This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.
Animals ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; metabolism ; pharmacology ; Mass Spectrometry ; Osteoporosis ; drug therapy ; Zebrafish

Acute Disease ; Adult ; Arsenic Poisoning ; pathology ; Humans ; Male ; Respiratory System ; pathology ; Young Adult

In this paper, the action of suet oil in the preparation of self-assembled micelles of the active flavonoids in Epimedium in the simulated human environment was researched. Twelve suet oil samples were collected from different growing areas and different positions of sheep or goat to simulate the formation of micelles. Then the effects of the fatty acids in suet oil on the preparation of self-assembled micelles were studied furthermore. The results showed that the micelles had a dispersed state and spherical smooth surface. To compare the diameter, potential, encapsulation efficiency and drug loading of the 12 batches micelles, the micelles prepared by the suet oil from Qinghai were more stable and had a higher encapsulation efficiency. The fatty acids in suet oil could promote the formation of self-assembled micelles, but the whole suet oil had a better effect. Above all the study, we confirmed that the suet oil promoted the formation of self-assembled micelles of the flavonoids in Epimedium, it laid foundation for further research about increasing the efficacy of Epimedium and improved the absorption of the active flavonoids in Epimedium.
Animals ; Chemistry, Pharmaceutical ; methods ; China ; Drug Carriers ; chemistry ; Electric Conductivity ; Epimedium ; chemistry ; growth & development ; Fatty Acids ; chemistry ; Flavonoids ; chemistry ; Geography ; Goats ; Humans ; Micelles ; Oils ; chemistry ; Sheep

To prepare sagittatoside B with epimedin B Hydrolyzed from cellulase. With the conversion ratio as the index, the effects of pH value, temperature, reaction time, dosage of enzyme and concentration of substrates on the conversion ratio were detected. L9 (3(4)) orthogonal design was adopted to optimize the preparation process. Hydrolyzed products were identified by MS, 1H-NMR, and 13C-NMR. The results showed that the optimum reaction conditions for the enzymatic hydrolysis were that the temperature was 50 degrees C, the reaction medium was pH 5.6 acetic acid-sodium acetate buffer solution, the concentration of substrates was 20 g x L(-1), the mass ratio between enzyme and substrate was 3: 5, and the relative molecular mass of the reaction product was 646.23. NMR data proved that the product was sagittatoside B. The process is simple and reliable under mild reaction conditions, thus suitable for industrial production.
Cellulase ; metabolism ; Drug Compounding ; methods ; Flavonoids ; chemistry ; Hydrogen-Ion Concentration ; Hydrolysis ; Temperature ; Time Factors

Heuristic teaching is propitious to cultivating students' ideation. We have used different heuristic modes for example problems, stories, contrast, associationin and so on to cultivate medical students' ideation in pathobiology and immunology teaching and acquired good effect.

OBJECTIVEObserve the clinical effect of removable lingual arch plus auxiliary spring for convenient dental (especially mandibular) arch expansion.

METHODSSeventeen patients with dental arch constriction complicated by mild dental crowding were enrolled in this study, including 9 requiring maxillary arch expansion and 8 necessitating mandibular expansion. The patients were divided to two groups at random for arch expansion with removable lingual arch plus auxiliary spring and with Quad-helix, respectively, and the effect of arch expansion was compared between the two groups.

RESULTSAfter 8 weeks of arch expansion, the average width of U4-U4 and U5-U5 in removable lingual arch group was enlarged by 2.2 mm and 3.0 mm, and that in Quad-helix group by 2.3 mm and 3.5 mm, respectively, showing no significant differences between the two groups (P>0.05). After 12 weeks of treatment, the average width of L4-L4 and L5-L5 in the former group was enlarged by 2.3 mm and 2.5 mm, respectively, significantly greater than that in the latter group (1.0 mm and 1.2 mm, P<0.05).

CONCLUSIONRemovable lingual arch plus auxiliary spring allows significant expansion of the middle segment of the dental arch (bicuspids), and can be more effective than Quad-helix for mandibular arch expansion.

Adolescent ; Child ; Dental Arch ; pathology ; Humans ; Malocclusion ; therapy ; Orthodontic Appliances, Removable ; Orthodontics, Corrective ; methods ; Palatal Expansion Technique ; instrumentation ; Young Adult

Objective To observe the influence of rotenone on the distribution of alpha-synuclein (ASN) in rat model of Parkinson's disease (PD). Methods Wistar rats were randomly divided into two groups and received 2 mg/kg rotenone (s.c.) or sunflower oil (as control group) for about 4 weeks. The hippocampus, substantia nigra and striatum of brain were observed. Hematoxylin and eosin stain were used to observe the Lewy body like inclusion. The expression of tyrosine hydroxylase (TH) or ASN protein was determined by anti-TH or anti-alpha-synuclein immunohistochemistry, respectively. Results In control rats, ASN protein distributed widely in brain, especially in hippocampus, cortex and striatum. Rotenone obviously increased TH positive neurons and fibers loss in substantia nigra and striatum (P < 0.05). In rotenone treated rats, ASN positive cells increased in global brain but not distributed in an even manner. In substantia nigra, ASN positive stuff was found aggregate in both cytoplasm and nucleus, and some formed spherical inclusion; in striatum, ASN positive neurites end aggregated and agglomerated around neurons; and in hippocampus, few dot-like ASN were aggregated in cell body, and no notable change was found in nucleus. Conclusion In rotenone administrated PD rats, ASN protein aggregated in several brain regions but most obviously in striatum and substantia nigra, and the distribution region of ASN was changed from peri-synapse to the cytoplasm and nucleus of dopaminergic neuron.

