1.Analysis of Microstructure of Plant Fibers with Different Hemicellulose Content by Solid State Nuclear Magnetic Resonance and Atomic Force Microscope
Jinquan WAN ; Qing XIAO ; Yan WANG
Chinese Journal of Analytical Chemistry 2010;38(3):347-351
Fibers with different hemicellulose contents were produced using various degree removal of hemicellulose to obtain large differences in cellulose and hemicellulose proportions at a similar lignin content. Solid state cross polarisation magic angle spinning carbon-13 nuclear magnetic resonance(CP/MAS ~(13)C-NMR) and atomic force microscope(AFM) had been employed to investigate the microstructure of fibers. The results showed that there was an increase in relative content of para-crystalline cellulose with the decreas of hemicellulose content obtained by the spectral fitting for the cellulose C1-region(δ 102-108). The elementary fibril size was relatively constant between 4.0 and 4.3 nm for the three samples obtained by the spectral fitting for the cellulose C4-region(δ 80-92). The difference in elementary fibril size between the samples was not significant. However, the elementary fibril aggregate size increased from 17.9 to 22.2 nm with the decreas of hemicellulose content, which was a significant change. The results of AFM analysis showed that the fiber with a high hemicellulose content had a porous surface structure. In fibres with a low hemicellulose content, the elementary fibril aggregates formed a much more compact surface structure. Lower hemicellulose content can promote the partially irreversible microfibril aggregation, which caused tensions in the microfibrils due to the finite dimensions of the cell wall, amounting to stress in the microfibrils. However, the porous structure can be improved as hemicellulose content decreased to a certain extent.
3.Effects of road transport on hematological and biochemical parameters in new zealand rabbits
Chuanxin YU ; Xiuxiu GAO ; Zicheng WAN ; Wang LIANG ; Xiao WANG
Chinese Journal of Comparative Medicine 2017;27(7):53-58
Objective To observe the effects of road transport on hematological and biochemical parameters in New Zealand rabbits.Methods A total of 12 healthy New Zealand rabbits were selected for 2 h road transport.Blood samples were collected at 0, 24, 48, 72 and 96 h after transport, respectively.White blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), hematocrit (MCV), mean erythrocyte hemoglobin content (MCH), mean erythrocyte hemoglobin concentration (MCHC) and platelets (PLT) were measured using a blood analyzer.Blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), total protein (TP), urea nitrogen (UREA), creatinine, uric acid (UA), triglycerides (TG), total cholesterol (COHL), glucose (GLU), hypersensitive C-reactive protein (CRP), α-amylase (AMYL), and creatine kinase (CK) were detected by an automatic biochemical analyzer.Results Compared the parameters before and after transport, The WBC count was increased first (P< 0.05 or P < 0.01) and then decreased after transport, the levels of RBC, HGB, HCT and PLT were decreased first (P< 0.05 or P < 0.01) and then increased after transport, and MCV was significantly high at 96 h after transport (P< 0.05).Among the clinical biochemical parameters, ALT, AST and BUN were firstly elevated (P< 0.05 or P < 0.01) and then decreased.TP, ALB as well as CREA and TG were firstly decreased (P< 0.05 or P < 0.01) and then increased.GLU was significantly low at 24 h after transport (P< 0.05).All parameters except MCV at 96 h after transport were not significantly different from those before transport.Conclusions Changes of blood routine, liver and kidney function indexes, lipid metabolism indexes, glucose metabolism index and creatine kinase index are observed in the New Zealand rabbits after 2-hour road transportation, and all the indicators except MCV return to pre-transport levels within 96 h.
4.Rapid Determination of Long-chain Aliphatic Aldehyde in Gutter Oil by Fluorescent Derivatization-High Performance Liquid Chromatography
Wan WANG ; Qingfeng YU ; Ying XIAO ; Hua HE
Chinese Journal of Analytical Chemistry 2017;45(5):770-776
Reuse of waste oil in cooking has been the social concern for long time, but until now there is no reliable method to identify the gutter oil.In this work, based on the generation of long-chain aliphatic aldehydes in the refinement of gutter oil, gutter oil and adulterate cooking oil were identified by determining hexanal, heptanal, octanal, nonanal and decanal in oil sample with high perfomance liquid chromatography (HPLC) method using O-(3-(9H-carbazol-9-yl)propyl)hydroxylamine as the fluorescent derivative reagent.At first, 10 μL of oil sample was dissolved in 200 μL of isopropanol, then reacted with O-(3-(9H-carbazol-9-yl)propyl)hydroxylamine to form the stable derivatives, extracted with acetonitrile and injected into HPLC for analysis.The derivatives were separated on a C18 column within 15 min with ACN-H2O (90∶10, V/V) as mobile phase.The fluorescence detector was set at λex/λem=292 nm/348 nm.The linear range of 5 kinds of long-chain aliphatic aldehydes was 0.01-1.00 μmol/L with limit of detection (LOD, S/N=3) of 0.05-0.10 nmol/L, and the spiked recoveries were 95.6%-101.4%.The results showed that the edible oils obtained from supermarket almost had no hexanal, heptanal, octanal, nonanal, decanal, but these aldehydes increased significantly in gutter oil.Thus the 5 kinds of long-chain aldehydes could be used as the indictor of gutter oil.Taking the advantage of good specificity, higher sensitivity, good accuracy and simplicity, the method was suitable for the rapid identification of gutter oil.
