 0.05). The expression of PCNA was significantly higher in the gastric cancer group than in the normal group, but E-cadherin was weaker (P < 0.05). There was a negative correlation between the expression of PCNA and E-cadherin in gastric carcinoma (r = -0.741, P = 0.000). PCNA expression differed significantly between gastric cancer patients with and without lymph node metastasis and between patients at different T stages. E-cadherin expression also differed significantly between gastric cancer patients with and without lymph node metastasis (P < 0.05). High T stage and positive PCNA expression were risk factors for the prognosis of patients with gastric cancer (RR > 1), while the positive expression of E-cadherin was a protective factor (RR < 1). The sensitivity, specificity, and accuracy of PCNA positivity in predicting the 3-year survival of patients with gastric cancer were 93.33%, 38.89%, and 0.64, respectively; while these values for E-cadherin negativity were 80.0%, 41.67%, and 0.59, respectively. When PCNA positivity and E-cadherin negativity were combined, the sensitivity, specificity, and accuracy were 66.67%, 66.67%, and 0.67, respectively. CONCLUSION: Combined detection of PCNA and E-cadherin can improve the accuracy of assessing the prognosis of patients with gastric cancer.

Serum levels of brain-derived neurotrophic factor are associated with depressive symptoms in patients with systemic lupus erythematosus.

OBJECTIVE: The aim of this study was to explore potential relationships between serum BDNF levels and depression in systemic lupus erythematosus (SLE) patients. METHODS: We included 208 consecutive SLE patients and 100 age-and sex-matched healthy controls. The presence of depressive symptoms was determined through the Beck Depression Inventory-II (BDI-II) score. RESULTS: The serum BDNF levels were significantly (P<0.0001) higher in SLE patients as compared to normal controls. There was a negative correlation between levels of BDNF and the SLE disease activity index 2000 (SLEDAI-2K) (r=-0.349, P<0.0001). Depression (defined as BDI-II score≥18) was identified in 54 SLE patients (26.0%, 95%CI: 20%-31.9%). The serum BDNF levels were significantly lower in depression patients at the time of admission as compared with patients without depression [27.6(IQR, 23.2-30.4)ng/ml vs. 36.2(IQR, 31.7-42.3)ng/ml; P<0.0001]. Compared with the first quartile of serum BDNF levels, the second quartile OR for depression was 0.72 (95% CI, 0.61-0.80, P=0.033). For the third and fourth quartiles, it was 0.42 (95% CI, 0.33-0.52, P=0.002) and 0.16 (95% CI, 0.09-0.24; P<0.001). CONCLUSION: Serum BDNF levels are decreased in SLE patients with depressive symptoms. In SLE, serum BNDF levels are independently associated with depressive disorders, suggesting the role of neurotrophic factors in depression.

[Expression and Prognostic Significance of H3K27 Trimethylation Protein in DLBCL].

OBJECTIVE: To investigate the expression and prognostic effect of H3K27 trimethylation protein (H3K27me3) in diffuse large B-cell lymphoma(DLBCL). METHODS: A total of 102 DLBCL patients from Fujian Provincial Cancer Hospital were enrolled in this study. No therapy had been given before specimen collection. Tissue microarray(TMA) technique and immunohistochemistry(IHC) method were used for H3K27me3 immunostaining. Clinicopathologic and suvival data were carefully collected. The association between tested markers, clinicopathologic characteristics and prognosis were evaluated. Survival rates were analyzed by the Kaplan-Meier method, and prognostic factor were analyzed by the Cox proportional hazards model, the relation of different expression levels with clinical feature and prognosis of patients was compared. RESULTS: The quality of TMA was perfect and meet the standard of analysis. Among all DLBCL patients, 59.8% were characterized with high expression of H3K27me3, correlated with age, ECOG≥2, extranodular disease number≥2, elevation of LDH, medium-high risk IPI. Patients with high H3K27me3 expression manifested that the complete remission rate(CR) and overall remission rate (OR) were lower than those of patients with low expression, i.e., 20% vs 57.5% and 41.8% vs 90%, respectively (P<0.001). In addition, patients with high H3K27me3 expression showed shorter median survival time, i.e., 21.5 mon (P<0.0001). Multivariate analysis indicated that H3K27me3 was an independent risk factor for DLBCL patients (P=0.007). CONCLUSION: TMA technique is valid for the construction of DLBCL tissue chips. Patients with high expression of H3K27me3 indicates little response to treatment, worse outcome and shorter overall survival. The detection of H3K27me3 expression possesses a certain clinical value for prediction of DLBCL outcome.

