AIM:To investigate the effects of survivin inhibitor YM155 {4,9-dihydro-1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(2-pyrazinylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide} on the apoptosis, mito-chondrial membrane potential (Δψm) and cytochrome C (Cyt C) of retinoblastoma Y79 cells, and to analyze the mitochon-drial mechanisms of apoptosis .METHODS:Y79 cells were cultured in vitro and treated with YM155 at concentrations of 0, 0.5, 1, 2, 4 and 8 nmol/L.The cells in control group were treated without YM 155.The proliferation of Y79 cells were measured by CCK-8 assay and bromodeoxyuridine ( BrdU) labeling assay .Y79 cells were randomly divided into 4 groups:control group ( with equal volume of RPMI-1640 nutrient medium ) , positive control group ( 10 nmol/L topotecan ) , low-dose (1 nmol/L) YM155 group and high-dose (2 nmol/L) YM155 group.The effects of YM155 on the apoptosis, the changes of Δψm , the mitochondrial distribution and the protein level of Cyt C in the Y 79 cells were evaluated by flow cytom-etry with Annexin V-FITC/PI staining, JC-1 staining, immunofluorescence analysis and Western blot , respectively.RE-SULTS:Compared with control group , YM155 significantly inhibited the proliferation of Y 79 cells and induced apoptosis (P<0.05).YM155 significantly reduced Δψm of the Y79 cells, promoted Cyt C which released from mitochondria to the cytosol and reduced the protein level of Cyt C in the mitochondria (P<0.05).CONCLUSION:YM155 inhibits Y79 cell proliferation and induces apoptosis , and the possible mechanisms may be involved in the mitochondrium-mediated apoptotic pathway .

Objective To detect the changes in the expression of discoidin domain receptor 2(DDR2)and matrix metalloproteinase (MMP)-13 in different stages of cartilage and synovium damage of osteoarthritis rats.The relation between DDR2 and the degree of cartilage damage was explored.Methods Modified papain knee joint injection approach was adopted to establish animal model of OA.The expression and distribution of protein of DDR2 and MMP-13 were checked in articular cartilage and synovium at different stages of OA.Results The expressions of DDR2 in articular cartilage and synovium of experimental groups were different from those of the normal group (P＜0.01).They were higher in cartilage than those in the corresponding synovium.The expressions of MMP-13 demonstrated the same characteristics with those of DDR2,r=0.93(P＜0.01).Conclusions The important role of DDR2-MMP-13 in cartilage damage has been proven in the pathogenic process of OA.The upregulated expressions of DDR2 in articular cartilage and synovium have a detrimental effect on cartilage degeneration.

Objective To study the anti-fibrotic function and mechanism of peroxisome proliferator activated receptorγ(PPARγ) in connective tissue disease-interstitial lung disease (CTD-ILD).Methods The expression of PPARγin lungs was analyzed in 37 cases with CTD-ILD and 20 control cases by immunohistochemistry.Changes in α-SMA levels were analyzed by Western blotting,and acetylation of Smad3 and Smad3 or PPARγ combined with P300 were analyzed by IP-WB.The data was analyzed by one-way ANOVA,t test or Mann-Whitney test.Results PPARγ' expression in the lung of CTD-ILD was lower than the controls [0.92％(1.44％),3.50％(1.94)％,respectively; Z=-8.924,P＜0.01].Different concentration of PPARγ (0,1,5,10,20,40 pmol/L) ligandinhibited the marked elevation of the protein α-SMA induced by TGF-β1 in a concentration-dependent manner (0.918 ±0.062,0.852±0.042,0.725 ±0.057,0.678 ±0.042,0.418 ±0.022,0.456±0.029; P＜0.05 or P＜0.01).However,this response was blocked by a selective antagonist PPARγ signaling GW9662 (0.946±0.087 vs 0.538±0.120,P＜0.01).Acetylation of Smad3 expression was increased when TGF-β1 was putted into lung fibroblasts after 60,90 and 180 min (0.565±0.047,1.127±0.101,0.873±0.022,0.614±0.407; all P＜0.05).The combination of Smad3 with P300 was also increased (1.46±0.12,0.98±0.09; P＜0.05),compared with the controls.But the ligand of PPARγ could block this effect (0.62±0.10,1.46±0.12; P＜0.05).Meanwhile,the combination of PPARγ and P300 was increased (0.94±0.05,0.76±0.22; P＜0.05).Conclusion PPARγ may play a physiologic role in the regulation of anti-fibrosis response.Its function may be realized by its competition with Smad3 combined with P300.

