Aim To determine potential biomarkers contributed to occurrence, development and recovery of ethanol-induced liver injury in rat and elucidate hepatoprotective effect of Yin Chen Hao Tang based on metabonomic investigation. Methods A UPLC-Q-TOF/MS based metabonomic method was developed for investigating trajectory change and inter-relationship of urinary metabolome of rats with different treatments. Results Four potential biomarkers were determined which contributed to occurrence, development and recovery of ethanol-induced liver injury in rat, and Yin Chen Hao Tang could significantly recover trajectory change in disorder. Conclusion The developed method was successfully applied to investigate ethanol-induced liver injury in rat, and also hepatoprotective effect of Yin Chen Hao Tang was elucidated.

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; chemistry ; genetics ; growth & development ; metabolism ; Poultry Diseases ; virology ; Protein Structure, Tertiary ; Spike Glycoprotein, Coronavirus ; chemistry ; genetics ; metabolism ; Transfection ; Vero Cells

In order to evaluate the characteristics of the spray drying of total flavonoids of Epimedium extracts assisted with soybean polysaccharide, a certain percentage of soybean polysaccharide or polyvidone were added to the total flavonoids of Epimedium extract to conduct the spray drying. The effect of soybean polysaccharides against the wall sticking effect of the spray drying was detected, as well as the powder property of total flavonoids of Epimedium spray drying powder and the dissolution in vitro behavior of the effective component. Compared with the total flavonoids of Epimedium spray drying powder, soybean polysaccharide revealed a significant anti-wall sticking effect. The spray drying power which had no notable change in the grain size made a increase in the fluidity, improvement in the moisture absorption and remarkable rise in the dissolution in vitro behavior. It was worth further studying the application of soybean polysaccharide in spray drying power of traditional Chinese medicine.
Epimedium ; chemistry ; Flavonoids ; analysis ; Particle Size ; Polysaccharides ; chemistry ; Powders ; Soybeans ; chemistry

AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

The cholesterol oxidase producing strain Brevibacterium sp.DGCDC-82 was treated with NTG (1 mg/mL)under ultrasonicztion(200 W,50 kHz).A red mutant named Brevibacterium sp.DGCCN-25 showed higher and stable production of cholesterol oxidase was obtained.The enzyme activity was increased by 140%,it is 1.24 U/mL.Then dealed with DGCCN-25 using the same method,two revertants were obtained,one was white and the other was rose pink.The enzyme activity of two revertants was obvious decrease,they are 0.17 U/mL and 0.69 U/mL.The results showed the positive correlation between COD acticity and red pigment producing by Brevibacterium sp..The relativity model can be used as a method of screening for mutation and directed evolution.

OBJECTIVETo study the influecnce of gentian leaf blight on the output and quality of rough gentian.

METHODThe same grade seedlings were transplanted, disease of every plant was investigated in autumn and the output of gentian was determined. HPLC was applied to determine the content of gentiopicroside and swertiamarin.

RESULTThe output decreased with the aggravation of the disease, and the decrease was obvious when the index of disease was above 60. The content of gentiopicroside and swertiamarin began to drop when the index of disease was above 70.

CONCLUSIONThe loss of output and the drop of quality are relatively heavy when the disease is serious. The loss of income is not obvious when the index of disease is under 60.

Gentiana ; chemistry ; growth & development ; Glucosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Mitosporic Fungi ; growth & development ; Plant Diseases ; economics ; microbiology ; Plant Leaves ; microbiology ; Plants, Medicinal ; chemistry ; growth & development ; Pyrans ; analysis ; Pyrones ; analysis ; Quality Control

The mechanism of biological actions of quercetin was studied by using metabolomic method and biomolecular network. HPLC-MS was used to analyze the serum metabolome in rats of blank group and quercetin administration group rats, and MS data were processed by MATLAB software. With multivariate statistical analysis of serum metabolite profiles, a clear separation among blank group and quercetin administration group was achieved, potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval. Four compounds, related enzymes, action targets and metabolic pathways had been confirmed, namely retinoic acid and RARbeta, arachidonate and COX-2, 3, 5-diodotyrosine and TPO, uridine diphosphate glucose and PDEs. The mechanism of quercetin enhancing ability of retinoic acid on the induction of RARbeta, activating TPO, using as COX-2 and PDEs inhibitor was approved by biomolecular network and related literatures. In this study, a mechanism of multiple biological actions of quercetin was evaluated at the level of the biomolecular network, metabolomics and biomolecular network can be used to investigate the biological effects mechanism of quercetin, which provided a new method to further revealing mechanism of drug action.
Animals ; Antioxidants ; pharmacology ; Biomarkers ; blood ; Chromatography, Liquid ; Mass Spectrometry ; Metabolic Networks and Pathways ; Metabolome ; Metabolomics ; Multivariate Analysis ; Quercetin ; pharmacology ; Rats

