AIM: To evaluate the relationship between uPA and NF-?B p65 expression and its clinical significance in breast cancer. METHODS: The uPA mRNA was measured by real-time fluorescent quantitative PCR in 46 cases of breast cancer tissues and their adjacent counterparts. NF-?B p65 were measured using immunohistochemistry. RESULTS: The expression of uPA gene was elevated in 63% of cases, and there was a strong correlation between NF-?B p65 and uPA expression (r=0.451,P

AIM:To observe the effect on NF-?B pathway and cell motility in breast cancer cell lines after transfection of dominant negative I?B? plasmid.METHODS:After stable transfection of mutant I?B? plasmid into highly metastatic breast cancer lines MDA-MB-231 and MDA-MB-435,we detected NF-?B binding activity by EMSA,cell growth ability by cell growth curve,colony forming test,and cell motility by millicell-PCF chamber.RESULTS:Constitutive activities in MDA-MB-231 and MDA-MB-435 were observed.Stable transfection of a dominant negative Ⅰ?B? resulted in downregulation of NF-?B binding activity,thus inhibited cell mobility without significant effect on cell growth.CONCLUSION:Cell migration ability is inhibited in highly invasive breast cancer cells by inhibition of NF-?B pathway in vitro.

Background Intracorneal macrophages play a critical role in corneal neovascularization (CNV)by secreting relative chemokines.But macrophages are characteristic by heterogeneity which has different biologic functions under different induction or stimulation from microenvironment.Objective This study was to detect the effects of chemokine (C-X3-C motif) ligand 1 (CX3CL1) and chemokine (C-C motif) ligand 2 (CCL2) on macrophages in vitro.Methods CNV was induced by corneal alkali burn in the left eyes of 20 male BALB/c mice aged 7-8 weeks.The CNV was evaluated under the slit lamp microscope 4 days after alkali burn,and then the corneal sections were prepared after mice were sacrificed.The expressions of CCR2 and CX3CR1 in the corneal specimens were detected by histo-fluorescence staining.Human peripheral blood mononuclear cells were separated using density gradient centrifugation and incubated in RPMI-1640 medium containing 10％ fetal bovine seruml(FBS) with 30 μg/L granulocyte-macrophage colony-stimulating factor (GM-CSF).The cells were divided into CD68 +CCR2 group and CD68+CX3CR1 group,and the percentage of the CX3CR1 and CCR2 expressions in the infiltrated macrophages of corneal specimens and human monocyte-derived macrophages was assayed by flow cytometry.The cultured cells were stimulated using human recombinant CX3CL1 and CCL2 proteins,and real-time PCR was used to detect the relative expressions of angiogenesis-related factors in macrophages.Results CNV was found in corneas 4 days after alkali burn and the CNV onsets from corneal limbus to central zone observed by a slit lamp.CCR2 and CX3CR1 were expressed in the F4/80-positive macrophages in alikali burned corneas.The macrophages grew for two weeks and appeared more dead cells in without GM-CSF group,but in GM-CSF induced group,the number of macrophages was increased.The percentage of CX3CR1-positive cells was 75％ and that of CCR2-positive cells was 45％.Real-time PCR showed that expression level of vascular endothelial growth factor (VEGF) mRNA increased and that ADAMTS-1 mRNA or TSP-1 mRNA decreased on macrophages after CCL2 stimulation,with significant differences in the 150 mg/L CCL2 group compared with the control group (t =-5.09,P =0.03 ; t =3.01,P =0.04 ; t =4.27,P =0.02).However,the VEGF mRNA expression decreased and ADAMTS-1 mRNA and TSP-1 mRNA increased after CX3CL1 stimulation,showing significant differences between the 150 mg/L CX3CL1 group and the control group (t=6.35,P=0.O2;t=-2.92,P=0.04; t=-3.81,P=0.03).Conclusions These results suggest that the macrophages have high heterogeneity.CCL2-and CX3CL1-expressing macrophages can regulate the expressions of angiogenesis-related factors.Macrophage chemokine signal may be a good target for treatment of neovascular ocular disease.

