2.Extraction and isolation of polysaccharide from ginseng and its anti-tumor activity in vitro
Ming REN ; Xiaoshi HAO ; Lingyan YE ; Wanze ZHANG ; Yanfei QI ; Kun XU ; Juan LI
Journal of Jilin University(Medicine Edition) 2014;(4):812-815
Objective To extract,isolate and identify the polysaccharide from ginseng,and to investigate its anti-tumor activity in vitro. Methods The ginseng polysaccharide was obtained through water extraction and ethyl alcohol deposition method. Use the Sevage method to remove the protein in the crude polysaccharide. The structural characteristics of the polysaccharide were determined by FT-IR spectra.The RM-1 and HeLa cells were divided into control group and different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )of ginseng polysaccharide groups. The survival rates of the cells in various groups were detected by MTT method. The indirect killing effects of ginseng polysaccharide with different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )on the cancer cells were determined by CTL test. Results The extraction rate of ginseng polysaccharide was 8.7 6% and the structural characteristics demonstrated that the main component of the extract was polysaccharide.The result of MTT showed that there were no significant differences of the survival rates of RM-1 and HeLa cells between different concentrations of ginseng polysaccharide groups after treated for 24 h compared with control group (P>0.05).The result of CTL test showed that the cytototic LHD release rates in different concentrations of ginseng polysaccharide groups were increased significantly (P<0.05)compared with control group;with the increase of ginseng polysaccharide concentration, the cytotoxic LDH release rates were increased firstly and then were decreased.When the concentration of ginseng polysaccharide was 100 mg·L-1 ,the cytotoxic LDH release rate was the biggest.Conclusion Ginseng polysaccharide can indirectly inhibit the growth of the tumor cells by activating T cells and play an anti-tumor effect.
3.Preparation of polysaccharide complex and its indirect antitumor activity in vitro
Lingyan YE ; Ming REN ; Lin LYU ; Li LI ; Yanfei QI ; Juan LI ; Kun XU
Journal of Jilin University(Medicine Edition) 2014;(5):1033-1037
Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.
4.Ginkgolide B inhibits apoptosis in high glucose-stimulated human umbilical vein endothelial cells
Kun CHEN ; Ming ZHANG ; Beidong CHEN ; Yanyang ZHAO ; Wei WU ; Ruomei QI
Chinese Pharmacological Bulletin 2017;33(3):378-383
Aim Toinvestigatetheeffectofginkgolide B on apoptosis in high glucose-treated endothelial cells.Methods Humanumbilicalveinendothelial cells(HUVECs)were used in the present study.The level of transmigration of HUVECs was analyzed by Tr-answell experiment.Apoptosis was detected by flow cy-tometry.Reactive Oxygen Species (ROS ) was meas-ured by immunofluorescence kit.The protein expres-sionwasanalyzedbyWesternblot.Result Highglu-cose treatment resulted in a reduction in transmigration of HUVECs and ginkgolide B recovered the phenome-non in glucose-treated endothelial cells.The level of ROS generation was increased in high glucose-treated group,whereas ginkgolide B inhibited ROS genera-tion.Immunofluorescence data showed high glucose in-creased apoptosis,whereas ginkgolide B inhibited ap-optosis in high glucose-treated HUVECs.Moreover, the expressions of Bax and caspase-3 were increased and Bcl-2 was reduced in high glucose-treated group. In contrast,ginkgolide B abolished the expressions of Bax and caspase-3 and increased Bcl-2 expression. Moreover,high glucose enhanced the expression and phosphorylation of p53,while ginkgolide B suppressed the expression and phosphorylation of p53 induced by highglucose.Conclusions GinkgolideBcaninhibit apoptosis and improve transmigration function in high glucose-treated HUVECs.Ginkgolide B has protection against high glucose-induced endothelial cell injury.