Objective: To examine and analyze the nm23, p16 gene expression in gastric cancer tissure,and follow up patients 5 years to discuss the relationship between nm23 and p16 gene synergy expression and gastric cancer biological characteristic and prognosis.Methods: nm23 and p16 protein in gastric cancer tissue and control were detected by immunohistochemistry,and the patients had been followed up for 5 years. Results: In 84 samples of gastric cancer, nm23 positive rate was 46.43%, p16 was 44.05%, the positive rate of gastric cancer tissue and metastasitic lymph node was lower than that of normal control, normal tissue near cancer or benign polyp,and these two genes were related to the depth of tumor invasion and clinical stage.The mortality and recurrence-metastasis rate was higher in these low expression group, and had a shorter median survive period. Conclusion: Abnormal expression of nm23 and p16 gene plays a important role in gastric cancer recurrence and devolepment and may be one of markers for evaluating tumor biological behavior and prognosis.

OBJECTIVETo optimize the prescription dose of Mahuang decoction in a multi-target manner, in order to provide reference for the quantitative optimization of the prescription dose of the traditional Chinese medicine compound.

METHODThe number of diaphoretic spots in rats, the tracheal antispasmodic rate in guinea pigs and the writhing times by acetic acid in mice were taken as the indexes for evaluating the diaphoretic, antispasmodic and analgesic effects. According to the experimental results of the 16 orthogonal combination prescriptions, a mathematical dose-effect model was built by support vector regression (SVR) and quadratic response surface regression (RSR) respectively. The multi-target optimization was achieved by elitist non-dominated sorting genetic algorithm (NSGA-II) and entropy weight TOPSIS method.

RESULTThe optimal dose of Mahuang decoction after being optimized by SVR modeling contained 17.71 g of Ephedrae Herba, 9.57 g of Cinnamomi Ramulus, 11.75 g of Armeniacae Semen Amarum and 4.39 g of Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle. The optimized result by RSR modeling contained 13.37 g of Ephedrae Herba, 11.61 g of Cinnamomi Ramulus, 11.98 g of Armeniacae Semen Amarum and 5.67 g of Glycyrrhizae Radix et Rhizoma Praeparate Cum Melle. SVR was superior to RSR in both of the forecast capacity and optimization results.

CONCLUSIONSVR-NSGA-II-TOPSIS method could be adopted for the multi-target optimization for the dose of Mahuang decoction and other traditional Chinese medicine compounds. It is proved to be the optimal prescription with the best efficacy, and could provide scientific quantitative basis for determining the dose of traditional Chinese medicine compound prescriptions and developing new traditional Chinese medicines.

Animals ; Cinnamomum ; chemistry ; Drug Compounding ; methods ; Drug Dosage Calculations ; Drug Prescriptions ; Ephedra ; chemistry ; Ephedra sinica ; chemistry ; Glycyrrhiza ; chemistry ; Guinea Pigs ; Mice ; Rats

Objective To investigate the effects of cognitive interventions (CIs) in the context of communi- ty based rehabilitation (CBR) on cognitive deficits (CDs) in stroke patients.Methods Ninety-two stoke patients with CDs were randomly divided into a CI group and a control group.All patients were treated with conventional CBR.In addition,the patients in the CI group were also treated with special intervention therapy.The patients in both groups were assessed with the neurological and cognitive status examination (NCSE) for cognitive functioning, the FCA for motor function and the BI for their ability in the activities of daily living.Results The NCSE,FCA and BI scores in the cognitive intervention group after treatment were significantly higher than those before treatment and also significantly higher than those in the control group after treatment.Conclusion CIs can not only improve CDs,but also enhance recovery of motor function and ADL.