2.Effect of fluvastatin on expression of SGK1 and CTGF induced by aldosterone in rat mesangial cells
Cairong LI ; Fei CAI ; Rong LI ; Xinyuan ZHAO ; Juan LI
Chinese Journal of Pathophysiology 1989;0(05):-
AIM:To investigate the effect of fluvastatin on the expression of serum and glucocorticoid inducible kinase 1 ( SGK1) and connective tissue growth factor ( CTGF) induced by aldosterone ( Ald) in rat mesangial cells (GMCs). METHODS:GMCs were divided into (1) control group; (2) aldosterone group with different concentrations and times; (3) Ald (10 -7 mol/L) + spironolactone (10 -9 mol/L) group; (4) Ald (10 -7 mol/L) + LY294002 (20 ?mol/L) group; (5) Ald (10-7mol/L) +SB203580 (20 mmol/L) group; (6) the group of Ald (10-7mol/L) + fluvas-tatin at different concentrations (10-7,10-6,10-5 mol/L); (7) Ald (10 -7mol/L) + fluvastatin (10 -5mol/L) + mevalonate (10 -4 mol/L) group; (8) Ald (10 -7 mol/L) + fluvastatin (10 -5 mol/L) + FPP (farnesyl pyrophosphate,10-4 mol/L) group; (9) Ald (10 -7mol/L) + fluvastatin (10 -5 mol/L) + GGPP (geranylgerany pyrophosphate,10 -4 mol/L) group. The protein levels of SGK1 and CTGF were determined by Western blotting. The levels of fibronection (FN),monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) in the supernatants were determined by enzymelinked immunosorbant assay (ELISA). RESULTS:Aldosterone stimulated the protein expression of SGK1 and CTGF in cultured mesangial cells in a dose-dependent manner (P
3.Determination of serum acetaminophen based on the diazo reaction and its application in the evaluation of gastric emptying.
Cai-na LI ; Su-juan SUN ; Zhu-fang SHEN
Acta Pharmaceutica Sinica 2015;50(5):560-564
This study aims to establish a method to determine the serum acetaminophen concentration based on diazo reaction, and apply it in the gastric emptying evaluation. Theoretically, acetaminophen could take hydrolysis reaction in hydrochloric acid solution to produce p-aminophenol, which could then take diazo reaction resulting in a product with special absorption peak at 312 nm. Then the serum acetaminophen concentration and recovery rate were calculated according to the standard curve drawn with absorbance at 312 nm. ICR mice were given a dose of acetaminophen (500 mg x kg(-1)) by gavage and the serum acetaminophen was dynamically measured through the diazo reaction. Besides, ICR mice were subcutaneously injected with the long-acting GLP-1 analog GW002 before the gavage of acetaminophen, and serum acetaminophen concentration was measured as above to study how GW002 could influence the gastric emptying. The data showed acetaminophen ranging from 0 to 160 μg x mL(-1) could take diazo reaction with excellent linear relationship, and the regression equation was y = 0.0181 x +0.0104, R2 = 0.9997. The serum acetaminophen was also measured with good linear relationship (y = 0.0045 x + 0.0462, R = 0.9982) and the recovery rate was 97.4%-116.7%. The serum concentration of acetaminophen reached peak at about 0.5 h after gavage, and then gradually decreased. GW002 could significantly lower the serum acetaminophen concentration and make the area under the concentration-time curve (AUC) decrease by 28.4%. In conclusion, a method for the determination of serum acetaminophen based on the diazo reaction was established with good accuracy and could be used in the evaluation of gastric emptying.
