Evaluation value of subjective visual quality examination on surgical indications of the early cataracts based on objective scatter index values.

Aim: To evaluate the subjective visual functions of early cataracts patients and assess their surgical indications. Methods: Eyes were separated into a control group (Group A without cataract) and two early cataracts groups (Group B with 2.0 ≤ OSI < 3.0 and Group C with 3.0 ≤ OSI < 4.0). The objective scatter index (OSI), modulation transfer function cut-off frequency (MTF cut-off), and Strehl ratio (SR) values were applied to measure objective visual functions. The contrast sensitivity (CS) and scores of the questionnaires (QOL and VF-14) characterized subjective visual functions. Above visual functions were compared among three groups. Postoperative visual functions in Group B and C were analyzed to assess the outcome of surgery. Results: Ninety two subjects (126 eyes) were included in the study. All objective visual function in Group B were significantly better than Group C (all P < 0.01), but worse than Group A (all P < 0.01). Except for 1.5 c/d CS, subjective visual function in Group A were significantly better than Group B and C (all P < 0.05), but there was no significant differences between Group B and C. As for eyes that underwent surgery in Group B and C, all visual functions significantly improved after surgery (P < 0.05), except for 1.5 c/d CS in Group C. There were no significant differences among the three groups after surgery. Conclusion: The subjective visual function can be impaired in early cataracts patients with OSI < 3.0, whose objective visual functions were statistically better than patients with OSI ≥ 3.0. These patients can benefit equally from surgery as patients with OSI ≥ 3.0. Subjective visual functions can be used as surgical indications for these patients.

Changes in Meibum Lipid Composition With Ocular Demodex Infestation.

Purpose: The purpose of this study was to understand the impact of Demodex infection in the lipid component of meibum in patients. Methods: The meibum samples were collected from four groups of subjects: (1) Demodex-negative with non-MGD (D-M-; n = 10); (2) Demodex-positive with non-MGD (D+M-; n = 10); (3) Demodex-negative with MGD (D-M+; n = 10); and (4) Demodex-positive with MGD (D+M+; n = 10). A liquid chromatography-mass spectrometry (LC-MS) system consisting of ultra-performance liquid chromatography and a Q Exactive high-resolution mass spectrometer was used for lipids separation and detection. Results: Compared with the D-M- group, the D+M- group had lower levels of phosphatidylcholines (PCs) and lysophosphatidylcholines (LPCs) and higher levels of phosphatidylethanolamines (PEs). Compared with the D-M+ group, the levels of sphingomyelins (SMs) and PCs in the D+M+ group were decreased, whereas the levels of (O-acyl)-ω-hydroxy fatty acids (OAHFAs), ceramides (CERs), LPCs, and diacylglycerols (DGs) were significantly increased. Triacylglycerols (TGs), DGs, CERs, and OAHFAs were decreased in D-M+ group, whereas levels of PEs, phosphatidylinositols, and phosphatidylglycerols were increased in meibum obtained from the D-M+ group compared with those in the D-M- group. TGs, SMs, CERs, and PEs were decreased in the D+M+ group, whereas levels of LPCs, LPEs, PCs, and PEs were increased in meibum from the D-M+ group compared with those in the D+M- group. Conclusions: To the best of our knowledge, this is the first study to assess the changes in meibum from patients with ocular Demodex infestation. The significant increase of OAHFAs in the Demodex-positive group suggest that OAHFAs may be associated with the progress of ocular Demodex infections. Translational Relevance: OAHFAs could be a potential new therapeutic target for ocular Demodex infestation.

Hyperkeratinization and Proinflammatory Cytokine Expression in Meibomian Glands Induced by Staphylococcus aureus.