[Ad-NK4 Enhances the Chemosensitivity of Human Multiple Myeloma RPMI 8226 Cells to Bortezomib].

OBJECTIVE: To investigate if Ad-NK4 can enhance the chemosensitivity of human multiple myeloma RPMI 8226 cells to bortezomib(BOR). METHODS: The cell-matrix adhesion test and PRMI 8226 cell-ECV 304 cell adhesion test were used to analyze the effect of Ad-NK4 on adhesion of RPMI 8226 cells; Western blot was used to detect the expression changes of adhesion and invasion-associated proteins MMP2, MMP3, MMP7 and VEGF; MTT assay was used to detect the effect of Ad-NK4 on proliferation of RPMI 8226 cells; the flow cytometry with PI staining was used to detect the effect of Ad-NK4 on cell apoptosis; the expression of cleaved caspase-3, BAX and BCL-2 was assayed by Western blot. RESULTS: These 2 adhesion assays indicated that Ad-NK4 significantly inhibited the adhesion of human multiple myeloma RPMI 8226 cells. In addition, Erk and JAK/STAT pathway may be involved in the process. The expression level of MMP-2, MMP-3 and VEGF were decreased in Ad-NK4 group, compared with untreated or Ad-GFP group (P<0.05). However, the expression of MMP-7 protein in Ad-NK4 group was not significantly different from untreated or Ad-GFP group (P>0.05). The inhibitory rates of the proliferation in cells treatedly Ad-NK4 combined with BOR was significantly higher than that with BOR or Ad-NK4 alone. Similarly, Western blot indicated that the level of cleaved caspase-3 and BAX in cells treated with Ad-NK4 combined with BOR was significantly higher than BOR or Ad-NK4 alone, but BCL-2 protein expression was significantly lower. Meanwhile, the ratio of BAX/BCL-2 was increased. CONCLUSION: Ad-NK4 can enhance the chemosensitivity of human multiple myeloma RPMI 8226 cells to BOR,which is associated with increasing of both BAX/BCL-2 ratio and Caspase-3 activation.

[Inhibitory effect of bone marrow mesenchymal stem cells on lymphoma cell proliferation].

Growing evidences show that mesenchymal stem cells (MSC) home to tumorgenesis and inhibit tumor cells, however, their molecular mechanisms are unclear. The purpose of this study was to explore the effect of B7-H4 in the influence of the mouse MSC on the proliferation of lymphoma cell line EL-4. The expression of B7-H4 on the MSC cell line C3H10T1/2 (C3H10) was detected by using immunofluorescence. FAM-siRNA was synthesized and transfected into C3H10 cells by INTERFER in (TM) siRNA Transfection Reagent. The transfection efficiency was determined by fluorescent microscopy and flow cytometry. The mRNA expression of B7-H4 was detected by RT-PCR after transfection of siRNA-B7-H4 into C3H10 cell line. The EL-4 was co-cultured with C3H10 siRNA-NC or C3H10 siRNA-B7-H4 for 48 hours, then was compared with EL-4 cultured alone, after 48 hours the cells were harvested under the confocal microscopy and measured by means of CCK-8 Kit. The results showed that the siRNA transfection efficiency in C3H10 cells reached to 72.43%, B7-H4 expressed highly on C3H10, the B7-H4 mRNA expression was down-regulated by transfection with different concentrations of siRNA into C3H10 cells. The proliferation of EL-4 was inhibited by C3H10 cells, and the effects were weakened and even disappeared after down-regulation of B7-H4. It is concluded that C3H10 can inhibit the proliferation of EL-4 through the expression of B7-H4, and this study provides new targets for the clinical treatment of lymphoma.

Knockdown of DEPTOR inhibits cell proliferation and increases chemosensitivity to melphalan in human multiple myeloma RPMI-8226 cells via inhibiting PI3K/AKT activity.

