Objective To investigate whether SUMO-1 enhances the apoptosis induced by wild-type p53 plasmid transfection in HepG2 cells. Methods The HepG2 cells were transfected respectively or simultaneouly with the following expressional plasmids as pcDNA3-wtp53(pwtp53,including human wild-type p53 gene),pCMV-HDM1B(pMDM2,including HDM2 gene, homologous gene as murine double minute gene 2),pcDNA3-His6-SUMO-1(pSUMO-1 ,including small ubiquitin-like modifier-1 gene)and plasmid pcDNA3.The proteins expressed in cells were detected by means of Western blotting and the apoptosis rates of cells were measured by flow cytometry. Results The protein bands of p53 and MDM2 could be seen in cells transfected with pwtp53 and pMDM2. Meanwhile,the relative larger molecular weight bands were also seen in cells transfected with pSUMO-1 which represented the p53 and MDM2 protein modification by SUMO-1. Merely the trace of p53 protein was detected in cells not transfected with any plasmid or only transfected with empty plasmid and pSUMO-1. In cells transfected with pwtp53 and pwtp53+pSUMO-1,the apoptosis rates were (16.79?1.62)% and (18.15?1.36)%. When transfected with pwtp53+pMDM2,the rate decreased to (5.17?1.23)%. The apoptosis rate would come up again to (14.06?1.84)% after transfected with pwtp53+pMDM2+pSUMO-1 and the difference of rates were significant compared to the cells transfected with pwtp53+pMDM2 (P

To clone the murine alpha-fetoprotein (AFP) gene, construct the eukaryotic expression vector of AFP and express in CHO cells, total RNA were extracted from Hepa 1-6 cells, and then the murine alpha-fetoprotein gene was amplified by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant of vector was identified by restriction enzyme analysis and sequencing. After transient transfection of CHO cells with the vector, Western blotting was used to detect the expression of AFP. It is concluded that the 1.8 kb murine alpha-fetoprotein gene was successfully cloned and its eukaryotic expression vector was successfully constructed.
CHO Cells ; Cloning, Molecular ; Cricetinae ; DNA, Complementary ; Eukaryotic Cells/metabolism ; Genetic Vectors ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; alpha-Fetoproteins/*biosynthesis ; alpha-Fetoproteins/genetics

In order to investigate the relationship between the VEGF level and the counts of dendritic cells (DCs) in peripheral blood of patients with hepatocellular carcinoma (HCC) before and after transcatheter arterial chemoembolization (TACE), the peripheral blood was obtained from 37 patients with HCC who treated by TACE. The blood was obtained on the day before TACE, the first day, the 7th day and the 15th day after TACE respectively. The counts of DCs were quantified by flow cytometry. The plasma VEGF level was measured by ELESA kit. It was shown after TACE, the counts of DCs in peripheral blood were decreased significantly (P<0.05), and the VEGF level in peripheral blood was increased significantly (P<0.05). The counts of DCs in peripheral blood had an inverse correlation with the plasma VEGF level (r=-0.57, P<0.05) after TACE. It was concluded that in patients with HCC after TACE, the increased plasma VEGF level appeared to have the effect to suppress the maturation of DCs, which may contribute to reduction of the body's anti-tumor immunity effect, with a consequence of recur and metastasis of tumor.

Objective To investigate the effect of TGF ? 1 and TGF ? on hepatic apoptosis in hepatocelluler carcinoma (HCC) after transcather arterial chemoembolization (TACE).Methods The specimens were surgically obtained from 36 patients who had been treated by TACE (TACE group) and 19 patients without any treatments (no TACE group). Expression of TGF ? 1 and TGF ? in HCC tissues was detected by immunohistochemical method. Apoptosis was detected by ISEL method.Results Expression rates of TGF ? 1 in HCC tissues of TACE group and no TACE group were 86.11% and 36.84% respectively.expression rates of TGF ? in HCC tissues of TACE group and on TACE group were 0.0088 and 0.1901 respectively.Conclusions After TACE, the expression of TGF ? 1 was enhanced to accelerate the hepatic apoptosis in HCC.TGF ? was mainly related to the cell proliferation in HCC.That the expression of TGF ? was inhibited after TACE might accelerate the hepatic apoptosis of HCC.

