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Objective To detect the expression of human telomerase reverse transcriptase (hTERT ) and telomerase in hu‐man fetal gastric fibroblasts .Methods Human fetal gastric fibroblasts were isolated and cultured ,epithelial cells were exduded by CK‐18 immune staining .hTERT protein was determined by indirect immunofluorescence .Telomerase activity was analyzed using telomeric repeat amplification protocol assay (TRAP)while the same analysis in adult gastric fibroblast ,which as to be positive con‐trol .Results Cultured fibroblasts were CK‐18 negative .The positive hTERT immunostaining was detected in both cellular cyto‐plasm and nuclear compartments .Amplified telomeric repeats (about 170 bp) in human fetal gastric fibroblasts were longer than those (160 bp) in adult gastric fibroblasts .Conclusion hTERT and telomerase were expressed in human fetal gastric fibroblasts .
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Objective Colorectal polyp is a precancerous lesion of colorectal cancer .Aim of the study was to explore the risk fac-tors of colorectal polyp malignant transformation .Methods The related information of 75 084 colonoscopies performed from 2003 to 2012 in Southwest Hospital in Chongqing were collected and the relationship between polyp malignant transformation and the pa-tient age ,sex ,polyp location ,size or histological types was analyzed .Results From 2003 to 2012 ,polyps were diagnosed in 14 806 cases of the total 75 084 patients with a 19 .72% detection rate .There were significant difference of the left-side and right-side pol-yp detection rate in different age groups ,and the frequency of polyps distributed in the whole colorectum increased with the increase of age .The rates of epithelial neoplasia and malignant transformation increased with age .At the same time ,malignant transforma-tion rate was significant higher in polyps located in left-side than that in right-side (P<0 .0167) ,in adenoma than that in inflamma-tory hyperplastic polyp (P<0 .01) .The larger diameter and the more villus ,the higher rate of malignant transformation .Conclusion Patient age ,polyp size ,location and histological type could be considered as the significant predictors of colorectal polyp malignant transformation .It may be useful to treat the polyp with endoscopy in patient with age more than 45 and adenoma whose diameter was not less than 1 cm ,located in left-side for prevention of colorectal cancer .
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Advanced studying doctors play important roles in the clinical services, and how to train them to improve training quality is worth investigating. We classified them into three types such as the clinical skills-improved, the special skills-trained and clinical knowledge eextensively-spread, then employed the individual teaching methods and emphasized the medical ethics during the training, which is not only beneficial to us, but also of great importance and necessity to advanced studying doctors themselves.
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Objective To detect the differential expression genes(DEGs)between Barrettg esophagus(BE)and normal esophagus with oligomicroarray,and to explore the target genes related to the development of BE.Methods The total RNAs of matched BE and normal esophagus mucosa from saIne patient were isolated with one step Trizol method.Matched RNAs were qualified with 10g/L agarose gel electrophoresis.After tRNA purification,cRNAs were synthesized and labeled with fluorescence.which were tIlen hybridized with Agilent oligomicroarray containing 30,968 probes.The fluorescence intensity features were detected by Agilent scanner and quantified by software Feature Extraction.Results On average,2 biopsies by disposable jumbo biopsy forceps provided approximately 5μg RNA required for microarray.The total RNA,reverse transcription product and fluorescence labeled cRNA were all of high quality.Among 2-fold DEGs,there were 142 up-regulated genes and 284 down-regulated genes including 15 bel-2 related genes such as bel-2,MCL1,BAX,BIK and BCLAF1 Conclusion Microarray-based studies are feasible in endoscopically obtained tissues.The development of BE is a complicated process involving multi-genes,in which abnormal expression of bel-2 family related genes might be involved,but the exact mechanism needs further research.
