Aim To investigate whether the pain modi-fication by group I metabotropic glutamate receptors (mGluRs)required the involvement of Src homology-2 domain-containing phosphatase-2 (SHP-2 ).Methods Co-immunoprecipitation was performed to examine the possible interaction between SHP-2 and group I mGluRs in spinal cord dorsal horn of mice.By measur-ing the paw withdrawal thresholds,the effects of SHP-2 inhibitor NSC-87877 or its catalytically inactive SHP-2 (C459S ) mutant on allodynia induced by group I mGluRs agonist DHPG (50 nmol)were observed.Re-sults Anti-mGluR5 antibody was able to co-immuno-precipitate SHP-2 from spinal dorsal horn of mice, while no SHP-2 was precipitated by anti-mGluR1 anti-body.Inactivation of SHP-2 by NSC-87877 (6 nmol) or SHP-2 (C459S ) effectively attenuated allodynia caused by DHPG.Conclusion SHP-2 can physically interact with mGluR5.The activation of SHP-2 may be necessary for group I mGluRs to process the nocicep-tive information.

Objective To optimize and establish the extractive technology of taxol from Taxus baccatal L.. Methods HPLC was applied to determine the content of taxol. The orthogonal test was adopted to examine the effects of the 4 factors consisting of ethanol concentration,ethanol volume,extractive times and extractive time. Results Ethanol concentration and ethanol volume were essential factors influencing the extraction of taxol. High concentration and amount of ethanol facilitated the extraction. Extractive times and extractive time were also important. By single-factorial effect,2 h and 2 times in each extraction of taxol were the best. Conclusion The optimum extractive condition is T9 (A3B3C2D1) and T8 (A3B2C1D3).

AIM: To observe the curative effect of PDT combined with intravitreal injection of ranibizumab treatment for age-related macular degeneration with choroidal neovascularization ( CNV) .
METHODS:In accordance with the inclusion criteria, by indocyanine green choroidalangiography ( ICGA ) and optical coherence tomography ( OCT ) examination confirmed the diagnosis of macular CNV in 27 patients (27 eyes), treated with PDT 3 ~ 7d professional intravitreal injection of ranibizumab. At 1, 3, 6mo after treatment, the results of best corrected visual acuity (BCVA), FFA, ICGA, OCT examination and complications were observed.
RESULTS: The BCVA improved in 17 eyes ( 63%) , stable in 6 eyes ( 22%) , and decreased in 4 eyes ( 15%) . Before treatment, the average leakage area was 1 005. 69±105. 47μm, it were 875. 54 ± 103. 27μm, and 423. 37 ±79.68μm at 1 and 3mo after treatment, there were significant differences compared with before treatment ( P<0. 01). Average central macular thickness of retina before treatment was 485. 58±122. 59μm, and 398. 84±105. 32μm, 297. 74±89. 18μ m at 1 and 3mo after treatment, there were significant differences compared with before treatment( P<0. 01).
CONCLUSION: The method that PDT closed CNV combined with intravitreal injection of ranibizumab can effectively block angiogenesis recurrence, reduce the number of PDT treatment again and complications, improve the therapeutic effect.

To establish the method of HPLC-fingerprint analysis for the quality control of Cnidium monnieri L. Cuss., and identify its active constituents by HPLC-MS, 35 batches of samples were analyzed on a Shimadzu C18 column with a gradient of acetonitrile and 0.1% aqueous aceticacid at a flow rate of 1.0 mL x min(-1) and detected at 245 nm and 322 nm. Furthermore, the typical samples were detected by HPLC-DAD-MS under negative ion mode. 35 batches of Cnidium monnieri L. Cuss. samples were classified into four types based on the results of similarity analysis. According to the comparison of the t(R), MS data and UV maximum absorbance (lambda(max)) values with the standards, 8, 7, 4 and 2 coumarins components were identified in four types of Cnidium monnieri L. Cuss. extracts, separately. The method is repeatable and reliable, and it is capable of effectively controlling the quality of Cnidium monnieri L.
Chromatography, High Pressure Liquid ; methods ; Cnidium ; chemistry ; Coumarins ; analysis ; Ecosystem ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Seeds ; chemistry ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; Spectrophotometry, Ultraviolet

OBJECTIVETo develop an HPLC-UV-ELSD method for the determination of aucubi, harpagide, harpagoside, angoroside C and cinnamic acid in Scrophularia ningpoensis.

