【Objective】 To explore the therapeutic mechanismof arsenic trioxide(AS2O3),the active component of arsenolitumininhibitingtumor.【Methods】Humanlung adenocarcinoma cells(LAC) at logarithmic growth phase were culturedin vitro for 24 hours,and then were cocultured with arsenic trioxide(AS2O3) in the dosages of 0.75,1.0,1.5 and 2.0 mg/Lrespectively.Meanwhile,model control group was set up.The morphological features of LAC in different groups were observed under reverse microscope,growth-inhibitory rate of LAC was examined by methylthiazolyltetrazolium(MTT) assay,andthe expression of p53 gene was detected by enzyme-linked immunosorbent assay(ELISA) method.【Results】AS2O3in different dosages obviously inhibited the proliferation of LACin a time-effect manner,and markedly increased the expression of p53 gene(P

Objective To study the effects of extracellular-signal regulated kinase mitogenactivated protein kinase (ERK-MAPK) signaling pathway inhibition on histone phosphorylation and the related gene expression in human colorectal cancer cells.Methods Two human colorectal cancer cell lines (SW1116 and HCT116) were cultured and treated with gradient(0,20,40/μmol/L) doses of ERK-MAPK signaling pathway inhibitor U0126.Cell viability was determined by cell counting kit 8 (CCK-8) assay.Cell cycle distribution was assessed by flow cytometry.The expression levels of histone H3 kinases including ribosomal S6 serine-threonine kinase (RSK-2) and mitogen-and stressactivated protein kinase 1 and 2 (MSK1 and MSK2),and the levels of histone H3 (Ser10) phosphorylation and c-Fos protein were detected using Western blotting.Results Treatment of these two human colorectal cancer cell lines with ERK-MAPK inhibitor resulted in a time and dose-dependent inhibition of cell proliferation significantly. Proliferation rate of HCT116 was reduced to 47% at 72 hours after 40/μmol/L U0126 treatment. Cell cycle analysis showed that the percentage of phase G0/G1 cells significantly increased (P<0. 01) and the percentage of phase S cells decreased (P<0.01) after treatment with ERK-MAPK inhibitor. The expression of MSK1 and RSK2 reduced obviously in both of human colorectal cancer cell lines treated with U0126, which resulted in a 28% and 40% reduction of levels of MSK1 and RSK2 as compared with control HCT116 cells respectively,while no detectable change in the expression of MSK2 was found. Consistent with this, the expression level of histone H3 (ser10) phosphorylation was markedly down-regulated by ERK-MAPK inhibitor, and the related protein c-Fos expression decreased accordantly. Conclusions Decreased ERK-MAPK signaling pathway may reduce histone H3 (Ser10) phosphorylation via suppression of the activity of histone H3 kinase including MSK1 and RSK2, but not MSK2, consequently decrease the expression of c-Fos protein, which results in the inhibition of colorectal cancer cells proliferation.

Objective To investigate the effect of folic acid on the DNA methylation of tumorrelated genes promoters in healthy human peripheral blood mononuclear cells(PBMC). Methods Ten healthy volunteers were divided into two groups, and were randomized to receive either 5 mg folic acid (n=5)or placebo(n = 5) , one time per day for 3 months. The serum folic acid concentration was detected with chemiluminescence enzyme immunoassay kit before and after the intervention. The methylation statuses of five tumor-related genes promoter, including oncogenes c-myc, c-Ha-ras,tumor suppressor genes p16INK4A, E-cadherin and mismatch repair gene hMLH1 in PBMC were detected by bisufite sequencing. Results After folic acid intervention, the level of serum folic acid increased significantly in intervention group (t= -4. 739,P<0. 05) , however no significant difference in control group. After three-month folic acid intervention, the level of methylation of oncogene c-myc promoter increased from 4%, 3. 3%, 4. 1% before intervention, one week after intervention, one month after intervention respectively to 8%(t= -4. 079,P<0. 05), while no significant change in placebo taken group. Before and after the folic acid intervention, there was no significant difference of DNA methylation of other tumor-related genes promoter, including c-Ha-ras、E-cadherin、p16INK4Aand hMLH1. Conclusion Folic acid intervention can up-regulate DNA methylation of oncogene c-myc promoter, but can not affect the promoter methylation status of tumor suppressor genes E-cadherin,p16INK4Aand hMLH1.

