The relationship between M3 cholinergic receptor agonist (carbachol) hyperstimulation-induced pancreatic acinar cellular injury and trypsinogen activation or NF-kappaB activation in rats was studied ...in vitro. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor (pefabloc), and NF-kappaB inhibitor (PDTC) in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar cells. The results showed that as compared with control group, 10(-3) mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells (P < 0.01) following the treatment with a high concentration of carbachol (10(-3) mol/L) in vitro. The addition of 10(-2) mol/L PDTC didn't result in a significant decrease in the activity of trypsin and the leakage of LDH from pancreatic acinar cells treated with a high concentration of carbachol (10(-3) mol/L) in vitro (P > 0.05). It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-kappaB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro.
R3; The relationship between M3 cholinergic receptor agonist (carbachol) hyperstimulationinduced pancreatic acinar cellular injury and trypsinogen activation or NF-κB activation in rats was studied ...in vitro. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor (pefabloc), and NF-κB inhibitor (PDTC) in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar cells. The results showed that as compared with control group, 10-3 mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells (P ＜0.01) following the treatment with a high concentration of carbachol (10-3 mol/L) in vitro. The addition of 10-2 mol/L PDTC didn't result in a significant decrease in the activity of trypsin and the leakage of LDH from pancreatic acinar cells treated with a high concentration of carbachol (10-3 mol/L) in vitro (P＞0.05). It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-κB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro.
In order to construct an expression vector carrying small hairpin （sh） RNA （shRNA） for toll-like receptor 4 mRNA and a reporter gene of enhanced green fluorescence protein （EGFP） and study the ...inhibition of cytokine release by RAW264.7 cell induced by lipopolysaccharide （LPS） stimulation through transfection and expression of shRNA targeting TLR4 gene via the RNAi mecha- nism, the reporter gene plasmid pEGFP-CI （4.7 kb） and psiRNA-hHlneo （2979 bp） were used. The HI promotor and double Bbs Ⅰ restrict endoenzyme site were cloned from plasmid psiRNA-hHlneo and reconstructed them into plasmid pEGFP-CI in the Mlu Ⅰ restrict endoenzymic site, forming plasmid pEGFP-HI/siRNA, which contained Bbs site and reporter EGFP gene. Then an oligonuclear hairpin sequence targeting TLR4 gene was designed by internet tool and inserted into the plasmid pEGFP-H 1/siRNA forming plasnlid pEGFP-H 1/TLR4-siRNA. After transfection of pEGFP-H I/TLR4-siRNA into RAW264.7 cells, tumor necrosis factor-alpha （TNF-α） release by the cells after stimulation by LPS was detected. The results showed that the constructed pEGFP-HI/TLR4-siRNA carrying hairpin RNA for TLR4 gene and reporter EGFP gene were proven to be right by restriction endonuclease analysis. The expression of EGFP gene was （50.37±8.23） % and after transfection of the plasmid pEGFP-HI/ TLR4-siRNA the level of TNF-α released by RAW264.7 cell was down regulated. It was concluded that shRNA targeting TLR4 gene could inhibit the TNF-α release by RAW264.7 cells evoked by LPS.
In order to construct an expression vector carrying small hairpin (sh) RNA (shRNA) for toll-like receptor 4 mRNA and a reporter gene of enhanced green fluorescence protein (EGFP) and study the ...inhibition of cytokine release by RAW264.7 cell induced by lipopolysaccharide (LPS) stimulation through transfection and expression of shRNA targeting TLR4 gene via the RNAi mechanism, the reporter gene plasmid pEGFP-C1 (4.7 kb) and psiRNA-hHlneo (2979 bp) were used. The H1 promotor and double Bbs I restrict endoenzyme site were cloned from plasmid psiRNA-hH1neo and reconstructed them into plasmid pEGFP-C1 in the Mlu I restrict endoenzymic site, forming plasmid pEGFP-H1/siRNA, which contained Bbs site and reporter EGFP gene. Then an oligonuclear hairpin sequence targeting TLR4 gene was designed by internet tool and inserted into the plasmid pEGFP-H1/siRNA forming plasmid pEGFP-H1/TLR4-siRNA. After transfection of pEGFP-H1/TLR4-siRNA into RAW264.7 cells, tumor necrosis factor-alpha (TNF-alpha) release by the cells after stimulation by LPS was detected. The results showed that the constructed pEGFP-H1/TLR4-siRNA carrying hairpin RNA for TLR4 gene and reporter EGFP gene were proven to be right by restriction endonuclease analysis. The expression of EGFP gene was (50.37+/-8.23) % and after transfection of the plasmid pEGFP-H1/ TLR4-siRNA the level of TNF-alpha released by RAW264.7 cell was down regulated. It was concluded that shRNA targeting TLR4 gene could inhibit the TNF-alpha release by RAW264.7 cells evoked by LPS.
To study the protein expression of endotoxin, interleukin 1beta (IL-1beta) and regulated upon activation, normal T expressed and secreted (RANTES) in effusions of secretory otitis media and their ...roles in the pathogenesis of secretory otitis media.
Seventy-two effusion samples were taken from 53 patients with secretory otitis media by tympanocentesis. After swabs they were taken for bacteria culture. Limulus amebocyte lysate (LAL) test was used to quantify the content of endotoxin, radioimmunoassay to analyze the level of IL-1beta and enzyme-linked immunosorbent assay (ELISA) to detect the concentration of RANTES.
Endotoxin, IL-1beta and RANTES were detectable in 80.9%, 77.8% and 70.8% of middle ear effusion, with mean levels of (35.2 +/- 51.6) EU/ml, (1.10 +/- 0.84) microg/L and (0.52 +/- 0.43) microg/L respectively. All of them showed higher concentration in the mucoid-type effusions than those in the serous-type effusions (P<0.05). Higher levels of the endotoxin and RANTES (P<0.05) were found in longer cou
To study the influence of different managements of middle turbinate on nasal airway resistance(NAR) and nasal airflow sensation.
Patients were divided into three groups: total resection group (32 ...patients/sides), partial resection group (37 patients/sides) and total reservation group (34 patients/sides). The NAR and nasal airflow sensation were measured with anterior rhinomanometer and visual analogue scale test (VAS) respectively.
The NAR and VAS scores decreased significantly after operation (P < 0.01). But there was no significant difference between these three groups(P > 0.05).
The different managements of middle turbinate have no significant influence on NAR and nasal airflow sensation.
