3.Extraction of diosgenin from Dioscorea zingiberensis
Feng HAN ; Wenhong LI ; Dong LI ; Xuan TANG ; Lianhong CHEN
Chinese Traditional Patent Medicine 1992;0(12):-
AIM:To study the application of ultrasonic extraction in process of extracting diosgenin from Dioscorea zingiberensis C.H.Wright,and to evaluate its merits.METHODS:A new process of ethanol extracting and hydrochloric acid hydrolysis was provided to extraction diosgenin,and five different extraction methods were studied and compared,and the reaction mechanism was discussed.RESULTS:The ultrasonic extraction method was more efficient.Compared with the traditional water-hydrolylis,extraction rate was 15 percent higher,dosage of hydrochloric acid was registered 90 percent decrease,and the production was purer.CONCLUSION:The grinding pretreatment of Dioscorea zingiberensis is help to enhance the effects of ultrasonic extraction,the method has some advantages in efficiency,energy-saving and low pollution.
4.Protective effects of thymosin β4 on ethanol-induced rabbit corneal epithelial cell damage in vitro
Zhixin, JIANG ; Peng, HAO ; Xin, TANG ; Xuan, LI
Chinese Journal of Experimental Ophthalmology 2016;34(2):108-114
Background Laser assisted subepithelial keratomileusis (LASEK) is one of surgical procedures for refractive correction.Dilute alcohol that is used for the removal of epithelium during LASEK induces the apoptosis of corneal epithelial cells.Researches showed that thymosin β4 (Tβ4) can arrest apoptosis, but whether Tβ4 plays inhibitory effect on ethanol-induced damage of corneal epithelial cells is still unelucidated.Objective The aim of this study was to investigate the protective effects of Tβ4 on ethanol-induced rabbit corneal epithelial cell damage in vitro.Methods The corneal tissue of deendothelium was obtained from 10 New Zealand white rabbits.Corneal epithelial cells were cultured in vitro by using explant culture method.Cultured cells were identified by detecting the expression of keratin 12 and connexin 43 with reverse transcription PCR (RT-PCR).The cells of second generation were collected and divided into 4 groups.The cells were regularly cultured in the normal control group, and Tβ4 was added in the culture medium at the final concentration of 1 μg/ml in the Tβ4 treated group.Ethanol-induced rabbit corneal epithelial cell damage models were established by adding PBS containing 20% alcohol in medium for 20 seconds in the model group,and Tβ4 was added in the medium of model cells at the final concentration of 1 μg/ml in the model+Tβ4 group.The survival rate of the cells was detected by MTT assay, and the apoptosis rate of the cells was examined by TUNEL method.The relative expression levels of cyclin D1 mRNA and cyclin-depensent protein kanase 4 (CDK4) mRNA in the cells were detected by real-time flurescenee quantitative PCR.The content of bcl-2 protein in the cells was detected by ELISA assay.Spectrophotometry was employed to measure the activity of caspase-3.The study complied with the Regulation for the Adminstration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The mean cell survival rate was (52.1 ± 14.07) % in the model group,which was significantly reduced in comparison with 100% of the normal control group and (77.7± 19.60) % of the model+Tβ4 group (P=0.001 ,P=0.035).TUNEL staining revealed more positive cells in the model group and the model+Tβ4 group,and the percentage of TUNEL positive cells was (30.0±6.7)% in the model+Tβ4 group, showing an evident decline in comparison with (42.4±4.0)% of the model group (P=0.01).The relative expression levels of cyclin D1 mRNA and CDK4 mRNA were 0.93±0.17 and 0.88±0.09 in the model+Tβ4 group, which were significantly higher than 0.68±0.05 and 0.54±0.07 in the model group (P=0.027,0.002).ELISA assay exhibited that bcl-2 content in the cells was considerably lower,and caspase-3 activity was significantly higher in the model group than those in the model+Tβ4 group (P =0.030,0.021).Conclusions Tβ4 plays a protective effect on rabbit corneal epithelial cells from apoptosis by lowing the caspase 3 activity and increasing bcl-2 content in ethanoldamaged rabbit corneal epithelial cells.In addition, Tβ4 promotes the regrowth of corneal epithelial cells by upregulating the expression of cyclin D1 and CDK4.
5.Research delta opioid receptor and its association with hepatocellular carcinoma
Xuan LI ; Bo TANG ; Jing ZHANG ; Xingsi LIANG ; Yangchao WEI
Chinese Journal of Hepatobiliary Surgery 2016;22(11):790-792
Opioid receptors have drawn highly attention to scientists since they were founded in the 1990s.As a member of G protein-coupled receptor family,opioid peptides are the endogenous ligands.It is well known to us that the basic structure consists of extracellular amino residues,seven transmembrane region and the hydroxyl residues of the cell.In recent years,the progress of science laid the foundation for further studies on the function of delta receptor.Aberrant expression of delta opioid receptor in a variety of tumors,and it plays an important role of the occurrence and development of tumors.This review describes the recent research advances on delta opioid receptor and its association with hepatocellular carcinoma.
