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Peutz-Jeghers syndrome [PJS] is characterized by scattered black pigmentations on fingers and lips and multiple polyps in the gastrointestinal tract. Patients with PJS often have severe complications secondary to multiple large polyps. Herein, we present a young woman complicated with a large polyp in her colon without any clinical symptoms. We also emphasized the necessity of early diagnosis and treatment of multiple polyps in such patients
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Objective To investigate the ability of induction of specific cytotoxic T lymphocytes (CTL) stimulated by dendritic cells (DCs) co-transfected with MUC1 and survivin mRNA of human pancreatic cancer,and to provide the experimental basis for the treatment of human pancreatic cancer with multi-epitope DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) of 6 patients with pancreatic cancer.Human pancreatic cancer cell line MiaPaCa-2 was routinely cultured,after being transcripted and amplified by RT-PCR,MUC1 and survivin mRNA were co-transfected or individually transfected into DCs by electroporation,and they were named as DC-MUC1,DC-survivin,DC-MUC1 + survivin.The expression of MUC1 and survivin mRNA in DCs were detected by real-time PCR.The survival rate of transfected DCs were determined by MTT method.The lymphocyte proliferation ability was evaluated by mixed cell culture method.The Th1 cytokine releasing of antigen-specific CTLs were measured by ELISA assay.Results Mature DCs were obtained,the positive expression rates of surface markers CD40,HLA-DR,CD83 and CD86 were 34.31％,50.21％,89.17％ and 73.62％,respectively.The expression amount of MUC1 mRNA of DC-MUC1 was 36.24 ± 5.17,and the expression amount of survivin mRNA of DC-survivin was 34.53 ± 4.02,while the expression amounts of MUC1,survivin mRNA of DC-MUC1 + surviving were 31.79 ±4.26 and 14.67 ± 2.96,which were significantly lower than that in individual transfection group (P ＜ 0.05).The survival rate of DC-MUC1 + surviving was decreased in a time dependent manner,which was significantly lower than that in individual transfection group (about 50.21％ vs 80％ at 24 h,P ＜0.05).When DC/T cells ratio was 1∶ 10,1∶ 20,the autologous T cell proliferation index of MUC1 and survivin mRNA in co-transfection DC group was significantly higher than that in individual transfection group (P ＜ 0.05) ;when DC/T cells ratio was 1∶ 40,1∶ 80,the difference of proliferation index was not statistically significant.When DC/T cells ratio was 1∶ 10,after 14 d culture,the expressions of IL-2 in DC-MUC1,DC-survivin,DC-MUC1 + surviving were (892.73 ± 32.9),(713.62 ± 56.37),(1884.37 ± 95.21) pg/ml,and the expressions of granzyme B were (501.62 ± 12.30),(203.84 ± 12.55),(1193.15 ± 86.04) pg/ml ; and the expressions of IFN-γ were (981.50 ± 47.82),(696.05 ± 41.66),(2237.94 ± 189.55) pg/mL.The corresponding values in DC-MUC1 + surviving group were significantly higher than those in individual transfection group (P ＜ 0.05) ; while the difference of IL-10 was not statistically significant.Conclusions DCs co-transfected with MUC1 and survivin mRNA have a stronger ability to stimulate specific CTL in vitro than individual antigen loaded DCs.
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Objective To investigate the induction of specific anti-tumor immune response by transfected dendritic cells (DCs) with MUC1 mRNA of human pancreatic cancer,and to provide the experimental evidences for the treatment of human pancreatic cancer with DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs),and then were identified by cell morphology and surface markers.After being transcripted and amplified,MUC1 mRNA was transfected into DCs by electroporation.The expression of MUC1 in DCs at different time points was detected by quantitative real-time PCR and Western blot.The survival rate of DCs before and after tramrfection was determined by MTT method.The induction of specific cytotoxic T lymphocyte (CTL) response by MUC1 mRNA transfected DCs was measured by 51Cr standard cytotoxicity test.The released amount of IFN-γ was evaluated by ELISA method.Results The cultured cells appeared typical characteristics with regard to morphology and phenotype (CD40 +,HLA-DR+,CD83 +,CD86 + ).After MUC1 mRNA transfection for 48 h,the expression of MUC1 mRNA of DCs reached the highest point ( 38.43 ) and the MUC1 protein expression also reached the highest point at 72 h.The survival rate of DCs was stabilized around 80％ after transfection.The DCs transfected with MUC1 mRNA could effectively induce HLA-A2+/MUC1 + specific CTL immune responses.Stimulated by pancreatic cancer cell line Capan-2 cells or the DCs transfected with MUC1 mRNA,the IFN-γ released in 24 h by MUC1 specific CTL were ( 28.44 ± 4.96 ) U/m1 and ( 16.31 ± 2.54) U/ml,respectively.The difference between the two groups was statistically significant (P ＜0.05 ).Conclusions DCs transfected with human pancreatic cancer MUC1 mRNA could induce CTLs and produce specific anti-tumor immunity.
