Objective To explore the pathogenesis of the level of gene and therapeutic target genes associated with intestinal obstruction by analyzing the differential expression gene. Methods The gene expression data that came from public database gene expression omnibus （GEO） which provided adhesion formation’ gene expression data on 1， 3， 7，and 14 days after operation （n=8） and normal intestinal tissues’ gene expression data （n=2） of mouse were collected. The gene function and differential expression of genes were analyzed by using gene ontology （GO） and significance analysis of microarray （SAM）. Results There were a lot of response stimulated up-regulation of gene expression when occurrence of adhesion， and the products of these genes were distributed on cell membrane. The analysis results of gene expression at different time point after operation showed that expression up-regulated of Hmgcs 2 gene occurred on 3-14 days ofter operation and expression up-regulated of Stxbp 5 gene occurred on 14 days ofter operation. Conclusions The adhesion formation may be closely associated with the genes of response to stimulus and the gene product in membrane. The Hmgcs 2 and Stxbp 5 genes may be closely associated with the occurrence of other diseases which induced by adhesion formation.This provides a basis for the discovery of potential therapeutic targets.
Objective To summarize the current advancement of preoperative radiotherapy for rectal cancer. Methods Relevant literatures about current advancement of preoperative radiotherapy for rectal cancer published domesticly and abroad recently were collected and reviewed. Results The lower local recurrence rate and longer disease-free survival time were observed in preoperative radiotherapy， compared with postoperative radiotherapy for rectal cancer. The recurrence rate was higher in short-course radiotherapy， compared with conventionally radiotherapy for stageⅢrectal cancer， but there was no significant difference for stageⅡrectal cancer. The biology molecular such as p53， CEA， Cox-2， EGFR， and VEGF had shown to be radiosensitive. Conclusions The proposal of preoperative radiotherapy for rectal cancer， could be prone to conventionally radiotherapy. There are more screening targets for preoperative radiotherapy in extensive exploration of diverse radiosensitivity. Biology molecular， developed gene expression profiling， and gene chips for rectal cancer may contribute to the individualization treatment.
【Abstract】ObjectiveTo detect p27 expression in rectal carcinoma and serum transforming growth factor-β1 (TGF-β1) level in these patients, and to elucidate the modulatory effect of serum TGF-β1 on p27 expression in rectal carcinoma. MethodsExpression of p27 was measured in 37 cases of rectal carcinoma, 22 of rectal adenoma and 19 of normal control specimens by immunohistochemical staining using antibodies to p27. Serum level of TGFβ1 was measured in these patients by enzymelinked immunosorbent assay (ELISA) method. Resultsp27 protein was expressed in normal rectal tissue, rectal adenoma and rectal carcinoma, and the positive rate was 89.47%, 90.91% and 64.87%, respectively. The positive rate of p27 in rectal carcinoma was significantly lower than that of normal rectal tissue and rectal adenoma (P=0.025). p27 was mainly located in nucleolus of normal rectal tissue and rectal adenoma, and the positive rate of p27 in cytoplasm of rectal carcinoma was higher than that of normal and rectal adenoma. The positives rates of serum TGF-β1 in normal group, rectal adenoma group and rectal carcinoma group were 21.05%, 27.27% and 51.35%(P=0.045),respectively. The expression of p27 related to histological differentiation, lymph node metastasis and infiltration depth. Serum level of TGF-β1 related to lymph node metastasis, infiltrated depth and CEA level. The positive rate of p27 in TGF-β1 negative group and positive group was 88.89% and 42.11%(MantelHaenszel χ2=6.755,P=0.009), respectively. ConclusionTGF-β1 may be useful in assessment of malignance and prognosis of rectal carcinoma. TGF-β1 can downregulate p27 expression in rectal carcinoma.
【Abstract】ObjectiveTo study the effect of transfection with antisense DNMT3b gene eukaryotic expression vector on the expression of DNMT3b gene in human cholangiocarcinoma cell line QBC-939. MethodsThe constructed antisense DNMT3b gene eukaryotic expression vector was transfected into the human cholangiocarcinoma cell line QBC-939 by using lipofectamine transfection reagents, and positive cell clones were obtained by using G418 selection after transfection. Whether the constructed recombinant vector was transfected into QBC-939 cells successfully was confirmed by amplifying the exogenous neoR gene with PCR method. The expression of DNMT3b gene mRNA and protein were detected by semi-quantitative RT-PCR and FCM methods respectively. ResultsFollowing the transfection of antisense DNMT3b gene eukaryotic expression vector, the mRNA level of DNMT3b gene in QBC-939 cells of human cholangiocarcinoma decreased from 0.956±0.053 to 0.209±0.023, and the protein level of DNMT3b gene also decreased from (75.38±3.22)% to (29.87±3.46)%. There were very significant differences on the expression levels of DNMT3b gene between non-tranfections group and the antisense DNMT3b gene eukaryotic expression vector transfection group (P＜0.01). ConclusionTransfection with antisense DNMT3b gene eukaryotic expression vector significantly reduces the expression level of DNMT3b gene in human cholangiocarcinoma cell line QBC-939, and this study may provide a valid tool and method to investigate the function of DNMT3b gene and its role in cholangiocarcinoma.
