Objective To investigate the changes of CA125 level with intra-abdominal tumor(epithelial ovarian cancer ,fallopian tube cancer and peritoneal cancer ) before and after chemotherapy serum and their effect on the prognosis of patients and their the clinical predictive value .Methods 174 cases(107 cases of epithelial ovarian cancer ,35 cases of fallopian tube cancer ,and 32 cases of peritoneal cancer) with peritoneal cancer patients were selected from January 2007 to January 2008 .The percentage decrease of ser-um CA125 were calculated after 3 courses of chemotherapy and CA125 decreased level of sensitivity with chemotherapy were ana-lyzed .The different levels of CA125 of median survival time and median survival time were compared .Results After chemothera-py ,CA125 decreased ≥ 75% in 42 cases ,decreased from 51% to 75% in 62 cases ,decreased from 25% to 50% in 32 cases ,and de-creased less than 25% in 38 cases .In CA125 decreased ≥ 75% group ,the chemotherapy effects was better than the other groups (P<0 .05) .Univariate analysis showed that the decrease proportion of CA 125 was positively correlated with chemotherapy effect (r=0 .396 ,P=0 .000) .Using the Kaplan-Meier method ,the patient's survival rate and median survival time were caculated and it showed that the 5 years survival and median survival time were significantly better in group CA 125 decline ≥ 75% than the other groups(P<0 .05) .Univariate analysis shows that the CA125 decline proportion of patients and long term efficacy was positively correlated(r=0 .412 ,P=0 .000) .The COX risk model analysis showed that FIGO stage ,CA125 level and the effect of chemothera-py were independent prognostic risk factors .Conclusion The level of CA125 is the independent risk factor of epithelial ovarian cancer ,fallopian tube cancer and peritoneal cancer prognosis .CA125 decline proportion could be used to know the effect of chemo-therapy and long term treatment ,and be the prognosis indicators for patients with intra abdominal tumor .

To screen genes in endothelial cells resulted from responding to lipopolysaccharide (LPS), mRNA was extracted from both untreated human umbilical endothelial cells (HUVEC) following and HUVEC which were treated with LPS for 6 hours. cDNAs of both populations were synthesized to generate cDNA libraries by suppression subtractive hybridization (SSH). The libraries were then screened with colony dot blots. Positive clones were sequenced and BLAST analysed. The results showed differential genes included 3 novel genes and 22 known genes. The 3 novel genes were confirmed by Northern blotting analysis. These 22 known genes were involved in the regulation of proinflammatory response, cell apoptosis, cytoskeleton, signal transduction and energy metabolism. These results suggest that SSH is an effective technique to detect differential gene expression in HUVEC, which may be helpful to evaluate molecular mechanisms of endothelial injury induced by LPS and provide potential therapeutic targets for LPS related disturbances.

To clone the full length cDNA sequence of a novel expression sequence tag ST55 (GenBank Accession No. BI121646) from human umbilical vein endothelial cells stimulated by lipopolysaccharide, rapid amplification of cDNA ends (RACE) was used to extend the 3′and 5′ends of ST55 to obtain the full length cDNA sequence according to the known ST55 sequence. The cDNA was identified by Northern blot and analyzed with bioinformatics. A novel human full length cDNA was cloned and named endothelial overexpressed lipopolysaccharide associated factor 1( EOLA1 ) (GenBank Accession No. AY074889). This gene was located at chromosomal Xq27.3 and encodes a protein EOLA1 composed of 158 amino acids. Our data show that EOLA1 is a novel human gene associated with activated endothelial cells, and may play a role in intercellular signal transduction.

Objective To study the subcellular localization and the tissue expression of EOLA1(endothelial-overexpressed lipopolysaccharide-associated factor 1). Methods The fusion protein EOLA1-EGFP expressed vector was constructed and transfected into endothelial cells. After 24 hours posttransfection, the subcellular localization of EOLA1 was detected by laser-scanning microscopy. The tissue-specific distribution of EOLA1 was assessed with Multiple Tissue Northern Blots. Results The expression of EOLA1 was tissue-specific in various human tissues. With human multiple tissue Northern blot analysis, it was shown that EOLA1 could express in the heart, skeletal muscle, kidney, liver, placenta, colon, spleen, small intestine, but did not in the brain, lung, thymus, and peripheral blood leukocyte. EOLA1 was mainly localized in cytoplasm and could move into the nucleus. Conclusion EOLA1 is one of intercellular proteins and may play a role in intercellular signal transduction.

Objective To construct small interfering RNA(siRNA) expression vectors targeting EOLA1 (Endothelial-overexpressed lipopolysaccharide-associated factor 1) gene, and to assay their effects on the expression of EOLA1. Methods GFP-EOLA1 fusion protein expressive vector pEGFP-N2/EOLA1 was constructed and transfected into ECV304 cells. The transfected cells were cultured in M199 containing G418 (400?g/ml) for 5 weeks to screen the cell line stably expressing GFP-EOLA1 fusion protein. For interfering EOLA1 target gene, complementary oligonucleotides were desiqned and synthesized at different sites. The oligonucleotides were inserted into pSinencer3.1/H1 plasmid. Then, the vector was transfected into the ECV304 cells stably expressing GFP-EOLA1 fusion protein and the inhibiting affection to target gene EOLA1 was identitied by the observation of the green fluorescence in transfected cells with inverted fluorescent microscope and Western immunoblot assay. Results The ECV304 cell line stably expressing GFP-EOLA1 fusion protein was constructed, and the siRNA vector of EOLA1 can knockdown EOLA1 gene expression specifically. Conclusion The siRNA vectors specifically interfering EOLA1 expression were constructed successfully, which would be useful in the study of function of EOLA1 gene.

