The present study is to establish Caco-2/HT29-MTX co-cultured cells and investigate the transport capability of PLGA nanoparticles with different surface chemical properties across Caco-2/HT29-MTX co-cultured cells. PLGA-NPs, mPEG-PLGA-NPs and chitosan coated PLGA-NPs were prepared by nanoprecipitation method using poly(lactic-co-glycolic acid) as carrier material with surface modified by methoxy poly(ethylene glycol) and chitosan. The particle size and zeta potential of nanoparticles were measured by dynamic light scattering. Coumarin 6 was used as a fluorescent marker in the transport of nanoparticles investigated by confocal laser scanning microscopy. The transport of furanodiene (FDE) loaded nanoparticles was quantitively determined by high performance liquid chromatography. Colchicine and nocodazole were used in the transport study to explore the involved endocytosis mechanisms of nanoparticles. Distribution of the tight junction proteins ZO-1 was also analyzed by immunofluorescence staining. The results showed that the nanoparticles dispersed uniformly. The zeta potential of PLGA-NPs was negative, the mPEG-PLGA-NPs was close to neutral and the CS-PLGA-NPs was positive. The entrapment efficiency of FDE in all nanoparticles was higher than 75%. The transport capability of mPEG-PLGA-NPs across Caco-2/HT29-MTX co-cultured cells was higher than that of PLGA-NPs and CS-PLGA-NPs. Colchicine and nocodazole could significantly decrease the transport amount of nanoparticles. mPEG-PLGA-NPs could obviously reduce the distribution of ZO-1 protein than PLGA-NPs and CS-PLGA-NPs. The transport mechanism of PLGA-NPs and mPEG-PLGA-NPs were indicated to be a combination of endocytosis and paracellular way, while CS-PLGA-NPs mainly relied on the endocytosis way. PEG coating could shield the surface charge and enhance the hydrophilicity of PLGA nanoparticles, which leads mPEG-PLGA-NPs to possess higher anti-adhesion activity. As a result, mPEG-PLGA-NPs could penetrate the mucus layer rapidly and transport across Caco-2/HT29-MTX co-cultured cells.
Biological Transport ; Caco-2 Cells ; Chitosan ; chemistry ; Coated Materials, Biocompatible ; chemistry ; Coculture Techniques ; Drug Carriers ; Furans ; administration & dosage ; chemistry ; metabolism ; HT29 Cells ; Heterocyclic Compounds, 2-Ring ; administration & dosage ; chemistry ; metabolism ; Humans ; Lactic Acid ; chemistry ; Nanoparticles ; Particle Size ; Polyethylene Glycols ; chemistry ; Polyglycolic Acid ; chemistry ; Zonula Occludens-1 Protein ; metabolism

Objective To study the curative effect and adverse reaction of Oxcarbazepine(OXC)on treating epilepsy in children.Me-thods One hundred and twenty-nine children with different types of epilepsy were orally given OXC,and the drug dose was added gradually.According to the seizure frequency,the cases were divided into 2 groups.Group A:more than 3 seizures occurred in 3 months prior to to take OXC;group B:more than 3 seizures occurred in 1 year prior to taking OXC.After 5 months and 1 year from beginning to take OXC,the original curative effect of the 2 groups was evaluated,respectively;on the other hand,the adverse reaction and the retention were studied.Results 1.Original effect:the rate of seizure-free was 45.8% and the total curative efficiency was 66.7% in group A(n=47);the rate of seizure-free was 92.3% in group B(n=13);in 60 partial epilepsy children,the rate of seizure-free was 56.7% and the total curative efficiency was 73.3%;Both rates were 62.2% and 75.6% of patients with OXC monotherapy and 40%,66.7% of patients were given OXC in combination with another antiepileptic drugs.2.Drug adverse reaction:24% of patients were found to have adverse reaction and most of the symptoms were light and most transient.3.Tolerability:patients' retention of OXC in 1 year was 72.2%,and in 2 years was 80.0%.Conclusions The antiepileptic effect of OXC is satisfactory and adverse reaction is light and mostly transient,OXC as a new antiepileptic drug is well tolerated as monotherapy and adjunctive therapy and should be used extensively.J Appl Clin Pediatr,2009,24(1):53-55

Alanine Transaminase ; metabolism ; Animals ; Liver ; drug effects ; Male ; Mice ; Plant Extracts ; pharmacology

