Objective To investigate the effects of emodin on the migration and invasion ability and expressions of E-cadherin and Slug of human hepatoma cell lines HepG2; To discuss the possible mechanisms of anti-hepatocellular carcinoma.Methods HepG2 cells were cultured in vitro. The experimental group was treated with emodin at concentrations of 10μmol/L and 20μmol/L as. The negative control group was treated with the same volume of RPMI-1640 medium, while the positive control group was treated with 10μmol/L floxuridine. The cell matrix adhesion assay, wound healing and transwell chamber in vitro invasion assay were used to observe the effects of emodin on HepG2 cell adhesion rate and migration and invasion ability. Western blot analysis was used to observe the changes of expressions of E-cadherin and Slug.Results Compared with the negative control group, emodin inhibited significantly HepG2 cell adhesion rate and migration and invasion ability were in a dose-dependent manner (P<0.05,P<0.01); Western blot analysis showed that the protein expression of E-cadherin increased significantly, and the level of Slug decreased significantly in a dose-dependent manner (P<0.05,P<0.01).Conclusion Emodin can significantly inhibit migration and invasion of HepG2 cells, which mechanism may up-regulate expressions of E-cadherin and down-regulate Slug.

Objective To observe the expression of S100β protein in the traumatic brain injury and investigate its relation to the severity of the TBI patients.Methods To collect 30 volunteer controls,30 patients with traumatic brain injury and 30 patients with trauma expect traumatic brain injury.according Glasgow Coma Scale(GCS),TBI patients were divided into tow groups,the minor group is GCS≥8,the severegroup is GCS<8.ELISA method was used for observing the expression of S100β protein in serum from the controls and patients.Results Within 6 hours after TBI,the concentration of S100β protein increased higher in patients of TBI than the others(P<0.05).The concentration of S100β protein increased higher in the severe group(GCS<8)than the minor group(P<0.05).The higher level of seium S100β protein,the more severe of TBI patients,the higher level of serum S100β protein.Conclusion The serums S100β protein can be a special index for the early diagnosis of TBI,the higher level of it,The more severe of patients.

Aim To evaluate the effect and the mechanism of valdecoxib on Lewis cancer. Methods Western blot was used to detect the expression of VEGF, MMP-2, Bax, Bcl-2 and Caspase-3 in tumor tissue. Flow cytometry was used to observe the effect of valdecoxib on proliferation, apoptosis and the cell cycle distribution. MTT assay was used to observe the lymphocyte transformation rate. Results ① Valdecoxib inhibited the growth of the tumor and increased the survival rate. ② 10~40 mg?kg -1?d -1 Valdecoxib increased the apoptosis rate from 19.1% in control group to 23.1%~29.1%, and did not affect the distribution of cell cycle. ③ The expression level of Bcl-2 was decreased and expression levels of Bax, COX-2, MMP-2, Caspase-3 and VEGF were not affected. ④ Valdecoxib did not affect the weight, the lymphocyte transformation rate, spleen index, and thyrus index of the mice. Conclusion The inhibitory effect of valdecoxib on Lewis tumor is associated with apoptosis.

Objective To evaluate the inhibitory effect of valdecoxib on the growth of the cancer cell lines and involvement of COX-2 in this inhibition. Methods Western blotting and immunocytochemistry were used to detect the expression of COX-2. MTT assay was used to determine inhibitory effect of the drugs on the cell growth. The content of PGE_ 2 in cell medium was determined with PGE_ 2 ELISA kit. Results ①Clone 26 cells expressed high levels of COX-2, whereas BGC-823,HGC-27 and SK-OV-3 cell had no COX-2 expression. ②Valdecoxib inhibited the growth of BGC-823, HGC-27, SK-OV-3 and clone 26 cells, with a IC_ 50 of 110.7, 99.2, 113.3, 117.6 ?mol/L, respectively. ③The inhibitory effect of these drugs on BGC-823 and clone 26 cell was in the descending order of valdecoxib, SC-560 and indomethacin. ④PGE_ 2 did not antagonize the effect of valdecoxib, SC-560 and indomethacin on BGC-823 and clone 26 cells. ⑤The inhibitory effect of valdecoxib and indomethacin on the growth of clone 26 cells was not compatible with that on PGE_ 2 . Conclusion The inhibitory effect of valdecoxib on cell growth is not related to its effect on COX-2.

