OBJECTIVEOptimize the technic condition of Duzhong Jiangya prescription with Semi-bionic Extraction.

METHODUsing homogeneous design, under the same materials granularity, decoction temperature, consumption of water, filtration, concentration, and taking aucubin, hydrochloric acid stachydrine, maloicacid, baicalin, ethanol extract, dry extract as the indexes, the results were comprehensive considered to optimize the semi-bionic extraction condtions.

RESULTThe optimized SBE extraction conditions are the following: pH values of the water for the thrice extraction is 5.845 3, 7.496 1, 8.011 7, and the total extraction time is 3.418 0 h.

CONCLUSIONConsidering the fact of manufacture, the SBE extraction conditions are the following: pH values of the water for the thrice extraction is 6.0, 7.5, 8.0, and the thrice extraction time is 1.5, 1.0, 1.0 h.

Antihypertensive Agents ; chemistry ; isolation & purification ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eucommiaceae ; chemistry ; Flavonoids ; analysis ; Glucosides ; analysis ; Hydrogen-Ion Concentration ; Iridoid Glucosides ; Iridoids ; analysis ; Leonurus ; chemistry ; Plants, Medicinal ; chemistry ; Proline ; analogs & derivatives ; analysis ; Prunella ; chemistry ; Scutellaria baicalensis ; chemistry ; Technology, Pharmaceutical ; methods ; Time Factors ; Triterpenes ; analysis ; Uncaria ; chemistry

OBJECTIVETo explore the effect of Bushen Tiaojing Recipe (BTR) on the quality of oocytes, reproductive hormones, and the expression of bone morphogenetic protein-15 (BMP15) of in vitro fertilization-embryo transfer (IVF-ET) patients.

METHODSSixty infertility patients who prepared for IVF-ET were assigned to two groups according to the treatment order, the treatment group [20 cases, treated with BTR + controlled ovarian hyperstimulation (COH)] and the control group (treated with COH alone, 40 cases). Age, the time limit for infertility, basal follicle-stimulating hormone (bFSH) concentration, usage days and the dosage of gonadotropins (Gn), serum levels of estradiol (E2), luteotropic hormone (LH), and progesterone (P) on the HCG injection day, the number of retrieved occytes, the fertilization rate, the number of embryos, the high quality embryo rate, and the clinical pregnancy rate were compared. Concentrations of follicle-stimulating hormone (FSH), LH, E2, testosterone (T), and P in the follicular fluid were detected via chemiluminescence microparticle immunoassay. The mRNA and protein expression of BMP-15 in mature granulosa cells was detected by real-time fluorescent PCR and Western blot.

RESULTSThirty-two patients were pregnant and the total pregnancy rate was 53.3%. Of them, 19 were pregnant and the total pregnancy rate was 47.5% in the control group, while 20 were pregnant and the total pregnancy rate was 65.0% in the treatment group. But there was no statistical difference between the two groups (P > 0.05). Compared with the control group, the Gn dosage was lower and the high quality embryo rate was higher in the treatment group, showing statistical difference (P < 0.05). There was no statistical difference in serum concentrations of E2, LH, or P on the HCG injection day, the number of retrieved oocytes, or the fertilization rate (P > 0.05). Compared with the control group, FSH concentrations in the follicular fluid were significantly lower and LH concentrations were significantly higher in the treatment group (P < 0.05). The LH concentrations in the follicular fluid were significantly higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).There was no statistical difference in E2, T, or P concentrations (P > 0.05). The mRNA and protein expression of BMP-15 in granulosa cells was higher in the treatment group than in the control group (P < 0.05). It was also higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).

CONCLUSIONDuring the IVF-ET process, BTR could elevate the quality of oocytes, and increase the sensitivity of ovarian follicles to exogenous Gn, which was correlated with the mRNA and protein expression of BMP-15 in granulosa cells, and changing concentrations of FSH and LH.

