Yong-Xiang Jiang

Fudan University, Shanghai, Shanghai Shi, China

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Publications (10)11.43 Total impact

  • Yong-Xiang Jiang, Yi Lu, Tian-Jin Liu, Jin Yang, Xin-Hua Wu
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    ABSTRACT: OBJECTIVE: To explore the specific expression of HSV-tk gene and killing effects on ocular leading cells of the enhanced specific HSV-tk/GCV gene therapy system regulated by lens-specific promoter LEP503. METHODS: Experimental research. The enhanced specific HSV-tk/GCV gene system of two vectors were constructed (Lenti-LEP503-HSVtk-Cre and Lenti-HPGK-Loxp-EGFP-pA-Loxp-HSVtk). The lentiviral vectors were produced by transient transduction of transfering vectors, packaging vectors and enveloping vector into 293T cells. Virus was collected with ultracentrifugation and resuspended with 1 ml phosphate buffered saline and stored at -80°C. The HLEC and RPEC, NIH3T3, 293T cells were transduced with the enhanced specific HSV-tk gene system. The specific expressions of EGFP and HSV-tk were detected by fluorescence microscopy, flow cytometry and RT-PCR. The killing effects of HLEC and RPEC at the concentration of 20 mg/L GCV were assayed and compared by flow cytometry and CCK-8 kit. Difference of RPE cell viability among groups was evaluated by analysis of variance (ANOVA). RESULTS: Expression efficiency of EGFP in RPEC group was 62.3%, 68.3% in NIH3T3 group, 75.8% in 293T group, whereas 17.5% in HLEC group. There was higher expression of HSV-tk at mRNA level in HLEC group than that in RPEC group. The relative intensity of HSV-tk mRNA in HLEC group transduced with the enhanced specific HSV-tk gene system was 4.01, whereas 0.29 in RPEC group. At the concentration of 20 mg/L GCV after 72 hours, the percentage of apoptosis detected by the flow cytometry in HLEC group transduced by the enhanced specific HSV-tk gene system was 76.51%, and 2.44% in RPEC group. There was no significant difference in the RPE cell viability among the enhanced specific HSV-tk gene combination-RPE group, normal-RPE group and negative-RPE control group at the concentration of 20 mg/L GCV after 72 hours (MD(1) = -0.047, P = 0.671; MD(2) = 0.027, P = 0.912). CONCLUSIONS: The enhanced specific HSV-tk gene system express HSV-tk selectively in HLEC. At the concentration of 20 mg/L GCV, it is effective against the proliferation of HLEC in vitro, but has less kill effect on RPEC.
    [Zhonghua yan ke za zhi] Chinese journal of ophthalmology 09/2012; 48(9):829-835.
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    Xin-Hua Wu, Yi Lu, Yan-Wen Fang, Yong-Xiang Jiang
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    ABSTRACT: To investigate whether apoptosis of human lens epithelial cells (HLECs) can be induced with the polyamidoamine (PAMAM)-mediated inhibition of bcl-2 (b-cell lymphoma 2) by small hairpin RNA (shRNA). HLECs (SRA01/04) were transfected with the fifth generation of PAMAM (PAMAM G5) by bcl-2 shRNA. At 24, 48, and 72 h after transfection, the transfection rate was measured by flow cytometry. The transfection rates mediated by PAMAM and liposome were compared. The bcl-2 mRNA level was detected by real-time PCR. Whole cell protein was extracted and the bcl-2 protein level was detected by western blotting. The percentage of HLECs undergoing apoptosis was measured by Annexin V-FITC/PI staining. The nuclear morphology of HLECs was observed by staining with Hoechst 33258. The expression of cytochrome c and the activity of cleaved caspase-3 were analyzed by western blotting. At 24, 48, and 72 h after transfection, the rate of transfection of bcl-2 shRNA mediated by PAMAM was higher than in the liposome-mediated group (p<0.05). The mRNA and protein levels of bcl-2 were greatly downregulated. The percentage of HLECs undergoing apoptosis was greatly improved. Hoechst staining showed that bcl-2 shRNA transfected cells had a lower growth status with nuclear fragmentation. The expression of cytochrome c and the activity of cleaved caspase-3 was greatly improved (p<0.05). PAMAM-mediated bcl-2 shRNA can downregulate the expression of bcl-2 and induce the apoptosis of HLECs by engaging the mitochondrial pathway, including catalytic activation of the caspases.
