To explore the potential mechanism of molecular hydrogen in the regulation of miRNA expression and signal-modulating activities.
Retinal microglia cells were activated by Lipopolysaccharides (LPS) and then treated with hydrogen-saturated medium or normal medium without hydrogen. qRT-PCR was used to detect the expression difference in miR-9, miR-21 and miR-199 between these two groups. Moreover, the expression of LPS-induced signaling proteins, including Myd88, IKK-β, NF-κB, and PDCD4, were detected by Western blotting.
The results demonstrated a marked down-regulation of miR-9 and miR-21 and up-regulation of miR-199 by hydrogen treatment; the expression of Myd88 and IKK-β was decreased after hydrogen treatment, whereas PDCD4 was increased, and there was no significant change in NF-κB expression.
The results in the present study indicate that miR-9, miR-199 and miR-21 play an important role in the anti-inflammatory regulation of LPS-activated microglia cells by molecular hydrogen, which will help to explain the protective mechanism of molecular hydrogen against inflammatory injury.
hydrogen; Lipopolysaccharides; miR-9; miR-21; miR-199; Toll-like receptor 4
To report the results of combined vitrectomy, lensectomy and silicone oil (SO) tamponade in treating primary rhegmatogenous retinal detachment (RRD) associated with choroidal detachment (CD).
A retrospective, consecutive and case series study of 21 subjects with concurrent RRD associated with CD was conducted. All subjects underwent a standard three-port 20G pars plana vitrectomy (PPV) with lensectomy and silicone oil tamponade. Mean follow-up time was 8 months (rang from 4 to 19 months). The primary and final anatomic success rate, visual acuity and final intraocular pressure(IOP) were recorded and analyzed.
Of 21 subjects, 8 were women and 13 were men. Age at presentation ranged from 22 to 75 years (mean 57.4 years). The presenting vision ranged from light perception to 0.15. The initial IOP ranged from 3mmHg to 12mmHg (mean 6.2mmHg). All eyes were phakic except one pseudophakic. No intraocular lens was implanted during the primary surgical intervention. Fifteen of 21 (71.4%) eyes had retina reattached after one operation. Six eyes had recurrent inferior retinal detachment due to proliferation. Five of them were successfully reattached after one or more additional operations. Mean IOP at final follow-up was 15.2mmHg (range from 8mmHg to 20mmHg). One case declined for further operation. The final reattachment rate was 95.2%. Visual acuity improved in 19 (90.5%) eyes, was unchanged in 1 (4.8%) eye and decreased in 1 (4.8%) eye.
Combination of vitrectomy, lensectomy and silicone tamponade is an effective method in treating RRD associated with CD, reducing the incidence of postoperative hypotony.
choroidal detachment; lensectomy; rhegamatogenous retinal detachment; vitrectomy; silicone oil
To investigate inhibitory γ-aminobutyric acid (GABA) ergic postsynaptic currents (IPSCs) and postsynaptic currents (PSCs) in layer IV of the rat visual cortex during the critical period and when plasticity was extended through dissolution of the perineuronal nets (PNNs).
We employed 24 normal Long-Evans rats to study GABAA-PSC characteristics of neurons within layer IV of the visual cortex during development. The animals were divided into six groups of four rats according to ages at recording: PW3 (P21-23d), PW4 (P28-30d), PW5 (P35-37d), PW6 (P42-44d), PW7 (P49-51d), and PW8 (56-58d). An additional 24 chondroitin sulfate proteoglycan (CSPG) degradation rats (also Long-Evans) were generated by making a pattern of injections of chondroitinase ABC (chABC) into the visual cortex 1 week prior to recording at PW3, PW4, PW5, PW6, PW7, and PW8. Immunohistochemistry was used to identify the effect of chABC injection on CSPGs. PSCs were detected with whole-cell patch recordings, and GABAA receptor-mediated IPSCs were pharmacologically isolated.