AIM:To investigate the effects of gonadotropin-releasing hormone ( GnRH) analogue on the growth of breast cancer cell lines MCF-7 and MDA-MB-231 in vitro and to explore the related mechanisms with PI 3K/Akt or ERK/MAPK pathways .METHODS: The proliferation of human breast cancer cell lines MCF-7 and MDA-MB-231 treatment with triptorelin was detected by MTT assay and the distribution of the cell cycle was determined by flow cytometry .The phosphorylation of the ERK 1/2 and Akt was evaluated by Western blotting .RESULTS:Triptorelin inhibited the prolifera-tion of MCF-7 cells at concentration of 10-5 mol/L after treated for 192 h or at concentration of 10 -4 mol/L after treated for 168 h and 192 h.Triptorelin inhibited the proliferation of MDA-MB-231 cells at concentration of 10 -4 mol/L after treated for 192 h (P<0.05).Treatment with triptorelin for 192 h at concentration of 10 -4 mol/L had no statistical significance effect on phosphorylation of ERK1/2 and Akt(P>0.05).CONCLUSION:Inhibitory effect of GnRH analogue triptorelin on human breast cancer cells is not just the connection with the down-regulation of pituitary hormone , but also a direct in-hibitory effect.The role may not be involved in the activation of ERK /MAPK and PI3K/Akt signaling pathways .

Objective To study the pro-invasive effect of irradiation on human glioblastoma cell line U87 and its possible mechanism.Methods Cultured U87 cells received different doses of irradiation (0,2,and 4 Gy).The change in cellular invasiveness was measured using the real-time cell analyzer system.The activities of matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9) in U87 ceils were measured by gelatin zymography before and after irradiation.The content and distribution of intracellular β-catenin after irradiation were determined by immunohistochemistry.The mRNA levels of Wnt/β-catenin target genes were measured by real-time quantitative PCR.Results After irradiation,the invasiveness of U87 cells increased significantly (P ＜ 0.01),which was dose-dependent within a certain dose range; the activities of MMP2 and MMP9 in U87 cells increased significantly (P =0.031 for MMP2 ; P =0.004 for MMP9) ;the content of β-catenin in U87 cells increased significantly (P ＜ 0.01),with translocation from the cell membrane and adherens junctions to the nucleus; the mRNA levels of Wnt/β-catenin-related genes (FZD7 and TCF1) increased significantly (P ＜ 0.01),and the transcription of Wnt/β-catenin target genes,especially those related to migration and invasion such as MMP2,MMP7,MMP9,and CD44,was significantly enhanced (P ＜ 0.05).Conclusions Irradiation can promote the invasion of glioblastoma U87 cells,possibly by activating the Wnt/β-catenin pathway and enhancing the transcription of migration-and invasion-related genes.

Objective:To investigate the strategies and methods of pancreatic cancer immunotherapy adopt cytotoxic T lymphocytes activated by dendritic cells(DCs),which modified by heat shock protein 70 peptide complex(HSP70-PC).Methods:HSP70 peptide complex from lumps of tumor-bearing mouse innoculated with pancreas cancer cell(MPC83)were purified by ConA-Sepharose and ADP-Agarose affinity chromatography.Dendritic cells derived from normal murine bone marrow were induced by GM-CSF and IL-4.MTT method was applied to analyses the proliferation activities of DCs.To achieve the specific T lymphocyte cells,DCs modified with HSP70 peptide complex were coculture with murine spleen lymphocytes for 48 hours.Then these activated lymphocytes were cocultured with MPC83 at ratio of effect-target 10∶1 and 20∶1 in vitro and the killing activities were detected by MTT method.Results:High purify HSP70-PC was obtained;104 DCs need 50～100ng of HSP70-PC to modify,100?g of HSP70-PC peptide complex can be extracted from per 1g lump;T cells stimulated by HSP70-PC modify DCs showed more specific cytotoxic activity to MPC83 than lymphocytes activated by DCs without HSP70-PC in vitro.Conclusion:We can extract high purity HSP70-PC for clinical individual immunotherapy from pancreatic cancer lump by using hypobaric affinity chromatography;DCs vaccine modified with HSP70-PC has significant kill effect against pancreatic cancer cell in vitro.

