It is possible for bone marrow cells,peripheral blood cells,myocardial cells,muscle cells and embryonic stem cells differentiating into smooth muscle progenitor cells(SPCs),which can express special marks of myosin heavy chain.SPCs remained quiescent,until mobilized in response to injury or disease.With various growth factors,cytokines,SPCs are mobilized following physiological stress and subsequently home to site of vascular damage,where they contribute to the remodeling process.Recently,various studies have found that SPCs from different sources contributed to the remodeling process of vessels and development in atherosclerosis.It is generally accepted that SPCs contribute to pathological changes,leads to restenosis.But several recent papers reported findings that injection of SPCs increased collagen and SMC content and reduced macrophage content,consistent with a more stable plaque.SPCs may secrete cytokines and growth factors both locally and systemically that enhance plaque stability.It remains to be determined whether smooth progenitor cells,which are less well studied than their endothelial counterparts,can likewise be manipulated to achieve therapeutic genes benefit.This review has elucidated the origin and distribution of SPCs,mobilization and recruitment,and their roles in cardiac vascular diseases.

AIM: To construct recombinant adenoviral vector carrying the LEDGFp52 gene by homologous recombination in bacteria and to detect its expression in vitro.METHODS: The LEDGFp52 gene was cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then, the resultant pAdTrack-CMV-LEDGFp52 was cotransfected into BJ5 183 bacteria with the adenoviral backbone plasmid pAdeasy-1. The adenoviral plasmid carrying LEDGFp52 was generated with homologous recombination in bacteria, and the adenoviruses were produced in 293 cells. These 293 cells were then infected with adenoviruses, and the expression of LEDGFp52 was detected by CPE (cytopathic effect) and western blot.RESULTS: The titer of Ad-LEDGFp52 adenoviruses was up to 5×1012 pfu/L after proliferation in 293 cells. LEDGFp52 was expressed efficiently in 293 cells after infection.CONCLUSION: The recombinant adenoviruses vector expressing LEDGFp52 was constructed successfully and can be used in further gene transfection experiments.

AIM: To construct and identify LEDGFp52 eukaryotic expression vector for RNA interference.METHODS: Recombinants were designed and established by targeting gene LEDGFp52 and plasmid pGensil-1 based on LEDGFp52 cDNA sequences of Genomes. Two pairs of oligonucleotides were synthesized according to the Tuschl principle and inserted into plasmid pGenSil-I to generate siRNA eukaryotic expression vector. DH5α strains were transformed, plasmids were extracted, and recombinant vectors were identified by the restriction map and the sequence analysis. The cultured cells were transfected by the recombinant plasmid (pGensil-1-RNA. LEDGFp52-1). At 48 hours after transfection, the whole cell protein was extracted, and the protein level was detected using Western blotting with mouse anti-human LEDGFp52 monoclonal antibody.RESULTS: Recombinant plasmids completely concord with the designs by the restriction map and the sequence analysis,the proteinlevel of LEDGFp52 was down regulated at 48hours after transfecting pGensil-1- LEDGFp52-1 expression vector into HeLa cells, the recombinant eukaryotic expression vectors were successfully constructed.CONCLUSION: siRNA recombinant can be successfully constructed by RNAi technique to inhibit the expression of LEDGFp52.

AIM: To construct the prokaryotic expression vector of rhLEDGFp52 gene,to obtain the rhLEDGF p52 protein.METHODS: rhLEDGFp52 gene was constructed into a prokaryotic expression vector pET30a (+) by recombinant DNA techniques and was identified by enzymatic digestion and sequence analysis. rhLEDGFp52 protein was induced expression by IPTG in E.coli BL21 (DE3), it was tested by Western blot and was purified by Ni-NTA His. Bind. Resin.RESULTS: We Successfully constructed the prokaryotic expression vector of rhLEDGFp52 gene and obtained its expression in E.coli BL21 (DE3),it was expressed in a soluble form and detected up to 34.63% of the total bacterial protein expressed in E.coli BL21 (DE3).Western blot analysis demonstrated that rhLEDGFp52 protein could spicifically integrate with LEDGF-ab. After purified by Ni-NTA His. Bind. Resin, the ultimate concentration of purified rhLEDGFp52 protein was 520μ g/ml and its purity was 87.93%.CONCLUSIONS: rhLEDGF p52 protein was obtained that provides an experimental basis for the further study of the biological function of rhLEDGFp52 protein.

