OBJECTIVETo evaluate the effects and molecular mechanism of action of Changweiqing (CWQ) in reversing multidrug resistance by observing its impacts on nuclear translocation of Y-box binding protein-1 (YB-1), multi-drug resistance gene (MDR1) expression and P-glycoprotein (P-gp) expression in human colon cancer cell line HCT8/V with drug-resistance induced by vincristine.

METHODSCultured HCT8/V cells were exposed to the CWQ-containing rat serum prepared by drug perfusion. YB-1 expressions in cell plasma and nuclei were examined by Western blot; the binding activity of YB-1 to MDR1 gene promoter sequences was detected by electrophoretic mobility shift assay (EMSA); the mRNA transcription levels of MDR1, YB-1 and multi-resistance related protein (MRP) were examined by RT-PCR; the expression of P-gp on cell membrane was determined by flow cytometry. Results Along with the increasing drug's concentration of CWQ-containing serum from 1.25% up to 2.5% and 5%, the expressions of YB-1 decreased in HCT8/V cell nuclear and increased in cytoplasm gradually; the binding activity of YB-1 to MDR1 gene promoter weakened (P < 0.01), MDR1 mRNA expression and fluorescence intensity of P-gp on cell membrane attenuated (P < 0.05 or P < 0.01), while YB-1 and MRP mRNA unchanged (P > 0.05).

CONCLUSIONCWQ could reverse the drug-resistance of colon cancer cells by influencing nuclear translocation of YB-1 and reducing the expression of MDR1/P-gp.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Active Transport, Cell Nucleus ; drug effects ; Animals ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Rats ; Vincristine ; pharmacology ; Y-Box-Binding Protein 1 ; metabolism

OBJECTIVETo investigate the in vitro effects and the primary mechanisms of Changweiqing (, CWQ) on antimetastasis and antiinvasion of hypoxic colon carcinoma cells. In addition, to provide experimental evidence for the Chinese medicinal theory of "strengthening the body's resistance to eliminate pathogenic factors" in the treatment of colorectal cancer, including its invasion and metastasis.

METHODSFirst, CWQ sera were prepared with serum-pharmacology methods. Then, the modified hypoxic chamber was designed and flushed with 5% CO(2) and 95% N(2) at 37 °C to induce a hypoxic environment. The effect of CWQ serum on the viability of LoVo cells was tested with MTT cytotoxicity assay. The wound model and chamber model were established to estimate the effects of CWQ serum on migration and invasion of LoVo cells. The model for cell adhesion was established to evaluate the effect of CWQ serum on LoVo cells' adhesion. The gelatin zymography model was performed to determine the effects of CWQ serum on the activities of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). The effects of CWQ serum on the hypoxia-inducible factor 1 α (HIF-1α) nuclear translocation and the mRNA level of vascular endothelial growth factor (VEGF) in LoVo cells were determined by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses, respectively.

RESULTSCWQ inhibited LoVo cells' migration based on wound healing assay. The inhibitive effect could reach about 68.00% under hypoxic culture and about 29.87% under normoxic culture when cells were treated with 10% CWQ serum for 24 h. The results from both cell invasion and adhesion assays showed that CWQ serum could dose-dependently repress the invasion of LoVo cells and inhibit cells from adhering to extra cellular matrix (ECM). Under the hypoxic culture condition, RT-PCR analysis showed that 10% CWQ serum had down-regulated the expression of VEGF by 45.87%, and the result of Western blot analysis provided further evidence. The HIF-1α amount in the nucleus of the LoVo cells was also diminished in a dose-dependent manner, as shown by the Western blot. Gel zymogram assay revealed that CWQ serum could suppress the activities of MMP-2 and MMP-9.

CONCLUSIONSCWQ could effectively inhibit tumor metastasis in vitro The antimetastatic effects of CWQ were associated with the inhibition of cell motility, which was evidenced by inhibition of cell invasion and adhesion. The molecular mechanisms of the inhibition of tumor invasion by CWQ were due to the reduced expression of both HIF-1α and VEGF and the suppression of MMP-2 and MMP-9 expression.

Animals ; Cell Adhesion ; drug effects ; Cell Hypoxia ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Nucleus ; drug effects ; metabolism ; Colorectal Neoplasms ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Matrix ; drug effects ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Protein Transport ; drug effects ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Background and purpose:Cyclooxygenase-2 (Cyclooxygenase-2,COX-2) is an important rate-limiting enzyme that is responsible for transformation of arachidonic acid into prostaglandins(PGs).Although Helicobacter pylori(Hp) infection-induced gastric over-expression of COX-2 (COX-2) is an important factor of gastric cancer,the mechanism of COX-2 expression in gastric mucosa cells infected with Hp is still not clear.Our study was to reveal the effect of Hp on expression of COX-2(cyclooxygenase-2),the impact of p38MAPK signaling pathway in human gastric epithelial cancer cells line MKN45,and to investigate the potential mechanisms of expression of COX-2. Methods:The expression of COX-2 mRNA infection by standard Hp NCTC11637 in human gastric epithelial cancer cells line MKN45 was evaluated by real-time fluorogenic quantitative polymerase chain reaction (RFQ-PCR).The effect of infection by Hp on COX-2 expression,activation of p38MAPK and its downstream of the ATF-2 was assayed by Western blot.Results:The expression of COX-2 mRNA in MKN45 cells infected by Hp compared with control group,COX-2 mRNA had 3 fold,7.2 fold,5.1 fold,4.3 fold up-regulation after 3 hrs,6 hrs,9 hrs,12 hrs,respectively. COX-2 mRNA expression in each time group was significantly higher than that in the control group(P

