Objective: To detect the mRNA expression of the cancer-testis antigens (CT) SSX1 and SSX4 gene in human hepatocellular carcinomas (HCCs) and to investigate the specificity of their expression in HCCs. Methods: The mRN...Objective: To detect the mRNA expression of the cancer-testis antigens (CT) SSX1 and SSX4 gene in human hepatocellular carcinomas (HCCs) and to investigate the specificity of their expression in HCCs. Methods: The mRNA expression of SSX1 and SSX4 in HCC tissues and the corresponding nearby liver tissues in 35 cases was detected by using RT-PCR; Six positive RT-PCR products were randomly selected and sequenced. Results: In all 35 HCC tissues, SSX1 in 27 cases (81%) and SSX4 in 23 cases (73%) were detected, and their expression was negative in the liver tissues nearby HCC and the non-tumor liver tissues (12 cirrhotic tissues and 15 normal tissues). In all 6 cases selected randomly, the results of DNA sequencing were identical with the cDNA sequence of SSX1 and SSX4 genes. The SSX1, SSX4 mRNA expression was not significantly correlated with age, sex, the tumor size, the level of tumor differentiation, the serum AFP level and the infection rate of HBV and HCV respectively (P>0.05). Conclusion: The SSX1, SSX4 mRNA expression was greatly specific in HCCs, which would not only provide the ideal target molecular sites for HCC tumor vaccines, but also establish the potential value of the polyvalent tumor-antigen vaccines for HCC therapy and its theory bases.展开更多
AIMS The CAS0 and CEA are well-described human tumor-as- sociated antigens most useful clinically in gastrointestinal cancer. In this study we compared these markers in sera from patients with malignant and benign dig...AIMS The CAS0 and CEA are well-described human tumor-as- sociated antigens most useful clinically in gastrointestinal cancer. In this study we compared these markers in sera from patients with malignant and benign digestive tract diseases. METHODS Using a side-phase radioimmunoassay,CA50 and CEA serum levels were measured in 33 control subjects and 86 patients with gastric cancer(n=34),gastric ulcer(n=27)and chronic atrophic gastritis(n=25).Carcinoma of the stomach was found in the antrum(n=22),in the body(n=3),and the fundus(n=9),and histopathologically,was divided into adeno- carcinoma(n=21),squamous cancer(n=4)and not divided (n=9).Gastric ulcer,when present,appeared in the antrun(18 patients),the body(3)and the fundus(9)and chronic atrophic gastritis was all associated with intestinal metaplasia(IM). RESULTS The normal ranges established for CA50 and CEA in the control group were 16.26+6.14 kU/L and 3.12±1.03/μg/L respectively.In the patients with gastric cancer,serum levels of CA50(112.67±38.36 kU/L)and CEA(10.28±3.76μg/L) were elevated significantly(P<0.01,respectively),the former being>22 kU/L in 18 of 34 patients(53%;range,5-1 550 kU/ L),and the latter>5 μg/L in 19 of 34 patients(55.8%,range, 0.5-17.4 μg/L).A statistically significant correlation was found between the levels of CA50 and GEA(r=0.648,P<0.01). The serum levels of CA50(46.4±25.9 kU/L,P<0.01 )and CEA(6.85±2.43 μg/L,P<0.01)were much lower in patients with gastric ulcer or chronic atrophic gastritis(P>0.05). CONCLUSIONS Based on these results,it is concluded that CA50 and CEA are indicators for advanced gastric cancer,and af- ter surgery,their serum levels may decrease considerably. Overall,there is such a close correlation between them that in clinical practice they might be of great value to the diagnosis of gastric cancer.展开更多
Aim To reveal the main active components and the action mechanisms of Radix astragali on insulin sensitivity improvement, we have investigated the effects of polysaccharide portion and saponin portion of Radix astraga...Aim To reveal the main active components and the action mechanisms of Radix astragali on insulin sensitivity improvement, we have investigated the effects of polysaccharide portion and saponin portion of Radix astragali extracts on blood biochemical indices and related gene expression of dexamethasone-induced SD rats. Methods SD rats (6 per group) received 2 μg/day subcutaneous dexamethasone for 4 weeks plus same dose (10 g material/kg) of polysaccharide or saponin extracts of Radix astragali. Blood samples, kidney tissues and epididymal fat pads were taken at the end of the experiment. Serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDLC), high density lipoprotein cholesterol (HDLC), glucose (GLU) and insulin (INS) levels were measured, respectively, mRNA levels of angiotensinogen in kidney, adiponectin and leptin as well as TNF-α in epididymal fats were determined by RT-PCR assay using GAPDH gene as an internal control. Results Both of polysaccharide and saponin extracts of Radix astragali exhibited positive effects in reducing serum triglycerides, glucose, and insulin levels of dexamethasone-induced SD rats. The saponin group showed more improvements on quantitive insulin sensitivity check index (QUICKI) than the polysaccharide group did. Both of the extracts down-regulated kidney angiotensinogen and fat TNF-α mRNA levels while they were simultaneously up-regulating fat adiponectin and leptin mRNA levels. No significant difference was found between actions of the two extracts. Conclusion Both of polysaccharide and saponin extracts of Radix astragali can improve insulin sensitivity. This action might be closely related to down-regulation of angiotensinogen, TNF-α and up-regulation of adiponectin and leptin expression. The results partly explained the improvement of type Ⅱ diabetes and diabetic nephropathy by Radix astragali. The similar actions of the two crude extracts suggest that unknown key active compounds might exist in both and remain to be discovered.展开更多
