Objective: To dynamically evaluate left ventricular perfusion, global and local functional changes during left ventricular aneurysm (LVA) formation and to explore the relationship between the size of LVA and LVEF, LVESV, LVEDV by gated99mTc-MIBI SPECT (GSPECT) and gated18F-FDG PET metabolic (GPET) imaging in experimental pigs. Methods: LVA model was established by occlusion of left circumlfex artery (LCX) and placing an Ameroid constrictor at the proximal end of left anterior descending artery (LAD) in a total of 16 Chinese mini-pigs. At the 1st, 4th and 8th weeks of surgery, the changes of total perfusion defect (TPD), LVA formation and LVEF, LVESV, LVEDV were dynamically evaluated by GSPECT and GPET; the relationships between the size of LVA and LVEF, LVESV, LVEDV were analyzed respectively.Results: There were 5 pigs died in surgery and 2 died at the 1st week of modeling. According to golden (pathological) standard, 9 animals successfully ifnished the dynamic imaging study. At the 1st week of (basic) modeling, 4 animals formed large LVA, 2 formed small LVA at the apex and 3 without LVA formation. At the 4th and 8th weeks of modeling, dynamic imaging presented that the animals with large LVA had gradually increased range and degree of perfusion defect, LVEDV, LVESV, while gradually decreased LVEF; the above indexes were relatively stable in animals with small or none LVA. In addition, the size of LVA was related to LVEF (r=-7.26), LVEDV (r=0.855) and LVESV (r=0.825), allP<0.05. Conclusion: In experimental pigs, at the beginning of LVA formation, large range and severe perfusion defect may cause large aneurysm, the LV functional damage and remodeling may gradually increase and the prognosis is poor; in contrast, the animals with small or none LVA have better prognosis and usually without ventricular remodeling; which implies that in acute phase of LVA formation, the size of aneurysm may predict the trend of global LV systolic function and remodeling at the early stage.

AIM:To investigate the effects of fucoidan on the angiogenesis of multiple myeloma cells in vitro, and its related mechanisms .METHODS:The human multiple myeloma RPMI 8226 cells and human endothelial cells were cultured in vitro.The growth inhibition rate of RPMI 8226 cells was examined by MTT assay .The cell cycle and apoptosis rate were measured by flow cytometry .RPMI 8226 cells were treated with fucoidan for 72 h, and the cell culture superna-tant was collected .The VEGF concentration was examined by ELISA , and the tube formation assay was applied to assess the angiogenic activity .After treatment with fucoidan for 72 h at different concentrations , the protein levels of HIF-1α, VEGF, p-AKT and p-ERK1/2 were detected by Western blot .RESULTS: Fucoidan inhibited the growth of RPMI 8226 cells in a dose-and time-dependent manner .After treatment with fucoidan for 72 h, the cell cycle was arrested at G 1 phase , and the apoptotic rate of RPMI 8226 cells was increased with the increasing concentration of fucoidan , which was much higher than that in control group (P<0.05).The VEGF concentration was significantly decreased with the increa-sing concentration of fucoidan .The numbers and areas of the capillary-like structures decreased while the concentration of fucoidan increased, and those at 100 mg/L were less than those in the control (P<0.05).The protein levels of HIF-1α, VEGF, p-AKT and p-ERK1/2 in fucoidan group were significantly lower than those in control group (P<0.05).CON-CLUSION:Fucoidan inhibits the secretion of VEGF in multiple myeloma cells , and reduces angiogenesis induced by mul-tiple myeloma cells .It inhibits the protein expression of HIF-1αand VEGF , which may be related to inhibiting the phos-phorylation of AKT and ERK1/2.

Objective To investigate the effect of different gene expression levels of Syntenin on invasion and mi?gration of glioma cells and the underlying molecular mechanisms. Methods Lentiviral RNA interference was used to knockdown the expression of syntenin in U-87 cells. Real-time quantitative PCR was used to detect mRNA expression levels of syntenin . Transwell assay and adhesion assay were used to examine the invasion, migration and adhesion, re?spectively. Western-blot was used to detect the protein expression levels of Syntenin, AKT, p-AKT, and MMP-9. Re?sults The mRNA expression level of Syntenin was greatly reduced in interference group compared with empty vector group (P<0.01). The ability of invasion and migration was much lower in interference group than in empty vector group (P<0.01). However, there were no significant differences in invasion and migration between empty vector group and con?trol group. The adhesion ability of glioma U-87 cells was much higher in interference group than in empty vector group (P<0.05). However, when U-87 cells were seeded on 96-wells coated with HUVEC, the adhesion ability was much lower in interference group than in empty vector group(P<0.05). The protein expression levels of Syntenin, p-AKT, and MMP-9 in interference group were markedly decreased compared with empty vector group(P<0.05). There was no signif?icant difference in expression of AKT protein between interference group and empty vector group (P>0.05). Conclusion Syntenin may enhance the invasion and migration ability of glioma though up-regulation of p-AKT, which in turn pro?motes MMP-9 expression in a corresponding signal transduction pathway.