Objective Color doppler ultrasonography of chronic Keshan disease (CKD) was evaluated to provide evidences for clinic diagnosis of the disease. Methods From September to Novermber 2009, according to "Diagnostic criteria of Keshan disease" (GB 17021-1997), 64 cases of CKD were randomly sampled from five Keshan diseased districts in Shandong province, Zoucheng, Sishui, Yishui, Wulian, Jvxian, and Pingyi as patient group. Thirty four healthy volunteers being checked up by Shandong Institute for Endemic Diseases Control and Research were put in control group. All the subjects were examined with Color doppler ultrasonography. The indexes of cardiac structure, left ventricular (LV) systolic function and LV diastolic function were measured.Results Left atrial internal diameter, LV end-diastolic internal diameter, LV end-systolic internal diameter, right ventricular diameter, aorta diameter, right atrial transverse diameter, right atrial long diameter and left ventricle mass of the patient group[(35.38 ± 6.89), (61.57 ± 8.61), (45.39 ± 10.29), (17.22 ± 3.79), (28.69 ± 2.81),(38.00 ± 6.05), (42.68 ± 8.65)mm, (283.22 ± 103.12)g] were higher than that of control group[(26.70 ± 3.27),(45.41 ± 4.93), (26.91 ± 4.35), (13.76 ± 2.27), (24.09 ± 2.89), (31.50 ± 3.32), (35.82 ± 3.14) mm, (156.03 ±39.86)g, t = 6.93, 10.09, 9.98, 4.87, 7.64, 5.81, 4.46, 6.90, all P＜ 0.05]. The LV ejection fraction and fractional shortening of the left ventricular of the patient group[(49.25 ± 14.33)%, (26.11 ± 9.17)%] were lower than that of control group[(73.88 ± 4.04)%, (42.88 ± 3.62)%, t = - 9.79, - 10.22, all P＜ 0.05]. Diffuse hypokinetic motion of the left ventricle reduced in 95% (61/64) of CKD patients, and 5% (3/64) of CKD patients had segmental LV dyskinesia. Seventy five per sent(48/64) of the patients accompanied with mitral regurgitation, and 39% (26/64) of these cases accompanied with tricuspid regurgitation. Meaningful Mitral or tricuspid regurgitation was not found out in control group. Conclusions The CKD patients' bore of atrio-ventricular cavity and LV mass are enlarged, and their motion of ventricle is reduced or partly reduced. They have poor heart function. Mitral regurgitation are more than tricuspid regurgitation. Color doppler Ultrasonography is important in diagnosis of chronic Keshan discase.

OBJECTIVETo establish osteoblast model, primary cilla model was removed by chloral hyrate, observe effects of osteoblast primary cilla moved on enhancing ALP staining and calcified nodules staining in electromagnetic field.

METHODSThree 3-day-old male SD rats weighed between 6 and 9 g were killed, cranial osteoblast was drawed and adherencing cultured respectively. Cells were subcultured and randomly divided into 4 groups until reach to fusion states. The four groups included chloral hydrate non-involved group (control group), 2 mM, 4 mM and 8 mM chloral hydrate group, and cultured in 37 °C, 5% CO2 incubator for 72 h. Morphology of primary cilla was observed by laser confocal scanning microscope, and incidence of osteoblast primary cilia was analyzed by Image-Pro Plus 6.0 software. Cells in the correct concentration group which can removed cillia most effectively were selected and divided into 3 groups, including control group (C), Electromagnetic fields group (EMFs), and EMFs with 4 mM chloral hydrate group. DMEM nutrient solution contained 10%FBS were added into three groups and cultured for 9 days and formation of ALP were observed by histochemical staining of alkaline phosphatase. After 12 days' cultivation, formation of mineralization nodes was observed by alizarin red staining.

RESULTSCompared with control group and 2mM chloral hydrate group,4 mM chloral hydrate group could effectively remove osteoblast primary cilla (P<0.01). Removal of osteoblast primary cilla could weaken the formation of ALP and mineralization nodes in osteoblast in EMFS. Compared with EMFs group, the area of ALP and mineralization nodes in EMFs with 4 mM chloral hydrate group were decreased obviously (P<0.01).

CONCLUSION4mM chloral hydrate could effectively remove osteoblast primary cilia. Primary cilla participate in EMFs promoting formation of ALP and mineralization nodes in osteoblast and provide new ideas for exploring mechanism of EMFs promoting osteoblast maturation and mineralization.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Culture Techniques ; instrumentation ; methods ; Cells, Cultured ; Chloral Hydrate ; pharmacology ; Cilia ; drug effects ; enzymology ; physiology ; Male ; Osteoblasts ; cytology ; enzymology ; Rats ; Rats, Sprague-Dawley

Eight compounds were isolated from the stems of Brucea mollis by various chromatographic techniques such as column chromatography on silica gel and Sephadex LH-20, and preparative HPLC, and their structures were elucidated as bruceolline O (1), 1-(1-beta-glucopyranosyl)-1H-indole-3-carbaldehyde (2), canthin-6-one (3), 11-hydroxycanthin-6-one (4), 9-methoxycanthin-6-one (5), 4-methoxycanthin-6-one (6), infractin (7), and beta-carboline-1-propionic acid (8). The cytotoxic activities of compounds 1-8 against HCT-8 and A549 human cell lines were determined, but none of them exhibited significant activity (IC 50 > 10 micromol x L(-1)). Among them, compound 1 is a new indole alkaloid, and compounds 2 and 5-7 were isolated from this plant for the first time.
Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Brucea ; chemistry ; Carbolines ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Humans ; Indole Alkaloids ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