Purpose: This exploratory study aimed to investigate the morphological and pathological alterations of the meibomian gland (MG) with the Staphylococcus aureus crude extracts (SACEs) treatment. Methods: Mouse MG explants were cultured and differentiated with or without SACEs for 48 hours. Explant's viability and cell death were determined by thiazolyl blue tetrazolium bromide (MTT) assay and TUNEL assay. MG morphology was observed by Hematoxylin and Eosin staining. Lipid droplet production was detected by Nile Red staining and LipidTox immunostaining. The pro-inflammatory cytokines were detected by ELISA. The relative gene and protein expression in MG explants was determined via quantitative RT-PCR, immunostaining, and immunoblotting. The components of the SACEs were analyzed by immunoblotting and silver staining. Results: Our findings demonstrated that the SACEs treatment induced overexpression of keratin 1 (Krt1) in the ducts and acini of MG explants, accompanied by a decrease in viability and an increase in cell death in explants. Furthermore, the SACEs treatment dose-dependently increased the levels of TNF-α, IL-1ß, and IL-6 in MG explants. The SACEs treatment induced activation of the nuclear factor kappa B (NF-κB) and AIM2 (absent in melanoma 2)/ASC (apoptosis-associated speck-like protein containing a caspase recruitment domain) inflammasome signaling pathway in explants. Further investigation showed expression of the key adipogenesis-related molecule peroxisome proliferator-activated receptor γ was decreased after SACEs treatment. However, no change was found in the lipid synthesis of MG explants after treatment with the SACEs. Staphylococcal enterotoxins B (SEB) was detected in the SACEs. SEB induced the overexpression of Krt1 and IL-1ß in ducts and acini of MG explants. Conclusions: Our findings confirm that Staphylococcus aureus induced hyperkeratinization and pro-inflammatory cytokines expression in MG explants ducts and acini. These effects might be mediated by SEB. Activation of the NF-κB and AIM2/ASC signaling pathway is involved in this process.

Real dynamic assessment of tear film optical quality for monitoring and early prevention of dry eye.

To evaluate real dynamic assessment of tear film optical quality for monitoring and prevention of dry eye.Right eyes of 62 normal and 39 dry eye subjects were included. Dynamic measurement of objective scatter index (OSI) was performed by using the Optical Quality Analysis System II (OQAS II), correlation coefficient between OSI and time (CCOT) was calculated. According to whether the CCOT was significantly ascending, normal and dry eye groups were further subdivided for comparison. By using Scheimpflug-Placido topographer, non-invasive tear break-up time (NITBUT) was recorded, and a 2-dimensional precorneal tear film map was reconstructed and divided into central, middle, and peripheral corneal zones, distribution of tear break-up spots in the 3 corneal zones were analyzed.The numbers of tear break-up spots were higher in all the 3 corneal zones of the dry eye subjects (P < .01), when compared with the normal subjects. The Dry Eye subjects with ascending CCOT had the shortest NITBUT (P < .001-.034) and the most tear break-up spots over the whole cornea (P < .001-.044). Between the dry eye subjects with non-ascending CCOT and those with ascending CCOT, difference of tear break-up spots was found significant only in the peripheral corneal zone (P < .01).Non-ascending and ascending CCOT of dry eye patients reflect different stability of tear film. Real dynamic assessment of tear film optical quality is potential for monitoring and early prevention of dry eye.

Organotypic Culture of Mouse Meibomian Gland: A Novel Model to Study Meibomian Gland Dysfunction In Vitro.

Purpose: Meibomian glands are essential in maintaining the integrity and health of the ocular surface. Meibomian gland dysfunction (MGD), mainly induced by ductal occlusion, is considered as the major cause of dry eye disease. In this study, a novel in vitro model was established for investigating the role of inflammation in the process of MGD. Methods: Mouse tarsal plates were removed from eyelids after dissection and explants were cultured during various time ranging from 24 to 120 hours. Meibomian gland epithelial cells were further enzymatically digested and dissociated from tarsal plates before culturing. Both explants and cells were incubated in different media with or without serum or azithromycin (AZM). Furthermore, explants were treated with IL-1ß or vehicle for 48 hours. Analyses for tissue viability, histology, biomarker expression, and lipid accumulation were performed with hematoxylin and eosin (H&E) staining, immunofluorescence staining, and Western blot. Results: Higher viability was preserved when explants were cultured on Matrigel with immediate addition of culture medium. The viability, morphology, biomarker expression, and function of meibomian glands were preserved in explants cultured for up to 72 hours. Lipid accumulation and peroxisome proliferator-activated receptor γ (PPARγ) expression increased in both explants and cells cultured in media containing serum or AZM. Treatment with IL-1ß induced overexpression of Keratin (Krt) 1 in meibomian gland ducts. Conclusions: Intervention with pro-inflammatory cytokine IL-1ß induces hyperkeratinization in meibomian gland ducts in vitro. This novel organotypic culture model can be used for investigating the mechanism of MGD.

