1.Dynamic changes of FasL protein expression in retinal tissue with aging in retinal degeneration mice
Chinese Journal of Experimental Ophthalmology 2017;35(1):17-21
Background Retinal transplantation is a new approach to the treatment of retinal degeneration diseases,and how to avoid or reduce the immune rejection after transplantation is a problem.In experimental studies on retinal transplantation,C57BL/6 mice are often used as donors and rd mice serve as recipients.Studies showed that Fas ligand (FasL) protein induces the apoptosis of Fas+ inflammatory cells by FasL/Fas signal pathway,speculating that FasL protein is associated with immune rejection after transplantation.Objective The aim of this study was to investigate FasL protein expression change in retinas of C57BL/6 mice and rd mice with aging and reveal the immune characteristics of the mouse retinas.Methods The frozen sections of eyeball from C57BL/6 mice and rd mice at the age of postnatal (PN)-0 week,PN-1 week,PN-2 week,PN-3 week and PN-4 week were prepared.The expression of FasL protein in the mouse retinas was examined by immnofluorescense technique.Images were acquired by fluorescence microscope and analyzed semi-quantitively by software from laser scanning confocal microscope as the fluorescence intensity (FI).The results were compared among different strains of mice.Results The retina developed imperfectly in PN-1 week C57BL/6 mice and FasL protein was positively expressed in the whole retina.In PN-2,3 and 4 week C57BL/6 mice,retinas finished the development with 10 layers,and retinal pigment epithelium (RPE),inner segment (IS),outer limiting membrane (OLM),outer plexiform layer (OPL),inner nuclear layer (INL),inner plexiform layer (IPL) and ganglion cell layer (GCL).Retinal structure and expression of FasL protein in the whole retina of rd mice in the PN-1 week were similar with C57BL/6 mice,however,ONL cells of rd mice were evidently decreased with aging.The RPE,OPL,INL,IPL and GCL expressed the FasL protein in PN-2,PN-3 and PN-4 week rd mice.The mean FI of FasL protein in the RPE layer was 184.199±16.747,186.797±7.904 and 184.319± 18.795 in rd mice of PN-2 week,PN-3 week and PN-4 week,which were significantly higher than 160.402±22.851,160.995 ±22.799 and 105.787 ± 17.676 in C57BL/6 mice (t =-3.360,P =0.002;t'=-4.277,P =0.000;t =-12.175,P=0.000).There were not significant differences in the mean FI of FasL protein in IPL between C57BL/6 mice and rd mice at ages of PN-2 week,PN-3 week and PN-4 week (all at P>0.05).Conclusions The cells of retinal ONL are gradually decreased with the development of rd mice,and FasL protein expression intensity in RPE is evidently enhanced in rd mice compared with C57BL/6 mice.
2.Clinical Research on Elderly Hypertension of Kidney Deficiency and Blood Stasis Syndrome Intervened with Bushen Huoxue Decoction
Chinese Journal of Information on Traditional Chinese Medicine 2015;(2):25-27
Objective To observe the clinical efficacy of Bushen Huoxue Decoction on elderly hypertension of kidney deficiency and blood stasis syndrome. Methods Totally 106 patients with elderly hypertension of kidney deficiency and blood stasis syndrome were divided into control group (53 cases) and treatment group (53 cases), according to random number table method. The control group was treated with routine western medicine, while the treatment group was given Bushen Huoxue Decoction along with western medicine treatment for 8 weeks. The changes of TCM syndrome, blood pressure, TG, TC, LDL-C, HDL-C, high sensitivity C reactive protein (hs-CRP), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) before and after treatment of the two groups were observed. Results The total effective rate of anti-hypertension was 94.33%(50/53) in the treatment group and was 77.35% (41/53) in the control group. The total effective rate of clinical symptoms was 96.22%(51/53) in the treatment group and was 79.24%(42/53) in the control group. After treatment, TG, TC, LDL-C, HDL-C, hs-CRP, IL-6, and TNF-α were improved significantly in the treatment group, with statistical significance compared with the control group (P<0.05). Conclusion Bushen Huoxue Decoction combined with western medicine achieves the good efficacy on elderly hypertension of kidney deficiency and blood stasis syndrome.
3.Ultrasonography of jugular vein in assessing hypovolemia in healthy blood donor
Dan TIAN ; Wenqiang LI ; Jie WEI
Chinese Journal of Postgraduates of Medicine 2015;38(z1):74-76
Objective To assess volume state precisely and rapidly by ultrasonography of internal jugular vein (IJV) in healthy blood donor.Methods The values of the sonographic IJV collapse index and corrected IJV longitudinal length (cIJVLL) of 46 healthy blood donors were compared before and after blood donation.The correlations between IJV collapse index and cIJV LL were analyzed.Results The value of cIJV LLs before and after blood donation were significantly difference (6.56 ± 0.32 vs.6.11 ± 0.41,P < 0.01).IJV collapse index before blood donation was not differently significant after blood donation (33.12 ± 2.21 vs.39.01 ± 3.83,P> 0.05).There was correlation between the value of cIJV LLs before and after blood donation (r =0.81).The value of IJV collapse index before and after blood donation,as well as cIJVLL was not well correlated (r =0.24,r =0.13,respectively).Conclusion The IJV collapse index is not a useful parameter for evaluation of hypovolemia,cIJV LL is more valuable marker for the detection of blood loss in emergency.
