1.Study of glomerular podocyte injury induced by aristolochic acid
Hong CHENG ; Yipu CHEN ; Hongrui DONG ; Yanyan WANG ; Hongliang RUI
Chinese Journal of Nephrology 2012;28(3):222-225
Objective To explore whether the glomerular podocytes can be damaged by aristolochic acid. Methods Thirty-two male SD rats were equally divided into the following 2 groups:model group in which the rats received the extract of Aristolochia manshuriensis Kom (AmK) by gavage; control group only received tap water by gavage.24 h urinary protein excretion was measured at the end of the 1st and 4th week,and SDS-PAGE gel electrophoresis was performed to detect the protein in urine.At the end of the 4th week,all the rats were sacrificed and the glomeruli were isolated by laser capture microdissection technique.The mRNA expression of nephrin,podocin,CDA2P,podocalyxin and podoplanin in isolated glomeruli was determined by RT-PCR,and the average width of glomerular foot process was measured by electron microscopy and image analysis. Results At the end of the 4th week,24 h urinary protein excretion in the model group was significantly higher than that in the control group (P<0.01) and the urinary albumin content in model group was also obviously increased.The average width of glomerular foot process in the model group was significantly larger than that in control group (P<0.01).The mRNA expressions of nephrin,podocin,CDA2P,podocalyxin and podoplanin in glomeruli were significantly down-regulated in the model group compared with the control group,which decreased by 34%,62%,56%,50%(P<0.01) and 27% (P<0.05),respectively. Conclusions Aristolochic acid can damage the glomerular podocytes,resulting in the down-regulation of nephrin,podocin,CD2AP,podoplanin and podocalyxin mRNA expression, the segmental widening of foot process, and increased urinary protein excretion.
2.Effect of aldosterone receptor antagonist on obesity-related glomerulonephropathy
Jing DONG ; Hong CHENG ; Min YANG ; Yanyan WANG ; Hongrui DONG ; Hongliang RUI ; Yipu CHEN
Chinese Journal of Nephrology 2014;(12):919-924
Objective To examine whether aldosterone contribute to obesity related glomerular disease. Methods C57BL/6J mice were randomly divided into three groups: a control group (low?fat?diet, n=10), a model group (high?fat?diet, n=10) and a intervention group (high?fat?diet, n=12). After 8 weeks intervention group were treated with a mineralocorticoid receptor antagonist, spirolactone (SPL).The physicochemical indexes and the renal pathology of the three groups were all detected. The mRNA and protein expressions of podocyte marker protein were determined by real?time PCR and Western blotting, respectively. Results Compared with the control group, body weight, kidney weight, Lee ’s index, fat index, blood cholesterol, blood triglyceride, creatinine clearance rate, urinary protein excretion, glomerular average diameter, glomerular foot process average width were significantly up ? regulated (P<0.05); The mRNA and protein expression of nephrin, podocin, podoplanin and podocalyxin were significantly down?regulated in model group (P<0.05). Meanwhile, these changes were attenuated by SPL. Conclusion Aldosterone can participate in the process of obesity? related renal injury, and these can be attenuated by mineralocorticoid receptor antagonist, spirolactone. This gives us preliminary clues to treat ORG.
