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Problem The purpose of the study was to investigate the different expressions of toll-like receptors (TLRs) in the proliferative and the secretory phase of endometrial tissue. Method of study Eight infertile women were included in this study. The endometrial tissues of proliferative and secretory phase were obtained from each woman. The tissues were evaluated for the expression of mRNA for TLR1–10 by reverse transcribed polymerase chain reaction (RT-PCR). The proteins of TLR2, 3, 4, and 9 were evaluated by Western blot. The mRNA and protein level of proliferative and secretory endometrial tissue from the same woman was compared. The data were analysed with SPSS15.0. Results TLR1–7, 9, and 10 mRNA were expressed throughout the menstrual cycle, but in the same woman, the expression of TLR2–6, 9, and 10 mRNA was higher during the secretory phase than that in the proliferative phase. The Western blot also showed that the protein expression of TLR2, 3, 4, and 9 was stronger in the secretory phase than that in the proliferative phase in the same woman. Conclusion The expression of TLRs is cycle dependent in human endometrial tissue. The expressions of TLRs were higher in the secretory phase than that in the proliferative phase: this indicated that TLRs may be regulated by sex hormones throughout the menstrual cycle.

To investigate the expression of toll-like receptors (TLRs) in the human cornea with Fusarium solani (F. solani) keratitis.Five human corneas with F. solani keratitis and 5 healthy human corneas were evaluated for TLR1-10 mRNA by reverse transcribed polymerase chain reaction (RT-PCR) and relative real-time PCR. The proteins of TLR2, 4, and 9 were also compared by Western blot. The mean times between these corneas were collected, and the onset of treatment to surgery was 34.4 +/- 12.4 days, ranging from 15 to 47 days. The data were analyzed with SPSS15.0.TLR1-10 mRNA was expressed in both healthy and F. solani infected human corneas. The expression of TLR2, 4, 6, and 9 mRNA in the F. solani infected human corneas were upregulated. The Western blot showed the protein expression of TLR2, 4, and 9 was also upregulated in the corneas with F. solani keratitis than that of the healthy corneas.TLRs are expressed diversely in the F. solani infected human cornea, and TLR2, 4, and 9 may be implicated in the pathogenesis of F. solani infection in the cornea.

Purpose Neutrophils release neutrophil extracellular traps (NETs) when defending against invading microorganisms. We investigated the existence of NETs in fungal keratitis. Methods Fourteen patients with unilateral fungal keratitis were included. Detailed information about each patient was recorded, including (1) patient history (onset of symptoms and previous therapy), (2) ocular examination findings by slit-lamp biomicroscopy, (3) laboratory findings from direct smear examination and culture of corneal scrapings, (4) NET formation, and (5) treatment strategy and prognosis. Immunofluorescence staining was used to evaluate the existence of NETs on corneal scrapings. The relationship between the quantification of NETs and the clinical character of the fungal keratitis was identified. Results NETs were identified in all 14 patients. Patients with a higher grade of NET formation and fewer fungal hyphae always showed a good treatment response and a short course of infection. NETs were consistently found mixed with fungal hyphae in the corneal scrapings from infected patients. No statistical significance was found between the grade of NETs formed and the course of infection before presentation, and no relationship between the quantification of NETs and the size of the ulcer was found. Conclusions The results suggest that NETs are involved in fungal keratitis. The number of NETs in infected corneas may provide a tool for evaluating the prognosis for fungal keratitis.

Purpose. It has been confirmed that inflammatory cytokines are involved in the progression of pterygium. Histamine can enhance proliferation and migration of many cells. Therefore, we intend to investigate the proliferative and migratory effects of histamine on primary culture of human pterygium fibroblasts (HPFs). Methods. Pterygium and conjunctiva samples were obtained from surgery, and toluidine blue staining was used to identify mast cells. 3-[4, 5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) was performed to evaluate the proliferative rate of HPFs and human conjunctival fibroblasts (HCFs); ki67 expression was also measured by immunofluorescence analysis. Histamine receptor-1 (H1R) antagonist (Diphenhydramine Hydrochloride) and histamine receptor-2 (H2R) antagonist (Nizatidine) were added to figure out which receptor was involved. Wound healing model was used to evaluate the migratory ability of HPFs. Results. The numbers of total mast cells and degranulated mast cells were both higher in pterygium than in conjunctiva. Histamine had a proliferative effect on both HPFs and HCFs, the effective concentration (10 μmol/L) on HPFs was lower than on HCFs (100 μmol/L), and the effect could be blocked by H1R antagonist. Histamine showed no migratory effect on HPFs. Conclusion. Histamine may play an important role in the proliferation of HPFs and act through H1R.

