Objective To introduce the methods and the advancements of early diagnosis in primary carcinoma of gallbladder (PCG), and improve the early diagnostic rate of PCG. Methods Recent relevant literatures were reviewed. Results It was difficult in early diagnosis of PCG and with a poor prognosis. Comprehending case history and careful examination and being assisted by multiple imaging methods and molecular biology technology could markedly improve the early diagnostic rate. Conclusion Comprehending the progress will contribute a lot of improving the early diagnostic rate and selecting reasonable clinical methods to be used in early diagnosis of PCG.
【Abstract】ObjectiveTo analyze the factors influencing the prognosis of patients with bile duct carcinoma after resection. MethodsThe clinical data of 120 patients with bile duct carcinoma receiving resection in our hospital from 1980 to 2004 were collected retrospectively and clinicopathologic factors that might influence survival were analysed. A multiple factor analysis was performed through Cox proportional hazard model. ResultsThe overall 1year, 3year and 5year survival rates were 71.7%, 32.5% and 19.2% respectively. The single factor analysis showed that the major significant factors influencing survival of these patients were histological type of the lesions, lymph node metastasis, pancreatic infiltration, duodenal infiltration, resected surgical margin, perineural infiltration, peripheral vascular infiltration and depth of tumor infiltration (P＜0.05). Lymph node metastasis, pancreatic infiltration and perineural infiltration were found to be the the statistically significant factors influencing survival by multiple factor analysis through the Cox model. ConclusionThe most important prognostic factors for bile duct carcinoma after resection were lymph node metastasis, pancreatic infiltration and perineural infiltration.
Objective To obtain the polypeptides specifically binding to gastric cancer BGC823 cell from a Ph.D-12 TM phage display peptide library (PDPL), to search the markers of gastric cancer for early diagnosis and treatment. Methods The gastric cancer BGC823 cell was used as the antigen and the immortalized gastric epithelial GES cell was used as control for 3 rounds subtraction biopanning from PDPL at room temperature. The positive and specific binding clones were identified by cell enzyme-linked immunosorbent assay (ELISA) and immunochemistry staining. Those DNA sequences of identified clones were sequenced, polypeptides were marked by fluorescein FITC peptide tag, and polypeptides affinity and specificity of gastric cancer cells and tissues were identified. Results After 3 rounds of panning, 20 phage clones were identified by ELISA, one of which （GC-11） was specially binding to the BGC823. Cell and tissue immunofluorescence assay further presented a high affinity of fluorescein-labeled peptide FITC-GC-11 with BGC823 and gastric cancer tissue.Conclusion A peptide GC-11 which is specific binding to gastric cancer BGC823 cell and gastric cancer tissue has been selected from PDPL.
Objective To observe the expression of Ezrin protein in primary carcinoma of gallbladder tissue and the levels of CEA and CA19-9 in serum of patients with primary carcinoma of gallbladder, and to explore the relationship between the expressions of these measurements and clinicopathologic characteristics. Methods Immunohistochemistry was applied to analyze the expression of Ezrin protein in primary carcinoma of gallbladder and chronic cholecystitis tissue. The levels of CEA and CA19-9 in serum and clinicopathologic characteristics of all including patients were detected with clinical measurement. All data were analyzed statistically. Results ①The positive rates of Ezrin protein in primary carcinoma of gallbladder and chronic cholecystitis tissue were 66.7% （40/60） and 30.8%（4/13）, respectively (χ2=5.57, Plt;0.05). ②There was no difference between the expression of Ezrin protein in primary carcinoma of gallbladder tissue and age or gender (Pgt;0.05). However, difference was significant between the Ezrin expression and degree of difference, pNevin stages, pTNM stages, lymph node metastasis or distant metastasis (Plt;0.05). ③There were no differences between the positive rates of CEA and CA19-9 in primary carcinoma of gallbladder and age or gender (Pgt;0.05). However, differences were significant between the positive rates of CEA and CA19-9 and pNevin stages, pTNM stages, degree of difference, lymph node metastasis or distant metastasis (Plt;0.05). ④There was some relationship between the expression of Ezrin protein and the positive rate of CEA (rs=0.213, Plt;0.05), but not with the positive rate of CA19-9 (rs=0.081, Pgt;0.05). Conclusions The high expression of Ezrin protein may promote the invasion and metastasis in primary carcinoma of gallbladder. It could be possible to decide the outcome of primary carcinoma of gallbladder through the combined analysis on the expression of Ezrin protein and the serum levels of CEA and CA19-9.
