1.Anti-atherosclerotic Effect of Jiawei Zexie Tang in Rats
China Pharmacy 2005;0(24):-
OBJECTIVE: To study the antiatherosclerotic effect of Jiawei zexie tang(GWZXT) and the possible mechanisms.METHODS: 48 healthy male SD rats were randomized to 5 groups,i.e normal control,model group,simvastatin group,and high dosage and low dosage GWZXT groups.Artherosclerosis rat model was established by feeding high fat forage and the corresponding drugs were administered intragastrically.Changes in serum levels of APoAⅠ,ApoB,Zn,Cu,Ca and Mg and the ultrastructure were detected.RESULTS: In high dosage GWZXT-treated group compared with model group,the Zn,Ca and ApoB levels decreased significantly(P
2.Micropreparation of a Native PHGPx Protein From Radish Seedlings by Immunoaffinity Chromatography
Progress in Biochemistry and Biophysics 2005;32(8):794-799
Radish phospholipid hydroperoxide glutathione peroxidase (RsPHGPx) was identified as a mitochondrion-targeting PHGPx in previous work. To determine its cleavage site of the targeting peptide, the immunoaffinity chromatography (IAC) purification approach was carried out to isolate the native RsPHGPx protein.Polyclonal antibodies directed against recombinant RsPHGPx were raised in rabbit. Monospecific anti-RsPHGPx antibodies were isolated by means of affinity chromatography using the recombinant RsPHGPx as affinity ligand, and employed in assembling an IAC column. A single-step, highly specific and easy-to-use protocol was developed for purification of the active RsPHGPx protein through the assembled IAC column. Using this approach, a specific protein of the expected molecular size was obtained from the mitochondrial fraction of radish seedlings. Western blot analysis showed that it could be specifically recognized by anti-RsPHGPx antibodies, and an enzyme activity assay indicated that it exhibited significant PHGPx activity, suggesting that the purified protein should be the desired native RsPHGPx. These results will lead to clarification of the targeting peptide and the active mature protein of RsPHGPx and will be helpful to further probe the intracellular localization mechanism and biological fun ction of this plant PHGPx.
3.Radish Phospholipid Hydroperoxide Glutathione Peroxidase Gene Structure and Upstream Regulatory Sequence Analysis
Progress in Biochemistry and Biophysics 2005;32(7):649-656
A novel radish RsPHGPx cDNA, which encodes a functional phospholipid hydroperoxide glutathioneperoxidase (PHGPx) protein, was identified in the previous work. In the study genomic organization and the upstream regulatory sequence analysis of this gene was presented. Southern blot analysis showed that RsPHGPx gene existed in radish genome in manner of single copy. Moreover, a 3.3 kb genomic DNA fragment of RsPHGPx gene was isolated by combination of common PCR and genome-walking method. Sequence analysis on this genomic fragment demonstrated that RsPHGPx gene consists of seven exons separated by six introns, and suggested that a short 5'-flanking sequence immediately before the exon 1 should be the putative RsPHGPx promoter region, which is proceeded by the upstream neighboring biotin synthase gene. Cis-acting elements search showed that the putative promoter contains elements responsive to hormones (eg. E-Box and W-Box), abiotic stresses (eg. MYB and MYC binding sites), and light (Box Ⅱ and Ⅰ-Box), etc. Northern blot analysis indicated that the expression of RsPHGPx was subjected to up-regulation of chilling and down-regulation of ABA and successive illumination (in etiolated seedlings), implying the regulatory roles of some predicted elements. However the up-regulation effect of herbicide paraquat, which can induce oxidative stress, suggested the presence of some unknown elements in the promoter region. This is the first report on gene structure and upstream regulatory sequence analysis in reported plant PHGPx genes, which will be a prerequisite to understand regulatory mechanism of PHGPx gene expression in plants.
4.Tissue and Induction Expression Profiles of Rice Phospholipid Hydroperoxide Glutathione Peroxidase at Protein Level
Tian LI ; Xiaodong YANG ; Jinyuan LIU
Progress in Biochemistry and Biophysics 2009;36(1):77-82
Proteins arc the major molecules performing life activities, and their spatial and temporal expression profiles in organisms are very important for understanding their accurate functions. Phospholipid hydroperoxide glutathione pcroxidase (PHGPx) is a unique antioxidant enzyme that directly reduces lipid hydroperoxides in biomembranes and plays a vital role in defense against mernhrane peroxidation damage. The protein expression profiles of rice PHGPx (OsPHGPx) were investigated in different rice tissues and under various stress treatments by using Western blot analysis. The results showed that in mature rice plants, OsPHGPx was mainly distributed in leaves, especially flag leaves, and in rice seedlings, OsPHGPx was detected in shoots and leaves. Moreover, OsPHGPx expression in rice seedlings could be markedly induced by H2O2 and NaCI, but weakly influenced by several plant hormones. Time- and dose-dependent effects were observed in both H2O2 and NaCI treatments, and the strongest induction was observed when rice seedlings were treated with 0.5 mmol/L H2O2 for 12 h or 500 mmol/L NaCI for 24 h. Additionally, dimethylthiourea, a H2O2 trap, inhibited H2O2-enhanced expression of OsPHGPx, but did not impair the enhanced effect of NaCI, implying that NaCl-induced OsPHGPx expression was not mediated by H2O2. These results will contribute greatly to further study the exact physiological function of OsPHGPx in rice.