To investigate the relationship between meibomian gland dysfunction (MGD) and incomplete blinking caused by cranial nerve seven (CN VII) palsy.A prospective case series of 60 consecutive patients with unilateral CN VII palsy was evaluated for MGD. According to the House-Brackmann scale, patients were divided into complete or incomplete blinking group, with the incomplete group further subdivided to early, middle or late stage according to the paralysis duration. Schirmer's I test, tear break-up time (BUT), superficial punctate keratopathy, eyelid abnormality, ability and quality of meibum expression were evaluated. Unaffected contralateral eyes were used as control for comparison.A paired sample t-test between affected and unaffected eyes demonstrated a significant difference for BUT, superficial punctate keratopathy, eyelid abnormality, ability and quality of meibum expression in the incomplete blinking group. However, only BUT showed significant difference between affected and unaffected eyes in complete blinking group. Furthermore, we showed that paralysis duration was related to the incidence of MGD. Significant difference was demonstrated between the affected and unaffected eyes for superficial punctate keratopathy, eyelid abnormality, ability and quality of meibum expression in the middle and late stage, but not early stage.These findings suggest that the inability to blink completely induced by CN VII palsy for more than one week might contribute to the development of MGD. Clinical optimization of blinking may ameliorate MGD symptom and benefit CN VII palsy patients.

To evaluate the use of lenticules extracted using small incision lenticule extraction (SMILE) surgery as a surgical alternative for wound closure in corneal perforation.Corneal lenticules obtained through SMILE surgery with central thickness100 μm were fixed over corneal perforation sites using 10-0 nylon interrupted stitches. Patients were monitored for a minimum of 1 year and were assessed using slit-lamp microscopy, fluorescein, digital tonometry, and best spectacle-corrected visual acuity (BSCVA) measurements. Postoperative complications throughout the study period were recorded.Corneal perforations were successfully sealed in all 6 patients; 3 patients (50%) exhibited improved postoperative BSCVA. Between 3 and 4 weeks after the operation, part of the lenticules became incorporated into the corneal stroma and complete reepithelialization was achieved. During the follow-up period of 12 months, no evidence of infection, relapse, or perforation was detected in any patient.These preliminary findings suggest that the use of corneal lenticules may be a safe and effective surgical alternative for corneal perforation closure, with potential clinical application as relatively simple and inexpensive temporary measures to improve the condition of the cornea for further definitive interventions.

Since the 21st century, the development of corneal tissue engineering technology has been developing rapidly. With the progress of biomaterials, cell culture and tissue engineering technology, tissue engineering cornea has gained great development in both basic scientific research and clinical application. In particular, tissue engineered corneal scaffolds are the core components of tissue engineered corneas. It is the focus of current research on tissue engineering cornea to search for scaffolds with good biocompatibility, high safety and good biomechanical properties. In this paper, the recent research progress of tissue engineering corneal materials is reviewed.

PurposeDexmedetomidine can prolong the duration of local anesthetics, but the effect of retrobulbar dexmedetomidine on the potency of ropivacaine for retrobulbar block has not been investigated. Our study was designed to determine the effect of retrobulbar dexmedetomidine on ropivacaine for retrobulbar block in children.MethodsA group of 90 children aged 10-16 years scheduled for vitreoretinal surgery who received retrobulbar block were randomly assigned to 1 of 3 groups: group L (retrobulbar ropivacaine), group LD1 (ropivacaine plus 0.5 µg.kg−1dexmedetomidine), or group LD2 (ropivacaine plus 1 µg.kg−1dexmedetomidine). The minimum local anesthetic concentration (MLAC) was determined according to a Dixon-Massey protocol. The primary endpoint of the study was MLAC. Secondary outcomes were duration of postoperative analgesia, postoperative pain scores, dexmedetomidine side effects, and time to hospital discharge.ResultsThe MLAC values of retrobulbar ropivacaine were 0.314%, 0.259%, and 0.246% in groups L, LD1, and LD2, respectively. The median (interquartile range) durations of analgesia in the postoperative period were 66 (54-117), 89 (40-157), and 168 (120-194) minutes in groups L, LD1, and LD2, respectively (L vs LD1 or LD2, pConclusionsRetrobulbar dexmedetomidine reduces the MLAC values of ropivacaine and improves postoperative analgesia in children without any neurologic side effects.

