4.Effects of glycyrrhizic acid and solid dispersions of curcumin on anti-oxidation and components of lung lavage fluid of SiO2 exposed rats.
Gang HAN ; Lin-Qi YAN ; Zhi-Yuan CHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2010;28(4):289-290
Animals
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Bronchoalveolar Lavage Fluid
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chemistry
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Curcumin
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pharmacology
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Glycyrrhizic Acid
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pharmacology
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Lung
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drug effects
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metabolism
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Male
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Rats
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Rats, Wistar
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Silicon Dioxide
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toxicity
5.One case of self-reinfusion of surfactant in whole lung lavage.
Jing-Hui HUANG ; Zhi-Yuan CHEN ; Gang CHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2010;28(12):948-949
Adult
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Bronchoalveolar Lavage
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methods
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Humans
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Male
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Pulmonary Surfactants
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therapeutic use
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Silicosis
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therapy
6.Purification and Characterization of Glucagon-Like Peptide-1 and Human Serum Albumin Fusion Protein Expressed in Pichia pastoris
Jia-Qi CHEN ; Zhi-Hui GAO ; Yuan-Yuan ZHU ; Wen-Bo YANG ; Gang BAI ;
Microbiology 1992;0(05):-
A fusion protein of glucagons-like peptide-1 and human serum albumin (GLP-1/HSA) was expressed and secreted into the fermentation broth with recombinant Pichia pastoris. The productivity of expressed GLP-1/HSA could reach 63.6mg/L in 10L fermentor. After concentrated with hollow-fiber ultrafiltration membrane, GLP-1/HSA was purified from fermentation broth by hydrophobic chromatography, negative ion exchange chromatography and gel filtration chromatography in turn. The HPLC analysis showed that the purified GLP-1/HSA had an overall purity of 95.8%. Furthermore, the analysis of in vivo activity indicated that GLP-1/HSA had the bioactivity of native GLP-1, and could significantly reduce blood glucose level 4h after intraperitoneal administration. It was concluded that a great deal of GLP-1/HSA with higher purity could be harvested by Pichia pastoris expression system and the established purification methods. Preliminary studies show a new potential for developing the long-acting GLP-1 analogs for clinical applications.
7.Adrenal tuberculosis:contrast-enhanced CT features with clinical correlation
Zhi-Gang YANG ; Ying-Kun GUO ; Yuan LI ; Peng-Qiu MIN ; Jian-Qun YU ;
Chinese Journal of Radiology 1999;0(10):-
Objective To investigate the correlation between specific features on contrast-enhanced CT and its clinical findings of adrenal tuberculosis so as to improve the diagnostic accuracy of the disease. Methods Contrast-enhanced CT features in 30 patients with documented adrenal tuberculosis were retrospectively evaluated blindly for the features of location,size,morphology,attenuation and enhancement patterns on CT images,and compared with clinical and pathological materials.Results The common CT manifestations were as follows:enlargement of the adrenal glands in all 30 cases(bilateral 90%,mfilateral 10%)including mass-like enlargement in 13 cases and enlargement but the contours preserved in 17 cases, heterogeneity(28 cases,93.3%),calcification(17 cases,56.7%),and low attenuation in the center with peripheral enhancement(16 cases,53.3%)of the lesions.After antituberculosis chemotherapy, 5 cases of enlarged adrenal glands decreased in size or returned to normal size and configuration,with disappearance of the central low attenuation and new appearance of dot-like calcification in 2 cases.Cochran Armitage trend test showed there was an increasing tendency of calcification rate with clinical duration(X~2= 7.47,P
8.Inhibition of subretinal co-transfection of Rb94 and wild-type p53 gene on retinoblastoma by ultrasound microbubble in nude mouse
Rui-Qi, GAO ; Xi-Yuan, ZHOU ; Ying-Xue, YANG ; Zhi-Gang, WANG
Chinese Journal of Experimental Ophthalmology 2012;(10):908-913
Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80% (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P<0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.