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Objective To compare the effect of autologous bone marrow stem cells transplantation on liver function between first and second transplantation in decompensated liver cirrhosis patients.MethodsA total of 45 decompensated liver cirrhosis patients were enrolled, and 23patients in first transplantation group were transplanted with autologous bone marrow stem cells through femoral artery when condition was stable after medical treatment.In second transplantation group, 22 patients were accepted second transplantation in 4-12 month after the first transplantation.All the patients undergone routine blood test, congulation test and liver function examination at the fourth week and eighth week after transplantation.ResultsEight weeks after transplantation, the liver function was improved obviously in both first and second autologous bone marrow stem cells transplantation.The level of albumin in patients of second transplantation group increased from (37.26± 5.90) g/L before transplantation to (42.49 ± 4.80) g/L (P＜0.01), alanine aminotransferase (ALT) decreased from (57.05±45.51) U/L to (44.86±29.19) U/L (P＜0.05),aspartate aminotransferase (AST) decreased from (39.14-±-15.07) U/L to (53.73 ± 24.98) U/L(P＞0.05).Congulation parameters were also improved, prothrombin time (PT) decreased from (16.15±3.01) s to (14.63±2.32) s (P＜0.01), fibrinofen (Fib) increased from (2.44±0.61) g/L to (3.00±0.81) g/L (P＜0.01).Compared with first transplantation group, the albumin level was higher in second autologous bone marrow stem cells transplantation group, which increased from (38.00±6.33) g/L to (42.49±4.80) g/L (P＜0.05), AST and ALT also improved obviously, and there was significant difference between two groups.Meanwhile, Child-Pugh scores decreased from (7.22±0.67) to (6.67±[0.71) (P＜0.05).But there was no significant difference in bilirubin, FIB and PT.ConclusionThe second transplantation of autologous bone marrow stem cells could further improve liver function and maintain symptoms remission of liver cirrhosis.
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Objective To analyze the effect of autologous bone marrow stem cells transplantation in treatment of patients with decompensated cirrhosis. Methodls Seventy-eight patients (aged from 26 to 67) with decompensated cirrhosis, including 56 with hepatitis B, 21 with alcoholic cirrhosis and 1 with schistosomial cirrhosis, were included. Bone marrow was aspirated from poster superior spine. After isolation and purification, the stem cells were transplanted into liver via hepatic artery. The liver function, laboratory parameters and Child-Tureotte-Pugh scores were evaluated in 2,4 and 8 weeks after transplantation. Results At the 4th week after transplantation, the level of albumin was increased obviously from (32.9±5.58) g/L to (38.32±6.45) g/L,whereas the alanine aminotransferase was decreased from (96.92±83.91) U/L to (73.48±18.46)U/L. It was revealed that the prothrombin time was decreased from (16.66±3.91) s to (15.52±3.35) s and fibronegen increased from (2. 22 ± 0. 88) g/L to (2. 58±0. 88) g/L. After transplantation, appetite was improved in 72 cases (92.3%), ascites was decreased in 70 cases (89.7%) and abdomen distention was ameliorated in 68 cases (87.2%). There was no complications related to the transplantation. Conclusion Transplantation of autologous bone marrow stem cells is a safe and effective method in treatment of patients with decompensated cirrhosis.
