Objective To investigate the effect and mechanism of action in pancreatic cancer BxPC3 cell treated by cerulenin in combination with gemcitabine.Methods BxPC3 cells were cultivated with 10 μg/ml cerulenin,20 μmol/L gemcitabine or 10 μg/ml cerulenin + 20 μmol/L gemcitabine for 48 h,and cells without treatment were control.Cell proliferation was detected by CCK-8 assay,and early apoptosis was detected by AnnexinV/PI double staining method.The expression of Bcl-2 mRNA and Bax mRNA were detected by RT-PCR,and the protein level of Bcl-2,Bax were detected by Western Blot.Results The inhibition rate of BxPC3 cells were 0,(51.28 ± 1.84) ％,(53.59 ± 1.62) ％,(84.57 ± 1.01) ％ in control,cerulenin group,gemcitabine group,combination group; and the rate of early apoptosis was (0.83 ± 0.31) ％,(31.37 ± 1.04) ％,(38.33 ± 0.75) ％,(69.43 ± 0.83) ％,and the expression of Bcl-2 mRNA was 0.67 ± 0.01,0.44 ±0.01,0.36 ±0.08,0.27 ±0.07,and the expression of Bax mRNA was 0.14 ±0.01,0.31 ± 0.02,0.32 ± 0.03,0.91 ± 0.06 ; while the ratio of Bcl-2 mRNA/Bax mRNA was 4.78 ± 0.13,1.39 ± 0.04,1.15 ± 0.02,0.30 ± 0.02 ; the expression of Bcl-2 protein was 1.24 ± 0.04,0.51 ± 0.02,0.42 ± 0.02,0.13 ±0.01 ; and the expression of Bax protein was 0.20 ± 0.05,0.47 ± 0.01,0.54 ± 0.01,1.21 ± 0.03 ; while the ratio of Bcl-2/Bax was 6.00 ± 0.11,1.11 ± 0.01,0.77 ± 0.03,0.10 ± 0.06.And the difference among the groups was statistically significant (P ＜ 0.01).Conclusions Cerulenin combined with gemcitabin has synergistic effect on the inhibition of BxPC cells proliferation.Its mechanisms may be up-regulation of Bax,down-regulation of Bcl-2,and promoting apoptosis of cells through the decrease of Bcl-2/Bax ratio.

ObjectiveTo investigate the effect of thalidomide on the proliferation and apoptosis in human pancreatic cancer cell SW1990 in vitro.MethodsSW1990 cell line was treated with thalidomide at different concentrations (3.125,6.25,12.5,25,50,100,200 and 400 μg/ml) for 24,48,72 h,and then cell proliferation were evaluated by MTT.Cell cycle was determined by flow cytometry,apoptosis was determined by annexin V/PI fluos staining,Bcl-2,Bax protein expression and Bcl-2/Bax ratios were measured by Western Blot in vitro.ResultsThalidomide inhibited the proliferation of SW1990 cells in a time and dosedependant manner.The proportion of G0/G1 phase of SW1990 cells with 200 μg/ml thalidomide treatment increased from (41.15 ± 2.23 ) ％ to (58.83 ± 2.33 ) ％,apoptosis rate increased from 2.6％ to 28.0％,the expression of Bax protein up-regulated from 0.17 ± 0.03 to 0.33 ± 0.04,the expression of Bcl-2 protein downregulated from 0.35 ± 0.02 to 0.17± 0.01,the ration of Bcl-2/Bax decreased from 2.17 ± 0.44 to 0.52 ±0.07.ConclusionsThalidomide can inhibit the proliferation of pancreatic cancer SW1990 cells by upregulating Bax,down-regulating Bcl-2,inducing cell apoptosis and cell Go/G1 phase arrest.

Objective To observe the clinical effect of Shuguanling(Chinese medicine for dredging the oviduct) on salpingemphraxis sterility and to study its mechanism.Methods The diagnosed patients were randomized into 3 groups with 52 in each.The treatment group was administered Shuguanling p.o.and clyster.The control group A was given Xuefu Zhuyu Oral Liquid(Oral liquid for removing blood stasis in the abdomen),and Kangfu Xiaoyan Pills(Pills for recovery of gynecological inflammation) suppository into recta.The control group B was prescribed Xuefu Zhuyu Oral Liquid and vaginal culdocentesis with the mixture of dexamethasone,chymotrypsin,gentamicin and normal saline.After three months,the effect was evaluated and the pelvic blood flow,blood viscosity,plasma collagenase,fibrinogen and fibrinolytic activity of the treatment group were tested for comparison.Results The tubal patency rate and pregnancy rate of treatment group were significantly higher than those of control groups(P

