One slightly halophilic marine actinomycete strain JMC 06001 was isolated from a saline mud sample collected from the Island Naozhou in the South China Sea,near Zhanjiang,a city of southern China. The fermentation broth of strain JMC 06001 strongly inhibited the growth of Staphylococcus aureaus,Sarcina lutea,Bacillus subtilis,Escherichia coli,Micrococcus luteus and Proteus vulgaris.The combination of morphology,physiological and biochemical characteristics,chemotaxonomic data and 16S rRNA gene sequence analysis supported the view that strain JMC 06001 belong to a known species of the genus Steptomyces, S.peucetius.The strain studied grown well on most media tested,with white aeriel hyphae and pale yellow to pale brown substance hyphae.Yellow diffusible pigments were produced on oatmeal agar(ISP 3), potato extract agar and glucose/asparagines agar,and pale brown to deep brown diffusible pigments were produced on yeast extract/malt extract agar(ISP 2),glycerol/asparagine agar(ISP5),peptone-yeast ext-Fe agar(ISP 6)and nutrient agar.Growth occurred at 4℃～40℃and pH 6.0～9.0,with optimum growth at 28℃and pH 7.0.The tolerant range of NaCl was 0～1.5 mol/L,with best growth occurring in media containing 0.2～0.5 mol/L NaC1.

Objective To study the X-ray and CT features of costal eosinophilic granuloma for a better understanding.Methods Eight patients with costal eosinophilic granuloma proved by surgery or biopsy were analyzed retrospectively.All patients had X-ray plain film, 6 patients had CT examination,including a case of enhanced CT scan.Results All 8 lesions were solitary.Six lesions were in the anterior rib and 2 in the posterior rib.On X-ray, all case showed single cavity and oval lesion with clear boundary.On CT images, 5 lesions demonstrated expansile destruction of bone with cortical bone thinning, and 3 were osteolystic destruction with soft tissue mass around.On the patient with enhanced CT scan, the lesions showed a moderate and uniform enhancement.Conclusion The X-ray and CT findings of costal eosinophilic granuloma are characteristic.

Ultrasonic microbubbles were used to open blood-brain barriers (BBB) with a reversed and limited behavior feature in the study, which could improve the brain-targeted delivery of anti-tumor drugs. The glioma rat model was prepared. Low-frequency ultrasound was combined with microbubbles to affect the permeability of BBB compared with the permeability of independently administered Evans blue (EB) crossing BBB. Time point and length of ultrasound were investigated whether they affect the permeability of BBB and the damage of brain tissue. The effect of the growth time of glioma on BBB permeability was explored. Only glioma had a very little impact on BBB permeability. However, ultrasonic microbubbles opened the BBB with the features of temporary, limited and reversed behavior and improved EB and magnetic resonance imaging contrast agent penetrating BBB. A length of 30 s ultrasound is appropriate for opening BBB and no damage of brain tissue. Drugs should be injected before ultrasound so that they enter into brain as BBB opening. Ultrasonic microbubbles can open BBB effectively and safely, which improve drugs penetrating BBB under proper time point and length.

ObjectiveTo evaluate the safety and efficacy of tirofiba in the treatment of patients with acute ST-elevation myocardial infarction (STEMI) undergoing emergency percutaneous coronary intervention (PCI). MethodsA total of 158 patients with acute STEMI were randomly divided into tirofiban group 1 (59 cases, received tirofiban before PCI), tirofiban group 2 (56 cases, received tirofiban when PCI) and control group(43 cases, only received PCI). The coronary reperfusion flow(TIMI grade) of infarct related artery (IRA) after PCI, the resolution of the sum of ST segment elevation(sum STR) at 90 min after the procedure, the changes of myocardial enzyme at 6 h and 12 h afterwards, the left ventricular ejection fraction (LVEF) 1 week later, the major adverse cardiac events(MACE) within 30 d, bleeding and thrombocytopenia complications were analyzed and compared among the three groups. ResultsTIMI reperfusion grades in tirofiban group 1[98.3％(58/59 )]and tirofiban group 2[92.9％(52/56)]were higher than those in control group[60.5％(26/43)](P ＜0.05). The resolution of sum STR at 90 min after PCI in tirofiban group 1 [(89.3 ± 6.9)％]and tirofiban group 2[(82.4 + 7.3)％]was higher than that in control group[(65.6 +8.1 )％](P＜ 0.01 ),and there was significant difference between tirofiban group I and tirofiban group 2 (P＜0.05 ). The occurrence of MACE within 30 d was lower in tirofiban group 1 and tirofiban group 2 than that in control group (P＜ 0.05). The level of CK-MB at 6 h and 12 h afterwards was lower in tirofiban group 1 than that in tirofiban group 2,and tirofiban group 2 was lower than control group (P＜ 0.05). LVEF 1 week later in tirofiban group 1[(56.2 + 6.4)％]was higher than that in tirofiban group 2[(51.1 + 4.9)％]and control group[(49.8 + 5.7)％](P ＜ 0.05),but there was no significant difference between tirofiban group 2 and control group (P ＞ 0.05). Although bleeding incidence in tirofiban group 1 and tirofiban group 2 was higher than that in control group, no severe bleeding and thrombocytopenia was observed. Conclusion Tirofiban can safely and effectively reduce the incidence of the ischemic events in the patients with acute STEM1 during preoperative of emergency PCI.