OBJECTIVE: To investigate the expression of periostin in in vitro cultured vascular smooth muscle cells (VSMCs) induced by TGF-β1 and the relationship between periostin expression and the migration and proliferation of the VSMCs. Further, to investigate the effects of atorvastatin on the above-mentioned processes and the molecular mechanisms of atorvastatin inhibition of TGF-β1- induced periostin production. METHODS: Rat aorta smooth muscle cells were cultivated by the method of tissue explants adherence. Cells of generation 3 to 6 were used as the experimental system. Primary cultured rat vascular smooth muscle cells were treated by TGF-β1 and different concentrations of atorvastatin,Y-2763 (Rho kinase inhibitor), or atorvastatin plus MVA for 24 hours. The expression of periostin was measured by RT-PCR and Western blot. A Boyden chamber assay was used to measure cell migration, and an MTT test was used to measure cell proliferation. RESULTS: Periostin expression in rat VSMCs stimulated by TGF-β1 increased significantly (4.158 ± 0.515 vs 0.385 ± 0.031), VSMC migration(25 ± 4 vs 8 ± 2) and proliferation (0.85 ± 0.06 vs 0.32 ± 0.03) also increased significantly. Atorvastatin significantly inhibited TGF-β1-induced periostin production in rat VSMCs, as well as VSMC migration and proliferation, in a dose-dependent manner. Rho kinase inhibitor Y-27632 significantly inhibited TGF-β1-induced periostin production in rat VSMCs (2.082 ± 0.245). The inhibitory effect of atorvastatin on periostin upregulation induced by TGF-β1 was reversed by mevalonate (3.838 ± 0.326). CONCLUSION Periostin can promote rat VSMC migration and proliferation. Atorvastatin inhibition of periostin expression induced by TGF-β1 in VSMCs may be exerted by inhibition of the production of MVA and other isoprene compounds and by blocking the Rho/Rho kinase signaling pathway.
Animals ; Atorvastatin ; Cell Adhesion Molecules ; genetics ; metabolism ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Heptanoic Acids ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Primary Cell Culture ; Pyrroles ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transforming Growth Factor beta1 ; pharmacology ; rho-Associated Kinases ; metabolism

AIM: To investigate the role of nuclear factor ?B (NF-?B) in the induction of IL-8 gene by TNF-? in colon cancer cells and the effect of antioxidant on the induction of IL-8. METHODS: ELISA was used to detect the concentrations of IL-8. IL-8 mRNA was analyzed by using RT-PCR. NF-?B in the cell nuclei was detected with electrophoretic mobility shift assay. RESULTS: (1) IL-8 production and IL-8 mRNA expression induced by TNF-? was blocked by pyrrolidine dithiocarbamate (PDTC). (2) TNF-? triggered the activation and translocation of NF-?B and PDTC inhibited the activation of NF-?B induced by TNF-?. CONCLUSION: The induction of IL-8 gene and protein by TNF-? is dependent on the activation of NF-?B. Antioxidants may inhibit the induction of IL-8 gene and protein through inhibiting NF-?B activation.

Inflammatory bowel disease(IBD),including ulcerative colitis(UC)and Crohn's disease(CD),is a group of chronic and relapsed inflammatory diseases of the intestinal tract. Genetic factors together with immunological, environmental and infectious factors contribute to the pathogenesis of IBD,however,the precise etiology is still uncertain. Susceptibility genes have great impact on the occurrence,development,clinical phenotypes,and even follow-up treatment of IBD,which indicates the significant roles of genetic factors in this disease. In this article,the well-studied and newly found genes related to IBD and their single nucleotide polymorphisms(SNPs)were reviewed.

Objective:The differential diagnosis of pulmonary artery sarcoma (PAS) and pulmonary embolism (PE) by double-detector spectral computerized tomography (CT) provides a new way to improve the detection rate of PAS and reduce the misdiagnosis rate.Methods:In the way of retrospective study, the Philips Nebula Workstation (ISP) was used to reconstruct electron density map, iodine density map and spectral curve in the spectral CT plain scan and enhancement of the PAS patient. In the plain scan image, the low density areas of the ascending aorta and the right pulmonary trunk were selected to measure their electronic density values. In the chest enhancement image, the iodine density of PAS area and PE area were measured respectively, and the spectral curves of PSA area and PE area were compared.Results:The electron density of the ascending aorta and the right pulmonary trunk in the low density area of the PAS patient during the plain scan of spectral CT were 104.4% EDW (relative to the percentage of the electron density of water) and 102.2% EDW, respectively, which were lower than those in the normal ascending aorta area. The fusion image of mixed energy and electron density clearly reflected the scope of the lesion. The iodine density in PAS area was 1.89 mg/ml, and the iodine density in PE area was 0.03 mg/ml during the enhancement phase. The iodine uptake in PAS area was significantly higher than that in PE area. The slope of PAS region was 2.08, and the slope of PE region was -1.86. The slopes of the two spectral curves were inconsistent.Conclusions:The electronic density, iodine density and spectral curve measured by double-detector spectral CT may provide powerful imaging basis for the diagnosis of PAS and the differentiation of PAS from PE, which is helpful for the early diagnosis of the lesions, and also provide basis for the biopsy location of the mixed lesions of PAS and PE.

