Abstract:Objective:To investigate the mechanism that small GTPase Arl6 regulates sonic hedgehog(Shh) signal transduction. Methods:The mRNA expression level of Gli1 and Ptch1,two Shh pathway target genes,were detected in Arl6 knockdown stable cell lines. The localization of Arl6 in the primary cilia was observed under confocal microscope. Ciliogenesis of Arl6 knockdown cells was quantified by counting the percentage of ciliated cells. Results:Knockdown of Arl6 inhibited full activation of the Shh pathway. After treatment with high-concentration Shh medium or Smo agonist Purm,Gli1 mRNA expression was significantly lower in the shArl6 group than in the control group (P < 0.01),and Ptch1 mRNA expression was also decreased (P < 0.05). Arl6 was located in the basal body of the primary cilia, and its absence resulted in defective ciliogenesis. Meanwhile,Shh signaling stimulated the expression of Arl6 (P < 0.05). Conclusion:Knockdown of small GTPase Arl6 inhibits full activation of Shh signaling,which is related to defective ciliogenesis.

Abstract:Objective:To observe the anti-cancer activity of McAb NJ001 against lung adenocarcinoma cells,and explore its molecular mechanism. Methods:We selected SPC-A1 cell lines as research object,used flow cytometry to detect cell apoptosis,and observed morphological changes of SPC-A1 cells by phase contrast microscope. We detected the expressions of apoptosis-related proteins such as caspase-8,caspase-9,caspase-3,PARP,Bax,and Bcl-2 by Western blotting assays. Results:McAb NJ001 induced apoptosis in SPC-A1 cells in a dose-dependent manner. After treatment with NJ001,SPC-A1 cells showed shrinkage,and became round or even fell off,which were typical morphological changes of apoptosis. Results:Western blots showed that after treatment with McAb NJ001,PAPR were cleaved so that caspase-3 expression was decreased,caspase-8 and caspase-9 were activated and their protein expression levels were increased. The expression of pro-apoptotic protein Bax increased while the expression of anti-apoptotic protein Bcl-2 decreased. Conclusion:McAb NJ001 has strong anti-cancer activity due to its apoptosis inducing effect. In addition,caspase activation and up-regulation of Bax/Bcl-2 were involved in NJ001-induced apoptosis.

Abstract:Objective:To identify the novel function of hepatocyte growth factor (HGF) in liver carcinoma cell HCCC-9810 proliferation and the underlying molecular mechanism. Methods:The proliferation of HCCC-9810 treated with HGF was detected by MTT assay and EdU assay. The expression of Akt in HCCC-9810 cells was inhibited by siRNA. Western blot was used to detect the expression of cyclin D1 and the phosphorylation level of Akt. Results:Both 50 ng/ml and 100 ng/ml HGF significantly inhibited the proliferation of HCCC-9810. Western blotting assay showed that HGF downregulated the expression of cyclin D1 and the phosphorylation level of Akt in HCCC-9810 cells. Conclusion:These results suggest that HGF inhibited the proliferation of HCCC-9810 by suppressing the expression of cyclin D1 and the phosphorylation level of Akt. SiRNA interfered the expression of Akt,and blocked HGF-induced downregulation of cyclin D1 expression and inhibition of cell proliferation.

Abstract:Objective:To produce a monoclonal antibody(mAb) against latent membrane protein 2A(LMP2A) with hybridoma technology and characterize its immune specificity. Methods:The spleen cells from BALB/c mice immunized with epitope fusion protein LMP2A were fused with SP2/0 cells to produce a monoclonal antibody against LMP2A. We further determined immuno-specificity of the purified antibody by various immunological assays such as ELISA,Western blotting assays,immunofluorescence,immunohistochemisty and FACS. Results:We successfully produced a hybridoma cell line,which continuously and stably secreted anti-LMP2A mAb. Moreover,this mAb can effectively recognize the transmembrane glycoprotein LMP2A over-expressed in nasopharyngeal carcinoma cells and tissues. Conclusion:We successfully obtained mouse anti-LMP2A mAb that can specifically recognize the transmembrane glycoprotein LMP2A in nasopharyngeal carcinoma cells. It can be potentially applied to early stage clinical diagnosis and some tumor targeted therapy.

