Objective To observe changes of the ultrastructure and the cell cycle of hepatoma cells exposed to ultrasound-mediated disruption. Methods Suspensions of Walker-256 hepatoma in vitro exposed to ultrasound disruption with the acoustic power output 0 22 W and the ultrasound frequency 42 kHz were observed by the scanning electron microscopy, the transmission electron microscopy and the flow cytometric analysis of DNA content. Results Cells after ultrasound disruption revealed by scanning electron microscopy exhibited ruptured membrance, disordered organellae and disrupted nucleus, showing typical severe destruction of the cells. Flow cytometric analysis of DNA content demonstrated an increased in the population of cells in the G 2 and M phases of the cell cycle but a decrease in number of cells in the S phase, suggesting an inhibition of the cell synthese and an increased sensitivity to radiation. Conclusions Ultrasound disruption can destroy tumor cells effectively at the cellular level.

Objective To identify clinicopathologic factors which predict survival following hepatectomy in HBV-related cirrhotic patients with early hepatocellular carcinoma (HCC).Methods A database was used to identify patients with histologically confirmed early HCC (≤5 cm,no nodal involvement,metastases,or major vascular invasion) who underwent surgical resection (excluding ablation or transplantation).Among 20 700 patients with HCC who were diagnosed at the Eastern Hepatobiliary Surgery Hospital from April 2005 to November 2010,537 (2.6％) patients with early HCC were studied retrospectively.Prognostic factors were evaluated using the Kaplan-Meier curves,Cox proportional hazards models and the receiver operating characteristic (ROC) curves.Results The study included 537 patients.The median tumor size was 2.9 cm,and 33％ of patients had tumors ≤2 cm.Most HCC lesions were solitary (63％) and had no evidence of vascular invasion (64％).Following surgery,the overall median and 5-year survival were 45 months and 33％ respectively.After adjusting for demographic factors and histological grade,tumor size ＞2 cm (hazard ratio [HR]:1.56),multifocal tumors (HR:1.34),and vascular invasion (HR:2.03) remained independent predictors of poor survival (all P ＜ 0.05).Based on these findings,a prognostic scoring system was developed that allotted 1 point each for these factors.Patients with early HCC could be stratified into 4 distinct prognostic groups (median and 5-year survival,respectively):0 points (97 months,96％),1 point (85 months,76％),2 points (76 months,54％),3 points (56 months,39％) (P ＜0.01).Conclusions The present study emphasized the importance of pathologic staging even in patients with small HCC.Anatomical resection of HCC should be the preferred surgical procedure in cirrhotic patients.

Objective To study the mechanism of cytotoxic T lympocytes in the acute rejection of xenogenic liver transplantation(XLT). Methods Hamster to rat orthotopic liver transplantation model was performed with three cuff method. In XLT,lymphocytes including CD8 and CD4 subsets were observed by histology and immunohistochemistry; the expression of perforin and Fas- L was observed by immunohistochemistry; and apoptotic cells of XLT were observed in situ end-labelling of fragmented DNA. Results In the XLT, T cell infiltration firstly ocurred on the 2nd day posttansplantation and located at the portal triads; the infiltrating lymphocytes proliferated increased with time and reached the peak on the 5th to 6th days.Perforin and Fas-ligand were expressed on the 4th day after XLT, and peaked on the 5th to 6th day,and the expression of perforing was still higher than the expression of Fas-ligand. Conclusions T lymphocyte participates in the acute rejection of XLT through the expression of perforin and Fas-ligand, which leads to apoptosis or necrosis of hepatocytes.

Objective: To investigate the cytotoxicity of ICE gene transfection in Combination with Antitumor Chemicals killing Hepatocellular Carcinoma Cells in vitro.Methods: The recombinant plasmid pLXSN-hICE was transferred to virus packing cell PA317 by electroporation method. And then the retrovirus containing human ICE cDNA generated by these PA317 cells were used to transfect human hepatocellular carcinoma cell line HepG2. The apoptosis of transferred cells were examined by gel electrophoresis. The influence of chemotherapeutic drug Carbo-platin to the proliferation of hepatocellular carcinoma cell line SMMC7721 and its derivative cells(SMMC7721-hICE,SMMC7721-antisence hICE,SMMC7721-neo)was observed by incorporation of 3H-TDR. Results: Electrophoresis of DNA displayed the apoptosis ladder of HepG2 transfected by ICE gene. The proliferation of SMMC7721-hICE was significantly suppressed in vitro induced by Carbo-alatin compared to the other three cell lines. Conclusion: ICE gene transfection could greatly increase the susceptibility of SMMC7721 cells to apoptotic cell death following chemotherapy. These findings suggest that combining ICE gene transfection with utilizing antitumor drugs would represent a novel approach for the effective treatment of hepatocellular carcinoma.