    Molecular vision 01/2012; 18:74-80. · 1.99 Impact Factor
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    ABSTRACT: Cataract and geographic atrophy (GA, also called advanced "dry" age-related macular degeneration) are the two major causes of visual impairment in the developed world. The association between cataract surgery and the development of GA was controversial in previous studies. We performed a meta-analysis by pooling the current evidence in literature and found that cataract is associated with an increased risk of geographic atrophy with a summary odds ratio (OR) of 3.75 (95% CI: 95% CI: 1.84-7.62). However, cataract surgery is not associated with the risk of geographic atrophy (polled OR=3.23, 95% CI: 0.63-16.47). Further experiments were performed to analyze how the αA-crystallin, the major component of the lens, influences the development of GA in a mouse model. We found that theαA-crystallin mRNA and protein expression increased after oxidative stress induced by NaIO(3) in immunohistochemistry of retinal section and western blot of posterior eyecups. Both functional and histopathological evidence confirmed that GA is more severe in αA-crystallin knockout mice compared to wild-type mice. Therefore, αA-crystallin may protect against geographic atrophy. This study provides a better understanding of the relationship between cataract, cataract surgery, and GA.
    PLoS ONE 01/2012; 7(8):e43173. · 3.53 Impact Factor
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    Jin Yang, Tian-Jin Liu, Yong-Xiang Jiang, Yi Lu
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    ABSTRACT: To establish a novel, targeted lentivirus-mediated LEP503-HSV-tk/GCV suicide gene therapy system combined with all trans-retinoic acid (ATRA) for the inhibition of human lens epithelial cell (HLEC) proliferation and treatment of posterior capsular opacification (PCO) after cataract surgery; to estimate the enhancement of the bystander effect by ATRA; and to explore the role of Connexin43 (Cx43) mediated gap junctional intercellular communication (GJIC) in the bystander effect of the HSV-K/GCV system. A Lenti-LEP503-HSV-tk-EGFP vector was generated by cloning the lens-specific promoter LEP503 (lens specific promoter 503) from genomic DNA of HLECs by PCR. The vector was then inserted into the promoter-less vector from lentivirus-based (CMV)-HSV-tk-EGFP. The expressional specificity of the LEP503 promoter was assessed by investigating the expression of EGFP (enhanced green fluorescent protein) and HSV-tk (herpes simplex virus thymidine kinase) mRNA, both driven by Lenti-LEP503-HSV-tk-EGFP vector, by fluorescence microscopy, RT-PCR, flow cytometry, and western blot assays in HLECs, human adult retinal pigment epithelium cells (RPECs), human adult skin fibroblast cells (ASFCs), and Hela cells. Morphological changes were observed by fluorescence microscopy and cell viability was determined using the Cell Counting kit-8 Cell Proliferation (CCK-8) and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays after Lenti-LEP503-HSV-tk/GCV system combined with ATRA treatment on HLECs. Flow cytometry, DNA fragmentation, and western blot assays were employed to analyze the mechanisms of bystander effects. The promoter LEP503-mediated HSV-tk was specifically expressed in HLECs, and ATRA dose-dependently strengthened the bystander effect following LEP503-mediated HSV-tk/GCV gene therapy against lens cells by upregulating the expression of the gap junction protein Cx43. The Lenti-LEP503-HSV-tk/GCV suicide gene therapy system, combined with ATRA as an adjuvant, may be a feasible supplementary method for PCO treatment that targets residual lens cells.
