Objective To identify the causes of coronary angiographic hazy lesions by intravascular ultrasound (IVUS) to avoid inappropriate stenting. Methods Twenty-five cases with hazy regions on coronary angiogram were consecutively identified from July 2009 to March 2010. Hazy regions were defined by coronary arteriongraphy as reduced contrast density without a clearly defined intimal tear, dissection,thrombus,or stenosis ( ＞ 50% ). This cohort of patients were subsequently underwent IVUS examinations and treated according to the results of IVUS. Results The lumen CSAs were settled as ＞ 4. 0 mm2 in all examinations. Among all 25 cases,hazy lesions were located in left anterior descending in 12 patients, right coronary artery in 6 patients, left circumflex in 5 patients, and left main artery in 2 patients. According to the IVUS findings, 2 cases showed absolutely normal or near-normal arterial wall structure image, 10 cases showed calcified plaque,5 cases showed plaque rupture,3 cases showed eccentric plaque ,2 cases showed thrombosis formation,2cases showed dissection,1 case showed subintimal hematoma. Seven patients received stent implantation, and the rest accepted medical therapy. There were no in-hospital MACEs reported among all patients. Conclusion Nearly half of the coronary arteriongraphic hazy lesions were caused by calcified plaque. IVUS can distinguish calcified plaques from intimal tears, thrombus and other underlying etiologies,and help to avoid unnecessary stenting.

Objective To observe the effectiveness and safety of clopidogrel combined with fibrinolytic therapy for acute ST segment elevation myocardial infarction(STEMI).Methods From January 2004 to December 2006,a total of 158 STEMI patients in our hospital were treated with fibrinolytic agents combined with or without clopidogrel.There were 84 patients in clopidogrel group,including 66(78.6%)male,18(21.4%)female and 45(53.6%)elderly patients(aged ≥ 60 years);74 patients were in control group,including 58(78.4%)male,16 female(21.6%)and 40(54.1%)elderly patients.Clinical characteristics,patency of infarct-related artery(IRA),30 d major adverse cardiac events(MACE),recurrent angina and hemorrhage events of the two groups were retrospectively analyzed.Efficacy and safety of fibrinolytic therapy combined with clopidogrel in elderly patients(≥60 years old)subgroup were also estimated.Results Baseline clinical characteristics between the two groups were comparable.Clopidogrel therapy was associated with a higher proportion of patent IRA(69.1% vs 51.4%,P=0.035)and lower rates of cardiac death(0 vs 6.8%,P=0.047)and MACE(2.4% vs 12.2%,P=0.036).There was no significant difference in the incidence of recurrent angina(3.6% vs 2.7%,P=0.885),slight and severe hemorrhage events(7.1% vs 9.5%,P=0.811;1.2% vs 1.4%,P=0.533)between the two groups.In elderly patients,clopidogrel therapy was associated with a higher proportion of patent IRA(68.9% vs 45.0%,P=0.045),the declining tendency in MACE(2.2% vs 12.5%,P=0.155),but no significant differences in the other indexes.There were no significant differences in above-mentioned indexes between the elderly and younger patients in clopidogrel group.Conclusion Addition of clopidogrel to fibrinolytic therapy is safe and effective for both the elderly and younger STEMI patients.

Tumor blood vessels are characterized with insufficiency and distortion. Endothelial molecules of neovascularization and the heterogeneity of microvasculature in tumor interstitial are the basis for molecular-targeted ultrasonic evaluation of tumor angiogenesis, and has become focus in recent years. Research progresses of molecular-targeted ultrasonic imaging in angiogenesis and heterogeneity of tumors were reviewed in this article.

Objective To investigate the effects of osthole on neural stem cells ( NSCs) differentiation and explore the potential mechanism. Methods Brain-derived NSCs from newborn mice were isolated and cultured in vitro and determined by immunofluorescence. The P5 generations of NSCs were placed in culture solution with osthole at concentrations of (0,10,50, 100 μmol·L-1 ) . The neuron, astrocyte and oligodendroglia cell differentiation were determined by immunofluorescence. The mRNA expression of Notch 1 and its target genes Mash 1 and Neurogenin 2 were assessed by RT-PCR. Results The neurosphere displayed Nestin and Sox 2-postive by immunofluorescence, suggesting that the cultured cells were NSCs. Osthole promoted NSCs differentiating into more neuron(P<0. 01) and oligodendrocyte(P<0. 05), but not astrocyte. Meanwhile, osthole significantly reduced the mRNA expression of Notch 1(P<0. 01) and increased Ngn 2(P<0. 01)at the dose of 100 μmol·L-1. Conclusion Osthole enhances NSCs differentiating into more neuron and oligodendrocyte via probablly inhibiting Notch signal pathway.

Aim To investigate the effects of neurotro-phin-3 ( NT-3 ) gene overexpression on the differentia-tion into cholinergic neuron of neural stem cells ( NSCs) in vitro and its underlying mechanism. Meth-ods Brain-derived NSCs from newborn mice were iso-lated and cultured in vitro and determined by immuno-fluorescence. The NSCs were divided into three groups: NSCs, GFP-NSCs and NT-3-NSCs groups. The expression of NT-3 was detected by immunofluo-rescence and ELISA. Then, the ability of NSCs on dif-ferentiation into cholinergic neuron was detected by im-munofluorescence and RT-PCR, and the Acetylcholine Assay Kit was used for acetylcholine ( ACh) , and the expression of Hes1 , Mash1 and Ngn1 mRNA was de-termined by RT-PCR. Results The neurosphere dis-played Nestin and Sox 2-postive by immunofluores-cence, suggesting that the cultured cells were NSCs. The proportion of ChAT immunopositive cells was sig-nificantly higher in the NT-3-NSCs group than that in the other two groups ( P <0. 01 ) . Ach secretion in NT-3-NSCs was significantly elevated compared with the other two groups ( P <0. 01 ) . NSCs transfected with NT-3 increased the levels of Mash1 and Ngn1 mR-NA, and decreased the level of Hes1 mRNA ( P <0. 05 ) . Conclusion NT-3 can significantly promote the in vitro differentiation of NSCs into cholinergic neu-rons via probablly inhibiting Notch signaling pathway.

AIM:To explore the protective effect of osthole on the SH-SY5Y cells transfected with APP595/596 gene, and to investigate the molecular mechanism.METHODS:The SH-SY5Y cells were transfected with APP595/596 gene in vitro for establishing a cell model to study the pathogenic role of amyloid β-protein ( Aβ) .The cell viability was detected by CCK-8 assay.The release of lactate dehydrogenase ( LDH) was determined by the colour reaction of dia-phorase-INT.The cell apoptotic rate was analyzed by flow cytometry.The expression of β-site APP cleaving enzyme 1 ( BACE1) at mRNA and protein levels was detected by RT-PCR and Western blot.The expression of Aβwas measured by the technique of immunofluorescence cytochemistry and Western blot.RESULTS: Treatment with osthole inhibited the LDH release, and increased the viability of the cells.The percentage of apoptotic cells was also significantly decreased. Osthole also inhibited the expression of BACE1 at mRNA and protein levels and the protein expression of Aβ.CONCLU-SION:Osthole has protective effect on SH-SY5Y cells transfected with APP595/596 gene.The mechanism may be associ-ation with inhibiting the mRNA and protein expression of BACE1.

Aim To investigate the effects of osthole ( Ost) on the ability of proliferation and differentiation in APP transduced neural stem cells( NSCs) , and neu-ronal apoptosis, in order to find related mechanism. Methods A model of Alzheimer′s disease( AD) cells was successfully established by transducing APP gene into NSCs in vitro. The ability of proliferation and dif-ferentiation was tested by staining. The viability of NSCs was determined by using CCK-8 assay. The cell apoptosis was tested by Hoechst 33258 staining. The expression of GSK-3β and β-catenin mRNA was deter-mined by RT-PCR. The expression of GSK-3β and β-catenin protein was determined by Western blot. Re-sults The ability of proliferation had increased by 10 . 24% with Ost treatment, compared with APP group. The ability of differentiation had increased by 6 . 74%with Ost treatment, compared with APP group. The vi-ability of NSCs had increased and cell apoptotic rate had decreased significantly. From the results of RT-PCR and Western blot, we could find the expression of GSK-3βmRNA and protein had decreased, and the ex-pression of β-catenin mRNA and protein had increased significantly, compared with APP group. Conclusion Ost could enhance the ability of proliferation and dif-ferentiation into more neurons of NSCs transducing APP gene, and reduce neuronal apoptosis. It might be relat-ed with activiting Wnt/β-catenin signaling pathway.

Objective To screening optimized ultrasound (US)condition for MCF-7 cells and MCF-7/Tax cells,and to investigate the effect of dual-ligands nanoparticles (NPs ) combined with US on overcoming multidrug resistance (MDR).Methods To obtain optimized US condition,cell viability and cellular uptake of NPs treated with MCF-7 cells and MCF-7/Tax cells were detected under different ultrasonic parameters including ultrasound intensity,exposure duration and microbubble concentration. MTT assay was performed to observe the effect of NPs combined with US on cells.Results The optimal US conditions for MCF-7 cells was irradiation intensity 0.6 W/cm2 ,irradiation time 80 s,and the MBs concentration with volume ratio of microbubbles to medium was 1∶1 0;and for MCF-7/Tax cells,irradiation intensity 0.6 W/cm2 ,irradiation time 80 s,and the MBs concentration with volume ratio of microbubbles to medium was 1∶20.Compared with individual NPs,NPs assisted by US exhibited greater killing ability for MCF-7 (IC50 1 .43 ng/ml)and MCF-7/Tax cells (IC50<800 ng/ml).Conclusions US effect could enhance drug sensitivity of the dual-functionized NPs for MCF-7/Tax cells,indicating that US irradiation had potential assisted modality to reverse tumor MDR.

Objective To prepare a new kind of targeted liposome ultrasound contrast agent with small peptide K237 as the ligand which can combine specifically with KDR which is the main receptor of VEGF.and to test its capability in vitro. Methods Targeted bubbles(P-Bio-Av-Bio-Mbs) were formed through "biotin-avidin" bridge grafting, then they were incubated respectively with LOVO, HUVECs and LS174T which were KDR positive or negative expressed in various cells,meanwhile incubated LOVO cells with FITC- P-Bio-Av-Bio-Mbs,FITC-P-Mbs and FITC-Mbs respectively. After that, the rosette formation rate and fluorescence intensity of the combination between microbubbles and cells were observed with microscope and fluorescence microscope. After being incubated with small peptide K237 of 10 μg and 50 μg, LOVO cells were incubated with P-Bio-Av-Bio-Mbs for observing the distribution of microbubbles. Results In KDR sharply positive expressed LOVO cells, the surrounding rosette formation rate was as high as 90. 52% with the fluorescence intensity of grade 3, and it was 53. 46% with grade 2 fluorescence intensity rate in KDR positive expressed HUVECs cells, while in KDR negative expressed LS174T cells, there were few microbubbles surrounded with rosette formation rate of 5. 57% and fluorescence intensity rate of grade 0-1, therefore there were significant statistic differences in rosette formation rate among groups ( P < 0.05). After LOVO cells combined with FITC-P-Bio-Av-Bio-Mbs, FITC-P-Mbs and FITC-Mbs respectively,there were significant differences in their rosette formation rate, namely 89.62%, 7. 56% , 0 with the fluorescence rate of 3,0 - 1 and 0 respectively. Targeted cells pretreated with 10 pg K237 showed significant decreased rosette formation,and there was no formation in 50 ?g pretreated group. Conclusions KDR-Targeted liposome contrast agent with small peptide K237 liganded has been successfully prepared through biotin-avidin mediation and could combine specifically and high efficiently with targeted cells in vitro. The KDR-targeted molecular imaging of tumor neovascularizaiton may provide a new approach for early diagnosis of carcinoma.

Aim To investigate the effects of osthol on cell apoptosis and inflammatory cell infiltration after brain stab wound injury in mice. Methods The mice underwent the stab wound injury by a needle, then were randomly divided into sham operation group, model group, osthol 10, 20, 30 mg · kg-1 treatment group. The main examinations included mice brain wa-ter content; the apoptotic cytokines Bax, Bcl-2, Caspase-3 mRNA expression were assessed by PT-PCR; immunohistochemistry staining was used to de-tect neutrophils (MPO) and microglia (Iba-1) infiltra-tion and Caspase-3 positive cell expression around in-jured lesions. Results Treatment with osthole 20, 30 mg·kg-1 group significantly reduced the water content in injured brain, improved the ratio of Bax/Bcl-2, and reduced the expression of apoptosis cytokine Caspase-3 mRNA. Osthole 30 mg·kg-1 treatment group obvious-ly reduced the infiltration of neutrophils and microglial cells and significantly reduced the number of apoptotic cells around the injured cerebral cortex. Conclusion Osthole has therapeutic effect on stab wound injury in mice, and the possible mechanism may be by reducing the infiltration of inflammatory cells and reducing apop-totic cells.