Purpose The pathological mechanisms underlying cataract formation remain largely unknown on account of the lack of appropriate in vitro cellular models. The aim of this study is to develop a stable in vitro system for human lens regeneration using pluripotent stem cells. Methods Isolated human urinary cells were infected with four Yamanaka factors to generate urinary human induced pluripotent stem cells (UiPSCs), which were induced to differentiate into lens progenitor cells and lentoid bodies (LBs). The expression of lens-specific markers was examined by real-time PCR, immunostaining, and Western blotting. The structure and magnifying ability of LBs were investigated using transmission electron microscopy and observing the magnification of the letter "X," respectively. Results We developed a "fried egg" differentiation method to generate functional LBs from UiPSCs. The UiPSC-derived LBs exhibited crystalline lens-like morphology and a transparent structure and expressed lens-specific markers αA-, αB-, β-, and γ-crystallin and MIP. During LB differentiation, the placodal markers SIX1, EYA1, DLX3, PAX6, and the specific early lens markers SOX1, PROX1, FOXE3, αA-, and αB-crystallin were observed at certain time points. Microscopic examination revealed the presence of lens epithelial cells adjacent to the lens capsule as well as both immature and mature fiber-like cells. Optical analysis further demonstrated the magnifying ability (1.7×) of the LBs generated from UiPSCs. Conclusions Our study provides the first evidence toward generating functional LBs from UiPSCs, thereby establishing an in vitro system that can be used to study human lens development and cataractogenesis and perhaps even be useful for drug screening.

Meibomian gland dysfunction (MGD) is believed to be the leading cause of dry eye (DE) disease worldwide. The connection between aging and MGD has long been recognized. However, few studies have addressed the relationship between MGD and hormone replacement therapy (HRT) in perimenopausal women, and not have examined the prevalence of MGD in perimenopausal women. The purpose of this study was to address and evaluate the possible relationship between MGD and HRT in perimenopausal women. The results suggest that perimenopausal women have a high prevalence of DE related to perimenopausal symptoms. The study also shows that perimenopausal women who use HRT can gain benefits for DE as well as for perimenopausal symptoms. Physicians caring for women who are experiencing DE related to perimenopausal symptoms should consider HRT.

Purpose The aim of this study was to investigate hyaluronic acid (HA) protection on cultured human corneal epithelial cells (HCEs) against benzalkonium chloride (BAC)-induced DNA damage and intracellular reactive oxygen species (ROS) increase. Methods Cells were incubated with different concentrations of BAC with or without the presence of 0.2% HA for 30 min. DNA damage to HCEs was examined by alkaline comet assay and by immunofluorescence microscopic detection of the phosphorylated form of histone variant H2AX (γH2AX) foci. ROS production was assessed by the fluorescent probe, 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Cell apoptosis was determined with annexin V staining by flow cytometry. Results HA significantly reduced BAC-induced DNA damage as indicated by the tail length (TL) and tail moment (TM) of alkaline comet assay and by γH2AX foci formation, respectively. Moreover, HA significantly decreased BAC-induced ROS increase and cell apoptosis. However, exposure to HA alone did not produce any significant change in DNA damage, ROS generation, or cell apoptosis. Conclusions BAC could induce DNA damage and cell apoptosis in HCEs, probably through increasing oxidative stress. Furthermore, HA was an effective protective agent that had antioxidant properties and could decrease DNA damage and cell apoptosis induced by BAC.