Objective To construct the genetic recombinant adenovirus vector carrying the human growth and differentiation fac-tor-5(GDF-5) gene by using AdEasy-1 adenovirus vector system and to amplify and prepare the recombinant adenovirus in HEK 293 cells .Methods Human GDF-5 gene obtained by PCR was inserted into plasmid pMD19-T and the 1 .7 kb GDF-5 cDNA sequence was cloned into the adenoviral shuttle plasmid pShuttle-cytomegalovirus(CMV) of the AdEasy-1 system .It was identified by DNA sequencing and a digestion with Hind Ⅲ restriction enzyme .The resultant pShuttle-CMV-GDF-5 was used to generate the adenovi-ral GDF-5 vector through homologous recombination with the adenoviral backbone plasmid ,pAdEasy-1 in BJ5183 bacterial cells .It was selected by kanamycin and identified by a digestion with Hind Ⅲ restriction enzyme and amplified in XL10-Gold competent bac-teria .The DNA of recombinant adenovirus vector was finally linearized by Pac Ⅰ and the adenoviral recombinants were used to pro-duce adenoviruses in HEK293 packaging cells ,resulting in an Ad-GDF-5 identified by Western blot .The virus titer was assayed by TCID50 .Results GDF-5 cDNA sequence obtained by PCR was 1 .7 kb .Gene sequencing results indicated that the sequence was i-dentical with the one in GENBANK .Cloned sequence 1 .7 kb(GDF-5) was obtained by a digestion with Hind Ⅲ restriction enzyme after GDF-5 cDNA segment was cloned into pShuttle-CMV and AdEasy-1 .Western blot showed that two bands migrating at ap-proximately 15 and 55 kDa were clearly observed in PVDF membrane .These data confirmed that HEK293 cells expressed a large number of mature GDF-5 protein after infected with Ad-GDF-5 .Our research results demonstrated that recombinant adenovirus vector GDF-5 was successfully constructed .The virus titer was 5 .6 × 109 PFU/mL .Conclusion Recombinant adenovirus vector carrying the human GDF-5 gene is successfully constructed by using the AdEasy-1 adenovirus vector system .Moreover ,the Ad-GDF-5 with high titer is prepared .These provide the basis for further study of the biological function of GDF-5 and the gene thera-py of its related diseases .

s To compare the clinical effects of intravenously and orally administered aspirin in the treatment for acute coronary syndrome (ACS),and to evaluate the adverse effects of intravenous administration of aspirin.Methods One hundred and twenty-five patients with unstable angina pectoris or acute myocardial infarction were randomized into three groups:group 1 received intravenous aspirin (300mg/d,n =40),while groups 2 (n =42) and 3 (n =43) received orally administered aspirin (100mg/d and 300mg/ d,respectively).The control group included 30 patients with no heart disease or blood disease,and they had never taken aspirin and clopidogrel.Blood samples were taken at 2nd and 7th day of hospitalization.Platelet aggregation and the level ofplatelet activation marker CD62p were measured and compared among the groups.Patients were followed up for 6 months for the occurrence of major adverse cardiovascular events.Results There were no statistically significant differences in the decrease in adenosine diphosphate (ADP)-induced platelet aggregation rate (12.01±10.45%,6.76±14.62% and 9.73±16.72% for group 1,group2 and group 3,respectively),the decrease in arachidonic acid (AA)-induced platelet aggregation rate (6.73±11.34%,6.95±12.45% and 7.57±13.11%,respectively),and the decrease in CD62p level (10.89±18.62%,8.92±11.57% and 7.05±15.67%,respectively).At six months,there were 4 deaths (10%) in group 1,4 deaths (9.5%) in group 2 and 5 deaths (11.6%) in group 3 (P＞0.05).Conclusions Intravenous administration of aspirin provides a new approach as an anti-platelet treatment for ACS patients,especially those who can not tolerate oral administration of aspirin.(J Geriatr Cardiol 2008;5:212-216)