AIM: To evaluate the differences that exist bet- ween peripheral and mesenteric serum levels of carcinoembryonic antigen (CEA) and cytokeratins in patients with colorectal adenocarcinoma. METHODS: One hundred and ...AIM: To evaluate the differences that exist bet- ween peripheral and mesenteric serum levels of carcinoembryonic antigen (CEA) and cytokeratins in patients with colorectal adenocarcinoma. METHODS: One hundred and thirty-eight patients with colorectal adenocarcinoma who underwent surgery at Hospital Sao Paulo (Discipline of Surgical Gastroenterology of UNIFESP-EPM) between December 1993 and March 2000 were retrospectively analyzed. Differences between CEA and cytokeratin (TPA-M) levels in peripheral blood (P) and in mesenteric blood (M) were studied. Associations were investigated between peripheral and mesenteric levels and the staging and histopathological variables (degree of cell differentiation, macroscopic appearance, tumor dimensions and presence of lymphatic and venous invasion). RESULTS: Differences were observed in the numerical values of the marker levels: CEA (M) (39.10 mg/1 ± 121.19 mg/L) vs CEA (P) (38.5 mg/L ± 122.55 mg/L), P 〈 0.05; TPA-M (M) (325.06 U/L ±527.29 U/L) vs TPA-M (P) (279.48 U/L ±455.81 U/L), P 〈 0.01. The mesenteric CEA levels were higher in more advanced tumors (P 〈 0.01), in vegetating lesions (34.44 mg/L ± 93.07 mg/L) (P 〈 0.01) and with venous invasion (48.41 mg/L ± 129.86 mg/L) (P 〈 0.05). Peripheral CEA was higher with more advanced staging (P 〈 0.01)and in lesions with venous invasion (53.23 mg/L ± 258.57 mg/L) (P 〈 0.05). The patients demonstrated increased mesenteric and peripheral TPA-M levels with more advanced tumors (P 〈 0.01 and P 〈 0.01) and in non-ulcerated lesions [530.45 U/L =1= 997.46 U/L (P 〈 0.05) and 457.95 U/L ± 811.36 U/L (P 〈 0.01)]. CONCLUSION: The mesenteric levels of the tumor markers CEA and cytokeratins were higher than the peripheral levels in these colorectal adenocarcinoma patients, Higher levels of these biologic tumor markers are associated with an advanced state of cancerous dissemination展开更多
Nasopharyngeal carcinoma (NPC) is a common cancer in Southern China and Southeast Asia. The disease is a poorly differentiated carcinoma without effective cure, and the mechanism underlying its development remains l...Nasopharyngeal carcinoma (NPC) is a common cancer in Southern China and Southeast Asia. The disease is a poorly differentiated carcinoma without effective cure, and the mechanism underlying its development remains largely unknown. Of several factors identified in NPC aetiology in recent years, Epstein-Barr virus (EBV) infection has emerged to be most important. In almost all NPC cells, EBV uses several intracellular mechanisms to cause oncogenic evolution of the infected cells. One such mechanism by which EBV infection induces cellular immortalization is believed to be through the activation of telomerase, an enzyme that is normally repressed but becomes activated during cancer development. Studies show that greater than 85% of primary NPC display high telomerase activity by mechanisms involving EBV infection, consistent with the notion that EBV is commonly involved in inducing cell immortalization. More recently, different EBV proteins have been shown to activate or inhibit the human telomerase reverse transcriptase gene, by modulating intracellular signalling pathways. These findings suggest a new model with a number of challenges towards our understanding, molecular targeting and therapeutic intervention in NPC.展开更多
The c-erbB-2 proto-oncogene encodes a 185kDa protein p!85, which belongs to epidermal growth factor receptor family. Amplification of this gene has been shown to correlate with poor clinical prognosis for certain canc...The c-erbB-2 proto-oncogene encodes a 185kDa protein p!85, which belongs to epidermal growth factor receptor family. Amplification of this gene has been shown to correlate with poor clinical prognosis for certain cancer patients. The monoclonal antibody A21 which directed against p185 specifically inhibits proliferation of tumor cells overexpressing p185, hence allows it to be a candidate for targeted therapy. In order to overcome several drawbacks of murine MAb, we cloned its VH and VL genes and constructed the single-chain Fv (scFv) through a peptide linker. The recombinant scFvA21 was expressed in Escherichia coli and purified by the affinity column. Subsequently it was characterized by ELISA, Western blot, cell immunohistochemistry and FACS. All these assays showed the binding activity to extracellular domain (ECD) of p!85. Based on those properties of scFvA21, we further constructed the scFv-Fc fusion molecule with a homodimer form and the recombinant product was expressed in mammalian cells. In a series of subsequent analysis this fusion protein showed identical antigen binding site and activity with the parent antibody. These anti-p185 engineered antibodies have promised to be further modified as a tumor targeting drugs, with a view of application in the diagnosis and treatment of human breast cancer.展开更多
AIM: To investigate the expression of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) in metastatic lymph nodes from gastrointestinal cancer. METHODS: Metastatic lymph nodes from gastrointestina ca...AIM: To investigate the expression of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) in metastatic lymph nodes from gastrointestinal cancer. METHODS: Metastatic lymph nodes from gastrointestina cancer were detected for RCAS1 by immunohistochemica staining and mRNA in situ hybridization.RESULTS: A total of 102 metastatic lymph nodes from bile duct, gastric, colon, and pancreatic cancer were investigated for RCAS1 expression. The immunoreactivity of RCAS1 was identified in 100% of metastatic lymph nodes. Both local and distant metastatic lymph nodes showed RCAS1 expression. On the contrary, specimens of non-cancerous lymph nodes were negative for RCAS1. The result of mRNA in situ hybridization was also confirmed by the finding of immunohistochemical staining. RCAS1 mRNA was detected in all tumor cells that metastasized to lymph nodes. CONCLUSION: All metastatic lymph nodes express RCAS1 in tumor cells at both protein and mRNA levels, and RCAS1 should be used as a complementary factor for identification of metastatic lymph nodes from gastrointestinal cancers.展开更多
AIM: To purify and characterizeα-L-fucosidase from human liver cancer tissue and to detect the localization ofα-L-fucosidase in tumor tissue. METHODS: Cation exchange chromatography on CM-52 and ultrafiltration were...AIM: To purify and characterizeα-L-fucosidase from human liver cancer tissue and to detect the localization ofα-L-fucosidase in tumor tissue. METHODS: Cation exchange chromatography on CM-52 and ultrafiltration were used to separateα-Lfucosidase (AFU) from crude extract of liver cancer tissue. 4-methylumbelliferyl-α-L-fucopyranoside was used as a fluorescent substrate to quantify the purified AFU activity in each step. A polyclonal antibody (pAb) against the purified AFU was obtained by anion exchange chromatography on DEAE-52 after ammonium sulfate fractionation and ultrafiltration. Immuohistochemical staining was used to observe the expression of AFU in malignant and adjacent liver tissues. RESULTS: Humanα-L-fucosidase was purified 74-fold to apparent homogeneity with 15% yield. SDSPAGE indicated the presence of one subunit of molecular weight of 55 Ku. The specific activity of AFU in pooled fraction by chromatography was 10085 IU/mg. Western blot analysis indicated that the pAb could recognize one protein band of molecular weight of 55 Ku. The expression of AFU was observed in cytoplasm membrane of liver cancer tissue but not in that of adjacent tissue. CONCLUSION: The purifiedα-L-fucosidase from primary hepatocarcinoma (PHC) is different in its properties fromα-L-fucosidase in human other organs. The polyclonal antibody prepared in this experiment can be applied to the diagnosis of PHC.展开更多