To investigate the toxic effects of benzalkonium chloride (BAC), a preservative commonly used in ophthalmic preparations, on DNA single- and double-strand breaks in immortalized human corneal epithelial cells (HCEs). HCEs were treated with BAC in concentrations ranging from 0.00005% to 0.001% for 30 min. Cells were examined immediately after BAC exposure and after 24-h recovery. Alkaline comet assay was used to detect DNA single-strand breaks (SSBs). Immunofluorescence microscope detection of the phosphorylated form of histone variant H2AX (γH2AX) foci indicated DNA double-strand breaks (DSBs). Cell viability was measured by the MTT test. A significant increase of SSBs, detected by alkaline comet assay, was observed in a dose-dependent manner with BAC exposure in HCEs at concentrations of 0.00005% and higher. Such BAC treatment also exhibited a dose-dependent increase in DSBs, evaluated by number of γH2AX foci. In addition, a significant change in the relative cell survival rate of HCEs was observed after exposure to 0.001% BAC for 30 min. Although the toxic effects of BAC could be partly repaired after 24 h of cell recovery, SSBs and DSBs in HCEs were still present after BAC removal. The results demonstrated that exposure to BAC in HCEs, even at low concentrations, could induce DNA strand breaks, which were present after BAC removal. Cell survival analysis indicated that BAC-induced DNA damage was correlated with the cytotoxic effects.

Aim: We report retrospectively on the change of the axial length of three high myopia patients who accept bilateral phacoemulsifi cation, but delayed their second eye operation more than 25 months. Methods: All the three observed patients delayed their second eye operation over 25month. The follow up time was more than 24 months after the second eye surgery. The measurements such as axial length and lens opacity were assessed. Results: The second eye operation was delayed 25, 33 and 48 months. The clinical observation showed the exacerbation of the cataract as the time going on. The axial length of delayed surgery eye was changed from 26.09, 26.28, 26.21 mm to 27.05, 26.85, 26.72 mm, but the fellow eyes was changed from 25.81, 31.48, 26.05 mm to 25.76, 31.63, 26.16 mm. The axial length was almost unchanged after the next follow up. Conclusion: Cataract may affect the axial length of high myopes in the adults. Physicians should be aware of the possible development of axial length in cataract patients with myopia.

The purpose of this study was to investigate hydrocortisone's potential in inhibiting the expression of VEGF in cultured human cornea fibroblasts (HCF) and to determine whether the role of hydrocortisone in the expression of VEGF may relate to toll-like receptors 2 and 4 (TLR2 and TLR4).Three different concentrations of hydrocortisone were used to stimulate the cultured HCF. The cellular abundance of the mRNAs for VEGF and TLR2, 4 were determined by real-time polymerase chain reaction analysis. The release of IL-6, IL-8, and VEGF from cultured HCF was measured using enzyme-linked immunosorbent assays (ELISA) in the presence and absence of specific blocking antibodies to TLR2, 4. The proteins of TLR2, 4 were also compared by Western blot.Following incubation of HCF with hydrocortisone, we found the mRNA expression of TLR2, 4 and VEGF were markedly inhibited. ELISA and Western blot analysis confirmed that protein expression of TLR2, 4 and VEGF was down-regulated in response to hydrocortisone. The result of ELISA also showed the release of IL-6 and IL-8 can be inhibited by hydrocortisone. But all these inhibitions were partly counteracted after pretreatment with anti-TLR2 and/or anti-TLR4 monoclonal antibody.Hydrocortisone may decrease the expression of VEGF through inhibiting TLR2, 4 activity in cultured human corneal fibroblasts.