Objective To analyze the correlation between SF-36 and QOL-35 and the consistency of the two kinds of life-quality questionnaires when evaluating the quality of life in patients with coronary heart disease. Methods 781 in-hospital-patients, who underwent coronary angiography from June 2008 to April 2009, were included in this study. Quality of life was measured by the Chinese versions of SF-36 and QOL-35. Demographic and clinical information were collected.Correlation Analysis was done between SF-36 and QOL-35 in the patients with coronary artery disease. Results The total score of SF-36 was 62.63 ± 12.47 and the QOL-35 total score was 62.70 ± 9.69. Data from the simple correlation analysis showed that SF-36 and QOL-35 total scores (r=0.725, P＜0.01 ), SF-36 physical functioning and QOL-35 independent living capacity (r=0.933, P＜0.01), SF-36 mental health and QOL-35 mental health (r=0.132, P＜0.01), SF-36 social function and QOL-35 social function score (r=0.215, P＜0.01 ) were all relevant. Canonical correlation analysis showed that there was significant correlation between SF-36 and QOL-35 (r=0.946, P＜0.01 ). Conclusion SF-36 and QOL-35 were consistent in the evaluation on the quality of life, at the overall level.

Objective:To observe the clinical efficacy of fire-needle therapy in treating deglutition disorders due to pseudobulbar palsy in the remission stage of stroke.Methods:Sixty-two eligible subjects were divided into a fire-needle group and a rehabilitation group by a simple randomization method at a ratio of 1:1.The two groups received same basic intervention;in addition,the fire-needle group received fire-needle treatment,while the rehabilitation group received rehabilitation training.The two groups of subjects all received a 3-week treatment and were evaluated by the dysphagia severity rating scale (DSRS),modified Mann assessment of swallow ability (MMASA) and Kubota Toshio swallow test (KTST) before and after the intervention.The complications and adverse events occurred during the trial were recorded.The data were statistically analyzed.Results:At the third week,the DSRS,MMASA and KTST scores changed significantly compared with the baseline in both groups (P＜0.05),and the changes in the fire-needle group were more significant than those in the rehabilitation group (P＜0.05).The between-group comparison at the third week showed that the therapeutic efficacy in the fire-needle group was superior to that in the rehabilitation group (P＜0.05).Conclusion:Fire-needle therapy can obviously change the DSRS,MMASA and KTST scores in pseudobulbar palsy in the remission stage of stroke,and significantly enhance the therapeutic efficacy of the treatment of deglutition disorders in this stage.

OBJECTIVETo evaluate the underlying mechanism of Jianpi Jiedu Recipe (, JJR) in the reversion of multidrug resistance concerning colorectal cancer in vitro and in vivo.

METHODSMice were treated orally with JJR at a daily 4.25 g/(kg·day) or injected with vinblastine (VCR) 2.5 mg/(kg·day) for 3 weeks after having been inoculated with HCT8/V cells; tumor tissues were assayed by hematoxylin and eosin staining. Firstly, the effects of JJR on the expression of cyclooxygenase-2 (COX-2) were tested by real-time polymerase chain reaction (PCR) technique and COX-2 gene silenced by siRNA. Secondly, the variation of intracellular concentration of oxaliplatin (L-OHP) was evaluated by the inductively coupled plasma mass spectroscopy (ICPMS) in HCT8/V and its COX-2 siRNA cells; the concentration of JJR combined with chemotherapeutic drugs and the reverse effect of multidrug resistance (MDR) in HCT8/V cells was evaluated by the MTT assay. Thirdly, real-time quantitative PCR and Western blot analysis were used to detect the multidrug resistance gene 1 (MDR1) mRNA and P-gp expression.

RESULTSJJR had an inhibitory effect on the growth of tumors in vivo, and it, in combination with chemotherapeutic drugs, could reverse the drug-resistance of HCT8/V cells and increase the sensitivity of HCT8/V cells to VCR, DDP, 5-Fu, and THP. ICP-MS results showed that JJR could increase the concentration of drugs in HCT8/V cells (P<0.01). Furthermore, it was shown that JJR could reverse drug resistance of colorectal cancer cells by decreasing MDR1 expression and P-gp level via downregulation of COX-2, which has been represented as one of the major mechanisms that contributes to the MDR phenotype (P<0.01).