3.Evaluation of urine analysis by flow cytometry and strip test in diagnosing urinary tract infection
Fie QI ; Jian PAN ; Jiang HAN ; Shi CHENG ; Quan DONG ; Tingju ZHANG ; Rui MA ; Guijian LIU
Chinese Journal of Laboratory Medicine 2009;32(6):630-634
Objective To evaluate the clinical application of automated urine formed elements analyzer and/or urine dipstick analyzer for examination of urinary formed elements in screening urinary tract infection (UTI). Methods 148 fresh midstream clear-catch urine samples from the UTI patients and 284 fresh midstream clear-catch urine samples from non-UTI subjects were selected. Bacteria culture was performed for bacterial colony counting and identification. Bacteria counts ( BACT), yeast-like fungus and WBC were performed by UF-looOi automated urine formed elements analyzer. Leukocyte esterase test (LEU) and nitrite test (NIT) were performed by URISYS 2400 urine dipstick analyzer. We evaluated data obtained from urine dipstick analyzer, UF-1000i and combination of UF-1000i with urine dipstick analyzer and the results was compared with those obtained from quantitative bacterial culture. Then we evaluated the sensibility, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy. Results Among the 148 patients with UTI, the positive rate of the quantitative bacterial culture was 73.6% (109/148), the positive rate of LEU and NIT detected by dipstick test 26. 4% (39/148).There was significantly statistical difference between bacterial culture and strip test(χ2 = 55.68 ,P < 0. 05 ). The positive rate of urine flow cytometry by UF-1000i with either positive of BACT and WBC was 91.2%(135/148), which was higher than the positive rate of the quantitative bacterial culture. There was significant difference between two methods (χ2 = 14. 70, P < 0. 05 ). The positive rate of anyone positive among BACT, WBC, LEU and NIT was 94. 6% (140/148) when detected with combination of dipstick test and UF-1000i, which was higher than the positive rate of the quantitative bacterial culture. And there was significant difference between two methods (χ2 = 20. 45, P < 0. 05 ). The sensitivity of dipstick test was low (26. 4% ,39/148 ), and specificity was high ( 99. 3%, 282/284 ) . The sensitivity, specificity, positive predictive value, negative predictive value of BACT detected by UF-1000i in diagnosing urinary tract infection were 92. 6% ( 137/148 ), 39. 8% ( 113/284 ). 44. 5% ( 137/308 ) and 91.1% ( 113/124 ), respectively. If the dipstick test was combined with UF-1000i, the sensitivity, negative predictive value, specificity, positive predictive value and accuracy were 98.0% ( 145/148 ), 97.1% ( 100/103 ). 35.2% (100/284) ,44. 1% (145/329) and 56. 7% (245/432), respectively. Conclusions The combination of urine dipstick test and automated urine formed elements analyzer UF-1000i plays an important role in early diagnosis of UTI. And it has significant value in diagnosis of UTI, especially for the patients with negative bacterial cultures of urine sample.
4.Changes of Lead,Zinc,Copper,Iron and Calcium in Blood of Lead Poisoned Infantal Mice
rui-fang, HE ; yan, ZHANG ; yan-xu, YANG ; xiao-juan, LI ; yuan, CHENG ; dong-liang, LI
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To investigate the changes of lead,zinc,copper,iron and calcium in blood of chronic poisoned infantal mice.(Methods) Forty-eight 21 day-old kunzea mice were randomly divided into 4 groups,each having 12 mice.Distilled water group was as control group and other three lead acetate poisoning groups had a dose of 10,20,40 mg/kg,respectively.The poisoning was carried out by lavage once a day,and consecutively for 46 days.Eyeballs of mice were picked then for blood sampling,and BS trace element analysis grapher was used to determine level of lead,zinc,copper and iron.Level of calcium was measured by Dimentional-RXL auto-biochemistry analysis meter.Results The lead and zinc levels in poisoned mice blood were increased with increasing lead acetate level administration,while zinc level changed inversely with lead acetate level.Significant differences were shown among control group and poisoning groups in terms of lead(P0.05).Conclusion Lead posioning can lead to zinc decreasing and copper(increa)-sing,which suggests that zinc works as a poential antidote of lead poisoning.
5.Chemical constituents of leaves of Panax japonicus var. major.
Rui HE ; Qi LIU ; Yin-Huan LIU ; Jiang CHAI ; Dong-Dong ZHAO ; Wei WANG ; Jiu-Cheng CUI ; Xiao-Mei SONG ; Zheng-Gang YUE
China Journal of Chinese Materia Medica 2014;39(9):1635-1638
Seven compounds were isolated from the leaves of Panax japonicus var. major by chromatographic methods including silica gel, Sephadex LH-20, ODS and semi-preparative HPLC. Their structures were elucidated by their physical and chemical properties and spectral data analysis as 5, 7-dihydroxy-8-methoxyl flavone (1), ginsenoside Rs2 (2), quinquenoside R1 (3), ginsenoside Rs1 (4), notoginsenoside Fe (5), ginsenoside Rd2 (6) and gypenosiden IX (7). Among them, compound 1 was obtained from the Panax genus for the first time, and compounds 2-7 were isolated from this plant for the first time.
Chromatography, High Pressure Liquid
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Flavones
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analysis
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chemistry
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isolation & purification
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Ginsenosides
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analysis
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chemistry
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isolation & purification
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Magnetic Resonance Spectroscopy
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Panax
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chemistry
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Plant Leaves
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chemistry
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Spectrometry, Mass, Electrospray Ionization
6.Light and temperature and their effects on photosynthesis characteristics of stereoscopic cultivation in Panax notoginseng.