To study the significance of the expression of splice variants of CD44 (CD44v) in tu-m0r-adjacent tissue of the patients with primary liver cancer(PLC). Methods: To research into the signifi-cance of the expression of CD44v mRNA in tumor-adjacent tissue of 30 patients with PLC by RT-PCR andfollow-up. Results: In the patients that the expression of CD44v mRNA of tumor-adjacent tissue washigher than those of tumor tissue(group I ), clinical pathological indexes were higher than in the patientsthat the expression of CD44v mRNA was higher than the tumor tissue(group I ). The recurrent rate ofgroup I was higher than that of group n (P

To determine the content of dendrobine in Dendrobium nobile from different harvest times and plant parts, to research the inherent rule about it. GC with internal standard was used to determine. The content of dendrobine had significant differences in different periods and parts. The dendrobine content is higher in four-year root than in three-year root. The dendrobine content in the upper segment of stem is the highest, secondly is in the middle seg-ment, and in the low segment is the lowest. This offered evidence to determine the most appropriate harvest time and fair use of different parts for D.nobil.

Objective To study the protection against hepatic ischemia-reperfusion injury by human IL-10 gene transduction in rats. Methods Ad-hIL10-EGFP (1. 0 ? 109 plaque forming units/ml) was administered into SD rats by intravenous injection 72 hours before hepatic ischemia-reperfusion injury was induced. Liver function were tested and HE pathology was observed. The expression of hIL-10 was studied with ELISA or immunohistochemical method, the expression of EGFP was observed in frozen sections under the fluoroscopy. The apoptosis of hepatocytes was observed with Tunel's assay. Results Compared with control rats, the expression of EGFP and hIL-10 was observed, serum hIL-10 level was (815.74 ? 284. 76) ng/ml, liver function of treatment rats were improved, the paraffin sections showed that the hepatocytes were not significantly swelling and liver pathology ameliorated, the number of apoptosis cells decreased (P

Objective To construct a new replicating adenovirus vector CNHK600-p53 carrying anti-tumor gene p53 and investigate its effect on hepatocarcinoma cell line PLC/PRF5. Methods The methylthiazolyl tetrazolium assay (MTT) was used to observe the killing effect of Fluorouracil, Mitomycin and Epirubicin alone or in combination with adenovirus CNHK600-p53 on PLC/PRF5. Results When 5-Fu at concentration of 400?g/ml, the inhibition rate was (65?4. 2) % ; with MMC at 1?g/ml, the rate was (41?1.9)%; and it was (65?1.8)% when EPI at concentration of 10?g/ml. With PLC/PRF5 infected by adenovirus CNHK600-p53 ( MOI = 0. 625) , the inhibition rate was significantly increased to (89?5. 3)%,(60?2.3)% and (75?1.5)% respectively; When MOI was 0.3125, 0.625, 1.25, the inhibition rate in CNHK600-p53 group and Ad-p53 group was (27?2. 5)% , (30?3. 7)% , (61?4. 3)% and(4?2.7)%, (5?3.5)%, (16?4.5)% respectively. Conclusion For hepatocarcinoma cell line PLC/PRF5 the effect of CNHK600-p53 is stronger than Ad-p53.

0.05). The mean growth values in AdCMV-tk/GCV- and AdVEGF-tk/GCV-treated tumors were significantly lower than those in untreated tumors and AdVEGF-tk/saline-untreated tumors( P

Objective To study the effects of CTLA4-Ig on mu rine allergic contact dermatitis.Methods Mice were exposed to DNFB to induce allergic contact dermatitis and were i njected with CTLA4-Ig.Ear swelling was measured 24h after antigen challenge.Splenocytes from treated mice were assayed for their ability to prolif erate in response to DNFB or FITC stimulation in vitro.Results Profound inhibition of contact hypersensitivity response(CHS )was shown by 69.7%in mice treated with CTLA4-Ig compared with mice treate d with PBS control.CT-LA4-Ig-treated mice displayed DNFB-specific tolerance,but exhibited a vigorous immune response to FITC when re-sensitizing 14days after the fir st challenge.Adoptive transfer of l ymphocytes from CTLA4-Ig-treated mice could induce inhibition of CHS in recipien t mice.Conclusions CTLA4-Ig can inhibit CHS by blocking B7/CD28co-stimulatory pathway,which provides a new way to suppress typeⅣallergic reaction.[