Adenosine is an important biological substance in the body. It exists extensively in intracellular and extracellular tissues. In physiological condition, adenosine remains at very low level intissue. However, under stress such as inflammation, ischemia, hypoxia, trauma, or pain etc. the adenosine concentrationwill be elevated dramatically,indicating that adenosine participates in multiple histopathological processes. Adenosine is a natural chemical messenger that binds to four subtypes( A1, A2A, A2B, A3 ) of adenosine receptors and by that, it regulates multiple kinds of physiological functions. Studies found that adenosine plays an important role in the central nervous system, cardiovascular system and coagulation system. In recent years, adenosine has been seen as an attractive option to improve the treatment of glaucoma and retinal diseases. The effects of adenosine in ophthalmology were as follows: adjusting intraocular pressure, inhibiting retinal angiogenesis, dilating retinal blood vessels, regulating retinal nerve conduction, protecting retinal photoreceptors and ganglion cells, arresting the inflammatory response. This article discusses the research progress in adenosine and its receptors as well as biological products of adenosine and projects the application of adenosine in ophthalmology.

At the time of phage’s discovery, phage therapy was regarded as a possible treatment method against bacterial infection. Although phage therapy was used to treat and prevent bacterial infection in the former Soviet Union and Eastern Europe, it was abandoned by the West in the 1940s with the arrival of the antibiotic era. However, the ongoing evolution of bacterial multidrug-resistance has recently motivated the Western scientific community to reevaluate phage therapy for bacterial infections that are incurable by conventional chemotherapy. With the indepth study of phages, it’s increasingly acknowledged that phages, as the medicine to cure bacterial infection, are convenient, safe and efficient therapeutics. This paper summarizes the recent years’ advanced researches in this area.

Methyltransferase is an important metabolic enzyme whose main function is to catalyze the methylation of nitrogen, oxygen and sulfur atoms. It plays an important role in the metabolism of exogenous and exogenous compounds, including drugs in vivo. Methyltransferases are widely distributed in different tissues, with the liver and kidneys being the most abundant. In addition, the structure and activity of the enzyme have certain species and individual differences. This article will describe the biological properties of methyltransferases and their role in drug metabolism.

BACKGROUND:Bone marrow derived mesenchymal stem cells (BMSCs) will be homing to the lesions after balloon injury in the inflammatory reaction process.However,the molecular mechanism of transforming growth factor-β1 (TGF-β1) to promote BMSC into smooth muscle-like cell remains unclear.OBJECTIVE:To investigate the BMSC differentiation rate under different TGF-β1 levels after acute vascular injury.DESIGN,TIME AND SETTING:A randomized controlled animal experiment and in vitro induced cell observation were performed at the SPF Laboratory Animal Center and Laboratory of Tissue Engineering,Ninth People's Hospital of Shanghai Jiao Tong University Medical School between January 2008 and May 2009.MATERIALS:A total of 24 male SD rats were randomly divided into normal control and model groups with 12 animals in each group.In addition,6 4-weak-old male SD rats were used to prepare BMSCs.METHODS:Model of acute carotid artery injury was established in model group.Serum of normal control group and model group after 1,3,7 days of injury to detect TGF-β1 level after the vascular injury by ABC-ELISA.The BMSCs after one passage were cultured at a density of (1.0-3.0)×10~5 cells/100 mm in culture dish,and divided into two groups:in routine culture group,cells were cultured in high-glucose DMEM containing 20% fetal bovine serum;in TGF-β1 group,cells were induced by different concentrations of TGF-β1 (1,3,5 and 10 μg/L) based on routine culture for 1 week.MAIN OUTCOME MEASURES:Serum TGF-β1 level after the vascular injury;,cell morphological changes by inverted phase contrast microscopy;smooth muscle α-actin expression.RESULTS:The normal serum TGF-β1 level in rat was low,but increased rapidly after 1 day of injury,reached its peak at 3 days,and declined gradually but did not return to the basic level till day 7.After 1 week of induction,BMSCs were confluent,forming peak valley appearance.Immunocytochemistry showed that compared with routine culture,the rate differentiation of smooth muscle-like cell was significantly increased in cells stimulated by TGF-β1,especially 5 and 10 μg/L TGF-β1 (P ＜ 0.01).Real-time quantitative PCR results were similar to immunocytochemistry.CONCLUSION:Serum TGF-β1 level increased after acute vascular injury and peaked at day 3.In vitro,a similar concentration of TGF-β1 could induce cultured BMSC to differentiate into smooth muscle-like cells.