Anticancer immunotherapy has undergone a long evolving journey for decades, and has been dramatically applied to mainstream treatments in oncology in recent 5 years. This progress represents an advanced milestone foll...Anticancer immunotherapy has undergone a long evolving journey for decades, and has been dramatically applied to mainstream treatments in oncology in recent 5 years. This progress represents an advanced milestone following cytotoxic medicine and targeted therapy. Cellular immunity plays a pivotal role in the immune responses of hosts to tumor antigens. Such immunity is notably suppressed during neoplastic progression due to immuno-editing processes. Cellular immunity can also be selectively reactivated to combat malignancies while exploiting the advantages of contemporary scientific breakthroughs in molecular immunology and genetic engineering. The rapid advancement of cellular immunity-based therapeutic approaches has achieved high efficacy in certain cancer patients. Consequently, the landscape of oncologic medicine and pharmaceutical innovation has transformed recently. In this regard, we present a comprehensive update on clinically established anti-cancer treatments with cell immunity augmentation as the major mechanism of action.展开更多
AIM: To examine the concentration of a new antigen SC6 (SC6-Ag) recognized by monoclonal antibody (MAb)in patients with pancreatic cancer and other malignant or benign diseases and to understand whether SC6-Ag has any...AIM: To examine the concentration of a new antigen SC6 (SC6-Ag) recognized by monoclonal antibody (MAb)in patients with pancreatic cancer and other malignant or benign diseases and to understand whether SC6-Ag has any clinical significance in distinguishing pancreatic cancer from other gastrointestinal diseases.METHODS: Six hundred and ninety-five serum specimens obtained from 115 patients with pancreatic cancer, 154 patients with digestive cancer and 95patients with non-digestive cancer were used and classified in this study. Serum specimens obtained from 140 patients with benign digestive disease and 89 patients with non-benign digestive disease served as controls. Ascites was tapped from 16 pancreatic cancer patients, 19 hepatic cancer patients, 16 colonic cancer patients, 10 gastric cancer and 6 severe necrotic pancreatitis patients. The samples were quantitated by solid-phase radioimmunoassay. The cut-off values (CV)of 41, 80, and 118 U/mL were used.RESULTS: The average intra- and interassay CV detected by immunoradiometric assay of SC6-Ag was 5.4% and 8.7%, respectively. The sensitivity and specificity were 73.0% and 90.9% respectively. The levels in most malignant and benign cases were within the normal upper limit. Among the 16 pancreatic cancer cases, the concentration of SC6-Ag in ascites was over the normal range in 93.8% patients. There was no significant difference in the concentration of SC6-Ag.Decreased expression of SC6-Ag in sera was significantly related to tumor differentiation. The concentration of SC6-Ag was higher in patients before surgery than after surgery. The specificity of SC6-Ag and CA19-9 was significantly higher than that of ultrasound and computer tomography (CT) in pancreatic cancer patients. Higher positive predictive values were indicated in 92.3% SC6-Ag and 88.5% CA19-9, but lower in 73.8% ultrasound and 76.2% CT.CONCLUSION: The combined test of SC6-Ag and CA19-9 may improve the diagnostic rate of primary cancer. The detection of SC6-Ag is valuable in the diagnosis of pancreatic cancer before and after surgery.展开更多
Extrathoracic solitary fibrous tumors(SFTs) have been described at almost every anatomic location of human body,but reports of SFT in the abdominal cavity are rare.We herein present a rare case of SFT originating from...Extrathoracic solitary fibrous tumors(SFTs) have been described at almost every anatomic location of human body,but reports of SFT in the abdominal cavity are rare.We herein present a rare case of SFT originating from greater omentum.Computed tomography revealed a 15.8 cm × 21.0 cm solid mass located at superior aspect of stomach.Open laparotomy confirmed its mesenchymal origin.Microscopically,its tissue was composed of non-organized and spindle-shaped cells exhibiting atypical nuclei,which were divided up by branching vessel and collagen bundles.Immunohistochemical staining showed that this tumor was negative for CD117,CD99,CD68,cytokeratin,calretinin,desmin,epithelial membrane antigen,F8 and S-100,but positive for CD34,bcl-2,α-smooth muscle actin and vimentin.The patient presented no evidence of recurrence during follow-up.SFT arising from abdominal cavity can be diagnosed by histological findings and immunohistochemical markers,especially for CD34 and bcl-2 positive cases.展开更多
AIMS To evaluate the sensitivity,specificity and clinical value of hepatic carcinoma associated mem- brane protein antigen (HAg18-1) used as a reagent of serum quick enzyme linked immunosorbent assay (ELISA) for the d...AIMS To evaluate the sensitivity,specificity and clinical value of hepatic carcinoma associated mem- brane protein antigen (HAg18-1) used as a reagent of serum quick enzyme linked immunosorbent assay (ELISA) for the detection of primary hepatocellular car- cinoma (PHCC). METHODS Serum quick enzyme linked immunosor- bent assay (ELISA) with HAg18-1 as reagent,which was prepared by monoclonal antibody against human hepatic carcinoma,was performed on 100 cases of pri- mary hepatocellular carcinoma (PHCC),5 cases of hepatic biliary carcinoma (HBC),10 cases of metastatic hepatic carcinoma (MHC),20 cases of hep- atitis B (HB),20 cases of liver cirrhosis (LC),20 cas- es of gastrointestinal malignant tumours and 20 cases of gastronintestinal inflammatory diseases (including ulcers). Alpha-fetoprotein (AFP) detection concurrent- ly for each case. Twenty samples of bank blood were tested as controls. RESULTS Positive rate of HAg18-1 ELISA and AFP detection were 81% and 68% in PHCC,20% and 40% in HBC,19% and 20% in MHC,10% and 20% in BH, 10% and 20% in LC,10% and 15% in gastrointestinal malignant tumors,and 5% and 10% in gastrointestinal inflammatory diseases. No positive result of HAg18-1 ELISA and AFP detection occurred in any sample of bank blood. CONCLUSIONS HAg18-1 ELISA has high sensitivity and specificity in the detection of PHCC. HAg18-1 ELISA and AFP detection,if used together,may be complementary to each other in the diagnosis of PHCC.展开更多
