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Colorectal carcinogenesis is the result of accumulation of genetic and epigenetic alters.Gene mutations and changes in DNA methylation patterns,resulted from genomic instability,are the main molecular events in colorectal cancer.Identification of key mutations and methylation phenotypes in genes leading to colorectal cancer progression,and molecular pathology classification and diagnosis are likely to be the prerequisites for individualized and targeted treatment.
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Objective To over-express human trefoil factor 2 (hTFF2) by Escherichia coli system and an-alyze its activities in promoting migration and anchorage-independent growth in SW480 colonic cancer cells. Meth-ods hTFF2 gene encoding mature peptide was obtained by RT-PCR, and the recombinant expression vector pET32a-hTFF2 was constructed. Then pET32a-hTFF2 was transformed into E. coli BL21-32a and TrxA-hTFF2 fu-sion protein was induced to over-express. The expressed product was isolated by Ni-NTA affinity chromatography, purified by dialysis and identified by Western blotting. The activities of the recombinant hTFF2 in promoting SW480 cells migration and anchorage-independent growth were analyzed by MicroChemotaxis Chamber migration assay and Soft-agar assay,respectively. Results The TrxA-hTFF2 fusion protein was expressed to 220 mg/L at high purity. In vitro model demonstrated that recombinant hTFF2 obviously enhanced SW480 cell migration activity and anchor-age-independent growth. Conclusion The recombinant hTFF2 can be expressed in E. coli with high production, purity and biological activities. And its roles in cell migration and anchorage-independent growth suggest that up-regulation of TFF2 in colonic cancer might be involved in cancer invasion and metastases.
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Objective To investigate the related risk factors of prognosis in patients with systemic lupus erythematosus (SLE) complicated with lung infection.Methods The clinical data of 84 cases with SLE complicated with lung infection were analyzed retrospectively.Multivariate Logistic regressive analysis was made with death and survival as the independent variable.Results There were 23 out of 84 patients dead,and fatality rate was 27.38％.Multivariate Logistic regressive analysis showed that the urine volume,immune globulin treatment,cyclophosphamide pulse therapy,plasma albumin and SLE disease activity (SLEDAI) score were the risk factors that influenced the prognosis (OR =0.53,0.72,4.29,0.94,1.58,respectively,P＜0.0l or ＜0.05).Conclusions The prognosis of patients with SLE complicated with lung infection is the result of the joint action of many factors.The higher SLEDAI score and cyclophosphamide pulse therapy are the independent risk factors of the prognosis.The increase of plasma albumin level and urine production and immune globulin treatment are helpful to decrease the fatality rate.
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Intestinal barrier dysfunction plays an important role in spontaneous bacterial peritonitis. In the present study, changes in the intestinal barrier with regard to levels of secretory immunoglobulin A (SIgA) and its components were studied in fulminant hepatic failure (FHF). Immunohistochemistry and double immunofluorescent staining were used to detect intestinal IgA, the secretory component (SC) and SIgA in patients with FHF (20 patients) and in an animal model with FHF (120 mice). Real-time PCR was used to detect intestinal SC mRNA in the animal model with FHF. Intestinal SIgA, IgA, and SC staining in patients with FHF was significantly weaker than in the normal control group (30 patients). Intestinal IgA and SC staining was significantly weaker in the animal model with FHF than in the control groups (normal saline: 30 mice; lipopolysaccharide: 50 mice; D-galactosamine: 50 mice; FHF: 120 mice). SC mRNA of the animal model with FHF at 2, 6, and 9 h after injection was 0.4 ± 0.02, 0.3 ± 0.01, 0.09 ± 0.01, respectively. SC mRNA of the animal model with FHF was significantly decreased compared to the normal saline group (1.0 ± 0.02) and lipopolysaccharide group (0.89 ± 0.01). The decrease in intestinal SIgA and SC induced failure of the intestinal immunologic barrier and the attenuation of gut immunity in the presence of FHF.
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Animals , Humans , Mice , Immunoglobulin A, Secretory/immunology , Liver Failure, Acute/immunology , Case-Control Studies , Fluorescent Antibody Technique , Immunohistochemistry , Immunity, Mucosal/immunology , Immunoglobulin A, Secretory/metabolism , Intestinal Mucosa/immunology , Liver Failure, Acute/metabolism , Mice, Inbred BALB C , Real-Time Polymerase Chain ReactionABSTRACT
Objective To study the clinical significance and the expression of Twist protein in non-small cell lung cancer(NSCLC).Methods The expression of Twist protein was examined in formalin-fixed paraffin-embedded tissue samples of 61 NSCLC by S-P immunohistochemical method.Results The expression level of Twist in NSCLC was significantly higher than that of normal lung tissue;the positive expression of Twist protein was related to the histotype,clinical stage and lymph node metastasis;the expression level of Twist protein in the cases whose survival time was less than 4 years was significantly higher than that in the cases whose survival time more than 4 years;the Cox analysis revealed that smoking and lymph node metastasis were the noticeable affective factors on the death of the NSCLC patients after surgery.Conclusion s Twist gene played an important role in the genesis and development of NSCLC,and was related to the prognosis.
