Objective To assess clinicopathological features and prognosis of patients with combined hepatocellular and cholangiocarcinoma (cHCC-CC). Methods Clinicopathological and follow-up data of 115 cHCC-CC patients confirmed pathologically in Liver Cancer Institute of Fudan University from 1995 to 2007 were analyzed. Kaplan-Meier method was used to calculate 1-,3- and 5-year survival rates and tumor-free survival rates. Survival curves were analyzed using the log-rank test. The factors that impacted the prognosis of cHCC-CC were estimated. Results In 115 cases, one was Allen's type A, one was Allen's type B, and the other 113 were Allen's type C. Being with male in predominance, most of the cHCC-CC patients had liver cirrhosis background. They presented with elevated AFP or CA19-9, vascular invasion, resembling hepatocellular carcinoma(HCC)as well as lymph nodes metastasis. One-, 3-, 5-year survival rates of 115 patients were 68. 1%, 38. 1% and 33.6%, respectively, with median survival time of 13.0 months. Whereas the 1-, 3-, 5-year survival rates in radical resected patients were 78.4 % ,44.4 % and 44.4 % ,respectively, with median survival time of 16.0 months. Tumor free survival time at 1-, 3- and 5-year was 57.8 %, 12.6 % and 0.0 %,respectively,with median recurrent time of 10.0 months. One-, 3-, 5-year survival rates of 10 nonsurgical patients were 10/10,10/10 and 0/10,respectively, with median survival time of 5.3 months.TNM stage was independent factor for prognosis of the patients after resection. Whereas the lymph nodes involvement was independent factor for the tumor free survival time of radical resected patients.Conclusions Although clinicopathological characteristics of cHCC-CC are more similar to those of HCC, the prognosis of cHCC-CC is more unfavorable than that of HCC. TNM stage is an independent determinant of long time outcome for patients after resection.

Objective To evaluate the value of MSCTA in combination with CTV in diagnosis of pulmonary embolism and deep venous thrombosis (DVT) of lower limb. Methods 47 patients with pulmonary embolism proved clinically were examined with MSCTA using 120 kV,210 mA,collimater of 0.75 mm and the automatic bolus traching technique,the speed of contrast agent infu-sion was 3～4 m/s after 170 s delayed,venous scan of lower limbs was performed. Results 47 cases were well perform the trunk of bilateral pulmonary artery, pulmonary lobar artery, pulmonary segemental artery, pulmonary inferior segmental artery could be showed by MSCTA, MPR, MIP and VRT. 363 arterial segments were involved and 31 cases accompanied with DVT. Conclusion MSCTA in combination with CTV is of significant value in detecting pulmonary embolism accompanied with deep venous thrombo-sis.

Objective To explore the values of CT virtual gastroscopy (CTVG), three dimensional (3D) and axial imaging by multi slice spiral CT (MSCT) in the preoperative assessment of gastric carcinoma. Methods Fifty two patients with gastric carcinoma proven by means of biopsy underwent CTVG, 3D imaging using the surface shaded display (SSD) and Raysum, and axial imaging. MSCT was first performed after the ingestion of gas, and a second tri phase MSCT scan was performed after the ingestion of water. The results were compared with those of fibrous gastroscopy (FG) and surgical pathological findings. Results The detection rates of gastric lesions with CTVG, 3D and axial imaging by MSCT were 98.1%, 96.2%, and 96.2%, respectively. The accuracy of judging gross type of advanced gastric carcinoma using CTVG combined with 3D imaging, FG,and axial imaging was 92.5% , 82.5%, and 75.0%, respectively.Statistical difference existed in CTVG combined with 3D imaging and axial imaging (P

