Objective To investigate the effect and molecular mechanism of Andrographolide (AD) on matrix metalloproteinase-9 (MMP-9) expression in human colon cancer H 3255 cells. Methods Human lung cancer H3255 cell line weve cultured in vitro, and treated with 1.0, 3.0, 5.0μmol/L AD for 24 h, untreated cells was used as blank control. Cell viability, cell migration and cell invasion were analyzed by MTT assay, scratch healing assay and transwell membrane assay, respectably. Expression of MMP-9 mRNA was analyzed by RT-PCR. Protein expression and phosphorylation of Akt were detected by Western blot. Activity of NF-κB and MMP-9 were analyzed by luciferase reporter assay. Results AD could significantly reduced H 3255 cells invasion and migration without affecting the viability of cells, as demonstrated by scratch healing and transwell membrane assay. Furthermore, Western blot and RT-PCR results showed that AD could markedly inhibited MMP-9 activity and its expression in both protein and mRNA levels. AD could attenuated Akt’s phosphorylation and the activity of NF-κB. Moreover, LY 294002, an inhibitor of PI3 K, could significantly inhibited NF-κB transcriptional activity and MMP-9 expression. In addition, different concentrations of AD could inhibit the promote activity of MMP-9. Conclusion AD was a potential anti-invasive agent by inhibiting MMP-9 involved in PI3 K/NF-κB pathways.

Objective Serum glycated albumin (GA) is associated with the levels of blood glucose and short term glycemic fluctuation. The aim of this study was to investigate the associations between GA and arterial stiffness in a health screening population. Method A total of 11 014 participants (men, 56.2%;mean age, 46.8±7.8 years) who attended physical examination were invited and had completed standardized questionnaires. Serum GA and homocysteine (HCY), brachial-ankle pulse wave velocity (baPWV), and central systolic blood pressure (cSBP) were measured. A series of single and multiple factor analyses and stratified analysis were performed. Result The mean level of serum GA was lower in men (13.69 ± 2.69) than in women (13.82 ± 1.92) (t=2.78, P=0.005). The values of baPWV (men: F=53.9, P for trend<0.001;women:F=37.7, P for trend<0.001) and cSBP (men:F=10.3, P for trend<0.001;women:F=13.5, P for trend<0.001) increased with quartiles of GA levels in both men and women. After adjustment for age, history of hypertension, diabetes, family history of diabetes, smoking status, body mass index, waist circumference, blood pressure, blood lipids, kidney function, fasting glucose, postprandial glucose, and hemoglobin A1c, GA was positively associated with baPWV (βestimate, 0.019;95%CI:0.002-0.036;P=0.032) and cSBP (βestimate, 0.251; 95% CI: 0.107-0.394; P=0.001) in men. Stratified analysis also showed that the positive associations of GA with baPWV and cSBP remained significant in men both with and without diabetes or hypertension (P<0.05). However, the association between GA and HCY was not significant in multifactor analysis and stratified analysis. The above results were similar in women. Conclusion Serum GA levels were positively associated with baPWV and cSBP not only in patients with diabetes or hypertension but also in participants without diabetes or hypertension after adjusting for conventional factors and glucose markers. The findings suggest that GA may help detect the development of arterial stiffness in the health screening population.

Objective To study the mechanism of FTY720 on type Ι diabetic rats. Methods The typeⅠdiabetes rat model was established by feeding male SD rats with high energy urea and injecting into the abdominal cavity with low dose cephalosporins (STZ, 30 mg/kg). In the following, the treated rats were divided into two groups: model group and FTY720 group. Another group of untreated rats was assigned as normal control group. FTY720 group was given 1 mg/kg FTY720, the normal control group and model group was given the equal amount of water. Results The cardiac function of FTY720 group was recovered markeyly. FTY720 activated the expression of vascular endothelial cells S1P1 , diabetes and reduced the expessions of S1P3 and PKCβ Ⅱand it restored the migration ability of diabetes cardiovascular endothelial cell , as well as the abnormally elevated cardiovascular endothelial cells induced by high sugar permeability. Conclusion The S1P1 and S1P3 cut is an important reaction pathway to the complications of diabetes and cardiovascular dysfunction. FTY720 may reduce the damage to core blood vessels caused by diabetes , and pathological angiogenesis which functionally depends on the PKCβⅡ signaling pathways. Therefore, FTY720 may provide a potential therapy for cardiovascular function impaired by diabetes.