Objective To evaluate the significace of endoscopic diagnosis of gastrointestinal graft-versus-host disease ( GI GVHD) resulted from allogeneic bone marrow transplantation. Methods Five patients with suspected GI GVHD received endoscopic examination and biopsies were taken from antrum, sig-moid colon or the focal lesions. Results The symptoms of GI GVHD included anorexia, nausea, vomiting, watery diarrhea, abdominal pain, GI bleeding etc. The endoscopic appearance in stomach varied from subtle mucosal edema, hyperemia, and erythema necrotic erosion. Besides the above stated, the endoscopic appearance in colon also presented with diffuse mucosal erosion, hemorrhege and ulcer. Histological findings included characteristically crypt epithelial cell apoptosis and sloughing, and lymphocyte infiltration in epithelium and lamina propria. The involvement may be varied from diffuse to focal in stomach or colon. Conclusion Endoscopic and histological evaluation of the stomach and colon and biopsy specimen can be used as the evidences in diagnosing GI GVHD.

Objective To explore the role of receptor‐interacting protein kinase 3 (RIP3)‐mediated necroptosis in ulcerative colitis (UC ) . Methods The colonic mucosa tissues of eighty‐five patients diagnosed with UC were collected .Disease staging and pathological classification of UC patients were evaluated according to Mayo standard and Truelove standard ,respectively .The expressions of RIP3 and tumor necrosis factor (TNF)‐α were detected by immunohistochemistry .The cell inflammation model in HT‐29 cells was induced by interferon (IFN)‐γ.The expression of RIP3 mRNA and interleukin (IL)‐1βmRNA was determined by real time polymerase chain reaction (RT‐PCR) .Twenty healthy individuals were studied as controls .A single factor analysis of variance was performed in the comparison among groups ,and Pearson correlation analysis was used for correlation analysis .Results Among 85 patients with UC , 27 cases were mild ,30 cases were moderate and 28 cases were severe;for pathological classification ,26 were gradeⅠ ,31 were gradeⅡ and 28 were grade Ⅲ .The Mayo score was positively correlated with pathological grade (r=0 .997 , P<0 .05) .The expression of RIP3 and TNF‐α in colonic mucosa tissues of UC group increased along with the pathological classification grade .The expression of RIP3 of gradeⅠto Ⅲ was 1 .81 ± 0 .98 ,2 .77 ± 1 .26 and 4 .86 ± 2 .77 ,respectively ,and the differences were statistically significant (F=26 .10 ,P<0 .05) .The expression of TNF‐αof gradeⅠto Ⅲ was 1 .35 ± 0 .69 ,2 .61 ± 1 .41 and 4 .43 ± 2 .17 ,respectively ,and the differences were statistically significant (F=31 .80 ,P<0 .05) .The expression of IL‐1β mRNA of IFN‐γ group ,IFN‐γ+ z‐VAD group and IFN‐γ+z‐VAD+ TNF‐αgroup was 0 .68 ± 0 .27 ,1 .47 ± 0 .12 and 1 .86 ± 0 .16 ,respectively ,and the differences were statistically significant (F=38 .45 , P<0 .05) .The expression of RIP3 mRNA of IFN‐γgroup ,IFN‐γ+z‐VAD group and IFN‐γ+z‐VAD+TNF‐αgroup was 0 .46 ± 0 .13 ,1 .21 ± 0 .29 and 2 .06 ± 0 .20 ,respectively , and the differences were statistically significant (F= 55 .15 , P< 0 .05) .Conclusions TNF‐α/RIP3 signal pathway is up‐regulated in UC .RIP3‐mediated necroptosis promotes the death of colonic epithelial cells and exacerbates inflammatory response .RIP3‐mediated necroptosis may be an important mechanism in UC .

Objective To study on the endoscopic, pathologic and immunohistochemical characteristics of gastric stromal tumor. Methods The gastric tumor samples were studied with light microscopy, and in those suspected of mesenchymal tumor the expression of CD11734, CD34 , vimentin, desmin, S-100 etc. were further determined with SP method. The clinical, pathological and endoscopic characteristics were reviewed. Results Twenty-eight cases of mesenchymal cell tumor were diagnosed as stromal tumor and enrolled in this study, with 9 benign, 11 borderline, and 8 malignant cases. Endoscopically, the gastric stromal tumors showed submucosal protuberant lesions, and were situated in fundus (15 cases), body (10 cases) and an-trum (3 cases). Deep concave ulcer presented on tumor surface in 10 patients. Histology revealed the typical stromal tumor features with cell in spindle and/or epithelia shape and arranged in fence-like or spiral forms. All cases were positive for CD117 CD34 , vimentin when assessed, but 28 cases were negative for desmin and 22 cases (18. 2% ) for S-100. Conclusion The gastric stromal tumors were the most common mesenchymal tumors. Microscopically, the tumors were composed of spindle or epithelioid cells. CD117 and CD34 can be used as immune markers to diagnose gastric stromal tumor.