Objective:To explore the effect of calcium release-activated calcium channel modulator 1 (ORAI1) on the migration and invasion of colon cancer cell line SW480 and its mechanism. Methods:The SW480 cells were infected with ORAI interference lentivirus. The expression of ORAI1 mRNA and protein was confirmed by quantitative real-time polymerase chain reaction and Western blot. Transwell chamber, adhesion, angiogenesis, and vasculogenic mimicry experiments were conducted to detect the ability of cell invasion, migration, and angiogenesis and the intercellular adhesion of homogeneous and heterogeneous cells among each group. Confocal microscopy was employed to detect the difference of store-operated Ca2+entry (SOCE) in each group. Western blott was used to detect the expression of ERK1/2, p-ERK1/2, MMP-2, VEGF, and E-cadherin protein. Results:After the infection of SW480 with the ORAI1 interference lentivirus for 72 h, significant fluorescence expression was observed. Compared with the empty vector group and control group, the expression of ORAI1 was lower in the interference group (P<0.01). Invasion and migration ability decreased (P<0.01); the intercellular adhesion ability of homogeneous cells increased (P<0.05); the intercellular adhesion ability of heterogeneous cells decreased (P<0.05);the angiogenesi and vasculogenic mimicry were enhanced (P<0.01);the internal flow peak of SOCE was low (P<0.05); the expression of p-ERK1/2, MMP-2, and VEGF proteins decreased (P<0.01); and the expression of E-cadherin protein increased (P<0.01). Conclusion:ORAI1 may promote the migration and invasion of SW480. This mechanism may be associated with the increase of SOCE.

OBJECTIVETo investigate the effect of 1,25-dihydroxyvitamin D(3) and 5-fluorouracil, either alone or in combination, on the expression of IGFBP-3 in human esophageal carcinoma 109 cell xenograft in nude mice.

METHODSIn vitro cultured esophageal carcinoma Eca-109 cells were inoculated subcutaneously in BALB/c mice. The tumor-bearing mice were randomly divided into control group (A), 1,25-dihydroxyvitamin D(3) group (B), 5-fluorouracil group (C), and 1,25-dihydroxyvitamin D(3) plus 5-fluorouracil group (D). 1,25-dihydroxyvitamin D(3) and 5-fluorouracil were administered at the doses of 2.5 ug/kg and 25 mg/kg via intraperitoneal injections, respectively, and the mice in the control group received saline injection only. The tumor growth was observed and the expression of IGFBP-3 in the tumor xenograft was detected using immunohistochemistry. An automatic biochemistry analyzer was used to determine serum calcium levels, and Von Kossa staining was utilized for observation of calcium deposition in the kidneys.

RESULTSCompared with that in group A, the xenograft in groups B, C, and D all showed a lowered growth rate with a smaller tumor volume, and presented with stronger IGFBP-3 positivity and significantly higher levels of IGFBP-3 protein expression (P<0.05). In group D, the protein expression of IGFBP-3 was significantly increased compared with that in groups B and C (P<0.05). Compared with that in group A, serum calcium level was slightly increased in groups B, C, and D, , but no obvious calcium deposition was found in the kidney tissue sections.

CONCLUSIONBoth 1,25-dihydroxyvitamin D(3) and 5-fluorouracil can inhibit the growth of the tumor xenograft in nude mice, and their combination is more effective. This effect is probably associated with increased protein expression of IGFBP-3 in the xenograft tumor. No calcium deposition occurs in the kidney tissue of the tumor-bearing mice.

Animals ; Cell Line, Tumor ; Fluorouracil ; pharmacology ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Vitamin D ; analogs & derivatives ; pharmacology ; Xenograft Model Antitumor Assays

Objective To investigate the effect of integrin β4 expression on the metastasis potential of human thyroid follicular cancer cells. Methods Metastasis potential was observed and grouped in human thyroid follicular cancer cell line, CGTHW-1 cells, depending on the cells' penetrating ability in artificial matrigel. The expression of integrin β4 mRNA and protein were determined by immunocytochemistry, Western blot, and semiquantitative RT-PCR. Results The mRNA and protein expressions of integrin β4 were significantly higher in cells with high metastasis potential [0.277 5 ±0.034 0 vs 0. 187 5 ±0.022 2 ( Immunohistochemistry ), 0. 099 7 ± 0.0185 vs 0.039 0±0.010 2(Western blot), 0.555 0±0. 101 2 vs 0.270 0±0.029 9(RT-PCR), all P＜0.01]. Conclusion Integrin β4 may play an important role in human thyroid cancer invasion. It seems probably to be a potential target for human thyroid cancer treatment.

