AIM: To assess the effect of raloxifene, a selective estrogen modulator, on the transcriptional activity of estrogen responsive element (ERE) in vascular endothelial cells. METHODS: Vascular endothelial cells were transfected with ERE-TK-Luc reporter plasmid and PRL-SV40 control plasmid via FuGENE 6. The activities of firefly luciferase and renilla luciferase were measured with dual-luciferase reporter assay system. The transcriptional activity in transfected cells treated with raloxifene was compared with that in untreated cells. Furthermore, raloxifene was used to cotreat the cells with estradiol (E_2) and the influence of raloxifene to the effect of E_2 was assessed. RESULTS: Compared to untreated cells, the luciferase activity in cells treated with raloxifene decreased and showed significance at concentration of 10~(-7)mol/L (P

Objective To investigate the protective effect of caffeine citrate on white matter damage in 2-day-old neonatal rats induced by postnatal infection.Methods Forty-eight 2-day-old SD rats were randomly divided into the control group (group A,n =16),lipopolysaccharide(LPS) infection group (group B,n =16),and caffeine citrate intervention group (group C,n =16) according to the random table method.The newborn rats of group B and C were continuously injected LPS 0.6 mg/kg intraperitoneally for 5 days from 2 days old,and the newborn rats of group A were continuously injected by an equal volume of 9 g/L saline intraperitoneally.Group C was continuously injected by caffeine citrate 10 mg/kg intraperitoneally for 7 days from 4 days old;equal volume of 9 g/L saline was injected into group A and B for 7 days continuously.At 12 days old,8 rats of each group were sacrificed randomly to evaluate the expression of myelin basic protein(MBP) subcortical white matter by immunohistochemical method.Both sides of hippocampus of the rest 8 rats of each group were taken out in ice surface rapidly.The left hippocampus was used to detect the expression of MBP and Caspase-3 by Western blot method,and the right hippocampus was used to evaluate the MBP and Caspase-3 protein level by quantitative real-time PCR(qPCR) method.Results The mean integral optical density (IOD) of subcortical MBP positive expression in group A,group B and group C were 132.64 ± 1.94,102.43 ± 2.12,114.25 ± 2.04,and the difference was statistically significant among 3 groups (F =22.912,P ＜ 0.05).The relative expression levels of MBP mRNA of 3 groups in hippocampus were 0.79 ± 0.01,0.39 ± 0.03,0.55 ± 0.02,and the difference was statistically significant among 3 groups (F =18.584,P ＜ 0.05).The relative expression levels of MBP protein of 3 groups in hippocampus were 0.64 ± 0.03,0.31 ± 0.03,0.51 ± 0.05,and the difference was statistically significant among 3 groups (F =25.780,P ＜ 0.05).The relative expression levels of Caspase-3 mRNA of 3 groups in hippocampus were 0.34 ± 0.02,0.74 ± 0.03,0.57 ± 0.04,and the difference was statistically significant among 3 groups (F =6.105,P ＜ 0.05).The relative expression of Caspase-3 protein of 3 groups in hippocampus were 0.11 ± 0.03,0.36 ± 0.02,0.23 ± 0.03,and the difference was statistically significant among 3 groups (F =40.541,P ＜ 0.05).Conclusions Caffeine citrate has showed protective effect on white matter damage in neonatal rats of 2 days old induced by postnatal infection.The mechanism may be related to inhibition of inflammatory response and reducing apoptosis.

Objective To elucidate the roles of high mobility group box-1 protein (HMGB1) and myeloperoxidase (MPO) deficiency in evaluating coronary stenosis and the vulnerability of atherosclerotic plaques in patients with coronary heart disease.Methods Totally 50 patients with acute myocardial infarction (AMI),50 patients with unstable angina pectoris (UAP),50 patients with stable angina pectoris (SAP) and 30 patients without coronary heart disease underwent the study.Coronary arteriography (CAG) and intravascular ultrasound (IVUS) were performed to analyze coronary stenosis and plaque characteristics and then gensini score was calculated.Concentrations of HMGB1,MPO and hypersensitive C reactive protein (hsC-RP) were measured by means of enzymelinked-immonosorbent assay (ELISA).Results Concentrations of HMGB1,MPO and hsC-RP were significantly higher in AMI and UAP patients than in SAP paticnts (all P＜ 0.01).IVUS showed that 51.3 ％ (20/39) AMI patients,46.7％ (43/92) UAP patients had soft lipid-rich plaqucs,while 52.9％(46/87) SAP patients had fibrous plaques,only 17.2％(15/87) had soft plaques.AMI and UAP patients had larger plaque burden and vascular remodeling index than did the SAP patients (both P＜0.01).In AMI group,HMGB1 and MPO levels were correlated well with gensini score and remodeling index measured by IVUS,respectively(r=0.54,0.48,allP＜0.05),while in UAP group,HMGB1 and MPO levels were correlated well with gensini score and plaque burden measured by IVUS,respectively(r=0.43,0.56,all P＜0.05).Conclusions HMGB1 and MPO are positively correlated with coronary stenosis,which can be used to predict the severity of ACS.HMGB1 and MPO are associated closely with plaque vulnerability and rupture.

