Aim To investigate the reversal effect of vatalanib, a novel kinase inhibitor, on multidrug re-sistance in cancer cells and its mechanism. Methods The cytotoxicity and reversal effects of vatalanib were evaluated in both resistant and sensitive tumor cell lines by MTS or SRB assays. The intracellular accumu-lation of fluorescence substrates ( Rh-123 , MX and ADR for P-gp, BCRP, MRP1, respectively) were ana-lysed by flow cytometry. Western blot or qRT-PCR was
used to determine the protein or mRNA expression lev-el of BCRP. The effect of vatalanib on ATPase activity of BCRP was determined using crude membranes pre-pared from HEK293/ABCG2 cells. Results Vata-lanib at the nontoxic dose ( 5 μmol · L-1 ) potentially reversed BCRP-mediated MDR in cancer cells, howev-er it had no effect on P-gp or MRP1 mediated MDR. Vatalanib did not alter the intracellular accumulation of MX in HEK 2 9 3 / ABCG 2 , and had no influence on the
BCRP-mediated drug efflux. The ATPase assay indica-ted that vatalanib may serve as a substrate of BCRP. Furthermore, vatalanib dramatically suppressed levels of both the protein and mRNA expression of BCRP in concentration-and time-dependent manners. However, reversal concentration of vatalanib had no influence on the total and phosphorylated forms of AKT and ERK1/
2 in resistant cancer cells. Conclusion Vatalanib could significantly reverse BCRP-mediated MDR with specificity, and its mechanism may correlate with the down-regulation levels of BCRP both mRNA and pro-tein in resistant cancer cells.

Objective To observe any effect of low intensity pulsed ultrasound (LIPUS) on the expression of integrin-focal adhesion kinase (FAK)-mitogen-activated protein kinase (MAPK) mechanochemical transduction pathway-related proteins in the chondrocytes of rabbits with knee osteoarthritis (OA).Methods Of the 18 New Zealand white rabbits selected for the study,twelve received knee anterior cruciate ligament transection to model OA.The remaining 6 rabbits served as normal controls.At the 4th week after modeling the rabbits were sacrificed and chondrocytes were isolated and cultured in vitro.All the cultured cells were randomly divided into three groups:a normal control group (NC),an OA model group (OA) and an OA model plus LIPUS group (OA + LIPUS).When the cells had been cultured to the 2nd passage,the NC group and OA group cells had received no treatment.The OA + LIPUS group cells were exposed to 40 mW/cm2 of LIPUS for 20 min,once a day for 6 days.The expression of collagen protein type Ⅱ,aggrecan,MMP-13,integrin β1 p-FAK and p-p38,p-ERK1/2 and p-JNK Mapks were detected by Western blotting.Results Compared with the NC group,the expression of collagen type Ⅱ and aggrecan was significantly lower in the OA and OA + LIPUS groups,with more significantly lower expression of collagen type Ⅱ and aggrecan in the OA group than in the OA + LIPUS group.Compared with the NC group,the expression of MMP-13 was significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA group.Compared with the NC group,the expression of integrin β1 and p-FAK was also significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA + LIPUS group.Compared with the NC group,the expression of p-p38,p-ERK1/2 and p-JNK was also significantly higher in the OA group,but compared with the OA group,the expression of those kinases was,on average,significantly lower in the OA + LIPUS group.Conclusions LIPUS can inhibit the degradation of collagen type Ⅱ and aggrecan,and inhibit the expression of MMP-13,p-p38,p-ERK1/2 and p-JNK in OA chondrocytes,at least in vitro.At the same time,LIPUS can increase the expression of integrin β1 and p-FAK.The results show that LIPUS may activate an integrin-FAK-MAPK mechanochemical transduction pathway to induce changes in the extracellular matrix of chondrocytes.

Objective To observe the effect of low intensity pulsed ultrasound (LIPUS) on chondrocytes co-cultured with infrapatellar fat pads.Methods Twenty-four infrapatellar fat pads and cartilages from female knee trauma patients aged between 25 and 35 were cut and stained using hematoxylin-eosin staining.Chondrocytes were isolated from part of the integrated surface of the cartilages to be cultured in vitro.They were then randomly divided into a normal chondrocyte group (the control group),a normal chondrocyte+FCM (fat conditioned medium) group (the model group),a normal chondrocyte+ FCM + LIP US group (the treatment group),and a normal chondrocyte+ FCM + LIPUS + GRGDSP (an integrin inhibitor) group (the inhibited group).The treatment group and inhibited group received LIPUS at 40 mW/cm2 for 20 min once a day,while the other groups received sham LIPUS treatment.Five days later,the cells were collected and western blotting was used to examine the expression of type Ⅱ collagen (COL2),aggrecan (Acan),matrixmetalloproteinase (MMP)-13,integrin β1,phosphorylation-focal adhesion kinase (p-FAK) and phosphorylation p38 (p-p38).Results Western blotting showed that compared with the control group,the expression of COL2 and Acan was significantly lower in the model group,but that of MMP-13,integrin β1,p-FAK and p-p38 was significantly higher.As compared with the model group,the expression of COL2,Acan,integrin β1 and p-FAK was significantly higher,and that of MMP-13 and p-p38 was significantly lower in the treatment group.The expression of COL2,Acan,MMP-13,integrin β1,p-FAK and p-p38 showed no significant difference between the inhibited and model groups,but that of COL2,Acan,MMP-13,integrin β1,p-FAK and p-p38 was significantly different between the control and treatment groups.Conclusions LIPUS provides a protective effect on chondrocytes through inhibiting the expression of MMP-13 induced by adipokines and the degradation of COL2 and Acan through activating the integrin-FAK-p38 MAPK pathway.

