Objective: Overexpression of the HER2/neu oncogene is a frequent molecular event in multiple human cancers. Being a cancer antigen, p185 erbB2 is an ideal target for immunotherapy. In order to decrease the immunogenicity of mouse anti-p185 erbB2 monoclonal antibody in human cancer therapy, we constructed the eukaryotic expression vector of anti-p185 erbB2 chimeric monoclonal antibody and verified expression of the chimeric antibody in CHO-dhfr - cell. Methods: The variable regions of light chain and heavy chain were amplified with RT-PCR and inserted into the chimeric antibody vector pWSD2. After CHO-dhfr - cells were transfected with recombination plasmid by lipofectAMINE, the chimeric antibody expressing level was identified with RT-PCR, indirect-ELISA, and Western blot. The specificity of the anti-p185 erbB2 chimeric antibody was testified with ELISA assay and immunoprecipitation. Moreover, the effects of chimeric antibody on the proliferation of breast cancer cell line SKBR3, which is overexpressing p185 erbB2 , were measured with MTT assay in vitro. Results: The anti-p185 erbB2 chimeric antibody eukaryotic expression vector was constructed successfully and the expression of the chimeric antibody in CHO-dhfr - was verified by RT-PCR, indirect-ELISA, and Western blot. ELISA assay showed that chimeric antibody reacted with cells overexpressing p185 erbB2 specifically, but did not react with that non-overexpressing p185 erbB2 . Immunoprecipitation test confirmed that the chimeric antibody could bind to p185 erbB2 specifically. The MTT assay demonstrated that the chimeric antibody could inhibit the growth of SKBR3 cells overexpressing p185 erbB2 . Conclusion: The anti-p185 erbB2 mouse/human chimeric antibody that was expressed in CHO-dhfr - cells can bind to p185 erbB2 specifically and inhibit proliferation of SKBR3 cells overexpressing p185 erbB2 . It has a potential application in biotherapy of cancer.

To produce many epitopes of peptide mut, different tandem repeats containing HER-2 mimetic peptide mut were constructed and expressed. The oligonucleotide coding mut sequence was inserted into pET28a(+) to construct the prokaryotic recombinant plasmid pET28a-mutl. The mut coding sequence was repeatedly inserted into the vector in tandem to obtain a series of plasmids pET28a-mut2, pET28a-mut4, pET28a-mut8 and pET28a-mut16. The plasmids were transformed into E. coli. BL21 and induced by IPTG. The recombinant proteins were expressed in E. coli. The binding analysis verified that the interaction between the tandem peptides mut and Herceptin was increased. In conclusion, the tandem peptides consisted of many antigen epitopes, and laid the foundation for the further study of HER-2 peptide vaccine for biotherapy of cancer with HER-2 overexpression.
Breast Neoplasms ; metabolism ; Cancer Vaccines ; biosynthesis ; immunology ; Epitopes ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression ; Humans ; Peptides ; genetics ; immunology ; metabolism ; Receptor, ErbB-2 ; biosynthesis ; chemistry ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

Objective:To investigate whether increased levels of preoperative carcinoembryonic antigen (CEA) and CA199 were associated with the mortality of patients with advanced colorectal cancer and to determine whether CA199 can be used to discriminate patients with normal preoperative CEA level from good to poor prognosis. Methods:A total of 314 patients with advanced colorectal cancer cases who underwent primary tumor resection were collected from 2014 to 2016 at Peking University Cancer Hospital and Institute. Preoperative CEA and CA199 serum levels were examined using electrochemiluminescence immunoassay. The association of CEA and CA199 with clinicopathologic features and their possible prognostic values were analyzed. Results:In the tested patients, 5.4%of whom had increased CA199 level but not up-regulated CEA level, whereas 28.3%only had increased CEA level, and 21.3%had both CEA and CA199 levels increased. Kaplan-Meier survival curves indicated that patients with elevated preoperative CEA level had worse disease-free survival (DFS) and overall survival (OS) than those with normal CEA (P<0.001 and P<0.001, respectively). Meanwhile, patients with elevated CA199 level had worse DFS and OS than those with normal CA199 (P<0.001 and P<0.001, respectively). Preoperative CA199 level could be used in discriminating patients with normal CEA from good to poor prognosis (P=0.012). Multivariate analysis revealed that elevated CA199 level was an independent prognostic factor for OS in patients with advanced colorectal cancer (HR=2.025, 95%CI=1.331-3.082, P=0.001). Conclusion:Combined detection of preoperative CEA and CA199 can be used in evaluating the prognosis of patients with advanced colorectal cancer. Preoperative serum CA199 level can be used in evaluating the prognosis of patients with colorectal cancer without an increase in CEA level. Patients with increased CA199 level had worse 5-year survival than those with increased CEA level.