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Background and purpose:Twist has been identifi ed as tumor metastasis promoter transcription factor and KiSS-1 has been identified as tumor metastasis suppressor gene,and both of them have been identified to be associated with the metastatic potential of non-small cell lung cancer(NSCLC) .Our aim was to identify the expression of both Twist and KiSS-1 in NSCLC and analyze their correlation with patients’ survival.Methods:Immunohistochemical staining using monoclonal Twist and KiSS-1 antibody were performed on paraffi n embedded specimens from 61 patients diagnosed with NSCLC,and 15 specimens of tumor surrounding lung tissue were used as control.The association with clinicopathologic data and prognosis of NSCLC were analyzed.Results:The expression of Twist was significantly higher in NSCLC than in tumor surrounding lung tissue(P
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<p><b>OBJECTIVE</b>To detect expression of p-glycoprotein (P-gp), multidrug resistance-associated protein (MRP), and glutathione S-transferase (GST-pi), and to evaluate its clinical significance in neuroblastoma (NB).</p><p><b>METHODS</b>SP immunohistochemical technique was used to investigate expression of P-gp, MRP, and GST-pi in 70 cases of NB.</p><p><b>RESULTS</b>The frequency of expression of P-gp, MRP, and GST-pi was 61.4%, 38.6%, and 51.4%, respectively. The coexpression rate of P-gp and MRP, P-gp and GST-pi, MRP and GST-pi, P-gp, MRP and GST-pi was 32.9%, 35.7%, 27.1%, and 24.3%, respectively. Significant positive correlation was observed between P-gp and MRP expression (P = 0.001), and between MRP and GST-pi expression (P = 0.012), but no correlation was found between P-gp and GST-pi expression. The expression of P-gp and MRP was higher in tumors from patients over 1 year old compared with those less than 1 year old at diagnosis (P = 0.01, 0.018, respectively). MRP expression was higher in tumors from the metastatic than the non metastatic groups (P = 0.015). All tested proteins showed significant relationship to the differentiation of the tumor (P = 0.006, 0.000, 0.019, respectively), but no correlation was found to the stage of NB or sex of the patients. MRP expression was significantly related to the reduction of both median survival time and the two-year cumulative survival (P = 0.02). In contrast, P-gp and GST-pi expression had no correlation with survival.</p><p><b>CONCLUSIONS</b>The intrinsic multidrug resistance of NB involves the combined effects of P-gp, MRP, and GST-pi. MRP expression may be an important parameter in predicting the prognosis of patients with NB.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Gene Expression , Glutathione S-Transferase pi , Glutathione Transferase , Immunohistochemistry , Isoenzymes , Multidrug Resistance-Associated Proteins , Neuroblastoma , Metabolism , Mortality , Survival RateABSTRACT
Objective To study the effect of BCL-2 ?-radiation on BCL-2 gene in dogs, and its relationship and signifcane on apoptosis of proliferated smooth muscle cells of bile duct wall. Methods The ~(103)Pd (radioactivity) stent(experiment group) or ordinary stent(control group) was positioned into the target segment of bile duct. The injured bile duct segments were dissected free from the dogs, and BCL-2 gene in the (control) and r-radiation-induced apoptotic smooth mucle cells of bile duct wall was analysed by using (immuno-histochemical) technique. The number of apoptotic cells was counted, and size of lumen of bile duct in both groups was measured by a computerized imaging system.Results BCL-2 gene expression was weaker in the ~(103)Pd radioactive stent group than in the ordinary stent group. The group of dogs with low expression of BCL-2 genes showed marked apoptosis of proliferated smooth mucle cells of bile duct and there was no overt stenosis of extrahepatic bile ducts. The group that showed high expression of BCL-2 gene did not show marked apoptosisi of proliferated smooth muscle cells of bile duct, and there was marked stenosis of extrahepatic bile duct.Conclusions The expression level of BCL-2 in experimental dogs is related to the develoment of (cellular) apoptosis and to radiation sensitivity of the cells. ~(103)Pd radioactive stent can reduce the expression of BCL-2 gene, promote apoptosis of proliferated smooth muscle cells of bile duct, and suppress stricture (formation) of extrahepatic bile duct.
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Objective To evaluate the diagnostic value of MSCT longitudinal reconstruction in the ureteral obstruction.Methods 34 patients with ureteral obstruction were scaned transversely and longitudinally by GE Lightspeed 16 CT scanner and the images were reconstructed with three-dimensional software.All cases were proved by operation and pathology except for 2 cases of uretal obstruction compressed by retroperitoneal lymph nodes.Results In 34 cases,there were 22 cases of ureteral calculi,2 cases of ureteritis,3 cases of carcinoma of ureter,3 cases of pelvisureteral conjunction stricture,1 case of ureteral cyst,1 case of retrocaval ureter and 2 cases of ureter compressed by retroperitoneal metastatic lymph nodes.On longitudinal reconstruction images,22 lesions of calculi in the ureter and thickened uretal wall were showed.Contrast-enhanced CT scan was preformed in 5 cases,the CT values of lesions were increased about 20~35 HU,after administration of contrast medium,and light hydronephrosis in 18 cases,medium hydronephrosis in 6,serious hydronephrosis in 2 and no hydronephrosis in one case were showed on delayed contrast-enhanced scan.Conclusion Longitudinal reconstruction image is of significant value in diagnosis of ureteral obstruction.
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Objective To investigate the preventive effects of 103 Pd radioactive stent on stenosis after bile duct injury in dogs.Methods Twelve healthy dogs (15～20kg) were randomly divided into 103 Pd radioactive stent group (n=6) and control group (n=6). Immediately after balloon dilatation injury to the bile duct, the 103 Pd radioactive stent(experiment group) or the ordinary stent(control group) was positioned into the target segment. The dogs were killed one month later. The injured bile duct segments were dissected free from the dogs, and were examined radionucleonically, immunohistochemically and pathologically. Muscular proliferation area and lumen area were determined by computer assisted picture analysis system. Results In the control group, 30 days after ductal injury, the mucosa of the bile duct was fractured, the mucosa was proliferated and the lumen stenosed.Compared with the control group, 103 Pd radioactive stent significantly reduced muscular proliferation area (P