Advanced Materials Research Vols. 781-784

Abstract: Leaf extract of Plantago asiatica L. could inhibit the activity of cholinesterase slightly. Leaf extract of Iuicium verum and Reineckea carnea showed no effects on the activity of cholinesterase respectively. Leaf extract of Quercifilix zeylanica (Houtt.) Cop. could activate the activity of cholinesterase slightly. Leaf extract of Asparaguscochinchinensis(Lour.) Merr, Patrinia villosa Juss., Teucrium viscidum Bl., Aregelia spertabilis could activate the activity of cholinesterase evidently.Especially the leaf extract of Ajuga ciliata Bunge could activate the activity of cholinesterase markedly, with the activity being twice as the original one.Otherwise, stem extract of Euphorbia tirucalli could also activate the activity of cholinesterase evidently.

Abstract: Cerebrosides were extracted from Asterina pectinifera using ethanol extraction and silica gel column chromatography. The identification of the cerebrosides was established based on thin layer chromatography (TLC), UV, IR and HRMS. The cerebrosides exhibited moderate cytotoxic activities against Hela and BEL7402 cell lines. At the concentration range of 10-400μg/ml, cytotoxicity of cerebrosides on two human tumor cells showed a dose-and time-dependent manner.

Abstract: Ultrasonic-Assisted extraction (UAE) was applied for saponins extraction from Ophiopholis mirabilis and response surface methodology (RSM) was used to optimize the extraction technology. A first set of experiments were designed to optimize water bath temperature, ethanol concentration, the ratio of liquid to solid, ultrasonic time (min) and ultrasonic power (W) as the main variables affecting the extraction effect. A Box-Behnken design consisting of 15 experimental runs was then applied and a second-order polynomial model was used to explain the interactions between the main factors and the yield of saponins. The developed mathematical model was found to fit well with the experimental data and the optimized conditions were ultrasonic pretreatment time 30.9 min, ethanol concentration 64%, liquid /solid ratio 35:1, ultrasonic power 300W and water bath 70°C. Corresponding saponins content was 0.574%.

Abstract: A preparation of a bio-material of polymer beads used for immuno-assay is presented, in which the probe antibody is site-specific immobilized on the polystyrene microbead by the surface hydrazide activation and the oxidized glycochains linkage of antibody. Using adipic dihydrazide, free hydrazide-bearing matrix was acquired on carboxyl polystyrene microbead. An iron (III)-phenanthroline spectrophotometric method was developed to evaluate the activation efficiency. The dependence of probe antibody oxidization on the reaction time and the oxidant concentration was investigated. Also, an alternative strategy of antibody immobilization via traditional COOH/-NH₂ linkage was illustrated for a comparison with this site-specific immobilization in the aspects of the total antibody binding amount and the site-specific binding amount. The results showed that the site-specific efficacy using glycochains linkage-based binding was three times better than that using traditional amino-based binding, which indicated a higher sensitivity for application in immuno-assay.

Abstract: Mastitis is most critical disease in dairy cows and causes huge cost in the dairy industry. To prevent and treat it, it is important to understand the mechanisms of immune function in the mammary gland. Innate immunity is non-specific acute-response immune function. Some components of innate immunity in the mammary gland are found, e.g. Lingual Antimicrobial Peptide (LAP), Lactoferrin (LF). These components are found to be localized in the alveolar epithelium of mammary gland. LAP belongs to the β-defensin family, and plays a crucial role in killing a large variety of microorganisms. LF belongs to an iron-binding glycoprotein and has antibacterial activity. It is reported that LF has been localized immunohistochemically in mammary epithelial cells of lactating cows. Our previous study revealed that secretion of LAP into milk proceeded to that of LF after lipopolysaccharide (LPS) injection into the mammary gland. From this result, it is hypothesized that immunohistochemistry probably shows positive to either LF or LAP but not both in the alveolus vs epithelium in the mammary gland. Therefore, the aim of the present study is to investigate the immunolocalization of LAP and LF in the same bovine mammary tissue. Bovine mammary tissues were collected in the slaughterhouse and were fixed with neutralized formalin immediately. Paraffin sections (2-um thickness) were processed with antigen retrieval treatment followed by blocking with casein milk. Sections were cultured with LF antibody or LAP antibody. Immunoreaction products were visualized by incubation with a DAB. LAP and LF were localized in the cytoplasm of epithelial cell of alveolus. In some cases, LAP and LF were seen clearly in the same alveoli of section. In other cases, some epithelial cells were stained only LAP, but not LF, and other epithelial cells of alveolus were stained only LF, but not LAP. These results suggest the possibility that LAP and LF are differentially synthesized in the alveolar epithelium and may support our previous findings that their secretion occurs at the different time course.