Acetaminophen
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blood
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pharmacokinetics
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Aminophenols
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Animals
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Gastric Emptying
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Mice
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Mice, Inbred ICR
4.Determination of Global DNA Methylation Levels in Medicinal Plants by LC-MS/MS
Qionglin HUANG ; Juan WEN ; Zhaojin LI ; Chun CAI
China Pharmacy 2016;27(16):2210-2212,2213
OBJECTIVE:To develop a detection LC-MS/MS method for global DNA methylation in medicinal plants. METH-ODS:Genomic DNA was isolated using plant DNA extraction kit,and then hydrolyzed by 88% formic acid at 140 ℃. After dried with nitrogen,extracted DNA was dissolved again with mobile phase. LC separation was performed on HILIC column with mobile phase consisted of 7 mmol/L ammonium formate-acetonitrile (gradient elution) at flow rate of 0.3 ml/min. The analysis was con-ducted by tandem MS with positive ion electrospray ionization in multiple reaction monitoring(MRM)mode. The ratio of genomic DNA methylation in 10 commonly used medicinal plants was calculated. RESULTS:The linear ranges of Cyt and 5mC were 1-500 ng/ml(r=0.999 5)and 0.2-100 ng/ml(r=0.999 6). The relative standard deviations(RSDs)of accuracy were 1.12% and 3.68%(n=6). The RSDs of intra-day precision were 2.36% and 4.02% for Cyt and 5mC,respectively (n=5). The RSDs of inter-day precision were 1.04% and 3.54% for Cyt and 5mC,respectively (n=3). The RSDs of repeatability test were 1.53% and 3.27%for Cyt and 5mC,respectively(n=6). The recoveries of Cyt and 5mC were 98.7%-102.1% and 91.2%-103.5%. The percentages of global DNA methylation in 10 medicinal plants were ranged from 17.63% to 25.18%. CONCLUSIONS:LC-MS/MS method is simple,rapid,sensitive and precise,and can be used for the detection of global DNA methylation in medicinal plants.
5.Expression and significance of long non-coding RNA MALAT1 and interleukin 6 in premature infants with bronchopulmonary dysplasia
Hongzhuan ZHANG ; Cheng CAI ; Juan LI ; Xiaohui GONG ; Xiaoyun CHU
Chinese Journal of Applied Clinical Pediatrics 2021;36(7):533-536
Objective:To investigate the expression of long non-coding RNA MALAT1, interleukin 6(IL-6) and apoptosis induced factor(AIF) in peripheral venous blood of premature infants with bronchopulmonary dysplasia (BPD) and its clinical significance.Methods:Preterm infants admitted to the Department of Neonatology, Shanghai Children′s Hospital from January 2015 to December 2016 were enrolled.The selection criteria included gestational age (GA) ≥28 weeks and ≤32 weeks, and birth weight (BW) < 1 500 g. According to the diagnosis, they were divided into BPD group (20 cases) and control group (20 cases). The clinical data of the two groups of premature infants were collected and analyzed, and the levels of MALAT1, IL-6 and AIF in the blood of 40 premature infants were detected by real-time polymerase chain reaction (RT-PCR). T test was used to compare gestational age, birth weight, MALAT1, IL-6 and AIF between the two groups. Results:(1)There was no significant differences in sex ( χ2=1.76), gestational age ( t= 0.17) and birth weight ( t=1.25) of premature infants in BPD group, compared with the control group (all P >0.05). (2)Compared with the control group, the expression of MALAT1 in the peripheral blood of premature infants in BPD group were significantly increased (0.273 4±0.067 3 vs. 0.375 5±0.081 9, P<0.05). (3)Compared with the control group, the expression of IL-6 in the peripheral blood of premature infants in BPD group were obviously decreased (1.448 8±0.191 8 vs.4.444 6±0.165 7, P<0.05). (4)Compared with the control group, the expression of AIF in the peripheral blood of premature infants in BPD group were remarkably decreased(0.006 8±0.002 0 vs.0.004 5±0.001 9, P<0.05). Conclusions:MALAT1 and IL-6 levels of long non-coding RNA in BPD and non-BPD preterm infants are different, which may be related to the incidence of BPD.IL-6 may be a predictor of BPD, and MALAT1 may protect premature infants with BPD.