5.Interventional therapy for congenital urinary obstruction in children
Zenhui QIN ; Sui HHANG ; Fan LIU ; Jinyuan YANG
Journal of Interventional Radiology 1994;0(04):-
Objective To investigate the interventional therapy in children's congenital urinary obstruction and its efficacy. Methods Thirty-three children with congenital obstruction of ureteropelvic junction were treated through percutaneous dilation and/or stent placement, and 42 cases with posterior urethral valves were treated through trans-urethra dilation. Results Thirty-three cases with upper urinary obstruction were improved with symptoms disappeared and stable efficacy on long-term follow-up of 1-7 years. Another 2 cases with the upper urethral obstruction had not been relieved of symptoms and resorted to surgical operation. For patients with posterior urethral valves, the lower urethral obstruction was totally got rid of after interventional therapy with stable efficacy on long-term follow-up of 1-10 years. Conclusions Interventional therapy is safe, micro-invasive and efficient in treating congenital urinary obstruction with stable efficacy on long-term follow-up.
6.Treatment of children spleenomegaly by partial splenic embolization
Fan LIU ; Zenghui QIN ; Liangbo XU ; Sui HUANG ; Jinyuan YANG
Journal of Interventional Radiology 1994;0(04):-
Objective To investigate the safety treatment of partially embolizing spleenomegaly in children. Methods Forty two children aged 1-15 with spleenomegaly were treated through staged partial splenic embolization (PSE). The first embolized scope of spleen was 30%-40% with the second being 30%-40% at 1 or 2 months later in order to achieve the goal of getting rid of hypersplenia and improving the splenic function. Results The adverse effects of splenic embolisation were slight with short duration of fever and stomachache and with efficient control of hypersplenia and its correlative basic diseases except one case of splenic abscess.Conclusions Spleenomegaly in Children can be more safely and more efficiently cured through staged PSE.
7.Downregulation of heat shock protein B8 protects retinal ganglion cell after optic nerve injury in mice
Feijia XIE ; Tao HE ; Ning YANG ; Jiayi YANG ; Dihao HUA ; Jinyuan LUO ; Zongyuan LI ; Yiqiao XING
Chinese Journal of Ocular Fundus Diseases 2021;37(4):298-306
Objective:To investigate the effect of heat shock protein B8 (HspB8) downregulation on retinal ganglion cell (RGC) and retinal function in the mice model of optic nerve injury (ONC).Methods:Adeno-Associated Virus (AAV) 2 AAV2-shHspB8-GFP was constructed to knockdown HspB8. 66 adult male C57/BL6 mice were randomly divided into the control group, the ONC group, the AAV2-shHspB8 group, the ONC+AAV2-shHspB8 group, and the ONC+AAV2-GFP group. There were 10, 20, 16, 10 and 10 mice respectively, and both eyes were used as experimental eyes. Western blot was used to evaluate the expression of HspB8 on day 3 and 7 after ONC. By GFP immunofluorescence staining, the efficacy of AAV2-shHspB8-GFP transfer was accessed. Moreover, it was possible to identify functional and RGC survival differences between groups by optomotor response (OMR), dark adapted full-field flash electroretinogram (ff-ERG), oscillatory potentials (OPs), photopic negative response (PhNR) and retinal flat-mount RGC counting 5 days after ONC. Comparisons between two groups were made using Mann-Whitney U test, unpaired t-test, unpaired t-test with Welch’s correction, one-way ANOVA, and Bonferroni t test. Results:Compared with the control group, the expression of HSPB8 protein in the retina of mice in ONC3 group was significantly increased, and the difference was statistically significant ( F=43.63, P<0.01). Compared with the control group, the ONC group showed obviously lower visual acuity ( P<0.01), lower a-wave, b-wave, OPs, PhNR amplitude, longer b-wave latency ( P<0.05), and the survival rates of RGC in ONC3 group, ONC5 group and ONC7 group decreased in a time-dependent manner( F=384.90, P<0.01). Transfection of AAV2 efficiency was highest on 4 weeks after IVT. Besides, there was no significant differences between the control group and the AAV2-shHspB8 group on visual acuity, ff-ERG, OPs, PhNR and RGC survival ( P>0.05). In comparison of the control group, we found that RGC survival of the ONC5+AAV2-shHspB8 group was significantly elevated ( F=10.62, P<0.01). Conclusions:Expression of HspB8 on the retina can be induced by ONC. The investigation of RGC counting, visual acuity, and ff-ERG revealed that optic nerve injury destructed functionality of mice retina and resulted to RGC death ultimately. The Most crucial finding of this research is that HspB8 knockdown had a neuroprotective effect in RGC after ONC.