Yao-long WANG ; Xiu-ming CUI ; Lei LAN ; Wei-dong CHEN ; Rui-bo LI ; Cheng-xiao WANG ; Xiao-yan YANG ; Da-hui LIU ; Ye YANG
China Journal of Chinese Materia Medica 2015;40(15):2921-2929
Light intensity, gas temperature, soil temperature and gas exchange parameters were determined of three years old Panax notoginseng planted on different layers seedbed and different location (left, middle, right) of the same layer in greenhouse. Result show that diurnal variation of light intensity, gas temperature and soil temperature showed that upper layer > middle layer > lower layer; different locations of the same layer showed that light intensity of upper layer was not different among different locations; light intensity of middle and lower layer in right and left were the same, and significantly higher than those in the middle position; the gas temperature of each layer all with less different of each location; soil temperature of 12 cm depth is the lowest, and was gradually increased to the upper and lower surface; net photosynthetic efficiency of P. notoginseng showed that upper layer > middle layer > lower layer; there were significant correlation between soil temperature, stomatal conductance, intercellular CO2 concentration and photosynthetic rate were correlated with light intensity significantly; transpiration rates had notable correlation with light intensity and gas temperature. All above indicated that net photosynthesis rate of P. notoginseng was affected by light intensity directly, gas temperature and soil temperature indirectly. Inconclusion, stereoscopic cultivation of P. notoginseng was practicable in present study. The planting quality of P. notoginseng under stereoscopic cultivation could be improved by ameliorate the structure of seedbed to enhance the light intensity of middle and lower layer. Increase the thickness of the seedbed to decrease the temperature difference of soil. Further the management of ventilation facilities of greenhouse to control the gas temperature.
Carbon Dioxide
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metabolism
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Light
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Panax notoginseng
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growth & development
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metabolism
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Photosynthesis
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Soil
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Temperature
7.Expression of vascular endothelial growth factor in U937 foam cells and the inhibitory effect of drugs.
Peng-yuan YANG ; Yao-cheng RUI ; Li ZHANG ; Tie-jun LI ; Yan QIU ; Jie-song WANG ; Wei-dong ZHANG
Acta Pharmaceutica Sinica 2002;37(2):86-89
AIMTo study the expression of vascular endothelial growth factor (VEGF) in U937 foam cells and the inhibitory effect of salvianolic acid B and Ginkgo biloba extract in vitro.
METHODSU937 cells were incubated with 80 mg.L-1 oxidized low density lipoprotein (OX-LDL) for 48 h and a macrophage-derived foam cell model was established. The VEGF concentration in the media was determined by ELISA; the VEGF protein expression in cells was measured with immunohistochemistry; the VEGF mRNA level in cells was measured by in situ hybridization; the positive ratio detected by a morphometrical analysis system was used as the amount of the VEGF protein expression and the mRNA level.
RESULTSAfter U937 cells were incubated with OX-LDL, VEGF expression level increased greatly both in the cells and in the media. Salvianolic acid B and Ginkgo biloba extract were shown to remarkably inhibit the increase of VEGF. After treated with 10 micrograms/L-1 salvianolic acid B and Ginkgo biloba extract, the VEGF protein concentration in the media and positive ratio in the cells decreased compared with foam cells. After treated with 10 micrograms.L-1 salvianolic acid B and 100 micrograms.L-1 Ginkgo biloba extract, the VEGF mRNA level decreased measured by in situ hybridization.
CONCLUSIONA high VEGF expression level was determined in U937 foam cells. Salvianolic acid B and Ginkgo biloba extract were found to inhibit VEGF expression significantly in U937 foam cells in vitro.
Benzofurans ; pharmacology ; Foam Cells ; drug effects ; metabolism ; Gene Expression ; drug effects ; Ginkgo biloba ; chemistry ; Humans ; Plant Extracts ; isolation & purification ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; U937 Cells ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics
8.Effect of Curcuma aromatica on epithelial-myofibroblast transdifferentiation of NRK-52E cells induced by TGF-beta1.
Rui-Xuan WU ; Zhen-Fen HU ; Jin-Guo CHENG ; Wei-Xia HUANG ; Fei-Xia DONG
Chinese Journal of Integrated Traditional and Western Medicine 2013;33(7):972-977
OBJECTIVETo observe the antagonist effect of Curcuma Aromatica (CA) on renal tubular epithelial-myofibroblast transdifferentiation (EMT) induced by transforming growth factor-beta1 (TGF-beta1).