AIM: To investigate the usefulness of tumor markers and adenosine deaminase in differentiating between tuberculous peritonitis (TBP) and peritoneal carcinoma- tosis (PC). METHODS: A retrospective analysis of dat...AIM: To investigate the usefulness of tumor markers and adenosine deaminase in differentiating between tuberculous peritonitis (TBP) and peritoneal carcinoma- tosis (PC). METHODS: A retrospective analysis of data was per- formed on consecutive patients who underwent perito- neoscopic and abdominal computed tomography (CT) evaluations. Among 75 patients at the Seoul National University Hospital from January 2000 to June 2010 who underwent both tests, 27 patients (36.0%) and 25 patients (33.3%) were diagnosed with TBP and PC, re- spectively. Diagnosis was confirmed by peritoneoscopic biopsy. RESULTS: Serum c-reactive protein (7.88:1:6.62 mg/ dL vs 3.12 + 2.69 mg/dL, P = 0.01), ascites adenos- ine deaminase (66.76:1:32.09 IU/L vs 13.89 :l: 8.95 IU/L, P 〈 0.01), ascites lymphocyte proportion (67.77 :1: 23.41% vs 48.36 + 18.78%, P 〈 0.01), and serum- ascites albumin gradient (0.72 + 0.49 g/dL vs 1.05 + 0.50 g/dL, P = 0.03) were significantly different be- tween the two groups. Among tumor markers, serum and ascites carcinoembryonic antigen, serum carbohy- drate antigen 19-9 showed significant difference be- tween two groups. Abdominal CT examinations showed that smooth involvement of the parietal peritoneum was more common in the TBP group (77.8% vs 40.7%) whereas nodular involvement was more common in the PC group (14.8% vs 40.7%, P = 0.04). From receiver operating characteristic (ROC) curves ascites adeno- sines deaminase (ADA) showed better discriminative capability than tumor markers. An ADA cut-off level of 21 IU/L was found to yield the best results of differ- ential diagnosis; sensitivity, specificity, positive predic- tive value, and negative predictive value were 92.0%, 85.0%, 88.5% and 89.5%, respectively. CONCLUSION: Besides clinical and radiologic findings, ascitic fluid ADA measurement is helpful in the differen- tial diagnosis of TBP and PC.展开更多
AIM:To compare the outcome of surgical treatment of colorectal adenocarcinoma in elderly and younger patients.METHODS:The outcomes of 122 patients with colorectal adenocarcinoma who underwent surgical treatment betwee...AIM:To compare the outcome of surgical treatment of colorectal adenocarcinoma in elderly and younger patients.METHODS:The outcomes of 122 patients with colorectal adenocarcinoma who underwent surgical treatment between January 2004 and June 2009 were analyzed.The clinicopathological and blood biochemistry data of the younger group(<75 years) and the elderly group (≥75 years) were compared.RESULTS:There were no significant differences between the two groups in operation time,intraoperative blood loss,hospital stay,time to resumption of oral intake,or morbidity.The elderly group had a significantly higher rate of hypertension and cardiovascular disease.The perioperative serum total protein and albumin levels were significantly lower in the elderly than in the younger group.The serum carcinoembryonic antigen level was lower in the elderly than in the younger group,and there was a significant decreasing trend after the operation in the elderly group.CONCLUSION:The short-term outcomes of surgical treatment in elderly patients with colorectal adenocarcinoma were acceptable.Surgical treatment in elderly patients was considered a selectively effective approach.展开更多
The data from in vitro and animal experiment study has showed that costimulaory molecule B7 1 plays an important role in antitumor immunity In the present study, B7 1 expression was ob...The data from in vitro and animal experiment study has showed that costimulaory molecule B7 1 plays an important role in antitumor immunity In the present study, B7 1 expression was observed in 130 samples from a veriety of human malignancies by using immunocytochemistry, in situ hybridization and RT PCR combined with dot hybridization and B7 1 specific Mab and probe The results demonstrated B7 1 expression on tumor cells in 76 cases at both protein and mRNA level Forty two specimens were stained with B7 1 HLA ABC and HLA DR Mab and 26 showed that the three antibodies used all were positive Together with the achievement in tumor antigen study, the present findings imply that in most tumors (if not all) the tumor cells have all the requisite element to elicit anti tumor rejection response, the heterogeneous mechanism for tumor escape from immunosurvillance should be emphasized展开更多
AIM:To evaluate pretreatment serum carcinoembryonic antigen(CEA) as a predictor of survival for patients with locally advanced gastric cancer receiving perioperative chemotherapy.METHODS:We retrospectively studied a c...AIM:To evaluate pretreatment serum carcinoembryonic antigen(CEA) as a predictor of survival for patients with locally advanced gastric cancer receiving perioperative chemotherapy.METHODS:We retrospectively studied a cohort of 228 gastric cancer patients who underwent D2 gastrectomy combined with chemotherapy at the Sun Yat-sen University Cancer Center between January 2005 and December 2009.Among them,168 patients received 6-12 cycles of oxaliplatin-based adjuvant(post-operative) chemotherapy,while 60 received perioperative chemotherapy(2 cycles of FOLFOX6 or XELOX before surgery and 4-10 cycles after surgery).Serum CEA was measured using an enzyme immunoassay.The followup lasted until December 2010.RESULTS:In the group that had elevated serum CEA,the difference in survival time between patients receiving perioperative chemotherapy and those receiving adjuvant chemotherapy had no statistical significance(P > 0.05).However,in the group that had normal serum CEA,patients receiving perioperative chemotherapy had a longer survival time.In multivariate analysis,T staging and lymph node metastatic rate were independent prognostic factors for the patients.Perioperative chemotherapy improved the overall survival of patients who had a normal pretreatment CEA level(P = 0.070).CONCLUSION:Normal pretreatment serum CEA is a predictor of survival for patients receiving perioperative chemotherapy.展开更多
