SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.

In the current study, nine flavonoids were isolated and purified from 75% ethanol extract of Selaginella uncinata (Desv.) Spring by column chromatographic techniques over macroporous resin, polyamide, silica gel, Sephadex LH-20 and pre-HPLC. On the basis of their physico-chemical properties and spectroscopic data analyses, these compounds were elucidated as cirsimarin (1), nepitrin (2), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), apigenin-6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (4), apigenin-7-O-β-D-glucopyranoside (5), 2,3-dihydroamentoflavone (6), 4'-O-methylamentoflavone (7), 2,3-dihydro-4'-O-methyl-amentoflavone (8), and 2,3,2",3"-tetrahydron-4'-O-methyl-robustaflavone (9). Compounds 1-5 belong to flavonoid glycosides and were isolated from the genus Selaginella for the first time.
Flavonoids ; analysis ; Selaginellaceae ; chemistry

OBJECTIVETo establish a HPLC-based method for simultaneous determination of 2 classes of compounds (flavonoids and chromones) and 6 their effective components,(including prin-O-glucosylcimifugin, cimifugin, 4'-O-beta-D-glucosyl- 5-O-methylvisamminol, quercetin, sec-o-glucosylhamaudol and formononetin), in Yupingfeng Decoction.

METHODSHPLC-based separation of the agents was performed on Agilent Extend-C(18) column (4.6 mm x 250 mm, 5 microm) at 25 degrees with the mobile phase of MeOH-1% acetic acid water solution (gradient elution), flow rate of 0.8 ml/min and detection wavelength of 254 nm.

RESULTS AND CONCLUSIONHPLC allowed simultaneous quantitative determination of the 6 components in Yupingfeng Decoction, and they showed good linear relationships when their sample amount ranged 90-1810 ng, 97-1940 ng, 190-1906 ng, 105-3144 ng, 88-2625 ng and 109-3279 ng, respectively, with correlation coefficients all beyond 0.9999 and average recovery rates of 98.2%, 99.1%, 97.3%, 97.8%, 98.8% and 99.2%, respectively. This simple and convenient method accommodated a broad linear range with high sensitivity and precise and reproducible results.

Chromatography, High Pressure Liquid ; methods ; Chromones ; analysis ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; Isoflavones ; analysis ; Quercetin ; analysis ; Reproducibility of Results

HIV-1 integrase (IN) is an essential enzyme for retroviral replication. There is no analogue for this enzyme in human cells so that inhibition of IN will not bring strong effect on human body. Thus, HIV-1 IN has become a rational target for therapy of AIDS. This review provides a comprehensive report of alpha, gamma-diketo IN inhibitors discovered in recent years. Compilation of such data will prove to be beneficial in developing QSAR, pharmacophore hypothesis generation and validation, virtual screening and synthesis of compounds with higher activity.
Anti-HIV Agents ; chemical synthesis ; chemistry ; pharmacology ; HIV Integrase ; chemistry ; physiology ; HIV Integrase Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; HIV-1 ; drug effects ; Humans ; Keto Acids ; chemical synthesis ; chemistry ; pharmacology ; Molecular Structure ; Quantitative Structure-Activity Relationship

Objective To investigate the clinical significance of serum cytokines concentrations and A- PACHE scores in evaluating the illness state for critical trauma patients.Methods A clinical prospective self-control trial was performed,in which 36 patients admitted to ICU by SIRS were enrolled.Objects were divided into mild and severe trauma group according to APACHE score.The TNF-?and IL-6 concentrations were determined on the 1st, 3rd and 5th day of admission,the APACHE score were assessed at the same time.Statistic analysis was performed according to this group.Results The TNF-?concentrations decreased continuously in the following days while IL-6 decreased from the 7th day in the mild trauma group.In the severe trauma group the TNF-?and IL-6,APACHE score concentrations kept increasing.There was a significant difference of TNF-?and IL-6 concentrations between severe trauma and mild trauma group.Conclusion Dynamic measurement of TNF-?and IL-6 concentrations with APACHE score provide great help to evaluate the illness state and predict the prognosis.

