OBJECTIVEAn accurate and reliable analytical method for-simultaneous determination of six active components (scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C) in plants of Erycibe was developed.

METHODScopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in the samples were well separated in analytical HPLC by gradual elution with methanol-0.1% formic acid solution. The chromatographic condictions: Agilent Poroshell 120 EC-C18 column, flowing rate being 1 mL x min(-1), detecting wavelength at 345 nm.

RESULTGood linearities of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C were in the range of 0.026 8-2.68, 0.027 0-2.70, 0.008 1-0.81, 0.018 8-1.88, 0.017 6-1.76, 0.019 6-1.96 μg, respectively (r > 0.999 6). The average recoveries of the six components were 98.1%, 98.7%, 100.8%, 100.4%, 99.7%, 101.1%; the relative standard deviations were 2.67%, 2.86%, 2.62%, 1.98%, 2.76%, 2.19%.

CONCLUSIONThe method is simple, feasible and reproducible and can be used for the quality control of plants of Erycibe.

China ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Convolvulaceae ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Scopoletin ; analysis

OBJECTIVETo isolate and identify Streptococcus mutans (Sm) and Streptococcus sobrinus (Ss) in dental plaque of children with high dmft and no caries by selective medium, biochemical methods and arbitrarily primed-polymerase chain reaction (AP-PCR).

METHODSA total of 401 3-4-year-old children from seven kindergartens were recruited using cluster sampling and their dental caries status were examined. From 30% of children with the highest dmft score (dmft >/= 5), 20 children were chosen randomly as test group and 20 age and gender-matched caries-free children were selected as control. Plaque samples were collected from buccal surfaces of the molars and plated onto TYCSB plate. Sm and Ss were primarily identified by colony morphology and biochemical characteristics. Then chromosomal DNA of the strains was isolated and Sm or Ss were confirmed by AP-PCR.

RESULTSThe proportion positive for Sm and Ss in children with high dmft was 100% and 40% respectively while that in caries-free children was 75% and 5% by AP-PCR analysis. The differences were statistically significant between the two groups.

CONCLUSIONSThe proportions positive for Sm and Ss detected by AP-PCR method were significantly higher in children with high dmft than in caries-free children and it is a risk factor for high dmft in deciduous teeth harboring Sm and Ss.

Child, Preschool ; Dental Caries ; microbiology ; Dental Plaque ; microbiology ; Female ; Humans ; Male ; Streptococcus mutans ; genetics ; isolation & purification ; Streptococcus sobrinus ; genetics ; isolation & purification

Electroencephalography ; Female ; Humans ; Hyperbilirubinemia, Neonatal ; physiopathology ; Hypoxia-Ischemia, Brain ; physiopathology ; Infant, Newborn ; Infant, Newborn, Diseases ; physiopathology ; Male ; Retrospective Studies

Objective　To observe the changes of the microvascular permeability after blunt chest trauma (BCT), endotoxemia and their combined injury in rats. Methods　After the establishment of the rat models of BCT, endotoxemia and their combined injury in the right lungs, the fluorescein sodium (FINa) content was measured with flurospectrophotometer in lungs 0.5, 1, 4 and 8 h after injury. Results　There was an early obvious increase of the microvascular permeability in the impact lateral (peak at half an hour after injury), and a delayed increase in the contralateral lung (peak at the 8th h) in the BCT group. The FINa content was higher in endotoxemia group than in the BCT group(P<0.05), and lower than that in the combined injury group(P<0.05) in the contralateral lung. Conclusion　Results indicate that there were different pathophysiologic processes among the 3 kinds of injury and the FINa content is a useful index to manifest the changes of microvascular permeability in tissues.

Objective To evaluate the enhanced magnification endoscopy in the diagnosis of Barrett esophagus,and to explore the relationship between mucosal surface patterns and pathological epithelial types of Barrett esophagus.Methods Enhanced magnification endoscopy was performed 'after spraying 2%-3% acetic acid on the surface of distal esophagus in 40 Barrett esophagus patients.Mucosal specimen were biop- syed.Results According to the mucosal types of Toyoda in 2003,there were three mucosal types:Ⅰ dot pat- tern 7(17.5%),5 of 7(71.4%)fundie type,Ⅱ reticular pattern 24(60.0%),16 of 24(66.7%)fundic type,Ⅲ cerebroid/villous 9(22.5%),intestinal metaplasia or dysplasia.Conclusion Enhanced magnifi- cation endoscopy helps to identify areas with intestinal metaplasia and dysplasia,and is useful in the diagno- sis of Barrett esophagus.

In order to explore the possibility of human adenovirus infection with tree shrews,the neutralizing antibody ti-ters of five kinds of human adenoviruses (HAdv)in the serum of tree shrews were analyzed.The levels of Ad3,Ad4,Ad7, Ad14 and Ad55 neutralizing antibody were detected by virus neutralization test.The results showed that the positive rate of four adenoviruses in group B were higher than Ad4 in group E,and the positive rates respectively were Ad14 (55.88%),Ad3 (47.06%),Ad55 (29.71%),Ad7 (14.71%)and Ad4 (8.82%).The antiserum mainly mixed with Ad3,Ad14 and Ad55 anti-body.Five species of human adenovirus can be naturally infected with tree shrews.Tree shrews are used as experimental ani-mals to establish human adenovirus infection model is alternative.