    Molecular vision 01/2012; 18:2053-66. · 1.99 Impact Factor
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    ABSTRACT: To establish a novel, targeted lentivirus-based HSV-tk (herpes simplex virus thymidine kinase)/GCV (ganciclovir) gene therapy system to inhibit lens epithelial cell proliferation for treatment of posterior capsular opacification (PCO) after cataract surgery. An enhanced Cre recombinase (Cre/loxP) system with a lentiviral vector expressing Cre under the control of the lens-specific promoter LEP503 (Lenti-LEP503-HSVtk-Cre [LTKCRE]) was constructed, as well as another lentiviral vector containing a switching unit. The latter vector contains a stuffer sequence encoding EGFP (Lenti-hPGK-Loxp-EGFP-pA-Loxp-HSVtk [PGFPTK]) with a functional polyadenylation signal between two loxP sites, followed by the herpes simplex virus thymidine kinase (HSV-tk) gene, both under the control of the human phosphoglycerate kinase (hPGK) promoter. Expression of the downstream gene (HSV-tk) is activated by co-expression of Cre. Human lens epithelial cells (HLECs) or retinal pigmental epithelial cells (RPECs) were co-infected with LTKCRE and PGFPTK. The inhibitory effects on HLECs and RPECs infected by the enhanced specific lentiviral vector combination at the concentration of 20 µg/ml GCV were assayed and compared. The specific gene expression of Cre and HSV-tk in HLECs is activated by the LEP503 promoter. LTKCRE and PGFPTK co-infected HLECs, but not RPECs, expressed high levels of the HSV-tk protein. After 96 h of GCV treatment, the percentage of apoptotic HLECs infected by the enhanced specific lentiviral vector combination was 87.23%, whereas that of apoptotic RPECs was only 10.12%. Electron microscopy showed that GCV induced apoptosis and necrosis of the infected HLECs. The enhanced specific lentiviral vector combination selectively and effectively expressed HSV-tk in HLECs. A concentration of 20 µg/ml, GCV is effective against the proliferation of HLECs in vitro. This cell-type-specific gene therapy using a Cre/loxP lentivirus system may be a feasible treatment strategy to prevent PCO.
    Molecular vision 01/2011; 17:291-9. · 1.99 Impact Factor
  • Ying-hong Ji, Yi Lu, Lin Wang, Yong-xiang Jiang
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    ABSTRACT: To evaluate the visual performance after implantation of the Tecnis ZM900 multifocal intraocular lens (IOL) (TMF) and the Restor SA60D3 multifocal IOL (Restor). In a prospective study, TMF or Restor was implanted randomly in 73 patients (90 eyes). The following parameters were assessed 3 months after surgery: refraction, uncorrected and best corrected visual acuities (VA) for distance, intermediate, near and different contrast levels, reading ability, pupil size, wave-front error, defocus curve and position of IOL. Patient satisfaction (overall satisfaction, spectacle independence, photic phenomena) was assessed by a questionnaire. The chi-square test was applied to compare categorical variables and Mann-Whitney U test was used to compare the measured data. The uncorrected, best corrected and/or distance-corrected VA for distance, intermediate, near and different contrast levels did not show statistically significant differences between the two groups (P > 0.05). Near reading acuity and reading speed were better in TMF under low-light conditions (Z = -2.579, P = 0.009; Z = -5.244, P = 0.000). The curve of defocus showed that TMF had significantly better intermediate distance (at 50 cm) (Z = -5.300, P = 0.000) and worse near distance (from 25 to 28 cm) than those of Restor (Z = -3.745, P = 0.000; Z = -5.691, P = 0.000). Measurements under pupil diameter at 3.0 mm and 5.0 mm, ocular and intraocular Z (4, 0) were significantly lower (Z = -8.175, P = 0.000; Z = -5.210, P = 0.000 and Z = -4.453, P = 0.000; Z = -3.790, P = 0.000), the values of PSF Strehl Ratio and MTF AreaRatio A/D were significantly higher (Z = -3.047, P = 0.002; Z = -3.672, P = 0.008 and Z = -2.038, P = 0.042; Z = -2.579, P = 0.009) in TMF than those in Restor. On the questionnaire, there was no difference of overall satisfaction, spectacle independence and photic phenomena (P > 0.05). Implantation of the TMF and Restor offers excellent distant and near VA. Restor had better near VA than that of TMF based on the curve of depth, TMF had better VA at 50 cm-distance. Reading speed is faster in TMF. Compared to spherical Restor, TMF provides a better quality of vision due to a negative spherical aberration.
