Objective To explore whether phenylalanine affect Cdc42, Racl, and RhoA expression and disturb dendritic development. To determine the effects of brain-derived neurotrophic factor (BDNF) on this process. Methods Neurons were cultivated up to 3 days and then treated with 0.9 mmol/L phenylalanine or 100 ng/ml BDNF. Dendritic number were determined by morphologic analysis. Cdc42, Racl, and RhoA protein expression were examined by Western blotting analysis. ResultsThe number of dendrites in cultured neurons reduced two days after being treated with phenylalanine,while BDNF could rescue this change(P < 0.01), furthermore, BDNF was found to inhibit phenylalanineinduced down-regulation of Cdc42, Racl, and RhoA protein expression(P < 0.01). Conclusions Our study indicated that the protective effect of BDNF against phenylalanine-induced neuronal injury is probably mediated by expression of Cdc42, Racl,and RhoA. It suggested a potential neuroprotective action of BDNF in prevention and treatment of brain injury in the patients with phenylketonuria.

OBJECTIVE:To study the stability of Fuke zhidai capsule at accelerated testing and room temperature. METHODS:HPLC was conducted to determine the content of berberine hydrochloride in the preparation:the column was Diamonsil C18 with mobile phase of B methanol-0.7% triethylamine(pH adjusted to 3 by phosphate)(50:50,V/V)at a flow rate of 1 ml/min;detec-tion wavelength was 265 nm,with a column temperature at 45 ℃,and the injection volume was 5 μl;its moisture,disintegration time,accelerated stability and long-term stability were studied according to the 2015 edition of Chinese Pharmacopoeia. RE-SULTS:The linear range of berberine hydrochloride was 0.0217-0.1736 mg/ml(r=0.9999);RSDs of precision,stability and re-producibility tests were lower than 2%;recovery was 97.24%-101.73%(RSD=1.72%,n=6). CONCLUSIONS:The moisture,dis-integration time,accelerated stability and long-term stability were all in line with the quality standards.

AIM:To select an optiomal formulation of liposome entrapped alkanin prepared with ethanol injection in the combination of orthogonal design and central composite design/Response surface method.METHODS:Five influencing factors,including ratio of alkanin to liposome and soyabean lecithin to cholesterol,volume ratio of ethanol phase to water phase,soybean concentration and mannitol concentration were chosen as indices by orthogonal design.Then the method utilized the central composite design,which fitted multivariate linear and second/third order nonlinear regression equation,to find out the optimal formulation.RESULTS:Third order regression equation gained made the good prediction about entrapment(85.4%) and alkanin load(5.16%),and exhibited no significant difference between theoretic value and measured value.CONCLUSION:The combination of orthogonal design with central composite design is a feasible and convenient method to optimize the prescription of alkanin-lipo-some.

AIM:To select an optiomal formulation of liposome entrapped alkanin prepared with ethanol injection in the combination of orthogonal design and central composite design/Response surface method.METHODS:Five influencing factors,including ratio of alkanin to liposome and soyabean lecithin to cholesterol,volume ratio of ethanol phase to water phase,soybean concentration and mannitol concentration were chosen as indices by orthogonal design.Then the method utilized the central composite design,which fitted multivariate linear and second/third order nonlinear regression equation,to find out the optimal formulation.RESULTS:Third order regression equation gained made the good prediction about entrapment(85.4%)and alkanin load(5.16%),and exhibited no significant difference between theoretic value and measured value.CONCLUSION:The combination of orthogonal design with central composite design is a feasible and convenient method to optimize the prescription of alkanin-liposome.

