Animals ; Brain-Derived Neurotrophic Factor ; genetics ; Cells, Cultured ; Female ; Gene Expression ; drug effects ; Neurons ; drug effects ; Phenylalanine ; toxicity ; Phenylketonurias ; complications ; Rats ; Rats, Sprague-Dawley ; rho GTP-Binding Proteins ; genetics

The human canstatin cDNA was amplified by RT-PCR and then directionally cloned into pU? expression vector. The recombinant pU?-Can vector was connected with the screening marker (bar box), to construct a eukaryotic expression vector called pU?-Can-Bar. This expression vector was introduced into the D.salina by glass beads method. The screening culture of transformants of D.salina was performed in solid media containing 5 ?g/ml PPT, and the analyses of transformants were carried out through PCR and Southern blot. PCR results revealed that specific 700 bp products were detected in the different transformants of D.salina but not in negative control. Southern blot analysis further demonstrated that human canstatin gene was integrated into the D.salina genome. Moreover, the results of genetic stability analyses of transformants demonstrated that canstatin gene was stably inherited in the D.salina transformants. The successful preparation of the D.salina transformants will provide the experimentation evidence for producing canstatin protein cosmically by using the D.salina bioreactor and give a better prophase work basis for clinic application of canstatin protein early.

Pancreatic ductal adenocarcinoma (PDAC) is one of the five most malignant cancer. ZX-1201 is one of the active constituents in Alismatis Rhizoma,a well-known traditional Chinese medi-cine with a wide variety of pharmacological properties including diuretic,anti-hyperlipidemic,anti-atheroscle-rotic,anti-cancer,anti-inflammatory and anti-oxidative activities.We investigated the inhibitory effect of ZX-1201 on pancreatic cancer and the relevant molecular mechanism in vitro and in vivo. ZX-1201 inhibited the growth and metastasis of PANC-1 cells in BALB/c nude mice significantly.ZX-1201 inhibited the function of AQP1 via directly interaction and involved in the reversion process of ZX-1201 on TGF-β1. CTGF was an important protein in the reversion process of ZX-1201 on TGF-β1.ZX-1201 inhibited the migration of PANC-1 and CPFAC-1 cells induced by TGF-β1in vitro.ZX-1201 reversed the down-regu-lated of epithelial markers and up-regulated of mesenchymal markers, as well as the up-regulated of Snail and p-Smad2/3 induced by TGF-β1.And ZX-1201 reversed Epithelial-Mesenchymal Transition by down-regulating AQP1 and inhibiting translocation of β-catenin, the promotor of CTGF. According to these,ZX-1201 inhibited the migration of pancreatic cancer cells.We concluded that ZX-1201 inhibited the growth and metastasis of PANC-1 cells in vivo significantly.And AQP1,β-catenin and CTGF were the pivotal proteins in the process of ZX-1201 inhibiting PANC-1 cells migration induced by TGF-β1.

OBJECTIVETo evaluate the efficacy and safety of a phenylalanine-free amino acid-based enteral formula (AA-PKU2) in the treatment of children with phenylketonuria (PKU) aged 1-8 years.

METHODSA prospective, open, self-controlled, multi-center trial was performed, enrolling 121 PKU children (1-8 years in age) consecutively between July, 2009 and May, 2011. Enteral nutrition therapy was administered for 32 weeks. The data on blood phenylalanine (PHE) levels, metal development, weight, height, head circumference, serum nutritional biomarkers (total protein, pre-albumin, albumin, total cholesterol, total triglyceride, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol), and measurements from routine blood and urine examinations and from renal and hepatic function tests were collected before the therapy and at 8 weeks and 32 weeks after the therapy and were comparatively analyzed.

RESULTSThe mean blood PHE level at 8 and 32 weeks of AA-PKU2 treatment was 353±253 and 361±280 µmol/L respectively, significantly lower than that before the treatment (487±327 µmol/L; P<0.01). The difference in intelligence quotient scores before and after AA-PKU2 treatment was not significant (P>0.05) when assessed by the Gesell tests in children aged 1-4 years but significant (P<0.01) when assessed by WPPSI or WISR-R tests in children over 4 years. The average height, weight and head circumference at 8 and 32 weeks after treatment were significantly increased as compared to these measurements before treatment (P<0.01) with absolute levels similar to those in the control children. In contrast, the mean values of total protein, pre-albumin, albumin, total cholesterol, total triglyceride, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol at both time points were not different either from those prior to the treatment or from those in the control children. Mild diarrhea was the adverse events associated with AA-PKU2 treatment, which occurred in 3 (2.5%) cases. All these 3 patients fully recovered without treatment.

CONCLUSIONSThe phenylalanine-free amino acid-based formula, AA-PKU2, is effective and safe in controlling blood PHE levels and improving mental development with adequate nutritional support in PKU.