Objective To investigate the clinical significance of the expression of fragile histidine triad (FHIT) protein in primary hepatocellular carcinoma (HCC).Methods Immunohistochemical (S-P) method was used to detect the expression of FHIT in the 46 patients with HCC and 10 normal controls.Results In the patients with HCC,the expression rate of FHIT in the tumor tissue was 56.52%,significantly lowerthan that in adjacent non-tumor tissue and normal tissue.The absence of FHIT protein was correlated neither with the size,tumor capsule,serum AFP level nor with chronic hepatitis B virus infection and cirrhosis of liver.There was significant relationship between the expression of FHIT and differentiation and thrombus in the portal vein.Conclusion The loss of FHIT gene protein is a frequent event in HCC.Furthermore,the loss is closoly related to tumor prognosis prognosis and FHIT loss.

Objective To investigate the change of vascular endothelial growth factor (VEGF) expression in HepG2 cells under hypoxia. Methods HepG2 cells were cultured under hypoxia(hypoxia group) and normal condition (control group). VEGF expression of HepG2 cells was examined by immunohistochemical staining. The growth of HepG2 cells was examined by MTT colorimetry and cell count. VEGF level in the culture medium was measured by ELISA.Results After 48 h and 72 h of culture, the growth rate of HepG2 cells in hypoxia group was lower than that in control group (P

Objective To study the expression of survivin and its relationship with the proliferative activity in hepatocellular carcinoma.Methods Using immunohistochemistry ABC methods, the expression of survivin and PCNA were evaluated in 31 hepatocellular carcinoma, 8 cirrhosis of liver and 4 normal livers tissues.Results No expression of survivin were detected in normal livers and cirrhosis of liver. In contrast, survivin protein was expressed in 19 of 31 patients with HCC(61.3%). When compared with normal livers and cirrhosis of liver, hepatocellular carcinoma had significantly more frequencies of survivin expression(P

Objective To observe the relationship between the expression of GFAP and iNOS the post-traumatic intervals after experimental cerebral contusion,in order to attempt to find out a method of timing wound age of brain contusion in the forensic prediction of injury time.Methods After setting up an model of experimental focal cerebral contusion in rats,immunohistochemical method was applied to observing the changes of inducible nitric oxide synthase (iNOS) and glial fibrillary acidic protein (GFAP) staining combined with histology method in rats.The immunostaining results were measured quantitatively with computer imaging analysis system.Results (1) At 3h after injury,GFAP-positive astrocytes around the wounded area were definitely detected.The intensity and the area of GFAP-positive cells increased following the post-traumatic intervals.The intensity peaked dually at 1d and 5d,respectively,followed by reduction at 3d,which revealed two-peak waveforms,The area of GFAP positive cells enhanced significantly at 3h and gradually increased with prolonging survival time,and reached a peak at 7d after injury.(2) At 12h after injury,the iNOS isoform was present and iNOS positive staining cells were noted.The intensity and the area of iNOS positive cells increased following the post-traumatic intervals.The area of positive reached maximal level at 1-3days,gradually subsided after 5d,and maintain a higher lever up to 7d post-injury;but the intensity of individual positive cell gradually increased with prolonging survival time,reached a peak at 5d and gradually subsided after 5d,but the level was still higher than the initial interval.Conclusions Expression of GFAP and iNOS is correlated with post-traumatic intervals after cerebral contusion in rats,suggesting that the expression of iNOS and GFAP may be served as the markers for timing of brain contusion in forensic practice.

Objective To establish a method for quantitative determination of triptolide and triptonide in human plasma samples by HPLC.Methods The samples were extracted with Oasis HLB solid phase extraction cartridge and analyzed with reserved phase HPLC coupled with diode array detection.Results Recoveries rates obtained from spiked plasma for the two diterpenoid were higher than 80%.The linearity ranged from 10 ng/ml to 1 000 ng/ml.Detection limits of the method were 3.0ng/ml for triptolide and 4.5ng/ml for triptonide.Conclusion The method is fast and accurate,which is applicable to determination of diterpenoid in cases of suspected poisoning of Triptergium wilfordii Hook.f. in forensic practice.

Objective To explore how to improve the diagnosis and surgical treatment of primary gallbladder carcinoma. Methods The clinical data of 59 patients with primary gallbladder carcinoma underwent operative treatment in our hospital from January 1985 to December 1999 were retrospectively analyzed. Results There were 22 males and 37 females in this series (M:F=1:1.68). 59%(35/59) patients were complicated by gallstone. Right upper quadrant abdominal pain, jaundice and abdominal mass were the major clinical manifestations. 34 cases underwent cholecystectomy with 57%(34/59) resection rate. Pathological examination showed that gallbladder carcinomas accounted for 94%. Postoperative 1-,3-,5-year survival rates were 36%(21/59), 20%(12/59)and 10%(6/59), respectively. The postoperative survival rate of the early stage patients was higher than that of the advanced patients(P