5.Effects of breviscapine on epithelial-mesenchymal transition induced by transforming growth factor-β2 in human lens epithelial cells
Kun-ming, CUI ; Feng-yan, ZHANG ; Ying, QI ; Meng-meng, WANG ; Hang, GAO
Chinese Journal of Experimental Ophthalmology 2013;31(10):930-934
Background Epithelial-mesenchymal transition (EMT) is a major event in the pathogenesis of posterior capsular opacification (PCO),and the expressions of α-smooth muscle actin (α-SMA) is the marker of EMT.Previous studies showed that breviscapine plays an important role in anti-fibrosis and suppression EMT,however,the mechanism of its effect on EMT in LECs is unclear.Objective Present study was to investigate the effect of breviscapine on the expression of α-SMA and fibronectin (FN) in human LECs induced by transforming growth factor-β2 (TGF-β2).Methods Human LECs strain,HLE-B3,was cultured and passaged in DMEM containing 10% fetal bovine serum.Different concentrations of breviscapine (6.75,12.75,25.00,50.00 and 100.00 mg/L)were added into the medium for 24,48 and 72 hours respectively,and then cell cunting kit-8 (CCK-8) was used to evaluate the half maximal inhibitory concentration (IC50) of breviscapine to HLE-B3.In addition,HLE-B3 was subcultured at the density of 1 × 106/hole and divided into 4 groups.The cells were in free-serum medium as the normal control;the cells were exposed in 10 μg/L TGF-β2 as TGF-β2 group;while in the breviscapine group,10 mg/L of breviscapine was added into the culture medium and another group was the combination of 10 mg/L breviscapine and 10 μg/L of TGF-β2 treatment.Real-time PCR and Western blot were used to detect the mRNA expressions of α-SMA and FN as well as their protein in HLE-B3 72 hours after cultured.Results The inhibitory rate of breviscapine to HLE-B3 proliferation was gradually elevated with the increase of concentration of breviscapine,showing a significant inhibition of the cell proliferation among the different groups and at various time points (F =292.851,P=0.000;F =65.037,P=0.000).IC50 of HLE-B3 at 72 hours was 22 mg/L,and therefore,in rest of experiment 10 mg/L of breviscapine was used which was 1/2 only half of the IC50.α-SMA and FN were expressed in cultured normal HLE-B3.The expressing level of α-SMA mRNA and FN mRNA in the HLE-B3 was significantly different among the normal group,TGF-β22 group,breviscapine treatment group and combination group as well as at various time points (F =105.490,P =0.000 ; F =1041.414,P =0.000).Similarly,the protein expressions of α-SM A and FN in the HLE-B3 was significantly different among the four groups and different time points (F=136.872,P=0.000;F=119.820,P=0.000).The expression levels of α-SMA and FN mRNA and their proteins in HLE-B3 were remarkably increased in the 10 μg/L TGF-β2 group compared with the normal control group (at all P=0.000),and those in the combination group were obviously declined in comparison to the TGF-β2 group (P =0.001,0.001,0.001,0.010).No significant difference was found in the expressions of α-SMA and FN in the HLE-B3 between the breviscapine group and normal control group in both transcriptional level and protein level (P =0.551,0.292,0.551,0.360).Conclusions 10 mg/L breviscapine can arrest the proliferation and EMT of human LECs.This result suggests that using breviscapine may be a potential prophylactic approach in the prevention of PCO.
6.The effect of magnitopuncture stimulation on HRV during simulated driving under vibration conditions.
Zengyong LI ; Kun JIAO ; Ming CHEN ; Chengtao WANG ; Shaohua QI
Journal of Biomedical Engineering 2003;20(1):97-100
The purpose of this study was to assess the effect of magnitopuncture stimuli for reducing driver mental stress and fatigue using power spectral analysis of the heart rate variability (HRV) and subjective evaluation. The experiments were divided into A-group and B-group. In both groups the subjects performed the simulator for 90 minutes under a vibration conditions with an erect sitting posture in a silent environment, and magnitopuncture was put on the acupoints when performing the task for one hour in A-group. In this study HRV exhibited a significant difference between the two groups after the simulating task (P < 0.05). A conclusion that magnitopuncture stimuli can reduce the driver mental stress and fatigue effectively was drawn.
