OBJECTIVE:To establish a method for determining the plasma concentration of paeoniflorin and phillyrin and phar-macokinetic study before and after intragastric administration of Qianliean granules. METHODS:LC-MS/MS method was adopted. The column was Waters C18 with mobile phase consisted of acetonitrile(A)-2 mmol/L ammonium acetate(containing 0.05% formic acid)(B)(0-9 min:15%A→50%A;9-11 min:50%A→90%A;11-17 min:90%A;17-19 min:90%A→15%A;19-20 min:15%A),at the flow rate of 0.6 ml/min;column temperature was 35 ℃ and the volume was 20 μl;quantitative ions were paeoniflorin m/z 525.2 → m/z 449.0,phillyrin m/z 552.3 → m/z 355.3. 7 SD male rats were docked to collect blood 0.5 ml from angular vein 0.25,0.5,0.75,1,1.5,2,3,4,6,8,10,12,24 h after administration Qianliean granule solution 1 g(medicinal materials)/kg to determine the blood concentration of drugs. DAS 2.1.1 software was employed to calculate pharmacokinetic parameters. RE-SULTS:The linear range of paeoniflorin and phillyri were 5.0-2500.0 μg/L(r=0.9979)and 2.0-2000.0 μg/L(r=0.9982),re-spectively;RSD of precision test was less than 5.5%(n=5);the method recovery were 96.0%-104.0% and 92.0%-107.0%,the extration recovery were 71.4%-83.5% and 81.5%-92.3% and RSD of stability test was less than 5.0%(n=3). The pharmacokinet-ic parameters of paeoniflorin and phillyrin were as follows as t1/2 of (2.206 ± 0.631) and (1.355 ± 0.317) h;cmax of (1504.069 ± 620.885) and (79.043 ± 15.568)μg/L;tmax of (1.000 ± 0.250) and (1.214 ± 0.267) h;AUC0-24 h of (4897.645 ± 2207.577) and (263.475±54.795)μg·h/L;CL of(5.025±2.773)and(76.253±13.986)L/(h·kg). CONCLUSIONS:The method is highly sensi-tive,exclusive,simple,accurate and reliable,and can be applied to study the pharmacokinetic characteristics of paeoniflorin and phillyrin in rats in vivo.

Objective To explore the changes of plasma orexin-A level before and after operation in obstructive sleep apnea-hypopnea syndrome(OSAHS) children and its effect on their behavior performance.Methods 120 cases OSAHS children performed tonsillectomy and / or adenoidectomy and 30 cases normal children as control group.According to the AHI index,the OSAHS group was divided into mild group (5 times/h ≤ AHI ＜ 20times/h,n=13),moderate group (20/h ≤ AHI ＜40/h,n=76),and severe group (AHI ≥ 40 times / h,n=31).And at the same time,according to the sensory integration ratings,OSAHS group was divided into normal group(n =30),mildly abnormal group (n =47),moderately abnormal group (n =28),severely abnormal group (n =15).Before operation and 6 months after operation,plasma orexin-A levels and children's sensory integration were measured.Results Plasma orexin-A level of the OSAHS group ((0.41 ± 0.06) μg/ml) was significantly higher compared with the control group((0.31±0.04) μg/ml) (P＜0.01).In orexin-A level of different AHI groups before and after operation(mild group:(0.33±0.02) μg/ml vs (0.28± 0.03) μg/ml,moderate group:(0.39±0.04) μg/ml vs (0.29±0.03) μg/ml,severe group:(0.49±0.04) μg/ml vs (0.32± 0.02) μg/ml),there had significant differences (P＜0.01).In OSAHS children,AHI index had positive correlation with preoperative plasma orexin-A level (r=0.803,P＜0.01).There was a significant negative correlation between sensory integration scores and plasma orexinA level(r=-0.812,P＜0.01).Conclusions Plasma orexin-A level of OSAHS children is closely related to the severity of OSAHS and the changes of their behavioral ability.And it may become a diagnostic plasma marker of OSAHS children.

