ABSTRACT
OBJECTIVE To investigate the improvement effect and mechanism of calycosin （CA） on acute inflammatory injury secondary to intracerebral hemorrhage. METHODS Male C57BL/6 mice were injected with type Ⅶ collagenase into the basal ganglia to establish an intracerebral hemorrhage model， which were divided into sham-operation group（phosphate buffered saline instead of collagenase）， model group， and different CA dose groups（15，30，60，120 mg/kg）. Based on the modified neurological severity score （mNSS） to screen the intervention doses， the volume of intracerebral hemorrhage， brain water content， the expressions of ionized calcium-binding adaptor molecule 1 （Iba1） in brain tissue， Toll-like receptor 4 （TLR4） and its downstream inflammatory factors [tumor necrosis factor-α （TNF-α）， inducible nitric-oxide synthase （iNOS）， interleukin-1β （IL- 1β）] in brain tissue， and the apoptosis of cells in brain tissue were detected. Primary microglia were cultured in vitro， and the expressions of TLR4 and its downstream inflammatory factors were detected. Primary neurons and primary microglia were co- cultured in vitro， and the apoptosis of neurons was detected. RESULTS The doses of 30 mg/kg and 60 mg/kg were selected as intervention doses of CA for subsequent experiments. Compared with the sham-operation group， the mice in the model group had cerebral hemorrhage， the volume of cerebral hemorrhage and brain water content were significantly increased （P＜0.05）； the positive expression rate of Iba1 protein in brain tissue was significantly increased， and the relative expression levels of TLR4， TNF-α， IL-1β and iNOS protein in brain tissue were up-regulated significantly. The apoptosis rate also increased significantly （P＜0.05）. Compared with model group， the above indexes of the mice in the 30 and 60 mg/kg CA groups were significantly improved （P＜0.05）. CA significantlyreduced the relative expression levels of TLR4 and its downstream inflammatory factors in microglia， and reduced the apoptosis of neurons in the co-culture system of primary neurons and primary microglia （P＜0.05）. CONCLUSIONS CA can exert a protective effect on the brain， which may be related to relieving the secondary acute inflammatory injury after intracerebral hemorrhage by inhibiting TLR4-mediated inflammatory response.
ABSTRACT
OBJECTIVE：To provide reference for the participation of clinical pharm acists in outpatient antibiotics management so as to promote the rational use of antibiotics in outpatient department. METHODS ：The pharmacist reset medication rules ， classified irrational medication levels ，formulated warning contents ，and established a diagnosis database of mild to moderate non- complex infections ，according to the problems found in the post review of outpatient antibiotics prescription through sorting out the previous rules of antibiotics use in the Yiyao Rational Drug Use Management Software （called“review software ”for short ）in our hospital. The special pre-review was performed by using combination of system review and manual review. Pharmacists used the review saftware to count the data of outpatient antibiotics prescriptions before （the first quarter of 2020）and after （the second ， third and fourth quarters of 2020）the implementation of special pre-review. The post review for antibiotics prescriptions in Dec. 2019（before the implementation of special pre-review ）and Dec. 2020（after implementation of special pre-review ）were performed by pharmacists to evaluate the impact of the implementation of special pre-review on promoting rational use of outpatient antibitics. RESULTS：Comared with before the implementation of special pre-review in the first quarter of 2020，the prompt rate of the review software for antibiotics prescriptions was decreased ，and the interception rate was increased after the implementation of special pre-review in the second quarter of 2020（P＜0.05）. Compared with before the implementation of special pre-review in Dec. 2019，the proportion of outpatient antibiotics prescriptions in all outpatient prescriptions ，the ratio of bi-antibiotics prescriptions in all antibiotics prescriptions in the same month ，the ratio of prescriptions with unsuitable indications in all antibiotics prescriptions in the same month were all decreased E-mail：phdloveyou@163.com significantly （P＜0.01）， while the ratio of prescriptions with rational antibiotics use in all antibiotics prescriptions in the same month was increased significantly （P＜0.01）. CONCLUSIONS：Pharmacists establishing a rule for rational use of antibiotics in outpatient department b ased o n the hospital ’s situation，implementing a special pre-review for antibiotics prescriptions with the help of review software ，can promote the rational use of antibiotics in outpatient department.
