Objective:To optimize the extraction process of the total flavonoids from Ephedrae Radix Et Rhizoma. Methods:The purification method of the total flavonoids from Ephedrae Radix Et Rhizoma was optimized with the yield and content of the total fla-vonoids as the indices. Based on the above research, the process parameters were optimized by an orthogonal test. Results:The opti-mum purification conditions were as follows:the volume fraction of ethanol was 50%, the stirring extraction time was 20 min, and the liquid-solid ratio was 8∶ 1(ml·g-1). Conclusion:The optimum purification technology is simple and reproducible, and suitable for the industrial production.

Objective:To optimize the extraction process for the total saponins from the root of Thladiantha dubia Bunge. Meth-ods:The extraction technology was optimized by orthogonal experiment with the dissolution content of total saponins as the index and the extraction times, extraction duration and ratio of solid to liquid as the influencing factors. Results:The optimum extraction condi-tions for the total saponins from the root of Thladiantha dubia Bunge were as follows:the reflux extraction was conducted twice(1. 5 h per time) with 70% ethanol as the solvent, and the ratio of solid to liquid was 1 ∶6. Conclusion:The optimum extraction technology is simple, reproducible and stable.

Objective:To establish an HPLC-ELSD method for the determination of mahuannin A in ephedrae radix et rhizoma. Methods:The content of mahuannin A was determined by an HPLC-ELSD method on an Alltima TM C18 column (250 mm × 4. 6 mm, 5 μm). The mobile phase was acetonitrile-water (28∶ 72) with a flow rate of 0. 7 ml·min-1, and the column temperature was 30℃. The temperature of drift tube heater was 105℃ and the flow rate of carrier gas was 2. 8 L·min-1 . Results:The linear range of mahua-nnin A was 42. 56-383. 04 μg·ml-1(r=0. 999 8). The average recovery and RSD was 99. 9% and 1. 96%(n=6), respectively. Conclusion:The method is simple and the result is accurate. It can be used for the quality control of ephedrae radix et rhizaoma.

Objective: To establish an HPLC-ELSD method for the content determination of rosolic acid in Radix Rubi.Methods: The ELSD-HPLC content determination of rosolic acid was set up using a Vision HT C 18 HL column(250 mm× 4.6 mm ,5 μm), the mobile phase was methanol-0.1% trifluoroacetic acid with a flow rate of 0.8 ml·min-1 , the column temperature was 35℃, the temperature of drift tube heater was 90 ℃ and the gas was with the flow rate of 2.3 L ·min-1 .Results: The linear range of rosolic acid was 0.155-4.346 μg (r=0.999 9).The average recovery was 100.4% with RSD of 1.88% (n =6).Conclusion: The method is simple and accurate.It can be used for the quality control of Radix Rubi.

Objective:To establish the quality standard for Herba Euphorbia esulae. Methods:A TLC method was used to identify Herba Euphorbia esulae. An HPLC method was used to determine the content of quercetin with the following conditions: an AlltimaTM C18column(150 mm ×4.6 mm,5 μm) was eluted with the mobile phase of acetonitrile-0.4％ phosphoric acid(30 :70), the flow rate was 1. 0 ml·min-1 , the detection wavelength was 360 nm and the column temperature was 35℃. Results:The characteristic spots of samples were the same as those of the standard samples. The linear range of quercetin was 3.194-102.208 μg·ml-1(r=0.9999) and the average recovery was 99. 0％ (RSD=1. 68％, n=6). Conclusion:The method is quick, simple and accurate, which can be used for the quality control of Herba Euphorbia esulae.