Abstract:Objective:To construct and indentify glucocorticoid receptor β (GRβ) shRNA stably transfected human glioma cell lines and investigate the effect of GRβ on glioma cell cycle and proliferation. Methods:The specific siRNA was designed according to human GRβ sequence and transfected into human glioma cells by LipofectamineTM 2000 reagent. Knock down of GRβ by siRNA was measured by RT-PCR and Western blotting assays. The in vitro synthesized siRNA was inserted into shRNA sequence and directionally cloned into eukaryotic expression vector pGPH1/GFP/Neo. After transfection,the stable G0306 cell lines were selected by G418. Western blotting assays,MTS and flow cytometry were applied to detect the effect of GRβ interruption on glioma cell proliferation and cycle,respectively. Results:GRβ siRNA effectively inhibited the expression of GRβ. The inserted sequence of shRNA cloned into eukaryotic expression vector pGPH1/GFP/Neo was confirmed by DNA sequencing. The stable GRβ shRNA cell lines showed growth inhibition in G1 period,and the cell proliferation ability was significantly decreased. Conclusion:Stable GRβ knockdown glioma cell line (G0306) was constructed successfully and GRβ affected glioma cell proliferation,which laid a foundation for further study of the role and mechanism of GRβ in the central nervous system tumors.

Abstract:Objective:The present study was designed to investigate the protective effect of astragaloside IV on mitochondrial injury of aortic endothelial cells from renovascular hypertensive rats. Methods:Two kidney one clip (2K1C) technique was performed to prepare renovascular hypertensive rats. Two weeks after operation,the rats were divided into two groups,the operation group and the astragaloside Ⅳ treatment group[5.0 mg/(kg-d)]. The sham operated rats were used as normal control. After treatment for two weeks,the mitochondrial ultrastructure of thoracic aortic endothelial cells from the rats was observed. The expression of Mn-superoxide dismutase (SOD2) in thoracic aortic endothelium was observed by immunohistochemical staining. Results:Two weeks after operation,the rats showed significant increase of systolic blood pressure compared to that of control rats. The thoracic aortic endothelial cell mitochondria of hypertensive rats showed fracture and disappearance of cristae,swelling and vacuolation as well as decreased mitochondrial matrix electron density. These changes of mitochondrial were ameliorated significantly by astragaloside Ⅳ. In addition,astragaloside Ⅳ augmented the lower expression level of SOD2 in the aortic endothelium from the hypertensive rats compared with that from the normal controls. Conclusion:Astragaloside Ⅳ reverses mitochondrial injury of aortic endothelial cells from renovascular hypertensive rats,which may be linked to its ability to increase SOD2 expression in the aortic endothelium of the rats.

Abstract:Objective:To investigate the protective effects of fasudil,a Rho kinase inhibitor,on the inflammation of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice secondary to sepsis and its probably mechanism in mice. Methods:C57BL/6 mice were randomly assigned to the control group,the LPS group,the LPS+fasudil (10 mg/kg) group and the LPS+dexamethasone (Dex,5 mg/kg) group. Mortality rates of different timing of each group was assessed after the establishment of the model. Mice were sacrificed at 6 h after LPS injection. Serum,bronchoalveolar lavage fluid (BALF) and lung tissues were collected. ELISA was performed to analyze the level of TNF-α,IL-1β and IL-10 in the serum,analyze the cell count and the protein content in BALF,stain with hematoxylin and eosin;test the wet-to-dry (W/D) weight;measure the content of myeloperoxidase (MPO) content in lung tissue by ELISA. Results:Fasudil significantly improved the mortality rates and prolonged the survival time of mice,relieved inflammatory injury and edema of lung tissue,and decreased MPO content. Moreover,fasudil down regulated the expression of TNF-α and IL-1β and up regulated IL-10 in serum. Conclusion:Fasudil effectively relieved LPS-induced ALI,which may be relevant to controlling the inflammatory response in the lungs.

