2.A feasibility study of recombinant adeno-associated virus(rAAV)as a vector for transferring a target gene to retina
Jian-Ming, WANG ; Ya-Zhi, FAN ; Na, HUI ; Lei, XIONG ; Hai-Xiao, FENG ; Nai-Xue, SUN
International Eye Science 2008;8(9):1740-1742
AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.
3.Development of three dimensional facial measurement system based on structured light projection
Yao-yang, XIONG ; Xiao-bo, CHEN ; Jian, SUN ; Fu-qiang, ZHANG ; Jun-tong, XI
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(7):837-841
Objective To construct a system for three dimensional face scanning and measurement. Methods The measurement system was based on the principle of triangulation and the combination of gray-code and phase-shift structured light projection. The system software was developed for Windows XP with the aid of tools such as Visual C++ and Hoops. Results A three dimensional measurement system based on structured light projection was developed. The system hardware was composed of fringe projection unit, image gathering unit, system control unit and mechanical appearance, and the system software was composed of point cloud display and editing module. The lamp house of the system was 12V, the working distance was 900 mm, the scanning time was 5.5 s and the scanning field was 500 mm×400 mm. Conclusion The three dimensional measurement system based on structured light projection is a refined machine with safe light to eyes, and the accuracy and scanning speed are suitable to face scanning.
4.Activity and expression of aldehyde dehydrogenases in the ocular tissue of diabetic rats
Ming-ling, WANG ; Shu-fang, HU ; Jian-xiong, CAO ; An-an, WANG ; Tian-lin, XIAO
Chinese Journal of Experimental Ophthalmology 2013;(1):70-74
Background Diabetic complication is associated with lipid peroxidation.Aldehyde dehydrogenases (ALDH) catalyze the irreversible oxidation of a variety of biological aldehydes,including lipid-derived aldehydes (LDAs),and thus protect organs and tissues from toxic LDAs.Understanding the activity of ALDH in different ocular tissues in diabetic subjects is very important for prevention and treatment of diabetic ocular complications.Objective This research aimed to investigate the activity and expression of ALDH in different ocular tissues in diabetic rats and to explore the mechanism of ALDH in diabetes-induced eye disease.Methods Twenty-eight healthy SPF male Sprague-Dawley(SD) rats weighted 170-180 g were randomly divided into the normal control group and diabetic group.The diabetic animal model was established by intraperitonial injection of 4% streptozotocin at 65 mg/kg.Isometric citric acid buffer was injected in the rats of the normal control group.The rats were sacrificed in each group 2 and 4 months after the establishment of the diabetic models,and eyeballs were obtained for the preparation of corneal,lens and retinal homogenates.ALDH activity was detected using a multifunctional microplate reader SpectraMax M5,and ALDH content was measured by ELISA at the wavelength of 450 nm with the SpectraMax M5 ELISA reader.Results The blood glucose level in diabetic rats was significantly elevated at various time points compared with the normal control group(P=0.000),and body weights were evidently lower in the diabetic group than in the normal group (P =0.000).The activities of ALDH (A340) in corneal,lens and retinal tissues in the diabetic group were increased in comparison with the normal control group (F =396.601,P=0.000),and showed an enhancement with the lapsing of time (F =53.139,P =0.000).In addition,the highest level of ALDH was found in the cornea and the lowest level in the lens(F =6973.000,P=0.000).The expression level of ALDH in the corneal,lens and retinal homogenates was significantly higher in the diabetic group compared with the normal control group (F=312.985,P =0.000) and showed a considerable increase over the course (F =19.203,P=0.000).The highest expression level was seen in the cornea and the lowest was in the lens,with a significant difference among these three kinds of tissues (F =3243.000,P =0.000).Conclusions ALDH can protect ocular tissue from the damage of lipid peroxidation.Thess results suggest that ALDH plays a role in preventing diabetes-related ocular complications.
5.Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts.
Ke, TAO ; Hui, ZENG ; De-ming, XIAO ; Ao, XIONG ; Jian, WENG ; Bin, KANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):241-6
Effect of interleukin-6 receptor (IL-6R) antibody on polymethyl methacrylate (PMMA) bone cement-mediated expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in synovial fibroblasts was investigated. Synovial tissue obtained from total knee arthroplasty was digested and cultured. Inverted microscope was employed to observe the synovial cells and immunocytochemistry (SABC method) staining was used to identify synovial fibroblasts. This experiment was divided into three groups according to different culture media: PMMA group (75 μg/mL PMMA bone cement particles), IL-6R antibody group (10 ng/mL IL-6R antibody+75 μg/mL PMMA bone cement particles), and control group (no IL-6R antibody or PMMA bone cement particles). Influence of IL-6R antibody and PMMA on proliferation of synovial fibroblasts was measured by cell counting kit-8 (CCK-8). ELISA method was used to measure OPG and RANKL levels in culture solution. Fluorescence quantitative real-time PCR (FQ-PCR) was used to detect the expression of OPG and RANKL mRNA. After three consecutive passages, more than 95% of the primary synovial cells became long spindle fibroblast-like cells. SABC staining results showed that the fibroblast-like cells were negative for anti-CD68 antibody and positive for anti-vimentin antibody, with brown madder stained. CCK-8 test demonstrated that the absorbance (A) value at 450 nm was significantly lower in IL-6R antibody group than in PMMA group and control group (P<0.01), but there was no statistically significant difference in A value at 450 nm between the control group and PMMA group (P>0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group (P<0.01). The expression of RANKL was inhibited (P<0.05), and the ratio of OPG/RANKL was significantly increased in IL-6R antibody group as compared with PMMA group and control group. There was no significant difference in the expression of OPG between control group and PMMA group (P>0.05), but the expression of RANKL was higher in PMMA group than in control group (P<0.05), and there was a significant difference in the ratio of OPG/RANKL between them (P<0.05). Results of FQ-PCR revealed the expression of RANKL mRNA was significantly inhibited (P<0.01) and the expression of OPG mRNA was significantly increased (P<0.01) in IL-6R antibody group as compared with PMMA group and control group. The expression of RANKL mRNA was higher in PMMA group than in control group (P<0.05), but the expression of OPG mRNA had no significant difference between them (P>0.05). IL-6R antibody could significantly increase the expression of OPG, but inhibit the expression of RANKL, which might provide a theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of prosthesis.
