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Gao X.-J.,Peoples Hospital of Sichuan Province | Gao X.-J.,Chongqing Key Laboratory of Ophthalmology | Hou S.-P.,Chongqing Medical University | Hou S.-P.,Chongqing Key Laboratory of Ophthalmology | And 2 more authors.
International Journal of Ophthalmology | Year: 2014

AIM: To examine the association between the single nucleotide polymorphisms (SNPs) of matrix metalloprotease-9 (MMP-9) gene and primary angle-closure glaucoma (PACG) in a Chinese Han population. METHODS: DNA samples were extracted from peripheral-blood mononuclear cells of 214 PACG patients and 224 healthy controls. Genotyping of rs3918249, rs3918254, rs17577 and rs3787268 in MMP-9 was performed using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and the direct sequencing technique. The association between these genetic polymorphisms and risk of PACG was estimated by χ2 test. RESULTS: The distributions of rs3918249, rs3918254, rs17577 and rs3787268 genotypes among cases and healthy controls were compatible with that from Hardy-Weinberg equilibrium (HWE, P>0.05). The increased frequency of CC and CT genotypes of rs3918254 were observed in PACG patients compared to healthy controls [P=0.006, Pcorrected (Pcorr)=0.048]. The haplotype analysis showed that the CCGG haplotype was nominal associated wth PACG (P=0.015), however, the significant was lost when the Bonferroni correction was used (Pcorr=0.105). CONCLUSION: Our results revealed that rs3918254 in MMP-9 may be a susceptible locus to PACG in China, people wth the CC and CT genotypes of rs3918254 are more susceptible to PACG. The susceptibility to PACG in Chinese Han patients may be not influenced by SNPs rs3918249, rs3787268 and rs17577 in MMP-9. Copyright International Journal of Ophthalmology Press.


Yang Y.,Chongqing Medical University | Yang Y.,Chongqing Key Laboratory of Ophthalmology | Yang Y.,Chongqing Eye Institute | Yang Y.,Sun Yat Sen University | And 14 more authors.
Investigative Ophthalmology and Visual Science | Year: 2012

PURPOSE. IL-7/IL-7R has been found to be involved in the pathogenesis of several autoimmune diseases. This study was designed to investigate the potential role of IL-7/IL-7R in the pathogenesis of Vogt Koyanagi-Harada (VKH), an organ-specific autoimmune disease. METHODS. IL-7 was measured with an enzyme-linked immunosorbent assay (ELISA) in serum obtained from patients with active or inactive VKH and from healthy individuals. The expression of IL-7R was measured by flow cytometry (FCM). Cell proliferation was determined after exposure of peripheral blood mononuclear cells (PBMCs) and CD4 + T cells to recombinant IL-7. The levels of IL-17 and IFN-_ levels were detected by ELISA after these cells were cocultured with recombinant IL-7. The influence of recombinant IL-7 on the expansion of Th1 and Th17 cells was evaluated by using FCM. RESULTS. IL-7 was significantly increased in the serum of patients with active VKH compared with those with inactive VKH (P < 0.001) and normal controls (P < 0.001). However, there was no difference between VKH patients and normal controls in the expression of IL-7R_ on CD4 + T cells. Recombinant IL-7 induced significant cell proliferation and secretion of IL-17 and IFN-γ by PBMCs and CD4 + T cells. It furthermore promoted the expansion of both Th1 and Th17 cells. CONCLUSIONS. The findings suggest that IL-7 is involved in the pathogenesis of VKH disease. © 2012 The Association for Research in Vision and Ophthalmology, Inc.


Hou S.,Chongqing Medical University | Hou S.,Chongqing Key Laboratory of Ophthalmology | Hou S.,Chongqing Eye Institute | Qi J.,Chongqing Medical University | And 15 more authors.
British Journal of Ophthalmology | Year: 2014