OBJECTIVETo explore the predictive value of combination detection of Pgp1 expression in cancer tissue and serum CEA level for the prognosis of colorectal cancer (CRC) patients.

METHODSClinicopathological data, complete 5-year follow-up data and CRC tissue samples of 153 CRC patients with stage I( to II( tumor undergoing radical operation in our department from January 2004 to August 2006 were retrospectively collected. Immunohistochemical staining was used to detect the expression level of Pgp1. The combined evaluation of staining intensity and positive cell percentage was performed to determine the expression level of Pgp1. Pgp1 staining (-) and (+) was defined as low expression; and staining (++) and (+++) as high expression. Electrochemiluminescence immunoassay was used to detect the level of serum CEA. CEA > 5 μg/L was defined as positive. χand Fisher's exact test were performed to analyze the association of Pgp1 expression with CEA level and clinicopathological variables. Moreover, Kaplan-Meier method was used to analyze the survival. Univariate and multivariate Cox proportional hazard regression models were used to evaluate the roles of Pgp1 expression combined with serum CEA level in prognosis prediction.

RESULTSOf 153 patients, 105 were males and 48 females with mean age of 59 (27 to 90) years; 41 cases were rectal cancer, and 112 cases colon cancer; 23 patients were TNM stage I( tumor, and 130 patients stage II( tumor; median follow-up time was 64 months; 30 cases were dead. Positive rate of Pgp1 expression in colorectal cancer tissues was 66.0%(101/153). The expression of Pgp1 was associated with gender, tumor location, and survival during the follow-up (all P<0.05). The preoperative positive rate of serum CEA was 28.1% (43/153). The preoperative serum CEA level was associated with tumor recurrence and survival (all P<0.05). Kaplan-Meier analysis showed the overall 5-year survival rate was 81.7%. The 5-year survival rate of patients with high expression of Pgp1 was 88.1%, which was significantly higher than 69.2% of those with low expression of Pgp1(P=0.003). The 5-year survival rate of patients with preoperative positive serum CEA was 72.1%, which was significantly lower than 86.1% of those with preoperative negative serum CEA(P=0.023). Furthermore, the 5-year survival rate of patients with negative Pgp1 plus positive CEA was 66.7%, which was significantly lower than 91.0% of those with positive Pgp1 plus negative CEA(P=0.002). Univariate analysis showed that gender, Pgp1 expression level, preoperative serum CEA level, and Pgp1 combined with CEA were significantly associated with the prognosis of patients(all P<0.05). Multivariate analysis showed that Pgp1 expression was an independent prognostic factor of CRC [HR(95%CI:1.261 to 64.224), P=0.028].

CONCLUSIONSLow expression of Pgp1 in cancer tissue indicates poor prognosis in patients with stage I( and II( tumor. Combination detection of Pgp1 expression and serum CEA can be applied to predict the prognosis of patients with stage I( and II( colorectal cancer.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; physiology ; Carcinoembryonic Antigen ; blood ; physiology ; Colonic Neoplasms ; physiopathology ; secretion ; Colorectal Neoplasms ; physiopathology ; secretion ; Female ; Fluorescent Antibody Technique ; Humans ; Hyaluronan Receptors ; metabolism ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Proteins ; blood ; physiology ; Neoplasm Recurrence, Local ; physiopathology ; Neoplasm Staging ; Predictive Value of Tests ; Prognosis ; Proportional Hazards Models ; Rectal Neoplasms ; physiopathology ; secretion ; Retrospective Studies ; Sex Factors ; Survival Rate

OBJECTIVETo explore the expression of NF-E1b in colorectal cancer tissues and its association with various clinicopathological parameters and prognosis of the patients.

