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AIM:To investigate the construction of oligonucleotide microarray specialized for pancreatic adenocarcinoma-associated genes and its preliminary application of detecting differential expressed genes in pancreatic cancer.METHODS:Pancreatic cancer related genes were purposely selected,and oligonucleotide microarray was prepared by spotting oligonucleotide probes on glass slides coated with APS-PDC.Labeled cDNA targets for hybridizations were synthesized by reverse transcription from total RNA in the presence of Cy5-dCTP and Cy3-dCTP,respectively.Hybridized microarray was scanned by Agilent laser scanner,and the aquired image was analyzed by Imagene3.0 software.The intensity ratios of Cy3 and Cy5 were calculated.To confirm the expression profiles of these genes,quantitative reverse transcription-PCR (QRT-PCR) was carried out for CDC25B and TUSC3 genes,and β-actin gene was taken as internal control.The product of PCR was quantitated by comparative Ct method.RESULTS:The signal of microarray hybridization was clear,and the images had a lower background and higher signal-noise ratio.In comparison with normal pancreas,twenty -four differentially expressed genes were identified which included seventeen up-regulated and seven down-regulated genes.The results of QRT-PCR demonstrated that the expressions of CDC25B and TUSC3 in pancreatic cancer were up-regulated and down-regulated respectively,which is consistent with microarray results.CONCLUSION:The oligonucleotide microarray specialized for pancreatic cancer is desirable for its specialty and sensitivity,which can simultaneously and parallelly detect multiple pancreatic cancer-associated genes.
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Objective:To investigate the reasonable approach and surgical indication for acute necrotizing pancreatitis(ANP)by analyzing the factors that affecting the mortality of ANP.Methods:One hundred and twelve patients with ANP were retrospectively divided into two groups-the dead and the survivors.Some parameters were analyzed statistically to reveal what's the reason for death.Results:The average age,sex ratio and onset of illness were similar between two groups.And the ratio of early shock,early adult respiratory distress syndrome (ARDS),high temperature,leukocytosis and high blood glucose between two groups were also similar between two groups(P＞0.05,respectively).The important factors that affecting the mortality were:①severity of pancreatic necrosis,②improper surgical approach,③incorrect surgical indication.Conclusion:The patiets with mild or moderate ANP should mainly receive conservative treatment for 48～72 hours.The early shock and ARDS should be redressed before surgical intervention.If the operation is unavoidable,the swelling pancreas should be dissected fully,which will provide sufficient drainage after operation,and duodenostomy should be performed during operation.
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Objective To investigate the expression of neurokinin 1 receptor (NK 1R) in the biopsy specimens of colonic mucosa in patients with ulcerative colitis (UC). The relation between the expression of NK 1R and clinical severity of UC was also evaluated. Methods UC tissues were obtained from 38 patients (23 males and 15 females) undergoing colonoscopy. Normal human colon tissues were obtained from 15 irritable bowel syndrome (IBS) patients (8 males and 7 females). Reverse transcription polymerase chain reaction (RT PCR), western blot and immunohistochemistry were used to determine the expression of NK 1R. Results NK 1R mRNA expression level was enhanced in mucosae of UC compared with that of IBS. The overexpression of NK 1R was related to the severity of UC. Western blot analysis revealed that the overexpression of NK 1R protein level existed in UC. Immunohistochemistry showed moderate to strong NK 1R immunoreactivity which was localized in mononuclear cells and lymphoid follicles of lamina propria and in the crypt surface, epithelium, lymphoid follicles, submucosal arterioles and myenteric plexus neurones in the colonic mucosa of UC. Conclusions In patients with UC, the overexpression of NK 1R may be involved in the pathophysiological change in the disease and may contribute to the disruption of neuropeptide loop balance and deterioration of clinical condition.