Objective To investigate the selective oncolytic role and antitumor action of a novel recombinant adenovirus containing E1A and IL-24 on hepatocellular carcinoma cell(HCC). Methods The recombinant adenovirus expressing IL-24 (Ad. HS4. AFP. E1A/IL-24) was constructed by using modified human alpha-fetoprotein (HS4-AFP) promoter to drive adenovirus E1A gene and II-24 gene.Cell Counting Kit-8 were performed to test the selective cytotoxicity of the virus in hepatocellular carcinoma cell lines SMMC-7721, Hep3B, MHCC97-H and hepatocyte cell line L02 . The mRNA and protein expression of IL-24 gene were detected by RT-PCR and western blot. Cell growth curves and Annexin V/PI assay were used to study cell proliferation and apoptosis of MHCC97-H. The anti-metastatic effects of the recombinant adenovirus were evaluated in cell adhesion, migration, and cell motion. Matrix metalloproteinase-2 (MMP-2) expression was examined by RT-PCR and zymography.Results Selective replications of Ad. HS4. AFP. E1A/IL-24 adenovirus were observed in over expression AFP cell line MHCC97-H, a highly metastatic potential HCC cell line but not in hepatocyte cell line L02. The mRNA and protein of IL-24 were also over expressed in MHCC97-H. This recombinant adenovirus also showed the significant oncolytic action on MHCC97-H but not on L02 (P＜0. 05). Besides, the recombinant adenovirus significantly inhibited MHCC97-H metastatic potential such as cell adhesion, migration and invasion as well(P＜0.01). Conclusion The selective oncolytic adenovirus expressing E1A and II-24 has a selective antitumor effect and play an inhibitory role in metastasis of HCC.

Objective:To observe the change of periphery and centra lymphocyte subsets at the crest-time of MOG_(35-55) induced EAE disease in mice,and to explore the alteration of cellular immunity and humoral immunity in the invasion process in EAE.Methods:MOG_(35-55) was used to establish EAE model in femina C57BL/6 mice.The behavioral changes and the histological scores were recorded after the mice were immuned .The changes of CD3~+CD4~+,CD3~+CD8~+,CD4~+CD25~+ and B220~+ on periphery and centra lymphocytes in spleen,brain and spinal cord were analyzed by flow cytometry.Results:The CD3~+CD4~+,CD3~+CD8~+,CD4~+CD25~+ and B220~+ lymphocytes were detected in the brain and spinal cord of EAE group mice,but they were not detected in CFA control group.The CD3~+CD4~+ and CD3+CD8+lymphocytes in the spleen of EAE crest-time group were lower than those in CFA control group(P＜0.05).The B220~+ lymphocytes were obviously higher than in the CFA control group (P＜0.01).And CD4~+CD25~+ lymphocytes were slight higher than the CFA control group.Conclusion:At the crest-time during EAE,the CD3~+CD4~+,CD3~+CD8~+lymphocytes of spleen reduced obviously,B220~+ lymphocytes increased markedly,and the CD4~+CD25~+ lymphocytes just have the increasing trend.It indicates that cellular immunity and humoral immunity coregulated the patho-process at the crest-time of EAE,T lymphocytes and B lymphocytes all played important roles in the pathogenesy of EAE.

A group of SARS-Iike coronaviruses(SL-CoV)have been identified in horseshoe bats.Despite SL-CoVs and SARS-CoV share identical genome structure and high-level sequence similarity,SL-CoV does not bind to the same cellular receptor as for SARS-CoV and the N-terminus of the S proteins only share 64% amino acid identity,suggesting there are fundamental differences between these two groups of coronaviruses.To gain insight into the basis of this difference,we established a recombinant adenovirus system expressing the S protein from SL-CoV(rAd-Rp3-S)to investigate its immune characterization.Our results showed that immunized mice generated strong humoral immune responses against the SL-CoV S protein.Moreover,a strong cellular immune response demonstrated by elevated IFN-γ and IL-6 levels was also observed in these mice.However,the induced antibody from these mice had weaker cross-reaction with the SARS-CoV S protein,and did not neutralize HIV pseudotyped with SARS-CoV S protein.These results demonstrated that the immunogenicity of the SL-CoV S protein is distinct from that of SARS-CoV,which may cause the immunological differences between human SARS-CoV and bat SL-CoV.Furthermore,the recombinant virus could serve as a potential vaccine candidate against bat SL-CoV infection.

