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·AIM: To investigate the role of small interference RNA interference targeted Integrin-linked kinase (ILK SiRNA) on the proliferation and apoptosis of human Tenon fibroblasts (HTFs) induced by transforming growth factor-β2(TGF-β2). · METHODS: The HTFs were identified by immunofluorescence analysis with Vimentin and keratin. HTFs with no other addiction was as normal control;H+T group:HTFs+5μ g/L TGF-β2;H+T+NC SiRNA group:HTFs+5μ g/L TGF-β2+50nmol/L negtive SiRNA; H+T+ILK SiRNA group:HTFs+5μ g/L TGF-β2+50nmol/L ILK SiRNA. The ILK SiRNA were transfected into HTFs by lipofectamine 2000, then the cells were stimulated with 5μ g/L TGF - β2. The protein expression of ILK were analyzed by Western Blot. The proliferation levels of HTFs were analyzed by CCK-8, the apoptosis of HTFs were analyzed by Hoechst 33342/PI double staining. ·RESULTS: The protein ILK were expressed in both TGF-β2treated and control groups, and TGF- β2up-regulated the expression of ILK, ILK SiRNA inhibited the protein expression of ILK(P< 0. 05). CCK- 8 analysis showed that compared with the normal control group,the cell proliferation rate of HTFs in TGF-β2treated group increased, and in ILK SiRNA group the cell proliferation rate was suppressed after exposured to ILK SiRNA for 48h (P<0.05). Hoechst 33342/PI double staining showed that there was no change on the apoptosis of TGF - β2 stimulated group (P>0.05), compared with the normal control group, however in the ILK SiRNA group, we found lots of apoptosis cells and a few of necrotic cells (P<0.05). ·CONCLUSION: The ILK SiRNA attenuates the abnormal proliferation of HTFs induced by TGF - β2, thereby enhancing the apoptosis of HTFs.
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Abstract?AlM: To investigate the effect of zebularine ( Zeb ) loaded Poly ( ethylene glycol ) - block - poly ( ε -caprolactone) methyl ether ( MePEG-PCL) nanoparticles ( NPs) on the viability, attachment, and apoptosis of in vitro cultured lens epithelial cells ( LECs) .?METHODS: In vitro cultured infant human lens tissue HLE B-3 immortalized cells were distributed randomly divided into six groups. Each group was administered with free Zeb 50μmol/L ( ZebF1 group ) , 100μmol/L ( ZebF2 group) , Zeb -loaded MePEG-PCL NPs 50μmol/L ( ZebNP1 group) , Zeb -loaded MePEG-PCL NPs 100μmol/L ( ZebNP2 group) , MePEG-PCL empty NPs( NPs group) or blank medium (group C) respectively. A tetrazolium dye assay ( MTT) test and modified MTT test were performed to determine cell viability and cell attachment. DNA ladder was used to detect the cell apoptosis.?RESULTS: Determined by MTT colorimetric method:Cell proliferation rate of LECs were suppressed by all Zeb administration groups in a concentration-time dependent manner (P ZebNP1>ZebF2 (P<0. 05).?CONCLUSlON: Zeb loaded MePEG-PCL NPs had better effect on suppressing the viability and attachment of in vitro cultured LECs than the free Zeb groups , as well as enhancing the apoptosis.
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AlM: To investigate expressions of matrix metalloproteinase-2 ( MMP-2 ) and extracellular matrix metalloproteinase inducer ( EMMPRlN) in retinoblastoma (Rb) and the relationships between MMP-2, EMMPRlN and tumor development.METHODS:lmmunohistochemical technique was used to detect expressions of MMP-2 and EMMPRlN in 39 cases of paraffin embedded Rb samples. Quantitative analysis of expressions of MMP-2 and EMMPRlN were assessed by measuring the mean gray scale of Rb tissue with LElCA lM50 Color Pathologic Analysis System. The differences of expressions of MMP-2 and EMMPRlN in each clinical and pathological stage were statistically analyzed, and the same step was also undertaken to study the relationship between Rb with MMP-2 positive expression and that with EMMPRlN positive expression.RESULTS:The positive expression rate of MMP-2 was 90% (Gray value: 109. 64 ± 14. 52; 35/39), and that of EMMPRlN was 85% (Gray value:108. 01±13. 60;33/39). The expressions of MMP - 2 and EMMPRlN were significantly higher in tumors of glaucomatous stage (Gray value:108. 21±11. 47 and 107. 56±14. 32) than those in intraocular stage ( Gray value: 121. 13 ± 11. 32 and 119. 34 ± 12. 66; P<0. 01 and P<0. 05). And the same conclusion can be concluded between those in extraocular stage (Gray value: 91. 03 ± 11. 71 and 92. 26 ± 12. 93) with those in glaucomatous stage (P<0. 01 and P<0. 05). The expressions of MMP-2 and EMMPRlN were significantly higher in tumors with optic nerve invasion (Gray value:103. 89±13. 39 and 105. 23±14. 00) than those without optic nerve invasion ( Gray value: 118. 39 ± 15. 11 and 117. 53±16. 13) (P<0. 01 and P<0. 05).CONCLUSlON:The positive expression levels of MMP-2 and EMMPRlN may correlate with tumor infiltration and metastasis.
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AlM:To observe the effects of double incision combined surgery of single - stab trabeculectomy and phacoemulsification. METHODS: Totally 28 cases ( 30 eyes ) with glaucoma and cataract undertook the modified combined surgery of single - stab trabeculectomy and phacoemulsification. After traditional phacoemulsification, cut the bulbar conjunctiva and Tenons capsule from the 11 o'clock to 1 o'clock, then puncture into the anterior chamber in 2mm behind the corneal limbus with 3mm tunnel knife, shaping a 3mm wide, 1/3-1/4 thickness scleral tunnel. Getting into the trabecular tunnel, bite off 3 pieces of trabecular tissue about 1mm × 1mm size. The changes in the imtraocular pressure ( lOP ) and the visual acuity before and after the surgery as well as filtering bleb ( OCT confirmed) and complications were carefully observed in 3-6mo postoperatively. RESULTS: The postoperative visual acuity in 1wk postoperatively less than 0. 1 was found in 3 eyes, from 0. 1 to 0. 3 was found in 6 eyes,from 0. 3 to 0. 6 in 13 eyes, from 0. 6 to 0. 8 in 8 eyes. One eye had malignant glaucoma, and 8 eyes had cornea edema and slightly fibrin exudation in the pupil area; ln all cases maintained function conjunctival blebs of filtering, OCT confirmed this. lOP remained normal in 28 eyes in 3-6mo follow up, lOP of 2 other eyes could be controlled by anti-glaucoma eye drops. CONCLUSlON:Double incision combined surgery of single- stab trabeculectomy and phacoemulsification is effective and safe, reduces the postoperative complications and is worthy of promotion.