Objective To investigate the effects of glucagon-like peptide-1 (GLP-1) on liver oxidative stress, TNF-αand TGF-β1 in rats with diet-induced non-alcoholic fatty liver disease (NAFLD). Methods Thirty male rats were randomly divided into 3 equal groups and fed for 16 weeks with normal diet (ND), high-fat diet (HFD), or high-fat diet with intraperitoneal injection of liraglutide (GLP-1, administered in the later 4 weeks). The rats were then sacrificed to obtain blood samples and liver tissues for analyzing the levels of blood aminotransferase (ALT), triglyceride (TG), and total-cholesterol (TC) using an automatic biochemical analyzer and the levels of superoxide dismutase (SOD), malondial-dehyde (MAD), free fatty acid (FFAs), TNF-αin the liver homogenates and TGF-β1 in serum by radioimmunoassay or ELISA. Results Compared with ND group, HFD group showed significantly increased body weight, liver index, serum levels of ALT, TG, TC, and TGF-β1, and TG, TC, MAD, FFAs, and TNF-a in the liver homogenates, with also significantly increased degree of hepatic steatosis and inflammation activity (P<0.05) and lowered level of SOD. All these changes were markedly ameliorated in GLP-1 group (P<0.05). Conclusion Liraglutide can reduce high-fat diet-induced hepatic steatosis, improve oxidative stress and lipid peroxidation, and decrease TGF-β1 and TNF-a levels in serum and liver homogenates, suggesting its potential as a therapeutic agent for NAFLD.

Objective To investigate the effects of glucagon-like peptide-1 (GLP-1) on liver oxidative stress, TNF-αand TGF-β1 in rats with diet-induced non-alcoholic fatty liver disease (NAFLD). Methods Thirty male rats were randomly divided into 3 equal groups and fed for 16 weeks with normal diet (ND), high-fat diet (HFD), or high-fat diet with intraperitoneal injection of liraglutide (GLP-1, administered in the later 4 weeks). The rats were then sacrificed to obtain blood samples and liver tissues for analyzing the levels of blood aminotransferase (ALT), triglyceride (TG), and total-cholesterol (TC) using an automatic biochemical analyzer and the levels of superoxide dismutase (SOD), malondial-dehyde (MAD), free fatty acid (FFAs), TNF-αin the liver homogenates and TGF-β1 in serum by radioimmunoassay or ELISA. Results Compared with ND group, HFD group showed significantly increased body weight, liver index, serum levels of ALT, TG, TC, and TGF-β1, and TG, TC, MAD, FFAs, and TNF-a in the liver homogenates, with also significantly increased degree of hepatic steatosis and inflammation activity (P<0.05) and lowered level of SOD. All these changes were markedly ameliorated in GLP-1 group (P<0.05). Conclusion Liraglutide can reduce high-fat diet-induced hepatic steatosis, improve oxidative stress and lipid peroxidation, and decrease TGF-β1 and TNF-a levels in serum and liver homogenates, suggesting its potential as a therapeutic agent for NAFLD.

OBJECTIVE: Major depressive disorder (MDD) is a common mood disorder associated with several psychophysiological changes like disturbances of sleep, appetite, or sexual desire, and it affects the patients' life seriously. We aimed to explore a genetic method to investigate the mechanism of MDD. METHODS: The mRNA expression profile (GSE53987) of MDD was downloaded from Gene Expression Omnibus database, including 105 samples of three brain regions in post-mortem tissue suffered from MDD and unaffected controls. Differentially expressed genes (DEGs) in MDD were identified using the Limma package in R. Gene Ontology functions and Kyoto Enrichment of Genes and Genomes pathways of the selected DEGs were enriched using Database for Annotation, Visualization and Integrated Discovery. Protein-protein interactive network of DEGs was constructed using the Cytoscape software. RESULTS: Totally, 241 DEGs in MDD-hip group, 218 DEGs in MDD-pfc group, and 327 DEGs in MDD-str group were identified. Also, different kinds of biological processes of DEGs in each group were enriched. Besides, glycan biosynthesis of DEGs in MDD-str group, RIG-I-like receptor signaling and pyrimidine metabolism of DEGs in the MDD-hip group were enriched, respectively. Moreover, several DEGs like PTK2, TDG and CETN2 in MDD-str group, DCT, AR and GNRHR in MDD-pfc group, and AKT1 and IRAK1 in MDD-hip group were selected from PPI network. CONCLUSION: Our data suggests that the brain striatum tissue may be greatly affected by MDD, and DEGs like PTK2, GALNT2 and GALNT2 in striatum, AR in prefrontal cortex and IRAK1 and IL12A in hippocampus may provide novel therapeutic basis for MDD treatment.
Appetite ; Biological Processes ; Brain ; Depressive Disorder, Major* ; Gene Expression ; Gene Ontology ; Genome ; Hippocampus ; Metabolism ; Microarray Analysis* ; Mood Disorders ; Prefrontal Cortex ; RNA, Messenger*