Objective To observe the effects of modifiedDanshen Decoction on spermidine/spermine acetyltransferase (SSAT) /polyamine pathways of SD rats with IRI; To investigate its protective mechanism. Methods The model of IRI was established by ligating left anterior descending coronary artery for 30 min followed by reperfusion for 90 min. The SD rats were randomly divided into the control group, sham-operation group, model group and modifiedDanshen Decoction group, with 10 rats in each group. The myocardial infarction size was measured by using TTC staining. The contents of SSAT were measured by ELISA. The SSAT mRNA and SSAT protein expression level were detected with real-time fluorescent quantitative PCR method and Western blot, respectively. The contents of polyamines (putrescine, spermidine, spermine) in cardiac tissue were detected by HPLC. Results Compared with sham-operation group, the myocardial infarction size, the SSAT content, the SSAT mRNA and SSAT protein expression levels of model group increased significantly, the contents of polyamines decreased significantly, with statistical significance (P<0.01); Compared with model group, the myocardial infarction size of modifiedDanshen Decoction group was significantly reduced, while the SSAT content and SSAT mRNA and protein expression level decreased significantly, the contents of polyamines increased, with statistical significance (P<0.05, P<0.01).ConclusionModifiedDanshen Decoction can adjust the SSAT polyamine pathways and increase polyamine content in cardiomyocytes, and thus play a role of protection of myocardial ischemia-reperfusion injury.

Alleles ; Animals ; Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Cytochrome P-450 CYP2A6 ; Humans ; Mixed Function Oxygenases ; genetics ; metabolism ; Nicotine ; metabolism ; Polymorphism, Genetic ; Tobacco Use Disorder ; metabolism

According to the patient examination criterion and the demands of all related departments, the DSA digital subtraction workstation has been successfully designed and is introduced in this paper by analyzing the characteristic of video source of DSA which was manufactured by GE Company and has no DICOM standard interface. The workstation includes images-capturing gateway and post-processing software. With the developed workstation, all images from this early DSA equipment are transformed into DICOM format and then are shared in different machines.
Angiography, Digital Subtraction ; instrumentation ; methods ; Image Processing, Computer-Assisted ; Software Design ; User-Computer Interface

AIMTo observe the influence of polysaccharides of Angelica sinensis (ASP) on the immunologic function of rat Kupffer cells.

METHODSNormal rat Kupffer cells were treated with ASP in vitro. Absorbance at 540 nm ( A540) of neutral red absorption and supernatant NO, TNF-alpha in the cells were measured to evaluate the immunologic function of Kupffer cells; LDH leakage was measured to estimate the severity of cellular damage; Rats were given ASP 0.025, 0.1, 0.25 and 1.0 g x kg(-1) ig (qd x 7 d) in vivo. The above indices and ACP of Kupffer cells were measured, sGST and sALT activity were detected as indices of hepatotoxicity.

RESULTSASP markedly enhanced the phagocytic activity, ACP and supernatant NO, TNF-alpha of Kupffer cells both in vitro and in vivo . The increase of sGST was observed after administration of ASP 1.0 g x kg(-1), but the LDH leakage of the hepatocytes was not increased in vitro.

CONCLUSIONASP with suitable dose could activate the function of Kupffer cells. Slight liver injury was caused by ASP 1.0 g x kg(-1) in vivo, which was likely caused by factors, such as NO, TNF-alpha, indirectly.