Objective: To evaluate the effect of acrylamide on the apoptosis of nerve cells by integral cell modelling in vitro which simulates the barrier effect and metabolic micro-environment. Methods: A non-contact and co-cultured in vitro blood brain barrier (BBB) model was established by using human umbilical vein endothelial cells (HUVEC) and rat glioma cells (C6) . The trans-endotheilal electrical resistance (TEER) and horseradish peroxidase (HRP) tracer effects were measured to verify the tight connectivity and permeability of the established BBB model. An integrate discrete multiple organ cell co-culture (idMOC) model was established by inoculating the human renal cortical proximal tubular epithelial cells (HK-2) , human normal hepatocytes (L-02) and human neuroblastoma cells (SH-SY5Y) into the self-made multi-organ plate for co-culturing. Then the model was verified by observing the growth curve of various tissue cells under co-culturing or culturing individually. SH-SY5Y cells were exposed to different concentrations of acrylamide directly and indirectly (through BBB model and idMOC model) . The changes of cell apoptosis rate were analyzed by flow cytometry to explore the impact of model on Acrylamide (ACR) injury of typical neurotoxic agents. Results: HUVEC cells can form a wide range of close-connected complex and then inhibit the external electric field under the cross-endothelial movement, and the mean was lower than that of endothelial cell culture group at 4, 5 and 6 days (P<0.05) ; After 20 min, the penetration rate of HRP in the co-culture group was less than that in the individual culture group, and the difference was statistically significant (P<0.05) , indicating that the barrier function of the co-culture group was higher than that of the individual culture group. All cells can exchange substances through the exchange hole of the culture plate, the cells grow well and there was no obvious death. The growth curve in individual culture group and co-culture group were basically the same, the difference was not statistically significant (P>0.05) . Under the condition of different concentrations of ACR (140, 270 g/ml) , compared with the direct exposure group, the apoptosis rate of the BBB model and the idMOC model were significantly decreased, and the difference was statistically significant (P<0.05) . Conclusion Based on HUVEC cells and C6 cells co-culture system, a blood-brain barrier model in vitro was established and based on co-culture of HK-2, L-02 and SH-SY5Y, the idMOC model was established. The toxicity and toxic action characteristics of ACR on SH-SY5Y cells were evaluated by validation tests.

Myofibrillogenesis regulator-1 (MR-1) is a novel protein involved in cellular proliferation, migration, inflammatory reaction and signal transduction. However, little information is available on the relationship between MR-1 expression and the progression of atherosclerosis. Here we report atheroprotective effects of silencing MR-1 in a model of Ang II-accelerated atherosclerosis, characterized by suppression focal adhesion kinase (FAK) and nuclear factor kappaB (NF-κB) signaling pathway, and atherosclerotic lesion macrophage content. In this model, administration of the siRNA-MR-1 substantially attenuated Ang II-accelerated atherosclerosis with stabilization of atherosclerotic plaques and inhibited FAK, Akt, mammalian target of rapamycin (mTOR) and NF-kB activation, which was associated with suppression of inflammatory factor and atherogenic gene expression in the artery. In vitro studies demonstrated similar changes in Ang II-treated vascular smooth muscle cells (VSMCs) and macrophages: siRNA-MR-1 inhibited the expression levels of proinflammatory factor. These studies uncover crucial proinflammatory mechanisms of Ang II and highlight actions of silencing MR-1 to inhibit Ang II signaling, which is atheroprotective.
Angiotensin II* ; Angiotensins* ; Animals ; Arteries ; Atherosclerosis* ; Cell Proliferation ; Focal Adhesion Protein-Tyrosine Kinases ; Gene Expression ; In Vitro Techniques ; Macrophages ; Mice* ; Muscle Development ; Muscle, Smooth, Vascular ; NF-kappa B ; Plaque, Atherosclerotic ; RNA, Small Interfering ; Signal Transduction ; Sirolimus

To identify the anti-bacterial compound(s) from Streptomyces hygroscopicus 17997, a geldanamycin producer, silica gel thin layer chromatography (TLC) TLC was used to separate the secondary metabolites of S. hygroscopicus 17997. Compound(s) from the silica gel TLC with anti-Gram positive bacteria activity and becoming red upon color reaction by 2.0 mol/L NaOH was analyzed by HPLC. The UV absorption profile and the retention time of a peak of HPLC were identical to those of authentic elaiophylin. A conserved region of dTDP-glucose-4,6-dehydratase (Tgd) gene was amplified by PCR from the genomic DNA of Streptomyces hygroscopicus 17997. DNA sequence analysis of the amplified DNA fragment indicated that it should be the tgd gene of elaiophylin biosynthetic gene cluster. These results implied that the compound in the peak of HPLC was elaiophylin, a macrodiolide antibiotic. The compound was then confirmed to be elaiophylin by LC-(+)-ESI-MS, which revealed that Streptomyces hygroscopicus 17997 was an elaiophylin producer. At the same time, a fast procedure, which consisted of silica gel TLC, color reaction, HPLC, PCR detection and DNA sequence analysis of tgd gene, and LC-(+)-ESI-MS, was established for rapid identification of elaiophylin and its producer.
Benzoquinones ; metabolism ; Chromatography, Liquid ; methods ; DNA, Bacterial ; genetics ; Hydro-Lyases ; genetics ; Lactams, Macrocyclic ; metabolism ; Macrolides ; analysis ; isolation & purification ; metabolism ; Mass Spectrometry ; methods ; Sequence Analysis, DNA ; Streptomyces ; genetics ; isolation & purification ; metabolism