Intense Pulsed Light Therapy for Patients with Meibomian Gland Dysfunction and Ocular Demodex Infestation.

To observe the clinical changes of meibomian gland dysfunctipn (MGD) and ocular Demodex infestation after intense pulsed light (IPL) treatment to further examine the mechanism of IPL treating patients with MGD and ocular Demodex infestation. The medical records of 25 patients (49 eyes) with MGD treated with IPL, were retrospectively examined to determine outcomes. Associated ocular-surface parameters (ocular surface disease index, OSDI; lipid layer thickness, LLT; noninvasive first breakup time, NIF-BUT; noninvasive average breakup time, NIAvg-BUT; tear film breakup area, TBUA; Schirmer I Test, SIT; corneal fluorescein staining, CFS), eyelid margin abnormalities, meibum quality and expressibility, MG morphological parameters (macrostructure and microstructure), and the number of Demodex infestation were examined before and after treatment. The MG microstructure and the Demodex infestation were examined via in vivo confocal microscopy (IVCM). The results showed that there were statistically significant differences in associated ocular-surface parameters (all P<0.05) before and after IPL treatment, except SIT (P=0.065). Eyelid margin abnormalities, meibum quality and expressibility obviously improved in upper and lower eyelid after IPL treatment (all P<0.0001). MG macrostructure (MG dropouts) decreased in upper (P=0.002) and lower eyelid (P=0.001) after IPL treatment. The nine parameters of MG microstructure in upper and lower eyelid all distinctly improved after IPL treatment (all P<0.0001). The mean number of Demodex mites on the upper lid margin (6.59±7.16 to 3.12±3.81/9 eyelashes) and lower lid margin (2.55±2.11 to 1.29±1.53/9 eyelashes) significantly reduced after IPL treatment (all P<0.0001). The Demodex eradication rate was 20% (8/40) in upper lid margin and 34.15% (14/41) in lower lid margin. These findings indicate that IPL shows great therapeutic potential for patients of MGD and ocular Demodex infestation.

The correlation between the microstructure of meibomian glands and ocular Demodex infestation: A retrospective case-control study in a Chinese population.

Meibomian gland dysfunction (MGD) is a common disease in ophthalmic clinic. This study aimed to explore ocular Demodex infestation on the microstructure changes of the meibomian glands (MGs) in patients with MGD by in vivo confocal microscopy (IVCM).We retrospectively reviewed 103 eyes of 52 patients with MGD and 62 eyes of 31 non-MGD patients. All enrolled patients underwent IVCM examination. The following IVCM parameters were recorded: the MG acinar density (MAD), MG acinar longest diameter (MALD), MG acinar shortest diameter (MASD), MG orifice area (MOA), severity of MG fibrosis (MF), MG acinar irregularity (MAI), meibum secretion reflectivity (MSR), inhomogeneous appearance of walls of acinar units (AWI) and periglandular interstices of acinar units (API), and the number of Demodex.The positive rate of Demodex infestation in MGDs was 89.32%, and statistically higher than control group (controls; P < .001). All parameters showed statistically significant differences between MGDs and controls (P < .001), and Demodex-negative group and Demodex-positive group (P < .05) in both MGDs and controls, except MAD (P = .826) in controls. The number of Demodex was positively correlated with MALD, MASD, MF, MAI, MSR, AWI, and API in MGDs and controls (P < .05), and negatively correlated with MAD and MOA in MGDs (P < .05). MOA showed a strong significant correlation with the number of Demodex in controls (P < .001), whereas there was no significant difference between the number of Demodex and the MAD in controls (P = .448).Demodex can cause microstructural changes of MGs, which can cause or aggravate MGD, and the more the number of Demodex infestation, the more serious the structural damage.