Adjuvants, Immunologic ; pharmacology ; Alanine Transaminase ; blood ; Angelica sinensis ; chemistry ; Animals ; Cells, Cultured ; Female ; Glutathione Transferase ; blood ; Hepatocytes ; metabolism ; Kupffer Cells ; immunology ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Nitric Oxide ; metabolism ; Phagocytosis ; drug effects ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; metabolism

Objective To investigate the various factors of clinical competence model of famous and veteran physicians of Chinese medicine, and to mine effectively the clinical experience of modern famous and veteran physicians of Chinese medicine.Methods The clinical experiences of famous and veteran physicians of Chinese medicine were determined by applying the methods of philology, psychology and statistics. The clinical competence model of famous and veteran physicians of Chinese medicine was established and its reliability and validity were tested with the expert experience evaluation.Results The codes of various typical behaviors in 42 medical records showed a high consistency. There were six competence factors confirmed according to the typical behaviors. The clinical competence model of famous and veteran physicians of Chinese medicine was established. The test result indicated that the model had a higher validity.Conclusion A complete clinical competence model of famous and veteran physicians of Chinese medicine contains six competence factors, including the abilities of clinical communication, clinical information collection, clinical thinking, patient orientation, diagnosis and treatment, and research and innovation. The model also includes standard operational definition of every competence factor, and clinical typical behaviors and explanation reflecting the competence factors.

Aim:To investigate the effect of Hemangiopoietin (HAPO) on the hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice.Methods: Balb/c mice were underwent total body irradiation at 700 cGy 137Cs ? radiation and were treated with HAPO or recombinant human granulocyte colony stimulating factor (rhG-CSF) after irradiation. The hematopoiesis reconstitution of mice were detected. Cells from bone marrow of Balb/c mice were cultured with HAPO or rhG-CSF for 24 hours or 72 hours before or after the cells were irradiated. The viability of cells were assessed and the ability of in vitro hematopoiesis reconstitution were also detected. Result: rhG-CSF and HAPO treated mice both showed increased survival rate and increased colony forming units. The peripheral WBC number increased greatly. The HAPO group was most quickest compared with rhG-CSF group and PBS control group. The number of bone marrow cells at day 14 of rhG-CSF group was higher than that in HAPO group, but the number of bone marrow cells at day 32 of rhG-CSF was lower than that in HAPO group. The number of bone marrow cells at day 42 of rhG-CSF was below normal. The number of bone marrow cells at day 42 of HAPO group was nearly normal. The number of CFU-GEMM in HAPO group was most compared with that in rhG-CSF group and PBS control group at day 7, 14 and 21 after radiation. The survival rate of cells after radiation in HAPO group was markedly higher than that in PBS control group, but the survival rate of cells after radiation in rhG-CSF group was no notable difference compared with that in PBS control group. In MTT assay, both HAPO and rhG-CSF incubation stimulated proliferation of bone marrow cell at 72 hours after radiation. Bone marrow cells formed Hematopoietic islands in HAPO group after radiation and were positive for sca-1 and CD31. CD31 positive endothelial cells increased around the Hematopoietic islands. There was no Hematopoietic islands formation, few CD31 positive endothelial cells and no sca-1 positive cells in PBS control group. Conclusion: HAPO can promote hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice. It can increase the survival rate of mice and stimulate the proliferation of hematopoietic stem cells.

Objective To screen the effective parts of loosening bowel and relieving constipation in mice from different extracted parts of Angelica and to investigate the utilization of fuzzy matter-element model in evaluating the effect of moisturizing intestine to relaxing the bowels of Angelica's different extracted parts.Methods Kunming mice were randomly divided into 11 groups,including normal group,model group (constipation),positive control group (Nongsuo Danggui Wan and Runchang Wan:20 μL · g-1),medicated group consisted by 7 different extracted parts of Angelica (3.4 g · kg-1 Angelica herbs).The constipation model was formed by the compound difenoxate solution 5.0 mg · kg-1 and finished after 30 min.We used the first defecation time as well as the dark stool points in 6 h,the weight of stools,the pushing rate of intestine and intestinal moisture as detection index and then using fuzzy matter-element model on variation coefficient weight to evaluate the comprehensive effect of different extract parts of Angelica.Results Time of first black stool discharge as well as the dark stool points in 6 h,the weight of stools,the pushing rate of intestine and intestinal moisture in model group were (241.90 ± 48.10) min,(9.00 ± 5.40) count,(149.10 ± 86.80) mg,(48.90 ± 16.50) ％,(1.45 ± 0.16) g,respectively.The 70％ ethanol impregnation part (part 2) on the factors were (172.60 ± 40.70) min,(17.30 ± 4.50) count,(223.70 ± 63.40) mg,(73.70 ± 13.10) ％,(1.71 ± 0.18) g,respectively.The difference between part 2 and model group was statistically significant in the factors (P ＜0.05 or P ＜0.01).Combining with the result of the value of Euclid approach degree,the maximum is 0.71 (part 2:70％ alcohol impregnation part).Conclusion The 70％ alcohol impregnation part is the effective part of loosening bowel to relieve constipation in mice.The method and model can be used to objectively evaluate the effect of moisturizing intestine to relaxing the bowels of Angelica' s different extracted parts.