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Objective To evaluate the differences, including clinical symptoms, endoscopic and histopathologic findings, status of Helicobacter pylori (Hp) infection and cytokeratin (CK) expressions, be-tween Barrett esophagus (BE) and heterotopic gastric mucosa (HGM) in upper esophagus. Methods Clinical data of 152 patients with BE and 52 patients with HGM in upper esophagus diagnosed from February 2004 to September 2005 were retrospectively studied. The parameters being compared include-ed clinical manifestations, conventional and magnifying endoscopic findings, histopathological findings, Hp infection determined by rapid urease test and Warthin-Starry staining and expression of CK phenotypes detec-ted by immunohistochemistry. Results Gastro-esophngeal reflux was observed in 64. 5% of patients with BE (98/152), higher than that in patients with HGM ( 13.5%, 7/52, χ2 = 40. 36, P < 0. 01 ). Endoscopic faveolus of BE mucosa included 46 cases of spot pattern, 65 striations and 41 villiform patterns, while those of HGM were all striation patterns. The histologic classification in BE included 56 cases of fundic type, 39 junction type and 57 specialized intestinal metaplasia, while in HGM mucesa, 31 cases of fundic type, 16 junction type and 5 antrum type were diagnosed, and no goblet cells were found. Moderate and severe infil-tration of inflammatory cells in BE mucosa was 63.2% (96/152), which was significantly higher than that in HGM mucosa (15/52, 28. 8%, P<0. 01). However, no difference was found in gastric antrum inflam-mation between the two groups (44.7%, 68/152, vs. 51.9%, 27/52, P>0.05). No difference was ob-served in prevalence of Hp infection between BE and HGM groups (P >0. 05 ), either in involved mucosa or in antrum. CK7 was not expressed in HGM or normal squamons mucosa, but was expressed in BE. CK20 and CK19 were expressed in both HGM and BE, and CK13 expression was found in some BE nmcosa including gas-tric metaplesia (55/95) and intestinal metaplasia (29/57) but not in HGM mucosa. Conclusion There are differences between HGM and BE, in regarding of reflux symptoms, magnifying endoscopic findings, histo-logical types and CKs expressions, which may be indicators to make differential diagnosis.
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Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.
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Objective To investigate the relationship between single-nucleotide polymorphisms of IVS13-98G/T of human protection of telomeres 1 (hPOT1) genes and gastric cancer. Methods A total of 168 patients with gastric cancer (gastric cancer group) who had been admitted to Wuwei Cancer Hospital, Wuwei People's Hospital and Liangzhou District Hospital from December 2005 to July 2006 and 156 healthy people (control group) were genotyped by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) method, and the relationship between the distribution of genotypes and the risk faetors of gastric cancer was analyzed. The distribution of genotypes and allele frequency between the 2 groups were compared by chi-square test. Hardy-weinberg equilibrium test was adopted to determine whether the distribution of genotypes and allele frequency were representative. Relative risk and 95% confidence interval were calculated by non-conditional Logistic regression model. Results The frequencies of genotypes GG, GT and TT of hPOT1IVS13-98 G/T were 21.4%, 41.7% and 36.9% in gastric cancer group, and 24.4%, 51.9% and 23.7% in control group. The allele frequencies of G- and T-allele were 42.3%, 57.7% in gastric cancer group, and 49.7%, 50.3% in control group. There was no significant difference in allele frequencies of G- and T-allele between the 2 groups (X~2=3.58, P>0.05). Compared with genotype TT, the relative risks for GT and GG were 0.439 (95% CI: 0.251-0.767, P < 0.05) and 0.514 (95 % CI: 0.264-0.999, P=0.05). There was no influence of different genotypes of hPOT1IVS13-98 G/T, sex, smoking history, family history of cancer on gastric cancer. Conclusion Single-nucleotide polymorphisms of IVS13-98G/T of hPOT1 may be a protective factor of gastric cancer.
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Objective To investigate the effect of rebamipide on chronic non-atrophic gastritis (NAG) with erosion and its protection of gastric mucosa from Helicobacter priori(Hp) associated lesions.Methods Patients(n=452)with endoscopically confirmed NAG with erosion from 11 hospitals in China were enrolled and randomly assigned at a ratio of 3:1 to receive either rebamipide(100 mg t.i.d.)or sucralfate(1.0 t.i.d.)for 8 weeks.Hp infected patients received eradication treatment before randomization.Symptoms,endoscopic scores and histological changes were recorded before and after therapy.Concentrations of serum prostaglandin E(PGE:)and oxygen free radical(MDA)were measured in patients from 2 centers.Results Per-protocol analysis(n=415)showed that the dyspeptic symptom score in rebamipide group decreased significantly after eight weeks of treatment. The endoscopic inflammation score in rebamipide group also decreased from 2.65 ±0.09 to 0.60±0.10(P<0.001),which was,significantly better than that of sucralfate group(P<0.001).Histological findings were consistent with the endoscopic findings.There Was a significant elevation(P=0.002)in PGE2 concentration in mucesa from rebamipide-treated subjects [(225.4±18.3) pg/g vs.(266.7±14.7)Pg/g] compared with that in sucralfate group.The concentration of MDA significantly decreased from(325.9±65.6)mmoL/g to(216.5±61.5)mmol/g,which is markedly different from that of sucralfate group(P=0.046).No statistical difference was found between Hp eradication group,Hp infection group and Hp negative group,regarding the effect of Rebamipide.Conclusion Compared to sucralfate,Rebamipide demonstrates a superior effect on improvement of dyspepsia symptom and endoscopic findings in erosive NAG,which is not influenced by Hp infection.