Objective To identify a high affinity IRF-1 binding element in the translational start site of XAF1 promoter. Methods By the bioinformatics analysis, a putative IFN regulatory factor 1 binding element (IRF-E), named as IRFE-XAF1, was identified from -30nt to -38nt of the XAF1 gene, with 76.2% homogeneity with the synonymous IRF-E sequence. Electrophoresis mobility shift assay (EMSA) was performed to confirm the binding capacity of IRFE-XAF1. Two site-directed mutations were made, one mutation site was outside of the IRF-E region (-28nt) and another located at the center of IRF-E (-34nt). The promoter of XAF1 after mutation was examined, including its binding activity and response to IFN-?. Results It was found by EMSA assay that the doublestranded oligonucleotide DNA probe, containing IRFE-XAF1 and labeled with 32P, may be connected to the nuclear protein, and blocked by the unlabeled synonymous IRF-1 probe (cold probe). The binding capacity of IRF-E was lost after site-directed mutation. The XAF1 promoter containing IRFE-XAF1 site had the priming activity, and could be induced by IFN-?. The priming activity declined markedly after site-directed mutation of IRFE-XAF1, and -34 site mutation completely eliminated the effect of IFN-?. Conclusion A high affinity of IRF-E is found in -30nt to -38nt region upstream of ATG initiator codon of XAF1 gene. The code sequence is -38nt-GAAACGAAA--30nt. The present study suggests that XAF1 is one of the genes with which IFN-? may induce the differentiation of cancer cells.

Objective To study the early diagnostic value of CT for children with cerebral palsy.Methods CT manifestations of 124 cases with cerebral palsy were analysed retrospectively. Results 96 cases of the 124 cases showed a recognisable abnormality,the abnormal rate of CT was about 77.42％ ,and the most common abnormality was cerebral atrophy.Spastic type was the most common clinical type(71.77％ ).The less of the age,the higher of the abnormal rate of CT. Conclusion Although CT isn't the main basis of the diagnosis of cerebral palsy,it's helpful for us to find the pathological changes,and to find the eliology and location.It also provided basis for us to judge the prognosis of cerebral palsy.So CT has significant value in the early diagnosis of cerebral palsy.

To evaluate the feasibility of using polyethyleneimine (PEI) coated magnetic iron.oxide nanoparticles (polyMAG-1000) as gene vectors. The surface characteristics of the nanoparticles were observed with scanning electron microscopy. The ability of the nanoparticles to combine with and protect DNA was investigated at different PH values after polyMAG-1000 and DNA were combined in different ratios. The nanoparticles were tested as gene vectors with in vitro transfection models. Under the scanning electron microscope the nanoparticles were about 100 nm in diameter. The nanoparticles could bind and condense DNA under acid, neutral and alkaline conditions, and they could transfer genes into cells and express green fluorescent proteins (GFP). The transfection efficiency was highest (51%) when the ratio of nanoparticles to DNA was 1:1 (v:w). In that ratio, the difference in transfection efficiency was marked depending on whether a magnetic field was present or not: about 10% when it was absent but 51% when it was present. The magnetic iron oxide nanoparticles coated with PEI may potentially be used as gene vectors.
Breast Neoplasms/metabolism ; Breast Neoplasms/*pathology ; Cell Line, Tumor ; Ferric Compounds/*chemistry ; Ferric Compounds/metabolism ; Gene Targeting ; Genetic Vectors ; Green Fluorescent Proteins ; Magnetics ; Nanotechnology ; Particle Size ; Polyethyleneimine/chemistry ; Transfection/methods

The aim of this study was to evaluate the feasibility and efficacy of using TRAIL gene to treat breast cancer mediated with a novel carrier - magnetic iron oxide nanoparticles (poly-MAG-1000) coated with PEI. The magnetic iron oxide nanoparticles were used as gene carrier to transfect TRAIL gene into MCF-7 cells. The polyMAG-1000 without TRAIL gene was transfected into the tumor cells as negative control. TRAIL gene transfection with liposome as carrier served as positive control. The apoptosis of cells was detected with TUNEL method. The apoptosis ratio of tumor cells was measured with flow cytometry (FCM). It was found that the apoptosis occurred in the tumor cells after transfection of TRAIL gene mediated by both polyMAG-1000 and liposome. The apoptosis ratio in the group with polyMAG-1000 as gene carrier was (25.11+/-2.85) %, whereas it was (5.06+/- 1.05) % in the control group with polyMAG-1000 (P<0.01). The apoptosis ratio was as low as (18.31+/-2.44) % in the group with liposome as gene carrier (P<0.05, as compared with the group with polyMAG-1000 as gene carrier). It is suggested that TRAIL gene may induce apoptosis in MCF-7 breast cancer cells. The magnetic iron oxide nanoparticles coated with PEI may be a potential gene carrier with high transfection efficacy for cancer gene therapy..