[show abstract][hide abstract] ABSTRACT: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer.
Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer. RT-PCR was used to amplify the heavy chain Fd and light chain kappa and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected.
The amplified fragments of Fd and kappa gained by RT-PCR were about 650 bp. Fd and kappa PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 x 10(6). The libraries were enriched about 120-fold by 3 cycles of adsorption-elution-multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5 clones of Fab phage antibodies which had binding activities with antigens of colorectal cancer.
The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer.
World Journal of Gastroenterology 01/2002; 7(6):811-5. · 2.55 Impact Factor
[show abstract][hide abstract] ABSTRACT: Both SPA immunological fecal occult blood test (SPA FOBT test) and detection of T-antigen in rectal mucus (Shams' test) were used as screening tests in asymptomatic mass screening to evaluate the complementary effect of both tests for detection of colorectal cancers. SPA FOBT test showed a positive rate of 11.1% and shams' test 8.9% among 7,740 subjects. In 498 cases with positive screening test, 11 cases of carcinomas and 88 adenomas were found with colonoscopy. Only 9 cancers and 55 adenomas showed positive result in SPA FOBT test and 8 cancers and 51 adenomas in Shams' test. Both tests combined could enhance the detective rate of cancer in asymptomatic mass screening from 81.8% with SPA FOBT test or 72.7% with Shams' test to 90.9%. This complementary effect was more obvious in adenoma detection. It is suggested that there were some shortcomings in sequential FOBT test for cancer detection, the combined use of the two different screening tests for detection of colorectal cancer could decrease the rate of missed detection.
Zhonghua nei ke za zhi [Chinese journal of internal medicine] 07/1994; 33(6):367-9.
[show abstract][hide abstract] ABSTRACT: The sugar moiety detected from rectal mucus by the Galactose oxidase-schiff (Shams' test) is considered a substitutive test for immunological fecal occult blood test (FOBT) in screening colorectal carcinoma. Two strategies of screening were applied in 6480 subjects over 40 years of age, and 130 cm flexible colonoscope used for sigmoidoscopy or pancolonoscopy. Of them, 3820 were taken for immune FOBT (SPA test) and Shams' test. Only those who showed positive tests were chosen for 60 cm flexible sigmoidoscopy, while another 2660 subjects for both sigmoidoscopy and tests at the same time. Additionally, 130 cm flexible pancolonoscopy was carried out in 103 individuals with positive Shams' test for evaluating the false positive rate. Shams' test showed a sensitivity of 85.7% for colorectal cancer, 47.1% for adenomas in preselected patients, while the positive rate of SPA test were 90.5% and 41.2% respectively. In 3820 asymptomatic subjects undergoing sequential screening (aged 45 years and higher), Shams' test showed 9.1% positive, SPA showed 11.2% and 620 (16.2%) subjects were selected for sigmoidoscopy based on their positive galactose oxidase result or positive FOBT result. Two early stage carcinomas and 33 adenomas (0.32% and 4.2% respectively in sigmoidoscopy) were found. Another 2 660 subjects were taken for sigmoidoscopy screening. Four carcinomas and 78 adenomas were found. Of them, only two carcinoma (50%) and 17 (21.8%) or 22 (28.2%) adenomas were positive in Shams' or SPA test. But both tests combined in screening showed a rate of 61.3% in adenomas and 75.0% in cancers. 103 subjects with positive Shams' test were taken for pancolonoscopy. 82.5% showed no lesions.(ABSTRACT TRUNCATED AT 250 WORDS)
Chinese medical journal 11/1993; 106(10):739-42. · 0.90 Impact Factor
[show abstract][hide abstract] ABSTRACT: It has been reported that curcumin (diferuloylmethane) could inhibit growth of several types of malignant cells both in vitro and in vivo. However, the mechanism of its action is unknown. In this study, we investigated the inhibitory effects of curcumin on human colon carcinoma cell (Lovo) growth and its mechanism of action in vitro by means of growth assay, colony formation assay, MTT assay, cell cycle and apoptosis analysis. Curcumin inhibited cell growth in a dose-dependent manner. The ability of Lovo cells treated with curcumin to form colonies was depressed. MTT test showed that curcumin was cytotoxic to cells. Lovo cells treated with curcumin were largely accumulated in S, G2/M phase which prevented cells from entering the next cell cycle. Apoptosis induced by curcumin was confirmed by characteristic ladders and cellular morphological changes. Curcumin can inhibit Lovo cells growth and the cellular mechanism responsible for the action is to arrest the cell cycle in S, G2/M phase and to induce apoptotic cell death.
Anticancer research 19(5A):3675-80. · 1.71 Impact Factor
[show abstract][hide abstract] ABSTRACT: Helicobacter pylori infection is the major etiological factor of chronic active gastritis and most peptic ulcer disease, and is closely associated with gastric cancers such as adenocarcinoma and MALT lymphoma. Since the Hp adhesin conservative region (AB) is outer membrane protein and porin type component, while these two kinds of protein are the excellent immunogen candidates of vaccination. The gene ab was amplified by PCR and inserted into the prokaryotie expression vector pET-22b(+) and expressed in the BL21 (DE3) E. coli strain. DNA sequenced showed one open reading frame of 588 bp, which encoded polypeptides of 195 amino acids. SDS-PAGE and scan analysis show the AB molecular mass is 22.5 ku and recombinant protein amounted to 29% of the total bacterial protein. The AB purity amounted to 96% through affinity chromatography. Western blot analysis of AB confirmed it could be specially recognized by serum from rabbit immunized with AlpA. Acquire of AB established foundation for further studying the molecular adhesin, prevention and cure immunity mechanism of the adhesin.
Progress in Biochemistry and Biophysics, v.29, 922-926 (2002).
[show abstract][hide abstract] ABSTRACT: AIM:To investigate effect of cytokines in the immunopathogenesis of Helicobacter pylori (H. pylori). METHODS:Forty-eight patients (37 with chronic gastritis and 11 with duodenal ulcer (DU), and aged 21-63 yrs) were biopsied under endoscopy for gastric mucosa culture in vitro and H. pylori detection. Rapid urease test, H. pylori culture and Warthin-Starry silver stains were used for H. pylori diagnosis. H. pylori was defined as positive when any 2 of these tests were positive. Each of 3 antrum pylori biopsies was cultured in vitro for 24 h by adding 1mL RPMI 1640 (37℃, 50mL·L -1 CO2). Supernatants were centrifuged and used for the detection of interleukin (IL) -6, IL-8 and tumor necrosis factor-a (TNF-α ) by ELISA. Total proteins of the biopsies were assayed by a modified Lowry method. An- other 8 cases were cultured with RPMI 1640 added by H. pylori vacuolating cytotoxin (VacA). The results were ex- pressed as ng orµg per gram protein (ng·g -1 or µg·g -1 ).