The purpose of the study is to use O. intermedium DN2 to degrade nicotine in tobacco extracts for making reconstituted tobacco. Firstly, we studied the effects of various factors on degradation of nicotine in the extracts by strain DN2. When we added 0.1% yeast extract into the extracts, adjusted its pH value to 7.0 by ammonia solution, inoculated 15% cultures and maintained fermentation temperature of 30 degrees C, the degradation rate of nicotine by strain DN2 was the fastest. Furthmore, under these conditions, we studied the degradation rates of nicotine in three fed batches culture which carried out in a 30-L reactor, the result showed that the average degradation rate of nicotine by strain DN2 was 140.55 mg/L/h, which was much higher than that reported in other studies. These results indicated that strain DN2 may be useful for reducing nicotine content of reconstituted tobacco.
Nicotine ; metabolism ; Ochrobactrum ; classification ; metabolism ; Plant Extracts ; metabolism ; Tobacco ; chemistry

Aims To establish a UPLC-MS method for quantifying protocatechuie acid,kaempferol-3-O-β-D-glucoside and quercitrin and to investigate the pharma-cokinetics of three indix components in flower of Poly-gonumorientale L.in rat plasma.Methods The anal-ysis was achieved by BEH C18 column (2.1 mm ×100 mm,1.7 μm)with a mobile phase composed of 0.1%formic acid using step gradient elution.A TQD tandem mass spectrometry equipped with electrospray ionization source was used as detector and operated by selected ion recording(SIR)mode.Results In the selected linear range,calibration curves of the three markers components showed good linearity.Extraction recovery rate,precision,accuracy and stability reached the de-termination request.The parameters of Tmax (h ) in three index components were 0.46 ±0.1,0.79 ±0.33 and 2.63 ±4.6,respectively;Cmax (μg·L -1 )in three index components were 463.8 ±207.81,18.53 ±7.82 and 137.38 ±71.09,respectively.Conclusion The fully validated UPLC-MS method has been successfully applied to the pharmacokinetic study of the three index components in flower of Polygonumorientale L.in rat plasma.

Objective To investigate the specific expression of endoplasmic reticulum stress-related GRP78 and CHOP in podocyte of MKR mice with diabetic nephropathy (DN);To discuss the intervention effect of Zuogui Jiangtang Yishen Decoction (ZGJTYS). Methods 8-week old MKR mice were randomly divided into five groups:blank group, model group, ZGJTYS group, 4-Phenylbutyric acid group, gliquidone+benazepril group, and normal control group consisting of wild type C57BC/6 mice, 10 mice per group. All mice from model group and each treatment group received high-fat diet feed and unilateral nephrectomy to make the DN model. Mice from treatment groups received medicine intervention for four weeks. The levels of FBG were detected by electrochemical detection method, and the UmAlb was detected by ELISA. The expressions of GRP78, CHOP mRNA, and protein were detected by RT-PCR and Western blot. The morphological structure changes of the podocytes were observed by transmission electron microscopes. Results Compared with blank group, FBG and UmAlb in the model group increased significantly (P<0.01);the expressions of GRP78 mRNA and protein markedly decreased (P<0.01);the expressions of CHOP mRNA and protein increased (P<0.01). Compared with the model group, FBG and UmAlb in each treatment group significantly decreased (P<0.01);the expressions of GRP78 mRNA and protein in the ZGJTYS group and 4-Phenylbutyric acid group increased (P<0.01), the expressions of CHOP mRNA and protein decreased (P<0.01). The renal pathological lesions of each treatment group were significantly improved compared with model group. Conclusion ZGJTYS Decoction can effectively improve the damaged podocyte induced by endoplasmic reticulum stress, delay DN progress.

Objective To evaluate the effects of isokinetic quadriceps training with different ranges of motion for improving mal-patellar tracking after total knee replacement (TKR). Methods Isokinetic quadriceps training was administered to 52 TKR cases, involving 66 knee joints with poor patellar tracking. The subjects were randomly allocated into 3 groups for isokinetic training with ranges of 0-30°, 30-60° or 60-90°, with 22 knee joints in each group. The angular velocity was set at 30°/second. X-rays with loading and with 45° knee flexion were taken before and after the training. The lateral patello-femoral angle,the patella index and the patello-femoral index were recorded along with the maximum torque ( MT), the total work ( TW ), and the average work(AW) in the three different ranges of motion. Hospital for Special Surgery (HSS) scores were also recorded. Results After 3 months of training, the average lateral patello-femoral angle, patella index, and patello-femoral index in the 0-30° training group were all significantly better than before the training. However, there was no significant difference in these measures for the 30-60° and 60-90° groups. After training average MT, TW and AW increased significantly in all groups, but the improvements were significantly correlated with the differences in lateral patello-femoral angles, patella indices and patello-femoral indices only for the 0-30°group. When compared with the baseline values, a statistically significant difference in HSS score after training was observed only in the 0-30° training group. Conclusions Isokinetic quadriceps training with a 0-30° range of motion can best realign patellar tracking after TKR, and best improve knee function.