AIM: To assess the effect of estrogen on the gene expression of caveolin-1 in rat vascular smooth muscle cells (VSMCs). METHODS: Wistar rats were ovariectomized and subjected to subcutaneous implantation of placebo pellets (OVX+V group) or estradiol pellets (OVX+E group). 2 weeks after implantation, the expression of caveolin-1 gene in endothelium-denuded aortic tissue was examined by RT-PCR. Furthermore, Northern blotting was used to analyze the mRNA expression of caveolin-1 in cultured rat VSMCs. RESULTS: RT-PCR showed that expression of caveolin-1 gene was significantly higher in OVX+E group than that in OVX+V group. Northern blot analysis showed that the mRNA expression of caveolin-1 was higher in VSMCs pretreated with 17?-estradiol (17?-E 2) than that in VSMCs without 17?-E 2 pretreatment. CONCLUSION: Estrogen up-regulates the gene expression of caveolin-1 in the vascular wall, partially indicating the cardiovascular effect of estrogen. [

Objective To investigate the effect of raloxifene on the mRNA expression of endothelin-1 in vascular endothelial cells and assess the role of estrogen receptor. Methods Bovine carotid endothelial cells were pretreated with 10nM raloxifene for 24 hours, then total RNA was harvested and Northern blotting was performed to investigate the effect of raloxifene on the mRNA expression of endothelin-1. Furthermore, estrogen receptor inhibitor, 100nM ICI 182 780 was used to pretreat the cells together with raloxifene and the expression of endothelin-1 was observed too. Results The mRNA expression of endothelin-1 in bovine carotid endothelial cells was inhibited significantly by pretreatment with raloxifene and this effect could be blocked by ICI182 780 (0.16?0.05 vs 0.39?0.07, P

From September 2002 to May 2006, five patients ( age range, 24 to 46; mean, 32) with tibial defect underwent transplantation of free latissmus dorsi osteocutaneous flap using bridge-like vascular anastomosis. All the osteocutaneous flaps survived without any serious complications, and tibial defects were improved completely. After 1.5 to 4.5 years' follow-up ( mean, 2.6), no remarkable dysfunction was found at donor sites, and local injury was reduced. This study indicates that transplantation of free latissmus dorsi osteocutaneous flap using bridge-like vascular anastomosis might be useful in leg reconstructive surgery if only one vessel is remained.

Eight newborn screening quality indicators were developed though expert discussing organized by National Center for Clinical Laboratories according to NBS quality management in USA .These indicators can not only be used to evaluate the performance throughout critical aspects of pre -examination, examination and post-examination processes for routine tests , but also be applied to monitor a series of processes from specimen collecting to results interpretation in newborn screening .Meanwhile, NCCL can use these quality indicators to carry out External Quality Assessment programs in order that the total service quality of newborn screening can be improved.