Objective To study the effects of L-arginine （L-Arg） on cell proliferation, inducible nitric oxide synthase (iNOS) expression and cell cycle in human colon carcinoma cell line LS174 through nitric oxide (NO) pathway. Methods LS174 cells were cultured in medium with L-Arg at different concentrations for different times. MTT method was employed to evaluate the level of the cell proliferation. The production of NO in culture supernatants of LS174 cell was detected with enzyme reduction of nitrate. The distribution of the cell cycle was detected with the flow cytometry (FCM). The expression level of iNOS in the cells was determined by Western blot and SP immunocytochemical staining method. Results The growth of LS174 was promoted by the L-Arg at low concentration (0.125 mmol/L) and inhibited at high concentrations (0.5， 2， 8 and 32 mmol/L). The level of NO was increased with the increasing concentration of L-Arg in culture medium. To compare with the control group, the ratio of cells at S phase was increased after 48 hours’ treatments with high concentrations (0.5, 2, 8 and 32 mmol/L) of L-Arg (P＜0.05, P＜0.01); while there was no obvious difference after treatments with low concentration (0.125 mmol/L) of L-Arg (Pgt;0.05). With the increase of the concentration of L-Arg, the expression of iNOS was increased as compared with control group. The higher the concentration of L-Arg was, the better the effect. Conclusion L-Arg can induce the expression of iNOS resulting in increase the production of nitric oxide (NO). Low concentration of L-Arg can promote the growth of LS174 cells, while high concentration ones can inhibit growth and proliferation. The high concentration of L-Arg could induce S phase arrestion in the cell cycle.
Objective To design and construct the eukaryotic expressed vector of suicide genes, which contained 5 copies of hypoxia-responsive element (5HRE), promoter of alpha-fetoprotein gene (AFPp） and nitroreductase from Escherichia coli. Methods The constructing processes were as follows: ①The design of primer: Suicide genes of NTR in the Escherichia coli, which contained 6his-tag gene (6his-tag), were cloned by overlapping PCR. ②The construction of 5HRE: The single strand of synthetized HRE oligonucleotide was annealed, and 5HRE was constructed by multiple recombinant clone. ③The recombination of NTR gene, 5HRE, AFPp and pIRES2-EGFP: pIRES2-EGFP, which had removed the instant early promoter of cytomegalovirus, was recombined with NTR gene, 5HRE, AFPp. In this way, the eukaryotic expressed vector of pIRES2-EGFP-5HRE-AFPp-NTR, which carried NTR gene, 5HRE, AFPp was finally constructed. Results NTR gene, which contained the fusion of 684-base pair and 6his-tag gene, was cloned successfully, and its sequence was coincident with the result published by Genbank. A 221-base pair of 5HRE was also constructed, which was in accordance with the expected sequences. The integrity of the eukaryotic expressed vector was verified by restriction enzyme digestion and DNA sequence analysis, respectively. Conclusion The eukaryotic expressed vector of pIRES2-EGFP-5HRE-AFPp-NTR is successfully constructed, which may be used for its further investigation in vitro.
【Abstract】ObjectiveTo evaluate the advances in minimally invasive surgery treatment for hepatolithiasis. MethodsLiteratures about the advances in minimally invasive surgery treatment for hepatolithiasis were collected and reviewed. ResultsHepatectomy and bile duct exploration using laparoscopy can get better effect. Fibrocholedochoscopy play an important role in the course of operation and after operation for hepatolithiasis. ConclusionThe individualization treatment program should be used for hepatolithiasis. Association application of multipathway minimally invasive operation, such as laparoscopy, fibrocholedochoscopy and so on, can increase the cure rate of hepatolithiasis.