5.Research on Pretreatment Method of Styrene Type Macroporous Resin and Its Application on Separation and Purification of Traditional Chinese Medicine
Jianxiong WU ; Qin WAN ; Jianping QING ; Zhenzhong WANG ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(8):1831-1836
This article was aimed to study pretreatment method of styrene type macroporous resin and its application in the industrial production. The organic residues and heavy metals of styrene type macroporous resin were detected respectively by gas chromatography and atomic absorption spectrophotometry before and after pretreatment to optimize the pretreatment process. The results showed that contents of organic residues and heavy metals of styrene type macroporous resin with the preferred method of pretreatment are in line with relevant regulations. It was concluded that this pretreatment method was simple and feasible, which is suitable to be applied in the industrial production.
6.Estrogendecreasingmatrixmetalloproteinase-9geneexpressioninmicewithexperimental autoimmune encephalomyelitis
Xiao HU ; Jiaying ZHU ; Weikun HUANG ; Xing WAN ; Jianyi WANG
Chinese Journal of Neurology 2011;44(7):468-472
Objective To study the regulation effect of estrogen in expression of matrix metalloproteinase-9 (MMP-9) in the central nervous system (CNS) in mice with experimental autoimmune encephalomyelitis ( EAE).Methods The 60 mice were overiectomized and 2 weeks later EAE was induced with MOG35-55 peptide in these mice.They were divided into a treatment group and a control group.The treatment group was treated with estrogen and the control group was given PBS.Clinical symptoms in these two groups were scored and compared.HE staining was used to observe inflammation in the brain and spinal cord.The MMP-9 expression in the CNS was examined by quantitative real-time PCR and immunofluorescence staining.Results The incidence of disease was lower (treatment and control group were 8/30 and 28/30 respectively) and clinical symptoms were milder (treatment and control group were 3.23±0.83 and 1.62 ±1.00 respectively,t=3.811 and P<0.05) in the treatment group than those in the control group.HE staining showed the decreased infiltration of inflammatory cell in the treatment group (Treatment group:inflammatory score were 0.895 ±0.206,0.752 ±0.302,0.732 ±0.183 in acute,relief and chronic phase respectively;Control group:inflammatory score were 3.472 ±0.635,2.881 ±0.662,1.891 ± 0.482 in acute,relief and chronic phase respectively.t = 8.622,6.543 and 5.027,all P < 0.05).The quantitative real-time PCR and immunofluorescence staining showed that the expression of MMP9 in the CNS was decreased in the treatment group.Conclusion Estrogen may decrease MMP-9 expression in the CNS,reduce inflammation and clinical symptoms in mice with EAE.
7.Exploration of the effect of Helq gene on stem cell pluripotency
Cong WAN ; Yaping HUANG ; Mei WANG ; Yamei XIAO ; Xiaoyang ZHAO
Acta Laboratorium Animalis Scientia Sinica 2016;24(4):358-363
Objective To explore whether Helq deletion affect the pluripotency of stem cells. Methods Helq knockout embryonic stem cells were obtained by CRISPR?Cas9 gene editing technique. Results The results of immunoflu?orescence analysis showed that the expression of Oct4 and Nanog had no obvious difference to that of the control cells. The Helq-/ - embryonic stem cells could produce viable pups by tetraploid complementation, indicating that their pluripotency was not affected. Meanwhile, we found that day 2 epiblast?like cells also were obtained through differentiation of the Helq-/ - embryonic stem cells in vitro. Immunostaining and real?time PCR analysis showed that the gene expression of Helq-/ - epiblast cells were similar to the wild type cells. Conclusions Taken together, it is proved that the genomic in?stability caused by Helq deletion does not affect the pluripotency of pluripotent stem cells.