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Objective To develop a novel endoscopic classification system to determine the invasion depth of early esophageal cancer.Methods The esophageal lesion was endoscopicaUy stained with Lugol's iodine first,then methylene blue.According to the growth pattern,height and cup depth under endoscope,the lesions were classified into 5 types,including surface diffusion growth,intra-lumen growth,intra-wall growth,bi-direction growth and mix growth types.The lesions were then removed by endoscopic mucosa reection or surgery,the precise invasion depth of the lesion was determined pathologically and the results were compared with the endoscopy classification.Results The data of 44 cases of esophageal mucosal cancer and 34 cases of esophageal sub-mueosal cancer were included.With the criteria of mucosal cancer as surface diffusion growth,intra-lumen growth <5mm,bi-direetion growth <2mm and intra-wall growth <0.5 mm,the diagnostic specificity was 89.1%(41/46) and sensitivity was 93.2%(41/44).With the criteria of submucosal cancer as intra-lumen growth≥5 mm,hi-direction growth≥2mm,intra-wall growth≥0.5 mm and mix growth type,the diagnostic specificity was 90.6%(29/32) and sensitivity was 85.3%(29/34).The overall diagnostic accuracy in differentiating esophageal mucosal cancer from esophageal submueosal cancer by endoscopic classification was 89.7%(70/78).Conclusion This endoscopic classification system is effective in differentiating esophageal mucosal cancer from submucosal ones.
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Objective To investigate the endoscopic and ultrasonic endoscopic features of colorectal carcinoid and the indications of endoscopic treatment. Methods The clinical data of 22 patients with colorectal carcinoid who had been admitted to our hospital from 2002 to 2007 were collected. The endoscopic and ultrasonic endoscopic features and the relationship between the features and invasion depth of colorectal carcinoid were analyzed. Results Under the endoscope, early carcinoid presented submucosa tumor with 1.5cm in diameter, and yellow or white smooth surface; advanced carcinoid presented submucosa tumor with 0.8-3.0cm in diameter, and yellow or white little nodus or ulcerative surface. The ultrasonic endoscopic feature of the colorectal carcinoid was orbicular-ovate low level echo tumor with punctiform slightly high-level echo and an unsharpness edge. Sixteen mucosal layer-cancers and submucosal layer-cancers were removed by endoscopic mucosal resection, and 10 of them were additional treated by argon plasma coagulation. After a follow-up period of 4-36 months, no recurrence was observed. Conclusions Endoscopy and endoscopic ultrasonography are effective methods to diagnose colorectal carcinoid and its invasion depth. Endoscopic treatment is a simple, safe and effective means to treat the early colorectal carcinoid tumors.
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Objective To investigate the effect of silence of human protection of telomeres 1 (hPOT1), which was induced by RNA interference, on expression of telomeric repeat factor 1 (TRF1), telomeric repeat factor 2 (TRF2) and Tankyrase 1 in human gastric cancer cell BGC823. Methods The ex-pression of TRF1 ,TRF2 and Tankyrasel at mRNA level were determined by semi-quantitative RT-PCR. Re-sults Significant increase in expression of TRFI, marked decrease of TRF2 and Tankyrase1 at mRNA level were observed in cells of hPOT1 siRNA. Conclusion The significant increase in expression of TRF1 and the marked decease in TRF2 and Tnakyrasel at mRNA level after the inhibited expression of hPOT1 in human gastric cancer cell BGC823 indicate that hPOTI is highly correlated with the expressions of other three te-lomere-specific binding proteins.