Computer modeling of drug delivery in the anterior human eye after subconjunctival and episcleral implantation.

Recently, subconjunctival and episcleral implants have been proposed in the treatment of anterior eye diseases. In order to improve the delivery efficacy, it is important to understand the transport process of the implanted drugs. A 3D computational model, which includes heat transfer, aqueous humor (AH) flow, as well as diffusive and convective transport of the drug concentration, is developed to study the temporal and spatial evolution of the drug in the anterior segment of a human eye after subconjunctival and episcleral implantation, with a focus on drug delivery to three targets: iris, lens, and trabecular meshwork (TM). The release rate of the implanted drug is based on experimental data and effects of implantation location, eye orientation, and AH flow are investigated. Our numerical results indicate that subconjunctival implantation is more effective than episcleral implantation for drug delivery to all the three targets, and the accumulative amount of drug delivered to the three targets is larger in the horizontally-facing eye than in the up-facing eye. Implantation at the 12 o'clock circumferential position is the most effective for drug delivery to iris and lens, and the 3 o'clock position is the most effective for drug delivery to TM. This study may help to better understand the delivery process of implanted drugs in the anterior human eye, and improve delivery efficacy for clinical treatment of anterior eye diseases.

Ocular demodicosis as a risk factor of pterygium recurrence.

PURPOSE: To evaluate ocular demodicosis as a potential risk factor in pterygium recurrence. DESIGN: Cross-sectional study to correlate clinical findings with laboratory data. PARTICIPANTS: We retrospectively reviewed 94 patients (43 with primary and 51 with recurrent pterygia), among whom 68 patients received surgical correction, and prospectively enrolled another 23 pterygium patients and 14 nonpterygium controls for measuring the tear level of interleukin (IL)-17. METHODS: All patients had microscopically confirmed ocular demodicosis. Statistical correlations were analyzed among age, sex, aqueous tear deficiency, dry eye, ocular demodicosis, follow-up period, surgical outcome, and tear levels of IL-17 measured by enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: Correlation between ocular demodicosis or IL-17 levels and pterygium recurrence. RESULTS: Among 94 patients, ocular demodicosis was more prevalent in patients with recurrent pterygium than those with primary pterygium (P = 0.015). During follow-up of 16.5 ± 11.5 months, 68 postsurgical patients developed 7 corneal recurrences, which constituted 7.4% of primary and 12.2% of recurrent pterygium (P = 0.820). They also developed 8 conjunctival recurrences. Kaplan-Meier survival analysis showed combined (P = 0.000), corneal (P = 0.044), and conjunctival (P = 0.002) recurrence was significantly higher among patients with demodicosis than those without. Conjunctival recurrence occurred within 6 months in eyes without demodicosis but extended beyond 6 months in eyes with demodicosis. In 34 postsurgical patients with demodicosis, the mite count of 14 patients with recurrence was significantly higher than that of 20 without (P = 0.005). The IL-17 level was significantly higher in patients with either pterygium or demodicosis than controls (P = 0.049 and 0.040, respectively), and the IL-17 level was further elevated in patients with both pterygium and demodicosis (all P<0.05). CONCLUSIONS: Ocular demodicosis is a risk factor for pterygium recurrence, especially for conjunctival recurrence, presumably by perpetuating chronic inflammation mediated by T-helper (Th)17 lymphocytes.

Conjunctivochalasis interferes with tear flow from fornix to tear meniscus.

