Animals ; Cardiovascular Diseases ; genetics ; pathology ; Cell Proliferation ; Genes ; Microfilament Proteins ; genetics ; Muscle Proteins ; genetics ; Muscle, Smooth, Vascular ; pathology ; Rats

AIM:To establish an HPLC method for determining the contents of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd. METHODS:The samples were separated on an Alltima C 18 (250 mm? 4.6 mm,5 ?m) column with the mobile phase of MeOH(A)-0.5% glacial acetic acid solution;gradient elution(0～15 min,30%～60% A;15～30 min,60%～60% A).Flow rate was 1.0 mL/min. The detection wavelength was set at 350 nm.Column temperature was at 30 ℃. RESULTS:The contents of caffeic acid,quercetin and campherenol were 14.218～23.695 ?g/g,9.919～25.564 ?g/g and 6.229～18.160 ?g/g in Hedyotis diffusa Willd from different sources. The linear range of caffeic acid was 0.005 0～0.200 0 ?g(r=0.999 9),the average recovery was 102.35%,RSD was 2.31%(n=6);The linear range of quercetin was 0.006 2～0.244 0 ?g(r=0.999 9),the average recovery was 101.84%,RSD was 1.79%(n=6);The linear range of campherenol was 0.007 8～ 0.310 6 ?g(r=0.999 9),the average recovery was 99.04%,RSD was 2.90%(n=6). CONCLUSION:The method for quantifying of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd is accurate and reliable.

Objective Matrix metalloproteinase-10 (MMP-10) has been shown to be highly associated with atherosclerosis.Recent studies showed that levels of MMP-10 were elevated in infarcted tissues in acute ischemic stoke.However,serum levels of MMP-10 in patients with acute ischemic stroke have never been studied previously.This study aims to investigate the serum levels of MMP-10 in patients with acute ischemic stroke,and evaluate the association of serum levels of MMP-10 with stroke subtypes based on Trial of Org 10 172 in acute stroke treatment classifications,the severity of stroke,risk factors and carotid artery plaque.Methods The circulating levels of MMP-10 were measured by enzyme linked immunosorbent assay in 194 subjects,including 109 patients who were diagnosed as acute ischemic stroke in the Department of Neurology,Shengjing Hospital,China Medical University from April to December 2012,and the 85 healthy controls.Results Patients with acute ischemic stroke had higher serum levels of MMP-10 compared with healthy controls (6.59 (6.07,7.31) μg/L vs 5.16 (3.87,5.94) μg/L,Z =8.33,P ＜ 0.01).NIHSS score had positive correlation with serum levels of MMP-10 (r =0.204,P =0.037).Classified by risk factors,we compared the MMP-10 levels of subsets,and results displayed that statistically significant difference existed between dyslipidemia subset and non-dyslipidemia subset (Z =2.07,P =0.042).MMP-10 levels had positive correlation with serum levels of LDL-cholesterol (r =0.248,P =0.040),but negative correlation with thrombin-activatable fibrinolysis inhibitor (TAFI;r =-0.208,P =0.030).The subset with unstable plaques had higher MMP-10 levels than that with stable plaque (6.62 (6.13,7.36) μg/L) vs 6.10 (6.00,6.46) μg/L,Z =2.12,P =0.034),implying the relationship of MMP-10 and atherosclerosis.Conclusions Patients with acute ischemic stroke have higher serum levels of MMP-10 compared with the healthy controls,and MMP-10 levels have positive correlation with the severity of stroke.MMP-10 is associated with the subtypes of stroke classified by risk factors,and dyslipidemia subset has higher levels of MMP-10 than that of non-dyslipidemia subset.MMP-10 has positive correlation with LDL-cholesterol,but negative correlation with TAFI.MMP-10 may be involved in the process of formation and disruption of unstable plaques,which contribute to the stenosis of arteries and onset of acute ischemic stroke.

Objective To explore the expression of c-Fos protein and character of mossy fiber sprouting(MFS) in hippocampus of rat with febrile seizures(FS).Methods Thirty-six 21-day-old male Sprague-Dawley rats were randomly divided into FS group,febrile control(FG) group and normal control(NG) group.FS was established by hyperthermal bath.Immune histochemistry and(Timm′s) staining were used to examine the expression of c-Fos protein in CA1 region and MFS in CA3 region of hippocampus.Results Excessive expression of c-Fos protein presented in the hippocampal CA1 region of FS group.The surface area percentage of c-Fos protein of FS group[(2.26?0.23)%] was higher than that of FG group[(1.08?0.19)%] and NG group[(0.71?0.14)%],there were significant difference between FS group and the other two groups(?~2=10.48 P

OBJECTIVETo observe the influences of quercetin (Que) on the contraction of small intestine smooth muscle of rabbits in vitro and explore the mechanism.

METHODSWith the isothermal perfusion of small intestine in vitro. The influences of quercetin on the spontaneous contraction of small intestine and contraction induced by Ach, histamine and Bacl2 were observed and the mechanism of quercetin was studied.

RESULTSQuercetin reduced the tension of contraction of small intestine smooth muscle in rabbits in a dose-depended manner. Quercetin could completely block the contraction of Bay K8644. Heparin could also block the inhibition of quercetin on small intestine smooth muscle but ruthenium red (RR) had no effect on the relaxation of quercetin. Nitro-L-arginine methylester(L-NAME) inhibited the relaxation of quercetin.