METHODThe analytical column was SHIMADZU C18 (4.6 mm x 250 mm, 5 microm). The mobile phase was acetonirile-0.4% acetic acid in a gradient elution. Initial conditions was 5% A; 0-20 min, changed to 10% A; 20-50 min, to 55% A. The flow rate was 0.8 mL x min(-1) and the column temperature was 30 degrees C. The UV detector wavelength was set at 280 nm for harpagoside, angoroside C and cinnamic acid, and the evaporative light-scattering detector (ELSD) drift tube temperature was 105 degrees C, the flow rate of nebulizer gas was 1.2 L x min(-1) for aucubi and harpagide.

RESULTAucubi, harpagide, harpagoside, angoroside C and cinnamic acid was separated well. The linear calibration curves were obtained over of 0.752-13.536 microg for aucubi (r=0.9993, n=6), 0.8280-14.90 microg for harpagide (r=0.9994, n=6), 0.6360-11.45 microg for harpagoside (r=0.9997, n=6), 0.5440- 9.792 microg for angoroside C, (r=0.9997, n=6) and 0.0108-0.1939 microg for cinnamic acid (r=0.9999, n=6). The mean recovers of five compounds were 98.12%, 99.14%, 100.21%, 98.17% and 100.35% with RSD of 2.3%, 1.5%, 1.9%, 1.7% and 0.5%.

CONCLUSIONThis method could simultaneously determinate the content of the five compounds in the S. ningpoensis.

Plant samples were collected and investigated periodically. According to the growth of different parts and the characteristics of dry substance accumulation of Scrophularia ningpoensis, the development of S. ningpoensis could be divided into four stages: seeding stage, stem and leaf growth stage, expanding period of root tubers, and dry substance accumulation stage of root tuber. Leaf numbers of S. ningpoensis grew gradually from one at first to 370 at the final stage, main stem leaf were 50 pieces. Leaf size increasesed with the fastest growth at the stem and leaf growth stage, average daily increase amount was 225 cm2. By the middle of August, leaf size reached to 16,270 cm2. Leaf area indexrose sharply in the seeding stage, and remained above 8 among stem and leaf growth stage and expanding period of root tubers, and rapidly reduced to zero in the stage of dry substance accumulation of root tuber. Leaf area ratio has a tendency of obvious dropping. The net assimilation rate had a small change ranges, two small peak were seeding stage and dry substance accumulation of root tuber. The value of specific leaf area was higher in seeding stage, and in the earlier stage of dry substance accumulation of root tuber. Relative growth rate changed with large ranges, higher in seeding stage, rapid decrease in stem and leaf growth stage, rose in expanding period of root tubers, and declined again in the stage of dry substance accumulation of root tuber. Crop growth rate was higher in the first and last stages, and smaller in interim stage. The growth parameters of S. ningpoensis such as relative growth rate, net assimilation rate, leaf area index, leaf area ratio, specific leaf area, crop growth rate changed along with the growth. The rule of dry matter accumulation was as follows: the dry matter increased slowly during the seeding stage and speeded up in the middle and late stages, and in dry substance accumulation of root tuber increased slower, the growth of dry matter all appeared an "S" curve, and accorded with logistic equation. Cultivation technologies of S. ningpoensis and the relevant management methods could be established according to the growth of different parts of S. ningpoensis and the characteristics of dry substance accumulation in different stage.
China ; Conservation of Natural Resources ; Plant Leaves ; growth & development ; Plant Roots ; growth & development ; Plant Stems ; growth & development ; Plant Tubers ; growth & development ; Scrophularia ; growth & development