Abstract:Objective:To investigate the effects of high mobility group box 1(HMGB1) antagonist BoxA on seizure onset and hippocampus injury in a rat epilepsy model. Methods:Male SD rats were divided into the sham operation group,the epilepsy model group,the low,middle and high doses of BoxA pretreated epilepsy groups. The rat epilepsy model was made by micro-injection of kainic acid into the hippocampus. Various doses of BoxA were administered to the epilepsy model by intracerebroventricular injection prior to kainic acid injection. The sham operation group were injected with the same volume of normal saline into the hippocampus and lateral ventricle,respectively. We observed the rats’ behavior during seizure and recorded seizure onset time(SOT) to stage Ⅲ as an indicator to assess epileptic susceptibility. The brain pathological change was observed by HE staining. The injury degree of the hippocampus was estimated by NeuN staining as an indicator to assess the damage of brain tissue. The expression of phosphorylated nuclear factor-κB-p65(p-NFκB-p65) was also detected by immunohistochemical staining. Results:SOT in the low,middle and high doses of BoxA pretreated epilepsy group all significantly prolonged compared to the epilepsy group (P < 0.05). Compared to the epilepsy group,hippocampus injury and neuron loss were obviously reduced in the BoxA pretreated epilepsy group (P < 0.05). Although pretreatment with high dose BoxA also prolonged SOT and decreased the intensity of brain injury compared to the middle and low dose BoxA,there were no statistically significant differences. The expressions of p-NFκB-p65 in the middle and high dose BoxA groups were reduced remarkably compared with the EP group (P < 0.05 ),and there was no statistical difference between the EP group and the low dose BoxA group. Conclusion:Administration of HMGB1 antagonist BoxA to epileptic rats could reduce seizure susceptibility and attenuate the injury of brain by inhibit the activation of NFκB,which showed brain protective effect in epilepsy rats.

Abstract:Objective:To investigate the expression of amyloid precursor protein (APP),microtubule-associated protein (Tau),and phosphorylated Tau protein (p-Tau) of offspring mouse hippocampus after perinatal exposure of F0 mice to bisphenol A (BPA) on different stages,and identify critical windows for fetal development stage susceptibility to BPA exposure in male and female mice. Methods:Theprimary cultured hippocampus neurons of C57BL/6J fetal mice receiued with BPA;F0 mice received perinatal subcutaneous injection of BPA to establish the mouse BPA exposure model. Four groups were classified by different exposure stages,including the control,the fetus period (P6-PND0),the lactating period (PND0-PND21) and the combined exposure (P6-PND21) groups. The expressions of APP,p-Tau and Tau in offspring mouse(8-month-old) hippocampus were detected by Western blotting assays. Results:The expressions of APP,p-Tau and Tau protein of hippocampal neurons were increased in a dose dependent manner after incubation with varied concentrations of BPA. BPA obviously increased APP,p-Tau and Tau expressions in the PND0-PND21 and the P6-PND21 group of female mice;for the male mice,the three proteins＇ up-regulation was observed only in the P6-PND21 group. Conclusion:Continuous perinatal exposure to BPA could contribute to the up-regulation of APP,p-Tau and Tau,which might subsequently result in neuronal injury. Moreover,female mice are more susceptible to BPA exposure than male mice.

Abstract:Objective:To investigate the effect and possible mechanism of mammalian target of rapamycin (mTOR) protein and its related pathway on islet β cell proliferation in rats with chronic intermittent hypoxia (CIH). Methods:Twenty-four adult male Sprague-Dawley (SD) rats were randomly divided into two groups (the CIH group and the control group). The CIH group was fed in low oxygen cabins (60 s/N2 and 60 s/air alternately,oxygen concentrations between 5% and 21%,between 8:30~16:30 each day) for 35 days. We detected blood glucose and serum insulin levels by ELISA and calculated the insulin resistance index (HOMA-IR) and β cell insulin secretory function (HOMA-β) of the two groups. Glucose tolerance was evaluated by oral glucose tolerance test. The proliferation of pancreatic β cells was tested by immunofluorescence assay and mTOR and related pathway protein expressions were tested by Western blotting assays. Results:The fasting blood glucose,serum insulin levels and insulin resistance index increased (P < 0.01),while glucose tolerance and pancreatic β-cell function decreased (P < 0.05),in comparison with the control group. The number of ki67 positive β cells increased about five-fold while the mTOR related pathway protein expression of pancreatic tissues was activated. Conclusion:The rats in CIH condition showed insulin resistance,glucose tolerance impairment and pancreatic β-cell dysfunction;meanwhile,the activated mTOR pathway may play an important role in pancreatic β-cell proliferation.