Acupuncture Points
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Adult
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Automobile Driving
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Autonomic Nervous System
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physiopathology
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Electric Stimulation Therapy
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methods
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Fatigue
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therapy
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Heart Rate
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physiology
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Humans
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Magnetics
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therapeutic use
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Male
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Stress, Physiological
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therapy
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Vibration
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adverse effects
7.Therapeutic effect of ulinastatin on traumatic brain edema accompanied by seawater drowning in rats
Xiang-Qian QI ; Ming-Kun YU ; Yan DONG
Chinese Journal of Neuromedicine 2008;7(11):1142-1146
Objective To observe the therapeutic effect of ulinastatin on traumatic brain edema (TBE) accompanied by pulmonary edema due to seawater drowning in rats. Methods Thirty-two Sprague-Dawley rats were randomly divided into control group (nffi8) and ulinastatin treatment group (n=24). A rat model of moderate brain trauma was established by lateral head impact, and pulmonary edema was induced in these rats by pulmonary lavage with seawater to mimic seawater drowning. Twenty-four hours after intmperitoneal injection of ulinastatin, the changes in the cerebral and pulmonary water contents and concentrations of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in the brain, lungs and serum were measured, and the histopathological changes of the brain and lung tissues were observed. Results The cerebral and pulmonary water contents and the IL-1β and TNF-α concentrations in the serum, brain and lungs of the rats with brain trauma and pulmonary edema were markedly decreased after ulinastatin injection, which also resulted in obvious improvement of the brain and lung pathologies induced by the injuries. Conclusion Ulinastatin can alleviate traumatic brain edema in rats with pulmonary edema due to seawater drowning by inhibiting the proinflammatory cytokines.
8.Relationship between selection of Pinus massoniana families and Folium Pini.
Man-xi ZHAO ; Cui-qi YAN ; Wei WANG ; Jian-ming YE ; Yong-kun ZHONG ; Zun-hong KE ; Xiao-feng HAO ; Xiao KE ; Liang YE ; Lu-qi HUANG
China Journal of Chinese Materia Medica 2015;40(9):1699-1704
Based on variation of Pinus massoniana families, heritablility and correlation analysis, the contents of shikimic acid and procyanidine (heritability 0.90, 0.70), dry weight of single branch (heritability 0.60) and and leaf length (heritability 0.46) were screened out as quality, yield and harvest cost traits of Folium Pini, respectively. For the different medicinal application of Folium Pini, varied methods were chosen to estimate weight and construct index equation. Weight adjustment based.on equal emphasis were used as economic weight determining method to select the best families, and the index (accuracy 0. 936 4 and heritability 0. 881 6) obtained was a little better than that obtained by equal emphasis, and much better than that by restricted index. The superior families selected with adjustment weight and equal emphasis were No. 46, 43 and 28. Partial regression were used as economic weight determining method to select the best families,and the index obtained had the highest accuracy (0.941 5) , index heritability (0. 889 9) and the genetic gain of shikimic acid content. The superior families selected with this method were No. 46, 27 and 47. No. 46 was the best families with maximal economic benefit. Our study indicated that suitable method for estimate weight and construct index equation can be applied for better accuracy of superior families selection of P. massoniana.