OBJECTIVE:To study the influence of butylphthalide on the pharmacokinetics of aspirin in rats. METHODS:20 rats were randomly divided into control group(vegetable oil 0.4 ml/rat+aspirin 10 mg/kg)and trial group(butylphthalide 80 mg/kg+aspirin 10 mg/kg) intragastrically,once a day,for consecutive 10 days. Blood samples were collected before the last medication and 10,20,40,60,120,240,360,480,600 and 720 min after medication,0.2 ml each time. The blood concentration of drugs was determined by HPLC,and pharmacokinetics parameters were calculated by DAS 2.0 software. RESULTS:Main pharmacokinet-ic parameters of aspirin in control group vs. trial group were as follows as cmax of (28.68 ± 6.08) vs. (29.33 ± 4.25)μg/ml;t1/2 of (2.48±0.67)vs.(1.60±0.36)h;AUC0-720 min of(188.71±24.29)vs.(140.31±15.08)μg·h/ml;CL/F of(0.05±0.01)vs.(0.07± 0.01)L/(h·kg);there were significant differences in t1/2,AUC0-720 min and CL/F(P<0.05). CONCLUSIONS:Butylphthalide has no significant effect on the absorption and distribution of aspirin in rats,but can strengthen its metabolism and elimination.

OBJECTIVETo evaluate the effect of new regional cooperative rescue model on the first medical contact-to-balloon time and outcome in patients with ST-elevation myocardial infarction.

METHODPatients with acute myocardial infraction (AMI) and onset time within 24 hours transferred from other hospitals to our clinic and underwent emergent percutaneous coronary intervention (PCI) between January 2010 and January 2013 were included in this study. Patients were divided into two groups: regional cooperative treatment group (n = 230) and control group (n = 168) according to whether the first contact clinic belongs to the regional cooperative rescue model or not. The first medical contact to balloon (FMC-to-B) time, door to balloon (D-to-B) time, referral time, cardiac function, mean cost, days of hospitalization, and major adverse cardiac event (MACE) during the 6 months follow up were compared.

RESULTSMean FMC-to-B time, D-to-B time and referral time were significantly decreased from (212 ± 37), (107 ± 18), (103 ± 23) min (control group) to (98 ± 23), (25 ± 7), (62 ± 12) min respectively in regional cooperative treatment group. Mean medical cost (42 221 (23 184, 77 768) RMB vs. 49 654 (25 126, 122 433) RMB) and days of hospitalization (7 (5, 13) days vs. 10 (6, 20) days) were also significantly lower in regional cooperative treatment group than in control group. At 6 months follow up, LVEF was significantly higher(54.9% ± 8.6% vs. 48.9% ± 9.1%, P = 0.01), LVEDD ((48.9 ± 5.7)mm vs.(51.4 ± 6.0) mm, P < 0.01) as well as MACE rate (7.4% (17/230) vs. 17.9% (30/168) , P < 0.05) were significantly lower in regional cooperative treatment group than in control group.

CONCLUSIONThe regional cooperative rescue model can decrease the FMC-to-B time, improve cardiac function, and reduce both patients' financial burden and MACE in patients with acute myocardial infarction.

Angioplasty, Balloon, Coronary ; Hospitalization ; Humans ; Myocardial Infarction ; therapy ; Percutaneous Coronary Intervention ; Regional Health Planning ; Time Factors

OBJECTIVETo observe whether CD137 signaling could affect the nuclear factor of activated T cells c1 (NFATc1) expression through nuclear factor-κB (NF-κB) pathway in mice aortic vascular smooth muscle cells (VSMCs).

METHODSAdherence methods for tissues explants were used for primary culture of mouse aortic VSMCs. The mRNA expression of CD137 and NFATc1 was detected by real-time quantitative PCR (RT-qPCR). The VSMCs protein expression of IκB-α, NF-κB p65, phospo-p65 and NFATc1 was determined by Western blot. The level of CD137 was measured by Flow Cytometry (FCM).

