ABSTRACT
ObjectiveThis study aims to investigate the mechanism in which Celastrus orbiculatus extract （COE） affects the proliferation and differentiation of gastric organoids and the expression of Lgr5 and thus reverses the precancerous lesions of gastric cancer （PLGC） by regulating the leucine-rich repeat containing G protein-coupled receptor 5 （Lgr5）/Wingless （Wnt）/β-catenin signaling pathway based on a gastric organoid injury model. MethodGastric organoids were established based on stem cells of the mouse gastric gland. Gastric organoid injury models were constructed by treating gastric organoids with N-methyl-N'-nitro-N-nitrosoguanidine （MNNG， 0.02 mg·L-1）. Gastric organoid injury models were randomly divided into normal group， model group （0.02 mg·L-1 MNNG）， low， medium， and high dose （5， 10， 20 mg·L-1） groups of COE， and Wnt inhibitor Dickkopf-related protein 1 （DKK1） （0.5 mg·L-1） group， and they were treated with respective agents for 24 h. The number and volume of gastric organoids under different drug concentrations were observed under a microscope. The viability of the gastric organoid injury models was detected by Methyl thiazolyl tetrazolium （MTT） assay. The morphology and pathology of gastric organoids were observed using Hematoxylin and Eosin （HE） staining. The expression levels of Lgr5， Mucin2 （MUC2）， Mucin5AC （MUC5AC）， Mucin6 （MUC6）， Wnt， and β-catenin in gastric organoids under different drug concentrations were detected by Western blot （WB）. ResultCompared with the normal group， the number， volume， and activity of gastric organoids in the model group were decreased （P<0.01）， while the expressions of Lgr5， MUC2， Wnt， and β-catenin were significantly increased （P<0.01）. The expressions of MUC5AC and MUC6 were significantly decreased （P<0.01）. Compared with the model group， the number and volume of gastric organoids in the low， medium， and high dose groups of COE were all improved （P<0.01）， and the vitality of gastric organoids was significantly enhanced （P<0.01）. The effect was the most significant at a COE concentration of 20 mg·L-1 （P<0.01）. The expressions of Lgr5 and MUC2 in the medium and high dose groups of COE were significantly reduced （P<0.01）， while the expression of MUC5AC and MUC6 were significantly increased in the low， medium， and high dose groups of COE （P<0.05， P<0.01）. Compared with the model group， Wnt inhibitors could promote the expression of MUC5AC and MUC6 in gastric organoids （P<0.05， P<0.01） and reduce the expression of MUC2， Wnt， and β-catenin. In addition， the combined use of COE at high concentrations and Wnt inhibitors could further promote this trend （P<0.01）. ConclusionCOE inhibits the Wnt/β-catenin pathway by inhibiting the expression of Lgr5， MUC2， Wnt， and β-catenin and promoting the expression of MUC5AC and MUC6， thus promoting the proliferation and differentiation of gastric organoids and reversing the PLGC process.
ABSTRACT
Objective@#To investigate the effect of Celastrus orbiculatus extract ( COE) on the growth of gastric organoids and the expression of E-cadherin in gastric epithelial cells of mice．@*Methods@#The gastric pylorus of 8-week-old C57 mice was isolated and cultured into gastric organoids．The dynamic changes of gastric organoid formation were observed under light microscope ; the intercellular structure of gastric epithelium was observed by HE staining ; the expression of epithelial-cadherin E-cadherin in gastric epithelial cells was observed by immunofluorescence staining．After passage to the third-generation ，the organoids were treated with different concentrations of COE (0，5 ，10，20 μg/ ml) ，the organoids were collected ，their numbers were counted ，their diameters were measured，their cellular activities were measured by methyl thiazolyl tetrazolium( MTT) colorimetry，and Western blot was used to detect the expression of E-cadherin in organoids after COE treatment. @*Results @#At 24 to 48 h， cystlike structures were formed and three-dimensional cell clusters with cystic gland-like central structure appeared， and gradually budding and forming gastric organoids after 72 h，suggesting that the organoids were successfully constructed．The epithelial cell marker E-cadherin was expressed in the organoid，which further confirmed the formation of organoid．Compared with the control group，the number and diameter of gastric organoids in the COE group significantly increased，cell activity was significantly enhanced (P＜0. 05) ，and the expression of E-cadherin increased with the increase of COE dose (P＜0. 01) ．@*Conclution @#Low dose COE can promote the expression of E-cadherin and the growth and formation of organoids，which may affect the repair of gastric mucosa injury.
ABSTRACT
Objective To investigatethe EC50 of different doses of dexmedetomidine on etomidate inhibited responses tolaryngeal maskinsertion in patients.Methods 88 with breast cancerfrom surgical department in Zhuji Hospital of Traditional Chinese Medical from August 2014 to August 2015were selected and randomly divided into the control group(groupA)and the experiment group(group B1,group B2 and group B3)with 22 casesin each group,respectively,intravenous pump 0.9％sodium chloride solution and DEX(dose followed by 0.3,0.6,0.9μg/kg).The next sequential intravenous infusion of etomidateafter 10 min.The EC50 and the 95％confidence interval of etomidate were determined by sequential method in each group of patients,the changes of vital signs and adverse reactionsin patients were monitored.Results Four groups of patients with LMA EC50 and 95％confidence interval of etomidate respectively:0.78(0.723～0.835)μg/mL in group A,0.66(0.612～0.711)μg/mL in group B1,0.58(0.532～0.627)μg/mL in group B2,0.46(0.416～0.521)μg/mL in group B3.Four groups of patients with laryngeal mask insertion immediately before MAP and HR were lower than the baseline value,the difference was statistically significant(P<0.05),but the LMA elevated after one min.The control group of four patients with respiratory depression,three cases of patients with bradycardia in group B3,two cases of patients with hypotension were improved after symptomatic treatment.Conclusion In a certain range,increasing the dose of dexmedetomidine reduces the effective concentration of etomidate,which inhibits the laryngeal mask placement reaction.
ABSTRACT
Objective To assess the accuracy of bispectral index (BIS) in monitoring the sedation level with dexmedetomidine combined with small-dose of sufentanil.And, to analyze the BIS value of the best opportunity for invasive manipulation with this sedation method.MethodsEighty elective operation patients, ASA Ⅰ~Ⅱ, were randomLy selected for the study.Before the start of anesthesia, small-dose of sufentanil 0.1μg/kg and dexmedetomidine were injected in vein in sequence.The loading dose of dexmedetomidine was 1.0μg/kg for 15 min and the maintenance dose was 0.3μg/kg for 5 min.The time-pint before the start of injection was recorded as T0.And, T1, T2, T3, and T4 respectively presented the infusion time of dexmedetomidine for 5min, 10min, 15min and 20min.According to these four time-points, 80 patients were randomLy divided into four groups: group T1, group T2, group T3,and group T4.The relevant HR, MAP, SpO2, BIS value, and OAA/S score were separately recorded.Analyze the correlation of BIS and OAA/S score, and the cutoff value of BIS for OAA/S score=1 was obtained by analyzing receiver operating characteristic (ROC) curve.ResultsThe compound use of dexmedetomidine and sufentanil significantly decreased HR.With this sedation method, BIS value was positively correlated with OAA/S score (r=0.95), and the best cutoff value of BIS values for OAA/S score=1 was 44.5.ConclusionThere was a good correlation between BIS and OAA/S score in monitoring sedation with dexmedetomidine combined with small-dose of sufentanil.BIS=44.5 could be regarded as a good monitoring index of invasive manipulation with this sedation method.