METHODSClinicopathological and follow-up data of 168 colorectal cancer patients undergoing radical operation at Department of Gastrointestinal Surgery, Peking University Cancer Hospital and Institute from 2005 to 2012 were retrospectively analyzed, including 96 males and 72 females, with mean age of (57.8±11.2) years. The expression of NF-E1b protein was detected in samples of 168 resected colorectal cancer tissues and 45 adjacent non-cancerous tissues by immunohistochemistry. The expression rates of NF-E1b were compared among different clinicopathological features. Moreover, the association between NF-E1b expression and prognosis was analyzed.

RESULTSThe expression of NF-E1b protein located mainly in cytoplasm. Positive rate of NF-E1b expression in adjacent non-cancerous tissues was 17.8% (8/45), which was obviously lower than 67.9%(114/168) of cancer tissues with significant difference (χ(2)=36.376, P=0.000). Clinicopathological parameters analysis suggested that the expression level of NF-E1b in cancer tissues was associated with age (χ(2)=4.862, P=0.030), TNM staging (χ(2)=10.969, P=0.002), lymph node metastasis (χ(2)=7.390, P=0.008) and distal metastasis (χ(2)=17.887, P=0.000). The median follow-up time was 23(1-77) months. The overall 5-year survival of this cohort was 33.3%. Colorectal cancer patients with high levels of NF-E1b expression showed a worse overall survival compared with those with low levels of NF-E1b expression (18.4% vs. 56.6%, P=0.000). Univariate Cox regression analysis showed that tumor location (P=0.034), tumor size (P=0.003), TNM staging (P=0.000), depth of tumor invasion (P=0.009), lymph node metastasis (P=0.000), distant metastasis (P=0.000) and NF-E1b expression level (P=0.001) were associated with the prognosis of colorectal cancer patients. Multivariate Cox regression analysis revealed that tumor diameter >4 cm (HR=2.193,95% CI:1.334 to 3.603, P=0.002), distant metastasis (HR=2.064, 95% CI:1.160 to 3.672, P=0.014) and high NF-E1b expression (HR=1.994,95% CI:1.068 to 3.724, P=0.030) were independent risk factors of predicting poor prognosis of colorectal cancer patients.

CONCLUSIONSNF-E1b expression up-regulates in colorectal cancer tissues. High expression of NF-E1b is associated with poor prognosis of colorectal cancer patients. NF-E1b may serve as a potential target of the treatment for colorectal cancer.

Aged ; Biomarkers, Tumor ; metabolism ; Colorectal Neoplasms ; diagnosis ; metabolism ; Digestive System Surgical Procedures ; Female ; GATA2 Transcription Factor ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Risk Factors ; Up-Regulation

Hsc70-SNCG fusion protein cDNA fragment containing signal peptide sequence of Igkappa, MMP9, and P37 was inserted into the vector pVAX1 to construct recombinant plasmid pVAX-Igkappa-Hsc70-SNCG, pVAX-MMP9-Hsc70-SNCG, and pVAX-P37-Hsc70-SNCG. Three eukaryotic vectors were constructed and verified by restriction enzyme digestion and sequencing. After transfection with recombination plasmids in QM-7 cells, the transient expression and secretion of three fusion proteins were detected by ELISA and Western Blot. The results suggested that Hsc70-SNCG carrying three different signal peptides could be expressed and secreted by transfected cells, and three signal peptides effectively directed secretion of fusion protein by QM-7 cells. BALB/c mice were immunized by three plasmids using gene gun system. The serum levels of anti-SNCG antibodies in mice were measured by ELISA. The results showed that three secreted plasmids could stimulate humoral immune responses to SNCG in mice, which depended on the secreted expression levels induced by signal peptides.
Animals ; Genetic Vectors ; Humans ; Immunoglobulin G ; biosynthesis ; Mice ; Mice, Inbred BALB C ; Neoplasm Proteins ; biosynthesis ; genetics ; immunology ; Plasmids ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Transfection ; gamma-Synuclein ; biosynthesis ; genetics ; immunology

