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Esophageal squamous cell carcinoma (ESCC)is one of the most predominant malignancies worldwide. ESCC develops gradually from the precancerous lesions. The researches about biomarkers mainly concentrate in the nucleic acid and protein,including cytogenetic abnormalities,amplifications,deletions,muta-tions,breakage and fusions of genomic DNA,expression changes of mRNA,microRNA (miRNA)and long non-coding RNA (lncRNA)in RNA level,and expression level,subcellular localization,and post-translational modi-fications of proteins. Studies on the molecular alterations in precancerous lesions and early cancer of the esopha-gus will contribute to the exploration of biomarkers for early diagnosis and intervention targets for this disease.
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Abnormal genomic DNA methylation is an important epigenetic change in malignant tumors. Esophageal squamous cell cancer is one of common malignant tumors in our country. In this paper summarized and discussed the progress of genomic DNA methylation in the esophageal squamouscell cancer, including the level of genomic DNA methylation, frequent abnormally methylated genes, methylation markers and potential targets, etc. This paper might provide candidate biomarkers and targets for further studies on the mechanism of the tumorigenesis and development of the esophagealsquamouscell cancer, as well as for the clinical application of esophageal cancer.
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OBJECTIVE:To provide a reference for promoting and strengthening pharmaceutical services by retail pharmacies in rural area in Yongzhou of Hunan. METHODS:Investigation was conducted on 13 aspects including the service concept,service mode,service process,service contents,hardwares and softwares and professional quality of practitioners of 357 retail pharmacies in rural area in Yongzhou of Hunan to know about current pharmaceutical services. RESULTS & CONCLUSIONS:In this area, 78.9% of pharmacies operated in the name of a licensed pharmacist or pharmacist who did not work in that pharmacy;only 53.3%of practitioners were pharmacy graduates;the proportion of the practitioners who takes part in training every year was less than 30.0%,average training time was not more than 8 hours;65.3% of practitioners in the pharmacies were not aware of the concept of pharmaceutical services;the pharmacies which were not equipped with basic hardwares and softwares for pharmaceutical servic-es accounted for 47.0%,those which had not established a pharmaceutical service management process accounted for 87.4%;and less than 10.0% of retail pharmacies provided door-to-door pharmaceutical services. Safe,effective and rational drug use should be achieved by intensifying the management and training for retail pharmacy practitioners,standardizing the contents of pharmaceutical services,establishing standard pharmaceutical service process,defining basic hardwares and softwares for pharmaceutical services, strengthening performance appraisal and evaluation for pharmaceutical services and urging the pharmacy to serve society.
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<p><b>OBJECTIVE</b>The aim of this study was to find out the value of chromosome aneuploidy in early diagnosis and prediction of postoperative recurrence of gastric adenocarcinoma (GAC).</p><p><b>METHODS</b>Tissue samples, including 49 GAC, pairing pericancerous mucosa and normal gastric mucosa from the distant cutting margin were use in this study. Two centromere probes, Cen11 and Cen20 specific for chromosomes 11 and 20 were analyzed by FISH . The clinicopathological data were summarized.</p><p><b>RESULTS</b>The incidence of aneuploidy of chromosome 11 in the tumors, pericancerous mucosa and normal gastric mucosa from the distant cutting margin were 83.7%, 40.8%, and 16.3%, respectively (P < 0.001), and those of chromosome 20 were 87.8%, 53.1%, and 16.3%, respectively (P < 0.001). The aneuploidy of Cen 11 displayed a significant correlation with Lauren's claasification, and lymph node metastasis (P < 0.05 for both). The pericancerous mucosa and normal gastric mucosa from the distant cutting margin displayed mainly chronic inflammatory changes, intestinal metaplasia and dysplasia.</p><p><b>CONCLUSIONS</b>Aneuploidy of Cen11 and Cen20 in pericancerous mucosa may be used as a candidate marker for early diagnosis of gastric adenocarcinoma and may have a predictive value of postoperative recurrence.</p>
Subject(s)
Humans , Adenocarcinoma , Diagnosis , Genetics , Aneuploidy , Chromosomes, Human, Pair 11 , Gastric Mucosa , Lymphatic Metastasis , Neoplasm Recurrence, Local , Diagnosis , Genetics , Stomach Neoplasms , Diagnosis , GeneticsABSTRACT
To rapidly select potent anti-VSTM1-v2 scFv (single-chain antibody fragment) by construction and screening of a humanized scFv library in which a murine VH-CDR3 library was grafted onto a human scFv framework. A murine VH-CDR3 library was amplified from anti-VSTM1-v2 murine cDNA and grafted on human scFv (VH3-VK1) framework. Anti-VSTM1-v2 scFv templates were selected and enriched through ribosome display, TA-cloned into expression vector, and transformed into BL21 (DE3) for soluble expression of target scFv. A total of 1000 clones were randomly picked. Positive ones were first identified using colony PCR, indirect ELISA, Western blotting and then verified with sequencing and dose response ELISA. At last an anti-VSTM1-v2 humanized scFv with good binding affinity (EC50 = 21.35 nmol x L(-1)) was selected from the humanized library of 10(12) members generated in this study. This scFv antibody might have potential applications. This study provides a new approach for rapid screening of humanized antibodies.