Child ; Child, Preschool ; Enteral Nutrition ; Female ; Humans ; Infant ; Intelligence ; Male ; Phenylalanine ; blood ; Phenylketonurias ; diet therapy ; psychology ; Prospective Studies

Gardeniae Fructus contains volatile ingredients, however, the species and proportions in different processed products of Gardeniae Fructus are different. In this experiment, volatile ingredients were separated by steam distillation with content of 1.2, 1.0, 0.9, 0.7 µL · g(-1) in Gardeniae Fructus, fried Gardeniae Fructus, stir-baked Gardeniae Fructus, Gardeniae Fructus fried into carbon respectively. One hundred and twenty-four kinds of volatile components were identified by GC-MS. Fifty-three kinds of volatile ingredients consisted in Gardeniae Fructus accounting for 93.85%, 54 kinds in fried Cardeniae Fructus accounting for 92.01%, 32 kinds in stir-baked Cardeniae Fructus accounting for 91.59% and 43 kinds in Gardeniae Fructus fried into carbon accounting for 90.81%. In this paper, analysis of Gardeniae Fructus by GC-MS provides a scientific basis for elucidating the mechanism of different processed products.
Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; Gardenia ; chemistry ; Gas Chromatography-Mass Spectrometry ; Molecular Structure ; Volatile Organic Compounds ; chemistry

Processed Chinese medicine is the core of traditional Chinese medicine (TCM) industry chain,which directly affects the clinical efficacy and. safety of Chinese patent medicine and clinical formula Decoction pieces. Studied the variation of effective substance in vivo Chinese medicine processing before and after processed, clarifying the effective substance and processing principle is a top priority of the development of Chinese medicine processing. The traditional research method chiefly focus on the variation about chemicals in vitro of processed Chinese medicine, it cannot reveal that the integrity and complexity of processed Chinese medicine efficacy changes, so the change process is the focus of future research in vivo on the base of effective substance of TCM This paper described the research on the base of effective substance of TCM and Processed Chinese medicine research status in vitro, discussed the analytical methods (plasma chemistry, pharmacokinetics, metabonomics) of the dynamic process in vivo about processed Chinese medicine, and pointed out development and related problems in process in vivo on the base of effective substance of TCM, which could provided research ideas and methods for in-depth interpretation of Chinese medicine processing mechanism.
Animals ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Humans ; Medicine, Chinese Traditional

Objective To study CD35on erythrocytes and erythrocyte chemokine recepter(ECKR)in patients with psoriasis in order to explore red blood cell(RBC)innate immune function and its possible role in the pathogenesis of psoriasis.Methods The rapid natural immune reaction on cancer cells was measured with RBCs isolated from fresh blood.Expression of CD35on erythrocytes was detected by flow cy-tometry.The level of IL-8,which was bound to isolated erythrocytes,was measured by enzyme linked im-munosorbent assay(ELISA)in the supernatant after centrifugation,which represented ECKR binding activity.Results Rosette formation rates of RBC adhering to cancer cells and expression of CD35on RBCs were significantly higher in psoriatic patients than those in control group(P

Objective To investigate the levels of serum vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in patients with non-small cell lung cancer(NSCLC) and relationships with c1inicopatho1ogica1 characteristics and their clinical significance. Methods The concentrations of serum VEGF and bFGF were detected by enzyme-linked immunosorbent assay(ELISA) in 40 patients with NSCLC before and after chemotherapy. Results The level of serum VEGF in patients with Ⅳ stage NSCLC was significantly higher than that of Ⅲ stage(P

Recent studies indicate that beta-amyloid (Abeta) is the key factor to cause neuronal degeneration in Alzheimer's disease (AD). In the present study, we set up an Abeta induced PC12 cell damage modle and studied the protective effect and related mechanisms of T(10), monomer extracted from Chinese herb Tripterygium wilfordii Hook F. PC12 cells were treated with different concentrations of Abeta (5x10(-4), 5x10(-3), 5x10(-2), 5x10(-1), 5, 50 micromol/L) for 48 h, cell viability was detected by MTT conversion. The apoptotic rate of PC12 cells was quantitatively determined using FACS assay. After PC12 cells were treated with 1x10(-11) mol/L T(10) for 48 h and then co-treated with 50 micromol/LAbetafor 48 h, the apoptotic rate and the change in intracellular Ca(2+) concentration of PC12 cells were analyzed by FACS assay and confocal, respectively. It was found that 5 micromol/L Abeta decreased the cell viability to 66.3% and 50 micromol/L Abeta decreased it to 55.1%, significantly different from that of the control group. After treatment with 50 micromol/L Abeta for 48 h, the apoptotic rate of PC12 cells increased obviously. The apoptotic rate was 5.37% in the control group, while after treatment with 0.5, 5 and 50 micromol/L Abeta for 48 h, the apoptotic rate of PC12 cells went up to 10.19%, 8.02% and 16.63%, respectively. At the same time, the concentration of intracellular Ca(2+) increased greatly after treatment with 50 micromol/L Abeta for 48 h. At the concentration of 1x10(-11) mol/L T(10) remarkably inhibited the apoptosis induced by 50 micromol/L Abeta. In the naive group, the apoptotic rate was 4.83%. The apoptotic rate went up to 17.24% after treatment with 50 micromol/L Abeta for 48 h. After co-treatment with 1x10(-11) mol/L T(10) and 50 micromol/L Abeta, the apoptotic rate decreased to 8.91%, significantly different from that of the control group. At the same time, at the concentration of 1x10(-11 )mol/L T(10) remarkably inhibited the increase of intracellular Ca(2+) concentration induced by Abeta. The results indicate that T(10) has obvious protective effect on PC12 cells, which may be related to the inhibition of the cell apoptosis and increment of intracellular Ca(2+) concentration induced by Abeta.
Alzheimer Disease ; pathology ; Amyloid beta-Peptides ; toxicity ; Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Diterpenes ; pharmacology ; Epoxy Compounds ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; Peptide Fragments ; toxicity ; Phenanthrenes ; pharmacology ; Rats ; Tripterygium ; chemistry

Adult ; Antarctic Regions ; Emergency Medical Services ; statistics & numerical data ; Expeditions ; Female ; Humans ; Male ; Middle Aged ; Patient Care Team ; statistics & numerical data ; Patient-Centered Care ; statistics & numerical data ; Retrospective Studies