Objective To investigate the effects of sinomenine on hind limb ischemia?reperfusion ( I∕R) injury and expression of Bcl?2 and Bax in skeletal muscle cells of rats. Methods Fifty?four healthy adult male Wistar rats, aged 6-8 weeks, weighing 180-220 g, were divided into 3 groups ( n=18 each) using a random number table: sham operation group ( group S) , group I∕R and sinomenine group ( group SIN) . The rats were subjected to 4 h of ischemia on the proximal part of the right hind limb using elastic rubber bands followed by reperfusion in I∕R and SIN groups. Sinomenine 60 mg∕kg was injected intraperito?neally at 30 min before reperfusion in group SIN, and the equal volume of normal saline was given instead of sinomenine at 30 min before reperfusion in S and I∕R groups. Immediately after onset of reperfusion and at 4 and 24 h of reperfusion, blood samples were collected from the cardiac apex to measure the concentra?tions of serum lactate dehydrogenase ( LDH) and creatine kinase ( CK) . The animals were sacrificed imme?diately after blood sampling, and the gastrocnemius specimens of the hind limb were immediately removed for determination of the wet to dry weight ratio ( W∕D ratio) and expression of Bcl?2 and Bax in gastrocnemi?us cells ( by immunohistochemistry) and for examination of the pathological changes after haematoxylin and eosin staining. The Bcl?2∕Bax ratio was calculated. Results Compared with group S, the gastrocnemius W∕D ratio and concentrations of serum LDH and CK were significantly increased, the expression of Bcl?2 was significantly down?regulated, the expression of Bax was significantly up?regulated, and the Bcl?2∕Bax
ratio was significantly decreased in I∕R and SIN groups ( P<0?05) . Compared with group I∕R, the gastroc?nemius W∕D ratio and concentrations of serum LDH and CK were significantly decreased, the expression of Bcl?2 was significantly up?regulated, the expression of Bax was significantly down?regulated, and the Bcl?2∕Bax ratio was significantly increased in group SIN ( P<0?05) . The pathological changes of the gastrocne?mius were significantly attenuated in group SIN as compared with group I∕R. Conclusion Sinomenine can attenuate hind limb I∕R injury, and the mechanism may be related to maintenance of the balance between Bcl?2 and Bax and to inhibition of apoptosis in skeletal muscle cells of rats.

To investigate the invasive ability of the residual tumor cells after immunotherapy and explore the feasible approach suppressing the invasion, mice were inoculated with B16 cells, and then treated by gene therapy with p4-1BBL/psPD-1 or IFN-gamma. The production and activities of MMP-9 and MMP-2 in residual tumor tissues were analyzed with gelatin zymography 1 day and 7 days after the termination of the immunotherapy. The production of MMP-9 and MMP-2 by B16 cells treated with IFN-gamma was also analyzed. IFN-gamma-treated B16 cells were inoculated to mice via subcutaneous injection. The invasion of tumor to muscular tissue was analyzed. Gene therapy with CH50 was used to suppress the invasive growth of tumor. The results showed that the expression and the activities of MMP-9 and MMP-2 were significantly increased 7 days after the end of immunotherapy. The response of tumor cells to ECM molecules was intensified after the removal of IFN-gamma, resulting in significant increase of both the production and activities of MMP-9 and MMP-2, and the increased invasion of tumor. Gene therapy with CH50 effectively suppressed the invasive growth of tumor. It is concluded that the termination of immunotherapy may result in a higher metastatic potential of residual tumor cells. Suppressing tumor invasion by suitable treatment will improve the efficacy of immunotherapy.

Objective To analyze clinical features and prognosis of primary intestinal T cell lymphoma (ITCL)which was misdiagnosed as Crohn′s disease (CD),and summarized the key points of differentiation between ITCL and CD.Methods From January 2003 to January 2014,clinical data of patients with ITCL once misdiagnosed as CD were retrospectively analyzed,which included demographic,clinical,pathological and prognostic data.The data of 177 patients diagnosed as CD from January 2012 to January 2014 were collected. The demographic,clinical,pathological and prognostic data of these two groups were analyzed and compared. The continuous variables were compared with t test or Mann-WhitneyU test,and the differences of classification variables between two groups were analyzed by Chi-square test or Fisher exact probability method.Results A
total of 18 patients (17 males and one female)with ITCL misdiagnosed as CD were enrolled in the study,and the median age at diagnosis was 38.5 (28.8 to 42.5)years and the median duration of diagnosis was 6.00 (3.75 to 13.25)months.The common primary symptoms were abdominal pain (12/18),diarrhea (13/18)and anemia (13/18).Intestinal perforation was primary symptom in two cases (2/18).However,B symptoms of lymphoma was observed in 16 patients,which included fever in 13 patients,weight loss in 16 patients and night sweat in one patient.One or more serious complications appeared in 12 patients,which included intestinal perforation in nine patients,severe gastrointestinal bleeding in seven patients and intestinal obstruction in two patients.In 177 patients with CD,104 patients were male (58.8%),and the median age at diagnosis was 22.0 (18.0 to 29.0) years.The primary symptoms were abdominal pain (88.7%,157/177),diarrhea (55.9%,99/177),anemia (63.8%,113/177),fever (33.3%,59/177)and weight loss (59.9%,106/177).During the disease course,30 patients (16.9%)had intestinal perforation (mainly chronic),12 patients (6.8%)had intestinal obstruction and seven patients (4.0%)had severe gastrointestinal bleeding.Compared to CD patients,male patients were more common in ITCL (χ2 =8.837,P <0.01),age at diagnosis was older (χ2 =314.5,P <0.01),the disease course was shorter (U=385.0,P <0.01),weight loss (χ2 =5.867,P <0.05)and fever (χ2 =10.609,P <0.01)were more common in clinical symptoms and intestinal perforation and severe gastrointestinal bleeding were more common in complications (χ2 =9.185,24.908,both P <0.01).The lesions of ITCL were multiple lesions, small bowel involved in eight cases,colon involved in 14 cases and one case with esophagus involved.Under endoscopy examination,most lesions appeared as ulcerations and were segmentally distributed.Compared to CD, lymphocyte proliferation was more common in the intestinal histopathological findings of ITCL (17/18 vs 19.7%(35/177);χ2 =42.844,P <0.01)and granuloma was rare (0 vs 42.8%(76/177),χ2 =12.665,P <0.01). Among 18 patients with ITCL,nine received chemotherapy and the median survival time was two months. Conclusions Primary ITCL had non-specific symptoms and was easily misdiagnosed as CD.More attention should be paid to the differential diagnosis of the two disease.