Intestinal tract, which produces more than fifty kinds of gut peptides, is regarded as the largest endocrine organ. With regard to the gut peptides, a number of studies were focused on their structure, function and the roles in some diseases. The changes in output or distribution of gut peptides in the intestinal tract during development have been largely unknown. This study was aimed to investigate the changes of somatostatin (SST) and somatostatin receptor 2 (SSTR2) in small intestinal and hepatic tissues during the development of macaque. The tissue samples of small intestine, liver or blood samples from peripheral and portal vein of 4 macaques in 6-month fetus, 2-day neonate, 45-day neonate and adult were obtained after anesthetization. The concentrations of SST in blood or tissues of macaques were measured by radioimmunoassay. The distributions of SST in small intestinal or hepatic tissues were visualized by immunohistochemical staining. The expression of SSTR2 was detected by in situ hybridization. SST concentration of intestinal tissue in 6-month-old macaque was (27.3+/-16.6) ng /mg protein and light positive staining of SST was localized in mucosal crypts but negative in muscle layer. The intestinal concentration of SST increased gradually with macaque development and reached to the peak [(120.1+/-35.3) ng /mg protein] in adult. It was significantly higher than that in fetus (P<0.01). Strong positive staining of SST was found in both mucosal crypts and myenteric nerve plexus of adult animal. SSTR2 was obviously expressed in intestinal epithelium of fetus but its expression was greatly reduced in epithelium and was shifted to mucosal crypts when grown to adult. Negative staining of SSTR2 in muscle layer of fetal or neonatal macaque turned to be positive in myenteric nerve plexus of adult. The levels of SST or SSTR2 in liver decreased gradually during development. SST concentrations of small intestinal tissue kept significantly higher than those of hepatic tissues in the macaque developing stages. SST levels of portal vein were also maintained significantly higher than those of peripheral blood in the macaque developing stages. In conclusion, the level of SST and expression of SSTR2 in mucosal crypt increased gradually with macaque development. SST from intestinal tract was quickly degraded in portal vein before entering into liver. SST positive myenteric nerve plexus was visualized only in mature macaque.
Animals ; Animals, Newborn ; Fetus ; Intestine, Small ; metabolism ; Liver ; metabolism ; Macaca mulatta ; growth & development ; metabolism ; Male ; Receptors, Somatostatin ; metabolism ; Somatostatin ; metabolism

OBJECTIVETo identify the localization of hair follicles stem cell (HFSC) in different stages of hair and explore the differentiating capacity of HFSC into epidermis in vitro.

METHODSHFSC were detected by K19 immunostaining in normal human skin. Then, the isolated HFSC through enzyme digestion were seeded on dermal equivalent (DE) and cultured between the air-liquid interfaces for 14 days. Skin-equivalents was harvested and used for evaluation.

RESULTSHFSC mainly located in outer root sheet in hair follicle and human anagen hair follicles containing two distinct reservoirs for K19-positive cells located in the bulge and bulb of the follicle. These two reservoirs fused in line of outer root sheets during the catagen-telogen transition phase and individualized again in the newly forming anagen hair follicle. Based on DE, growing HFSC built a multilayered and confined epidermis.

CONCLUSIONHFSC located in outer root sheets can promote hair cycle and differentiate into epidermis in vitro.

Cell Differentiation ; physiology ; Cells, Cultured ; Epidermis ; cytology ; Hair Follicle ; cytology ; Humans ; Stem Cells ; cytology

OBJECTIVETo set up the HPLC fingerprint of Glycyrrhiza uralensis in decoctions prepared from various combinations of the recipe named Mahuang decoctions and study the influence of compatibility on fingerprint of G. wralensis in decoctions.

METHODThe RP-HPLC method was used, chromatography conditions were Zorbax SB-C18 column (4.6 mm x 250 mm, 5 microm), the acetonitrile-1.2% acetic acid (gradient elution) as mobile phase and detective wavelength at 254 nm.

RESULTThe method is stable and reliable with a good reproducibility and provides a reference standard for the quality control of G. uralensis, it can be used to determine the fingerprint of G. aralensis in Mahuang decoctions. The result showed that 8 peaks were common in the fingerprint of G. uralensis in Mahauang decoctions. The area changes of 8 common peaks in decoctions prepared from various combinations of Mahuang decoctions were studied.

CONCLUSIONThe fingerprint of G. uralensis in Mahuang decoctions is markedly influenced by herba ephedrae and ramulus cinnamomi and semen armeniacae amarum. The areas of 7 common peaks were reduced obviously, while the relative areas of 6 common peaks were increased remarkably.

Chromatography, High Pressure Liquid ; methods ; Cinnamomum ; chemistry ; Drug Combinations ; Drug Interactions ; Drugs, Chinese Herbal ; chemistry ; Ephedra sinica ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Plants, Medicinal ; chemistry ; Prunus ; chemistry ; Quality Control ; Reproducibility of Results

Gegen Qinlian Decoction is a classical prescription with the function of relieving exterior and interior syndromes.The formula contains complex chemical components and is prepared into several dosage forms regulating the function of the gut.The review discusses the chemical components,common dosage forms,clinical application and effect on the intestinal barrier of Gegen Qinlian Decoction,so as to provide a basis for its further development and utilization.

Adult ; CD5 Antigens ; metabolism ; Carcinoma, Papillary ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Choristoma ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Hamartoma ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-kit ; metabolism ; Thymoma ; metabolism ; pathology ; Thymus Gland ; pathology ; Thymus Neoplasms ; metabolism ; pathology ; Thyroid Neoplasms ; metabolism ; pathology ; surgery ; Thyroidectomy ; methods