IPSC peak current showed a strong rise in the age-matched control group, peaked at PW5 and were maintained at a roughly constant value thereafter. Although there was a small increase in peak current for the chABC group with age, the peak currents continued to decrease with the delayed highest value at PW6, resulting in significantly different week-by-week comparison with normal development. IPSC decay time continued to increase until PW7 in the control group, while those in the chABC group were maintained at a stable level after an initial increase at PW4. Compared with normal rats, the decay times recorded in the chABC rats were always shorter, which differed significantly at each age. We did not observe any differences in IPSC properties between the age-matched control and penicillinase (P-ase) group. However, the change in IPSCs after chABC treatment was not reflected in the total PSCs or in basic membrane properties in layer IV of the rat visual cortex.
Our results demonstrate that rather than rapidly increasing during the critical period for neuronal plasticity, IPSCs in layer IV of rat visual cortex are maintained at an immature level when PNNs are removed by chABC. This suggests that GABA receptor maturation involves the conformation of the CSPGs in PNNs.
gamma-aminobutyric acid receptor; plasticity; visual cortex; development; postsynaptic currents; chondroitinase ABC; chondroitin sulfate proteoglycans; whole-cell patch recording
To determine whether the PI3K/AKT/mTOR pathway is activated in proliferative vitreoretinopathy (PVR) in homo-sapiens.
The retina of controls and patients with PVR were collected and their levels of PI3K, phospho-AKT, phospho-mTOR, phospho-p70S6k and phospho-4EBP-1 were determined by Western blot. The cultured human retinal pigment epithelial cell line D407 was treated with a specific mTOR inhibitor, rapamycin (RAPA) or a PI3K inhibitor, LY294002, of various concentrations and durations. Cell morphology was observed by phase contrast microscopy and the proliferation and apoptosis of treated cells were determined by MTT assay and flow cytometry.
Levels of PI3K, phospho-AKT, phospho-mTOR, phospho-P70S6K and phospho-4EBP1 was increased in the retina in PVR (P<0.05). In D407 cells, both RAPA and LY294002 significantly inhibited cell proliferation and cell cycle progression, and promoted apoptosis (P <0.05); morphologically, the cells became smaller. Both RAPA and LY294002 reduced levels of phospho-AKT, phospho-mTOR, phospho-p70S6k and phospho-4EBP1 expression (P <0.05). RAPA, but not LY294002, had no significant effect on PI3K expression.
PI3K/AKT/mTOR signaling pathway is highly activated in the retinal pigment epithelial cells of PVR. The inhibitors of PI3K/AKT/mTOR signaling pathway, RAPA and LY294002, could inhibited the PI3K/AKT/mTOR signaling pathway by reducing the levels of phosphorylation of mTOR pathway components.
human retinal pigment epithelial cell; proliferative vitreoretinopathy; PI3K/AKT/mTOR signal pathway
To study clinical features and gene mutations within the paired-like homeodomain transcription factor 2 (PITX2) gene in a pedigree of bilateral limbal dermoids.
Complete eye examinations have been performed on each individual of the family. Exons of paired-like homeodomain transcription factor 2 (PITX2) were amplified by polymerase chain reaction, sequenced, and compared with a reference database.
We described the phenotype, clinic findings in a family with two affected members. The masses of the proband's eyes were excised surgically demonstrating a dermoid cyst by histopathological examination. No mutation was detected in the gene PITX2 in this pedigree.
A family of limbal dermoid cyst was reported. In addition, no pathogenic sequence variations were found in PITX2, indicating that this phenotype in this family is a distinctive entity.
Pedigree; dermoid cyst; limbus; PITX2 gene
To present retinal microstructure, metabolism and function abnormalities in the course of multiple evanescent white dot syndrome (MEWDS) by Heidelberg spectralis modality imaging platform and observe its outcome by EDI-SD-OCT and two wavelength autofluorescence.
A case of multiple evanescent white dot syndrome in a 23-year-old female presented initially with a 15-day history of floaters and a central scotoma in the right eye. To establish the diagnosis, multimodality imaging was performed, namely, blue light-fundus autofluorescence (BL-FAF, excitation 488nm, emission >500nm), near-infrared fundus autofluorescence (NIR-FAF, excitation 787nm, emission >800nm) using a confocal scanning laser ophthalmoscope, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA), spectrum-domain enhance depth imaging optical coherence tomography (SD-EDI-OCT), multifocal electroretinography (mf-ERG) and fundus photogragh were performed and followed up at the eighth month after initially visiting.