Objective To study the expression of p27kip1 and Ki67 proteins in various types of gastric polyp and the relationship be-tween gastric polyp and gastric carcinoma (CA:). Methods The expression of p27kip1 and Ki67 proteins were detected in normal gastric mu-cosa, hyperplasie polyp, adenomatons polyp, and carcinoma by S-P immunohistochemical staining. Results In the normal gastric mucosa,hyperplasie polyp, adenomatous polyp and gastric carcinoma, the expression rate of the p27kip1 protein was 90. 00% ,70. 00% ,65.00% and 30. 43%, respectively. The positive rate of p27kip1 protein in gastric cancer was significantly lower than that in other tissues. The rate of Ki67 protein expression in those tissues was 10. 00%, 15.00%, 45.00%, and 73.33%, respectively. The positive rate of Ki67 protein in gastric cancer was significantly higher than that in other tissues. And the positive rate of Ki67 in adenomatous polyp was significantly higher than that in the normal gnstric mucosa and hyperplasic polyp. There were significant differences among them(P <0.05). There was no correlation between the expression of p27kip1 and Ki67 proteins in gastric polyp(rs=-0. 093, P >0. 05). Conclusion Detection of p27kip1 and Ki67 proteins in gastric polyp was very helpful in early diagnosis and prognosis of GC.

Objective To study the methylation status of secreted frizzled-related protein (SFRP) 1 and SFRP2 genes in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) and the relationship between the methylation status of the two genes and the development of HCC.Methods Using methylation-specific polymerase chain reaction (MSP) to detect methylation status of SFRP1 and SFRP2 genes of 45 specimens of HCC tissue and adjacent non-tumorous liver tissue from HCC patients during operations,and 6 normal liver tissues from patients with cholecystolithiasis or hepatic hemangiomas. The data were analyzed by chi-square test and Fisher exact test. Results SFRP1 gene methylation was detected in 28 HCC tissues and 16 adjacent non-tumorous liver tissues,accounted for 62.2% and 35.6%,respectively;and SFRP2 gene methylation was detected in 23 HCC tissues and 13 adjacent non-tumorous liver tissues,accounted for 51.1% and 28.9%,respectively;while no methylation was detected in 6 samples of normal liver tissues. There was no significant difference between the methylation of SFRP1 and SFRP2 genes in HCC tissues and gender,age,HBV serum markers,types of adjacent non-tumorous liver tissues,metastasis and pathological stage (P＞0.05).The abnormal methylation status between SFRP1 and SFRP2 genes was linear correlated in HCC tissues (r=0.381,P=0.01).Conclusion Hypermethylation of SFRP1 and SFRP2 genes frequently occurs in HBV-related HCC,which may be an important molecular biomarker for prediction of hepatocarcinogenesis in the future.

Objective To detect the integration of hepatitis B virus (HBV) DNA in HBVrelated human hepatocellular carcinomas (HCC). Methods Extracted DNA from the liver tissue samples and amplified by nested polymerase chain reaction (PCR) with specially designed U-base primers. According to the known genes and human Alu repeat sequences (Alu repeat) , primers were designed respectively. Integrated clones combined target HBV DNA and the adjacent cell gene sequences were established by PCR and products were sequenced by biotechnology companies.Accurate locations of HBV genes integrated in the human genomes were analyzed by national center for biotechnology information (NCBI) basic local alignment search tool (BLAST) and Map Viewer search. Results In 24 HBsAg positive HCC samples, 15 cases showed the integrations of HBV fragment. And the other 8 samples didn't show any evidence of integration. Among 14 samples with integration, forward insertions of HBV DNA into the host chromosomal DNA were found in 10 samples and reverse insertions were found in 8 samples while both forward and reverse insertions were found in 5 samples. Analysis from viral-cellular junctions suggested that the integrations were all happened with truncated viral DNA and could be in any locus of X gene. Conclusion HBV DNA integration is not distributed evenly throughout the host genome.

