In order to investigate whether cultured normal human lens epithelial cells (LEC) express transforming growth factor beta (TGF-beta), reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical methods were used for detection of TGF-beta mRNA and protein in cultured normal human LEC. The results showed that a single RT-PCR amplified product about 310bp was obtained, and the sequence was homologous to the known sequence. TGF-beta immunostain was positive in the plasma of LEC. It was suggested that normal human LEC could produce TGF-beta, and LEC could be affected by TGF-beta through autocrine action.
Cells, Cultured ; Epithelial Cells/cytology ; Epithelial Cells/*metabolism ; Lens, Crystalline/cytology ; Lens, Crystalline/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Transforming Growth Factor beta/*biosynthesis ; Transforming Growth Factor beta/genetics

Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kip1-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endothelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combination was used as negative control (pGenesil-HK). The recombination of four plamids was confirmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kip1 was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kip1 mRNA and p27Kip1 protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group). pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The proliferation rates of the pGenesil-P3 group, the pGenesil-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kip1 on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesil-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kip1 could down-regulate the expression of p27Kip1 effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.
Cell Proliferation ; Cornea/cytology ; Cyclin-Dependent Kinase Inhibitor p27/*metabolism ; Endothelial Cells/*cytology ; Endothelial Cells/metabolism ; Gene Expression Regulation ; Models, Biological ; Plasmids/metabolism ; RNA Interference ; RNA, Messenger/metabolism ; RNA, Small Interfering/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tetrazolium Salts/pharmacology ; Thiazoles/pharmacology ; Transfection

The diagnosis and treatment of the lacrimal passage obstruction with lacrimal endoscope was investigated and its subsidiary surgical procedures were evaluated. Ninety-three patients (109 eyes) with lacrimal passage obstruction, including presaccal canalicular obstruction (PSCO) and nasolacrimal duct obstruction (NLDO), were examined under a lacrimal endoscope, and the obstructions were treated with laser or micro-drill. All patients were followed up after the operation for 3-6 months. The difference between the laser and the micro-drill treatment was observed. During the period of follow-up, the curative rate was 82.57%. The healing rate in PSCO group and NLDO was 80.36% and 84.91% respectively (P>0.05). After treatment with the laser, the healing rate was 93.33% in the PSCO group and 66.67% in the NLDO group respectively (P<0.05). After treatment with the micro-drill, the healing rate in PSCO and NLDO groups was 65.39% and 94.28% respectively (P<0.01). The lacrimal passage obstruction can be observed and treated directly through the lacrimal endoscope. Choosing different surgical procedures in operation according to the locations of the obstruction is helpful to improve the effectiveness.
Endoscopy/*methods ; Follow-Up Studies ; Lacrimal Duct Obstruction/*diagnosis ; Lacrimal Duct Obstruction/*surgery ; Laser Therapy/*methods ; Young Adult

In order to investigate whether cultured normal human lens epithelial cells (LEC) express transforming growth factor beta (TGF-beta), reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical methods were used for detection of TGF-beta mRNA and protein in cultured normal human LEC. The results showed that a single RT-PCR amplified product about 310bp was obtained, and the sequence was homologous to the known sequence. TGF-beta immunostain was positive in the plasma of LEC. It was suggested that normal human LEC could produce TGF-beta, and LEC could be affected by TGF-beta through autocrine action.
Cells, Cultured ; Epithelial Cells ; cytology ; metabolism ; Humans ; Lens, Crystalline ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Transforming Growth Factor beta ; biosynthesis ; genetics

Summary: The effects of NO-Fluvastatin on proliferation of human lens epithelial cells (HLECs) and the action mechanism were investigated. Cell proliferation was assessed by MTT assay. Cell cycle was analyzed by flow cytometry. The expression of cell cycle regulatory proteins CyclinE mRNA and P21wafl mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). MTT staining colorimetry showed that HLECs proliferation was markedly inhibited by NO-Fluvastatin and the effect was dependently related to time (24, 48 and 72 h) and dosage (1, 5 and 20 μmol/L). Flow cytometry revealed that NO-Fluvastatin could significantly block HLECs in the G0/G1 phase, resulting in the increased cells in the G0G1 phase and decreased in the S phase (P<0.05). RT-PCR showed that NO-Fluvastatin could obviously inhibit the CyclinE mRNA expression and induce the P21wafl mRNA expression as compared with the negative control groups (P<0.05). This experiment suggested that NO-Fluvastatin could suppress the proliferation of HLECs by regulating cell cycle regulatory proteins (inhibiting the expression of CyclinE mRNA and inducing the expression of P21wafl mRNA), resulting in the arrest of HLECs in the G0/G1 phase, which can offer theory basis for NO-Fluvastatin in treating posterior capsular opacification in clinic practice.

d that sodium salicylate can induce the expression of HSP27 in HLECs-B3. The effects are mediated, at least in part, through the activation of P38MAPK and ERK1/2 signaling pathway.

The diagnosis and treatment of the lacrimal passage obstruction with lacrimal endoscope was investigated and its subsidiary surgical procedures were evaluated. Ninety-three patients (109 eyes) with lacrimal passage obstruction, including presaccal canalicular obstruction (PSCO) and na-solacrimal duct obstruction (NLDO), were examined under a lacrimal endoscope, and the obstruc-tions were treated with laser or micro-drill. All patients were followed up after the operation for 3-6 months. The difference between the laser and the micro-drill treatment was observed. During the pe-riod of follow-up, the curative rate was 82.57%. The healing rate in PSCO group and NLDO was 80.36% and 84.91% respectively (P>0.05). After treatment with the laser, the healing rate was 93.33% in the PSCO group and 66.67% in the NLDO group respectively (P<0.05). After treatment with the micro-drill, the healing rate in PSCO and NLDO groups was 65.39% and 94.28% respec-tively (P<0.01). The lacrimal passage obstruction can be observed and treated directly through the lacrimal endoscope. Choosing different surgical procedures in operation according to the locations of the obstruction is helpful to improve the effectiveness.

Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kipl-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endo- thelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combi- nation was used as negative control (pGenesil-HK). The recombination of four plamids was con- firmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kipl was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kipl mRNA and p27Kipl protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group), pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The pro- liferation rates of the pGenesil-P3 group, the pGenesii-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kipl on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesiI-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kipl could down-regulate the expression of p27Kipl effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.