Purpose To evaluate the effect of glucocorticoids on the expression and function of Toll-like receptors (TLRs) in human corneal fibroblasts (HCFs). Methods Cultured HCF cells were stimulated with three different concentrations of hydrocortisone. The effect on the expression of TLR2 and TLR4 was determined by real-time PCR. The TLR2, TLR4, and pIκB-α proteins were compared by western blot. The release of IL-6 and IL-8 was measured using enzyme-linked immunosorbent assay in the presence and absence of TLR2 and TLR4-specific blocking antibodies. Results Incubation of HCFs with hydrocortisone markedly inhibited the expression of TLR2 and TLR4 mRNAs and decreased the release of IL-6 and IL-8 in a dose-dependent manner. Western blot analysis confirmed that expression of TLR2, TLR4, and pIκB-α was also downregulated in response to hydrocortisone. The result of ELISA also showed the release of IL-6 and IL-8 can also be inhibited by hydrocortisone. However, all these inhibitions were counteracted after pretreatment with anti-TLR2 and anti-TLR4 monoclonal antibodies. Conclusions Glucocorticoids, such as hydrocortisone, can inhibit the expression of TLR2 and TLR4 on HCFs, and thus may increase susceptibility to cornea infections. Our results suggest that topical glucocorticoids may affect the cornea’s innate immunity through TLRs.

The purpose of the study was to investigate the different expressions of toll-like receptors (TLRs) in the proliferative and the secretory phase of endometrial tissue.Eight infertile women were included in this study. The endometrial tissues of proliferative and secretory phase were obtained from each woman. The tissues were evaluated for the expression of mRNA for TLR1-10 by reverse transcribed polymerase chain reaction (RT-PCR). The proteins of TLR2, 3, 4, and 9 were evaluated by Western blot. The mRNA and protein level of proliferative and secretory endometrial tissue from the same woman was compared. The data were analysed with SPSS15.0.TLR1-7, 9, and 10 mRNA were expressed throughout the menstrual cycle, but in the same woman, the expression of TLR2-6, 9, and 10 mRNA was higher during the secretory phase than that in the proliferative phase. The Western blot also showed that the protein expression of TLR2, 3, 4, and 9 was stronger in the secretory phase than that in the proliferative phase in the same woman.The expression of TLRs is cycle dependent in human endometrial tissue. The expressions of TLRs were higher in the secretory phase than that in the proliferative phase: this indicated that TLRs may be regulated by sex hormones throughout the menstrual cycle.

PURPOSE To study the expression of toll-like receptors (TLRs) in healthy corneas and corneas with active or nonactive herpetic stromal keratitis (HSK). METHODS Eight corneas with HSK (including 5 active and 3 nonactive cases) and 8 healthy corneas were evaluated for TLR1-10 mRNA by reverse transcription-polymerase chain reaction (RT-PCR) and relative real-time PCR. The proteins of TLR2, 9 in 3 different groups, were also compared by immunofluorescence staining. The data were analyzed with SPSS 11.5. RESULTS TLR1-10 mRNA was expressed in both healthy and HSK corneas. In the healthy cornea, the mRNA expression of TLR1, 2, 3, 4, and 6 was higher, whereas the expression of TLR7, 8, and 9 was lower relative to TLR 5 and 10. All the TLRs mRNA expression in the active HSK corneas were upregulated, especially TLR4, 7, 8, and 9, compared with the healthy corneas. However, in nonactive HSK corneas, only TLR7 mRNA expression upregulated, whereas the others downregulated. The immunofluorescence staining showed that the expression of TLR 9 was slightly stronger in the cornea with active HSK than that of nonactive and healthy corneas. CONCLUSIONS TLRs are expressed diversely in the healthy cornea and could have an important role in innate corneal immunity. TLR4, 8, and 9 may be implicated in the pathogenesis of active HSV infection in the cornea, whereas TLR7 may play a key role in HSK whether it is active or not.

