Cholangiocarcinoma is a malignancy of biliary tract and its incidence has been increasing in recent years all over the world.Meanwhile,there have been advances in techniques for its diagnosis and treatment.Some new methods such as PET,endoscopic ultrasonography and scopic CT can promote the diagnostic rate in the early stage and they have been used for diagnosis and staging of tumors.For those patients with large cholangiocarcinoma in the liver,photodynamic therapy is an effective neoadjuvant treatment.For those patients with unresectable cholangiocarcinoma,organ transplantation might be employed. Classification of cholangiocarcinoma and study on its diagnosis are important for disease evaluation in patients with the disease.

ObjectiveTo study the effect of anti-oncogene p15 and p16 on the proliferation of human cholangiocarcinoma cell line.MethodsThe cDNA of anti-oncogene p15 and p16 was constructed into pcDNA3-neo plasmid carrier. The human cholangiocarcinoma cell line QBC939 were transfected with the recombinants pcDNA3p15 and pcDNA3p16 using lipofectin, respectively. The expression products were analyzed by Western blot. Cell viability and death were measured with MTT assay. Cell cycle was determined by flow cytophotometry and the formation of cell clone was detected. Results The growth of QBC939 cells was inhibited. The flow cytophotometry verified p15 and p16 induced QBC939 cell G1 blockade. Conclusion Anti-oncogene p15 and p16 together lead to the inhibition of cell cycle.

Objective To investigate the features of immune response of human immunodeficiency virus type-1 (HIV-1) antigen specific T lymphocytes in HIV-1 monoinfected or HIV 1/hepatitis C virus (HCV) coinfected individuals.Methods Twenty-six HIV-1 monoinfected and 23 HIV-1/HCV coinfected individuals were enrolled.Immunomagnetic microbeads were used to isolate T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from human peripheral blood mononuclear cells (PBMC).Frequencies of interferon-γ (IFN-γ) secreting cells of CD4+,CD8+ T lymphocytes and PBMC stimulated by a peptide pool containing 12 overlapping peptides in HIV-1 P24 from 49 patients were assessed by enzyme-linked immunospot (ELISPOT) assay.HIV-1 RNA levels of these patients were also detected by real-time fluorescence quantitative polymerase chain reaction.The data were compared by one-way ANOVA and Mann-Whitney U test,and Spearman test was used for correlation analysis.Results Frequencies of HIV-1 antigen specific CD4+ T lymphocytes [median =25 spot-forming cells (SFC)/106 cells] were significantly lower than those of CD8+T lymphocytes (median=38SFC/106 cells,F=4.592,P=0.037) and PBMC (median=53 SFC/106 cells,F=5.436,P=0.025) in HIV-1 monoinfected group.Frequencies of HIV-1 antigen specific CD4+ T lymphocytes (median=5 SFC/106 cells,Z=-2.432,P=0.015),CD8+T lymphocytes (median=5 SFC/106 cells,Z=-1.996,P=0.046) and PBMC (median=10 SFC/106 cells,Z=-2.306,P=0.021) in HIV-1/HCV coinfected group were significantly lower than those in HIV-1 monoinfected group.Conclusions In HIV-1 infection,antigen specific immune response of CD4+ T cells can be activated,but weaker than that of CD8+ T cells.Co-infection with HCV might down-regulate the responses of HIV-1 antigen specific T lymphocytes in HIV-1 infected individuals.

Objective To evaluate the effect and mechanism of non-steroidal anti-flammatory drags(NSAIDs) on the colon cancer cell growth by using S-nitrosoglutathione(GSNO) which can produce nitric oxide.(Methods) Apoptosis of 3 colon cell lines were evaluated by cell growth curve and flow cytometry,the PGE_2 levels in cell culture supernatants were determined by competitive enzyme immunoassay method,and the(protein) expression of COX-1 and COX-2 were analyzed by Western blot.Results The production level of PGE_2 was increased with CSNO treated concentration and time.Using 500?mol/L CSNO treatment for 48h,the expression level of COX-1 and COX-2 protein increased.NASIDs can block the production of PGE_2 but had no effect on the inhibition of cell growth induced by GSNO.Conclusions GSNO can increase PGE_2(production) and induce COX-1 and COX-2 protein expression in a dose-and time-dependent manner.Higher concentrations of GSNO also can inhibite cell growth and induced apoptosis in all 3 cell lines.NSAIDs can block production of PEG2 but NSAIDs are no effect on cell growth.