Objective The radioactivity level survey was carried out in Weizhou Island area of Beihai, to comprehensively master the radiation environment status in the area, understand the natural radioactivity level and its distribution law, and provide basic data for scientific evaluation of radiation environment quality. Methods According to the relevant standards and technical specifications, the air absorbed dose rate of gamma radiation in Weizhou Island area was monitored from April 2020 to March 2021, the concentrations of radionuclides in water, soil and other environmental samples were sampled and analyzed, and the monitoring results were analyzed and discussed in combination with the regional characteristics. Results The results show that the air absorbed dose rate of gamma radiation in Weizhou Island area ranges from 0.2～122 nGy/h; the activity concentrations of ²³⁸U, ²²⁶Ra, ²³²Th, ⁴⁰K and ¹³⁷Cs in solid samples range 17.0～37.3 Bq/kg, < 0.501～29.5 Bq/kg, < 0.766～54.3 Bq/kg, 18.7～369 Bq/kg and < 0.212～1.48 Bq/kg, respectively, the concentrations of U and Th in well water and reservoir water range 0.065～0.25 μg/L, 0.046～0.079 μg/L, the activities of ²²⁶Ra, ⁴⁰K, total α and total β range 1.42～3.08 mBq/L, 0.069～0.231 Bq/L, 0.025～0.163 Bq/L, 0.082～0.572 Bq/L, respectively, the concentrations of U and Th in seawater samples range 1.81～2.25 μg/L, 0.634～0.648 μg/L, and the activities of ²²⁶Ra, ⁴⁰K, ⁹⁰Sr and ¹³⁷Cs range 9.38～19.7 mBq/L, 11.3～11.8 Bq/L、0.193～0.866 mBq/L、1.13～1.42 mBq/L. Conclusion The environmental ionizing radiation level in Weizhou island of Beihai is in the range of background fluctuation and at a relatively low level, indicating that the radiation environmental quality of Weizhou Island and its surrounding areas is good.

OBJECTIVETo investigate the predictive value of preoperative detection of CEA and CA199 for prognosis in patients with stage II-III colorectal cancer (CRC).

METHODSClinicopathological and follow-up data of 266 patients with stage II-III colorectal cancer confirmed by pathology and undergoing radical resection in our department from 2004 to 2006 were retrospectively analyzed. The 5-year overall survival (OS) and disease-free survival (DFS) between normal CEA patients and increased CEA ones, and normal CA199 patients and increased CA199 ones were compared respectively. Moreover, the risk factors of OS and DFS were examined.

RESULTSAmong these 266 patients, 119 (44.7%) had preoperative elevated CEA, and 74 (27.8%) had increased CA199. The median follow-up time was 63 months. Kaplan-Meier survival curves indicated that patients with elevated preoperative CEA had worse 5-year OS (54.6% vs. 72.8%, P=0.001) and DFS (75.2% vs. 83.3%, P=0.042) than those with normal CEA respectively. Meanwhile, patients with elevated CA199 had worse OS (45.9% vs. 71.9%, P=0.000) and DFS (74.2% vs. 81.8%, P=0.047) than those with normal CA199 respectively. Multivariate analysis revealed that male (HR=3.016, 95% CI: 1.536-5.919, P=0.001), lymph node metastasis (HR=2.278, 95% CI: 1.272-4.081, P=0.006), and preoperative elevated CEA (HR=1.794, 95%CI: 1.022-3.149, P=0.042) were independent prognostic factors of DFS. While vascular thrombosis (HR=2.041, 95% CI: 1.294-3.221, P=0.002), lymph node metastasis (HR=2.480, 95% CI:1.548-3.972, P=0.000), and preoperative elevated CA199 (HR=2.145, 95% CI:1.414-3.254, P=0.000) were independent prognostic factors of OS in patients with stage II-III CRC.

CONCLUSIONCombined detection of preoperative CEA and CA199 can be used in evaluating the prognosis of patients with stage II-III CRC.