Objective:To isolate breast cancer stem/progenitor cells from human breast cancer and study their proliferation and differentiation biological characteristics over long-term passages in vitro. Methods:Human breast cancer stem/progenitor cells were enriched in suspension cultures as nonadherent mammospheres(MS). Serial sphere formation assay was performed to determine self-renewal ability of mammosphere-derived cells (MSDC). Differentiation was induced by culturing MSDC in DMEM-F12 supplemented with serum but without growth factors. The ratio of CD44~+/CD24~(-/low) cell population was evaluated by flow cytometry(FCM). Results:The mammospheres were formed after inoculation of primary breast cancer cells into culcutre medium with growth factors but without serum. The mammospheres contained undifferentiated cells similar to stem cells, which had self-renewal and extensive proliferation capabilities. With increasing passages, the cells tended to adhere and differentiate. The number of adhering and differentiating cells increased, and the amount and size of mammospheres decreased. The CD44~+/CD24~(-/low) cell population was enriched in the basal-like molecular subtype of human breast tumors. The biological behaviors of mammospheres varied between different specimens.Conclusion:Cancer cells with stem cell properties of self-renewal, indefinite proliferation and multi-lineage differentiation widely existed in human breast cancer tissues. The biological behaviors varied because of different origin of specimens and changed under the effects of environmental factors.

OBJECTIVEThe objective of this paper was to study the change of P38MAPK and Fas in the apoptosis of THP-1 cells induced by allicin.

METHODThe proliferation inhibition rates of THP-1 cells after various treatments were examined by MTT assay. Apoptosis rate was determined with Annexin V- FITC/PI double staining by flow cytometry. The expression and distribution change of the phosphorylation p38MAPK (P-p38MAPK) were detected by immunohistochemical staining. The changes of P-p38 MAPK and Fas proteins were detected by Western blot.

RESULTThe proliferations of leukemia cell line THP-1 are inhibited by allicin. MTT assay showed that allicin can inhibit the proliferation of the THP-1 cell, and the inhibition was dependent on both dose and time. The IC50 of 72 hours was 12.8 mg x L(-1). Apoptosis rate detected by Annexin V-FITC/PI was proportional to the concentration of the allicin. After the immunohistochemical staining test, the P-p38MAPK was located in the cell nucleus and plasma, showing deep brown, when adding allicin to THP-1 cell. Western blot test showed that the P-p38MAPK proteins expression was proportional to the concentration of Allicin and was also dose dependent. The levels of P-p38MAPK in negative control group, 1/2 IC50 of 72 hours group and IC50 of 72 hours group were 0.259 8 +/- 0.013 2, 0.61 2 +/- 0.008 3 and 0.505 6 +/- 0.005 5 respectively, and the levels of Fas proteins were 0.287 4 +/- 0.008 9, 0.426 8 +/- 0.007 9 and 0.597 1 +/- 0.010 9 respectively. The difference was statistically significant when compared with the negative control group (P < 0.01).

CONCLUSIONAllicin can significantly induce THP-1 cells apoptosis, and its mechanism may be related to the activation of P38MAPK/Fas.

Apoptosis ; drug effects ; Cell Line, Tumor ; Humans ; Neoplasms ; drug therapy ; metabolism ; physiopathology ; Phosphorylation ; Signal Transduction ; drug effects ; Sulfinic Acids ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

Objective To investigate the expression of platelet-derived growth factor-D (PDGF-D) and bascular endothelial growth factor(VEGF) in gastric carcinoma and elucidate the potential relationship between their overex-pression and clinical pathological characteristics. Methods Immunohistochemical assay was performed to detect the protein expression of PDGF-D and VEGF in tissues. Semiquantitative reverse transcription polymerase chain re-action (RT-PCR) was performed to evaluate the mRNA expression of PDGF-D and VEGF in part of random selec-ting gastric carcinoma samples. The correlation between the expression of PDGF-D and VEGF, and the relationship between the expression of PDGF-D, VEGF and the clinical pathological characteristics were analyzed. Results The protein expression of PDGF-D and VEGF in gastric carcinoma tissues were significantly higher than that in adjacent tissues and normal gastric mucosa(P <0.05) ;The expression of PDGF-D and VEGF correlated with TNM staging, depth of invasion and lymphatic metastasis (P < 0.05), while the expression of PDGF-D also correlated to histolog-ical differentiation (P < 0.05). There was a significant positive correlation between PDGF-D and VEGF at the mR-NA and protein expression levels (P < 0.05). Conclusion PDGF-D and VEGF specifically highly expressed in gastric carcinoma. The abnormal expression may play an important role in the carcinogenesis and metastasis of gas-tric carcinoma.

Objective To investigate the expression of platelet-derived growth factor-D(PDGF-D) and bascular endothelial growth factor(VEGF) in gastric carcinoma and elucidate the potential relationship between their overexpression and clinical pathological characteristics.Methods Immunohistochemical assay was performed to detect the protein expression of PDGF-D and VEGF in tissues.Semiquantitative reverse transcription polymerase chain reaction(RT-PCR) was performed to evaluate the mRNA expression of PDGF-D and VEGF in part of random selecting gastric carcinoma samples.The correlation between the expression of PDGF-D and VEGF,and the relationship between the expression of PDGF-D,VEGF and the clinical pathological characteristics were analyzed.Results The protein expression of PDGF-D and VEGF in gastric carcinoma tissues were significantly higher than that in adjacent tissues and normal gastric mucosa(P