【Abstract】Objective To establish and assess the rat model of postoperative fatigue syndrome (POFS). Methods The rat model of POFS was developed by the partial resection of the liver. The behavioral changes prior and post to operation, the disorder of nutritive intake after operation, stress reaction (pathological changes of mucous membrane in small intestine) and the hepatic albumin gene expression were observed. Results Low body temperature, lower sensitivity and reactivity were found. The serum levels of the iron, total protein, albumin, globulin and so on as the indexes of nutrition obviously dropped. The injury of the mucous membrane resulted from the stress reaction after the resection of the liver. The gene expression of the albumin decreased in the model group.Conclusion The experimental rat model of POFS by partial resection of the liver can be used for the investigation of POFS.
【Abstract】Objective To design the hammerhead ribozyme gene according to the hTR sequence in the gallbladder cancer cell, and build it into the eukaryon expression vector pTriEx-4. Methods According to the hTR cDNA sequence, the authors designed the primers and take the hTR template area gene from the gallbladder cancer cells by RT-PCR.The hammerhead ribozyme gene was synthesize according to the result of sequencing, and combine them with eukaryon expressing vector. Identified the exactitude of recombine vector by digestion.Results The 68 bp sequence extracted from the cell through the RT-PCR had the same template sequence comparing with the hTR cDNA. The recombinant plasmid with the hammerhead ribozyme gene was correct by digestion identification. Conclusion The RT-PCR method can extract the gallbladder cancer cell’s hTR gene. We construct the eukaryon expression vector containing the hammerhead ribozyme gene successfully which is the foundation for gene therapy of gallbladder cancer.
ObjectiveTo study the relationship between expression of p27KIP1 and progression of hepatocellular carcinoma(HCC).MethodsThe expression of p27KIP1 in 52 cases of HCC was detected by immunohistochemistry of strept avidinbiotin complex and mRNA in situ hybridization.ResultsThe positive cells of p27KIP1 protein were diffused in HCC.The positive signal was localized in nuclei.The labeling index (LI) of p27KIP1 protein was significantly higher in tumorsurrounding tissues than that in tumor tissues. p27KIP1 protein LI showed a positive correlation with the differentiation grade of HCC.The better differentiation of cancer cells, the higher LI of p27KIP1 protein (P＜0.01).The positive cells of p27KIP1 mRNA were also diffused in HCC.The positive signal was localized in nuclei and cytoplasm. As to the expression of p27KIP1 at the mRNA level,there was no significant correlation with tumorsurrounding tissues and stages of HCC.Conclusionp27KIP1 protein is associated with progression and differentiation grade of hepatocellular carcinoma.
Objective To investigate the effect of mRNA expression of gelatinase A on the invasion and metastasis of human gastric carcinoma (HGC). MethodsThirtysix cases of HGC were examined by in situ hybridization technique. ResultsPositive expression rates of gelatinase A in the normal gastric tissue, peritumor tissue and HGC were 8.3%,35.7% and 83.3% respectively （P＜0.01）. The positive rates of gelatinase A in the group with serosal invasion and lymph node metastasis were 93.1% and 90.6%, much higher than those in the group with negative ones (42.9% and 25.0%).By in situ hybridization, gelatinase A mRNA was showed to be expressed in the extracellular matrix of tumor tissues,which surrounded the invasive margin of cancer tissues. The positive cells at these sites were mainly tumorinfiltrating macrophages. Conclusion There is good correlation between gelatinase A mRNA expression and the invasion, metastasis of HGC. So it can be used as a useful marker for invasion and metastasis of HGC.
Objective To investigate the regulatory effect of somatostatin analogue (SMS201995,SMS) on proliferation and apoptosis in human cholangiocarcinoma cell line in vitro. MethodsProliferation curve, flow cytometry, agarose gel electrophoresis, Annexin VFITC and flow cytometric immunofluorescent technique were performed to identify the inhibitory effect on cell proliferation and the induction of apoptosis of human cholangiocarcinoma cells (SKChA1). ResultsSMS significantly reduced the SKChA1 cell growth by serum in long experiments and transiently accumulated it in G0/G1 phase. Dotplot analysis of cells duallabeled with Annexin VFITC and PI confirmed the induction of apoptosis by SMS in SKChA1 cells.AnnexinVFITC labeling was markedly enhanced following treatment with SMS for 24 h. DNA of treated SKChA1 cells appeared a ladder pattern characteristic of apoptosis. Besides, timedependent increase in bax and decrease in bcl2 occured during SMS treatment. Conclusion SMS could inhibit the proliferation activity and induce apoptosis of cholangiocarcinoma cell line SKChA1. The mechanisms of apoptosis might be correlated with the expression of apoptosisregulatory gene bax and bcl2.
We have used immunphistochemical SP method to detect the expression of nm23 gene protein in human thyroid tissues from 86 carcinomas, 20 adenomas and 25 carcinomatous adjecent tissues. The results showed the positive staining rate were 73.3%, 40.0%and 16. 0% respective (Plt; 0. 005). Although the expression of nm23 protein had no association with the cervical lymph node metas-tases,it was significantly concordant with the tumor cell diffierentiation (Plt;0. 01) ,tumer capsule (Plt; 0. 05) and TNM stage (Plt;0.05).In addition, the patient‘s average survival time in nm23-positive cases was longer than that in nm23-negative ones (Plt;0.01).This data suggest that nm23 gene may play an important role in thyroid carcinogenesis and the expression of nm23 protein would be an useful marker in assessing the prognosis of the thyroid carcinomas.