Objective To identify the role of autopahgy in the protective mechanism of heme oxygenase 1(HO-1) against hepatic ischemia/reperfusion (I/R) injury.Methods Forty healthy male Sprague-Dawley (SD) rats were randomly (random number table) divided into five groups (n =8 in each group),namely sham group,model group,cobalt protoporphyrin (CoPP) group,zinc protoporphyrin (ZnPP) group and 6-amino-3-methylpurine (3-MA) group.Partial hepatic I/R model was established by clamping the pedicles of left and median lobes for 1 hour and reopening for 6 hours in rats,and the rats in sham group were only received celiotomp without hepatic I/R.In the CoPP group,CoPP (a HO-1 inducer,5 mg/kg) was administered i.p 24 hours before I/R.In the ZnPP or 3-MA group,besides pretreatment with CoPP,the rats were given ZnPP (a HO-1 inhibitor,25 mg/kg) or 3-MA (an autophagy inhibitor,30 mg/kg) i.p 1 hour before I/R.Serum alanine aminotransferase (ALT) was determined with automatic biochemistry analyzer.The hepatic pathological scores (PS) were determined under light microscope using hematoxylin-eosin (HE) staining.The hepatocyte apoptosis index (AI) was assessed with terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining.Autophagosomes in liver tissue were counted under electron microscope.The mRNA expressions of HO-1,caspase-3,Beclin-1 and Atg-5 in the liver were determined by reverse transcription-polymerase chain reaction (RT-PCR).The HO-1 activity was also measured by the generation of bilirubin with the method of double-wave spectrophotometry.Results Compared with the sham group,the level of serum ALT significantly increased in the I/R group (U/L:560.3±73.6 vs.49.1 ± 13.8,P ＜ 0.01),HE staining showed a severe hepatic injury (PS:12.0±2.0 vs.1.3±0.9,P ＜ 0.01),TUNEL staining showed a higher hepatocytes apoptosis and the expression of caspase-3 significantly increased [AI:(19.38±3.07)％ vs.(3.25±1.28)％,caspase-3 mRNA (2-△△CT):4.62±0.40 vs.1.05±0.15,both P ＜ 0.01].However,there was no significant difference in the expression of HO-1 and the genes associated with autophagy between the two groups.In the CoPP group,the hepatic injury was blunted compared with that in the I/R group [ALT (U/L):223.3 ± 34.4 vs.560.3 ± 73.6,PS:5.6 ± 2.3 vs.12.0 ± 2.0,AI:(11.38± 2.39)％ vs.(19.38 ± 3.07)％,caspase-3mRNA (2-△△CT):2.42±0.33 vs.4.62±0.40,all P ＜ 0.01].HO-1 was induced in the CoPP group and autophagy was also increased significantly after I/R when compared with those in the I/R group [HO-1 mRNA (2-△△CT):3.01 ±0.71vs.1.14 ± 0.20,HO-1 activity (pmol ·mg-1 · h-1):259 ± 37 vs.113 ± 26,the number of autophagosomes:8.75 ± 0.87 vs.1.25±0.71,Beclin-1 mRNA (2-△△CT):2.85±0.28 vs.1.15±0.11,Atg-5 mRNA (2-△△CT):2.44±0.25 vs.1.14±0.12,all P ＜ 0.01].In the ZnPP group,the activity of HO-1 was much lower than that in the CoPP group,and as a result autophagy was decreased and liver injury was increased.In the 3-MA group,although there was no difference in the activity of HO-1 compared with that in the CoPP group,autophagy was inhibited,and the protective effect of CoPP was eliminated.Conclusion HO-1 could regulate the level of autophagy during liver I/R,and in turn autophagy might mediate the protective effects of HO-1 against liver I/R injury.

0.05).The recurrence of pterygium was related to the age.If the age increased five years,the risk of recurrence decreased 18.1%. Conclusion The application of MMC(during) the operation could decrease the recurrence rate of pterygium.The recurrence rate of pterygium was not related to the time of application of 0.02% MMC,and detainment for 3 min was enough during the operation.