Aims Considering the phenotypical consequences and association of C4 copy number variation (CNV) with various autoimmune diseases, we aimed to examine C4 CNVs for 1027 patients with Vogt-Koyanagi-Harada (VKH) syndrome and 2083 controls.Methods C4 CNVs were examined by real-time PCR for 1027 patients with VKH and 2083 controls. Peripheral blood mononuclear cells (PBMC) were prepared from venous blood by Ficoll-Hypaque densitygradient centrifugation for cell culture. Cytokine production was examined by ELISA.Results The expression of total C4 in serum was significantly decreased in patients with VKH as compared with controls (p=0.0010). A significant positive association between C4 expression with C4 CNVs was found (p=0.0023, r2=0.92). CNV analysis identified significantly decreased frequencies of more than two copies of C4A or more than four copies of total C4 in patients with VKH (Pc=1.42×10-3 to 3.56×10 -4, OR=0.67 to 0.70). Linkage analysis showed the independent association of C4 with VKH syndrome from human leucocyte antigen (HLA)-DR4. No significant association was observed concerning type 1 T helper cell (Th1) cytokines and Th17 cytokine production by stimulated PBMCs and C4A copy number.Conclusions Our findings indicate a decreased expression of serum C4 and a decreased frequency of high C4 gene copy number in patients with VKH.Trial registration number Chinese Clinical The authors would like to thank all donors enrolled in the present study.


Hou S.,Chongqing Medical University | Hou S.,Chongqing Eye Institute | Hou S.,Chongqing Key Laboratory of Ophthalmology | Xiao X.,Chongqing Medical University | And 12 more authors.
Human Genetics | Year: 2012

Previous GWAS studies from Turkey suggested a potential risk locus at CCR1/CCR3 for Behçet's disease. However, this locus did not reach the GWAS significance threshold and has not yet been examined in other ethnic populations. The current study aimed to explore whether this locus was associated with Behçet's disease in Chinese Han and the functional role of the identified variants. A two-stage association study was performed in 653 patients and 1,685 controls using the iPLEX system. Real-time PCR was performed to examine the expression level of CCR1 and CCR3 genes. Haplotype analysis was used to construct the haplotype block. Logistic regression analysis was applied to calculate the independence of multiple associations. Bonferroni correction was applied to account for multiple testing. First stage analysis showed that ten SNPs, located in 3'UTR, 5'UTR in CCR1 or 5'UTR in CCR3, were significantly associated with Behçet's disease (Pc = 0.018 to 1.3 9 10-3). The associations of six SNPs within this locus are independent after control for the genetic effect of rs17282391 using logistic regression analysis. Haplotype analysis identified three associated haplotypes: H3 (GTGAC), H6 (CCATTA) and H9 (CGA) (Pc = 0.04 to 7.79 9 10-4). Three SNPs rs13084057, rs13092160 and rs13075270 showed consistent association in replication and combining studies (replication Pc = 5.31 9 10-5 to 1.44 9 10-5; combining Pc = 2.76 9 10-7 to 6.50 9 10-8). Interestingly, eQTLs database reveals that SNP rs13092160 is eQTLs SNP, suggesting that this SNP is likely to be functional SNP that directly affects gene expression. The expression of CCR1 and CCR3 was increased in individuals with the CT genotype of rs13092160 (P<0.05). No significant difference was found for the mRNA level of CCR1 and CCR3 between Behçet's patients and controls. These findings strongly indicate CCR1/CCR3 as a novel locus underlying Behçet's disease. © Springer-Verlag 2012.


Huang X.,Chongqing Medical University | Su Y.-X.,Chongqing Medical University | Deng H.-C.,Chongqing Medical University | Zhang M.-X.,Chongqing Medical University | And 2 more authors.
Hormone and Metabolic Research | Year: 2014

Renal growth, particularly hypertrophy, is a feature of diabetic nephropathy (DN). Adiponectin, an adipocyte-derived hormone, is an important regulator of cell proliferation. Recent studies have suggested that adiponectin has a protective effect in the kidney. The purpose of this study was to investigate the therapeutic effects and the underlying mechanisms of adiponectin in early DN. Mouse mesangial cells (MMCs) were cultured in media containing different concentrations of platelet-derived growth factor-BB (PDGF-BB) with or without adiponectin. MMC proliferation and expression of type IV collagen, laminin, and fibronectin were investigated. Streptozotocin-induced diabetic mice were injected intravenously with recombinant lentivirus encoding the mouse adiponectin gene (Lenti-Acdc-IRES-EGFP). Urinary microalbumin, serum adiponectin level, and expression of proliferating cell nuclear antigen, type IV collagen, laminin, and fibronectin were determined. Adiponectin inhibited the increases in MMC proliferation and expression of type IV collagen, laminin, and fibronectin induced by PDGF-BB. Adiponectin also effectively reduced renal cell proliferation and expression of type IV collagen, laminin, and fibronectin when it was introduced in vivo by lentivirus-mediated gene transfer. These findings suggest that adiponectin exerts renoprotective effects by inhibiting renal cell proliferation and reducing synthesis of extracellular matrix proteins, thus suppressing the development and progression of DN. © Georg Thieme Verlag KG.