8.Effects of N-acetylcysteine on Alcoholism in Mice
Yongping HUANG ; Shiwen ZHOU ; Ying XU ; Jianlin TANG ; Lian WANG ; Jinyuan ZHANG ; Xue YANG
China Pharmacy 2001;0(07):-
OBJECTIVE:To investigate the effects of N-acetylcysteine(NAC)on the concentration of ethanol in serum and the activities of alcohol dehydrogenase in liver and stomach tissue in alcoholic mice.METHODS:All mice were given p.o white spirit30min after p.o saline or5%NAC solution,and the durations from the disappear of righting reflex to recovery in mice were recorded and the number of dead mice in24hours were countered;With the same procedure for six days,the mice were killed by withdrawaling blood from orbit,and taken out the liver and stomach immediately,the concentrations of ethanol in serum and the activities of alcohol dehydrogenase in liver and stomach were measured by chromatometry and colorimetric method respectively.RESULTS:In drunk test,5%NAC solution significantly postponed the time from p.o white spirit to the disappear of righting reflex(P
9.Vector construction and expression of soluble mPDL1-hIgGFc and its effect on the proliferation and apoptosis of cells in vitro
Jing YANG ; Wenjun LIAO ; Guohua WANG ; Fengrong HE ; Huifen ZHU ; Hong DAI ; Wei ZHOU ; Xiongwen WU ; Jinyuan ZHANG ; Guanxin SHEN
Chinese Journal of Microbiology and Immunology 2008;28(9):795-798
Objective To construct vector expressing soluble mPDL1-hIgGFc and study its effect on the proliferation and apeptosis of cells in vitro. Methods The extrncellular domain of mPDL1 gene was amplified from pmPDL1 vector by PCR and inserted into phIgGFc vector. The recombinant pmPDL1-hIgGFc was transfected into CHO cells by LipofectAMINETM2000, and the transfected cells were named as CHOp. The expression of mPDL1-hIgGFc in the culture supernatants of CHOp was assayed by ELISA and Western blot. The effects of CHOp culture supernatants on mixed lymphocyte culture(MLC) was analysed by Flowm-etry. Results The extracellular domain of mPDL1 gene were obtained from PCR. DNA sequencing and the identification of digestion by HindⅢ and KpnⅠ indicated the recombinant plasmid pmPDL1-hIgGFc was suc-cessfully constructed. ELISA and Western blot analysis proved that the CHOp could express mPDL1-hIgG-Fc. CHOp culture supernatants could inhibit lymphocyte proliferation and induce the apoptosis of the activa-ted T cells in MLC in vitro in a dose-dependent manner. Conclusion The mPDL1-hIgGFc protein could in-hibit lymphocyte proliferation and induce the apoptosis of the activated T cells.
10.Effects of Krüppel-like factor 7 on the survival of retinal ganglion cells and electroretinogram after retinal ischemia-reperfusion injury
Zongyuan LI ; Ning YANG ; Jinyuan LUO ; Jiayi YANG ; Tao HE ; Yiqiao XING
Chinese Journal of Ocular Fundus Diseases 2020;36(11):846-852
Objective:To investigate the effects of Krüppel-like factor 7 (KLF7) on the survival of retinal ganglion cells (RGCs) and electroretinogram (ERG) after retinal ischemia-reperfusion (RIR) injury in mice.Methods:A total of 126 male C57BL/6J mice were randomly divided into normal group, RIR group, normal-KLF7 group, normal-green fluorescent protein (GFP) group, RIR-KLF7 group and RIR-GFP group. At the age of 8 weeks, mice of normal-KLF7 group and RIR-KLF7 group were intravitreally injected 1ul of 1.0×10 12 vg/ml adeno-associated virus overexpressing KLF7 (AAV2-KLF7-GFP). Mice of normal-GFP group and RIR-GFP group were injected adeno-associated virus of AAV2-GFP with the same titer. At the age of 11 weeks, RIR injury was induced in mice of RIR group, RIR-KLF7 group and RIR-GFP group, and intraocular pressure was measured. Retinal cryosections were used to access the efficacy of virus transfection 4 weeks after AAV2-KLF7-GFP transfer. 7 days after RIR injury, RGCs' survival rate was observed and quantified by immunofluorescent staining. ERG was performed to observe the differences in amplitudes and incubation period of scotopic ERG a-, b-wave, oscillatory potentials (Ops), photopic negative responses (PhNR). Optomotor response was performed to observe the differences of visual acuity. Expression of KLF7 was detected by western blot 4 weeks after AAV2-KLF7-GFP transfer. Results:Compared with normal group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR group were decreased, and the differences were statistically significant ( t=12.860, 7.157, 5.735, 8.953, 4.744, 9.887; P<0.05). With the increase of light intensity, the amplitudes of scotopic ERG a- and b-wave were gradually increased while the incubation period was gradually shortened. Compared with RIR group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR-KLF7 group were increased, and the differences were statistically significant ( t=6.350, 3.253, 3.695, 5.825, 5.325, 4.591; P<0.05). Protein level of KLF7 was up-regulated in normal-KLF7 group than those in normal group, and the difference was statistically significant ( t=4.105, P<0.01). Conclusion:Overexpression of KLF7 can improve RGCs’ survival rates and preserve the electrophysiological function.