6.Comparison of different detection methods of monkey B virus antibody
Jinwen LI ; Wei TONG ; Juan CAI ; Zhiguang XIANG ; Qiang WEI
Chinese Journal of Comparative Medicine 2017;27(7):29-33
Objective Monkey B virus(BV), also known as Cercopithecine herpesvirus 1,is an important zoonotic pathogen.According to the national standard, antibodies are detected using BV as an antigen.However, the preparation of BV antigen is very stricted due to biosafety issues.Therefore, in this study, we used alternative antigens to detect the BV antibody by serological assay and verified their specifity and sensitivity.Methods A total of 135 blood samples from rhesus monkeys were tested by two ELISA method (BV and HVP2) and enzyme immunosorbent assay (EIA)method.The positive and suspicious samples were verified by immuno-fluorescence assay (IFA), Western blot and immunoblotting technique using HSV-1 gC1 purified glycoprotein as an antigen.Results The positive rates of HVP2-ELISA, BV-ELISA and HSV-1-EIA were 32.6%, 37.8% and 34.8%, respectively.Consistant result of the three detection method accounted for 91.1% (123/135), and the positive result were confirmed by IFA And WB.There were 12 suspicious samples,in which 33.3% (4/12) were verified to be positive.Conclusions Compared with BV antigen, the sensitivity and specificity of the alternative antigen HSV-1 are moe close than HVP2.Positive and suspicious samples should be verified by several method to avoid missed detection.
7.Analysis of Clinical Distribution and Drug-resistance of Pathogen in Urinary Tract Infection in Wuhan
Zhen LI ; Congrong LI ; Xuan CAI ; Juan LI ; Qing WU ; Ziqi HE
Journal of Modern Laboratory Medicine 2014;(5):135-137,140
Objective To investigate the tendency of distribution and drug-resistance of the causative organisms of urinary tract infections(UTIs)in Wuhan,and provide reliable evidence for clinical treatment.Methods Analyzed the 5 378 stains of pathogen isolated from the urine of patients in hospital.The bacteria isolates were identified with BD Phoenix-100 while can-dida isolates were identified by color plate.Results A total of 5 378 stains of pathogen had been isolated.There were 2 945 stains (54.8%)of Gram-negative bacteria,1 657 stains (30.8%)of Gram-positive bacteria,776 stains (14.4%)of fungus. The rates of Escherichiacoli resistant to penicillin were highest (>83%),and there were no carbapenem-resistant strains. There were vancomycin and linezolid-resistant Enterococcispp strains,the lowest dection rates of which were 0.3%.The de-tection rate of MRCNS was over 83%.Conclusion Escherichiacoli was the most common pathogens of urinary tract infec-tion,and theβ-lactamase inhibitor complex can be used as empirical treatment of E.coli infections.Thedetection rate of MRCNS increased,which shoud be kept a watchful eye on.
8.Comparative study between stenting and medication for vertebral artery origin stenosis
Jingjing LI ; Yiling CAI ; Li LIU ; Juan DU ; Zhen WU ; Xiangkai KONG
Chinese Journal of Cerebrovascular Diseases 2015;(1):1-6
Objective Tocomparetheefficacybetweenstentingandmedicationinpatientswith vertebralarteryoriginstenosis.Methods Theclinicaldataof82patientswithmoderatetoseverevertebral artery origin stenosis (stenosis rate >50%)from January 2011 to January 2013 were analyzed retrospectively. They were divided into either a stent+medication group (n=40)or a medication group (n=42)according to the different treatment methods. The degree of vascular stenosis,restenosis rate,incidence of cerebral ischemic events,and National Institutes of Health Stroke Scale (NIHSS)of the patients for DSA reexamination at one year were documented,and comprehensive analysis were conducted. Results (1 )44 stents were implanted in 40 patients,1 of the patients still had residual stenosis of 60% because of the stenosis plaque was harder despite twice balloon dilations. No serious perioperative complications occurred. The success rate of the operation was 97. 5%. The vascular stenosis rate of vertebral artery origins in patients of the stent+medication group was improved significantly,and decreased from 73 ± 13% to median 11%(8%,50%)at one year after stenting. (2)After 1 year,11 patients (27. 5%)had in-stent restenosis in the stent +medication group,including 2 patients (5%)had stent fracture at the same time. Four patients (9. 5%)in the medication group had complete occlusion of vertebral artery,but only 2 had corresponding clinical symptoms. (3)There was no significant difference in the NIHSS scores after the treatment between the stent+medication group and the medication group (Z=1. 678,P=0. 093). The total ischemic events in patients of the stent+medication group was 7 (17. 5%),compared with the incidence of ischemic events in 16 patients (38. 1%)in the medication group,there was significant difference (χ2 =4. 306, P=0.038).Conclusion Stentingissafeandeffectiveforpatientswithvertebralarteryoriginstenosis.It may significantly improve vertebral stenosis,and it is better than medication alone for preventing the occurrence alone of the posterior circulation ischemic events,however,the high in-stent restenosis rate for vertebral artery origin stenosis is still an important problem to be solved.