Abstract:Objective:To determine the effects of bone marrow mesenchymal stem cells on mice with endotoxemia. Methods:Mice were divided into the following groups:the control group,the endotoxemia group(the LPS group),the mesenchymal stem cell treatment group (the LPS+MSCs group) and the mesenchymal stem cell group (the MSCs group). The cardiac function of mice was respectively observed at 24 hours and day 7 after LPS administration. ELISA was performed to detect the level of cytokines in the serum,and histological examinations were performed to detect morphological changes of the cardiac muscle,liver,lung and kidney. Results:Compared to the control group,serum IL-1-茁 and TNF-α were increased in the mice of the LPS group,and MSCs treatment decreased serum IL-1-茁 and TNF-α obviously. The cardiac function of the LPS group mice deteriorated,which was rescued by MSCs treatment. Apoptosis of cardiomyocytes and hepatocytes in the mice of the LPS group was increased. Edema was observed in the pulmonary interstitium and alveolus,and ameliorated by MSCs treatment. Conclusion:MSCs inhibit the inflammatory reaction of mice with endotoxemia,improves the cardiac function and reduces the damage of the heart,liver and lung.

Abstract:Objective:To investigate the characteristics of LDH5 expression in tongue squamous cell carcinoma (TSCC) and its relationship with clinicopathological parameters and survival. Methods:The LDH5 protein expression in 63 cases of TSCC and 16 cases of normal tongue mucosa were examined by immunohistochemistry. Based on these patients’ information and their follow-up data,the correlation of LDH5 expression with patients’ clinicopathological characteristics and overall survival were further evaluated by SPSS17.0 software using Fisher exact test and Kaplan-Meier method,respectively. Results:The LDH5 expression in TSCC (33/63,52.38%) was much higher than that in normal tongue (P < 0.001). LDH5 over-expression was found to be associated with tumor size,cervical lymph node metastasis and clinical stages (P < 0.05),but not with gender,age,local invasion and pathological grades(P > 0.05). The overall survival rate in patients with high LDH5 expression was significantly lower than that in patients with low expression as estimated by Kaplan-Meier and Log-rank test(P = 0.006). Conclusion:LDH5 is aberrantly over-expressed in a significant fraction of TSCC patients. Its upregulation is associated with multiple clinicopathological features and patients’ prognosis.

Abstract:Yin Yang -1(YY1) is a member of the zinc finger factor GLI-Krüppel family. Objective:To investigate the gene expression and the function of YY1 in normal esophageal tissues and esophageal squamous carcinoma. Methods:The protein expression of YY1 in 88 esophageal squamous cell carcinomas,39 tumor adjacent tissues and 15 normal esophageal specimens was determined by immunohistochemistry (IHC). Proliferation and growth changes of esophageal squamous carcinoma cells were measured by MTT and colony formation. YY1 and p21 protein expressions were determined by Western blotting assays. TE-1 cell invasion and migration were determined by cell scratch experiment and Transwell invasive assays. Results:Compared to normal esophageal and tumor adjacent tissues,the expression of YY1 was significantly higher in esophageal squamous cell carcinomas. Furthermore,TE-1 cells overexpressing of YY1 significantly suppressed cell growth and downregulation of YY1 promoted cell growth. Meanwhile,a change of YY1 target gene p21 (p21WAF1/Cip1) was found. TE-1 cells overexpressing YY1 promoted cell invasion and migration. Conclusion:The expression of YY1 was significantly increased and regulated the growth,invasion and migration ability of esophageal squamous cells. Therefore,YY1 is expected to be a molecular marker in the diagnosis and development of esophageal squamous cell carcinoma,and also a potential target for gene therapy of esophagus cancer.

Abstract:Objective:To detect the expression of miR-30c/snail and the role of miR-30c in hepatocelluar carcinoma (HCC) invasion. Methods:mRNA expression of miR-30c/snail in HCC samples and cell lines were determined by real-time PCR,while protein expression of snail in HCC tissues and cells were detected by immunohistochemistry and Western blotting assays,respectively. Moreover,invasion of HCC cells was analyzed after transfection with miR-30c mimic or inhibitor. Results:We found that the miR-30c levels were obviously decreased in HCC with vascular metastasis compared with non-metastatic tissues,and the same results were found in the highly metastatic cell line MHCC-97H. Transfection with miR-30c mimic decreased snail expression and reduced the invasion of MHCC-97H cells;however,transfection with miR-30c inhibitor increased the protein level of snail and promoted the invasion of HepG2 cells. Conclusion:Upregulation of miR-30c expression attenuates the invasion of HCC by directly inhibiting snail. A new therapeutic strategy based on miR-30c may prove to be beneficial for HCC patients with invasion or metastasis.