6.The soluble expression and identification of single-chain fragment V antibodies against SSA antigen epitopes from the pHEN2 phagemid library
Hongbin LI ; Xuan ZHANG ; Fulin TANG ; Fengchun ZHANG
Chinese Journal of Internal Medicine 2010;49(7):614-617
Objective To obtain the soluble single-chain fragment V (ScFv)monoclonal antibodies (McAbs) against the SSA antigen epitopes.Methods Three octapeptides (60 000 SSA antigen residues 482-493 termed as P1 epitope, residues 310-323 termed as P2 epitope and residues 230-241 termed as P3 epitope) were synthesized on the lysine frame.The McAbs were panned by coating the corresponding as targets.The specificity, affinity and gene squences of the positive clones were assessed.Soluble single-chain fragment V antibodies special for SSA antigen epitopes were expressed and then identificated.Results After 5 rounds of panning, reactive scFv clones contained full-length scFv antibodies coding regions were obtained,with sufficient affinity and specificity for respective antigen peptides.The absorbance values at 410 nm of the fusion protein of anti-P1-P3 activity with the corresponding peptides were 1.43 ± 0.23, 0.82 ±0.31 and 0.80 ± 0.25, and there was also statistically significant difference in the cross reactions ( P < 0.01 ).Three clones were successfully expressed and then purified by His-bind resin.The activity in vivo of soluble ScFv antibodies was identified to be positive by the indirect immune-fluorescence assay on Hep-2 cells.Conclusion Souble ScFv McAbs against corresponding SSA antigen peptides with high affinity,specificity and activity in vivo were obtained, which are to be competent enough for epitopes expression on the target organs.
7.De novo sequencing and analysis of root transcriptome to reveal regulation of gene expression by moderate drought stress in Glycyrrhiza uralensis.
Chun-rong ZHANG ; Xue-yu SANG ; Meng QU ; Xiao-min TANG ; Xuan-xuan CHENG ; Li-ming PAN ; Quan YANG
China Journal of Chinese Materia Medica 2015;40(24):4817-4823
Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts
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Gene Expression Regulation, Plant
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Glycyrrhiza uralensis
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genetics
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Plant Roots
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Sequence Analysis, DNA
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Stress, Physiological
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Transcriptome
8.Establishment of an intracerebrai hemorrhage model under the ultrasonic guidance
Xuan ZHOU ; Li CHEN ; Bei LI ; Cong FENG ; Jie TANG ; Aijun LIU ; Tanshi LI ; Faqin Lü
Chinese Journal of Ultrasonography 2012;21(4):340-343
Objective To explore a method of establishing a canine model of intracerebral hemorrhage (ICH) by puncturing the main branch of middle cerebral artery (MCA) under ultrasonic guidance and proceed a pilot study of contrast-enhanced ultrasound (CEUS) of ICH.Methods Twelve adult healthy mongrel dogs were enrolled in the experiment.In the condition of bone window,the main branch of MCA was punctured to bleed under the ultrasonic guidance.CEUS of brain was performed every 30 minutes until hematoma formed and another CEUS was performed at 24 hours.Then brains were collected for pathologic examination after dogs were euthanized.Results Eleven of twelve dogs developed ICH successfully that located in the Out edge of basal ganglia,which was confirmed by CEUS and autopsy.One dog died within 6 hours due to intraventricular hemorrhage which was confirmed by autopsy.The hematoma diameters were measured as (22.4 ± 7.1)mm by CEUS before euthanasia and as (21.6 ± 6.9)mm on autopsy.There was no significant difference between the two methods of measurement (t =1.521,P =0.1565).The characteristics of active bleeding and hematoma on ICH were performed in the CEUS imaging.Conclusions A model of dog ICH by puncturing the main branch of MCA under the guidance of ultrasound was established successfully,and it proved to be simple,effective and repeatable.The imaging characteristics of this model are in good accordance with those of ICH in patients.
9.Risk factor analysis of diabetic retinopathy based on community health management files
Jie, CHEN ; Zhong-Ning, XUAN ; Bin-Yi, LI ; Rong, LIN ; Yin, TANG
International Eye Science 2016;16(6):1180-1182
?AIM:To investigate and analyze diabetic retinopathy ( DR) in patients with type 2 diabetes in local community and risk factors of type 2 diabetic retinopathy, and to provide reference for the prevention and treatment of diabetic.?METHODS:Randomly 268 cases with type 2 diabetes from community health management files from January to March 2015 were selected. Fundus photography was read through the remote reading system, demographic data and laboratory indexes of DR were analyzed.?RESULTS:In the 268 cases with type 2 diabetes, 85 cases were diagnosed as DR ( 31. 7%); there were statistical difference on gender, duration, systolic blood pressure, fasting glucose, glycosylated hemoglobin A1c, urine creatinine between patients with DR and without ( P<0. 05 ). Multivariable Logistic regression analysis showed, duration, systolic blood pressure, fasting glucose, glycosylated hemoglobin A1c were independent risk factors of DR(P<0. 05).?CONCLUSION:Long duration, high blood pressure, high blood glucose are main risk factors of DR, we should strengthen the monitoring of blood pressure, blood glucose and prevent the happening of the DR.
10.Clinical Application of Ultrasonography on Tethered Spinal Cord Syndrome
ying-liang, LI ; yi, TANG ; zuo-zhong, XIA ; ping, LIANG ; xuan, ZHAI
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To explore the clinical application of ultrasonography on tethered spinal cord syndrome (TCS).Methods Ultrasonic feature of 45 patients with TCS were analyzed retrospectively. Pre-and post-operative blood flow rates of cone were recorded by color doppler.Results Ultrasound was the same as computer tomography not only presented the image of anatomy and pathology of TCS,but also showed the lack of pulsatile motion of distal cord with TCS.Before the operation,blood flow rates of cone were( 0.047?0.012) m/s.After the operation,blood flow rates of cone were(0.158?0.029) m/s.There was significantly different(P