PURPOSE: To determine whether conjunctivochalasis (CCh) interferes with tear flow from the fornix to the tear meniscus and depletes the fornix tear reservoir. DESIGN: Comparative case series. PARTICIPANTS: The study group of 24 CCh patients (8 asymptomatic and 16 symptomatic), 9 of whom underwent operative correction, was compared with a control group of 13 normal subjects. METHODS: After instilling a 5-µl fluorescein drop into the inferior fornix, the inferior tear meniscus was depleted using a capillary tube. The tear meniscus height, with and without blinking, was recorded and calculated by video meniscometer from sequential captured images. MAIN OUTCOME MEASURES: The recovery rate of the original meniscus height was compared among groups at each time point after maximal depletion. RESULTS: The recovery rate of the tear meniscus was significantly slower in symptomatic than asymptomatic CCh patients when compared with normal subjects. Blinking 5 times facilitated such recovery in normal subjects and in asymptomatic CCh patients to the same extent as the normal, but not in symptomatic CCh patients. Deepening of the inferior fornix by removing degenerated Tenon's and reconstruction by cryopreserved amniotic membrane improved the recovery rate in symptomatic CCh patients to the same extent as normal subjects. CONCLUSIONS: The tear reservoir in the fornix rapidly replenishes the meniscus under normal circumstances. Conjunctivochalasis obliterates tears not only in the meniscus, but also in the reservoir, explaining how symptoms develop in CCh patients. Blinking is an effective compensatory mechanism to distinguish the severity of CCh. Surgical correction should not only restore the tear meniscus, but also deepen the fornix in CCh patients. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Identification of a novel homozygous nonsense mutation in EYS in a Chinese family with autosomal recessive retinitis pigmentosa.

BACKGROUND: Retinitis pigmentosa is the most important hereditary retinal degenerative disease, which has a high degree of clinical and genetic heterogeneity. More than half of all cases of retinitis pigmentosa are autosomal recessive (arRP), but the gene(s) causing arRP in most families has yet to be identified. The purpose of this study is to identify the genetic basis of severe arRP in a consanguineous Chinese family. METHODS: Linkage and haplotype analyses were used to define the chromosomal location of the pathogenic gene in the Chinese arRP family. Direct DNA sequence analysis of the entire coding region and exon-intron boundaries of EYS was used to determine the disease-causing mutation, and to demonstrate that the mutation co-segregates with the disease in the family. RESULTS: A single nucleotide substitution of G to T at nucleotide 5506 of EYS was identified in the Chinese arRP family. This change caused a substitution of a glutamic acid residue at codon 1,836 by a stop codon TAA (p.E1836X), and resulted in a premature truncated EYS protein with 1,835 amino acids. Three affected siblings in the family were homozygous for the p.E1836X mutation, while the other unaffected family members carried one mutant allele and one normal EYS allele. The nonsense mutation p.E1836X was not detected in 200 unrelated normal controls. CONCLUSIONS: The EYS gene is a recently identified disease-causing gene for retinitis pigmentosa, and encodes the orthologue of Drosophila spacemaker. To date, there are only eight mutations in EYS that have been identified to cause arRP. Here we report one novel homozygous nonsense mutation of EYS in a consanguineous Chinese arRP family. Our study represents the first independent confirmation that mutations in EYS cause arRP. Additionally, this is the first EYS mutation identified in the Chinese population.

[Observation on treatment of lacrimal passage obstruction under lacrimal endoscope].