6.Changes of urine enzyme series and analysis of the related factors in 4 clinical types of purpuric nephritis
xiao-yan, LEI ; hai-jin, XIONG ; hong, ZHANG ; yi-pa, SAI ; jian-jun, WANG
Journal of Applied Clinical Pediatrics 1986;0(01):-
control group from the high to the low. When the typeⅢand type Ⅳglomerular function was changed (BUN and BCr in high value),the drainage quantity of the enzymes evidently increased.Conclusions Urine enzyme series can sensitively reflect the damage of renal tubules in early stage, even if BUN and BCr value is on the normal level , and the drainage quantity of these enzymes are changed more or less, which show that renal tubule damage exists. The value of BUN and BCr is positively correlated with the drainage quantity of these enzymes.the more urine enzymes are drained out, the more renal tubule function is involved, therefore, the more renal globe function is damaged.
7.Immunigical effect of CpG oligodeoxynucleotide as immune adjuvant of hepatitis B vaccine on pregnant mice and neonatal mice
xin, XIAO ; chun-guang, XU ; ai-hua, XIONG ; jian-wei, JIANG ; yan-fang, XU
Journal of Applied Clinical Pediatrics 1992;0(06):-
0.05); serum HBsAb levels of pregnant mice and neonatal mice in group with CpG-1826 (20 ?g)+hepatitis B vaccine significantly higher than those in group with CpG-1826 (10 ?g, 40 ?g)+ hepatitis B vaccine,hepatitis B vaccine and control respectively(P0.05).Conclusions Combination injection of CpG-1826 20 ?g and hepatitis B vaccine can markedly increase serum antibody levels of pregnant mice and neonatal mice, but don′t affect the survival quantity, the growth and development of neonatal mice.CpG-1826 is an ideal immune adjuvant for neonates with immature immune system during pregnancy.
8.Cartilage-ectodermal dysplasia in a case.
Xiao-lu MA ; Bin-xiong XIA ; Ji-yan ZHENG ; Jian SHEN
Chinese Journal of Pediatrics 2004;42(10):799-799
10.Inhibitory effect of polydatin on expression of toll-like receptor 4 in ischemia-reperfusion injured NRK-52E cells.
Ying LI ; Wei-Jian XIONG ; Jing YANG ; Jin ZHONG ; Jin ZHENG ; Ling ZHANG ; Xiao-Qin OUYANG
China Journal of Chinese Materia Medica 2014;39(16):3157-3161
Polydatin is a monocrystaline compound isolated from Polygonum cuspidatum Sieb. et Zucc. (Polygonaceae) with biological properties, such as anti-inflammation, anti-oxidative and nephroprotective effects. Increasing number of studies have demonstrated the protective effect of polydatin on renal ischemia reperfusion injury. However, the possible mechanisms of this protection are not fully elucidated. This study aimed to investigate the effect of polydatin on ischemia-reperfusion induced expression of toll-like receptor4 (TLR4) in rat renal tubular epithelia cells (NRK-52E), and analyze the mechanism of polydatin on TLR4 signal pathway. The cultured NRK-52E cells were incubated in three gas incubators for a period of 6 h at hypoxia and 24h at reoxygenation to simulate the ischemia-reperfusion injury in vitro. TLR4 mRNA level was analyzed by real-time-PCR, and the protein expression of TLR4 and NF-κB by Western blotting, while TNF-α and IL-1β proteins expressions were detected by ELISA. Polydatin downregulated I/R induced mRNA and protein expressions of TLR4, and decreased the protein expression of NF-κB, TNF-α and IL-1β. The TLR4 blocker partially antagonized the effect of I/R on NF-κB signaling, and such inhibitory effect was markedly enhanced by polydatin. In the present study, polydatin protects NRK-52E cells from I/R injury possibly by relieving the inflammatory response through regulation of TLR4/NF-κB signaling pathway.
Animals
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Cell Line
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Drugs, Chinese Herbal
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pharmacology
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Gene Expression
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drug effects
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Glucosides
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pharmacology
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Humans
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Rats
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Reperfusion Injury
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drug therapy
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genetics
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metabolism
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Stilbenes
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pharmacology
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Toll-Like Receptor 4
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genetics
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metabolism