Objective To detect anti-M_2 autoantibody using recombinant BCOADC-E_2.Methods We purified recombinant BCOADC-E_2 by Ni~(2+)affinity chromatography column and then detect anti-M_2 autoan- tibody in the sera of patients with primary biliary cirrhosis(PBC)by Western blot test(WBT)and enzyme linked immunosorbent assays(ELISA).Results Among 60 sera from PBC patients,33 were positive,all of controls were negative.Conclusion The recombinant BCOADC-E_2 can be used to detect anti-M_2 autoanti- body specifically and sensitively.It is helpful for the diagnosis of PBC.

Objective To investigate the prevalence of uveitis associated with axial spondyloarthritis (ax-SpA), and analyze the features of patients with ax-SpA accompanying uveitis. Methods Two hundred and eighteen patients with ax-SpA were recruited. The differences in demographic factors, clinical features, the use of drugs and the quality of life were compared between uveitis group and non-uveitis group by using t-tests andχ2 tests. Results The prevalence of uveitis associated with ax-SpA was 24.3%. Uveitis group had longer disease duration [(156±140) month] compared to the non-uveitis group [(84±98) month] (t=-3.473, P=0.001), longer duration from disease onset to diagnosis [(90±105) month] compared to non-uveitis group [(47±65) month (t=-2.818, P=0.006), longer duration from first visit to diagnosis [(67±97) month] compared to non-uveitis group [(34 ±55) month] (t=-2.366, P=0.021). Patients with uveitis had higher rate of positive HLA-B27 (97.7%) compared to non-uveitis group (74.2%) (t=5.822, P=0.016), higher score of bath ankylosing spondylitis metrology index (BASMI) (3.3±2.0) compared to non-uveitis group (2.4±1.9) (t=-3.141, P=0.002), higher usage rate of biological agent (64.2%) compared to non-uveitis group (t=4.907, P=0.027). Conclusion In patients with acute anterior uveitis, the history should be carefully collected and HLA-B27 should be examined in order to make early diagnosis and treatment of ax-SpA, reducing uveitis flares and functional impairment.

Recent evidence has suggested that Akt2 plays an important role in the protection of cells from paclitaxel (PTX)-induced apoptosis and control of the cell cycle. In addition, some scholars suggested that the PTX sensitivity depends on a functional spindle assembly checkpoint. In the present study, we investigated the role of the Akt2/Bub1 cross-talking in apoptosis and cell cycle after exposure of the A2780 ovarian cancer cells to paclitaxel (PTX). Recombinant expression plasmid WT-Akt2 was transfected into A2780 cells by lipofectamine2000, and then the expression level of Akt2 gene was detected by using RT-PCR and Western blotting. Cell apoptosis and cell cycle were detected by flow cytometry and Hoechst 33342 staining after treatment with PTX. Moreover, we compared the expression level of Bub1 in different groups by Western blotting. Our study showed that up-regulation of Akt2 contributed to A2780 ovarian cancer cells overriding PTX-induced G(2)/M arrest, and inhibited Bub1 expression. Our findings might shed light on the molecular mechanism of PTX-induced resistance in ovarian cancer and help develop novel anti-neoplastic strategies.

AIM: To study the effect of ginkgolide B on high glucose-induced human lens epithelial cell (HLEC) apoptosis.
METHODS: The high glucose-induced HLEC model was established. Different concentrations of ginkgolide B were intervened. Cell viability was assayed by MTT assay, morphology of cell apoptosis was observed by hochest33258 staining. Cell ultrastructural changes were detected by transmission electron microscopy. Expressions of apoptosis - related factors caspses - 3 and caspase-9 were tested by colorimetric detection.
RESULTS: High glucose inhibited the activity of HLECs, induced apoptosis reaction of HLECs, caused high expression of apoptosis factors in cells; while ginkgolide B inhibited the decrease of cell viability induced by high glucose, decreased the HLEC apoptosis and reduced the expression of apoptosis factors Caspase-3 and Caspase-9.CONCLUSION: Ginkgolide B may inhibit the expression of caspses-3 and caspase-9 then effectively inhibited high glucose-induced apoptosis in HLECs.