We have developed and tested chimeric T-cell receptors (TCR) specific for p185HER2. In these experiments, retroviral vectors expressing the N29γ or N29ζ receptors were constructed in pRET6. Amphotropic viral produce...We have developed and tested chimeric T-cell receptors (TCR) specific for p185HER2. In these experiments, retroviral vectors expressing the N29γ or N29ζ receptors were constructed in pRET6. Amphotropic viral producer cells were established in the GALV-based PG13 packaging cell line. Ficoll purified human peripheral blood lymphocytes (PBL) were virally transduced using an optimized protocol incorporating activation with immobilized anti-CD3/anti-CD28 monoclonal anti- bodies, followed by viral infection in the presence of fibronectin fragment CH296. Transduced cells were co-cultured with human tumor cell lines that overexpress (SK-OV-3) or underexpress (MCF7) p185HER2 to assay for antigen specific im- mune responses. Both CM+ and CD8+ T-cells transduced with the N29γ or N29ζ chTCR demonstrated HER2-specific anti- gen responses, as determined by release of Th1 like cytokines, and cellular cytotoxicity assays. Our results support the fea- sibility of adoptive immunotherapy with genetically modified T-cells expressing a chTCR specific for p185HER2.展开更多
文摘Objective: To detect the mRNA expression of the cancer-testis antigens (CT) SSX1 and SSX4 gene in human hepatocellular carcinomas (HCCs) and to investigate the specificity of their expression in HCCs. Methods: The mRNA expression of SSX1 and SSX4 in HCC tissues and the corresponding nearby liver tissues in 35 cases was detected by using RT-PCR; Six positive RT-PCR products were randomly selected and sequenced. Results: In all 35 HCC tissues, SSX1 in 27 cases (81%) and SSX4 in 23 cases (73%) were detected, and their expression was negative in the liver tissues nearby HCC and the non-tumor liver tissues (12 cirrhotic tissues and 15 normal tissues). In all 6 cases selected randomly, the results of DNA sequencing were identical with the cDNA sequence of SSX1 and SSX4 genes. The SSX1, SSX4 mRNA expression was not significantly correlated with age, sex, the tumor size, the level of tumor differentiation, the serum AFP level and the infection rate of HBV and HCV respectively (P>0.05). Conclusion: The SSX1, SSX4 mRNA expression was greatly specific in HCCs, which would not only provide the ideal target molecular sites for HCC tumor vaccines, but also establish the potential value of the polyvalent tumor-antigen vaccines for HCC therapy and its theory bases.
文摘AIMS The CAS0 and CEA are well-described human tumor-as- sociated antigens most useful clinically in gastrointestinal cancer. In this study we compared these markers in sera from patients with malignant and benign digestive tract diseases. METHODS Using a side-phase radioimmunoassay,CA50 and CEA serum levels were measured in 33 control subjects and 86 patients with gastric cancer(n=34),gastric ulcer(n=27)and chronic atrophic gastritis(n=25).Carcinoma of the stomach was found in the antrum(n=22),in the body(n=3),and the fundus(n=9),and histopathologically,was divided into adeno- carcinoma(n=21),squamous cancer(n=4)and not divided (n=9).Gastric ulcer,when present,appeared in the antrun(18 patients),the body(3)and the fundus(9)and chronic atrophic gastritis was all associated with intestinal metaplasia(IM). RESULTS The normal ranges established for CA50 and CEA in the control group were 16.26+6.14 kU/L and 3.12±1.03/μg/L respectively.In the patients with gastric cancer,serum levels of CA50(112.67±38.36 kU/L)and CEA(10.28±3.76μg/L) were elevated significantly(P<0.01,respectively),the former being>22 kU/L in 18 of 34 patients(53%;range,5-1 550 kU/ L),and the latter>5 μg/L in 19 of 34 patients(55.8%,range, 0.5-17.4 μg/L).A statistically significant correlation was found between the levels of CA50 and GEA(r=0.648,P<0.01). The serum levels of CA50(46.4±25.9 kU/L,P<0.01 )and CEA(6.85±2.43 μg/L,P<0.01)were much lower in patients with gastric ulcer or chronic atrophic gastritis(P>0.05). CONCLUSIONS Based on these results,it is concluded that CA50 and CEA are indicators for advanced gastric cancer,and af- ter surgery,their serum levels may decrease considerably. Overall,there is such a close correlation between them that in clinical practice they might be of great value to the diagnosis of gastric cancer.
文摘Aim To reveal the main active components and the action mechanisms of Radix astragali on insulin sensitivity improvement, we have investigated the effects of polysaccharide portion and saponin portion of Radix astragali extracts on blood biochemical indices and related gene expression of dexamethasone-induced SD rats. Methods SD rats (6 per group) received 2 μg/day subcutaneous dexamethasone for 4 weeks plus same dose (10 g material/kg) of polysaccharide or saponin extracts of Radix astragali. Blood samples, kidney tissues and epididymal fat pads were taken at the end of the experiment. Serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDLC), high density lipoprotein cholesterol (HDLC), glucose (GLU) and insulin (INS) levels were measured, respectively, mRNA levels of angiotensinogen in kidney, adiponectin and leptin as well as TNF-α in epididymal fats were determined by RT-PCR assay using GAPDH gene as an internal control. Results Both of polysaccharide and saponin extracts of Radix astragali exhibited positive effects in reducing serum triglycerides, glucose, and insulin levels of dexamethasone-induced SD rats. The saponin group showed more improvements on quantitive insulin sensitivity check index (QUICKI) than the polysaccharide group did. Both of the extracts down-regulated kidney angiotensinogen and fat TNF-α mRNA levels while they were simultaneously up-regulating fat adiponectin and leptin mRNA levels. No significant difference was found between actions of the two extracts. Conclusion Both of polysaccharide and saponin extracts of Radix astragali can improve insulin sensitivity. This action might be closely related to down-regulation of angiotensinogen, TNF-α and up-regulation of adiponectin and leptin expression. The results partly explained the improvement of type Ⅱ diabetes and diabetic nephropathy by Radix astragali. The similar actions of the two crude extracts suggest that unknown key active compounds might exist in both and remain to be discovered.
