Two-dimensional electrophoresis performed on a microfluidic chip could reduce the consumption of sample and reagents, realizing high-throughput analysis in a very short time. During the isoelectric focusing ( IEF) separation, the buffer for the two different separation methods should be kept from being mixed. After the IEF, the protein was transferred from the IEF channel into the second dimension channels. Based on the pseudo valve structure, this paper reported a method to deposit a titanium dioxide membrane at the interface of two dimensional channels, which enhanced the performance of the pseudo valve. The IEF electrophoresis and SDS-polyacrylamide gel electrophoresis ( SDS-PAGE ) procedures were characterized respectively. The total separation took 10-15 min, and the protein's migration rate was linear with the migration time, and inversely proportional to the logarithm of protein molecular weight. Based on the above results, the differential two-dimensional electrophoresis was performed to test the deviation between different measurements.

Objective Using a short-term in vivo laboratory model of murine liver cancer induced with diethylnitrosamine (DEN),we inspected the impact of 5% C.Chrysantha leaves and 5% concentrations on induction of liver cancer with DEN.Methods Laboratory Winstar rats,all being male and SPF grade,were administered with the liver cancer initiator DEN(200?g/kg,ip,once only),then with murine food containing 0.015% 2-AAF (2-Acetaminoflueren) for 2 weeks,following an excision of the augmented parts as a selective promoting program that speeded up the proliferation of precancerous foci from the initiated hepatocytes.On the beginning of this experiment,5% C.Chrysantha leaves and 5% concentration-contained food were given to the trail rats; in contrast,normal food to the control.Rats were killed 3 days after feeding of 2-AAF-contained food.Targeting part of liver were collected for ?-glutamyltranspeptidase (?-GT) staining,and numbers and areas of?-GT positive foci per cm2 of the liver area were calculated using a software designed by us.Results Numbers and areas of?-GT positive foci of the trail are significantly less than those of the control.Conclusion 5% C.Chrysantha leaves and 5% concentrations inhibit the DEN induction to liver cancer.

Objective To investigate the relationships between the severity of non-alcoholic fatty liver disease (NAFLD) and metabolic syndrome (MS).Methods One hundred and twenty-seven cases of NAFLD patients were selected from March 2011 to August 2012 in the First Hospital Affiliated to Fujian Medical University,of them,61 patients with mild NAFLD,45 patients with moderate and 21 patients with severe.And 21 cases without NAFLD were selected as control group during the same hospitalized period.All objects received the measures of height,body weight,waist circumference (WC),blood pressure; Liver ultrasonic examination,the examination of fasting plasma glucose,blood fat and hepatic function detections were also handed by special people.Results The proportion of overweight in the control group and the three NAFLD subgroups were 57.1％ (12/21),88.5％ (54/61),95.6％ (43/45) and 100％ (21/21) respectively (x2 =18.376,P ＜0.001) ;The proportion of the obesity in control group and the three NAFLD subgroups were 19.0％ (4/21),44.3％ (27/61),64.4％ (29/45) and 71.4％ (15/21) respectively(x2 =16.440,P =0.001).The proportion of the metabolic syndrome of the control group and the three NAFLD subgroups were 14.3％ (3/21),45.9％(28/61),71.1％ (32/45) and 71.4％ (15/21) respectively (x2 =22.637,P ＜ 0.05).All three subgroups of NAFLD were higher than the control group (x2 =6.641,P ＜ 0.05 ; x2 =18.562,P ＜ 0.05 ; x2 =14.000,P ＜0.05,respectively).The severity of NAFLD was positively correlated with BMI,WC,TG,FBG,SBP,and DBP (r =0.467,0.503,0.386,0.369,0.279,0.295,P ＜ 0.01),and negatively correlated with HDL-C (r =-0.209,P ＜0.05).Conclusion The severity of NAFLD had significant correlations with metabolic syndrome's components.