BACKGROUND:The levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) are increased during infectious brain edema, and are positively relevant to the degree of brain damage. However, whether TNF-α can enhance blood brain barrier (BBB) permeability remains unclear, especially in vitro.OBJECTIVE: To understand the changes and possible mechanism of the BBB permeability induced by TNF-α in vitro.DESIGN: Randomized controlled cell model study in vitro.SETTING:Department of Pediatrics, Xiangya Hospital, Central South University; Department of Biochemistry, Xiangya Medical College, Central South University.MATERIALS: Twenty 7-day-old healthy Sprague-Dawley rats, of clean grade and either gender, were provided by the Animal Center, Xiangya Hospital, Central South University. TNF-α was purchased from sigma Company; DMEM fluid medium and fetal bovine serum were purchased from Hyclone Company; Y-27632 was purchased from Alexis Company,and rabbit anti-human factor Ⅷ -related antigen was purchased from Zymed Company; Mouse anti-rat glial fibrillary acidic protein (GFAP) was purchased from Neomarkers. Other biochemical reagents were imported (Sigma Company).METHODS: This experiment was carried out in the Xiangya Hospital, Central South University between March 2004 and April 2005. Brain microvascular endothelial cells and astrocytes were co-cultured 10 days to set up rat models of BBB in vitro. Then, the cells were divided into 4 groups: model group(BBB models were prepared), TNF-α group ( BBB model incubated with 0.01 g/L TNF-α for 5 hours), Y-27632 pretreated group ( BBB model incubated with 30 μmol/L Y-27632 for 1 hour before 0.01g/L TNF-α challenge ) and Y-27632 control group (BBB models only incubated with Y-27632 as those in the Y-27632 pretreated group). The effect of TNF-α on BBB permeability was observed by detecting the 125 I -BSA, which passed through the inserts at each time point (30,60,120 and 240 minutes) using .γradioimmunoassay counter.MAIN OUTCOME MEASURES: The 125 I -BSA, which passed through the inserts in rat models of BBB at different time points after intervention.RESULTS: The 125 I -BSA, which passed through the inserts in rat models of BBB, was all significantly higher in the TNF-α group than in the other groups at 30, 60, 120 and 240 minutes after intervention, respectively (P ＜ 0.01), and reached the peak at 240 minutes; The 125 I -BSA, which passed through the inserts, was lower in the Y-27632 pre-treated group than in the TNF-α group at 30 and 60 minutes after intervention (P＜ 0.01). There was also significant difference in 125 I -BSA permeation between Y-27632 pretreated group and Y-27632 control group after 120 minutes (P ＜ 0.05).CONCLUSION: TNF-α can increase BBB permeability, and Y-27632 pretreatment can early reverse the effect of TNF-α on BBB permeability.

Objective To observe the role of heme oxygenase (HO)-1 on the protective effect of C-phycocyanin (CPC) on doxorubicin (DOX)-induced myocardial cells injury by Nvf2/HO-1 pathway. Methods 60 SD rats were randomly divided into control group, DOX group, CPC group and tin protoporphyrin IX (SnPP, an inhibitor of HO-1) group. The control group was injected with normal saline injection,while the DOX group was administrated with doxorubicin by intraperitoneal injection in a cumulative dose of 15 mg/kg for two weeks. For the CPC rats, 20, 40 and 60 mg/kg of CPC was administrated. The level of creatine kinase (CK) and lactate dehydrogenase (LDH) were detected, and the activity of HO-1 and caspase-3 were also examined. Expression of HO-1 and activation of Nrf 2 were detected by Western blot. Results Compared with control group, serum levels of CK, LDH and Caspase-3 activity in DOX group were significantly increased(P<0.05), but HO-1 in cardiac muscle was only increased slightly. upregulation. Treatment with CPC could significantly ameliorated the CK, LDH and Caspase-3 activity, and markedly induce HO-1 expression and its activity. The reduction of CK, LDH and Caspase-3 activity by CPC could be reversed by treatment of the HO-1 inhibitor, SnPP. Furthermore, CPC sould also induce Nrf 2 activation. Conclusion The protective effect of CPC on doxorubicin-induced myocardial cells in jury via Nrf 2 induced HO-1 HO-1 expression.