ObjectiveTo explore the feasibility, safety and clinical value of laparoscopic Rouxen-Y cystojejunostomy in the treatment of pancreatic pseudocyst. Method Four patients with pancreatic pseudocyst received totally laparoscopic pancreatic pseudocystojejunostomy. The data on intraoperative bleeding, operative time, postoperative time to get out of bed, time of first flatus/bowel motion, complication and duration of hospital stay were collected and analyzed retrospectively. ResultsAll operations were carried out successfully with laparoscopic surgery. The mean operative time was 90 min. The average intraoperative blood loss was 40 ml. The mean postoperative time to get out of bed was 1.5 d, and the mean time of first flatus/bowel motion was 2. 3 d. All patients recovered smoothly without any pancreatic fistula. The average hospital stay was 7 days. Fever, pancreatitis,adhesive intestinal obstruction and other complications did not occur. ConclusionsTotally laparoscopic Roux-en-Y pancreatic pseudocystojejunostomy was an efficacious, safe, and minimally invasive procedure.

Objective To investigate the expression of pSTAT5 in 7 pancreatic carcinoma cell lines,and the change of expression of pSTAT5 in pancreatic carcinoma cells SW1990 after growth hormone (GH) treatment, and explore its molecular mechanism. Methods Human pancreatic carcinoma cell lines (SW1990, Cap-1, Colo, Mia, AsPc, P3, PANC1) were cultured in vitro, and Western blotting was used to detect the expression of pSTAT5 in these cell lines. SW1990 in exponential growth phase was collected and nude Balb/c mice were inoculated with SW1990 cells. When tumors became palpable after inoculation, mice (normal saline group). 1 h, 2 h and 24 h after the last dose of GH treatment, the mice were sacrificed.Western blotting was used to detect the expression of pSTAT5 in SW1990 and inoculation tumor cells after GH injection. Results Positive expression of pSTAT5 was observed in all human pancreatic carcinoma cell lines (SW1990, Cap-1, Colo, Mia, Aspc, P3, PANC1). 5 minutes after GH (50 ng/ml) stimulation, the expression of pSTAT5 in SW1990 was 0.57 ±0.05, which was significantly increased; and it reached 0.64 ±0.04 at 10 minutes, then decreased to 0.39 ±0.03 at 15 minutes, however, it remained higher than that in the control group at 1 h (0.33 ± 0.02 vs 0.25 ± 0.06), and its expression at 2 h was 0.26 ± 0.03 and returned to the normal level. The expression of pSTAT5 in xenograft was not significantly changed. Conclusions GH could rapidly up-regulate the expression of pSTAT5 in SW1990 but the effect lasted for a relatively short period. GH had no significant effect on the expression of pSTAT5 in xenograft.

Objective To investigate the effect of GH on proliferation of pancreatic cancer cells and observe the features of IGF-IGFBP3 pathway in the host after GH administration. Methods Pancreatic cancer cells (SW-1990,PANC-1 and P3) during exponential growth stage were harvested and cultured in medium containing growth hormone (50 ng/ml). After 24, 48 and 72 hours, cells were counted using a Coulter Counter. Thirty-five Athymic nude Balb/c mice were inoculated with SW-1990cells. When tumors became palpable after inoculation, animals were randomized to receive GH points (1 h, 2 h, 6 h, 24 after the last injection), plasma samples were gathered for subsequent ELISA determination and liver was rapidly incised for immune blotting analysis. Results The results revealed that GH stimulated cell growth in vitro. GH elevated levels of IGF-Ⅰ , Ⅱ at the 1st , 2nd , 6th hour after the last injection. GH augmented the expression of IGFBP3 in the liver of the host in vivo (1 h, 2 h, 6 h, 24 h, respectively). Conclusion Such proteins as IGF- Ⅰ and Ⅱ might be associated with mechanism of last effect of GH on tumor host. The up-regulation of IGFBP3 by GH administration in the host may help to explain the phenomena that GH doesn't accelerate growth of pancreatic tumor in vivo.