CONCLUSIONQuercetin inhibits the contraction of small intestine smooth muscle of rabbits in vitro. The mechanism may be related to increase NO concentration in small intestine smooth muscle so that it inhibits extracellular Ca2+ inflowing via cell membrane. And quercetin has effect on intracellular Ca2+ releasing via IP3 of sarcoplasmic reticulum.

Animals ; Calcium ; metabolism ; In Vitro Techniques ; Intestine, Small ; drug effects ; Muscle, Smooth ; drug effects ; physiology ; Quercetin ; pharmacology ; Rabbits ; Sarcoplasmic Reticulum ; drug effects ; metabolism

OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

Adult ; Aged ; Aged, 80 and over ; Benign Paroxysmal Positional Vertigo ; diagnosis ; drug therapy ; economics ; Female ; Humans ; Male ; Middle Aged ; Surveys and Questionnaires ; Treatment Outcome

OBJECTIVETo study the prevalence of iron deficiency in children between 6 months and 7 years and to study the diagnostic value of soluble transferrin receptor (sTfR) for iron deficiency in the children.

METHODSA total of 502 healthy children between 6 months and 7 years from Hangzhou City of Zhejiang Province were enrolled. Serum sTfR, serum ferritin (SF), serum iron (SI), total iron blinding capacity (TIBC), zinc protoporphyrin (ZPP), Hb, MCV and CRP levels were measured.

RESULTSThe prevalence rate of iron deficiency was 19.5% in children at ages of 6 months to 7 years. The prevalence rate of iron deficiency was the highest in infants (≤1 year old; 34.7%), followed by in toddlers (1-3 years old; 19.4%) and preschoolers (3-7 years old; 14.0%). The mean serum sTfR level in infants (2.02±0.73 mg/L) was significantly higher than that in toddlers (1.68±0.40 mg/L) and preschoolers (1.67±0.29 mg/L) (P<0.05).The best cut-off value of serum sTfR for the diagnosis of iron deficiency was 2.02 mg/L in infants (sensitivity: 70.3%, specificity: 82.2%). The best cut-off value was 1.85 mg/L in toddlers (sensitivity: 71.7%; specificity: 86.4%), and that was 1.85 mg/L in preschoolers (sensitivity: 77.8%; specificity: 88.6%). Serum sTfR was correlated with SF (r=0.107, P<0.05), TIBC (r=0.276, P<0.01), TS (r=-0.139, P<0.05), ZPP (r=0.175, P<0.01) and MCV (r=-0.140, P<0.01).

CONCLUSIONSIron deficiency is more prevalent in infants ≤1 year old. The mean serum level and the cut-off value of sTfR in infants are higher than in toddlers and preschoolers. Serum sTfR is an effective index for the diagnosis of iron deficiency in children, especially in infants≤ 1 year old.

Age Factors ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Iron ; deficiency ; Male ; Prevalence ; Receptors, Transferrin ; blood

OBJECTIVETo explore the different effects of rectal application of Neiyi Kangfu Suppository (NYKFS) on the expressions of cytochrome C (Cyt C) and Survivin in ectopic and eutopic endometrium in endometriosis (EMT) model rats.

METHODSEMT rats were randomly divided into 6 groups, the high- and low-dose NYKFS groups, the Danazol Capsule (DN) group, the Dan'e Fukang Soft Extract (DFE) group, the model group and the blank group. The expressions of Cyt C and Survivin were measured using immunohistochemical SP staining method.

RESULTS(1) After treatment, the IOD value of Cyt C in ectopic endometrium significantly increased in the high-dose and low-dose NYKFS groups respectively to 6.08 +/- 0.35 and 6.23 +/- 0.35, which was significantly higher than that in the model group (5.07 +/- 0.70) and DFE group (5.98 +/- 1.02) respectively (P < 0.05, P < 0.01); while those of Survivin in ectopic endometrium was significantly decreased to 5.73 +/- 0.93 and 5.62 +/- 0.93 and was significantly lower than that in the model group (6.01 +/- 1.16, P < 0.05 or P < 0.01). (2) The lowered eutopic endometrial level of Cyt C in EMT rats after treatment was significantly higher in the two NYKFS groups than that in the model group (P < 0.05); while that of eutopic endometrial Survivin was insignificantly different between the NYKFS groups and the model group or the blank group (P > 0.05).

CONCLUSIONNYKFS plays its role in inducing apoptosis by increasing the expression of Cyt C and decreasing the expression of Survivin in ectopic endometrium, but it up-regulates expression of Cyt C, has no obvious effect on Survivin expression in eutopic endometrium.

Animals ; Cytochromes c ; biosynthesis ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Endometriosis ; drug therapy ; metabolism ; pathology ; Endometrium ; drug effects ; metabolism ; pathology ; Female ; Immunohistochemistry ; Microtubule-Associated Proteins ; biosynthesis ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Suppositories

Adult ; Bile Ducts, Intrahepatic ; diagnostic imaging ; pathology ; Biopsy, Needle ; Caroli Disease ; complications ; diagnosis ; pathology ; Child ; Diagnosis, Differential ; Female ; Humans ; Liver ; diagnostic imaging ; pathology ; Liver Cirrhosis ; complications ; congenital ; diagnosis ; pathology ; Male ; Spleen ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed