Objective To construct an expression vector harboring CYP2B1 suicide gene, and detect its expressions in tumor cell lines. Methods PCR amplification was performed using primers based on murine CYP2B1 gene sequence from gene bank and pc3/2B1 as template. PCR product was directly inserted an eukaryotic expression plasmid pcDNA3.0. The recombinants were analyzed and identified by restriction enzyme analysis, PCR and sequencing. Then the recombinant vector pcDNA3.0/CYP2B1 was transfected into three tumor cell lines by liposome-mediated method. The expressions of CYP2B1 gene in all the cell lines were detected by RT-PCR method. Results pCDNA3.0/CYP2B1 vector was successfully constructed, and could express CYP2B1 mRNA in the three tumor cell lines. Conclusion Eukaryotic expression vector pcDNA3.0/CYP2B1 containing CYP2B1 gene under the control of a CMV promoter is an novel effective expression vector for tumor gene therapy.

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.

AIM:To explore the mechanism by which over-expression of enhancer of zeste homolog 2 (EZH2) in a panel of gastric cancer cell lines is involved in tumorigenesis of gastric cancer .METHODS: Real-time PCR and Western blot were employed to examine the mRNA and protein levels of EZH 2, respectively.MTS assay, cell migration and soft agar assay were performed to investigate the role of EZH 2 in the regulation of stomach cancer behaviors .The effect of EZH2 on NF-κB target gene expression was determined by Luciferase reporter and real-time PCR.Co-immunoprecipitati-on was used to analyze the interaction of EZH 2 and p65 in HEK293T cells.RESULTS: The expression levels of EZH2 were significantly increased in the gastric cancer cells compared with normal gastric epithelial cells .Pharmacological inhibi-tion by DZNep or knockdown of EZH2 significantly compromised AGS and SNU-16 cell activity , cell migration and anchor-age-independent cell growth.Moreover, siRNA knockdown of EZH2 impaired NF-κB downstream targets, such as IL-8, CXCL5 and CCL20.In addition, the interaction of EZH2 and p65 was detected.CONCLUSION: EZH2 mediates the growth of gastric cancer cells through the regulation of NF-κB downstream gene expression .

BACKGROUND: Foreign studies have demonstrated that the blue light at 470 nm inhibits melatonin secretion and displays the most obvious biorhythm regulation. To date, light-emitting diode (LED) applied in regulating biorhythm remains poorly explored. OBJECTIVE: To explore whether a certain intensity of LED (blue) light could induce retinal injury in rats. DESIGN, TIME AND SETTING: Randomized, controlled animal experiment was performed at the Animal Laboratory of Peking University Third Hospital between May 2007 and April 2008. MATERIALS: A total of 32 SD rats and 16 BN rats were provided by Animal Department of Peking University Third Hospital. METHODS: A total of 16 SD and 16 BN rats were respectively randomly divided into test and control groups. Test group rats were placed in light boxes which were controlled by blue LED (wavelength 470 nm) at a intensity of 300-350μW/cm2, 4 hours everyday for 3 days. The remaining SD rats were placed in light box which was controlled by blue LED (wavelength 470 nm) at a intensity of 120-150μW/cm2, 4 hours everyday for 3 days. The control rats were not treated. MAIN OUTCOME MEASURES: At the second day after light irradiation, the rats of all groups were sacrificed and both eyeballs were harvested. The frozen sections were subjected to hematoxylin-eosin staining to observe changes of rat retina. RESULTS: A total of 48 rats were included in final analysis. The retina of SD rats became thinning and disorderly arranged following blue LED irradiation at density of 300-350μW/cm2, but the retina of BN rat remained unchanged similar to control group. After blue LED irradiation at density of 120-150μW/cm2, the retina of SD rat remained unchanged similar to control group. CONCLUSION: Blue LED light source irradiation at a intensity of 300-350μW/cm2 is safe to pigment-protected retina, and at a intensity of 120-150μW/cm2 does not injury retina of different races of rats.

This paper presents a description of the rules of construction and function of genome as well as evolutional relationship between organisms, and opens out the essence of life through introducing the progress in genome of model organism. The other purpose of this review is to highlight the status and function of model organism in the research of comparing genomics so as to provide the model of cycle for researches into high creature life, especially human beings life.
Animals ; Chromosome Mapping ; Genes ; genetics ; Genome ; Genomics ; methods ; trends ; Human Genome Project ; Humans ; Models, Animal

Objective To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library. Methods Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT)15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis. Results The amino acid sequence, encoded by the positive clone with a poly (A) 22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B. napus, fission yeast, A. thaliana, C. elegans and fruit fly up to 71.6%. Conclusion The protein, RNA polymerase subunit encoded putatively by the clone, is high conservative in different species.

Objective To evaluate the clinical value and safety of angiography and embolization in the treatment of pulmonary sequestration causing massive haemoptysis. Methods Though digital subtraction angiography(DSA), abnormal arteries were demonstrated in 12 cases with massive haemoptysis. All the abnormal arteries were embolized by gelatin sponge plus silk thread sterilized with high temperature and pressure. Results Among 12 cases of the pulmonary sequestration, 26 abnormal arteries were discovered, originating from thoracic aorta and presenting enlargement, twisting and irregular diameter with strands of middle and distal arterial segments associated with abundant vasculature network. There were several arteries supplying the lesion in 11 cases, and massive haemoptysis were stopped after embolization. Follow up for 6 to 18 months, no recurrence and no complication occurred. Conclusion Arterial angiography and embolization with gelatin sponge plus silk thread for treating pulmonary sequestration with massive hemoptysis possesses high clinical efficiency and safety.