METHODSNormal renal tubular epithelial NRK-52E cells in vitro cultured were randomly divided into 6 groups, i.e., the normal control group (Group C), the TGF-beta1 induced model group (Group T), the low dose CA treated group (Group E1), the moderate dose CA treated group (Group E2), the high dose CA group (Group E3), and the Benazepril Hydrochloride Tablet treated group (Group Y). Except Group C, corresponding medication (with an action of 48 h) was administered to cells in the rest groups after they were induced by TGF-beta1 for 24 h. The morphological changes were observed by inverted phase contrast microscope. The distribution of beta-actin protein was detected by immunohistochemical assay. The mRNA expressions of alpha-smooth muscle actin (alpha-SMA) and E-cadherin (E-cad) were detected by real-time PCR. The concentration of fibronectin (FN) was detected by ELISA.
RESULTSAfter induced by TGF-beta1 for three days, hypertrophy and elongated cells in fusiform-shape occurred,with increased expressions of beta-actin protein in the cytoskeletal structure (P < 0.05), bundle fibrous structure occurred inside cytoplasm with significantly up-regulated intracellular alpha-SMA mRNA expressions (P < 0.05), E-cad mRNA expression decreased (P < 0.05), the FN content in the supernate increased (P < 0.05) in Group T. Compared with Group T, partial cells in Group E1, E2, and E3 showed fibrous changes, accompanied with decreased expression of beta-actin protein and FN concentration (P < 0.05). The expression of alpha-SMA mRNA increased and the expression E-cad mRNA decreased in Group E2 and E3 (both P < 0.05). But there was no statistical difference in the expression levels of E-cad and alpha-SMA mRNA (P > 0.05). Compared with Group E1, the expression of beta-actin protein and FN concentration decreased in Group E2 and E3 (P < 0.05). The expressions of alpha-SMA mRNA decreased and E-cad mRNA increased in Group E3 (P < 0.05). Compared with Group Y, the expression of beta-actin mRNA and FN concentration increased in Group E1 (P < 0.05); the expression of beta-actin mRNA increased in Group E3 (P < 0.05); the expression of E-cad mRNA decreased in Group E3 (P < 0.05). There was no statistical difference in the expression of alpha-SMA mRNA among Group E1, E2, and E3 (P > 0.05).
CONCLUSIONCA could inhibit the occurrence of TGF-beta1 induced EMT, which could be used as an effective drug for treating chronic renal insufficiency.
Animals ; Cell Transdifferentiation ; drug effects ; Cells, Cultured ; Curcuma ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; Kidney Tubules ; cytology ; Male ; Myofibroblasts ; drug effects ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism
9.Expression of recombinant human lysozyme-tachyplesin I (hLYZ-TP I) in Pichia pastoris and analysis of antibacterial activity.
Yu GAO ; Hong Lei ZHAO ; Xin FENG ; Rui Dong ZHAI ; Seng ZHU ; Chong Tao DU ; Chang Jiang SUN ; Lian Cheng LEI
Biomedical and Environmental Sciences 2013;26(4):319-322
Antimicrobial Cationic Peptides
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biosynthesis
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genetics
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DNA-Binding Proteins
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biosynthesis
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genetics
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Humans
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Microbial Sensitivity Tests
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Muramidase
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biosynthesis
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genetics
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Peptides, Cyclic
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biosynthesis
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genetics
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Pichia
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metabolism
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Recombinant Proteins
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biosynthesis
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genetics
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isolation & purification
10.Research development of the chemical material basis of Alisma orientalis and its toxicity.
Chun-fei WANG ; Xu-dong CHENG ; Jun-fei GU ; Jia-rui YUAN ; Bing-jie ZHAO ; Li ZHANG ; Juan CHEN ; Liang FENG ; Xiao-bin JIA
China Journal of Chinese Materia Medica 2015;40(5):840-846
Alisma orientalis is a traditional herb medicine commonly used in clinical. With the increasing report of its toxicity in clinical, the renal toxicity of Alisma orientalis has got gradually attention. This paper systematically reviews the research on the chemical material basis of Alisma orientalis including its chemical composition and toxicity of ingredients; and also declares its toxic ingredients and targets according to Network toxicology. Based on the controversy on renal toxicity of Alisma orientalis, we analyzed the possible reasons that may be associated with renal toxicity. It might be associated with the differences of the material basis composition and regulatory toxicology network, differences in employed processing technology, the metabolic function leading to accumulation of compounds, dosage and duration of the experiment and compatibility. The review provides possible reference and ideas for the quality control and rational use of Alisma orientalis.
Alisma
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chemistry
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toxicity
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Animals
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Drugs, Chinese Herbal
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chemistry
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toxicity
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Humans
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Molecular Structure