文摘AIM: To evaluate the differences that exist bet- ween peripheral and mesenteric serum levels of carcinoembryonic antigen (CEA) and cytokeratins in patients with colorectal adenocarcinoma. METHODS: One hundred and thirty-eight patients with colorectal adenocarcinoma who underwent surgery at Hospital Sao Paulo (Discipline of Surgical Gastroenterology of UNIFESP-EPM) between December 1993 and March 2000 were retrospectively analyzed. Differences between CEA and cytokeratin (TPA-M) levels in peripheral blood (P) and in mesenteric blood (M) were studied. Associations were investigated between peripheral and mesenteric levels and the staging and histopathological variables (degree of cell differentiation, macroscopic appearance, tumor dimensions and presence of lymphatic and venous invasion). RESULTS: Differences were observed in the numerical values of the marker levels: CEA (M) (39.10 mg/1 ± 121.19 mg/L) vs CEA (P) (38.5 mg/L ± 122.55 mg/L), P 〈 0.05; TPA-M (M) (325.06 U/L ±527.29 U/L) vs TPA-M (P) (279.48 U/L ±455.81 U/L), P 〈 0.01. The mesenteric CEA levels were higher in more advanced tumors (P 〈 0.01), in vegetating lesions (34.44 mg/L ± 93.07 mg/L) (P 〈 0.01) and with venous invasion (48.41 mg/L ± 129.86 mg/L) (P 〈 0.05). Peripheral CEA was higher with more advanced staging (P 〈 0.01)and in lesions with venous invasion (53.23 mg/L ± 258.57 mg/L) (P 〈 0.05). The patients demonstrated increased mesenteric and peripheral TPA-M levels with more advanced tumors (P 〈 0.01 and P 〈 0.01) and in non-ulcerated lesions [530.45 U/L =1= 997.46 U/L (P 〈 0.05) and 457.95 U/L ± 811.36 U/L (P 〈 0.01)]. CONCLUSION: The mesenteric levels of the tumor markers CEA and cytokeratins were higher than the peripheral levels in these colorectal adenocarcinoma patients, Higher levels of these biologic tumor markers are associated with an advanced state of cancerous dissemination
文摘Nasopharyngeal carcinoma (NPC) is a common cancer in Southern China and Southeast Asia. The disease is a poorly differentiated carcinoma without effective cure, and the mechanism underlying its development remains largely unknown. Of several factors identified in NPC aetiology in recent years, Epstein-Barr virus (EBV) infection has emerged to be most important. In almost all NPC cells, EBV uses several intracellular mechanisms to cause oncogenic evolution of the infected cells. One such mechanism by which EBV infection induces cellular immortalization is believed to be through the activation of telomerase, an enzyme that is normally repressed but becomes activated during cancer development. Studies show that greater than 85% of primary NPC display high telomerase activity by mechanisms involving EBV infection, consistent with the notion that EBV is commonly involved in inducing cell immortalization. More recently, different EBV proteins have been shown to activate or inhibit the human telomerase reverse transcriptase gene, by modulating intracellular signalling pathways. These findings suggest a new model with a number of challenges towards our understanding, molecular targeting and therapeutic intervention in NPC.
基金This work was supported by funds of Natural Science of Scientific Committee and Educational Committee of AN-HUI Province respectively, and Hi-tech Research and Development Program ("863" Program).
文摘The c-erbB-2 proto-oncogene encodes a 185kDa protein p!85, which belongs to epidermal growth factor receptor family. Amplification of this gene has been shown to correlate with poor clinical prognosis for certain cancer patients. The monoclonal antibody A21 which directed against p185 specifically inhibits proliferation of tumor cells overexpressing p185, hence allows it to be a candidate for targeted therapy. In order to overcome several drawbacks of murine MAb, we cloned its VH and VL genes and constructed the single-chain Fv (scFv) through a peptide linker. The recombinant scFvA21 was expressed in Escherichia coli and purified by the affinity column. Subsequently it was characterized by ELISA, Western blot, cell immunohistochemistry and FACS. All these assays showed the binding activity to extracellular domain (ECD) of p!85. Based on those properties of scFvA21, we further constructed the scFv-Fc fusion molecule with a homodimer form and the recombinant product was expressed in mammalian cells. In a series of subsequent analysis this fusion protein showed identical antigen binding site and activity with the parent antibody. These anti-p185 engineered antibodies have promised to be further modified as a tumor targeting drugs, with a view of application in the diagnosis and treatment of human breast cancer.
基金Supported by the Thailand Research Fund in the Royal Golden Jubilee Program
文摘AIM: To investigate the expression of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) in metastatic lymph nodes from gastrointestinal cancer. METHODS: Metastatic lymph nodes from gastrointestina cancer were detected for RCAS1 by immunohistochemica staining and mRNA in situ hybridization.RESULTS: A total of 102 metastatic lymph nodes from bile duct, gastric, colon, and pancreatic cancer were investigated for RCAS1 expression. The immunoreactivity of RCAS1 was identified in 100% of metastatic lymph nodes. Both local and distant metastatic lymph nodes showed RCAS1 expression. On the contrary, specimens of non-cancerous lymph nodes were negative for RCAS1. The result of mRNA in situ hybridization was also confirmed by the finding of immunohistochemical staining. RCAS1 mRNA was detected in all tumor cells that metastasized to lymph nodes. CONCLUSION: All metastatic lymph nodes express RCAS1 in tumor cells at both protein and mRNA levels, and RCAS1 should be used as a complementary factor for identification of metastatic lymph nodes from gastrointestinal cancers.