Objective To explore the clinical characteristics and surgical treatment of gastric cancer in elderly patients.Methods The clinical data of 431 patients older than 70 years with gastric cancer from 1984 to 1998 were analyzed retrospectively.Results The main clinical manifestations were upper abdominal pain,distension or upset,loss of appetite and weight.The patients with clinicalⅢ,Ⅳ stage accounted for 64.2 percent.Pathological findings showed no specificity.Radical gastrectomy was performed in 215 cases(49.9%),and palliative resection was done in 122 ones(28.3%).The operative mortality and postoperative complication rate were 5.1%and 25.5%in the patients respectively.The 5-year survival rate was 29.5%in the patients with surgery,53.2%with radical gastrectomy and 10.7%with palliative resection.There were significant differences in 5-year survival rate between the radical and palliative groups.Conclusions The majority of elderly patients are clinical Ⅲ,Ⅳ stages and their operative complication rate is high.The key to increase the survival rate and life quality of elderly patients with gastric cancer is early diagnoses,strong perioperative management and suitable radical surgery.

Pig copper zinc superoxide dismutase (CuZnSOD) is an important antioxidant enzyme. Some studies focused on the function of CuZnSOD gene, but the transcriptional regulation of the CuZnSOD gene is not yet fully elucidated. Therefore, the aims of the study were to determine the core promoter region and to explore its mechanism of transcriptional regulation. The 853 bp DNA sequence of 5'-flanking promoter was amplified by performing PCR. A series of CuZnSOD promoter fragments with gradually truncated 5'-end were produced by nested PCR and inserted into pGL3-Basic vector. The activities of the promoters were measured by the dual-luciferase assay system after transient transfection into the NIH/3T3 cells. The results demonstrated that there were 2 potential transcription start sites in the regions from initiation codon to -87 bp and -266 bp, respectively. The region from -383 bp to +67 bp in CuZnSOD gene promoter showed higher activity than other regions, and further deletion analysis demonstrated that the region from -75 bp to -32 bp contained an essential promoter sequence for pig CuZnSOD gene transcription. In addition, several potential transcription factor binding sites were predicted with bioinformatics method. These results suggest that these transcription factor binding sites may be involved in the transcriptional regulation of CuZnSOD gene.
Animals ; Cloning, Molecular ; Gene Expression Regulation ; Genetic Vectors ; Mice ; NIH 3T3 Cells ; Promoter Regions, Genetic ; Superoxide Dismutase ; genetics ; Swine ; Transfection