ABSTRACT
Objective To investigate the effect of paeoniflorin on epithelial-mesenchymal transition (EMT),migration and invasion of human glioma U87 cells and its underlying molecular mechanism.Methods Glioma U87 cells were treated with different dose of paeoniflorin.CCK-8 was used to measure the survival rate of glioma cells.Glioma U87 cells were infected with carrying transforming growth factor-β(TGF-β)lentivirus to establish the stable overexpression TGF-β cell line.Wound healing assay and transwell assay were performed to measure cell migration and invasion ability,respectively.Western blot was applied to examined the expression of related proteins.Results Treatment with proliferation of 5 mmol/L and 10 mmol/L had no significant effect on cell survival of U87 cells ((96.00± 3.61) ％,(92.33± 4.04) ％;P5 =0.593,P10 =0.284).But when the dose was up to 15 mmol/L,the survival rate of U87 cells decreased to(75.67±4.58)％,P=0.008.In addition,paeoniflorin inhibited the migration ability of U87 cells and the wound healing rate was(68.20±4.39) ％,(37.70±7.01) ％ when incubated with 5 mmol/L and 10 mmol/L.Similarly,paeoniflorin suppressed invasion ability of U87 cells and the average number of infiltrated cells was (237.67±24.00),(130.67± 32.65),(59.67± 18.15) respectively (P5 =0.002,P10 ＜ 0.01).What's more,paeoniflorin down-regulated the expression of MMP2,MMP9 and epithelial-mesenchymal transition-related proteins (P＜0.05).Overexpression of TGF-β reversed the effect of paeoniflorin on migration,invasion and epithelial-mesenchymal transition of glioma U87 cells.Conclusion Paeoniflorin suppress TGF-β-induced migration,invasion and epithelial-mesenchymal transition in glioma U87 cells.
ABSTRACT
Objective To investigate the effect of paeoniflorin on epithelial-mesenchymal transition (EMT),migration and invasion of human glioma U87 cells and its underlying molecular mechanism.Methods Glioma U87 cells were treated with different dose of paeoniflorin.CCK-8 was used to measure the survival rate of glioma cells.Glioma U87 cells were infected with carrying transforming growth factor-β(TGF-β)lentivirus to establish the stable overexpression TGF-β cell line.Wound healing assay and transwell assay were performed to measure cell migration and invasion ability,respectively.Western blot was applied to examined the expression of related proteins.Results Treatment with proliferation of 5 mmol/L and 10 mmol/L had no significant effect on cell survival of U87 cells ((96.00± 3.61) ％,(92.33± 4.04) ％;P5 =0.593,P10 =0.284).But when the dose was up to 15 mmol/L,the survival rate of U87 cells decreased to(75.67±4.58)％,P=0.008.In addition,paeoniflorin inhibited the migration ability of U87 cells and the wound healing rate was(68.20±4.39) ％,(37.70±7.01) ％ when incubated with 5 mmol/L and 10 mmol/L.Similarly,paeoniflorin suppressed invasion ability of U87 cells and the average number of infiltrated cells was (237.67±24.00),(130.67± 32.65),(59.67± 18.15) respectively (P5 =0.002,P10 ＜ 0.01).What's more,paeoniflorin down-regulated the expression of MMP2,MMP9 and epithelial-mesenchymal transition-related proteins (P＜0.05).Overexpression of TGF-β reversed the effect of paeoniflorin on migration,invasion and epithelial-mesenchymal transition of glioma U87 cells.Conclusion Paeoniflorin suppress TGF-β-induced migration,invasion and epithelial-mesenchymal transition in glioma U87 cells.