An analysis method has been established to test 27 elements (Li, Be, B, Mg, Al, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Sr, Mo, Cd, Sn, Sb, Ba, La, Hg, Pb, Bi) in Maca nationality's medicine with microwave digestion-ICP-MS. Sample solutions were analyzed by ICP-MS after microwave digestion, and the contents of elements were calculated according to their calibration curves, and internal standard method was adopted to reduce matrix effect and other interference effects. The experimental results showed that the linear relations of all the elements were very good; the correlation coefficient (r) was 0.9994-1.0000 (Hg was 0.9982) ; the limits of detection were 0.003-2.662 microg x L(-1); the relative standard deviations for all elements of reproducibility were lower than 5% (except the individual elements); the recovery rate were 78.5%-123.7% with RSD lower than 5% ( except the individual elements). The analytical results of standard material showed acceptable agreement with the certified values. This method was applicable to determinate the contents of multi-elements in Maca which had a high sensitivity, good specificity and good repeatability, and provide basis for the quality control of Maca.
Lepidium ; chemistry ; Mass Spectrometry ; methods ; Microwaves ; Reproducibility of Results ; Trace Elements ; chemistry ; isolation & purification

OBJECTIVETo observe therapeutic effect of acupoint embedding thread therapy on ulcerative colitis and to study the mechanism.

METHODSThe rat model of ulcerative colitis was developed with 2,4-dinitrochlorobenzene method. The 20th day after the end of model development, they were randomly divided into a model group, a salicylazosulfapyridine (SASP) treatment group and an acupoint embedding thread therapy group. The SASP treatment group were treated with intragastrical perfusion of 4.5% SASP suspension, and the acupoint embedding thread therapy group were treated with embedding thread at "Zusanli" (ST 36) and "Shangjuxu" (ST 37). Anti-neutrophil cytoplasmic antibody (ANCA) was detected with indirect immunofluorescence, and the pathological changes were investigated with pathological tissue section technique.

RESULTSThe positive rate of ANCA reached to 62.5% in the rat with ulcerative colitis with significant differences among the 3 groups. After treatment, the positive rate of ANCA was decreased in the two treatment groups (P < 0.05, P < 0.01).

CONCLUSIONAcupoint embedding thread therapy has definite therapeutic effect on ulcerative colitis and the mechanism is possibly related with the decrease of ANCA.

Acupuncture Points ; Animals ; Antibodies, Antineutrophil Cytoplasmic ; Colitis, Ulcerative ; therapy ; Rats

To investigate the effects of particle size, mPEG molecular weight, coating density and zeta potential of monomethoxyl poly(ethylene glycol)-poly(lactic-co-glycolic acid) (mPEG-PLGA) nanoparticles on their transportation across the rat nasal mucosa, mPEG-PLGA-NPs with different mPEG molecular weights (M(r) 1 000, 2 000) and coating density (0, 5%, 10%, 15%) and chitosan coated PLGA-NP, which loaded coumarin-6 as fluorescent marker, were prepared with the nanoprecipitation method and emulsion-solvent evaporation method, and determine their particle size, zeta potential, the efficiency of fluorescent labeling, in vitro leakage rate and the stability with the lysozyme were determined. The effects of physical and chemical properties on the transmucosal transport of the fluorescent nanoparticles were investigated by confocal laser scanning microscopy (CLSM). The result showed that the size of nanoparticles prepared with nanoprecipitation method varied between 120 and 200 nm; the size of nanoparticles prepared with emulsion-solvent evaporation method varied between 420 and 450 nm. Nanoparticles dispersed uniformly; the zeta potential of PLGA-NPs was negative; mPEG-PLGA-NPs was close to neutral; chitosan coated PLGA-NPs was positive; and the efficiency of fluorescent labeling were higher than 80%. In vitro leak was less than 5% within 4 h and nanoparticles were basically stable with lysozyme. The CLSM results show that the transportation efficiency of mPEG-PLGA-NPs with a high PEG coating density and high mPEG molecular weight was significantly higher than that of uncoated PLGA nanoparticles and also that of chitosan coated PLGA-NPs (P < 0.05). The hydrophilcity, zeta potential and particle size of nanoparticles play important roles on the efficiency of mPEG-PLGA nanoparticles to transport across the rat nasal mucosa.
Animals ; Biological Transport ; Chitosan ; chemistry ; Drug Carriers ; chemistry ; Female ; Male ; Microscopy, Confocal ; Molecular Weight ; Nanoparticles ; Nasal Mucosa ; metabolism ; Particle Size ; Polyesters ; chemistry ; pharmacokinetics ; Polyethylene Glycols ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

Coronary Vessel Anomalies ; diagnosis ; Electrocardiography ; Humans ; Infant ; Pulmonary Artery ; abnormalities