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 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 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.

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.

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.

BACKGROUND:Neural stem cells have self-renewal and multidirectional differentiation potential, but under normal circumstances, the number of neural stem cells is less, and most cells are in the resting state. Thus, to promote the proliferation of neural stem cells is the key to the treatment of neurodegenerative diseases. OBJECTIVE:To investigate the effects of osthole on the proliferation of neural stem cells cultured in vitro, and to analyze its mechanism underlying promoting the proliferation. METHODS:Neural stem cells were cultured in vitro, and passage 3 cells were cultured with different concentrations of osthole(10, 50 and 100μmol/L). After 24 hours, cellvitality was determined by cellcounting kit-8. After 3, 5, 7 days of further culture, the radius of neurospheres was measured, and Ki67-positive cells were counted by immunofluorescence staining. Meanwhile, after 3 days of further culture, the gene expression of Notch 1, Hes 1 and Mash 1 in neural stem cells was detected by RT-PCR. RESULTS AND CONCLUSION:Compared with the control group, 50, 100μmol/L osthole could obviously promote the proliferation ability of neural stem cells. 100μmol/L osthole had the most significant effect and increased the expression of Notch 1 gene, Hes 1 gene, but it had no effect on Mash 1 gene. These results suggest that osthole can promote proliferation of neural stem cells cultured in vitro and its mechanism may be associated with activation of Notch 1 gene and Hes 1 gene in Notch signaling pathway.

OBJECTIVE:To investigate the protective effects of osthole on the nerves in model mice with craniocerebral injury. METHODS:Mice models of craniocerebral injury were established by craniotomy drill. There was a sham-operation group(isomet-ric normal saline),a model group (isometric normal saline) and osthole high,mediu,low dose groups (30,20,10 mg/kg). The drugs were given to the mice 1 h after successful establishment of the models,ip,once a day,for consecutive 14 d. Neurological severity score was conducted for the mice 12 h,3 d,7 d,14 d and 21 d after the establishment of models;HE stain was conduct-ed 7 d and 14 d thereafter and the wounds areas of brain were observed by microscope;the activity of myeloperoxidase(MPO)in the homogenate of mice’s brain tissues were determined 1 d and 3 d after the establishment of models;immunohistochemical meth-od was adopted to determine the expressions of the brain-derived neurotrophic factors (BDNF) and neurotrophic factor (NT) 3 in the mice’s brain tissues 7 d after the establishment of models. RESULTS:Compared with model group,the neurological severity scores of the mice in osthole high dose group and medium dose group were decreased 3 d,14 d and 21 d after the establishment of models;that in osthole high dose group were decreased 7 d after the establishment of models. The wounds areas of brain in osthole high dose group were smaller 7 d after the establishment of models;those in osthole high dose group and medium dose group were smaller 14 d after the establishment of models. The activity of MPO in the brain tissue in osthole high dose group was decreased 24 h and 72 h after the establishment of models.The expressions of the BDNF and NT-3 in the brain tissue homogenate in osthole high dose group and medium dose group were increased 7 d after the establishment of models,with significant differences(P<0.01 or P<0.05). CONCLUSIONS:Osthole has certain protective effects on the nerves in mice with craniocerebral injury. The mechanism may be related to improving the mice’s neurological functions,promoting wound healing,inhibiting the production of inflammato-ry factors,increasing the expression of neurotrophic factors.

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.