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Since the publication of a high-throughput DNA sequencing technology based on PCR reaction was carried out in oil emulsions in 2005, high-throughput DNA sequencing platforms have been evolved to a robust technology in sequencing genomes and diverse DNA libraries. Antibody libraries with vast numbers of members currently serve as a foundation of discovering novel antibody drugs, and high-throughput DNA sequencing technology makes it possible to rapidly identify functional antibody variants with desired properties. Herein we present a review of current applications of high-throughput DNA sequencing technology in the analysis of antibody library diversity, sequencing of CDR3 regions, identification of potent antibodies based on sequence frequency, discovery of functional genes, and combination with various display technologies, so as to provide an alternative approach of discovery and development of antibody drugs.
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Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.
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Objective To investigate the role of Kit signal pathway in the proliferation and spontaneous rhythmic motility of interstitial cells(ICs)in the bladder of guinea pigs.Methods Fourteen Guinea pigs aged 45-60 days were administered with Imatinib intragastrically for 20 and 40 days.Three were fed on normal saline as control.The morph and number of ICs were observed by immunofluorescence,Kit protein expression was detected by Western blot and spontaneous rhythmic motility of bladder were investigated in in vitro organ bath.Results Quite a few ICs were observed in the bladder of controls.After administration of Imatinib,the amount of ICs and the expression of Kit protein decreased gradually,and the spontaneous rhythmic contraction weakened.Conclusion ICs might be the pacemaker of bladder in guinea pigs.Kit protein might play an important role in the survival and functional maintenance of ICs.
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Objective To investigate the microsatellite instability(MSI) and monomorphism at microsatellite DNA BAT 26 locus in esophageal mucosa. Methods Genomic DNA extracted from tissues was amplified by PCR. PCR products were run on 9% denaturing polyacrylamidegel and stained with silver. Then MSI and monomorphism of microsatellite at BAT 26 locus in normal tissues at the incised edge and esophageal cancer tissues in 45 cases of esophageal caner were analyzed. Results All normal specimens showed no change in the length of MS DNA at BAT 26 locus. MSI was detected in 3 out of 45(6 7%) cases of esophageal cancer. Conclusion BAT 26 has complete monomorphism in genome of normal esophageal tissues, but is not sensitive to the identification of MSI.
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Objective To detect the state of microsatellite instability (MSI) and investigate the relationship between MSI and clinical pathological parameters in esophageal cancer. Methods MSI was analyzed by PCR SSCP method. Results MSI was detected (22 22%) in 45 cases of esophageal cancer. Effects of MSI on clinical stage, pathological classification, lymph node metastasis, and invasion were not found. Conclusion MSI may be an early molecular pathway and play a certain role in the development of the esophageal cancer.
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Objective To summarize the experience of surgical treatment of ascending aortic aneurysms and aortic dissections. Methods From February 2001 to October 2005, 31 patients including 26 male, 5 female, aged 41.3 years old (range 14-72) received surgical management. Twenty cases were diagnosed as ascending aortic aneurysm and aortic root aneurysm, 8 as Standford A dissection, 3 as Stanford B dissection. Twenty-one patients underwent classic Bentall procedure in which VSD repair was carried out in 1 case, mitral valvoplasty in 2 and mitral valve replacement in 2; Four patients underwent modified Bentall procedure (coronary button technique); Three patients underwent Wheat procedure; The remaining 3 patients with Stanford B dissection underwent graft replacement of descending aorta. Results There was no death during hospital stay that lasted 13-46 d with an average of 16.4 d after operation. The mean clinical follow-up was (21?18.5) months (range 1-63 months). One patient died without describable cause two years later. One patient had ascending aorta-pulmonary artery fistula at color Doppler examination half a year later. One patient was detected rupture of distal anastomoses half a year after operation and underwent stent-graft, SG. Conclusion The surgical treatment of aortic aneurysms and aortic dissections could be carried out safely based on the accurate diagnosis, specific surgical strategy and fine technique.