基金Supported by the National High Technology Research and Development Program of China (863 Program), No.2002AA2Z2011
文摘AIM: To purify and characterizeα-L-fucosidase from human liver cancer tissue and to detect the localization ofα-L-fucosidase in tumor tissue. METHODS: Cation exchange chromatography on CM-52 and ultrafiltration were used to separateα-Lfucosidase (AFU) from crude extract of liver cancer tissue. 4-methylumbelliferyl-α-L-fucopyranoside was used as a fluorescent substrate to quantify the purified AFU activity in each step. A polyclonal antibody (pAb) against the purified AFU was obtained by anion exchange chromatography on DEAE-52 after ammonium sulfate fractionation and ultrafiltration. Immuohistochemical staining was used to observe the expression of AFU in malignant and adjacent liver tissues. RESULTS: Humanα-L-fucosidase was purified 74-fold to apparent homogeneity with 15% yield. SDSPAGE indicated the presence of one subunit of molecular weight of 55 Ku. The specific activity of AFU in pooled fraction by chromatography was 10085 IU/mg. Western blot analysis indicated that the pAb could recognize one protein band of molecular weight of 55 Ku. The expression of AFU was observed in cytoplasm membrane of liver cancer tissue but not in that of adjacent tissue. CONCLUSION: The purifiedα-L-fucosidase from primary hepatocarcinoma (PHC) is different in its properties fromα-L-fucosidase in human other organs. The polyclonal antibody prepared in this experiment can be applied to the diagnosis of PHC.
文摘Anticancer immunotherapy has undergone a long evolving journey for decades, and has been dramatically applied to mainstream treatments in oncology in recent 5 years. This progress represents an advanced milestone following cytotoxic medicine and targeted therapy. Cellular immunity plays a pivotal role in the immune responses of hosts to tumor antigens. Such immunity is notably suppressed during neoplastic progression due to immuno-editing processes. Cellular immunity can also be selectively reactivated to combat malignancies while exploiting the advantages of contemporary scientific breakthroughs in molecular immunology and genetic engineering. The rapid advancement of cellular immunity-based therapeutic approaches has achieved high efficacy in certain cancer patients. Consequently, the landscape of oncologic medicine and pharmaceutical innovation has transformed recently. In this regard, we present a comprehensive update on clinically established anti-cancer treatments with cell immunity augmentation as the major mechanism of action.
文摘AIM: To examine the concentration of a new antigen SC6 (SC6-Ag) recognized by monoclonal antibody (MAb)in patients with pancreatic cancer and other malignant or benign diseases and to understand whether SC6-Ag has any clinical significance in distinguishing pancreatic cancer from other gastrointestinal diseases.METHODS: Six hundred and ninety-five serum specimens obtained from 115 patients with pancreatic cancer, 154 patients with digestive cancer and 95patients with non-digestive cancer were used and classified in this study. Serum specimens obtained from 140 patients with benign digestive disease and 89 patients with non-benign digestive disease served as controls. Ascites was tapped from 16 pancreatic cancer patients, 19 hepatic cancer patients, 16 colonic cancer patients, 10 gastric cancer and 6 severe necrotic pancreatitis patients. The samples were quantitated by solid-phase radioimmunoassay. The cut-off values (CV)of 41, 80, and 118 U/mL were used.RESULTS: The average intra- and interassay CV detected by immunoradiometric assay of SC6-Ag was 5.4% and 8.7%, respectively. The sensitivity and specificity were 73.0% and 90.9% respectively. The levels in most malignant and benign cases were within the normal upper limit. Among the 16 pancreatic cancer cases, the concentration of SC6-Ag in ascites was over the normal range in 93.8% patients. There was no significant difference in the concentration of SC6-Ag.Decreased expression of SC6-Ag in sera was significantly related to tumor differentiation. The concentration of SC6-Ag was higher in patients before surgery than after surgery. The specificity of SC6-Ag and CA19-9 was significantly higher than that of ultrasound and computer tomography (CT) in pancreatic cancer patients. Higher positive predictive values were indicated in 92.3% SC6-Ag and 88.5% CA19-9, but lower in 73.8% ultrasound and 76.2% CT.CONCLUSION: The combined test of SC6-Ag and CA19-9 may improve the diagnostic rate of primary cancer. The detection of SC6-Ag is valuable in the diagnosis of pancreatic cancer before and after surgery.
文摘Extrathoracic solitary fibrous tumors(SFTs) have been described at almost every anatomic location of human body,but reports of SFT in the abdominal cavity are rare.We herein present a rare case of SFT originating from greater omentum.Computed tomography revealed a 15.8 cm × 21.0 cm solid mass located at superior aspect of stomach.Open laparotomy confirmed its mesenchymal origin.Microscopically,its tissue was composed of non-organized and spindle-shaped cells exhibiting atypical nuclei,which were divided up by branching vessel and collagen bundles.Immunohistochemical staining showed that this tumor was negative for CD117,CD99,CD68,cytokeratin,calretinin,desmin,epithelial membrane antigen,F8 and S-100,but positive for CD34,bcl-2,α-smooth muscle actin and vimentin.The patient presented no evidence of recurrence during follow-up.SFT arising from abdominal cavity can be diagnosed by histological findings and immunohistochemical markers,especially for CD34 and bcl-2 positive cases.
文摘AIMS To evaluate the sensitivity,specificity and clinical value of hepatic carcinoma associated mem- brane protein antigen (HAg18-1) used as a reagent of serum quick enzyme linked immunosorbent assay (ELISA) for the detection of primary hepatocellular car- cinoma (PHCC). METHODS Serum quick enzyme linked immunosor- bent assay (ELISA) with HAg18-1 as reagent,which was prepared by monoclonal antibody against human hepatic carcinoma,was performed on 100 cases of pri- mary hepatocellular carcinoma (PHCC),5 cases of hepatic biliary carcinoma (HBC),10 cases of metastatic hepatic carcinoma (MHC),20 cases of hep- atitis B (HB),20 cases of liver cirrhosis (LC),20 cas- es of gastrointestinal malignant tumours and 20 cases of gastronintestinal inflammatory diseases (including ulcers). Alpha-fetoprotein (AFP) detection concurrent- ly for each case. Twenty samples of bank blood were tested as controls. RESULTS Positive rate of HAg18-1 ELISA and AFP detection were 81% and 68% in PHCC,20% and 40% in HBC,19% and 20% in MHC,10% and 20% in BH, 10% and 20% in LC,10% and 15% in gastrointestinal malignant tumors,and 5% and 10% in gastrointestinal inflammatory diseases. No positive result of HAg18-1 ELISA and AFP detection occurred in any sample of bank blood. CONCLUSIONS HAg18-1 ELISA has high sensitivity and specificity in the detection of PHCC. HAg18-1 ELISA and AFP detection,if used together,may be complementary to each other in the diagnosis of PHCC.