RESULTS(1) The mRNA and protein expression of CD137 in VSMCs was significantly upregulated at 24 h after co-culture with TNF-α (10 ng/ml, all P < 0.05). (2) Compared with the control group, the level of p-NF-κB p65 in cytoplasm and nucleus was significantly increased (8.34 ± 0.28 vs. 1, P < 0.05, and 2.64 ± 0.42 vs. 1, P < 0.05) while the level of IκB-α was reduced (1 vs. 2.70 ± 0.28, P < 0.05) after co-treatment with agonist-CD137 mAb, above effects were partly blocked by adding specific NF-κB inhibitor PDTC (30 µmol/L: 1.15 ± 0.14 vs. 8.34 ± 0.28, P < 0.05, and 2.09 ± 0.12 vs. 2.64 ± 0.42, P < 0.05, and 1.78 ± 0.74 vs. 1, P < 0.05). (3) The mRNA (2.07 ± 0.09 vs. 1, P < 0.05) and protein (1.75 ± 0.07 vs. 1, P < 0.05) expression of NFATc1 was significantly upregulated by agonist CD137mAb compared with the control group, and these effects could be reversed by PDTC (1.15 ± 0.07 vs. 2.07 ± 0.09, P < 0.05, and 0.90 ± 0.11 vs. 1.75 ± 0.07, P < 0.05).

CONCLUSIONCD137 signaling could affect the NFATc1expression in VSMCs through NF-kappaB pathway.

Animals ; Cells, Cultured ; I-kappa B Proteins ; Mice ; Muscle, Smooth, Vascular ; metabolism ; Myocytes, Smooth Muscle ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; NFATC Transcription Factors ; metabolism ; RNA, Messenger ; Signal Transduction ; Tumor Necrosis Factor Receptor Superfamily, Member 9 ; metabolism ; Tumor Necrosis Factor-alpha ; Up-Regulation

BACKGROUNDThe hippocampus and amygdala exhibit structural and functional alterations in patients with depression. The objective of this study was to investigate the structural and functional relationships between these core regions.

METHODSBased on the severity of their condition, 60 patients and 20 healthy controls were equally divided into four groups (mild group, moderate group, major group and health controls group), scanned by T1-MR, functional magnetic resonance imaging (fMRI), and susceptibility weighted imaging (SWI). Structural image, BOLD image, and SWI image were collected for processing and analysis. The characteristics of the depression and controls were checked by analysis of variance test, and the difference between groups was checked by Dunnett's test.

RESULTSThe volume of hippocampus and amygdala varied with the severity of the condition. The signal obtained under the stimulation of negative events was linearly decreased in the mild, moderate and major groups revealed by fMRI. The length and diameter of the lateral ventricle vein was reduced in the mild group, whereas the number of branches increased. In the moderate and major groups, the reduction in the length, diameter and increase in the number of branches of the lateral ventricle vein were greater.

CONCLUSIONThe alterations of the volume, fMRI, and cerebral veins in these core regions may account for the causal relationship between structure and function.

Adult ; Amygdala ; pathology ; Depression ; pathology ; Female ; Hippocampus ; pathology ; Humans ; Magnetic Resonance Spectroscopy ; Male

Objective　Acinetobacter baumannii (A. baumannii) is a commonly infective bacterium in the hospital. This study aims to analyze its molecular epidemiological characteristics, detect the carrying rate of efflux pump and regulatory protein genes, and investigate the effects of tigecycline on the efflux pump and expression of regulatory protein genes.　Methods　A total of 183 A. baumannii strains were collected from inpatients of the affiliated hospital of Jiangsu University from May 2017 to March 2019. They were divided into an antimicrobial-resistant group (one or more antimicrobial-resistant strains, 139 strains) and a sensitive group (the drugs in the drug sensitivity test were all non-resistant strains, 44 strains). Repeated sequence PCR was used for homology analysis of the strains, and pulse-field gel electrophoresis (PFGE) was used as the gold standard for homology analysis to verify and compare some strains. PCR was used to detect the occurrence of drug resistance-related genes. Based on homology analysis, efflux pump carrying rate detection and antibiotics sensitivity test results, 6 clinical strains carrying all efflux pump genes but different resistance phenotypes were selected as experimental strains, including sensitive strains (SAB), the multidrug resistance strain (MDRAB) and the extensively drug-resistant strain (XDRAB). All strains were induced in vitro with the minimum inhibitory concentration (MIC) of tigecycline. The induced strains were categorized as induction group, and the same strains cultured in LB agar without tigecycline was used as a control group. MIC was used to analyze the tigecycline susceptibility, and RT-qPCR was used to detect the gene expression of efflux pumps, such as TetB, AbaQ and regulatory proteins (AdeS and BaeS), in drug-resistant strains.　Results　Homology analysis showed that there were 45 clonal groups in the detected clinical isolates, with no obvious outbreak of epidemic clonal groups. Efflux pumps and regulatory proteins were widely distributed in the clinical isolates, and the expression of AdeB, TetB, AbeS, AdeS in MDRAB and XDRAB is significantly higher than that insensitive group SAB. Continuous in vitro induction with tigecycline could increase the antimicrobial resistance of some clinical strains and even significantly increase the expression levels of efflux pumps and regulatory proteins.　Conclusion　A. baumannii is widely distributed in the clinic, and efflux pumps and regulatory proteins might play an important role in drug resistance process. The unreasonable use of tigecycline could enhance the tolerance of A. baumannii by up-regulating the expression of some bacterial efflux pumps.

