1.Optimization of Extraction Technology for the Total Saponins from the Root of Thladiantha Dubia Bunge
Dongyan CHENG ; Xin CHEN ; Lishu WANG ; Jun GAO
China Pharmacist 2016;19(9):1660-1662
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.
2.Determination of Mahuannin A in Ephedrae Radix et Rhizoma by HPLC-ELSD
Dongyan CHENG ; Lishu WANG ; Jun GAO ; Donghong CHENG ; Chaonan WANG
China Pharmacist 2016;19(2):360-361
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.
3.Extraction Technique of Total Flavonoids from Ephedrae Radix Et Rhizoma
Dongyan CHENG ; Lishu WANG ; Jun GAO ; Donghong CHENG ; Chaonan WANG
China Pharmacist 2016;19(3):427-429
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.
4.Study on Quality Standard for Herba Euphorbia Esulae
Lishu WANG ; Dongyan CHENG ; Xin CHEN ; Jun GAO ; Donghong CHENG ; Chaonan WANG
China Pharmacist 2017;20(9):1649-1651
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.
5.Determination of Rosolic Acid in Radix Rubi by HPLC-ELSD
Dongyan CHENG ; Lishu WANG ; Xin CHEN ; Jun GAO ; Donghong CHENG ; Chaonan WANG
China Pharmacist 2017;20(8):1469-1470
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.
6.Microarray Analysis of the Major Depressive Disorder mRNA Profile Data.
Lishu GAO ; Yue GAO ; Enping XU ; Jian XIE
Psychiatry Investigation 2015;12(3):388-396
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
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Biological Processes
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Brain
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Depressive Disorder, Major*
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Gene Expression
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Gene Ontology
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Genome
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Hippocampus
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Metabolism
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Microarray Analysis*
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Mood Disorders
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Prefrontal Cortex
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RNA, Messenger*
7.Effects of glucagon-like peptide-1 on liver oxidative stress, TNF-α and TGF-β1 in rats with non-alcoholic fatty liver disease.
Huiting GAO ; Lishu XU ; Dongfeng LI ; Lichang GUANG ; Weiping DENG
Journal of Southern Medical University 2013;33(11):1661-1664
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
8.New Quality Standard for Schisandrae Chinensis Fructus
Xiaojie LIU ; Dongyan CHENG ; Lishu WANG ; Jun GAO ; Yongxiu ZHANG ; Weimin ZHAO ; Jiasheng XU
China Pharmacist 2018;21(10):1755-1758
Objective: To establish a new standard for the quality evaluation of Schisandrae chinensis Fructus. Methods: On the basis of the 2015 edition pharmacopoeia standard of Schisandrae chinensis Fructus, some testing items including the characteristic chro-matogram (HPLC) and total lignans content (UV) were established. Results: With schizandrin as the reference material, the HPLC specific chromatograms of Schisandrae chinensis Fructus including 7 characteristic peaks were established. The content of total lignans in Schisandrae chinensis Fructus was not less than 1. 8% with schizandrin as the standard. Conclusion: The proposed quality evalua-tion standard is more comprehensive and reproducible. It can provide a more scientific evaluation tool for the quality evaluation of Schisandrae chinensis Fructus and its related preparations.
9. Expression of envelope protein of Zika virus in baculovirus expression system
Hanchun GAO ; Lihong YAO ; Chao WANG ; Lishu ZHENG
Chinese Journal of Experimental and Clinical Virology 2017;31(6):562-565
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.
10.Correlation of BUA, SOS, T-score and SI with bone mineral density in elderly women with knee osteoarthritis and their diagnostic efficacy in osteoporosis
Lishu GAO ; Ri LIU ; Shifeng DAI ; Jun YU ; Kunnan XIE
Journal of Chinese Physician 2023;25(4):537-540
Objective:To analyze the broadband ultrasound attenuation (BUA), speed of sound (SOS), standard deviation of bone density (T-Score) and stiffness index (SI) with bone mineral density (BMD) in elderly female patients with knee osteoarthritis and their diagnostic efficacy of osteoporosis.Methods:Fifty elderly female patients with knee osteoarthritis admitted o Tangshan People′s Hospital from January 2021 to January 2022 were selected as the observation group, and 40 healthy female patients during the same period were selected as the control group. The results of BUA, SI, T-score and SOS in observation group and control group were analyzed and compared. The BUA, SI, T-score and SOS in elderly women with knee osteoarthritis at different ages and with different bone densities were compared, and the diagnostic value of BUA, SI, T-score and SOS in osteoporosis was analyzed by receiver operating characteristic (ROC) curve.Results:The BUA, SI and T-score of observation group were lower than that in the control group, while SOS was higher than that in the control group (all P<0.05). Among elderly female patients with knee osteoarthritis of different ages, the older the age, the lower the BUA, SI and T-score (all P<0.05), while there was no statistical significance in SOS of elderly female patients with knee osteoarthritis of different ages ( P>0.05). In elderly women with knee osteoarthritis with different BMD grades, the BUA, SI, and T-score in the osteoporosis group were lower than those in the osteopenia group and the normal bone group, and the BUA, SI, and T-score in the osteopenia group were lower than those in the normal bone group; the SOS in the osteoporosis group was higher than those in the osteopenia group and the normal bone group, and the SOS in the osteopenia group was higher than those in the normal bone group (all P<0.05). BUA, SOS, T-score and SI had high sensitivity and specificity in the diagnosis of osteoporosis in elderly women with knee osteoarthritis (all P<0.05). Conclusions:BMD in elderly women with knee osteoarthritis is associated with BUA, SI, T-score, and SOS, and has high diagnostic value for osteoporosis.