Aim To evaluate the inhibitive and induc-tive effects of Polygonum capitatum water extract on main cytochrome P450 isoforms in human and liver mi-crosomes of mouse in vitro for predicting the herb-drug interactions in clinical application. Methods The in vitro inhibitory effect was evaluated by incubating Po-lygonum capitatum water extract with the probe sub-strates of main phase I metabolic enzymes in human liver microsomes, including CYP1A2, CYP2E1, CYP2C9,CYP2C19 and CYP3A4. Mice were adminis-tered with Polygonum capitatum water extract at dosage of 0 . 58 g · kg-1 and 1 . 16 g · kg-1 by gastric lavage for successive 7 days and 14 days, then the cocktail-LC-MS/MS method was applied to assess the inductive effect of main CYP450 isoforms in mouse liver micro-somes. Results The IC50 values of Polygonum capita-tum water extract on main CYP450 isoforms in human liver microsomes were from 849 . 6 mg · L-1 to 2 287 mg·L-1 . Compared with the blank control group, the activites of CYP2C9 and CYP3A4 in 1. 16 g·kg-1 7 d group were about 49 . 9 % and 21. 1 % higher ( P <0. 01, P < 0. 05 ) respectively, the activities of CYP2C9 and CYP3A4 in 0. 58 g·kg-1 7 d group were 27. 6 % and 15. 5 % higher ( P <0. 01 , P <0. 05 ) respectively, the activities of CYP2C9 and CYP3A4 in 1. 16 g·kg-1 14 d group were 67. 5 % and 32. 1 %higher (P<0. 01) respectively, while the activities of CYP1 A2 , CYP2 E1 and CYP2 C19 were not increased significantly in Polygonum capitatum treatment group. Conclusions Polygonum capitatum water extract do not show the inhibitory effect on main CYP450 in hu-man liver microsomes. There is induction on CYP2C9 and CYP3 A4 in mouse liver microsomes by Polygonum capitatum water extract.

To study the comparability of myocardial enzyme spectrum detection results by two different bio-chemical testing systems to provide references for mutual accredit of detection results in one lab. Following NC-CLS document EP9-A2, 8 clinical specimens were selected every day for five days and tested for AST, LDH, CK and CK-MB respectively with Siemens ADVIA1800 and Johnson & Johnson Victro350 systems. Acknowledging Siemens AD-VIA1800 as objective testing system, the results were utilized to make correlation analysis and clinical acceptability esti-mation. There was high precision in Siemens ADVIA1800 and Johnson & Johnson Victro350 (CV≤1/3CLIA′88 allowable error). It's proved by t test that the difference of the myocardial enzyme results tested with Siemens ADVIA1800 and Johnson Victro350 was statistically significant (P<0.01), while the two testing systems showed significant correlation (r>0.975). In the comparison with the Siemens ADVIA1800, the standard errors (SE%) of AST, CK and CK-MB tested with Johnson Victro350 were 4.0%-6.6%, 9.6% and 13.3%, which were accepted by clinical standard; while the SE% of LDH tested with Johnson Victro350 were 11.2%-12.7%, which were beyond clinically acceptable range. In case one item is tested by different testing systems in one laboratory, the comparison and bias estimation acceptable eval-uationare necessary to judge the clinical acceptability, which can ensure the accuracy and consistency of the results.