Abstract:Objective:To observe the expression of interleukin 22(IL-22), interleukin 22 receptor 1(IL-22R1) and signaling and transcriptional activation factor 3(STAT3) in the different stages of malignant transformation of gastric cancer, and to illustrate the effect of IL-22 in the process. Methods:The tissues from patients with chronic gastritis (15 cases),gastric precancerous lesions(15 cases) and gastric carcinoma (15 cases) were collected and examined for IL-22,IL-22R1 and p-STAT3 expression by immunohistochemistry and real-time PCR. Results:Immunohistochemical analysis showed that IL-22 was mainly located in the cytoplasm of gastric epithelial cells and infiltrated immune cells. IL-22R1 was mainly expressed in the cytoplasm of gastric epithelial cells, while the location of p-STAT3 was mainly in the nucleus of gastric epithelial cells. The expression of IL-22, IL-22R1 and p-STAT3 in precancerous lesions and gastric carcinoma were significantly more than that in chronic gastritis. At the same time, IL-22,IL-22R1 and p-STAT3 were significantly increased in gastric carcinoma compared with precancerous lesions(P < 0.001). The expression of IL-22 and IL-22R1 mRNA were higher in precancerous lesions and gastric carcinoma than in chronic gastritis, and were higher in gastric carcinoma than in precancerous lesions(P < 0.05). Conclusion:In the cells of precancerous lesions and gastric carcinoma,IL-22 can be secreted by infiltrated immune cells and can also be self-secreted by atypical glandular epithelium.

Abstract:Objective:To investigate the significance and different expressions of microRNA-210 in astrocytic tumors and oligodendroglial tumors. Methods:Tissue samples were divided into three groups including the normal brain tissue group,the oligodendroglial tumor group and the astrocytic tumor group. The astrocytic tumors were divided into four groups including group A[pilocytic astrocytoma (WHO grade I)],B[diffuse astrocytomas (WHO grade II)],C[anaplastic astrocytomas(WHO grade III)]and D[glioblastoma (GBM,WHO grade IV)]. The expression of miR-210 was examined by real-time quantitative RT-PCR. The correlation between the expression of miR-210 and the astrocytic tumors was analyzed by Spearman correlation test. Results:The expression level order of miR-210 was group D> group C> group B and group A. There was statistically significant difference between groups except group A and B (P < 0.001). The grade of astrocytic tumor and the expression of miR-210 had positive correlation. However,the expression level of miR-210 in anaplastic astrocytomas was lower than in oligodendroglial tumor,and was low in oligodendroglial tumor (P < 0.005). Conclusion:Since expression of miR-210 showed significant differences in different origins and different grades,it can be used as an auxiliary identification method for the histopathologic examination of glioma from different origins,or as a biomarker for the observation of the progression of astrocytic tumor as well.

Abstract:Objective:To investigate the independent and combined effects of smoking status,overall obesity and abdominal obesity on risk of hypertension. Methods:Participants were recruited under the framework of the community diagnosis investigation from Zhonglou district of Changzhou in 2012. In this investigation,a total of 7 437 subjects aged above 18 years were included in this study. Logistic regression model was performed to analyze WC,BMI and smoking as well as the relationship between their interaction and hypertension. We tested an interaction on an additive scale by using an Excel spreadsheet set up by Andersson to calculate relative excess risk due to interaction(RERI),attributable proportion due to interaction(AP),the synergy index(SI),and then estimate the confidence interval(CI). Results:After adjustment for sex,age,alcohol consumption,family history of high blood pressure,occupation,diet and etc,hypertension OR (95%CI) was 2.75 (2.46~3.12) in overweight or obese subjects,and 2.41 (2.13~2.71) in abdominal obese subjects,compared with normal subjects. OR(95%CI) was 1.37 (1.16~1.63) for smokers compared to never smoking subjects. Compared to never smoking and non-abdominal obese subjects,OR (95%CI) was 3.66 (3.07~4.36) in abdominal obese smokers,2.01 (1.78~2.03) in never smoking and abdominal obese subjects,and 1.47 (1.26~1.71) in non-abdominal obese smokers. RERI was 1.18(0.57~1.79),AP was 0.322 and SI was 1.80(1.35~2.38). Conclusion:Smokers have a significantly higher hypertension risk than never smoking subjects. Moreover,this study further demonstrates an additive interaction of smoking and abdominal obesity on hypertension risk.