Adult ; Bone Morphogenetic Protein 15 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Embryo Transfer ; Estradiol ; blood ; metabolism ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; metabolism ; Follicular Fluid ; metabolism ; Humans ; Luteinizing Hormone ; blood ; metabolism ; Oocytes ; drug effects ; Pregnancy ; Progesterone ; blood ; metabolism ; Testosterone ; metabolism ; Young Adult

Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.

Objective To discuss effect on prevention of postpartum hemorrhage of caesarean section by us- ing calcium gluconate combined with oxytocin and misoprostol.Methods 385 cases of caesarean section were select- ed and randomized into O(Oxytocin) group and OM(Oxytocin+ Misoprostol) group and COM (Calcium gluconate+ Oxytocin+Misoprostol)group.Results The mean operative blood loss in O group and OM group and COM group were (300?50.24)ml,(220?30.83) ml,(150?45.52) ml.The amount of the mean operative blood loss of COM group was significantly lower than those of O group and OM group(P＜0.05).The amount of bleeding of 2 hours after delivery in O group and OM group and COM group were (400?45.52)ml,(260?60.43)mi and(210?50.54) ml.The amount of bleeding of COM group was significantly lower than those of O group and OM group (P＜0.05).Conclusion The prevention by used calcium gluconate combined with oxytocin and misoprostol is efficient in reducing the amount of postpartum hemorrhage of caesarean section.The operation of medicine is easy and safe and economic.

Background The underlying mechanism of choroidal neovascularization(CNV) is multifactorial and complex.Focal adhesion kinase(FAK) plays a crucial role in controlling essential cellular processes and influencing distinct steps of the angiogenic response.But to our knowledge,seldom study on the effect of FAK on CNV formation has been reported previously.Objective In this study,the effect of several CNV risk factors on the expression of FAK in cultured retinal pigment epithelium(RPE) cells was investigated to illuminate effect of FAK on CNV.Methods Human RPE cells were isolated from donor eyes and exposed to H2O2,swallow of outer segment of photoreceptors(POS) and extracellular matrix(ECM) separately with the treating as follows:RPE cells were co-cultured with 10,20,50 and 100μmol/L H2O2 for 20 days;POS(1×106/ml) were co-cultivated with RPE cells for 20 days(setting control group,POS group,hypoxia group with 200μmol/L CoCl2,and POS+hyoxia group);RPE cells were cultured on the plates coated with 100mg/L fibronectin(FN),laminin(LN) or collagen typeⅠfor 30minutes or 1 hour.The expression of FAK and pFAK in RPE cells were examined by Western blot analysis.Results FAK was highly expressed in the 20μmol/L and 50μmol/L H2O2 groups compared with control group(P＜0.01);while he expression level of pFAK was reduced after treated with H2O2 in comparison with the control group(P＜0.01).After cultured with POS for 20 days,the undigested lysosome could be observed in RPE cells.The expressions of FAK and pFAK in RPE cells were not significantly changed between control group and POS groups(P＞0.05),but those in hypoxia group were significantly up-regulated in comparison with control group(P＜0.01).Compared with the hypoxia group,the expression amount of pFAK was elevated in POS+hyoxia group(P＜0.01).In comparison with control group,the increased pFAK expression was seen in FN,LN and collagen typeⅠtreating for 1-hour groups(P＜0.05,P＜0.01).Conclusion FAK pathway participates in several CNV-initiated signaling,such as H2O2,POS and ECM,in cultured RPE cells.It is reasonable to believe that FAK potentially plays an important role in CNV-dependent disorder.

Objective To investigate the relationship between serum adiponectin and metabolic syndrome(MS) in children and adolescents with obesity and nonalcoholic fatty liver disease(NAFLD).Methods Four elementary schools and 4 middle schools were selected from Haidian district in Beijing with representative cluster sampling.Two hundred and eighty obese children(obese group),65 obese children with NAFLD(NAFLD group) and 264 normal weight children(healthy control group) aged 7 to 18 years were recruited from the 8 schools with uncompletely randomized sampling.Data including questionnaire,anthropometric measurements,B type ultrasonographic examination for liver were collected and fasting blood laboratory assay were determined.Variables including triglyceride(TG),adiponectin,alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were skewed distribution and natural logarithmical transformations were performed.Chi-square test for category and multiple binary Logistic regression analysis were used to statistical analysis.Results Body mass index(BMI) and waist circumference(WC) in obese group and NAFLD group were higher than those in healthy control group.All the chi-square tests for trend among the 3 groups were statistically significant(P