The aimed of this study was to prepare stabilized thiomers to overcome the poor stability character of traditional thiomers. Poly(acrylic acid)-cysteine (PAA-Cys) was synthesized by conjugating cysteine with poly(acrylic acid) and poly(acrylic acid)-cysteine-6-mercaptonicotinic acid (PAA-Cys-6MNA, stabilized thiomers) was synthesized by grafting a protecting group 6-mercaptonicotinic acid (6MNA) with PAA-Cys. The free thiol of PAA-Cys was determined by Ellmann's reagent method and the ratio of 6MNA coupled was determined by glutathione reduction method. The study of permeation enhancement and stabilized function was conducted by using Franz diffusion cell method, with fluorescein isothiocyanate dextran (FD4) used as model drug. The influence of polymers on tight junctions of Caco-2 cell monolayer was detected with laser scanning confocal fluorescence microscope. The results indicated that both PAA-Cys and PAA-Cys-6MNA could promote the permeation of FD4 across excised rat intestine, and the permeation function of PAA-Cys-6MNA was not influence by the pH of the storage environment and the oxidation of air after the protecting group 6MNA was grafted. The distribution of tight junction protein of Caco-2 cell monolayer F-actin was influenced after incubation with PAA-Cys and PAA-Cys-6MNA. In conclusion, stabilized thiomers (PAA-Cys-6MNA) maintained the permeation function compared with the traditional thiomers (PAA-Cys) and its stability was improved. The mechanism of the permeation enhancement function of the polymers might be related to their influence on tight junction relating proteins of cells.
Acrylic Resins ; chemistry ; Actins ; metabolism ; Animals ; Caco-2 Cells ; Cysteine ; chemistry ; Dextrans ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Glutathione ; Humans ; Intestinal Absorption ; Intestinal Mucosa ; drug effects ; Nicotinic Acids ; chemistry ; Rats ; Sulfhydryl Compounds ; chemistry

Based on the Bacillus sp., isolated from Lop Nur Desert, the technology of separation and purification and the antioxidant effect were studied. After centrifugation and vacuum filtration, the deproteinization of supernatant was operated with Sevag reagent. The crude exopolysaccharide (EPS) was obtained by precipitation with ethanol. The optimum conditions for the isolation were as follow: pH 7.0, temperature 4?C, time 1.5 h, and material to ethanol ratio 1: 4. Dissolved in water, the crude EPS was fractional separated on activated carbon column (1.5 cm ? 24 cm), eluted with distilled water, 60% ethanol, 95% ethanol, and the main fraction was collected. Then the EPS was purified on Sephadex G-100 gel column, eluted with NaCl (0.2 mol/L). Fractions (4 mL, each) were also combined according to total sugar by phenol-sulfuric acid method and protein content was determined by Coomassie brilliant blue. The results showed that EPS was relatively homogeneous glycoprotein. The data of antioxidation in vitro showed that the EPS had a high antioxidant activity, which could quench hydroxyl radical, superoxide radical and had antilipid peroxidation activity. All of these indicated that EPS was a good natural antioxidant.

Objective To study the exocrine pancreatic function in critically ill children with septic shock,sepsis and hyperlactacidemia.Methods A total of 64 critical pediatric patients were admitted from Jan 2009 to Oct 2012,and clinical and laboratory findings including pancreatic function,and histopathological features and score after autopsy were reviewed.Results (1) Compared with non-septic shock children,the pancreatic pathology score and serum lipase in septic shock group were significantly higher and serum calcium was significantly reduced (P ＜0.05) ; (2) The pancreatic histopathology score was significantly increased in patients with elevated plasma lactate ≥2 times (P ＜0.05),but there were no significant differences in serum calcium and blood amylase and lipase between patients with elevated plasma lactate level and patients with normal plasma lactate level; (3) The concentrations of serum amylase,lipase and urinary amylase were significantly increased in patients with pancreatic histopathology score ＞4 points compared with score ≤4 points patients,but there were no significant differences in above three biomarkers between patients with score ≤3 points and patients with score ＞3 points.Conclusions The pancreas is vulnerable to damage easily occurred in septic shock children especially complicated with hyperlactacidemia.The pancreatic histopathology score ＞ 4 points can be as a sensitive and reliable indicator of pancreas damage.

The whole herb of Solanum nigrum L. can be used as the herbal drug. In this study, UHPLC-Q Exactive high resolution mass combined with GNPS molecular network was used for the rapid characterization of the components in the leaves of S. nigrum L. A total of 157 compounds were identified, including 30 steroid alkaloids, 61 steroid saponins, 35 flavonoids, and 31 other compounds (amino acids and organic acids), by comparison with the data reported in the literature, and mass fragmentation characteristics analysis, as well as the correlation of known and unknown nodes in the GNPS molecular network. Compared with the fruits and stems, the leaves of S. nigrum L was rich in a variety of steroidal saponins, steroidal alkaloids, and flavonoids, and the results lay the foundation for the precise resources utilization of S. nigrum L.