    [Zhonghua yan ke za zhi] Chinese journal of ophthalmology 08/2010; 46(8):679-85.
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    ABSTRACT: To evaluate the results of cataract surgery in myopia patients after laser in situ keratomileusis (LASIK) and to compare the predictability of various methods of intraocular lens (IOL) power calculation. Seventeen cases (24 eyes) who had LASIK for myopia were divided into two group by with or without history of corneal power data. Corneal power was obtained by autokeratometry, corneal topography, Pentacam and IOL Master. The IOL power was calculated with the clinical history method, Feiz-Mannis formula, Feiz-Mannis method and other methods. Postoperative final refraction and the deviation of the final spherical equivalent (SEQ) from the refractive target were measured 3 month after the surgery. Two sample t-test, linear correlation and regression analysis, paired t-test and Bland-Altman method of agreement were used to analyze these data. In the group with history data, the mean corneal power was (43.28 ± 1.21) D and the mean SEQ was (-15.33 ± 4.36) D before the LASIK surgery. In the group without history data, the mean SEQ was (-10.11 ± 3.12) D. Before cataract surgery, the mean corneal power was (36.96 ± 2.07) D and (36.85 ± 1.40) D in these two groups. The mean arithmetic refractive prediction error after cataract surgery was (-0.66 ± 1.27) D and (-0.47 ± 0.82) D in these two groups, respectively. Data calculated by using Hamed-Wang-Koch method, Masket Formula, Koch/Maloney method, Shammar method and Pentacam ERK method were lower than the emmetropic IOL power. Data calculated by using Feiz-Mannis Formula, Latkany Method, Savini method, Armberri Double K method were overestimated. The mean arithmetic errors of clinic history method, Corneal Passby Method and Haigis-L Formula were not significantly different from the predict refraction (P = 0.364, 0.318 and 0.069; t = 0.956, -1.057 and -1.911, respectively). There was strong correlation between the value calculated by using Feiz-Mannis Method or Haigis-L Formula and the true power (r = 0.921, 0.915; P = 0.000 and 0.000, respectively). But none of the values calculated by these method could fully agree with the true value. IOL power should be calculated accurately to avoid undercorrection. We recommend the combination of clinical history method, Feiz-Mannis Method, Corneal Passby Method and Haigis-L Formula for the calculation of IOL power.
    [Zhonghua yan ke za zhi] Chinese journal of ophthalmology 06/2010; 46(6):518-24.