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Objective To investigate the currents impact on delayed rectifier potassium (HERG)regulated by cyclooxygenase (COX)-2 in gastric cancer cells and its mechnism. Methods ① The HERG mRNA, protein and current in gastric cancer cells transfected with or without COX-2 antisense vector were measured by RT-PCR, Western blot and patch-clamp, respectively. ② cAMP concentration in gastric cancer cells transfected with or without COX-2 antisense vector was measured by ELISA. ③ The mutant HERG, which was absence of cAMP-binding domain, was constructed by PCR and transfected into gastric cancer cells. ④ The impact of COX-2 inhibitor and proglandin (PG) E2 on HERG current in gastric cancer cells transfected with or without mutant HERG was investigated by patch clamp. ⑤ The effects of agonist and antagonist of cAMP and inhibitor of protein kinase (PK) A on HERG current in gastric cancer cells transfected with or without HERG mutant were observed by patch clamp. Results ① The expression of HERG mRNA and protein in gastric cancer cells transfected with COX-2 antisense vector were not altered, but the amplitude of HERG current was diminished (P<0.05). ② The cAMP concentration in gastric cancer cells transfected with COX-2 antisense vector was lower than that in parental gastric cancer cells (P<0.05). ③ COX-2 inhibitor and PGE2 had influence on the HERG currents in gastric cancer cells. COX-2 inhibitor reduced and PGE2 enhanced the amplitude of HERG current in gastric cancer cells. However, neither COX-2 inhibitor nor PGE2 showed any negative or positive effects on currents of mutant HERG. ④ cAMP agonist enhanced the amplitude of HERG current and cAMP antagonist reduced the amplitude in gastric cancer cells. Neither agonist nor antagonist had effect on currents of mutant HERG. ⑤ PKA inhibitor did not influence the HERG current of parental gastric cancer cells and gastric cancer cells transfected with mutant. Conclusions COX-2 regulates HERG current through its catalytic product of PGE2, which binds with its receptor on the gastric cancer cells and alters cAMP level in gastric cancer cells, cAMP interacts with HERG protein by binding with cAMP-binding domain of HERG protein and exerts impact on HERG current. PKA does not participate in this process.
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Objective To observe the inhibitory effect on metastasis and growth of pancreatic cancer in mice by injection of KAI1 gene in vivo. Methods Pancreatic cancer cell line MiaPaCa Ⅱ was used to construct the nude mice models bearing tumors, then the mice were divided into normal saline group, Ad group and Ad-KAI1 group. Since the 10th days of model construction, the Ad-KAI1 was injected every 7 d and repeated twice, then the tumor size, the weight of liver, lung and their pathologic changes were evaluated. Results The tumor sizes were not significantly different between the three groups. The weight of lung and liver of Ad-KAI1 group was (0.366±0.041) g and (1. 35±0.21) g, respectively; the weight of lung and liver of Ad group was (0.57±0.065) g and (1.58±1.828) g, respectively; the weight of lung and liver of control group was (0.66±0.13)g and (1.95±0.344)g, respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 5.984, P < 0. 05), and there was no significant difference between Ad group and control group (t=1.089, P > 0.05). The number of pulmonary, liver and lymph node metastasis in Ad-KAI1 group was (1±1), (2±1) and (2±2), respectively; in Ad group was (6±2), (5 ±1), (10±2), respectively; in control group was (7±2), (6±2), (11±3), respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 7.44, 4.34, 8. 16, P < 0.05), while the difference between Ad group and control group was not significantly different (t=0.92, 0.64, 0.42, P >0.05). Conclusions KAI1 gene directly injected into tumors of nude mice may inhibit the growth and metastasis of pancreatic cancer.
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Objective To explore the value of combined biliary and duodenal stenting in the palliative treatment of pancreatic head carcinoma. Methods From August 2003 to July 2005, four patients who were diagnosed as pancreatic head carcinoma presenting obstructive jaundice were treated with biliary stenting. When duodenal obstruction occurred due to the growth of the carcinoma , duodenal stents were placed to relieve the obstruction. Results The biliary and duodenal stents were successfully positioned in all 4 patients. After biliary stents were inserted, the level of total bilirubin value of each patient decreased by 50% in 24 hours and returned to normal range within 1 week. When the biliary stents were occluded, stent replacement or restenting was performed, and relief of obstructive jaundice was thus achieved. After duodenal stents were inserted, the symptoms of duodenal obstruction were relieved and the patients were able to tolerate peroral solid food. When the duodenal obstruction recurred, a second coaxial duodenal stent was inserted. Three duodenal stents were placed in one patient because of rgrowth in size of the carcinoma. No complications related to stenting were observed. Two patients were followed up until their death with survival time of 24 months and 22 months, respectively, who were free of jaundice and duodenal obstruction at the end of their life. Two patients are still in follow-up period (16 months and 9months after stenting, respectively), who are now free of jaundice and duodenal obstruction. Conclusion Biliary stenting combined with duodenal stenting can improve the life quality of patients with pancreatic head carcinoma and is an effective palliative treatment for these patients.