【Abstract】ObjectiveTo study the relationship between the bacterial spectrum difference of gallbladder mucosa and choledochus bile and clinical prognosis of gallstone pancreatitis. MethodsA synchronic bacterial culture and drug sensitivity test were carried out on 48 patients with gallstone pancreatitis. ResultsThe cases of positive gallbladder mucosa and choledochus bile were 13 (27.1%) and 31 (64.6%) respectively. The cases of double positivity were 12 (25.0%). The cultural strains of gallbladder mucosa and choledochus bile were significantly different. Some strains were only found in choledochus bile，whereas in gallbladder mucosa L-form bacteria predominated.ConclusionThe most common causative strain of gallstone pancreatitis is Bacterium coli. The drug-resistant strain emerges maybe due to bacterium immigration and delitescence in gallbladder mucosa. L-form bacteria should be considered when using antibiotics, because L-form bacteria have close relationship with the prognosis of gallstone pancreatitis.
ObjectiveTo study the mechanism of the effect on invasion and metastasis of colorectal cancer by down-regulating c-Met gene.Methodsc-Met genes were knocked down in SW480 cells, differential genes were screened by gene chip, functional cluster analysis of differential genes was carried out, and IPA was used to analyze the interaction network of cell signal pathway and related differential genes, as well as the ralationship between related genes and upstream regulatory molecules. The related genes in the suppressed signal pathway were selected for qPCR verification.ResultsAfter knockdown of c-Met, the number of up-regulated genes and down-regulated genes in SW480 cells was 399 and 286, respectively. Cluster analysis showed that c-Met knockdown had a great effect on the gene expression level of SW480 cells, IPA pathway analysis showed that HGF signaling pathway was suppressed, and after c-Met knockdown, IPA interaction network suggested that AKT2, PIK3CA, and MAP2K4 in HGF pathway were down-regulated, and qPCR verified that the above genes were also down-regulated, which was consistent with the results of microarray.Conclusionc-Met may affect the invasion and metastasis of colorectal cancer through the regulation of AKT2, PIK3CA, and MAP2K4 in HGF pathway.
ObjectiveTo investigate the correlations among the cadual homeobox gene 2 (CDX2), hypoxia inducible factor-1α (HIF-1α) protein expressions, and tumor budding in the colorectal cancer (CRC). MethodsIn this study, 63 CRC specimens surgically removed in the First Affiliated Hospital of Xi’an Jiaotong University from January 2012 to September 2015 were collected. The CDX2 and HIF-1α protein expressions were detected by immunohistochemical staining streptavidin-biotin peroxidase two-step method. The staining and the grade of tumor budding were observed under an optical microscope, and the correlation was analyzed using Spearman test. ResultsThe positive expressions of CDX2 and HIF-1α proteins in the CRC tissues were 35 (55.6%) and 47 (74.6%) cases, respectively, which was a negative correlation in the CRC (rs=–0.302, P=0.017). The positive expressions of CDX2 and HIF-1α proteins in the tumor budding of colorectal cancer were 21 (51.2%) and 26 (63.4%) cases, respectively, which was also a negative correlation in the tumor budding of CRC (rs=–0.336, P=0.031), but there was no statistic correlation between the grade of tumor budding and CDX2 or HIF-1α positive protein expression in the CRC (rs=0.113, P=0.370; rs=–0.026, P=0.838). ConclusionsThe positive expression between CDX2 and HIF-1α has a negative correlation in the same CRC specimen and which has a negative correlation in tumor budding. There is no statistic correlation between grade of tumor budding and CDX2 or HIF-1α protein expression in the CRC. Hypoxia environment may be involved in the downregulation of CDX2 level during the malignant progression of CRC.