Objective:To investigate the clinicopathological features and computed tomography (CT) findings of tracheal glomus tumor (GT) in order to improve the understanding and diagnosis of tracheal GT.Methods:The clinical and CT imaging data of 2 patients with tracheal GT diagnosed in the First Affiliated Hospital of Zhengzhou University were analyzed retrospectively. The image characteristics based on previous reports were analyzed.Results:The clinical manifestations of trachea GT were dyspnea, chest tightness, hemoptysis, etc., which were easy to be misdiagnosed. The CT manifestations were spherical or nodular protrusions in the trachea cavity, with uneven edges, which can be lobulated. Cystic changes can be seen in the focus. After enhancement, it showed progressive filling and obvious enhancement, without deep infiltration and distant metastasis.Conclusions:Chest CT can accurately localize tracheal GT, provide its morphological size, blood supply, growth characteristics and other characteristics, accurately display the overall morphology of the lesion, and provide some help for the development of the surgical plan, and its definitive diagnosis still relies on pathological examination.

Objective:To understand the clinical features and computed tomography (CT) imaging performance of mediastinal giant malignant synovial sarcoma (SS) and to improve the clinicians′ diagnosis and treatment of this disease.Methods:We report the clinical data of 2 cases of primary mediastinal giant malignant SS, and reviews the literature for CT presentation and differential diagnosis.Results:Primary giant malignant SS of mediastinum is rare in clinic. CT showed irregular cystic and solid mass with mural nodules, adjacent osteolytic destruction, unclear boundary with adjacent tissues. The enhancement showed that the solid components of the tumor and mural nodules were enhanced, while the cystic necrosis area had no enhanced uneven enhancement. The two cases in this paper showed " borehole-like" growth above the clavicle and growth into the left thoracic cavity, with pulmonary metastasis and lymphatic metastasis respectively.Conclusions:Mediastinal malignant SS is a rare disease with high malignancy and easy to metastasize, which emphasizes " early detection, early diagnosis and early treatment" . It should be taken into account in the differential diagnosis of mediastinal tumors, but the confirmation of diagnosis needs to be combined with pathological and immunohistochemical findings. Clinicians need to take advantage of imaging examinations to provide auxiliary guidance on the diagnosis, metastasis assessment and surgical treatment options of SS.

Objective To observe the effects of Shengxue Tongbian Granules on colonic myoelectricity and Ca2+/CaM/MLCK signaling pathway in rats with slow transit constipation(STC)of blood deficiency and intestinal dryness syndrome;To explore its mechanism for the treatment of STC.Methods The STC model of blood deficiency and intestinal dryness syndrome was established by intragastric administration of loperamide combined with tail bloodletting.The rats were divided into control group,model group,mosapride citrate group and Shengxue Tongbian Granules group,with 8 rats in each group.The administration group was given corresponding drugs by gavage for 14 days.The general condition of rats before and after treatment was observed,the fecal water content was detected,the slow wave frequency,amplitude,and coefficient of variation of colonic electromyography were detected using a biological function experimental system,and intestinal propulsion rate was detected.HE staining was used to observe the pathological changes of colon tissue,the concentration of Ca2+ in colonic smooth muscle cells was detected by colorimetry,the expression of Cx43,calmodulin(CaM),myosin light chain kinase(MLCK)and p-MLC20 in smooth muscle tissue were detected by Western blot.Results Compared with the control group,the body mass,fecal water content and intestinal propulsion rate of rats in the model group decreased(P<0.01),the slow wave frequency of colonic electromyography slowed down,the coefficient of variation of frequency increased(P<0.01),and the amplitude and coefficient of variation of slow wave increased(P<0.01);colonic mucosal structure was damaged,with visible inflammatory changes and significant erosion,and the concentration of Ca2+ and the expressions of Cx43,CaM,MLCK,p-MLC20 proteins in colonic smooth muscle cells were significantly decreased(P<0.01).Compared with the model group,the body mass,fecal water content and intestinal propulsion rate of the rats in the mosapride citrate group and the Shengxue Tongbian Granules group significantly increased(P<0.05,P<0.01),the slow wave frequency of colonic electromyography increased and the coefficient of variation of frequency decreased(P<0.01),and the slow wave amplitude and amplitude variation coefficient decreased(P<0.05,P<0.01);the colonic mucosal structure was relatively intact,the erosion situation was improved,and the Ca2+ concentration,Cx43,CaM,MLCK and p-MLC20 protein expressions in colonic smooth muscle cells significantly increased(P<0.01).Conclusion Shengxue Tongbian Granules can improve defecation symptoms and promote colonic motility in STC rats with blood deficiency and intestinal dryness syndrome,which may be related to regulating colonic myoelectric rhythm and activating Ca2+/CaM/MLCK signaling pathway.