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<p><b>OBJECTIVE</b>To establish a long-standing cell line of esophageal squamous cell carcinoma (ESCC) in pursuit of a model for in vitro study of carcinogenesis.</p><p><b>METHODS</b>Small tissue blocks taken from resected specimens of esophageal cancer were cultured, and cell line EC9706 was established. The biologic properties of EC9706 were characterized. Comparative genomic hybridization(CGH) was performed on the cell line.</p><p><b>RESULTS</b>The growth curve of EC9706 was detected. The cell generation time was 26 hours. The plate colony forming efficiency is 91.9%, with the capacity of forming clones in soft agar. EC9706 cells show high tumorigenecity as indicated by the rapid regeneration of moderate-poor-differentiated squamous cell carcinomas after injection into nude mice. CGH analysis indicated copy number gains of 1p1, 1q2-4, 2p1, 2q1, 5p, 7p14, 7q21, 11q1, 15q2, 20q and losses of 2p2, 2q2, 3p, 4, 9p, 14, 18, Xq. High-level gain of 5p was observed.</p><p><b>CONCLUSION</b>Established cell line EC9706 can serve as a useful tool for studying the carcinogenesis of ESCC.</p>
Subject(s)
Aged , Animals , Humans , Male , Mice , Carcinoma, Squamous Cell , Genetics , Pathology , Cell Division , Chromosome Aberrations , Esophageal Neoplasms , Genetics , Pathology , Mice, Nude , Neoplasm Transplantation , Nucleic Acid Hybridization , Methods , Transplantation, Heterologous , Tumor Cells, CulturedABSTRACT
Objective To investigate the role of suramin in inhibiting the angiogenesis of lung cancer. Methods The protei n expression of VEGF and its receptor fms-like tyrosine kinase(Flt), kinase ins ert domain-containing receptor(KDR) in lung cancer cell A549 and ECV-304 was studied with immunohistochemical technique. The anti-angiogenesis effect of su ramin was evaluated with immunohistochemical technique and cell culture. Results Suramin showed no effect on the production of lung cancer cel ls and their secretion of VEGF. No possitive expression of Flt, KDR in A549 was found in the suramin interfered group and non-interfered group, but VEGF w as e xpressed in both groups. The Flt and KDR expressions in ECV-304 cells were decr eased significantly after the treatment of suramin at 0.25 ng/ml and 2.5 ng/ml r espectively. There was no effect on proliferetion of A549 and ECV-304 after sur amin in terference. Conclusion The mechanism of inhibiting-angiogenes is of suramin is pro bably due to reducing the VEGF receptor expression in vascular endothelial cells and inhibiting the binding of VEGF and its receptors.
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Objective Coronary artery bypass grafting (CABG)i n 13 patients was analyzed. Methods 9 patients were performed C ABG with cardiopulmonary bypass, 4 patients undergone off-pump coronary artery bypass(OPCAB). Among the 4 patients, 3 undergone transmyocardial laser revascula rization concomitantly. 2 patients with single-vessel disease, 3 with double-v essel disease, 7 with triple-vessel disease and 1 with left main coronary arter y disease. The average bypass per patient was 2.3. Results All patients survived, 11 patients were angina free, 2 were angina relief. C onclusion CABG is a safe operation, OPCAB may reduce blood transfusion and complication, patients recover more quickly after OPCAB compared with those with CABG.
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Objective To study the status of cystatin B gene expression in human esophageal cancer. Methods Expression of cystatin B gene was detected by RT PCR in 41 pairs of human esophageal cancer tissues, matched adjacent normal mucosa, human esophageal cancer cell line EC9706 and human lung cancer cell line GLC82. Results cystatin B gene expression was down regulated in 82.9% (34/41) of human esophageal cancer tissues while expressed at high level in all matched adjacent normal esophageal mucosa, and was significantly corelated to lymph node metastasis ( P
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Objective To investigate the mechanisms of Mal gene down regulated in human esophageal cancer. Methods We amplified the 5′regulatory region of Mal gene in normal placenta tissue and human esophageal cancer cell lines EC9706、EC8712、EC109 by optimization the PCR methods, cloned and sequenced the PCR products and compare the sequence. Results Human esophageal cancer cell lines EC9706、EC8712、EC109 all have the 654 th of 5′regulatory region of Mal gene T to C changes, corresponding codon changed from TCC to CCC, and the amino acid which it codes changed from serine to proline; EC9706 also has the 657 th of 5′regulatory region of Mal gene T to C changes, corresponding codon changed from TGC to CGC, the amino acid which it codes changed from cysteine to arginine; EC8712 has the 139 th of 5′regulatory region of Mal gene A to G changed, corresponding codon changed from GGA to GGG, the amino acid which it codes remains glycine. Conclusion Our results suggested there are may point mutation in 5′ region of Mal gene in human esophageal cancer.