Abstract:Objective:To determine miR-126 serum levels in children with congenital heart disease(CHD). Methods:A total of 54 children with CHD who received treatment in Department of Cardiology from January 2013 to June 2013 were enrolled. There were 40 children with left-right shunt acyanotic CHD(the ACHD group),including 15 children with heart failure(ACHDⅠ)and 25 children without heart failure(ACHDⅡ). There were 14 children with right-left shunt cyanotic CHD(the CCHD group)and 20 healthy children(the control group). The serum miR-126 expression level was measured to analyze the correlation between the serum miR-126 of neonates with CHD and heart failure. Results:Compared with normal controls,the expression levels of miR-126 in the ACHD group and the CCHD group were significantly down-regulated(P < 0.001). The expression levels of miR-126 in the CCHD group were significantly lower than those in the ACHD group(P < 0.05 and P < 0.001,respectively). MiR-126 expression levels were lower in the ACHDⅠ group those in the ACHDⅡ group(P < 0.001). Conclusion:The expression levels of serum miR-126 in children with CHD were significantly decreased and correlated with the severity and degree of impaired cardiac function,indicating that miR-126 may be involved in the pathological process of CHD,and it may serve as an independent evaluation indicator for the diagnosis of CHD.

Abstract:Objective:To discuss the influence of pregnancy complications on late onset fetal growth restriction (FGR). Methods:A total of 130 patients with FGR and 130 randomly selected normal pregnant women were collected from Nanjing Maternity and Child Health Care Hospital during 2013. The incidences of hypertensive disorder complicating pregnancy,anemia,gestational thyroid hypofunction,gestational diabetes mellitus (GDM) and intrahepatic cholestasis of pregnancy (ICP) in the two groups were compared. Meanwhile,folates of red blood cell in the two groups were also determined for comparative analysis. According to the different symptomatic treatment and nutrition status in FGR fetuses,130 cases of FGR were divided into two subgroups,48 cases in the no-intervention group and 82 cases in the intervention group. The rate reaching the standard of neonatal body weight,premature birth,fetal distress,and neonatal asphyxia,neonatal pneumonia between the two subgroups was compared. Results:Compared with the control group,the occurrence rates of hypertensive disorder complicating pregnancy,anemia and gestational thyroid hypofunction were significantly higher in the FGR group (P < 0.05). However,the difference in the occurrence rates of GDM and ICP between the two groups was not significant (P > 0.05). The level of red blood cell folate in the FGR group was significantly lower than that in the control group (P < 0.05). The rate reaching the standard of normal neonatal body weight in the intervention group was significantly increased compared to that in the none-intervention group (P < 0.05). The incidences of premature birth,fetal distress,neonatal asphyxia and neonatal pneumonia in the intervention group were also lower than those in the none-intervention group (P < 0.05). Conclusion:Hypertensive disorder complicating pregnancy is the most important factor causing fetal growth restriction. Meanwhile, anemia, lack of folic acid and gestational thyroid hypofunction and some other factors also directly affect the growth and development of the fetus. Interventional treatment can significantly improve pregnancy outcome according to etiology.

Abstract:Objective:To explore the incidence of cervical Modic changes and related clinical factors of cervical spondylitic myelopathy(CSM) of patients requiring surgical intervention by MRI image. Methods:A total of 133 patients with CSM were collected from our hospital between May,2009 and March,2010. Flexion extension radiographs and preoperative MRI were collected. We evaluated cervical Modic changes and measured space available for the cord(SAC). Then,we measured total angle of motion(TAM) and segmental angle of motion(SAM) on cervical flexion-extension radiographs. Rank sum test was performed to analyze the clinical relevant factors. Results:Modic change incidence was 51.1% from patients with cervical myelopathy requiring surgical intervention. In patients who were over 60-year-old and with increased extension activity and more severe spinal stenosis, the incidence of Modic change was higher. The length of medical history and sex were not correlated to Modic change. Conclusion:Severe spinal stenosis and the increment of SAM on partial cervical flexion-extension radiograph due to aging and protrusion of intervertebral disc were intensively correlated with cervical Modic changes.