Optical coherence tomography (OCT) showed a transient disruption of the foveal photoreceptor outer segments in correspondence to foveal granularity. NIR-FAF showed hypoautofluorescent areas, ≤40µm in size, mostly concentrated around the posterior pole and its temporal side less than that in BL-FAF. Mf-ERG show pinnacle disappeared in fovea and macula and responses decreased markedly compared with the follow eye. At the eighth month follow up, hyperfluorescence in BL-FAF were disappear, while, NIR-FAF Hypofluorescent spots in early stage of such lesion were reduced. But OCT demonstrated the structure was recovered in residual Hypofluorescent area in NIR-FAF. The subfoveal choroidal thickness was decreased from 372µm to 307µm slightly and cost line was recovered.
MEWDS is a benign self-healing disease and there is no pathological evidence to investigate the natural course of such disease. SD-OCT allows highly detailed images approaching histopathology to certify the microstructural changes. Two-wave length FAF and mf-ERG provide more information about metabolism in outer retina especial RPE and photoreceptor. Spectralis OCT combined with two-wavelength FAF and mf-ERG provide a new way to analyze this disease and offer more details for therapy and follow-up.
MEWDS; Spectralis OCT; NIR-FAF; BL-FAF; mf-ERG
Wavefront aberration affects the quality of retinal image directly. This paper reviews the representation and reconstruction of wavefront aberration, as well as the construction of virtual eye model based on Zernike polynomial coefficients. In addition, the promising prospect of virtual eye model is emphasized.
wavefront aberration; wavefront reconstruction; virtual eye; Zernike polynomial
To detect the expression of transforming growth factor beta-induced gene (TGFBI) protein in human corneal tissue and overexpress it in the human corneal epithelial cells in order to discuss the function of TGFBI in the pathogenesis of corneal dystrophy.
Immunohistochemistry (IHC) was used to detect the expression of TGFBI in the human cornea tissue. TGFBI cDNA was obtained by reverse transcription-PCR from human corneal total RNA extracted from cornea transplant donor and cloned into pCMV-N-HA vector. The recombinant pCMV-N-HA-TGFBI plasmid transfected human corneal epithelial cells. Forty-eight hours later, mRNA and proteins were harvested from cells for real-time PCR analysis and western blot assay respectively.
IHC indicated TGFBI mainly exist below the human corneal epithelium layer. Transfection of recombinant pCMV-N-HA-TGFBI into human corneal epithelial cells resulted in effective expression of TGFBI, as shown by increased mRNA level detected by real-time PCR as well as increased protein level detected by Western blot. Meanwhile the result of real-time PCR and Western blot shown the expression of MMP1, MMP3 (matrix metalloproteinases MMP) increased while the expressin of TIMP1 (tissue inhibitors of matrix metalloproteinases TIMP) decreased.
TGFBI mainly exists below the corneal epithelial layer, recombinant eukaryotic expression vector harboring human TGFBI cDNA was obtained and efficiently overexpressed in human corneal epithelial cells. Meanwhile the TGFBI overexpression in human corneal epithelial cells result in MMP1, MMP3 increasing and TIMP1 decreasing. The result might be helpful for studying the function and role of TGFBI in pathogenesis of corneal dystrophy.