Polysaccharide sulfate (PSS) is a new type of antiatherosclerotic medicine for its effects of anticoagulation, anti-thrombosis and modulation of dyslipidemia. However, it is still uncertain whether PSS could modulate the diabetic dyslipidemia or not. Here, the rat model of diabetic dyslipidemia was developed and the effects of PSS on glucose and lipid levels were investigated in this animal model. Wistar rats were iv injected with streptozotocin 20 mg x kg(-1) after feeding with high fat diet for one and a half month. Then, rats received orally PSS (30, 90, and 180 mg x kg(-1)) for 1 month. After oral treatment with PSS (90 and 180 mg x kg(-1)) for 1 month, the levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) were significantly reduced and the level of high density lipoprotein-cholesterol (HDL-C) increased, compared with diabetic control rats. Moreover, PSS (30, 90, and 180 mg x kg(-1)) had a tendency to reduce glucose and insulin levels, and significantly increased insulin sensitivity index. Our results suggest that PSS could improve insulin sensitivity and relieve dyslipidemia in diabetic dyslipidemic rats.
Administration, Oral ; Animals ; Blood Glucose ; metabolism ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Diabetes Mellitus, Experimental ; blood ; chemically induced ; complications ; Dyslipidemias ; blood ; etiology ; Hypolipidemic Agents ; administration & dosage ; pharmacology ; Insulin ; blood ; Insulin Resistance ; Male ; Polysaccharides ; administration & dosage ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Streptozocin ; Sulfates ; administration & dosage ; pharmacology ; Triglycerides ; blood

This paper is aimed to study the effects of nitrogen form on the growth and quality of Chrysanthemums morifolium at the same nitrogen level. In order to provide references for nutrition regulation of Ch. morifolium in field production, pot experiments were carried out in the greenhouse at experimental station of Nanjing Agricultural University. Five proportions of ammonium and nitrate nitrogen were set up and a randomized block design was applied four times repeatedly. The results showed that the growth and quality of Ch. morifolium were significantly influenced by the nitrogen form. The content of chlorophyll and photosynthesis rate were the highest at the NH4(+) -N /NO3(-) -N ratio of 25:75; The activities of NR in different parts of Ch. -morifolium reached the highest at the NH4(+) - N/NO3(-) -N ratio of 0: 100. The contents of nitrate nitrogen in the root and leaves reached the highest at the NH4(+) -N/NO3(-) -N ratio of 50:50. The activities of GS, GOGAT and the content of amylum increased with the ratio of NO3(-) -N decreasing and reached it's maximum at the NH4 + -N/NO3 - -N ratio of 100: 0. The content of ammonium nitrogen were the highest at the NH4 + -N /NO3 --N ratio of 75: 25, while the content of soluble sugar reached the highest at the NH4(+)-N/NO3(-) -N ratio of 25: 75. The content of flavones, chlorogenic acid and 3,5-O-dicoffeoylqunic acid were 57.2 mg x g(-1), 0.673% and 1.838% respectively, reaching the maximum at the NH4(+) -N /NO3(-) -N ratio of 25:75; The content of luteoloside increased with the ratio of NO3(-) -N increasing and reached it's maximum at the NH4(+) -N/NO3(-) -N ratio of 0: 100. The yield of Ch. morifolium reached it's maximum at the NH4(+) -N /NO3(-) -N ratio of 25:75. Nitrogen form has some remarkable influence on the nitrogen metabolism, photosynthesis and growth, Nitrogen form conducive to the growth and quality of Ch. morifolium at the NH4(+) -N /NO3(-) -N ratio of 25: 75.
Ammonium Compounds ; metabolism ; pharmacology ; Chlorophyll ; metabolism ; Chrysanthemum ; drug effects ; growth & development ; metabolism ; Flowers ; drug effects ; growth & development ; metabolism ; Glutamate Synthase ; metabolism ; Glutamate Synthase (NADH) ; metabolism ; Glutamate-Ammonia Ligase ; Nitrates ; metabolism ; pharmacology ; Nitrogen ; metabolism ; pharmacology ; Photosynthesis ; drug effects ; Plant Leaves ; drug effects ; growth & development ; metabolism ; Plant Proteins ; metabolism ; Plant Roots ; drug effects ; growth & development ; metabolism ; Plant Stems ; drug effects ; growth & development ; metabolism