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Objective To investigate the expression of neurokinin 3 receptor (NK 3R) in normal pancreas and pancreatic cancer, and to study the localization of this receptor in pancreatic cancer Methods The expression of NK 3R mRNA was investigated using reverse transcription polymerase chain reaction (RT PCR) in normal pancreas and pancreatic cancer tissues The protein level of NK 3R was investigated by Western blot Immunohistochemistry was used to localize expression site of NK 3R Results NK 3R mRNA was overexpressed in pancreatic cancer tissue when compared with normal pancreas NK 3R protein was 1 1?0 5 in normal pancreas and 14 6?3 6 in pancreatic cancer, P
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AIM: To investigate the expression of neurok inin-2 receptor (NK-2R) in normal pancreas and chronic pancreatitis (CP) tissues. The relation of expres sion of NK-2R with pain in CP was also evaluated. METHODS: CP tis sues were ob tained from 18 men and 7 women undergoing pancreatic head resection as a result of CP. Normal human pancreatic tissues were obtained from 20 patients (11 males/ 9 females). Real-time quantitative reverse transcription polymerase chain reacti on was used to determine the mRNA expression of NK-2R, Western blot analysis was used to determine its protein expression level, and immunohistochemistry was us ed to localize expression site of NK-2R protein. NK-2R mRNA level and pain were also analysed whether correlation exists. RESULTS: NK-2R mRNA and p rotein level were enhanced expressed in CP tissue samples compared with normal pancreas. Ove rexpression of NK-2R was related to the intensity ( r=0 59, P
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AIM: To study the expression and the role of neurokinin-1 receptor(NK-1R) and neurokinin-2 receptor (NK-2R) in distal ileum during acute necrotizing pancreatitis(ANP).METHODS: 130 adult Sprague-Dawley rats were divided into 2 groups: the rats in ANP group( n=80 ) were induced by the retrograde intraductal infusion of 5% sodium taurocholate. The rats in sham operation control group ( n=50 ) received laparotomy only. Reverse transcription polymerase chain reaction (RT-PCR) was used to determine the mRNA expression of NK-1R and NK-2R, Western blot was used to investigate expression level of NK-1R.RESULTS: NK-1R and NK-2R mRNA levels were enhanced in the distal ileum of ANP rats compared with controls. Western blot revealed stronger NK-1R immunoreactivity exited in intestinal mucosa in ANP rats. The overexpression of NK-1R was associated with mucosal pathological score( r=0.77,P
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Objective To investigate the role of substance P (SP) and its neurokinin-1 receptor (NK-1R) in the pathophysiologic process of Crohn′s disease. Methods In 23 surgical patients of Crohn′s disease and 24 healthy controls, reverse transcription polymerase chain reaction (RT-PCR) was used to determine the mRNA expression of NK-1R, Western blot analysis was used to determine NK-1R protein expression levels, and immunohistochemistry was used to localize expression of NK-1R. Results Compared with normal gut NK-1R mRNA and NK-1R protein in Crohn′s disease were overexpressed. In Crohn′s disease moderate to strong intestinal NK-1R immunoreactivity was found in the lamina propria mononuclear cells, lymphoid follicles, and the surface and crypt epithelium, lymphoid follicles, submucosal vessels, smooth muscle and myoenteric plexus neurones. Conclusions In cases of Crohn′s disease, overexpression of NK-1R may disturb neuropeptides loop balance, and may be involved in the pathophysiological change in this disease.
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Objective To investigate the mechanism by which pancreatic cancer cells(PCC) acquire drug resistance against 5-FU and gemcitabine. Methods The cytotoxic effects of 5-FU and gemcitabine on PCC ( Panc-1, Mia-Paca-2 andCapan-1) were assessed by using Sulforhodamine B and the expression of anti-apoptotic genes of the Bcl-x L and mcl-1 were analyzed by RNase protection assay and Western blot. Results5-FU and gemcitabine effect cytotoxicity towards PCC. After repeated treatment with 5-FU, the IC 50 values in Capan-1 cells increased by 2.1 fold (P