Objective:To identify the key genes related to rheumatoid arthritis (RA) by to the weighted gene co-expression network analysis (WGCNA) and experimental verification to find key genes related to RA.Methods:The microarray data of RA were downloaded from the Gene Expression Omnibus (GEO) database. Gene network was constructed, and the genes were classified into different modules using WGCNA. HUB genes in modules related to RA clinical symptoms were analyzed by gene ontology. Subsequently, different data sets of GEO were used to verify the expression profile and diagnostic capacity of the HUB gene [receiver operating characteristic curve (ROC)]. In addition, the expression of HUB gene in RA was verified by real time polymerase chain reaction (RT-PCR) and Western blot, and the relationship between key genes and disease activity score 28 joints (DAS28) was analyzed. Paired-sample t-test and Pearson's correlation analysis was used for statistical analysis. Results:A total of 5 413 differentially expressed genes were filtered. Weighted gene coexpression network was constructed and genes were classified into 23 modules. Among them, the black module is closely related to the clinical symptoms of RA, which contained 346 genes. Enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) signal pathway analysis showed that it was to be enriched in the positive regulation of interleukin 6, interleukin 1 beta secretion, osteoclast differentiation, NOD-like receptor signaling pathway, T helper cell 17 (Th17) cell differentiation and many other pathways closely related to RA. Motile sperm domain-containing protein 2 (MOSPD2) was significantly correlated with clinical symptoms. It was highly expressed in blood monocytes and bone marrow monocytes ( t=2.238, P=0.032; t=3.153, P=0.006), and positively correlated with blood expression in RA joint synovial fluid ( r=0.683, P=0.03). ROC curve analysis determined that MOSPD2 could distinguish RA from the control group (the area under the curve was 0.855 and 0.726) respectively. RT-PCR and Western blotting results showed that MOSPD2 was up-regulated in RA patients ( t=-3.96, P=0.02). MOSPD2 expression levels in blood were positively correlated with DAS28 in RA patients ( r=0.884 6, P=0.046 2). Conclusion:MOSDP2 is closely related to the clinical symptoms of RA patients, and may be one of the targets for the diagnosis and treatment of RA.

Objective To investigate the relationship between the carotid calcification and the prognosis in patients with ischemic stroke. Methods A total of 522 patients with non-cardiac ischemic stroke registered in the Nanjing Stroke Registry Program (NSRP )from December 2009 to October 2012 were enrolled. All patients underwent head and neck CT angiography (CTA). The original data of CT scan were transmitted into the Siemens workstation. Calcium score measurement was performed using the same reconstruction conditions and Agatston calcium score to measure calcification score. The patients were divided into no (0),mild (0

Objective There is little research on the relationship of gene polymorphism of CYP2C19 and clopidogrel response after percutaneous transluminal angioplasty and stenting ( PTAS) in patients with ischemic cerebrovascular disease.The study aimed to investigate the relationship between gene polymorphism and high on-treatment platelet reactivity ( HTPR ) after PTAS and 6 months of regular dual antiplatelet administration in patients. Methods A total of 145 Chinese patients treated with PTAS in our de-partment from January 2011 to March 2014 were enrolled in this study.According to the gene sequencing, patients were divided into wild-type group(CYP2C19*1/*1,69 cases) and mutation group(heterozygous mutation CYP2C19*1/*2 and homozygous mutation CYP2C19*2/*2, 76 cases).Patients received a 100mg/d aspirin and 75mg/d clopidogrel maintenance dose (MD) as dual anti-platelet therapy after PTAS.The clopidogrel inhibition effect was measured by thrombelastography ( TEG) system 6 months after PTAS. Routine cerebral artery digital subtraction angiography was applied to evaluate whether there was restenosis in stent and logistic regres-sion analysis was used to analyze the influential factors of HTPR after PTAS and clopidogrel adminstration. Results After 6 months'regular administration of clopidogrel after PTAS, the platelet adenosine diphosphate ( ADP ) receptor inhibition rates in wild-type group, heterozygous mutation and homozygous mutation group were respectively (58.43 ±21.98)%, (47.80 ±22.93)%, (37.53 ± 21.84)%.The platelet ADP receptor inhibition rate was significantly decreased compared with wild-type group(P=0.001).Carriers of at least one CYP2C19 loss-of-function ( LOF) allele had a higher frequency of clopidogrel HTPR (35.5% vs17.4 % for patients with and without LOF alleles, respectively;P=0.014) .Using multivariate logistic regression analysis, the carriage of CYP2C19 LOF alleles was an independent predictor of the post-procedure HTPR (OR=2.356, 95% CI:1.053-5.272, P=0.037).The rate of ISR was remarkably higher in patients with at least one CYP2C19*2 alleles compared with wild-type patients(11.8%vs 1.4%, P=0.019) . Conclusion In patients with ischemic cerebrovascular disease, the CYP2C19 LOF allele had significant impact on post-procedure clopidogrel HTPR and the prognosis of ISR after PTAS.