Objective:Cholangioduodenal reflux is still an unresolved problem in interposed jejunal choledochoduodenostomy (IJC) yet.To settle this problem,we designed IJC with distal suspending (IJCDS).Methods:IJCDS is to insert and suspend the distal end of the 25 cm-ling interposed jejunum to the duodenal cavity,which is about 2.5 cm long.The pressure of the biliary tract and the biochemical composition or the bile were measured during and after the operation and the results were compared.Barium meal and cholangiography through T tube were performed after the operation.Follow-up surveys were also done.Results:The pressure or the biliary tract decreased distinctively after the operation.Barium meals found no cholangioduodenal reflux although the patients had changed many postures.Cholangiography through T tube showed parisaltic or the interposed jejunum and the contrast medium was continuously pushed into the duodenum.Noreflux cholangitis and stone recurrence were found in the follow-up survey in 14 patients.Conclusion:IJCDS has the powerful function on preventing cholangioduodenal reflux.On the other hand,it avods the shortcomings of other reflux-preventing operative pattern.So it had the good prospect to extend its clinical usage.

Objective To assess the value of chest computer tomography (CT) and serum tumor marker in lung cancer screening among health check-up populations.Methods A total of 88 596 adults who had a medical check-up (including chest CT,chest DR,and tumor biomarkers test) in our center were enrolled in this investigation.Clinical diagnosis of lung cancer was confirmed by three professional doctors.Receiver operating characteristic (ROC) curve was used to assess the relationship between serum level of tumor biomarkers and lung cancer.Results The average detection rate of lung cancer was 59.8/100 000 in our center during 2007 and 2011 (men 42.9/100 000,women 92.2/100 000).Rate of positive results of chest CT was 94.3％,which was approximately 2 times of chest DR.Moreover,carcinoembryonic antigen (CEA) testing was effective in early diagnosis of lung cancer with higher sensitivity (73.6％) and specificity (57.1％).Conclusion This study suggests that health checkup centers should pay more attention to lung cancer screening in high risk populations,and new technologies might be used to facilitate the early diagnosis of lung cancer.

Objective To analyze the clinical information of a family with one patient who have systemic lupus erythematosus (SLE) and Fabry disease,as well as the enzymatic activity and gene mutation in these family members.Methods Clinical characteristics were collected from the proband and her family members.Peripheral blood samples from three members of this family were collected and the enzymatic activity was measured by fluorimetrie substrate assay.Genomic DNA was extracted from one male member with significantly decreased enzyme activity,the 7 exons and their flanking introns of GLA gene were amplified by PCR and directly sequenced.Results The enzyme activity of two family members was significantly decreased,the genetic analysis of the male member revealed a missense mutation in exon 2:c.334C＞T (CGC＞TGC)( p.R112C ).Family members except the proband had no definite evidence to support the presence of SLE.Conclusion The coexistence of SLE and Fabry disease is extremely rare.Immunological test,enzymatic activity and gene mutation analysis seem to be helpful for the differential diagnosis.

Objective To assess the effect of lifestyle intervention on risk factors of chronic disease.Methods A total of 400 adults with high-risk of chronic disease received 3 months' lifestyle intervention.The effect of lifestyle intervention was then estimated.Results A total of 387 adults took physical examination.After lifestyle intervention,body mass index,waist circumference,blood pressure,glucose,glyceride,total cholesterol,and high density liopportein were significant decreased (t values were 27.50,19.01,7.46,6.56,5.29,7.74,7.27 and-7.64,respectively; all P＜0.05); however,low density liopportein showed no significant difference (t=0.73,P=0.469).A total of 373 adults had lifestyle and cardiovascular risk estimated.The scores of lifestyle and physical exercise showed statistically significant difference before and after lifestyle intervention (x2 values were 48.405 and 50.778,respectively; both P＜0.05).However,diet,alcohol drinking and cigarette smoking showed no significant difference (x2 values were 0.087,0.112 and 1.410,respectively; all P＞0.05).Moreover,the estimated cardiovascular risk showed significant difference (x2=10.284,P＜0.05).Conclusion Lifestyle intervention could be an effective tool for those with higher risk of chronic disease.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.