OBJECTIVE: The present study determined the role of DEP domain containing mTOR-interacting protein (DEPTOR) in the proliferation, apoptosis and chemosensitivity of RPMI-8226 multiple myeloma cells, using small hairpin RNA (shRNA) to knock down DEPTOR gene expression in vitro. METHODS: DEPTOR mRNA and protein levels in RPMI-8226 cells treated with DEPTOR-specific shRNA were evaluated by reverse transcription-polymerase chain reaction and Western blotting. Expression of apoptosis-associated proteins (including cleaved caspase-3 and cleaved poly-ADP ribose polymerase [PARP]) and activation of the phosphatidylinositol 3-kinase (PI3K)/v-akt murine thymoma viral oncogene homologue 1 (AKT) signalling pathway were detected by Western blotting. RESULTS: Transfection of DEPTOR-specific shRNA successfully knocked down DEPTOR gene expression in transfected RPMI-8226 cells. These transfected cells, together with control RPMI-8226 cells, were treated with 20 µmol/l melphalan for 24 h. Knockdown of DEPTOR exacerbated melphalan-induced growth inhibition and apoptosis, increased levels of cleaved caspase-3 and cleaved PARP, and reduced levels of phosphor-AKT. CONCLUSION: Downregulation of DEPTOR inhibited proliferation and increased chemosensitivity to melphalan in human multiple myeloma RPMI-8226 cells via inhibiting the PI3K/AKT pathway.

[Effect of DAPT on proliferation and apoptosis of human multiple myeloma cell line RPMI8226].

The aim of this study was to explore the effect of DAPT (N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycinet-butyl ester) on proliferation in vitro of human multiple myeloma cell line RPMI8226 and its underlying mechanism. The proliferation of RPMI8226 cells was detected by CCK-8 method; flow cytometry was employed to assay the cell apoptosis rate;the expressions of Notch1 and Hes1 proteins were detected by Western blot. The results indicated that the proliferation of human RPMI8226 cells significantly decreased after treatment with DAPT 0.5 - 5.0 µmol/L for 24 - 72 h (P < 0.05) in a concentration- and time-dependent manner. DAPT significantly induced apoptosis of RPMI8226 cells (P < 0.05). The expressions of Notch1 and Hes1 proteins were gradually downregulated with the increase of DAPT concentration. It is concluded that the DAPT can inhibit the proliferation of RPMI8226 cells, which may be related with the down-regulation of the protein expression of Notchl and Hes1.

[Calcium ionophore induces the differentiation of chronic myeloid leukemia cells into dendritic cells].

The aim of this study was to investigate the ability of calcium ionophore (CI ) to induce the differentiation of CML cells into dendritic cells (DC), to analyze the P210 expression in DCs and to evaluate the stimulatory effect of CML-DC on production of cytotoxic activity against CML cells via activating the autologous T cells. The mononuclear cells were isolated from bone marrow of CML patients whose WBC counts were more than 30x10(9)/L when samples were collected, then the lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI 1640 containing 10% FCS, with or without CI (375 ng/ml) and GM-CSF (200 ng/ml) at 37 degrees C, 5% CO2, fully humidified atmosphere for 96 hours. The cell morphology was observed under the inverted microscope and electron microscope; the expression of CD antigens was analyzed with flow cytometry; the P210 expression was measured with Western blot. LDH assay was used to evaluate the effect of cultured CML cells (CML-DC) generating cytotoxic T lymphocyte (CTL) activity against CML cells. The results indicated that after treatment with calcium ionophore and GM-CSF for 96 hours, CML cells showed DC morphological characteristics under inverted microscope and electron microscope. The expression of CD83, CD86, CD40, CD80 and HLA-DR increased remarkably. P210 was expressed in the CML-DC, but the expression level was lower than that in CML cells without CI and GM-CSF treatment. LDH assay showed that the CTL activity against CML was found greater in autologous T cells activated by CML-DC than that by CML cells. It is concluded that the CML cells can be induced to quickly differentiate into DC when cultured with CI and GM-CSF. CML-DC expresses P210, but the expression level is lower than that in CML cells. CML-DC can stimulate autologous T cells to produce CTL against CML.