OBJECTIVETo investigate the epidemiological characteristics of human papillomavirus (HPV) infection in men attending a sexually transmitted diseases (STD) clinic in Hangzhou area.

METHODSMale subjects (n=375) aged 18-70 years, attending the STD clinic were recruited. Urethral swabs were assessed for HPV DNA using polymerase chain reaction (PCR) with the consensus primers MY09/11. HPV genotypes of positive PCR products were determined by restriction fragment length polymorphisms and direct sequence analysis.

RESULTSOf the 375 swabs collected, 305 (81.3%) yielded sufficient DNA for the subsequent HPV analysis. Among the 305 subjects, the prevalence of HPV was 13.8%. Nononcogenic HPV types were found in 8.5% (26/305) of subjects, oncogenic types in 4.3% (13/305), and multiple types in 1.0% (3/305). The prevalence of HPV infection was higher in subjects from urban area than in those from rural area (P < 0.05). The prevalence was also higher in those who received fewer years of education (P < 0.05) and those who had more sex partners (P < 0.05).

CONCLUSIONSHPV infection among men at high risk is not uncommon. The detection rate of HPV DNA is significantly related to some sociodemographic factors, such as residence, educational level and the number of sex partners.

Adolescent ; Adult ; Aged ; Ambulatory Care Facilities ; China ; epidemiology ; Humans ; Male ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; epidemiology ; virology ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sexually Transmitted Diseases ; prevention & control

OBJECTIVETo prepare CIT microemulsion and to investigate its properities and the absorption character in rat intestine in situ.

METHODThe optimum formulation of the blank microemulsion was selected by pseudo tertiary phase diagrams and the CIT microemulsion was prepared based on the blank microemulsion. The appearance, particle diameter distribution and Zeta potential were investigated by electron microscopy and grainsize analyzer. The entrapment efficiency was determined by sephadex column chromatography. An in situ single pass intestine perfusion method was used to investigate the intestinal absorption of CIT microemulsion. HPLC method for determination of CIT in the intestinal flux was established.

RESULTThe formulation was OP-glycerol-water-ethylis oleas (3:3:4:2). CIT microemulsion in electron microscopy was consisted of small spherical drop. The mean diameter was 61.7 nm, Zeta potential was -46.3 mV. The Entrapment efficiency was (92.1 +/- 1.74)% (n=3). Both CIT and CIT microemulsion had been absorbed in general intestinal tract,and the absorption of CIT microemulsion was significantly high compared with CIT in duodenum, colon, jejunum and ileum (P < 0.01).

CONCLUSIONThe preparation technology of CIT microemulsion was feasible,and the microemulsion system might improve the absorption of CIT in the intestinal tract.

Animals ; Drug Compounding ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Emulsions ; chemistry ; Intestinal Absorption ; Models, Animal ; Particle Size ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo screen novel miRNAs targeting EZH2 3' untranslated region (UTR) in recombinational MCF-7 breast cancer cells over-expressing EZH2 3' UTR and quantitative analyze the expressions of the screened miRNA in breast cancer cells and tissues.

METHODSA lentiviral library was transfected into the recombinant cell line MCF-7. The cells were screened with cytotoxic agents before extraction of the genome for amplification of the miRNA precursors using PCR. The screened miRNAs were identified with sequence analysis and their expressions were analyzed quantitatively with real-time PCR in breast cancer cells and tissues.

RESULTSSeven miRNAs were screened from the recombinant MCF-7 cells, namely miR-15b, miR-16-2, miR-181b2, miR-217, miR-224, miR-329-1, and miR-487b, all of which failed to be predicted by bioinformatics software. Real-time PCR showed that miR-217, miR-329-1, and miR-487b were over-expressed in MCF-7 cells, and the expression of miR-15b and miR-16-2 was significantly increased in cancer tissues compared with the adjacent tissues (P<0.05).

CONCLUSIONNovel targeted miRNAs that can not be predicted by bioinformatics software were successfully screened from MCF-7 breast cancer cells over-expressing EZH2 3' UTR. These miRNAs are expressed differentially between normal breast cells and breast cancer tissues.

3' Untranslated Regions ; Breast Neoplasms ; genetics ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Female ; Gene Expression Profiling ; Humans ; Lentivirus ; genetics ; MicroRNAs ; genetics ; Polycomb Repressive Complex 2 ; genetics