Shen Y.,Chongqing Medical University | Yuan X.-D.,Chongqing Medical University | Yuan X.-D.,Chongqing Key Laboratory of Ophthalmology | Hu D.,Chongqing Medical University | And 6 more authors.
Human Immunology | Year: 2014

Objectives: Allergic rhinitis (AR) is an inflammatory disorder of the upper airway. Interleukin-27 (IL-27), a novel IL-12 family member, has recently been reported to play a role in some immune-related disorders. This study was performed to evaluate the potential association of IL-27 polymorphisms with AR in a Chinese Han population. Design and Methods: A case-control study was performed in 445 Chinese AR patients and 691 healthy controls. Three SNPs in the IL-27p28 gene, including rs153109, rs17855750 and rs181206, were detected using a polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP). Results: A significantly increased prevalence of the rs153109 TT genotype and the T allele was found in AR patients, while a decreased prevalence of the CT and CC genotypes and the C allele was found. For rs153109, the TT genotype and the T allele were significantly associated with the risk of AR, but the CT and CC genotypes and the C allele decreased the risk of AR; for rs17855750, the TT genotype and T allele were risk factors for AR, and the GT genotype and G allele provided protection. TTT and TTC haplotypes in the IL-27p28 gene were positively correlated with AR, while CGT, CTC and CTT haplotypes were associated with a significantly decreased risk of AR. Conclusion: This study indicates that IL-27p28 polymorphisms rs153109 and rs17855750 are likely involved in AR susceptibility, making them potentially useful genetic biomarkers for AR susceptibility in the Chinese Han population. © 2014 American Society for Histocompatibility and Immunogenetics.


Zheng M.-M.,Chongqing Medical University | Zheng M.-M.,Chongqing Key Laboratory of Ophthalmology | Zhou X.-Y.,Chongqing Medical University | Zhou X.-Y.,Chongqing Key Laboratory of Ophthalmology | And 3 more authors.
Ultrasound in Medicine and Biology | Year: 2012

The purpose of this study was to explore the transfection of the recombinant expression plasmid pEGFP-C1/RB94 into human retinoblastoma cells (HXO-Rb44) using ultrasound-targeted microbubble destruction (UTMD). pEGFP-C1/RB94 was transfected into HXO-Rb44 in vitro by UTMD, with liposome as the positive control. After 24 to 72 h, the expression of the reporter gene enhanced green fluorescent protein (EGFP) was observed using fluorescent microscopy and flow cytometry. The cell viability of HXO-Rb44 was measured by a MTT assay. The mRNA and proteins of RB94, caspase-3 and Bax were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Moreover, the apoptosis rate and cell cycle progression of the cells were detected by flow cytometry. This study demonstrated that UTMD can enhance the transfection efficiency of RB94, which has an obvious impact on the inhibition of the growth process of retinoblastoma cells, suggesting that the combination of UTMD and RB94 compounds might be a useful tool for use in the gene therapy of retinoblastoma. © 2012 World Federation for Ultrasound in Medicine & Biology.


Tan W.,Chongqing Medical University | Tan W.,Chongqing Key Laboratory of Ophthalmology | Hou S.,Chongqing Medical University | Hou S.,Chongqing Key Laboratory of Ophthalmology | And 8 more authors.
Molecular Vision | Year: 2011

Purpose: The gene for Eph-receptor tyrosinekinase-type A2 (EPHA2) has been shown to be involved in the pathogenesis of age-related cataract (ARC). The aim of this study was to examine whether EPHA2 polymorphisms were associated with the susceptibility to age-related cortical cataract in a Han Chinese population. Methods: Five single-nucleotide polymorphisms (SNPs)-rs3768293, rs3754334, rs7548209, rs707455, and rs477558-in the EPHA2 gene were genotyped in 422 Han Chinese patients with age-related cortical cataract and 317 age-, sex-, and ethnically matched healthy controls using a PCR restriction fragment length polymorphism (PCR-RFLP) assay. Data were analyzed by χ2 analysis. Results: The results showed that the five analyzed polymorphisms in EPHA2 were in Hardy-Weinberg equilibrium both in the patients and in the controls. The frequency of the rs477558 AA genotype was significantly increased in ARC patients compared with controls (χ2=8.649, pc=0.045, odds ratio [OR] 1.555, 95% CI 1.158 to 2.089). The frequency of the rs477558 AG genotype was significantly decreased in ARC patients compared with controls (χ2=9.281, pc=0.030, OR 0.626, 95% CI 0.463 to 0.847). Significantly higher frequencies of the GG genotype and the G allele of rs7548209 were observed in ARC patients compared with controls (χ2=10.430, pc=0.015, OR 1.660, 95% CI 1.219 to 2.261 and χ2=8.537, pc=0.015, OR 1.486, 95% CI 1.138 to 1.940, respectively). On the other hand, significantly decreased frequencies of the rs7548209 CG genotype and the C allele were observed in ARC patients compared with controls (χ2=9.999, pc=0.030, OR 0.603, 95% CI 0.440 to 0.826 and χ2=8.537, pc=0.015, OR 0.673, 95% CI 0.515 to 0.879, respectively). There was no difference in the frequencies of the genotype and allele of the rs3768293, rs3754334, and rs707455 SNPs between the patients with ARC and the controls. Conclusions: Our study suggests that both SNP rs477558 and SNP rs7548209 of EPHA2 are associated with age-related cortical cataract in a Han Chinese population. © 2011 Molecular Vision.