9.Influence of hypoxia on cell proliferation and expression of p21 in rhesus retinal vascular endothelial cells
Hua-bin, ZHENG ; Jin-dong, HAN ; Zhi-gang, YUAN ; Hua, YAN
Chinese Journal of Experimental Ophthalmology 2012;(11):972-975
Background Retinal neovascularization disease is a common cause of blinding.Retinal neovascularization is related to enhancing proliferation of vascular endothelial cells.So how to inhibit proliferation of vascular endothelial cells is a hot burning issue.p21 is known to be involved in the regulation of cell cycle and therefore inhibit the cell proliferation.However,the relationship of p21 and the proliferation of vascular endothelial cells in retinal neovascularization disease is for further study.Objective The aim of this experiment was to study the proliferation of rhesus retinal vascular endothelial cells(RF/6A) and expression of p21 in RF/6A cells under the hypoxia condition,and discuss their association.Methods The RF/6A cells were cultured and passaged in vitro,then they were randomly divided into normoxia culture group(5% CO2 +95% O2) and hypoxia for 1 hour,3,6,12 hours group(1% 02+5% CO2 +94% N2).Flow cytometer(FCM) was used to check the distribution of RF/6A cell cycle in the normoxia culture group and hypoxia for 1 hour,3,6,12 hours groups.MTT assay was used to detect and compare the cell proliferation(A570)among the various groups.The expression of p21 in the cells was analyzed by Western blot.Results FCM showed that the cells proportion of G0/G1 stage was reduced initially and then increased afterward in hypoxia for 1 hour and 3,6,12 hours groups,showing a significant difference among 5 groups (F =20.083,P =0.000),and the cells proportion of S stage and G2/M stage were increased firstly and then declined in different hypoxia groups with statistical significances (F =7.861,P =0.001 ; F =10.305,P =0.003).Compared with normoxia culture group,cells proportion of G0/G1 stage was declined and that of S stage and G2/M stage were raised after hypoxia culture,showing statistically signifcant differences(P<0.05).MTT showed that cell multiplication capacity(A570 value)strengthened firstly and then weakened in hypoxia groups with time prolongation,showing a significant difference among all the groups(F=7.768,P=0.001),and A570 value in hypoxia for 3 hours and 6 hours groups (0.315± 0.062,0.365 ± 0.064) was significantly higher than that of the normoxia group (0.205 ± 0.063),respectively(P<0.05).Western blot showed that the expression of p21 in the cells down-regulated at the beginning and then up-regulated with the increase of hypoxia time,and there was statistical significance (F =16.738,P=0.000).The p21 relative levels in different hypoxia groups were reduced in comparison with the normoxia group,showing statistical signifcances(P<0.05).Conclusions Short-term hypoxia could reduce the expression of the p21 in RF/6A and induce cell proliferation initially,then p21 increases and cell proliferation is inhibited with the prolongation of hypoxia time.
10.The effects of nitric oxide synthase and its antagonist on alkali burn-induced corneal neovascularization
Gao-qin, LIU ; Yuan, CHEN ; Lei, CHEN ; Yan-hui, XIAO ; Zhi-gang, CHEN ; Pei-rong, LU
Chinese Journal of Experimental Ophthalmology 2013;31(10):908-913
Background Though nitric oxide (NO) and NO synthase (NOS) have a critical role in angiogenesis,their effects on corneal neovascularization (CNV) and mechanism need to be further explored.Objective The aim of this study was to explore the effects of NOS and its antagonist,Nw-nitro-L-arginine methyl ester (L-NAME) on experimental CNV in mice,and investigate the influence of NOS and L-NAME on the tube formation of human retinal endothelial cells (RECs) in vitro.Methods The CNV models were established in the left eyes of 36 male BALB/c mice aged 7-8 weeks by application of the filter paper with NaOH in the center of corneas.The mice were randomized into two groups.L-NAME of 10 g/L (0.5 ml) was intraperitoneally injected 1 week before induction of CNV three times a week for three weeks in the mice of the L-NAME injection,and PBS was used in the same way in the control group.CNV was examined under the slit lamp biomicroscope 2,4,7,14 days after NaOH burn.The expression of CD31 in the CNV was assayed to calculate the ratio of CNV area and total corneal area using whole mount technique.The expression of NOS mRNA in the corneal tissue was detected by reverse transcriptase polymerase chain reaction (PCR),and VEGF expression in the human RECs was assayed by Western blot.The vessel formation number of cultured human RECs with or without L-NAME was performed by matrigel in vitro.Grouped t test was used to compare the differences of the parameters between the two groups.Results CNV developed and peaked 2 weeks after the application of NaOH on the mice corneas,and the CNV was obviously less in the L-NAME group compared with the control group.The expression of NOS mRNA in the corneas (NOS mRNA/ GAPDH mRNA)was significantly lower in the L-NAME group than that of the control group 2,4,7 days after CNV induction (t =19.481,t=22.059,t=10.961,all at P<0.01).The ratio of the CD31 positive area in whole corneal area was 0.59± 0.01 in the L-NAME group,and that of the control group was 0.78±0.10,showing a significant difference between the two groups (t =3.078,P<0.05).Western blot assay showed that the relative expression of VEGF protein in human RECs was declined in the L-NAME group compared with the control group 0,2,4,7 days,with statistically significant differences in 4 days and 7 days after NaOH burn(t=7.696,t=17.953,both at P<0.01).The number of vessel network was 46.33±1.86 in the L-NAME group and 64.00±4.51 in the control group,with a significant difference between them (t =3.623,P<0.05).Conclusions NOS participated in the pathogenesis and aggravation of CNV induced by NaOH.L-NAME arrests CNV formation and human RECs tube formation through down regulating the VEGF expression and NOS activity.