Abstract:Objective:To evaluate the difference and correlation of activated clotting time(ACT) measured between Celite i-STAT- Ⅱ and clinical classical measurement method-Medtronic ACT Ⅱ. Methods:Sixty patients underwent elective cardiac surgery. We compared ACT values measured by Medtronic ACT-Ⅱ and Celite i-STAT- Ⅱ. Blood samples were obtained before establishment of extracorporeal circulation,after heparinization and protamine neutralization,respectively,and then measured by the two methods. Linear regression and Pearson correlation analysis were performed to assess the consistency of the two measured data. Bland-Altman plot was performed to evaluate bias. Results:Celite i-STAT-ACT value was 85 to 913 seconds and Medtronic ACT-Ⅱ value was 99 to 980 seconds,R2 = 0.96,regression equation:y = 7.86 + 0.86x,Pearson correlation coefficient:r = 0.98(P < 0.001). ACT value ranged from 480 to 999 seconds and had linear correlation,R2 = 0.65,regression equation:y = 108.82 + 0.72x,Pearson correlation coefficient:r = 0.80(P < 0.001). Bland-Altman plot showed that the results of the two methods had good consistency. Conclusion:In ardiac operation under extracorporeal circulation,ACT values measured by Celite i-STAT- ACT and Medtronic ACT-Ⅱ are acceptable clinically.

Abstract:Objective:To compare clinical efficacy and safety between intravenous glucocorticoid (IVGC) pulse therapy and oral route in the management of Graves’ ophthalmopathy. Methods:All randomized controlled trials (RCTs) comparing IVGC with oral route for the management of Graves’ ophthalmopathy were identified by manual and computer retrieval. All related data were extracted. Meta analysis was conducted using the statistical software RevMan 5.2 on the basis of strict quality evaluation with the methods recommended by the Cochrane Collaboration. Results:Fifty-four papers were found and 5 RCTs involving 256 patients (134 patients in the IVGC group and 131 patients in the oral route group) were included. The meta analysis showed that the IVGC group was associated with significant improvement of clinical activity score (CAS) compared with the oral route group (WMD = 0.82,95%CI:0.61~1.03,P < 0.001),proptosis (WMD = 0.62,95%CI:0.11~1.12,P = 0.02),lid width (WMD = 0.72,95%CI:0.14~1.30,P = 0.01),but the reduction of diplopia showed no difference between the two groups (OR = 1.51,95%CI:0.77~2.95,P = 0.23). IVGC had lower incidence of side effects than the oral route (OR=0.16,95%CI:0.08~0.30,P < 0.001). Conclusion:Compared with the oral route,IVGC pulse therapy showed better efficacy and safety in the management of Graves’ ophthalmopathy.

Abstract:Objective:To construct a eukaryotic expression system of human anti-TROP2 antibody IgG by using human anti-TROP2 Fab antibody gene as template,so as to study its inhibition of pancreatic cancer cell proliferation. Methods:The recombinant expression vector of human anti-TROP2 antibody IgG,named as pWS-anti-TROP2,was established by amplifying the heavy chain and light chain genes of human anti-TROP2 antibody IgG,respectively. pWS-anti-TROP2 plasmids were transfected into CHO dhfr- cell lines,and the monoclonal cell strains with high antibody expression were harvested by MTX screening. Then,human anti-TROP2 antibody IgG was obtained using Protein G affinity purification. The identification and immunological characteristics of the human anti-TROP2 antibody IgG were analyzed by several methods,including sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE),Westent blotting assay,enzyme-linked immuno sorbent assay(ELISA),flow cytometry method(FCM) and immuno fluorescence assay. The inhibition function of human anti-TROP2 antibody IgG in proliferation of BxPC3 cells was analyzed by MTT assay. Results:The eukaryotic expression system of human anti-TROP2 antibody IgG was constructed successfully. Purified human anti-TROP2 antibody IgG was obtained. The molecular weight of light and heavy chains of human anti-TROP2 antibody IgG are consistent with expected results,the antibodies could bind to TROP2 protein specifically,and the antibody titer reached 1∶6 400. MTT assay demonstrated that human anti-TROP2 antibody IgG played a significant pole in inhibiting BxPC3 cell proliferation,and the inhibition ratio was gradually increased with prolonged time and increased antibody dose. Conclusion:In the study,the eukaryotic expression system of human anti-TROP2 antibody IgG was established successfully,and it was proved that the antibody could recognize native TROP2 protein on pancreatic cancer cell surface specificially and play an important role in inhibiting the proliferation of pancreatic cancer cells.