4.Research Progress of Taking Key Enzyme in Metabolism as Selectable Marker in Transgenic Plant
Hui-Yong JIA ; Jia TIAN ; Jie LI ;
China Biotechnology 2006;0(03):-
In plant genetic engineering, selectable marker is needed to distinguish transformant. As the commercialization of transgenic plants, people are more and more paying close attention to their safety, among which mainly refers to the safety of selectable marker. In order to increase the safety of transgenic plants, biologists began to search for biosafe selectable marker.Current research progress of taking key enzyme in metabolic pathways were overviewed,which include glycometabolism, amino acid metabolism, hormone metabolism ,nucleotide metabolism and protein metabolism et cetera, as selectable marker in transgenic plants.
5.Effect of high-flux hemodialysis on advanced oxidation protein products in maintenance hemodialysis patients
Chunhong LI ; Tian XIA ; Jie TIAN ; Hongling WANG
Chinese Journal of Geriatrics 2013;32(12):1308-1311
Objective To investigate the effect of high-flux hemodialysis (HFHD) on advanced oxidation protein products (AOPPs) in maintenance hemodialysis (MHD) patients.Methods 40 MHD patients (MHD group),50 non-dialytic chronic kidney disease (CKD) patients (CKD group) and 31 healthy subjects undergoing health examination survey received AOPPs detection.After lowflux hemodialysis (LFHD) for 3 months,HFHD was performed in MHD group for 3 months.Plasma AOPPs and other biochemical indicators were tested at the end of 0,3 and 6 months.AOPPs levels were detected in HFHD and LFHD dialysates.Results Plasma AOPPs level was higher in MHD group than in CKD and control groups,and plasma AOPPs level was higher in CKD group than in control group.Compared with before LFHD in MHD group,AOPPs and triglyceride (TG) levels had no significant differences after LFHD for 3 months(t=0.831,-0.191,both P>0.05),while plasma cholesterol (CHO) and β2 MG levels were increased in MHD group after LFHD for 3 months(t=-2.697,10.39,P<0.05 or 0.001).There were significant differences in plasma AOPPs,TG,β2-MG and CHO levels before versus after HFHD for 3 months (t=4.292,2.271,8.297 and 2.295,P <0.001 or 0.05).No AOPPs were detected in HFHD and LFHD dialysates.Conclusions Plasma AOPPs levels are increased in CKD patients.LFHD can not effectively eliminate AOPPs,while long term HFHD can decrease the plasma levels of AOPPs,TG and CHO,and reduce β2-MG accumulation.
6.Platelet-rich plasma effects on grainy fat transplantation
Feng LI ; Kun LI ; Jie LI ; Weidong TIAN
Chinese Journal of Tissue Engineering Research 2013;(31):5645-5651
BACKGROUND:The higher long-term absorption rate greatly influence the widely application of fat transplantation. Platelet-rich plasma contains a high concentration of growth factors, which benefits to the tissue healing and regeneration. OBJECTIVE:To observe the effects of platelet-rich plasma on grainy fat transplantation and to investigate the mechanisms preliminarily. METHODS:Ten 6-week-old nude mice were prepared. The right or left dorsal subcutaneous tissues were randomly selected as the platelet-rich plasma group (0.5 mL fat granule+0.1 mL platelet-rich plasma), and the contralateral side was regarded as the control group (0.5 mL fat granule+0.1 mL phosphate-buffered saline). At 10 and 90 days after implantation, five nude mice were selected from each group, and then the mice were sacrificed to obtain the grafts in each group for general appearance observation, volume determination and histological detection. Furthermore, we isolated adipose-derived stem cells from human subcutaneous fat tissue during the in vitro experiment. Cel counting kit-8 and real-time PCR were used to evaluate the influence of platelet-rich plasma on adipose-derived stem cel proliferation and adipogenic differentiation in vitro, respectively. RESULTS AND CONCLUSION:Comparison of the grafts obtained at 10 and 90 days after implantation, the residual volume in the platelet-rich plasma group was significantly larger than that in the control group (P<0.05), Moreover, more normal adipocytes and capil ary formation were observed in the platelet-rich plasma group (P<0.05). For in vitro experiment, platelet-rich plasma could significantly improve adipose-derived stem cel proliferation, and the expressions of adipogenic-related genes were up-regulated in platelet-rich plasma-induced adipose-derived stem cells. Al results demonstrate that platelet-rich plasma can improve the survival of fat grafts,which might be closely related to that platelet-rich plasma can promote the proliferation and adipogenic differentiation of adipose-derived stem cells and the revascularization in grafted fat tissue.