Objective: To evaluate the changes of left ventricular function in patients with ST segment elevation myocardial infarction (STEMI) before PCI and within 24 hours after PCI by layer-specific strain, and to explore the value of this new assessment method for quantitative monitoring on the myocardial function in STEMI patients. Methods: A total of 40 patients with acute anterior wall myocardial infarction, who underwent PCI in Affiliated Hospital of Jiangsu University during July 2017 to July 2018, were included in this prospective cohort study. According to the symptom to balloon time (STB), the patients were divided into STB ≤6 hours group (26 cases) and STB 6-12 hours group (14 cases). Echocardiography was performed before, immediately, 3 hours and 24 hours after PCI. Echocardiographic indexes including endocardial myocardial longitudinal strain (LS-endo), 18-segment full-thickness myocardial longitudinal strain (LS) of left ventricle and left ventricular global longitudinal strain (GLS) were measured. The mean LS-endo and LS values of myocardial segments in infarcted area (IALS-endo, IALS) and the mean LS-endo and LS values of myocardial segments in non-infarcted area (NIALS-endo, NIALS) were calculated. Results: There were 34 males and 6 females in this cohort and age was (62±10) years. In STB≤6 hours group, the IALS-endo value ((13.7±4.9)% vs. (10.0±2.7)%, P<0.05) and NIALS-endo value ((17.0±2.9)% vs. (14.6±2.9)%, P<0.05) were significantly higher at 24 hours after PCI than those before PCI. In the group of STB 6-12 hours, IALS-endo decreased immediately after PCI ((6.7±3.3)% vs. (11.9±6.5)%, P<0.05), and there was a rising trend at 3 hours after PCI (P>0.05). At 24 hours after PCI, the index was higher than that immediately after PCI ((13.6±8.4)% vs. (6.7±3.3)%, P<0.05). The NIALS-endo value was significantly higher at 24 hours after PCI than that before PCI ((17.1±2.1)% vs. (14.5±3.2)%, P<0.05). In the STB 6-12 hours group, the decrease rate of IALS-endo immediately after PCI was higher than that in the STB ≤6 hours group (93% (13/14) vs. 35% (9/26), P<0.001). In STB ≤6 hours group, the NIALS value at 24 hours after PCI was higher than that before PCI (P<0.05), and there was no significant difference in IALS, NIALS and GLS at other time points (P>0.05). Conclusions: Layered LS is superior to full-thickness LS and GLS in evaluating left ventricular function in STEMI patients. LS measured by echocardiography can continuously and quantitatively evaluate the changes of left ventricular myocardial function in STEMI patients before and after PCI.
Echocardiography ; Female ; Humans ; Male ; Percutaneous Coronary Intervention ; Prospective Studies ; ST Elevation Myocardial Infarction/surgery* ; Ventricular Function, Left

OBJECTIVETo investigate if miR-145a-5p participates the modulation process of CD137 signaling on the expression of nuclear factor of activated T cells c1 (NFATc1) in ApoE(-)/(-) mice.