Abstract:Objective:To investigate the effects of high fat diet and estrogen on cardiac structure and glucolipid metabolism. Methods:The rats were divided into 3 groups,the high-fat diet group,the ovariectomy and high-fat diet group and the ovariectomy and estrogen injection group. The change of body weight,visceral fat weight,blood glucose and blood insulin levels,serum estradiol and serum lipid levels were measured. Echocardiography was performed to evaluate the related indexes of cardiac structure. The histomorphological changes in hearts were observed under microscope by using hematoxylin-eosin (HE) staining. Results:Estrogen was negatively correlated with the degree of obesity;it can reduce fasting blood glucose and improve insulin resistance;it also decreased serum total cholesterol,protected myocardial cells by mitigating the degeneration and necrosis. Conclusion:Estrogen can effectively improve obesity-induced insulin resistance and abnormal lipid metabolism and protect heart structure.

Abstract:Objective:To determine if the cardiac output (CO) measured by transesophageal echocardiography (TEE) through the left ventricular outflow tract (LVOT) is consistent with that measured by pulmonary artery catheter (PAC). The correlation between left ventricular ejection fraction (LVEF),left ventricular fractional area change (LVFAC) and right ventricular ejection fraction (RVEF) was analyzed. Methods:Twelve patients with ASA Ⅱ~Ⅲ(NYHA Ⅱ or Ⅲ),aged 18~70 years,weighing 46~72 kg and undergoing coronary artery bypass grafting were studied. Anesthesia induction and intraoperative maintenance were performed by intravenous anesthesia. After tracheal intubation,the Swan-Ganz catheter and TEE probe were placed. The data were measured and recorded after tracheal intubation (T0),15 (T1),30 (T2) and 60 (T2) min after termination of cardiopulmonary bypass (CPB) or finished vascular anastomosis in off-pump CABG. Statistical analysis was performed by Bland-Altman plot and Pearson correlation coefficient method. Results:COPAC was (4.82 ± 1.32) L/min and COLVOT was (4.57 ± 1.30)L/min. COLVOT was highly correlated with COPAC (r = 0.655,P < 0.001). The bias between COPAC and COLVOT was 0.28 L/min (95%CI:-0.04~0.60 L/min),and limits of agreement were -1.90~2.46 L/min. LVEF was positively correlated with LVFAC (r = 0.662,P < 0.001),while it was not significantly correlated with RVEF (r = -0.218,P > 0.001). The bias between LVEF and LVFAC was 15.36% (95%CI:12.46%~18.29%),and limits of agreement was 4.68%~35.43%,while that between LVEF and RVEF was 34.40% (95%CI:29.69%~39.10%),and limits of agreement was -2.01%~66.78%. Conclusion:The results showed that CO measured by TEE through the LVOT was significantly correlated but poorly consistent with that measured by Swan-Ganz PAC. Both methods for CO measurement cannot replace each other. Moreover,there was an obviously bias but a good correlation between LVFAC and LVFAC.

Abstract:Objective:To investigate the effect of data perturbation in microarray on selecting differentially expressed genes by false discovery rate (FDR). Methods:A total of 1 991 DNA microarray data of colon cancer were afforded random perturbation of different error limits based on a computer simulation. Every perturbation comprised 1 000 random simulations. The differentially expressed genes were selected from data with and without perturbation,respectively,by adaptive linear step-up (ALSU),a method of FDR. The repetition rates between both results were compared. The effect of each gene sort order was analyzed by data perturbation. Results:The single average and overall average repetition rates of differentially expressed genes both decreased with increasing data perturbation. The more significant differentially expressed the genes,the less they were affected by perturbation. When the error limit was less than or equal to 50%,the overall average repetition rate of differentially expressed genes decreased with increasing data perturbation linearly. For each 1% increase of perturbation error limit,the overall average repetition rate decreased approximately by 1.85%. The higher the perturbation error limit,the greater the fluctuation the gene sort order had. Conclusion:Data perturbation is a reason why differentially expressed genes exhibit low repeatability;the effect of data perturbation on selecting differentially expressed genes can be quantitatively investigated by using computer simulation.