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There is a kind of intrinsic correlation between achievements and age.The average years of age of the scientists awarded the Nobel prizes in five fields are 30.9~33.3.Lots of ancient and modern scientists in many fields are similar to this.Therefore young people are able to make significant achievements,especially innovative achievements.This law is not well cogitated in our country,especially by the young themselves.To understand this law correctly will be of certain inspiration and significance not only for people from different age bracket to make achievement but also beneficial to improving the innovative capability of the whole country.
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BACKGROUND: Microsatellite instability (MSI), an important gene change type, plays animportant role in the occurrence of tumor. Mismatch repair gene induces its occurrence. Although the effect of mismatch repair gene hMLH1 mutation in the hereditary nonpolyposis colorectal cancers (HNPCC) has been reported, its effect on the sporadic colorectal carcinoma lacks in-depth study.OBJECTIVE: To investigate the effect of mismatch repair gene hMLH1 mutation on colorectal carcinogenesis, and its correlation with MSI.DESIGN: Single-sample experiment.SETTING: Department of Gastroenterology, Southwest Hospital of Third Military Medical University of Chinese PLA.PARTICIPANTS: Seventy-six cases of sporadic colorectal carcinoma and corresponding normal tissues were obtained from surgically resected specimens of coloreetal carcinoma in Southwest Hospital between January 2001and December 2003. No patients had family history of tumor, or had received radiotherapy and chemotherapy. Patients were informed of the experiment.METHODS: Mutation of hMLH1 was detected by two-dimensional electrophoresis and DNA sequencing; MSI was analyzed by PCR-based methods.MAIN OUTCOME MEASURES: ① Detection rate of hMLH1 mutation of colorectal carcinoma and MSI. ② The relationship of MSI and hMLH1 mutation.RESULTS: Seventy-six cases of sporadic colorectal carcinoma were studied for hMLH1 mutation and MSI. hMLH1 mutation was detected in 8 (10.5%) cases of colorectal carcinomas while MSI was detected in 20 (26.3%) cases of colorectal carcinomas. Frequency of hMLH1 mutation and MSI was significantly higher in right colorectal cancer than in left colorec tal cancer (6/26 vs 2/50, x2=4.739, P=0.029; 11/26 vs 9/50,x2=5.212,P=0.022). No association was observed between hMLH1 mutation or MSI and tumor size, differentiation, histological type, depth of invasion, metastasis or clinical pathological stages. ② MSI was divided into high-frequency group (≥ 2 loci, n=10) and low-frequency group (1 locus, n-10), and MSI negative group (n=56). 8 hMLH1 mutations were all detected in high frequency MSI group, but no mutation was found in low frequency MSI or MSI negative groups.CONCLUSION: hMLH1 mutation and MSI occur in cancer of the right large intestine and hMLH1 mutation is involved in carcinogenesis of some sporadic colorectal cancer with high-frequency 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 investigate the effects of expressions of NF ?B and survivin on the anti tumor activity of TRAIL in gastric carcinoma cells. Methods Gastric carcinoma cells were cultured in RPMI 1640. The apoptotic rates of gastric carcinoma cells SGC 7901, MKN28, AGS, and MKN45 after treatment with TRAIL were analyzed by flow cytometry. The expressions of NF ?B and survivin in the 4 gastric carcinoma cells were determined by Western blotting. Results At 24 h after gastric carcinoma cells exposed to TRAIL at the dose of 300 ng/mL, the apoptosis percent ages were 36 05% for MKN28, 20 27% for MKN45, 16 50% for AGS, and 11 80% for SGC 7901. Western blotting showed that expressions of NF ?B and survivin were lower in MKN28 than those in MKN45, AGS and SGC 7901. Conclusion The anti tumor sensitivity of TRAIL to gastric carcinoma cells is related to the expressions of NF ?B and survivin.