CONCLUSIONJJR reversed multidrug resistance and enhanced the sensitivity to chemotherapy, which could be attributed to the down-regulation of COX-2 in MDR1/P-gp-mediated MDR colorectal cancer after chemotherapy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; drug therapy ; enzymology ; pathology ; Cyclooxygenase 2 ; genetics ; metabolism ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Green Fluorescent Proteins ; metabolism ; Humans ; Intracellular Space ; metabolism ; Mice, Inbred BALB C ; Organoplatinum Compounds ; metabolism ; RNA, Small Interfering ; metabolism ; Signal Transduction ; drug effects ; Vinblastine ; pharmacology ; therapeutic use ; Xenograft Model Antitumor Assays

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Carcinoma, Hepatocellular ; enzymology ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Liver Neoplasms ; enzymology ; pathology ; Signal Transduction ; physiology ; Time Factors

Objective To investigate the effect of different calcification patterns on the outcome of transcatheter aortic valve implantation (TAVI) by the finite element method.Methods Three calcified human aortic root models (coaptation line calcification model,attachment line calcification model and circular calcification model) were developed according to the location of calcified plaques on the aortic valve leaflets.The processes of self-expanding transcatheter aortic valve implanted into the 3 calcified models were simulated by ABAQUS software.The effects of different calcification patterns on the aortic root stresses,valve frame distortions and paravalvular gaps were analyzed.Results Circular calcification model had the largest maximum principal stress on calcified plaques (18.42 MPa),which might result in a higher risk of stroke after implantation;the circular calcification model also had the greatest distortion of the valve frame,which might lead to worse prosthetic durability;the paravalvular gaps area of the attachment line calcification model was 37.2 mm2,which was more than twice that of the other 2 models,causing more serious paravalvular regurgitation.Cenclusiens Different aortic valve calcification patterns are related to aortic root stresses,valve frame distortions and paravalvular gaps after TAVI,which will have an impact on postoperative complications and prothesis durability.The research findings provide references for the prediction of clinical outcome after TAVI.

OBJECTIVETo evaluate the accuracy of quantitative coronary angiography (QCA) assessment on target lesion and reference vessel in patients with diabetes mellitus with intravascular ultrasound (IVUS) measurements as golden standard.

METHODSQCA and IVUS were performed in 52 diabetes mellitus patients [35 males, mean age (62.3 +/- 7.1) years]. Regression equation was ascertained with the IVUS derived plaque burden as dependent and QCA derived vessel stenosis as independent variable. The measurement results derived from the two modalities on proximal and distal reference vessels were compared.

RESULTThe regression equation (constant = 0.8286, P = 0.001) of plaque burden and vessel stenosis derived from two modalities were significantly correlated (r = 0.691, P < 0.001) but QCA overestimated the stenosis severity (57.9% +/- 15.5% vs. 53.5% +/- 12.9%, P < 0.01). Target vessels negative remodeling index in these patient was 0.87 +/- 0.23. QCA significantly underestimated the proximal and distal reference segments vessel diameters [(0.81 +/- 0.24) mm, (0.64 +/- 0.17) mm, all P < 0.05] as compared to IVUS results.

CONCLUSIONDue to the significant negative vessel remodeling, QCA overestimated the stenosis severity and underestimated the reference segments vessel diameters in patients with diabetes mellitus.

Aged ; Coronary Angiography ; methods ; Coronary Artery Disease ; diagnostic imaging ; Diabetes Mellitus, Type 2 ; diagnostic imaging ; Diabetic Angiopathies ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Regression Analysis ; Ultrasonography, Interventional

OBJECTIVETo compare the value of intravascular ultrasound (IVUS) and assess the value of quantitative coronary angiography (QCA) and 64 multi-detector computed tomography (MDCT) on unstable anginas (UAP) risk stratification.

METHODA total of 61 UAP patients (low risk: 17, middle risk: 33 and high risk: 11) were recruited, 71 vessels were examined by MDCT, QCA and IVUS. Plaque characteristics (soft, fibrous, calcified and mixed plaques) and plaque burden at minimum area (< or = 50%, 51% - 74% and > or = 75%) were detected, calculated and analyzed. Results derived from various detection methods were compared.

RESULTSPlaque burden detection by QCA was comparable to IVUS results for low and middle risk UAP (r = 0.768 and r = 0.721, respectively; all P < 0.01) but not for high risk UAP (67% + or - 14% vs.75% + or - 16%, P < 0.01) due to significant positive vessel remodeling (remodeling index = 1.21 + or - 0.31). The high negative predict value of MDCT for stenosed coronary vessels (87.8% - 96.3%)was valuable for exclusion of coronary heart disease but MDCT was not able to identify fibrous cap (kappa = 0.235) and lipid core (kappa = 0.245). Extent of remodeling index, external elastic membrane area, minimum lumen area, plaque burden, plaque rupture and thrombosis increased in proportion to increasing risks of UAP patients.

CONCLUSIONSQCA is a suitable tool for assessing UAP patients with low and middle vessel stenosis but underestimated the stenosis degree in UAP patients with high vessel stenosis. MDCT is valuable for exclusion vessel disease but not useful for identifying soft and fibrous plaque. Soft plaque with positive remodeling index and minimum lumen area < 4 mm(2) derived from IVUS could correctly identify UAP patients with high degree of vessel stenosis.

Adult ; Angina, Unstable ; diagnostic imaging ; Coronary Angiography ; methods ; Coronary Vessels ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Ultrasonography, Interventional