Objective To screen and analyze genes up regulated in human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide (LPS). Methods Suppression subtractive hybridization (SSH) was performed between the unstimulated HUVEC(driver) and HUVEC stimulated with LPS(tester) to generate subtractive cDNA library. The library was screened with colony dot hybridization to further verify the differentially expressed cDNA clones. Positive clones were sequenced and BLAST analyzed. The 3 novel cDNA sequences were verified by RT PCR. Results Twenty five up regulated genes related to inflammation, cellular cytoskeletal rearrangement, cellular proliferation and apoptosis, intercellular message transduction, and 3 new expression sequence tags (EST) were acquired. RT PCR indicated the expression of the new ESTs only in HUVEC stimulated by LPS. Conclusion SSH is a powerful technique of high sensitivity for the detection and clone of up regulated gene expressed in HUVEC stimulated by LPS, which may be helpful to clarify the mechanism of endothelial cells activation stimulated by LPS.

Objective To identify the promoter sequence of endothelial-overexpressed lipopolysaccharideassociated factor 1 ( EOLA1) gene and to elucidate the molecular mechanisms controlling EOLA1 expression. Methods A DNA fragment containing 1 723 bp 5' upstream of the EOLA1 gene and the transcription start site was generated by polymerase chain reaction and then cloned into a luciferase reporter gene vector,pGL3-basic. The relative luciferase activities driven by this 5'-upstream fragment and a series of deletion mutants were measured in transiently transfected human ECV304 cells,respectively. At last,the 1 723 bp upstream of the EOLA1 gene was analyzed online with Cluster Buster. Results A fragment 785 bp upstream of the EOLA1 coding region was sufficient to promote transcription. Further deletion analysis of the 785 bp fragment indicated that a 68 bp element from-738 to -676 was important for EOLA1 transcription in ECV304 cells. The 1 723 bp sequence contains binding sites for Sp1 and Myf. Conclusion We map the EOLA1 promoter by deletion analysis and reveal that the proximal region ( -738 to -676 bp) ,which contains binding sites for Sp1 and Myf,is essential for human EOLA1 promoter activity in ECV304 cells.

Objective To construct the ?-gal reporter genes containing the 5′-end flanking of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) gene in different sequence lengths and identify the sequence, which regulates the gene expression of EOLA1 by the ?-gal analysis system. Methods The target sequences were amplified by the method of genome walker, and were inserted into the upstream of ?-gal gene located in the ?-gal enhancer vector by the directional clone technique respectively; the regulative sequence was identified by analyzing the ?-gal activities of reconstructed plasmid in ECV304 cells. Results The regions, containing 2 659 bp and 1 951 bp upstreaming from exon 1, significantly stimulated the reporter gene activity as compared with that of the ?-gal control vector in transfected cells. But the region, containing 361 bp upstreaming from exon 1, did not stimulate the reporter gene activity. Conclusion There is an up-regulative element of gene transcription in the region of -361 to -1 951 bp in EOLA1 gene upstream.

Objective:To investigate the correlation of the relative cerebral blood flow (rCBF) of three dimensional arterial spin labeling (3D ASL) with vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in glioma. Methods:Fifty-three glio-ma patients confirmed by pathology were subjected to conventional, enhanced MR and 3D ASL imaging before operation to deter-mine VEGF expression and MVD levels in each patient. The correlations of rCBF with VEGF expression and MVD in glioma were evaluat-ed, respectively. Results:rCBF was noted to be positively correlated to VEGF expression and MVD in glioma. The rs were 0.728 (VEGF) and 0.620 (MVD), respectively (P<0.05). Conclusion:The positive correlation of rCBF with VEGF expression and MVD in glioma implied that 3D ASL is beneficial for evaluating microvessel angiogenesis in glioma prior to surgery. This finding is significant for developing clin-ical treatment plans and for assessing patient prognoses.

Objective To evaluate the role of mTOR in spinal cord in the development of diabetic neuropathic pain in rats.Methods Sixty adult male Sprague-Dawley rats,aged 2 months,weighing 180-220 g,were used in the study.Forty-five rats among them were chosen randomly and diabetes mellitus was induced by intraperitoneal streptozotocin (STZ) 60 mg/kg and confirmed by blood glucose ＞ 16.7 mmol/L on day 3 after STZ injection.The left 15 rats received intraperitoneal injection of the equal volume of citric acid-sodium citrate buffer and served as normal control group (group C).Paw withdrawal threshold to von Frey filament stimulation was measured in the right hind paw before STZ injection and on 3,6,9,12,15,18,and 21 days after STZ injection.The diabetic rats with mechanical pain threshold decreasing by more than 50％ of the baseline were allocated to diabetic neuropathic pain group (group DP),and by less than 25 ％ of the baseline were allocated to diabetic non-neuropathic pain group (group NP).The rats were sacrificed at 21 days after STZ injection,and their lumbar enlargements of the spinal cord were removed for determination of the expression of mTOR and phosphorylated mTOR (p-mTOR) by Western blot.Results The expression of mTOR was significantly up-regulated in DP and NP groups when compared with group C (P ＜ 0.05),the expression of p-mTOR was up-regulated in DP group,and no significant change was found in the expression of p-mTOR in group NP (P ＞ 0.05).Compared with group NP,the expression of p-mTOR was significantly up-regulated (P ＜ 0.05),and no significant change was found in the expression of mTOR in group DP (P ＞ 0.05).Conclusion Activation of mTOR in the spinal cord is involved in the development of diabetic neuropathic pain in rats.