  • Yi Lu, Ying-Hong Ji, Yi Luo, Yong-Xiang Jiang, Man Wang, Xu Chen
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    ABSTRACT: To present the visual results and the complications of primary intraocular lens (IOL) implantation in infants aged 6 to 12 months between January 2002 and July 2007. A total of 26 consecutive eyes, of 16 infants with cataract aged 6 to 12 months, were reviewed in the study. All patients had cataract extraction with anterior and posterior capsulorrhexis combined with anterior vitrectomy and primary hydrophobic acrylic IOL implantation. Six infants (six eyes) had unilateral congenital cataract and ten (20 eyes), bilateral cataract. Visual acuity and complications were recorded throughout the 46.4-month mean follow-up (range 22 to 79 months). All eyes had primary IOL implantation. The mean best-corrected visual acuity (logMAR) was 0.98 +/- 0.18,0.50 +/- 0.14 and 0.61 +/- 0.25 for unilateral, bilateral and all eyes respectively at the last follow-up. IOLs were implanted in the capsular bag of 25 eyes (96.2%) and in the sulcus of the remaining one eye (3.8%). Seven eyes (26.9%) developed visual axis opacification (VAO), and four eyes required secondary pars plana vitrectomy (PPV). IOL opacification occurred in one eye 54 months after implantation. Late onset open-angle glaucoma developed in one eye, and required trabeculectomy surgery. The predictors of good best-corrected visual acuity (BCVA) included partial cataract, bilateral cataract, absence of strabismus or nystagmus, and good amblyopic treatment. The greatest annual myopic change (5.15 +/- 2.08 D) was observed during the first 12 months after surgery. In unilateral cases, there was no significant difference in the axial length between the cataractous eye and the fellow normal eye both at the time of surgery (P = 0.891) and final follow-up (P = 0.693). Primary IOL implantation was safe and effective for infantile cataract surgery. Total or unilateral cataract, nystagmus or strabismus, and inadequate amblyopic therapy were predictors of poor BCVA. Significant myopic shifts occurred especially in infants in the first year of surgery. The pseudophakic eye had a similar growth rate, as measured by axial length, to that of the fellow normal eye, in unilateral cases.
    Albrecht von Graæes Archiv für Ophthalmologie 02/2010; 248(5):681-6. · 1.93 Impact Factor
  • Xin-Hua Wu, Yi Lu, Yan-Wen Fang, Yong-Xiang Jiang, Jie Tian
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    ABSTRACT: To investigate whether apoptosis of human lens epithelial cells (HLEC) can be induced by inhibition of bcl-2 shRNA. It was an experimental study. Two pairs of oligonucleotides were synthesized and inserted into plasmid PGCsi to generate shRNA eukaryotic expression vectors, named P1 and P2. At 48 hours after transfection in HLEC (SRA01/04) with Lipofectamine 2000, the whole cell protein was extracted and detected by Western blot. The bcl-2 mRNA level was detected by real-time PCR. The percentage of HLECs undergoing apoptosis was measured by Annexin V-PI staining. The activity of caspase-3 was analyzed by Western blotting. The shRNA eukaryotic expression vectors were constructed successfully. At 48 hours after transfection, the rate of transfection of P1 and P2 was about 44.1% and 47.2% respectively. The protein and mRNA level of bcl-2 was 0.435 and 0.476, greatly downregulated (F = 1672.4, P < 0.05). The percentage of HLEC undergoing apoptosis was 42.3% and 45.4%, greatly improved (F = 1756.2, P < 0.05). The activity of caspase-3 was greatly improved. P1 and P2 can both down-regulate the expression of bcl-2, and induce the apoptosis of HLEC.
    [Zhonghua yan ke za zhi] Chinese journal of ophthalmology 07/2009; 45(7):636-40.
  • Zhi-gang Lü, Wen-li Huang, Yong-xiang Jiang, Tian-jin Liu
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    ABSTRACT: To explore the expression of telomerase activity of the lens epithelial cells in posterior capsule opacification of rabbits. Clear corneal tunnel phacoemulsification was performed in both eyes of 11 New Zealand rabbits. After the model of posterior capsule opacification succeed in all eyes. Then, telomerase activity of the lens epithelial cells in the equator and posterior capsule opacification of 20 eyes was detected with TRAP-ELISA and TRAP-PAGE techniques. HepG2 cells were used as positive controls. Telomerase activity was detected in the lens epithelial cells in the equator capsule and posterior capsule opacification in rabbits, which showed several dim gradient stripped electrophoresis pattern. A450 - 690 value of telomerase activity in the equator capsule and posterior capsule opacification was 0.85 +/- 0.23 and 0.67 +/- 0.19, respectively, indicating statistically significant difference (t = 2.526, 0.021; P < 0.05). Telomerase activity exists in the lens epithelial cells of posterior capsule opacification in rabbits. The telomerase activity in this area is lower than that in the equator capsule.
    [Zhonghua yan ke za zhi] Chinese journal of ophthalmology 11/2008; 44(10):902-5.