Antigens, Tumor-Associated, Carbohydrate ; blood ; Carcinoembryonic Antigen ; blood ; Colorectal Neoplasms ; diagnosis ; Disease-Free Survival ; Female ; Humans ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Male ; Multivariate Analysis ; Neoplasm Staging ; Predictive Value of Tests ; Prognosis ; Retrospective Studies ; Risk Factors ; Survival Rate

Objective To investigate the predictive value of preoperative detection of CEA and CA199 for prognosis in patients with stage Ⅱ-Ⅲ colorectal cancer (CRC). Methods Clinicopathological and follow-up data of 266 patients with stage Ⅱ-Ⅲ colorectal cancer confirmed by pathology and undergoing radical resection in our department from 2004 to 2006 were retrospectively analyzed. The 5-year overall survival(OS) and disease-free survival(DFS) between normal CEA patients and increased CEA ones, and normal CA199 patients and increased CA199 ones were compared respectively. Moreover, the risk factors of OS and DFS were examined. Results Among these 266 patients, 119 (44.7%) had preoperative elevated CEA, and 74 (27.8%) had increased CA199. The median follow-up time was 63 months. Kaplan-Meier survival curves indicated that patients with elevated preoperative CEA had worse 5-year overall survival (OS) (54.6% vs. 72.8%, P=0.001) and disease-free survival (DFS)(75.2% vs. 83.3%, P=0.042) than those with normal CEA respectively. Meanwhile, patients with elevated CA199 had worse OS(45.9% vs. 71.9%, P=0.000) and DFS (74.2% vs. 81.8%, P=0.047) than those with normal CA199 respectively. Multivariate analysis revealed that male (HR=3.016, 95% CI:1.536-5.919, P=0.001), lymph node metastasis (HR=2.278, 95% CI:1.272-4.081, P=0.006), and preoperative elevated CEA (HR =1.794, 95% CI:1.022-3.149, P =0.042) were independent prognostic factors of DFS. While vascular thrombosis (HR=2.041, 95% CI:1.294-3.221, P=0.002), lymph node metastasis (HR=2.480, 95% CI:1.548-3.972, P=0.000), and preoperative elevated CA199 (HR=2.145, 95% CI:1.414-3.254, P=0.000) were independent prognostic factors of OS in patients with stage Ⅱ-Ⅲ CRC. Conclusion Combined detection of preoperative CEA and CA199 can be used in evaluating the prognosis of patients with stage Ⅱ-Ⅲ CRC.

Objective To investigate the predictive value of preoperative detection of CEA and CA199 for prognosis in patients with stage Ⅱ-Ⅲ colorectal cancer (CRC). Methods Clinicopathological and follow-up data of 266 patients with stage Ⅱ-Ⅲ colorectal cancer confirmed by pathology and undergoing radical resection in our department from 2004 to 2006 were retrospectively analyzed. The 5-year overall survival(OS) and disease-free survival(DFS) between normal CEA patients and increased CEA ones, and normal CA199 patients and increased CA199 ones were compared respectively. Moreover, the risk factors of OS and DFS were examined. Results Among these 266 patients, 119 (44.7%) had preoperative elevated CEA, and 74 (27.8%) had increased CA199. The median follow-up time was 63 months. Kaplan-Meier survival curves indicated that patients with elevated preoperative CEA had worse 5-year overall survival (OS) (54.6% vs. 72.8%, P=0.001) and disease-free survival (DFS)(75.2% vs. 83.3%, P=0.042) than those with normal CEA respectively. Meanwhile, patients with elevated CA199 had worse OS(45.9% vs. 71.9%, P=0.000) and DFS (74.2% vs. 81.8%, P=0.047) than those with normal CA199 respectively. Multivariate analysis revealed that male (HR=3.016, 95% CI:1.536-5.919, P=0.001), lymph node metastasis (HR=2.278, 95% CI:1.272-4.081, P=0.006), and preoperative elevated CEA (HR =1.794, 95% CI:1.022-3.149, P =0.042) were independent prognostic factors of DFS. While vascular thrombosis (HR=2.041, 95% CI:1.294-3.221, P=0.002), lymph node metastasis (HR=2.480, 95% CI:1.548-3.972, P=0.000), and preoperative elevated CA199 (HR=2.145, 95% CI:1.414-3.254, P=0.000) were independent prognostic factors of OS in patients with stage Ⅱ-Ⅲ CRC. Conclusion Combined detection of preoperative CEA and CA199 can be used in evaluating the prognosis of patients with stage Ⅱ-Ⅲ CRC.