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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 compare the real time diagnosis and treatment values of magnifying endoscopy and electronic colonoscopy Methods A total of 105 colorectal polyps for colonoscopic examination were included in this study A magnifying videoscope with zoom ranges from ?1 to ?100 magnification and a common electronic endoscopy combined with indigocarmine dye were employed to observe the pit patterns of colorectal polyps Pit patterns were analyzed according to Kudo's modified classification as follows: ①type Ⅰ: round pit; ②type Ⅱ: asteroid pit; ③type Ⅲs: tubular or round pit, which is smaller than a normal pit (type Ⅰ); ④type ⅢL: tubular or round pit, which is larger than a normal pit (type Ⅰ); ⑤type Ⅳ: dendritic or gyrus like pit; ⑥type Ⅴ: irregular or amorphous pit; and ⑦ mixed type Results Magnifying colonoscopy revealed that phenotypes of non neoplastic and neoplastic lesions were 78 57% and 21 43% in inflammatory and hyperplastic polyps, 3 33% and 96 67% in neoplastic polyps, and 100% non neoplastic phenotype in juvenile polyps, respectively Pit pattern analysis according to Kudo's modified classification showed that the diagnostic sensitivity of neoplastic and non neoplastic lesions was 96 67% and 80%, and specificity was 86 57% and 94 73%, respectively The overall diagnostic accuracy in differentiating neoplastic from non neoplastic lesions was 89 52% Pit pattern by common electronic colonoscopy showed that the diagnostic sensitivity of neoplastic and non neoplastic lesions was 88 3% and 73 3%, and specificity was 81 5% and 82 5%, respectively The overall diagnostic accuracy in differentiating neoplastic from non neoplastic lesions was 82% Conclusion The pit pattern analysis of colorectal lesions by magnifying colonoscopy or electronic endoscopy combined with indigocarmine dye is a useful method for the identification of non neoplastic polyps, adenomas and invasive carcinomas in the large bowel Therefore, it may be possible to determine, at the time of colonoscopy, which lesions should be removed endoscopically and surgically
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Objective To investigate the effect of membrane-cytoskelet on linker Ezrin in the invasion and migration of human adenocarcinoma cells line AGS. Methods Two small hair RNA targeting Villin2 gene ( Ezrin coding gene) was designed,chemically synthesized and inserted into plasmid pGenesil-shRNA,then transformed into E. coli DH5 ?. The recombinant plasmid was extracted in middle quantity and transfected into AGS cells by LipofectamineTM 2000. Ezrin expression in AGS cells transfected by shRNA recombinant plasmid was measured by RT-PCR and Western blotting. AGS cells were transfected by effective shRNA plasmid and cultured in 1640 media containing G418 ( 800 ?g/ml) . RT-PCR and Western blotting were applied respectively to detect the Ezrin mRNA and protein expression to evaluate the blocking effect of shRNA-Ezrin. Cell migration and invasion was determined by scratch test and Transwell chamber assay. Results Our designed shRNA knocked down 93% Ezrin in AGS cells which had a stable expression of Ezrin small hairpin RNAs. The migrated cells number of Ezrin knocked-down group was 27. 67 ? 4. 50,which was significantly decreased compared with the control group ( 125. 50 ? 8. 33,P
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Objective To investigate the expression and significance of Claudin 3 in adenocarcinoma of esophagus and gastric cardia.Methods We subjected pathologically confirmed 16 cases of esophageal adenocarcinoma(EAC),20 cases of esophageal squamous cell carcinoma(ESC),and 40 cases of gastric cardiac adenocarcinoma(CAC,including 15 diffuse and 25 intestinal cases).Tissue samples were collected with aid of gastroscope.Fifteen samples of normal esophageal mucosa and 15 samples of normal cardiac mucosa served as control.RT-PCR and Western blotting were used respectively to detect the mRNA and protein expression of Claudin 3 in these tissue samples.Results The mRNA and protein expressions of Claudin 3 were increased in EAC and intestinal CA compared with those in normal tissues,ESC and diffuse CAC.There was no significant difference of the expression of Claudin 3 between normal group and ESC and diffuse CAC group.Conclusion Claudin 3 might be related to the development of EAC and intestinal CAC,and expected to be the tumor maker and therapeutic target of this kind of carcinoma.
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Objective To investigate the polymorphism of peroxisome proliferator-activated receptors-?2(PPAR-?2) and its association with obesity in Chinese population.Methods According to BMI,89 subjects who are in normal body weight range and 116 overweighted and obese subjects were included in this study.Their Pro12Ala mutation in the PPAR-?2 gene was detected by PCR restriction fragment length polymorphism.Results Allele frequency of Ala mutation of PPAR-?2 in overweighted and obese subjects(qA=11.64%) was significantly higher than that of normal body weight group(qA=5.06%,P