OBJECTIVETo introduce green fluorescence protein (GFP) into E. coli O157:H7 and improve the detection methods for this bacteria, and to study the kinetics of E. coli O157:H7.

METHODSThe plasmid pGFP was transferred into E. coli O157:H7. The characteristic of the new built O157:H7-pGFP strain was evaluated. Some food samples were inoculated with the recombinant strain under certain temperature to imitate different storage circumstances. The contaminated E. coli O157:H7 were counted after certain time.

RESULTSThe pGFP was stable in E. coli O157:H7. The E. coli O157:H7-pGFP inoculated in ground poultry meat and pasteurized milk were enriched to 35 000 approximately 200 000 times in 12 h under higher storage temperature (37 degrees C), whereas the quantity decreased slowly under lower temperature (4 degrees C).

CONCLUSIONThe recombinant strain with the characters of ampicillin resistance and green fluorescence under UV 365 nm was a useful tool in detection methods improvement and bacteria survival studies.

Colony Count, Microbial ; Escherichia coli O157 ; isolation & purification ; Food Contamination ; Food Handling ; methods ; Food Microbiology ; Green Fluorescent Proteins ; Meat Products ; microbiology

OBJECTIVETo prepare monoclonal antibodies (MAb) and antisera specific for Escherichia coli (E. coli) O157, and to develop a sandwich enzyme-linked immunosorbent assay (ELISA) to detect E. coli O157 in foods.

METHODSSpleen cells from BALB/c mice immunized with the somatic antigen of E. coli O157:H7 were fused with murine Sp2/0 myeloma cells. The hybridoma cell line specific for E. coli O157 was established after having been subcloned. Antisera specific for E. coli O157 was prepared by intravenous injection into New Zealand rabbits with a stain of E. coli O157:H7. The sandwich ELISA was developed with the polyclonal antibody as the capture antibody and the MAb 3A5 as the detection antibody. The inoculated ground poultry meat and pasteurized milk were tested to confirm efficiency of the method.

RESULTSMAb 3A5 specific for E. coli O157 and O113:H21 belonged to subtype IgM. The ascetic titers of the antibody was 1:1x10(6). No cross-reactivity of the MAb was observed with strains of Salmonella spp, Yersinia enterocolitica, Shigella dysenteriae, etc. The purified polyclonal antibody had a titer of 1:1x10(5) with E. coli O157. The detection limit of this sandwich ELISA was 10(3)-10(4) cfu E. coli O157/mL in pure culture with a high specificity, which was characterized by every non-O157 strain with negative response. With 10h enrichment procedure, E. coli O157:H7 recovered well from inoculated ground poultry meat and pasteurized milk at levels of 0.1 cfu/g and 0.1 cfu/mL.

CONCLUSIONMAb 3A5 specific for E. coli O157 and O113:H21 can be produced by immunizing BALB/c mice with a strain of E. coli O157:H7. Then a sandwich ELISA can be developed with the polyclonal antibody as the capture antibody and the MAb 3A5 as the detection antibody. The method is proved to be a sensitive and specific technique to detect low number of E. coli O157 in food.

Animals ; Antibodies, Monoclonal ; immunology ; Antigens, Bacterial ; immunology ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli O157 ; immunology ; isolation & purification ; Female ; Food Microbiology ; Mice ; Mice, Inbred BALB C ; Milk ; microbiology ; Poultry ; microbiology ; Rabbits