Purpose To evaluate the role of toll-like receptors (TLRs) in host responses to Fusarium solani by the use of cultured immortalized human corneal epithelial cells (HCEC) and to determine whether inactive hyphal fragments can induce an antifungal response in these cells. Methods Cultured HCEC cells were stimulated with inactive hyphal fragments from Fusarium solani, and the effect on expression of TLRs was determined by real-time polymerase chain reaction (PCR), immunofluorescence, and western blot analysis. Cells were also cocultured with hyphal fragment and hydrocortisone to determine whether hydrocortisone modulates the transcription of TLRs. The release of interleukin-6 (IL-6) and IL-8 was also measured using enzyme-linked immunosorbent assays (ELISA) in the presence and absence of specific blocking antibodies to TLR2 and TLR4. Results Incubation of HCECs with inactive hyphal fragments upregulated the expression of TLR2, 3, 4, and 6 mRNAs and increased the release of IL-6 and IL-8. Immunofluorescence staining and western blot analysis confirmed that expression of TLR2 and TLR4 was upregulated in response to hyphal fragments. This upregulation was further enhanced by cotreatment with hydrocortisone. Results from ELISA assays showed that the concentration of IL-6 was increased, and the concentration of IL-8 was decreased in supernatants of HCECs after treatment with both hydrocortisone and hyphal fragments. The release of IL-6 and IL-8 was also inhibited by incubation with anti-TLR2 and anti-TLR4 monoclonal antibodies. Conclusions HCECs are involved in the cornea immune response to fungal infections. TLR2 and TLR4 may play a crucial signaling role in response to Fusarium hyphae in HCECs. Glucocorticoids such as hydrocortisone can enhance the expression of the TLRs on the epithelium and thus may enhance the resistance to fungal infections in the cornea. These findings may provide crucial information for understanding the immune mechanisms of fungal keratitis and promote the design of new immune therapeutical approaches to fungal keratitis.

To study the safety and effectiveness of a combination of both intravenous injection of lidocaine and intraocular pressure-lowering medications, in the intraocular pressure control and relief of symptoms of refractory acute primary angle-closure glaucoma (PACG).Five consecutive patients with their first attack of acute PACG, with intraocular pressureor = 45 mmHg and a failure to release from the attack under antiglaucomatous medications for 4 hours, were recruited into the study. On presentation, each patient received topical pilocarpine and timolol, and systemic acetazolamide and mannitol as primary treatment. Then the patients accepted 2% lidocaine by intravenous injection at dose of 0.8 mg/kg in concert with antiglaucomatous medications. The intraocular pressures after intravenous injection at 30 minutes, and then at 1, 2, 4, 12, and 24 hours, were documented by applanation tonometry. Symptoms, visual acuity, intraocular pressure, corneal edema, angle status on gonioscopy, pupillary size, and reaction were also measured.Six eyes of five patients seen with acute PACG were recruited. The mean intraocular pressure was reduced from 50.83 +/- 5.34 mmHg to 39.5 +/- 3.45 mmHg at 30 minutes after intravenous injection, and then to 33.3 +/- 3.56 mmHg at 1 hour, and 24.55 +/- 5.09 mmHg at 2 hours after intravenous injection. The mean intraocular pressure was less than 21 mmHg at 4 hours and beyond. There was instant symptomatic relief for all patients. No complications were encountered.From this preliminary study, intravenous injection of lidocaine seems to be safe and effective in controlling intraocular pressure and eliminating symptoms in acute PACG. But the exact efficacy and safety need further investigation in large case studies.

Background Treatment of lacrimal sac abscess of the traditional surgical approach may result in complications from cutaneous fistula formation, damage the sac, cause skin scarring and even have the potential for inducing cicatricial ectropion. We designed a new treatment scheme that is expected to achieve internal drainage with the use of lacrimal probe and crawford silicon tube. Methods A prospective study was performed for the management of lacrimal sac abscesses. All suitable patients from January 2011 to June 2014 were managed by lacrimal probe and crawford tube insertion. Postoperatively, patients received 0.5% Levofloxacin eye drops four times per day and oral Levofloxacin tablets 0.5 g once per day for four days. Follow-up times were for more than three months after removing the Crawford tube. The condition of the lacrimal sac and the patient’s symptoms were carefully evaluated. Results Fourteen patients suffering from lacrimal sac abscesses were included in this study. A history of chronic dacryocystitis was found in six patients, after acute dacryocystitis was found in three patients, and nasolacrimal occlusion with epiphora was found in other five patients. Resolution of signs and symptoms of lacrimal sac abscesses in all fourteen patients. No recurrence of lacrimal sac abscesses occurred during the median follow up period of 20.9 ± 7.8 months (range 6–36 months). Epiphora reoccurred in four patients. Conclusions Lacrimal probe and crawford silicon tube is successful as a procedure of choice for lacrimal sac abscesses. The insertion of a Crawford tube also offers potential advantages over standard treatment with the lack of recurrence of dacryocystitis or infection in post-surgical patients.