METHODSAtherosclerotic plaque model was produced by perivascular carotid collar placement in ApoE(-)/(-) mice. After surgery, the mice were randomly divided into the following groups: CD137 activated group (CD137 group, n = 6), CD137 inhibited group (anti-CD137 group, n = 6) and control group (n = 6). The mRNA expression of miR-145a-5p in plaque and cells was measured by real-time quantitative PCR (RT-PCR). Immunofluorescence was used to observe the distribution of NFATc1 in plaque and the expression of NFATc1 at mRNA and protein levels were detected by qRT-PCR, Western blot, respectively. The mouse vascular smooth muscle cells (VSMCs) were isolated and transfected with miR-145a-5p mimics or inhibitors by Lipofectamine. The eukaryotic expression vector and luciferase vector including p3xFLAG-NFATc1, p3xFLAG-NFATc1-3'UTR, psicheck2-NFATc1, psicheck2-NFATc1-Mut were constructed through molecular cloning and homologous recombination techniques, 293T cells were transfected with the miR-145a-5p mimics or inhibitors and the protein level and fluorescence intensity were then measured, respectively.

RESULTSIn vivo or in vitro, the level of miR-145a-5p was significantly decreased (0.21 ± 0.06 vs. 1.00 ± 0.00, P < 0.05, 0.22 ± 0.07 vs. 0.50 ± 0.12, P < 0.05) while the opposite effects were observed in anti-CD137 group. NFATc1 expression was decreased or increased in VSMCs transfected with miR-145a-5p mimics or inhibitors, respectively (all P < 0.05). miR-145a-5p mimics decreased the expression of p3xFLAG-NFATc1-3'UTR and the fluorescence intensity (0.56 ± 0.08 vs. 1.00 ± 0.00, P < 0.05).

CONCLUSIONCD137 signaling participates the regulation process on the expression of NFATc1 through miR-145a-5p in ApoE(-)/(-) mice.

Animals ; Apolipoproteins E ; Mice ; Mice, Knockout ; MicroRNAs ; Muscle, Smooth, Vascular ; Myocytes, Smooth Muscle ; NFATC Transcription Factors ; Plaque, Atherosclerotic ; RNA, Messenger ; Signal Transduction ; T-Lymphocytes ; Transfection ; Tumor Necrosis Factor Receptor Superfamily, Member 9

OBJECTIVETo investigate the effects of CD137-CD137L interaction on the nuclear factor of activated T cells c1 (NFATc1) in apolipoprotein E knockout (ApoE(-/-)) mice.

METHODSAtherosclerotic plaque model was produced by rapid perivascular carotid collar placement in ApoE(-/-) mice. In vivo, the expression of NFATc1 in mice plaque and lymphocytes was detected by immunohistochemical and flow cytometry, respectively. In vitro, the NFATc1 mRNA and protein expressions in cultured lymphocytes of ApoE(-/-) mice were measured by RT-PCR and flow cytometry, respectively.

RESULTSIn vivo, after stimulating CD137-CD137L signal pathway, the expression of NFATc1 was significantly upregulated in the atherosclerotic plaques and lymphocytes. In vitro, the mRNA and protein expressions of NFATc1 in cultured leukocytes of ApoE(-/-) mice were also significantly increased, the maximal effect appeared post 20 µg/ml anti-CD137 mAb-stimulation and reached maximum at 24 h at any concentrations. Anti-CD137L mAb significantly downregulated the mRNA and protein expressions of NFATc1 in lymphocytes of ApoE(-/-) mice, maximal effect appeared at 20 µg/ml anti-CD137L mAb and reached minimum at 24 h.

CONCLUSIONCD137-CD137L interactions can modulate the expression of NFATc1 in this ApoE(-/-) mice atherosclerotic plaque model.

4-1BB Ligand ; metabolism ; Animals ; Apolipoproteins E ; genetics ; Disease Models, Animal ; Mice ; Mice, Knockout ; NFATC Transcription Factors ; metabolism ; Plaque, Atherosclerotic ; metabolism ; RNA, Messenger ; genetics ; Tumor Necrosis Factor Receptor Superfamily, Member 9 ; metabolism