7.Determination of Gallic Acid and Kaempferol-3-O-Rutinoside Contents inNymphaea Candida Presl. by HPLC
Jie LI ; Xuejia ZHANG ; Keao LI ; Zhihong JI ; Shuge TIAN
Chinese Journal of Information on Traditional Chinese Medicine 2014;(8):92-94
Objective To establish an HPLC method for the content determination of gallic acid and kaempferol-3-O-rutinoside inNymphaea candida Presl..MethodsThe separation was carried out on a Wondasil C18 column (250 mm×4.6 mm, 5μm). The mobile phase was methanol-0.1% acetic acid solution, with gradient elution;the flow rate was 1.0 mL/min;the detection wavelength was set at 270 nm;the column temperature was 30℃.Results The linear ranges of gallic acid and kaempferol-3-O-rutinoside were 0.065-0.585μg (r=0.999 1), 0.133-1.197μg (r=0.999 5), respectively. The average recoveries of gallic acid and kaempferol-3-O-rutinoside were 102.71%, 102.08%, with RSD of 1.97%, 0.46%, respectively.Conclusion This method was rapid, accurate, and reliable. It can be used for the determination of allic acid and kaempferol-3-O-rutinoside in Nymphaea candidaPresl..
8.Screening of plasmids targeted to GCN5 and detection of their effects on MSCs differentiation
Li LI ; Jing ZHU ; Chuan FENG ; Jie TIAN
Basic & Clinical Medicine 2006;0(06):-
Objective To screen the valid plasmid targeted toGCN5 among the constructed recombinant plasmids;and to explore the effects of histone acetylizad modification in regulating MSCs differentiation.Methods Exstract the constructed plasmids and transfect them into MSCs induced by 5-aza.For 24 h,observe MSCs;detect the transfect efficiency by flow cytometry;detect expression of protein GCN5 by Western-blot;detect the HAT activity by ELISA.Results Transfect efficiency was more than 20%;Expression of protein GCN5 and HAT activity had no difference in group ZJ1 and ZJ2,but had a significant difference to that in group ZJ3.HAT activity of experiment group was significantly lower than that of control groups.Conclusion The inhibition state of histone acetylation results in plasmid ZJ3,it can inhibit the differentiation process of MSCs.The results provide data for the clinical application of MSCs transplantation.
9.Effect of Momordicin on Expression of Tumor Necrosis Factor-? in Balb/c Mice with Viral Myocarditis Caused by Coxsackievirus B_3
hong, TIAN ; xiao-ming, LI ; shuang-jie, LI
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To observe the effect of momordicin on tumor necrosis factor-?(TNF-?) level,mRNA transcription,and protein expression in myocardium of viral myocarditis caused by coxsackievirus B3(CVB3),and explore its therapeutic mechanism on viral myocarditis in Balb/c mice.Methods Fifty Balb/c mice were randomly divided into 3 groups as follows:momordicin treatment group(20 cases),vehicle control group(20 cases) and normal control group(n=10).Mice in the vehicle control group and the momordicin treatment group were intraperitoneally inoculated with CVB3,as for the nomal control group,equal amount of culture fluid was given instead.Momordicin[25 mg/(kg?d)] was administered intraperitoneally daily from day 0 to 6.Myocardial histopathology,cardiac TNF-? antigen,protein and mRNA expression were detected on day 15 after CVB3 inoculation,respectively.Results As compared with model group,in mice treated with momordicin,the histological myocardial lesion was significantly reduced [(3.26 ?0.84) vs(1.56?0.48),t=3.90 P
10.Therapeutic Effect of Astragaloside on Chronic Coxsackievirus B_3 Myocarditis in Balb/c Mice
xiao-ming, LI ; hong, TIAN ; shuang-jie, LI
Journal of Applied Clinical Pediatrics 2006;0(18):-
Objective To observe the therapeutic effect of astragaloside on chronic coxsackievirus B3(CVB3)myocarditis in Balb/c mice.Methods Eighty Balb/c mice were randomly divided into 3 groups:astragaloside treatment group(n=30),model of viral myocarditis group(model group)(n=30),and control group(n=20).Mice in the model group and the astragaloside treatment group were monthly intraperitoneally inoculated with CVB3,but equal amount of culture fluid was given instead in control group.The model group and control group were fed with drinking water,astragaloside treatment group were fed with drinking water containing astragaloside at concentration of 300 mg/L for 3 months.Survival rates were determined,myocardial histopathology,collagen volume fraction(CVF) and apoptosis of heart tissue,and CVB3 RNA levels were detected on 3 months later respectively by semiquantitative RT-PCR.Results Compared with model group,in astragaloside treated group,the survival rate on 3 months was significantly improved(59.7% vs 76.7%,?2=4.26 P