1.Metabolomic study on preventive effect of Aqueous extract and Volatile oil of Dalbergia Odorifera on myocardial ischemia/reperfusion injury in rats
Fei MU ; Jialin DUAN ; Haixu BIAN ; Rui LIN ; Peijin SHANG ; Zhihui ZHU ; Aidong WEN ; Miaomiao XI
Chinese Pharmacological Bulletin 2016;32(10):1377-1382
Aim To investigate the changes of serum metabolism after the treatment of DOA and DOO on myocardial ischemia reperfusion ( MI/R ) injury in rats, and to explore the pathogenesis of MI/R injury and drug action mechanism. Method The serum samples of Sham group, Model group, DOA group and DOO group of rats were acquired, gas phase time of flight mass spectrometry ( GC-TOF-MS) was applied to analyze the metabolic profiles of the samples. After da-ta preprocessing, they were processed into SIMCA 14. 1 software for multivariate statistical analysis. Results By principal components analysis ( PCA ) , partial least squares analysis ( PLS-DA) and orthogonal partial least squares analysis ( OPLS-DA ) , the Model group and Sham group were obviously separated, the drug in-tervention group and Model group were separated and close to Sham group. The therapeutic effect of DOO and DOA on MI/R injury in rats was proved. The ex-perimental results identified 13 endogenous biomark-ers, which were related to the glucose metabolism,lipid metabolism and amino acid metabolism pathway. Con-clusion DOA and DOO may protect the MI/R injured rats by regulating the glucose metabolism, lipid metab-olism and amino acid metabolism pathway.
2.Related research on a novel macropores calcium phosphate cement as bone tissue engineering scaffold
Tao LIU ; Jian LI ; Kang LU ; Zhihui BIAN ; Jianfei PANG ; Chao YANG ; Zengwu SHAO
International Journal of Biomedical Engineering 2012;35(2):116-120
ObjectiveTo investigate the cell toxicity of a novel macropores calcium phosphate cement (CPC) scaffold and its influence on cell adhesion,growth and proliferation.MethodsA novel CPC material was synthesized by means of adding mannitol porogens and applying sodium solution as the cement liquid.The cell growth and proliferation in the novel CPC material extraction was observed by CCK8 assay.Scanning electron microscopy was used to observe hole diameter of the material,cell adhesion and growth in the material.The experiment of three point bending was used to test the biomechanic performance of the CPC material.ResultsThe novel CPC material reached hole diameter value of (267.43±118.01)μm,microporosity of (66.15±6.91)%.Maximum load,flexural strength and toughness of the novel CPC material was increased about one time compared to the traditional CPC(P<0.05).CCK8 assay showed there were no significant difference of the light absorption value of cells in the CPC extraction in the 4th,6th,8th day compare to the control group (P>0.05).ConclusionThe novel CPC material has the strong biomechanics performance,macropores,high microporosity and excellent biocompatibility,which is promising for ideal bone tissue engineering scaffold.
3.Simultaneous determination of L-tryptophan and L-kynurenine in rat plasma by LC-MS/MS
Kaicheng XU ; Ruilian YU ; Yunfei YANG ; Zhihui BIAN ; Jiawei SHAO
Journal of China Pharmaceutical University 2016;47(6):714-718
To establish a simple and sensitive LC-MS/MS method for simultaneous determination of the concentration for L-tryptophan(L-Try)and L-kynurenine(L-Kyn)in rat plasma. The changesin the process of the liver tumors formation may provide a basis for the diagnosis of liver cancer. 3-Nitro-L-tyrosine(3-NT)were added as the internal standard for the determination of two active substances and the chromatographic analysis was performed on a RRHD Eclipse Plus C18 column(3. 0 mm×100 mm, 1. 8 μm). The mobile phase was composed of water and acetonitrile(containing 0. 1%formic acid)(90 ∶10)at a flow rate of 0. 25 mL/min, and the injection volume is 5 μL. Detection and quantification were performed by mass spectrometry in multiple reaction monitoring mode with m/z 205. 12→146. 10(L-Try), m/z 209. 09 →146. 10(L-Kyn), m/z 227. 09→181. 10(3-NT), respectively. The results show that thelinear ranges were 9. 670-9 670 ng/mL for L-Try, and 9. 973-9973 ng/mL for L-Kyn(r2≥0. 9990). The limit of quantitation were 9. 670 ng/mL for L-Try, and 9. 973 ng/mL for L-Kyn, respectively. The intra- and inter-day precisions were all less than15%; the recoveries ofthe two analytes were more than 81. 17% and severe matrix effect was not observed. The ratio of L-Try/L-Kyn determined by LC-MS/MS in rat plasma showed an overall downward trend, which could used effectively for the drug metabolism studies and researches on the action mechanism of medicine on liver cancer. A rapid, simple, sensitive and specific LC-MS/MS method has been successfully developed and could also be used effectively for the drug metabolism studies and researches on the action mechanism of medicine on liver cancer.