Objective To analyze the status of reference intervals (RIs) of complete blood cell count (CBC) in China and compare them with the published standards.Methods The RIs information of CBC (containing the items of WBC,RBC,Hb,HCT,MCV,MCH,MCHC and PLT) were collected by the External Quality Assessment (EQA)software from laboratories participated in the Second Complete Blood Cell Count EQA Program in 2013,which including the sources of RIs,the lower and upper limits of RIs and the validation information of RIs.The sources and validation information of RIs were analyzed by Microsoft Excel 2007 using all data except those from maternal and child care service center,children's hospital and corporations.The outliers of limits of RIs were excluded according to the International Federation of Clinical Chemistry(IFCC)guidelines,then data were analyzed by SPSS 13.0.The mean of the upper and lower limits of the RIs were compared with the published standards by the t-test.Results Totally 299 (57.72％) of 518 laboratories reported were using RIs according to National Clinical Laboratory Operation Rules,while 112(26.62％)labs were using RIs from instrument/reagent manufacturer instructions,411 (80.00％)labs did not validate the RIs they used,of which 258 (50.00％) did not know how to validate RIs.The deviation of the maximum and the minimum ranged from 16％ to 70％.More than one third laboratories did not separate RIs of RBC,Hb and HCT according to the gender.The upper and lower limits of WBC and MCV of most laboratories were higher,while those of RBC(male/female),PLT and Hb(male) were lower than that from the published standard.The RIs of MCH was narrower than that from the published standard.Meanwhile,HCT,MCHC (male/female) and Hb (female) were near the upper and lower limits of published standards.Conclusions At present,the RIs of CBC used in laboratories is chaotic,which is significantly different from the published standard.On the basis of standardization,the RIs of published standard result from a multi-center study in Chinese people should be preferred.

Objective To study the correlations between genetic polymorphisms of TNF-α as well as IL-6 and susceptibility to colorectal cancer among Chinese Han people in Shandong province.Methods Single nucleotide polymorphisms (SNPs) of TNF-α-238G/A,-308G/A and IL-6-174G/C,-572G/C,-597G/A in 490 patients with colorectal cancer were analyzed by using gene chip.Concentrations of TNF-α and IL-6 in serum samples were measured by ELISA.A case-control study was conducted to analyze the correlations between SNPs of TNF-α-238G/A,-308G/A as well as IL-6-174G/C,-572G/C,-597G/A and susceptibility to colorectal cancer.Chi-square test or t test was used for statistical analysis.Relative risks were estimated based on the values of odds ratio (OR) and 95% confidence interval (95%CI).Results The frequency of TNF-α-308AA in patients with colorectal cancer was significantly higher than that in healthy subjects (x2 =6.15, P<0.05, OR=2.08, 95%CI=1.17-3.71), while the frequency of IL-6-572CC in patients with colorectal cancer was significantly lower than that in healthy subjects (x2 =4.97, P<0.05, OR=0.73, 95%CI=0.55-0.96).The frequency of TNF-α-308AA in patients with colon cancer (OR=2.31, 95%CI=1.17-4.55), tubular adenocarcinoma (OR=2.32, 95%CI=1.28-4.21), high (OR=2.05, 95%CI=1.01-4.15) or moderately differentiated adenocarcinoma (OR=5.88, 95%CI=1.79-19.30) was significantly higher than that in healthy subjects.The levels of serum TNF-α in TNF-α-308AA carriers with colorectal cancer were significantly higher than those in TNF-α-308G carriers with colorectal cancer (t=2.13, P<0.05) as well as those in healthy TNF-α-308AA carriers (t=2.13, P<0.05).The levels of serum IL-6 in colorectal cancer group were significantly higher than those in control group (t=6.74, P<0.001).Conclusion The SNPs of TNF-α-308 and IL-6-572 are associated with the occurrence and development of colorectal cancer in Chinese Han people in Shandong province.

Objective To compare the effect of treating cricopharyngeal achalasia in stroke survivors using transnasal or transoral balloon dilatation.Methods Thirty stroke survivors with cricopharyngeal achalasia were randomly divided into a transnasal and a transoral balloon dilatation group (group N and group O),each of 15.Both groups were given routine swallowing rehabilitation training as well as the transnasal or transoral balloon dilatation.Their heart rate was monitored during the dilatation.Nasal bleeding,mucous membrane swelling and pain were also observed.Their swallowing function was evaluated using the Fujishima Ichiro swallowing efficacy score (FISE) and videofluoroscopy (VFSS) before and after the intervention.Results After the treatment,the average FISE and VFSS scores of both groups had improved significantly comnpared to before the treatment but there were no significant differences between the groups.During the treatment,the average heart rate of group O increased significantly less than that of group N.The treatment acceptance of group O was 98.2％,significantly higher than that of group N (80.1％).One case of mucosal bleeding was observed in group O,and laryngeal edema occurred significantly less often than in group N (9 cases vs.7).The average pain score was also significantly lower in group O.Conclusions Balloon dilatation facilitates swallowing among stroke survivors with cricopharyngeal achalasia.The transoral approach can help to reduce the occurrence of complications such as mucosal bleeding,laryngeal edema and pain,and has better patient acceptance.