OBJECTIVE:To screen antioxidant active components of Schisandra chinensis. METHODS:The orthogonal test was adopted to optimize extraction technology using DPPH free radical scavenging activity(IC50)as index and ethanol volume frac-tion,material-liquid ratio and extraction time as factors,and the verification test were made. The fractions(SC-0,SC-10,SC-30, SC-50,SC-70,SC-95) were made by extracting and purifying S. chinensis with macroporous resin with water and 10%,30%, 50%,70%and 95%ethanol. With IC50 and total antioxidant capacity(determined by ABTS method)as indexes(vitamin C as pos-itive control),the antioxidant active components of S. chinensis were optimized. The contents of 5 kinds of lignan in different posi-tions of S. chinensis were determined by HPLC. RESULTS:The optimal extraction condition of S. chinensis was as follows as 60% ethanol,material-liquid ratio of 1∶14,extracting for 2.0 h. The average IC50 of DPPH free radical scavenging activity was 23.81 mg/ml(RSD=0.52%,n=3)in verification test. SC-0 did not have antioxidant abilities. DPPH free radical scavenging activi-ty of those components (ie. the IC50 value from low to high) were in the following order of positive control>SC-50>SC-30>SC-95>SC-70>SC-10;total antioxidant ability of them were in the following order of SC-50>positive control>SC-30>SC-70>SC-95>SC-10;the contents of 5 types of lignan in different components were in the following order of SC-70>SC-50>SC-95>SC-30. CONCLUSIONS:The antioxidant active component of S. chinensis is 50%ethanol eluate.

BACKGROUND:The homing ability of mesenchymal stem cells is closely associated with the effects of celltransplantation. Clarifying the mechanism of chemotaxis and migration wil contribute to enhance the clinical application of mesenchymal stem cells. OBJECTIVE:To investigate the effect of Cdc42 in the homing of human umbilical cord mesenchymal stem cells. METHODS:First, mesenchymal stem cells were isolated from human umbilical cord, and co-cultured with tumor necrosis factorα, interleukin-1β, and transforming growth factorβ. Western blot assay was used to test the level of Cdc42. Besides, Cdc42 siRNA was synthesized by chemical method to transfect the cells, and cellmigration and adhesion were measured by Transwel and Matrigel separately. Meanwhile, the activity of signal molecule, extracellular regulated protein kinase 1/2, was evaluated by western blot. RESULTS AND CONCLUSION:The results indicated that the inflammation factors induced the highly expression of Cdc42 in human umbilical cord mesenchymal stem cells, almost double level to controls. siRNA notably inhibited the migration and adhesion of human umbilical cord mesenchymal stem cells through Cdc42 down-regulation, and the extracellular regulated protein kinase 1/2 and phosphorylation form were also decreased simultaneously. In a word, we speculate Cdc42 plays a role in the chemotaxis of human umbilical cord mesenchymal stem cells in vitro.

This study was aimed to explore the antitumor effect of volatile oil of X ihuang pill and its immune mecha-nism in order to screen the antitumor active site of Xihuang pill. Among 70 female Wistar rats, 10 rats were random-ly selected as the blank control group; and the other 60 rats was used in the establishment of walker 256 breast can-cer cell tumor-bearing rat models. The model rats were randomly divided into the negative control group (model group), high-dose volatile oil group, middle-dose volatile oil group, low-dose volatile oil group, high-dose Xihuang pill group, and lentinan group (positive control group), with 10 rats in each group. The intragastric administration was given twice a day for 14 days. Blood was taken from the abdominal aorta. Tumor tissues was removed and weighed to calculate the tumor inhibitory rate. ELISA method was used to detect the level of IL-2, IL-6, IL-10, IFN-γ and TGF-β in peripheral blood. The flow cytometry was used to detect the content of CD3+ T cell, CD4+ T cell, CD8+ T cell, and B7-1 cell (CD80). The results showed that the tumor inhibitory rates of volatile oil of high-dose group and middle-dose group were 28.4% and 24.1%, respectively. Compared with the model group, the average level of IL-2 and IFN-γ of volatile oil of high-dose group and middle-dose group and CD3+ T cell, CD8+ T cell, B7-1 cell con-tent were obviously increased (P< 0.05). It was concluded that volatile oil of Xihuang pill had certain antitumor ef-fect, which was one of the antitumor active sites of Xihuang pill. The volatile oil of Xihuang pill upregulates the lev-els of IL-2 and IFN-γ, as well as the contents of CD3+ T cell, CD8+ T cell, B7-1 cell in order to increase the im-mune clearance function of tumor-bearing rats.