Objective:To study the effect of Ginkgo biloba extract (EGb761) on metabolism of aflatoxin B_1(AFB_1) in Wistar rats. Methods:Seventy one Wistar rats were divided into three groups at random: group A (AFB_1 group), group B (AFB_1+EGb761 group), and group C (control group). The rats in groups A and B were given AFB_1(intraperitoneal injection, 100-200 μg/ kg body weight, 1-3 times/week). The rats in group B were fed the food containing EGb761 while the rats in groups A and C were given normal food. Blood samples were collected and liver biopsy was performed on the 14th, 28th and 42nd week. All the rats were sacrificed at the 64th week. The incidence of hepatoma was observed. The hepatic phase Ⅰ drug-metabolizing enzyme CYP450 and phase Ⅱ enzyme GST were detected by spectrometry. The serum AFB_1-lysine adduct was determined by high performance liquid chromatography (HPLC). The expression of 8-hydroxydeoxyguanosine(8-OHdG) was measured by immunohistochemistry. Results:The incidence of hepatocellular carcinoma (HCC) in group B was significantly lower than that in group A (26.92% vs 76.00%,P<0.001). No hepatocellular carcinoma developed in group C. EGb761 had no effects on the activities of CYP450 and GST in rat liver tissues. The level of AFB_1-lysine adduct reached the peak (4 356.01 pg/mg albumin) at the 14th week in group A. EGb761 significantly inhibited the formation of AFB_1-lysine adducts in serum by 13.07% at the 14th week (P＝0.033), and 73.63% at the 42nd week (P＝0.002). The expression of 8-OHdG protein in rat liver tissues in group B was significantly lower than that in group A at the 28th, 42nd, and 64th week (P<0.05). Conclusion:The main mechanism underlying the effect of EGb761 in blocking hepatogenesis induced by AFB_1 may not be fully related with its influence on the activity of liver phase Ⅰ and phase Ⅱ metabolizing enzymes. EGb761 inhibites the production of AFB_1-lysine addcuts, decreases the expression of 8-OHdG protein, and finally alleviates the DNA oxidative injury, which may be one of the mechanisms for the effects of EGb761 in inhibiting or delaying hepatogenesis induced by AFB_1.

Risk factors of hyperamylasemia in 152 diabetic ketoacidosis(DKA)patients who had no acute pancreatitis were examined.Serum levels of hemodiastase,natrium,kalium,chlorine,calcium,phosphonium,creatinine,carbon dioxide combining power,osmotic pressure,blood glucose,blood lipids and uric acid were measured.With hemodiastase and other 14 factors as independent variables,the multiple linear regression analysis was performed and the partial regression coefficient was estimated.Blood glucose,serum natrium,osmotic pressure and triglyceride entered the regression equation,and their partial coefficients were 10.26,10.35,2.21 and 8.00,respectively.The constant quantitv was -2162.06.Compared with amylase-normal group,hyperamylasemia group showed statistically significant difference in blood glucose,serum natrium,osmotic pressure and triglyceride levels.Higher blood glucose and triglyceride might be the primary causes of hyperamylasemia.

[ ABSTRACT] AIM:To investigate the effect of silencing cell division cycle 25a ( CDC25a) gene on the prolifera-tion of human hepatoma HepG2 cells.METHODS:CDC25a gene in human hepatoma HepG2 cells was silenced by RNA interference.Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells.Western blotting was applied to detect the expression of CDC25a at protein level.In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells.RESULTS:The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P<0.05).The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P<0.05).The cell proliferation in silence group was lower than that in negative control group and normal control group ( P<0.05) .The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase.CONCLUSION:Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effec-tively inhibits the CDC25a gene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25a gene may be a key target for the treatment of liver cancer.

Objective To explore the basic ingredients of the tree shrew’ s( Tupaia belangeri) milk and compare with the dairy ingredients of other milks.Methods We select ten seed tree shrews after delivery ( 1 ~21 ) d with lactation mother tree shrews, and use artificial passive breastfeeding method let the young tree shrews suck breast milk,we took the milk from the young tree shrews in the stomach, directly using aseptic operation with a syringe immediately, once every two days, for consecutive three to five times, and a total of 18 mL milk was taken from each seed tree shrew.Then the milk was detected according to the national standard method for component testing.Results The total solid content of the tree shrew’ s milk was 43.63%, including 26.01%of fat, 10.41%of protein, 0.45% of lactose and 0.99%of ash content.Compared with cow's milk, the tree shrew’ s milk contained 3.36 times of total solid contents, 1.24 times of ash, 2.74 times of protein, 6.67 times of fat, and 0.09 times of lactose.Compare with baby formula milk, the tree shrew’ s milk contained 1.44 times of total solid contents, 0.20 times of ash, 0.58 times of protein, 1.53 times of fat, and 0.06 times of lactose.The trace mineral composition of the tree shrew’ s milk showed that the calcium, phosphorus, potassium, sodium, magnesium, and iron contents were 1.83 times, 2.73 times, 1.25 times, 1.93 times, 1.28 times, and 1.48 times higher than those in the cow's milk, and were 0.66 times, 0.85 times, 0.34 times, 0.26 times, 0.85 times, 0.24 times lower than those in baby formula milk.Conclusions The main nutrients of tree shrew’ s milk is of high fat, high protein and low sugar, and it can provide a basis for tree shrews artificial brood and breeding work.