Breeding
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Drugs, Chinese Herbal
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analysis
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Pinus
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chemistry
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classification
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genetics
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growth & development
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Plant Leaves
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chemistry
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Plants, Medicinal
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chemistry
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classification
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genetics
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growth & development
9.High Expression, Purification and Immuno-indentification of Recombinant HIV Surface Antigen gp120
Chen, CHEN ; Kun, LIU ; Jin-zhong, WANG ; Ming, MA ; Yun-qi, CENG ; Qi-min, CHEN ; Xiao-ke, ZHU ; Jian-hui, XIA
Virologica Sinica 2001;16(2):109-113
Different-length fragments of the HIV trans-membrane antigen (gp120) gene were expressed in Escherichia coli, using T7 expression system. SDS-PAGE stained by Coomassic Brilliant Blue showed no expression of full-length gp120 and poor expression of half-length gp120 fragments from the N-terminal, but high expression of 1/3-lgenth gp120 gene fragments from the N-terminal (including V1/V2 epitopes), more than 18% of total bacterial protein. Western blot showed fairly good reactivity to serum from HIV-infected individual. On this basis, we expressed the corresponding tp 120 fragments of the HIV strains epidemic in China. This study laid a solid foundation for exploration of high expression of gp 120 in E.coli and development of HIV serological diagnostic system for Chinese people.
10.The effects of compound CX09040 on the inhibition of PTP1B and protection of pancreatic β cells.
Ran-qi TANG ; Xiao-lin ZHANG ; Jin-ying TIAN ; Si-ming KONG ; Ying ZHOU ; Pei ZHANG ; Hong-kun YANG ; Song WU ; Ying ZHANG ; Fei YE
Acta Pharmaceutica Sinica 2015;50(6):682-689
To investigate the effects of 2-(4-methoxycarbonyl-2-tetradecyloxyphenyl)carbamoylbenzoic acid (CX09040) on protecting pancreatic β cells, the β cell dysfunction model mice were induced by injection of alloxan into the caudal vein of ICR mice, and were treated with compound CX09040. Liraglutide was used as the positive control drug. The amount and the size of islets observed in pathological sections were calculated to evaluate the β cell mass; the glucose stimulated insulin secretion (GSIS) test was applied to estimate the β cell secretary function; the oral glucose tolerance test (OGTT) was taken to observe the glucose metabolism in mice; the expressions of protein in pancreas were detected by Western blotting. The effects on the target protein tyrosine phosphatase 1B (PTP1B) were assessed by the PTP1B activities of both recombinant protein and the intracellular enzyme, and by the PTP1B expression in the pancreas of mice, separately. As the results, with the treatment of CX09040 in alloxan-induced β cell dysfunction mice, the islet amount (P<0.05) and size (P<0.05) increased significantly, the changes of serum insulin in GSIS (P<0.01) and the values of acute insulin response (AIR, P<0.01) were enhanced, compared to those in model group; the impaired glucose tolerance was also ameliorated by CX09040 with the decrease of the values of area under curve (AUC, P<0.01). The activation of the signaling pathways related to β cell proliferation was enhanced by increasing the levels of p-Akt/Akt (P<0.01), p-FoxO1/FoxOl (P<0.001) and PDX-1 (P<0.01). The effects of CX09040 on PTP1B were observed by inhibiting the recombinant hPTP1B activity with IC50 value of 2.78x 10(-7) mol.L-1, reducing the intracellular PTP1B activity of 72.8% (P<0.001), suppressing the PTP1B expression (P<0.001) and up-regulating p-IRβ/IRβ (P<0.01) in pancreas of the β cell dysfunction mice, separately. In conclusion, compound CX09040 showed significant protection effects against the dysfunction of β cell of mice by enlarging the pancreatic β cell mass and increasing the glucose-induced insulin secretion; its major mechanism may be the inhibition on target PTP1B and the succedent up-regulation of β cell proliferation.
Alloxan
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Animals
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Benzoates
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pharmacology
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Biological Assay
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Disease Models, Animal
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Glucose
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metabolism
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Glucose Tolerance Test
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Insulin
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secretion
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Insulin Resistance
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Insulin-Secreting Cells
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drug effects
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Liraglutide
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pharmacology
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Mice
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Mice, Inbred ICR
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Molecular Weight
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Pancreas
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drug effects
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enzymology
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Protein Tyrosine Phosphatase, Non-Receptor Type 1
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antagonists & inhibitors
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Signal Transduction