Objective To investigate the expression of AIB1 protein in bladder cancer and its significance.Methods 30 cases of bladder cancer and 30 adjacent normal tissues were collected from 30 patients underwent radical or TURBt.The level of AIB1 protein expressed in these tissues were detected by immunohitochemical method.Results The positive rate of AIB1 protein in bladder carcinoma was higher than that in the normal tissues [43.3 ％ (13/30) vs 3.3 ％ (1/30),x2 =4.316,P ＜ 0.05].The positive rate of AIB1 in T3 stage was higher than that in T1 or T2 stage [62.5 ％ (5/8) vs 25.0 ％ (2/8),42.9 ％ (6/14),x2 =4.623,P ＜ 0.05].No correlation was found between the expression of AIB1 protein and the sex,age,multiplicity or pathological grade (P ＞ 0.05).Conclusions There is a close relationship between AIB1 and bladder cancer.AIB1 might have great value on the prognosis of patients with bladder cancer.

With modified mutual information(MI) measure,Powell search algorithm is utilized to realize mono-modality image registration.The MI's formulae are improved to reduce computation cost,and the replace of Powell's search direction is also rectified to keep the original search direction and eliminate local maxima.With rotation set in advance,local maxima can be avoided.The influence on registration velocity by Powell convergence threshold is also discussed.The results show that the modified MI measure can reach sub-pixel precision with obviously accelerated speed.It has good robustness,high speed and high precision.

Objective:To summarize the perioperative nursing experience of a patient with hemophilia arthritis complicated with hepatitis C undergoing unilateral hip and knee arthroplasty at the same time.Methods:In August 2018, a patient with hemophilic arthritis complicated with hepatitis C was admitted to the first hospital of Lanzhou University. Preparations for health education, functional exercises, examination and evaluation before operation was made. Unilateral hip and knee arthroplasty were performed at the same time, strengthen the management of incisions and catheters after the operation, actively prevent complications, and guide patients to perform functional rehabilitation exercises under the conditions of good pain management and safe coagulation factor activity; pay attention to the nursing of coagulation factor replacement treatment in the whole perioperative period.Results:After careful nursing, no serious complications occurred during the patient's hospitalization, and the position and force line of the prosthesis were satisfactory by imaging examination before discharge.Conclusion:The perioperative care of this patient proved that the above-mentioned nursing measures are effective for hemophilic arthritis patients with hepatitis C undergoing unilateral hip and knee arthroplasty at the same time.

Objective To study the relationship between nasopharyngeal carcinoma and positive IgA antibodies of EB virus nuclear antigen 1 (NA),EB virus capsular antigen (VCA) and EB virus Zta protein.Methods The serum EB virus VCA-IgA,NA1-IgA and Zta-IgA antibody in 17 175 cases were detected by ELISA.782 cases of EB virus antibody-positive subjects were further given nasopharyngeal CT imaging,electronic nasopharyngoscopy.Finally confirmed by biopsy and immunohistochemistry.Comparison of serology results EB antibody-positive individual,while the two antibody-positive and antibody while three positive nasopharyngeal carcinoma detection rate,combined with evaluation of EB virus antibody detection in nasopharyngeal carcinoma screening value of high-risk groups.Results 17 175 cases of medical groups,the EB virus antibodies were detected in 782 cases,with the most positive individual,accounting for 535 cases (68.41％),nasopharyngeal cancer diagnosed in 15 cases (2.80％);two antibody positive while 213 cases (27.24 ％),diagnosed 49 cases of nasopharyngeal carcinoma (23.00％);least three antibody positive while only 34 cases (4.35％),nasopharyngeal cancer diagnosed 18 patients (52.94％);by physical examination,in 16,393 cases of EB virus antibody negative population by nasopharyngoscopy and throat CT and other medical examination,the final diagnosis of nasopharyngeal carcinoma in 6 cases (0.04％).Conclusion Three EB virus antibody combined detection greatly improves the rate of nasopharyngeal cancer diagnosis;we must also do other physical examination to prevent EB virus antibody-negative nasopharyngeal crowd missed.