8.Association between Expression of HDAC1 Protein and HPV16/18 Infection in Cervical Intraepithelial Neoplasia and Cervical Squamous Cell Carcinoma
Li XIAO ; Yizeng WAN ; Bao WANG ; Li LI
Journal of China Medical University 2015;(5):464-467,471
Objective To explore the anomalous expression of HDAC1 in cervical intraepithelial neoplasia(CIN)and cervical carcinoma and its relationship with the infection of HPV16/18. Methods The expressions of HDAC1 protein and HDAC1 infection were detected by in situ hybridiza?tion(ISH)and immunohistochemistry staining(SP)in tissues from patient with chronic cervicitis(the chronic cervicitis group),CIN(the CINⅠgroup,the CINⅡgroup and the CINⅢgroup)and cervical squamous carcinoma(the cervical carcinoma group). Results The infection rate of HPV16/18 was 8.33%,34.78%,48.00%,65.21%,78.00%in patients with chronic cervicitis,CINⅠ,CINⅡ,CINⅢand cervical carcinoma re?spectively. There was significant difference between groups(P<0.05). The positive expression rate of HDAC1 was 0,13.04%,32.00%,47.82%and 76.00%in chronic cervicitis,CINⅠ,CINⅡ,CINⅢand cervical carcinoma,respectively. The differences were statistically significant(P<0.05). The positive expression of HDAC1 in the HPV16/18 infected patients was significantly higher than those uninfected with HPV16/18(P<0.05). The HDAC1 positive expression had no relationship with clinical stages of cervical carcinoma,however,it was related to the tissue differentia?tion,stromal invasion and pelvic lymph node metastasis(all P<0.05). Conclusion The infection of HPV16/18 may play an important role in the carcinongenesis of SCC through increasing the expression of HDAC1.
9.Identification of the metabolites of Dingzhi Xiaowan extract in depressive rat plasma, urine, feces and bile after intragastric administration.
Lu XU ; Wan-wan LIU ; Xiao TAN ; Shi WANG ; Li-hua MU ; Xian-zhe DONG ; Dong-xiao WANG ; Ping LIU
China Journal of Chinese Materia Medica 2015;40(11):2214-2220
Dingzhi Xiaowan is a widely used traditional Chinese medicine in treating depression, which is a similar formula of Kaixinsan. In this research, a rapid ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS(E)) method was established to analyze the metabolites of Dingzhi Xiaowan in depressive model rat plasma, bile, urine and feces. After we established Chronic unpredictable mild stress (CUMS) model rats and orally administrated Dingzhi Xiaowan, rat plasma, bile, urine and feces samples were collected and prepared. Using Waters Cortects UPLC C18 column (2.1 mm x 50 mm, 1.6 μm), acetonitrile-0.1% formic acid mobile phase gradient, these samples were analyzed and 33 metabolites of nine bioactive compounds were detected and tentatively identified by Metabolynx. Among the 33 metabolites, three metabolites were identified from plasma sample, three came from bile sample, and 27 metabolites were identified from urine and feces samples. This approach provided a rapid method for characterizing the metabolites of Dingzhi Xiaowan and gave the truly active structures and the action mechanism of their antidepressant effects.
Animals
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Bile
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metabolism
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Chromatography, High Pressure Liquid
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Depression
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metabolism
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Disease Models, Animal
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Drugs, Chinese Herbal
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metabolism
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Feces
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chemistry
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Male
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Mass Spectrometry
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Medicine, Chinese Traditional
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Plant Extracts
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metabolism
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Rats
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Rats, Sprague-Dawley
10.Study of screening nephroprotective bioactive substances based on triple-color fluorescence probes in Carthami flos.
Xiao-Hong LAN ; Shun XIAO ; Wan GONG ; Yi WANG ; Xiao-Ping ZHAO
China Journal of Chinese Materia Medica 2014;39(10):1880-1885
In this study, an approach based on triple-color fluorescence probes was developed for screening potential nephro-protective bioactive substances. Three fluorescent probes (i. e. FDA, MTR and Hoechst 33342) were used to label HK-2 cells injured by doxorubicin hydrochloride, and cellular fluorescence images were subsequently acquired and analyzed by a cellular-fluorescence image microscopy platform. The established method was applied to screening 53 components of Carthami Flos, and three components C17, C18 and C19 were found to exhibit nephroprotective effects against doxorubicin hydrochloride induced injury on HK-2 cells. Eight compounds (i. e. hydroxysafflor yellow A, 6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside, 6-hydroxykaempferol-3,6-di-O-gluco-side or 6-hydroxykaempferol-6, 7-di-O-glucoside, 6-hydroxykaempferol-3-O-rutinoside, 6-hydroxykaempferol-3-O-glucoside or 6-hydroxykaempferol-7-O-glucoside, rutin, isoquercetin, and kaempferol-3-O-rutinoside) in components C17, C18 and C19 were preliminarily identified by liquid chromatography-mass spectrometry (LC-MS). Isoquercetin, rutin, kaempferol-3-O-rutinoside, and hydroxysafflor yellow A were confirmed by comparing with reference substances, Further study indicated that these four compounds had moderate nephroprotective effects, while isoquercetin showed a significant nephroprotective effect in a dose-dependent manner. These results suggest that isoquercetin, rutin, kaempferol-3-O-rutinoside and hydroxysafflor yellow A might be the nephroprotective bioactive substances in Carthami Flos.
Carthamus
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chemistry
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Cell Line
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Chromatography, High Pressure Liquid
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Dose-Response Relationship, Drug
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Drug Evaluation, Preclinical
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Flowers
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chemistry
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Fluorescent Dyes
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chemistry
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Humans
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Kidney
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chemistry
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cytology
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drug effects
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Protective Agents
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chemistry
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pharmacology