Objective To investigate the effect of unfractionated heparin (UFH) on the expression of heme oxygenase-1 (HO-1) in intestinal mucosa of mice with sepsis.Methods Thirty-six male C57BL/6J mice were randomly divided into sham group,cecal ligation and puncture (CLP) group and UHF group,n =12 in each group.Model of intestinal injury in sepsis was induced by CLP.In sham group,the mice were exposed without ligation of cecum.In UFH group,the mice were treated intravenously with 8 U of UFH via the tail vein half an hour before the operation and 12 hours after the surgery respectively.Six mice in each group were randomly chosen at 4 hours and 24 hours after operation to collect inferior vena venous blood samples and terminalileum tissues.The serum levels of interleukins (IL-1 β,IL-6),and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA).The serum level of D-lactate was determined by colorimetry.Pathological changes of ileum tissue and Chiu score were observed after hematoxylin eosin (HE) staining.The HO-1 expression was detected immunohistochemically.Results In sham group,no significant changes in the serum levels of IL-1 β,IL-6,TNF-α and D-lactate were observed.Twenty-four hours after the operation,the structure of intestinal mucosa was basically normal without obvious pathology change and no HO-1 positive cells were found.The serum levels of IL-1 β,IL-6,TNF-α,and D-lactate in CLP group were gradually increased,and they were significantly increased as compared with sham group [IL-1 β (ng/L):40.87±2.88 vs.22.60±2.05 at 4 hours,113.73±3.96 vs.22.07±2.74 at 24 hours;IL-6 (ng/L):63.89±3.26 vs.44.89±3.38 at 4 hours,176.56±5.45 vs.45.76±4.02 at 24 hours;TNF-α (ng/L):194.62± 14.13 vs.152.05±6.22 at 4 hours,599.62± 10.20 vs.155.90± 14.18 at 24 hours;D-lactate (mmol/L):0.24± 0.02 vs.0.19 ± 0.01 at 4 hours,0.33 ± 0.04 vs.0.20 ± 0.02 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,edema and inflammation in ileal mucosa,intestinal villi structural damage were observed,the Chiu score was significantly higher than those in the sham group [4.5 (3.0-5.0) vs.0 (0-1.0),P ＜ 0.05],and a small amount of HO-1 positive cells were localized in the intestinal mucosa.Compared with CLP group,the serum levels of IL-1 β,IL-6,TNF-α,and D-lactate of UFH group were significantly decreased [IL-1 β (ng/L):31.53 ± 2.90 vs.40.87 ± 2.88 at 4 hours,61.13 ± 2.80 vs.113.73 ± 3.96 at 24 hours;IL-6 (ng/L):51.16 ± 5.68 vs.63.89 ± 3.26 at 4 hours,81.16 ± 4.54 vs.176.56 ± 5.45 at 24 hours;TNF-α (ng/L):171.76± 5.60 vs.194.62± 14.13 at 4 hours,328.48 ± 10.79 vs.599.62± 10.20 at 24 hours;D-lactate (mmol/L):0.21 ±0.01 vs.0.24±0.02 at 4 hours,0.24±0.02 vs.0.33±0.04 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,intestinal injury was ameliorated,the Chiu score was significantly lower [1.5 (1.0-5.0) vs.4.5 (3.0-5.0),P ＜ 0.05],and HO-1 positive cells in the intestinal mucosa was remarkably increased.Conclusion UFH can enhance the expression of HO-1 in intestinal mucosa,reduce the release of inflammatory factors,ameliorate the intestinal inflammatory response,and thus play a protective role in intestinal tissue in mice with sepsis.

BACKGROUND:Unlike the ilium derived from the paraxial mesoderm, the mandible from cranial neural crest has a unique mechanism. Core binding factorα1 (Cbfα1) is a key transcription factor for skeletogenic process. However, the role of Cbfα1/p56 subtype in mandible tissue is yet not clear. OBJECTIVE:To research the expression of Cbfα1/p56 subtype in bone marrow mesenchymal cells from rat mandible in vitro. METHODS:Bone marrow mesenchymal stem cells from rat mandible and ilium were in vitro isolated and purified by primary culture. The characteristics of bone marrow mesenchymal cells were compared through the methods of enzyme linked immunosorbent assay and real-time PCR, including growth curve, alkaline phosphatase activity and relative mRNA expression of Cbfα1 subtypes. RESULTS AND CONCLUSION:Bone marrow mesenchymal cells from rat mandible and ilium were successful y obtained. Bone marrow mesenchymal cells from the mandible proliferated more rapidly, alkaline phosphatase activity of which was higher than iliac cells. The relative mRNA expression of Cbfα1/p56 subtype in bone marrow mesenchymal cells from the mandible was more than that in iliac cells at 6 days of culture (P<0.05), while the expression of Cbfα1/p57 in each time showed no statistical significance (P>0.05). The results showed that Cbfα1/p56 is very significant in the early osteogenic differentiation of bone marrow mesenchymal cells from the mandible.