Animals ; Hydrogen Sulfide ; metabolism ; Hypertension, Portal ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley

PURPOSE: Although the interferon α (IFNα) signaling and the paired-like homeodomain transcription factor 2 (PITX2) have both been implicated in the progression of breast cancer (BCa), it remains obscure whether these two pathways act in a coordinated manner. We therefore aimed to elucidate the expression and function of PITX2 during the pathogenesis of endocrine resistance in BCa. MATERIALS AND METHODS: PITX2 expression was assessed in BCa tissues using quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunohistochemistry and in experimentally induced letrozole-resistant BCa cells using RT-qPCR and immunoblotting. Effects of PITX2 deregulation on BCa progression was determined by assessing MTT, apoptosis and xenograft model. Finally, using multiple assays, the transcriptional regulation of interferon-inducible transmembrane protein 1 (IFITM1) by PITX2 was studied at both molecular and functional levels. RESULTS: PITX2 expression was induced in letrozole-resistant BCa tissues and cells, and PITX2 induction by IFNα signaling powerfully protected BCa cells against letrozole insult and potentiated letrozole-resistance. Mechanistically, PITX2 enhanced IFNα-induced AKT activation by transactivating the transcription of IFITM1, thus rendering BCa cells unresponsive to letrozoleelicited cell death. Additionally, ablation of IFITM1 expression using siRNA substantially abolished IFNα-elicited AKT phosphorylation, even in the presence of PITX2 overexpression, thus sensitizing BCa cells to letrozole treatment. CONCLUSION: These results demonstrate that constitutive upregulation of PITX2/IFITM1 cascade is an intrinsic adaptive mechanism during the pathogenesis of letrozole-resistance, and modulation of PITX2/IFITM1 level using different genetic and pharmacological means would thus have a novel therapeutic potential against letrozole resistance in BCa.
Apoptosis ; Breast Neoplasms ; Breast ; Cell Death ; Heterografts ; Immunoblotting ; Immunohistochemistry ; Interferons ; Phosphorylation ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Small Interfering ; Transcription Factors ; Transcriptional Activation ; Up-Regulation

BACKGROUNDMany studies have suggested that the imbalance of angiogenic factor and anti-angiogenic factor expression contributes significantly to the development of choroidal neovascularization (CNV), and ultrasound microbubble combination system can increase the gene transfection efficiency successfully. This study was designed to investigate whether ultrasound-mediated microbubble destruction could effectively deliver therapeutic plasmid into the retina of rat, and whether gene transfer of pigment epithelium-derived factor (PEDF) could inhibit CNV.

METHODSHuman retinal pigment epithelial cells were isolated and treated either with ultrasound or plasmid alone, or with a combination of plasmid, ultrasound and microbubbles to approach feasibility of microbubble-enhanced ultrasound enhance PEDF gene expression; For in vivo animal studies, CNV was induced by argon lasgon laser in rats. These rats were randomly assigned to five groups and were treated by infusing microbubbles attached with the naked plasmid DNA of PEDF into the vitreous of rats followed by immediate ultrasound exposure (intravitreal injection); infusing liposomes with the naked plasmid DNA of PEDF into the vitreous (lipofectamine + PEDF); infusing microbubbles attached with PEDF into the orbit of rats with ultrasound irradiation immediately (retrobular injection); infusing microbubbles attached with PEDF into the femoral vein of rats with exposed to ultrasound immediately (vein injection). The CNV rats without any treatment served as control. Rats were sacrificed and eyes were enucleated at 7, 14, and 28 days after treatment. Gene and protein expression of PEDF was detected by quantitative real-time RT-PCR, Western blotting and immunofluorescence staining, respectively. The effect of PEDF gene transfer on CNV was examined by fluorescein fundus angiography.

RESULTSIn vitro cell experiments showed that microbubbles with ultrasound irradiation could significantly enhance PEDF delivery as compared with microbubbles or ultrasound alone. In the rat CNV model, transfection efficiency mediated by ultrasound/microbubbles was significantly higher than that by lipofectamine-mediated gene transfer at 28 days after treatment. The study also showed that with the administration of ultrasound-mediated microbubbles destruction, the CNV of rats was inhibited effectively.

CONCLUSIONSUltrasound-microbubble technique could increase PEDF gene transfer into rats' retina and chorioid, in association with a significant inhibition of the development of CNV, suggesting that this noninvasive gene transfer method may provide a useful tool for clinical gene therapy.

Animals ; Cells, Cultured ; Choroidal Neovascularization ; prevention & control ; Eye Proteins ; genetics ; Female ; Genetic Therapy ; Humans ; Microbubbles ; Nerve Growth Factors ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Long-Evans ; Retina ; metabolism ; Serpins ; genetics ; Transfection ; Ultrasonics