OBJECTIVE: To evaluate the therapeutic efficacy of lacrimal endoscope treatment for lacrimal passage obstruction, and to compare the effectiveness of endoscopically controlled laser surgery and micro-drill surgery for lacrimal passage obstruction. METHODS: It was a prospective random controlled trial. Eighty nine patients (104 eyes) with lacrimal passage obstruction, including presacral canalicular obstruction (PSCO) and nasolacrimal duct obstruction (NLDO), were collected from September 2006 to December 2006 in Department of Ophthalmology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology. Patients were examined by endoscopy of the lacrimal drainage system under local anesthesia to detect the obstruction and changes of lacrimal mucous membrane. The obstructions were treated with laser or microdrill. Irrigation was performed to prove the recanalization of the lacrimal passage followed by injected ointment with 0.3% tobramycin and 0.1% dexamethasone into the lacrimal passage. All patients were followed up after the operation for 9-12 months. The difference between the laser and the microdrill treatment was observed. Chi-square test was used to evaluate the curative effect and complications differences between these two groups. RESULTS: The obstruction scene in the lacrimal passage of 89 patients could be observed effectively. All obstructions (104/104 eyes) were eliminated after the operation. Through the follow-up, the cure rate reached 78.85% (82/104 eyes). The cure rate of PSCO group and NLDO group, reached 77.78% (42/54 eyes) and 80.00% (40/50 eyes), respectively (chi2 = 0.077, P = 0.782). The cure rate of laser group and micro-drill group, was 80.43% (37/46 eyes) and 77.59% (45/58 eyes), respectively (chi2 = 0.125, P = 0.724). The cure rate of laser treatment was 89.66% (26/29 eyes) in the PSCO group and 64.71% (11/17 eyes) in the NLDO group (P = 0.040). The cure rate of micro-drill treatment was 64.00% (16/25 eyes) in the PSCO group and 87.88% (29/33 eyes) in the NLDO group (chi2 = 4.664, P = 0.031). Hemorrhage and palpebral edema occurred in 10.87% (5/46 eyes) and 4.35% (2/46 eyes) after laser treatment, respectively. Percentage of hemorrhage and palpebral edema after the micro-drill treatment was 55.17% (32/58 eyes) (compared to the laser group, chi2 = 21.969, P = 0.000) and 6.90% (4/58 eyes) (compared to the laser group, chi2 = 0.017, P = 0.896). CONCLUSIONS: Lacrimal passage obstruction can be observed and treated directly through the endoscopy of lacrimal drainage system. Choosing an appropriate surgical procedure according to the locations of the obstruction can be helpful for improving the effectiveness of the operation.

[Effect on the proliferation of bovine corneal endothelial cells by small hairpin RNA interference targeting p27Kip1].

OBJECTIVE: To clarify the proliferation of bovine corneal endothelial cells (bCEC) by interference with the recombinant plasmid of short hairpin RNA (shRNA) against p27Kip1, a kind of cyclin-dependent kinase inhibitor (CKI). METHODS: It was an experimental study. Three p27Kip1-shRNA template DNA sequences containing small hairpin structure were designed and synthesized as experimental groups. Plasmid expressing irrelevant shRNA with a random combination was used as negative shRNA. The products were inserted into the Pgensil-1 plasmid and the recombinant plasmid of Pgenesil-P1, Pgenesil-P2, Pgenesil-P3 and Pgenesil-HK were constructed. The recombinant plasmids were transfected into bCEC cells with liposome and a blank group. The expression of mRNA and protein of p27Kip1 was detected by RT-PCR and Western blot after stable transfection, and the plasmid with the best inhibitory effect was selected. The growth of the experimental group, Pgenesil-HK group and blank group were assessed by MTT. The influence of shRNA-p27Kip1 on bCEC cell cycle was deteceted by flow cytometry (FCM). All statistical analyses were performed using one-way ANOVA. RESULTS: Restrictive enzyme digestion and sequence analysis showed that four recombinant plamids were constructed successfully and the aim sequence was obtained. The expression of p27Kip1 mRNA and p27Kip1 protein of Pgenesil-P1 group, Pgenesil-P2 group and Pgenesil-P3 group were all lower than that in the control group, including blank group and negative siRNA group. The inhibitive rate of mRNA reached 32.71%, 67.76% and 80.28% (F = 453.102, P = 0.000 in each group) and the inhibitive rate of protein reached 29.27%, 64.73% and 76.13% (F = 75.385, P = 0.000 in each group) compared with the blank group. As the lowest expression among the three positive shRNA group, Pgenesil-P3 was selected for the next steps. There was no significant difference between blank group and negative Pgenesil-HK of the expression of p27Kip1 protein (P = 0.356) and the express of p27Kip1 mRNA (P = 0.246). Compared with the control group and the blank group, the growth of the bCEC transfected by Pgenesil-P3 was significantly promoted with increased cell percent of S-phrase (F = 334.957, P = 0.000) and decreased cell percent of G1-phrase (F = 134.224, P = 0.000). CONCLUSIONS: shRNA-p27Kip1 can down-regulate the expression of bCEC effectively and increase the growth of bCEC. shRNA-p27Kip1 RNA interference may be an effective method to promote the proliferation of CEC.