文摘AIM: To investigate the usefulness of tumor markers and adenosine deaminase in differentiating between tuberculous peritonitis (TBP) and peritoneal carcinoma- tosis (PC). METHODS: A retrospective analysis of data was per- formed on consecutive patients who underwent perito- neoscopic and abdominal computed tomography (CT) evaluations. Among 75 patients at the Seoul National University Hospital from January 2000 to June 2010 who underwent both tests, 27 patients (36.0%) and 25 patients (33.3%) were diagnosed with TBP and PC, re- spectively. Diagnosis was confirmed by peritoneoscopic biopsy. RESULTS: Serum c-reactive protein (7.88:1:6.62 mg/ dL vs 3.12 + 2.69 mg/dL, P = 0.01), ascites adenos- ine deaminase (66.76:1:32.09 IU/L vs 13.89 :l: 8.95 IU/L, P 〈 0.01), ascites lymphocyte proportion (67.77 :1: 23.41% vs 48.36 + 18.78%, P 〈 0.01), and serum- ascites albumin gradient (0.72 + 0.49 g/dL vs 1.05 + 0.50 g/dL, P = 0.03) were significantly different be- tween the two groups. Among tumor markers, serum and ascites carcinoembryonic antigen, serum carbohy- drate antigen 19-9 showed significant difference be- tween two groups. Abdominal CT examinations showed that smooth involvement of the parietal peritoneum was more common in the TBP group (77.8% vs 40.7%) whereas nodular involvement was more common in the PC group (14.8% vs 40.7%, P = 0.04). From receiver operating characteristic (ROC) curves ascites adeno- sines deaminase (ADA) showed better discriminative capability than tumor markers. An ADA cut-off level of 21 IU/L was found to yield the best results of differ- ential diagnosis; sensitivity, specificity, positive predic- tive value, and negative predictive value were 92.0%, 85.0%, 88.5% and 89.5%, respectively. CONCLUSION: Besides clinical and radiologic findings, ascitic fluid ADA measurement is helpful in the differen- tial diagnosis of TBP and PC.
文摘AIM:To compare the outcome of surgical treatment of colorectal adenocarcinoma in elderly and younger patients.METHODS:The outcomes of 122 patients with colorectal adenocarcinoma who underwent surgical treatment between January 2004 and June 2009 were analyzed.The clinicopathological and blood biochemistry data of the younger group(<75 years) and the elderly group (≥75 years) were compared.RESULTS:There were no significant differences between the two groups in operation time,intraoperative blood loss,hospital stay,time to resumption of oral intake,or morbidity.The elderly group had a significantly higher rate of hypertension and cardiovascular disease.The perioperative serum total protein and albumin levels were significantly lower in the elderly than in the younger group.The serum carcinoembryonic antigen level was lower in the elderly than in the younger group,and there was a significant decreasing trend after the operation in the elderly group.CONCLUSION:The short-term outcomes of surgical treatment in elderly patients with colorectal adenocarcinoma were acceptable.Surgical treatment in elderly patients was considered a selectively effective approach.
文摘The data from in vitro and animal experiment study has showed that costimulaory molecule B7 1 plays an important role in antitumor immunity In the present study, B7 1 expression was observed in 130 samples from a veriety of human malignancies by using immunocytochemistry, in situ hybridization and RT PCR combined with dot hybridization and B7 1 specific Mab and probe The results demonstrated B7 1 expression on tumor cells in 76 cases at both protein and mRNA level Forty two specimens were stained with B7 1 HLA ABC and HLA DR Mab and 26 showed that the three antibodies used all were positive Together with the achievement in tumor antigen study, the present findings imply that in most tumors (if not all) the tumor cells have all the requisite element to elicit anti tumor rejection response, the heterogeneous mechanism for tumor escape from immunosurvillance should be emphasized
基金Supported by Grant from the State Key Program of the National Natural Science Foundation of China,No. 81030043
文摘AIM:To evaluate pretreatment serum carcinoembryonic antigen(CEA) as a predictor of survival for patients with locally advanced gastric cancer receiving perioperative chemotherapy.METHODS:We retrospectively studied a cohort of 228 gastric cancer patients who underwent D2 gastrectomy combined with chemotherapy at the Sun Yat-sen University Cancer Center between January 2005 and December 2009.Among them,168 patients received 6-12 cycles of oxaliplatin-based adjuvant(post-operative) chemotherapy,while 60 received perioperative chemotherapy(2 cycles of FOLFOX6 or XELOX before surgery and 4-10 cycles after surgery).Serum CEA was measured using an enzyme immunoassay.The followup lasted until December 2010.RESULTS:In the group that had elevated serum CEA,the difference in survival time between patients receiving perioperative chemotherapy and those receiving adjuvant chemotherapy had no statistical significance(P > 0.05).However,in the group that had normal serum CEA,patients receiving perioperative chemotherapy had a longer survival time.In multivariate analysis,T staging and lymph node metastatic rate were independent prognostic factors for the patients.Perioperative chemotherapy improved the overall survival of patients who had a normal pretreatment CEA level(P = 0.070).CONCLUSION:Normal pretreatment serum CEA is a predictor of survival for patients receiving perioperative chemotherapy.
文摘We have developed and tested chimeric T-cell receptors (TCR) specific for p185HER2. In these experiments, retroviral vectors expressing the N29γ or N29ζ receptors were constructed in pRET6. Amphotropic viral producer cells were established in the GALV-based PG13 packaging cell line. Ficoll purified human peripheral blood lymphocytes (PBL) were virally transduced using an optimized protocol incorporating activation with immobilized anti-CD3/anti-CD28 monoclonal anti- bodies, followed by viral infection in the presence of fibronectin fragment CH296. Transduced cells were co-cultured with human tumor cell lines that overexpress (SK-OV-3) or underexpress (MCF7) p185HER2 to assay for antigen specific im- mune responses. Both CM+ and CD8+ T-cells transduced with the N29γ or N29ζ chTCR demonstrated HER2-specific anti- gen responses, as determined by release of Th1 like cytokines, and cellular cytotoxicity assays. Our results support the fea- sibility of adoptive immunotherapy with genetically modified T-cells expressing a chTCR specific for p185HER2.