9.Progress in study of selective ERβ ligands.
Jinya CAI ; Junhao LI ; Shihui DING ; Juan ZHANG ; Guixia LIU ; Weihua LI ; Yun TANG
Acta Pharmaceutica Sinica 2015;50(6):658-67
Estrogen receptors (ERs) are members of nuclear receptors and related to several diseases such as cancer, inflammation and osteoporosis. ERs have two forms, ERα and ERβ, which have different functions and organism distributions. Compounds selectively targeting ERβ can regulate important physiological functions and avoid the side effects caused by targeting ERα. Therefore, selective ERβ ligands have received considerable research interest in recent years. In this article, different kinds of selective ERβ ligands were summarized and their structure-activity relationships were also analyzed.
10.The effect of mitochondria-targeted antioxidant peptide SS31 on oxidative damage of lens epithelial cell
Meng, CAI ; Jin, LI ; Jing, LI ; Xiao-yun, CHEN ; Juan, HUANG ; Yan, LUO
Chinese Journal of Experimental Ophthalmology 2013;31(12):1137-1141
Background Presbyopia is one of primary causes affecting the visual and life qualities of the agings,and its mechanism is associated with the oxidative damage of lens epithelial cells with ageing.SS31 is a mitochondria-targeted antioxidant peptide.To study the effect of SS31 on oxidative damage of lens epithelial cells has an important significance for the prevention and treatment of presbyopia.Objective This study was to investigate the effect of SS31 on in vitro oxidative damaged human lens epithelial cells.Methods Human lens epithelial cell line (HLEB-3) was cultured using DMEM with low glucose and 10% fetal bovine serum(FBS).The cell model of oxidative damage was established by adding 200 μmol/L tea-butyl hydropeoxide (t-BHP) into DMEM for 18 hours.The cells were divided into blank control group,t-BHP model group,10 nmol/L SS31 +t-BHP group,100 nmol/L SS31 +t-BHP group,1 μmol/L SS31 +t-BHP group,10 μmol/L SS31 +t-BHP group and 100 pμmol/L t-BHP group,and then MTT assay was used to detect the survival rate of the cells and evaluate the optimal SS31 concentration for sequential study.The cells then were divided into blank control group,t-BHP model group and 1 μmol/L SS31 +t-BHP co-culture group.The change of mitochondrial membrane potential of the cells was tested by JC-1 dye and flow cytometry.Reactive oxygen species (ROS) level in the mitochondria was determined using MitoSOX staining.Results The cell survival rate in the t-BHP model group was (53.42±2.52)%,and that in the blank control group was 100%.The cell survival rate was considerably increased in various concentrations of SS31 groups,showing a significant difference among different groups (F=58.349,P<0.01).A highest survival rate was (82.13 ±3.15) % in the 1 μmol/L SS31 +t-BHP co-culture group,which was statistically significant in comparison with the t-BHP model group (t =28.710,P<0.05).JC-1 dye and flow cytometry assay showed that the ratio between red and green fluorescence intensity was 7.07 ±0.06 in the blank control group,4.46±0.14 in the t-BHP model group and 5.76±0.26 in the 1 μmol/L SS31 +tBHP co-culture group,showing significant difference among the 3 groups (F=172.332,P<0.01).The ratios between red and green fluorescence intensity in the blank control group and 1 μmol/L SS31 +t-BHP co-culture group were higher than that in the t-BHP model (t =2.609,1.303,both at P<0.001).ROS fluorescence cells were much more in the t-BHP model group compared with blank control group and 1 μmol/L SS31 + t-BHP co-culture group.Conclusions SS31 can protect HLEB-3 cells from oxidative stress.SS31 may serve as a potential new approach to the treatment of presbyopia and other age-related diseases of lens.