OBJECTIVETo investigate the effects of glucagon-like peptide-1 (GLP-1) on liver oxidative stress, TNF-α and TGF-β1 in rats with diet-induced non-alcoholic fatty liver disease (NAFLD).

METHODSThirty male rats were randomly divided into 3 equal groups and fed for 16 weeks with normal diet (ND), high-fat diet (HFD), or high-fat diet with intraperitoneal injection of liraglutide (GLP-1, administered in the later 4 weeks). The rats were then sacrificed to obtain blood samples and liver tissues for analyzing the levels of blood aminotransferase (ALT), triglyceride (TG), and total-cholesterol (TC) using an automatic biochemical analyzer and the levels of superoxide dismutase (SOD), malondial-dehyde (MAD), free fatty acid (FFAs), TNF-α in the liver homogenates and TGF-β1 in serum by radioimmunoassay or ELISA.

RESULTSCompared with ND group, HFD group showed significantly increased body weight, liver index, serum levels of ALT, TG, TC, and TGF-β1, and TG, TC, MAD, FFAs, and TNF-α in the liver homogenates, with also significantly increased degree of hepatic steatosis and inflammation activity (P<0.05) and lowered level of SOD. All these changes were markedly ameliorated in GLP-1 group (P<0.05).

CONCLUSIONLiraglutide can reduce high-fat diet-induced hepatic steatosis, improve oxidative stress and lipid peroxidation, and decrease TGF-β1 and TNF-α levels in serum and liver homogenates, suggesting its potential as a therapeutic agent for NAFLD.

Alanine Transaminase ; blood ; Animals ; Cholesterol ; blood ; metabolism ; Fatty Acids, Nonesterified ; metabolism ; Hypoglycemic Agents ; pharmacology ; Liraglutide ; pharmacology ; Liver ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Non-alcoholic Fatty Liver Disease ; blood ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Transforming Growth Factor beta1 ; blood ; Triglycerides ; blood ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective: To express envelope protein of ZIKA virus in baculovirus expression system. Methods: Full-length E gene of ZIKA virus was obtained by DNA synthesis and inserted into vector pFastBac1. The constructed recombinant baculovirus transfer vector pFB1-E was transformed to competent DH10Bac cells. The obtained skeleton plasmid rBacmid-E was transfected to sf9 cells, and the constructed recombinant baculovirus rBac-E was determined for titer, for insertion of E gene by PCR, and for expression of E protein by IFA and Western blotting. Results: PCR proved that skeleton plasmid rBacmid-E was constructed correctly. The titer of rBac-E of passage 3 was 2.58×10⁵pfu/ml. The genome of infected cells virus was extracted, the gene band at length of 3 830 bp was observed after PCR amplification. Indirect immunofluorescence of the infected cells showed the specific green fluorescence, 55×10³specific band was determined by Western blotting identification in the cell pellet of the infected recombinant baculovirus rBac-E. Conclusions The recombinant baculovirus with E gene of ZIKA virus was successfully constructed, which laid a foundation of further study on the function of E protein and the vaccine of ZIKA virus.