Gao R.,Chongqing Medical University | Gao R.,Dujiangyan Medical Center | Zhou X.,Chongqing Medical University | Yang Y.,Chongqing Medical University | And 2 more authors.
Ultrasound in Medicine and Biology | Year: 2014

Using ultrasound-targeted microbubble destruction (UTMD), we transfected both wild-type p53 (wtp53) and Rb94 genes into retinoblastomas (RBs) of nude mice to investigate the inhibitory role of these two genes in RB development. The 40 tumor-bearing mice, which had been established by sub-retinal injection of an HXO-Rb44cell suspension, were randomly divided into five groups: blank control group (C); blank plasmid group (M); wtp53 plasmid group (p53); Rb94 plasmid group (Rb94); wtp53+Rb94 plasmid group (p53+Rb94). For preparation of the DNA-loaded microbubbles, a pre-determined amount of blank plasmid, pVIVO1-p53, pVIVO1-Rb94 or pVIVO1-p53-Rb94 was added and mixed with the microbubbles. Then, these DNA-loaded microbubbles were respectively transfected into the animal model by UTMD. Vascular endothelial growth factor level and microvessel density of the tumor were determined by immunohistochemical staining. Apoptosis of tissues was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Expression of wtp53 and Rb94 at both the gene and protein levels was detected by RT-PCR (reverse transcription polymerase chain reaction) and Western blot, respectively. Transfection of both genes had greater inhibitory effects on RB development and resulted in lower levels of vascular endothelial growth factor, lower microvessel density and more obvious apoptosis than single-gene transfection (p<0.05). The results indicate that the transfection of both genes into the RB by UTMD more strongly inhibited RB growth than transfection of a single gene. © 2014 World Federation for Ultrasound in Medicine & Biology.


Su Y.-X.,Chongqing Medical University | Deng H.-C.,Chongqing Medical University | Zhang M.-X.,Chongqing Medical University | Long J.,Chongqing Medical University | Peng Z.-G.,Chongqing Key Laboratory of Ophthalmology
Hormone and Metabolic Research | Year: 2012

An aberrant proliferation of mesangial cells (MCs) is one of the more important features of diabetic nephropathy (DN). Adiponectin, an adipocyte-derived hormone, has been associated with type 2 diabetes, a known cause of DN. Recent studies have suggested that adiponectin has a protective effect on the kidney. To elucidate the potential protective mechanism of adiponectin on kidney, we investigated the effects of adiponectin on platelet-derived growth factor (PDGF)-induced cell proliferation and intracellular signaling pathways in cultured Human MCs (HMCs). PDGF-induced HMC proliferation was significantly inhibited by the co-treatment of adiponectin. Adiponectin alone had no effect on HMC proliferation. The mammalian target of rapamycin (mTOR) and 40S ribosomal S6 kinase 1 (S6K1) were activated by PDGF stimulation in HMCs. PDGF-induced mTOR and S6K1 phosphorylations were significantly attenuated by the co-treatment of adiponectin in HMC. Adiponectin alone had no effects on PDGF-receptor autophosphorylation by PDGF. We also confirmed that the inhibitory effect of adiponectin on PDGF-induced HMC proliferation was significantly suppressed by compound C, an adenosine 5-monophosphate-activated protein kinase (AMPK) inhibitor. From these findings, it is implied that adiponectin could attenuate renal dysfunction associated with MC disorders through AMPK-mTOR signal pathway. © Georg Thieme Verlag KG Stuttgart · New York.

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