Effects of mitogen-activated protein kinase signal pathway on heat shock protein 27 expression in human lens epithelial cells exposed to sodium salicylate in vitro.

The roles of mitogen-activated protein kinase (MAPK) signal pathway in sodium salicylate-induced expression of heat shock protein 27 (HSP27) in human lens epithelial cells (HLECs-B3) in vitro were investigated. HLECs-B3 were incubated in the fresh media containing sodium salicylate at different concentrations for different durations, and allowed to be recovered in fresh medium without sodium salicylate for different durations with or without pretreatment with p38MAPK inhibitor (SB203580), ERK1/2 inhibitor (PD98059) and JNK/SAPK inhibitor (SP600125). The expression of P38MAPK, ERK1/2, JNK/SAPK, phosphorylated P38MAPK, phosphorylated ERK1/2, phosphorylated JNK/SAPK and HSP27 was detected by Western blot. The expression of HSP27 mRNA and protein was detected by RT-PCR and immunohistochemistry respectively. It was found there was only weak expression of HSP27 in normal HLECs. The expression of HSP27 was not detectable in HLECs-B3 that were exposed to sodium salicylate (55 mmol/L) for 1-5 h. It was indicated that recovery from sodium salicylate (>35 mmol/L) significantly increased the synthesis of HSP27. The expression of HSP27 was up-regulated in HLECs-B3 under sodium salicylate recovery for 3 h, reached the peak level for 6 h, and returned to the level of control cells by 24 h. Activation of P38MAPK from sodium salicylate stimulation occurred at 30th min, and increased significantly at 1st h, then declined and returned to baseline level at 3rd h under sodium salicylate recovery. Activation of ERK1/2 occurred at 1st h and reached the peak level at 6th h under sodium salicylate recovery. However, JNK/SAPK was inactivated by sodium salicylate. The expression of HSP27 could be down-regulated with the pretreatment of SB203580 and PD98059 jointly. It is concluded 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.

Effects of Mitogen-activated Protein Kinase Signal Pathway on Heat Shock Protein 27 Expression in Human Lens Epithelial Cells Exposed to Sodium Salicylate in vitro / 华中科技大学学报（医学）（英德文版）

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.

Inhibitory effects of NO-fluvastatin on proliferation of human lens epithelial cells in vitro by modulating cell cycle regulatory proteins.

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 P21(waf1) 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 mumol/L). Flow cytometry revealed that NO-Fluvastatin could significantly block HLECs in the G(0)/G(1) phase, resulting in the increased cells in the G(0)/G(1) 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 P21(waf1) 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 P21(waf1) mRNA), resulting in the arrest of HLECs in the G(0)/G(1) phase, which can offer theory basis for NO-Fluvastatin in treating posterior capsular opacification in clinic practice.

Effect of p27Kip1 inhibition on proliferation of bovine corneal endothelial cells by RNA interference.

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.

Inhibitory Effects of NO-Fluvastatin on Proliferation of Human Lens Epithelial Cells in vitro by Modulating Cell Cycle Regulatory Proteins / 华中科技大学学报（医学）（英德文版）

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.

Effect of p27Kipl Inhibition on Proliferation of Bovine Corneal Endothelial Cells by RNA Interference / 华中科技大学学报（医学）（英德文版）

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.

Diagnosis and treatment of lacrimal passage obstruction by lacrimal endoscope.

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.