4.Comparative Study on the Contents of Fatty Acids in Isaria cicadae, Cordyceps militaris and Cordyceps sinensis
Zhihui BIAN ; Ruilian YU ; Simin WEI ; Wen JIANG ; Jiawei SHAO ; Zhen YU ; Jinguo XU ; Gaohong LYU
China Pharmacy 2017;28(30):4252-4256
OBJECTIVE:To establish a method for simultaneous determination of fatty acids in Isaria cicadae,Cordyceps militaris and C.sinensis,and to compare the difference of the contents of fatty acids among above medicinal herbs.METHODS:GC-MS method was adopted.Chromatographic condition:the determination was performed on TG-5MS gas phase capillary column with carrier gas of nitrogen at the flow rate of 1.2 mL/min.The inlet temperature was 290 ℃ by splitlesssampling;valve opening time was 1 min,and volume of sample was 1 μL.Mass spectrometry condition:the ion source is an electrospray ionization source.The temperatures of ion source and transmission line were 280 ℃,the initial temperature of the chromatographic column was 80 ℃ (gradient elution),ionization voltage was 70 eV.Solvent delay time was 5 min,and scan mass range was m/z 30-400.RESULTS:The linear ranges of tetradecanoic acid,pentadecanoic acid,palmitic acid,palmitoleic acid,heptadecanoic acid,heptadecenoic acid,docosahexaenoic acid,methyl oleate,linoleic acid,arachidonic acid,eicosenoic acid,diolefinic acid,eicosatrienoic acid,heneicosanoic acid,behenic acid,tricosanoic acid,lignoceric acid were 1.400-44.520 μg/mL(r=0.999 8),2.091-93.721 μg/mL(r=0.999 7),3.146-85.856 μg/mL(r=0.998 2),1.664-61.444 g/rnL(r=0.998 7),1.773-64.983 g/mL(r=0.999 5),1.781-68.421 μg/ mL (r=0.999 7),1.706-55.606 μg/mL (r=0.999 8),1.439-47.989 μg/mL (r=0.999 6),1.738-66.908 μg/mL (r=0.999 6),2.086-94.206 μg/mL(r=0.999 5),1.356-44.966 μg/mL(r=0.999 4),1.444-56.814 μg/mL(r=0.999 7),1.375-52.335 μg/mL(r =0.999 8),1.512-60.312 μg/mL(r=0.999 5),1.450-59.760 μg/mL(r=0.999 7),1.427-58.757 μg/mL(r=0.999 1),1.269-58.109 μg/ mL(r=0.999 3),respectively.The limit of quantitation was no more than 1 764.71 μg/mL,and the limit of detection was no more than 529.42 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0%.The recoveries were 84.87%-108.93% (RSD ranged 0.19%-2.23%,n=6).There were 17 fatty acids in C.militaris,16 fatty acids in Ⅰ.cicadae and 16 fatty acids in C.sinensis.The contents of unsaturated fatty acids in above medicinal herbs were higher than that of saturated fatty acids.The content of fatty acids in artificial cultivated Ⅰ.cicadae was mostly higher than other medicinal herbs.COCLUSIONS:The method is simple,accurate,stable and reproducible.It can be used for simultaneous determination of fatty acids in I.cicadae,C.militaris and C.sinensis.Above 3 kinds of medicinal materials.From the perspective of fatty acid content,the quality of artificial cultivated I.cicadae is best.
5.MicroRNA-638 inhibits cell proliferation by targeting phospholipase D1 in human gastric carcinoma.
Jiwei ZHANG ; Zehua BIAN ; Jialiang ZHOU ; Mingxu SONG ; Zhihui LIU ; Yuyang FENG ; Li ZHE ; Binbin ZHANG ; Yuan YIN ; Zhaohui HUANG
Protein & Cell 2015;6(9):680-688
MicroRNAs (miRNAs) are a type of small non-coding RNAs that are often play important roles in carcinogenesis, but the carcinogenic mechanism of miRNAs is still unclear. This study will investigate the function and the mechanism of miR-638 in carcinoma (GC). The expression of miR-638 in GC and the DNA copy number of miR-638 were detected by real-time PCR. The effect of miR-638 on cell proliferation was measured by counting kit-8 assay. Different assays, including bioinformatics algorithms (TargetScan and miRanda), luciferase report assay and Western blotting, were used to identify the target gene of miR-638 in GC. The expression of miR-638 target gene in clinical CRC tissues was also validated by immunohistochemical assay. From this research, we found that miR-638 was downregulated in GC tissues compared with corresponding noncancerous tissues (NCTs), and the DNA copy number of miR-638 was lower in GC than NCTs, which may induce the corresponding downregulation of miR-638 in GC. Ectopic expression of miR-638 inhibited GC cell growth in vitro. Subsequently, we identified that PLD1 is the target gene of miR-638 in GC, and silencing PLD1 expression phenocopied the inhibitory effect of miR-638 on GC cell proliferation. Furthermore, we observed that PLD1 was overexpressed in GC tissues, and high expression of PLD1 in GC predicted poor overall survival. In summary, we revealed that miR-638 functions as a tumor suppressor in GC through inhibiting PLD1.
3' Untranslated Regions
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genetics
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Apoptosis
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genetics
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Base Sequence
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Cell Line, Tumor
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Cell Proliferation
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genetics
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Down-Regulation
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genetics
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Humans
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MicroRNAs
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genetics
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Phospholipase D
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genetics
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Prognosis
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Stomach Neoplasms
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diagnosis
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enzymology
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genetics
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pathology