Objective To study the impact of exogenous growth hormone (GH) on the levels of insulin-like growth factor-Ⅰ and -Ⅱ (IGF-Ⅰ, -Ⅱ) of the pancreatic cancer tissue and the small intestine mucosa of the host. Methods In situ hybridization was performed on pancreatic cancer cell lines (SW-1990) and inoculation tumor of the host to determine the location of the mRNA transcript encoding IGF R-Ⅰ,-Ⅱ. Athymic nude Balb/c mice were inoculated with SW-1990 cells. After inoculated tumors have become palpable, animals were randomized to receive GH (4 mg/kg once daily for 2 weeks) versus saline control. After the animals were killed at time point, tissues (tumor and small intestine) were rapidly incised for subsequent immune blotting analysis. Results Strong IGF R-Ⅰ,-Ⅱ mRNA hybridization signal could be detected in pancreatic cancer cell. There was no statistically significant difference between the level of IGF-Ⅰ, Ⅱ in the tumor of the GH and NS groups after 1 hours of GH injection (P>0.05). GH augmented the expression of IGF-Ⅰ(1 h : 0.33±0.05, P<0.05 ; 2 h : 0.34±0.04, P<0.05 ; 6 h:0.34±0.05, P<0.05), -Ⅱ(1 h : 0.36±0.05, P<0.05) in the small intestine mucosa of the host. Conclusions The expression of IGF-Ⅰ, Ⅱ in the small intestine mucosa of the host was elevated by GH, but not in the inoculation tumor in vivo. The discrepancy of GH-IGF pathway between inoculation tumor and small intestine of the host may help to explain the phenomena that GH doesn't accelerate growth of pancreatic tumor in vivo.

To explore the effects of Dachengqi Decoction (DCQD), a compound traditional Chinese herbal medicine, on pancreatic acinar cell apoptosis in a rat model of experimental acute pancreatitis.

Objective To investigate the effect of highly active antiretroviral therapy (HAART) on human immunodeficiency virus type-1 ( HIV-1 ) antigen specific cytotoxic T lymphocyte (CTL) immune responses. Methods Peripheral blood mononuclear cells (PBMCs) were collected from 38 HIV-1 infected individuals receiving HAART ( HAART group) and 31 HIV-1 infected individuals not receiving HAART (non-HAART group), and stimulated with a peptide pool containing 12 overlapping peptides in HIV-1 P24;then the frequency of interferon γ ( IFNγ ) secreting cells were assessed by enzyme-linked immunospot (ELISPOT) method. Difference in HIV-1 antigen specific CTL immune response between non-HAART group and HAART group was analyzed by χ2 and Mann-Whitney U tests. Results Positive response rate of HIV-1 antigen specific CTL immune responses in HAART group ( 65.8%, 24/38 ) was higher than that of non-HAART group (32.3%, 10/31, χ2 = 6. 522, P ＜ 0.05 ). For HIV-1 infected individuals with blood CD4 +T cells ＞ 350/μL, the frequency of HIV-1 antigen specific CTL responses in HAART group was higher than that in non-HAART group (Z = -2. 819, P ＜0.05 ). In the HAART group, those receiving HAART more than 12 months were of higher frequency of HIV-1 antigen specific CTL responses ( Z =-2. 195, P ＜ 0. 05 ). Conclusion HAART especially long-term treatment may enhance HIV-1 specific CTL responses in HIV-1 infected individuals.