Objective To investigate the relationship between waist-to-height ratio and metabolic syndrome,in order to identify the optimal cut-off point of waist-to-height ratio for predicting metabolic syndrome.Methods In this cross-sectional study,we recruited 343 people who received physical examination in First Hospital of Quanzhou between January 2012 and June 2014,and collected the information of their waist circumference,height,weight,blood pressure,laboratory test results (including fasting blood glucose,2-hour glucose after oral glucose tolerance test,triglyceride,high-density lipoprotein cholesterol) and visceral fat area assessed by computed tomography.Then a receiver operating characteristic (ROC) curve analysis was used to estimate the optimal cut-off points of waist-to-height ratio for the prediction of metabolic syndrome.Results Among the 343 people,there were 195 metabolic syndrome patients,the prevalence rate was 56.8％,which was 70.2％ in men (127/181) and 42.0％ in women (68/162).In ROC curve analysis,the area under the curve of waist-to-height ratio for the prediction of metabolic syndrome was 0.664 for men,and 0.673 for women.The optimal cut-off point of waist-to-height ratio for predicting metabolic syndrome was 0.543 0 (sensitivity 88.2％,specificity 44.4％) for men,and 0.568 3 (sensitivity 86.8％,specificity 46.8％).Conclusion The optimal cut-off point of waist-to-height ratio for predicting metabolic syndrome in Quanzhou population is 0.543 0 for men and 0.568 3 for women.

Objective:To test the expression of Minichromosome maintenance complex component 7(MCM7) protein in hepato-cellular carcinoma(HCC) of different species including human, rat and tree shrew (tupaia) by cross-species oncogenomics approach, and to investigate the relationship between the expression of MCM7 and the development of hepatocellular carcinoma and its clinical significance. Methods:Western blot and Immunohistochemistry were applied to detect the expression levels of MCM7 protein in HCC tissues,corresponding HCC-adjacent liver tissues and normal liver tissues collected from different species including human, rat and tree shrew, respectively. The clinicopathologic factors were also analyzed with the results of Immunohistochemistry. Results:Western blot analysis showed that the expression of MCM7 protein in HCC tissues of human and rat were higher than that in corresponding HCC-ad-jacent liver tissues and normal liver tissues, respectively and significantly (P<0.05). However, the expression of MCM7 protein in HCC tissues of tree shrew were also higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, but no significant difference was found among three types of tissues (P>0.05).There was also no significant difference between HCC-adjacent liver tis-sues and normal liver tissues in three species (P>0.05). Immunohistochemical analysis showed that MCM7 protein was mainly ex-pressed in nucleus of HCC cells, and the positive rate of MCM7 protein in HCC tissues of human, rat and tree shrew were significantly higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, respectively (P<0.05). However, no significant difference was found between HCC-adjacent liver tissues and normal liver tissues (P>0.05). Moreover, the protein level of MCM7 was intimately related to patient's HCC stage, extrahepatic metastases and postoperative recurrence (P<0.05). Conclusion:MCM7 protein might play a pivotal role in hepatocarcinogenesis. In addition, it was probably related to patient's HCC stage, extrahepatic metastases and postoperative recurrence. It seems very likely that MCM7 may be applied as a new molecular target in HCC prevention and treat-ment.

Objective:To search the marker of micrometastatic hepatocellular carcinoma (HCC).Methods:Peripheral blood samples were obtained from 65 patients with hepatocellular carcinoma,21 non HCC malignant tumors,22 chronic hepatitis B or cirrhosis,and 21 normal healthy volunteers.To identify hepatocellular carcinoma cells in peripheral blood, liver specific alpha fetoprotein(AFP)mRNA was amplified from total RNA extracted from whole blood by nested reverse transcription polymerase chain reaction (Nested RT PCR).Results:AFP mRNA was not detected in the normal healthy volunteers and patients with non HCC malignant tumors.The presence of AFP mRNA in patients with HCC(44/65,67.7%)was higher than those with chronic hepatitis B or cirrhosis(2/22,9.1%, P