Abstract: Mixed strains were used to increase the hydrolysis degree of materials during fermenting. Tilapia scraps and soybean meal were mixed by 1:1 and Bacillus subtilis, yeast and lactic acid bacteria were added. The optimal quantity of strains was determined by hydrolysis degree through single factor experiments and orthogonal experiments. After all, The molecular weight distribution of raw material and products was analyzed by high performance size exclusion chromatography (HPSEC). The results showed that the optimum conditions were as follows: the inoculation amount of Bacillus subtilis, the yeast and the lactic acid bacteria was 0.6‰, 0.06‰ and 0.0015‰, respectively. And the hydrolysis degree of materials increased from 10.3% to 27.3%. The hydrolysis degree of materials increased markedly by adding mixed strains.

Abstract: In this study, we aim to investigate the effect of curcumin on the expression of MAP2 in hippocampus in APP/PS1 double transgenic mice. AD model was established with APP/PS1 double transgenic mice, which were fed for 6 months with different concentration of curcumin diet. Immunohistochemistry were applied to evaluate the expressive of MAP2 in hippocampus of transgenic mice. The expression of MAP2 in hippocampus were decreased in APP/PS1 double transgenic mice, but increased in concentration-dependent manner in curcumin treatment group,especially in CA1 and CA3 region. MAP2 may play an important role in the generation or development of AD. MAP2 were injured in APP/PS1 double transgenic mice while curcumin could resume the expression of MAP2. Further research is needed to provide mechanism for this result.

Abstract: The influence of ultrasound-microwave during extraction of Ricinus communis(L）allergen from castor bean meal (CBM) was investigated. The method was successfully applied for the ultrasound- microwave assisted extraction (UME) of allergen from CBM. The corresponding extraction parameters including the extraction temperature, ultrasonic-microwave power and extraction time were investigated. The CBM was crushed, sifted, ground and suspended for 4 h in 25% ethanol water (1:4) at room temperature followed by ultrasound-microwave. The yield, polysaccharose and protein contents were determined after each processing step. The purity of extracted allergen was tested through HPLC and the Sephadex G-100 and Tricine-SDS-PAGE detection with unique electrophoresis strip documented that allergen was purifed.. The paper describes the conditions of the temperature,time,and microwave-power for the method reliability with UME and suitablilty for the effective extraction of allergen from CBM.

Abstract: In this study, we aimed to investigate the effects and mechanism of Genistein on proliferation inhibition and onset of apoptosis in ovarian carcinoma cell line 3AO. Antiproliferative effect of Genistein against 3AO was tested by MTT method. Apoptotic phenotype of 3AO was observed by electron microscope. Cell apoptosis percentage and cell cycle phase distribution of 3AO were measured by flow cytometric assay. The expression of proliferation and apoptosis associated protein was determined by using immunocytochemical method. A time-dependent and dose-dependent proliferation inhibition was demonstrated in 3AO, and Genistein induced a G₂/M cell cycle arrest . The characteristic morphological changes of apoptosis in 3AO cells were observed after treated by Genistein. 20μmol/L Genistein could down-regulate the expression of PCNA, CyclinB₁ and Bcl-2; and up-regulate the expression of P21^WAF1/CIP1 and Bax. The time-dependent and dose-dependent proliferation inhibitory effect of genistein on ovarian cancer appears to be due to the up-regulation of P21^WAF1/CIP1 and PCNA expression, down-regulation of CyclinB₁ expression, and the apoptosis is related to the up-regulation apoptotic proteins Bax and down-regulation of anti-apoptotic proteins Bcl-2. The results provide evidence for the potential usefulness of Genistein in the prevention and treatment of human ovarian carcinoma.

Abstract: Extracting flavone from bamboo leaves by ultrasonic, the flavonoid extraction conditions was optimized by single factor experiment and orthogonal experiment design, and the traditional heat reflux method was compared. The results showed that: the optimum process conditions of ultrasonic wave extraction of flavonoids from bamboo leaves were ethanol concentration of 70%, solid to liquid ratio of 1:20, extracting 20 min in 60 °C. Under this condition, extraction content of total flavonoids was up to 4.538 mg.g^-1. And ultrasonic assisted extraction was superior to hot reflux extraction.