TGFBI; human corneal epithelial cells; matrix metalloproteinases
The prevalence of diabetic retinopathy (DR) is increasing dramatically as the population of patients with diabetes continues to rise. This paper wants to investigate the prevalence and risk factors of DR in China through reviewing the research from Pubmed about population-based epidemic studies. The results of observational studies suggested that the overall prevalence of DR was 1.6%-6.5% in population, 19.9%-43.1% in diabetes and 13.6% in population without diabetes, most of the DR were the mild type, macular edema and vision-threatening were 5.2% and 1.2%. The risk factors for DR were longer duration of diabetes, plasma glucose concentration, concentrations of HbA1c level, higher systolic blood pressure(BP), higher diastolic BP, male gender, rural region, and methods of diabetic treatment and so on. The prevalence of DR which was strongly related to duration of diabetes was higher with the increase of diabetes. DR would be the major leading cause of visual impairment in China, it is very important to prevent DR by early screening and any other methods.
diabetic retinopathy; prevalence; prevention; risk factors
To quantify the changes in the lens profile with accommodation in different age groups.
The Pentacam HR system was used to obtain the images of the anterior eye segment from 23 young and 15 presbyopic emmetropic subjects in unaccommodated (with an accommodation stimulus of 0.0D) and accommodated (with an accommodation stimulus of 5.0D for the young group and 1.0D for the presbyopic group) states. The phakic crystalline lens shape, including curvature of crystalline lens and central lens thickness (CLT), and the measurements of anterior segment length (ASL), central anterior chamber depth (CACD) were investigated. The anterior chamber volume (ACV) was also measured.
The reduction of CACD and ACV were significant in both groups after accommodation stimulus. From the profile of anterior eye segment, a significant decrease in anterior crystalline lens radii of curvature (-2.52mm) and a mean increase in CLT (0.222mm) and ASL (0.108mm) were found in the young group with an accommodation stimulus of 5.0D. However, no statistically significant changes of CLT, ASL, or crystalline lens radii of curvature were found in the presbyopic group.
Our data showed that the shallowing of anterior chamber during accommodation was caused by the forward bulging of the anterior lens surface, rather than by anterior shifting of lens position in either young or presbyopic subjects.
accommodation; crystalline lens; Pentacam HR; anterior eye segment
To analyze the incidence and risk factors for retinopathy of prematurity (ROP).
A retrospective analysis was conducted on 568 premature infants from September 2005 to December 2010 with birth weight(BW) equal to or less than 2 500g or a gestational age(GA) at birth of 34 weeks or less. All of the members were examined by indirect binocular ophthalmoscopy.
ROP occurred with an incidence rate of 10.7% among 568 premature infants, and stages 3 and above ROP occurred with an incidence rate of 2.5%. This study showed the infants were more prone to develop ROP with short geststional age, low BW, long time of oxygen inhalation, and severe infants diseases. Twins had a significantly higher rate of ROP(18.3%) than singleton babies(9.8%), ROP were severer in twins than singleton babies.
Short GA, low BW, long time of oxygen inhalation, severe infants diseases, and non-singleton babies were the most significant risk factors associated with ROP.
retinopathy of prematurity; ROP epidemiology; ROP incidence
To describe the design and preliminary results of the hospital based epidemiological study for diabetic retinopathy(HBESDR), an ongoing epidemiological study to estimate the prevalence of diabetic retinopathy(DR) and to elucidate the clinical, anthropometric, biochemical and any other risk factors associated with diabetic retinopathy.
Totally 2000 diabetes will be recruited from the Diabetes eye clinic in the First Affiliated Hospital of China Medical University. All subjects underwent blood sugar estimation and Oral Glucose Tolerance Test to diagnose diabetes. All diabetes would undergo complete questionnaire, a comprehensive eye examination. Blood and urine would be collected for biochemical investigations. All fundus photographs for any DR will be graded. Participants who need treatment will be sent to the ophthalmic clinic and follow-up interval program for all subjects will be suggested. A computerized database is created for the records.
To date, 1174 diabetes have been recruited, there were 350(29.81%) DR in all diabetes, most of them were with mild non-proliferative diabetic retinopathy (NPDR) (139, 39.71%); 71(20.29%) moderate NPDR, 66(18.86%) severe NPDR, 74(21.14%) proliferative diabetic retinopathy (PDR). Females, longer duration of diabetes, family history of diabetes and hypertension had a statistically significant increase in risk of any DR.
The study is expected to provide an estimate of the overall prevalence of DR and the prevalence with different duration of diabetes and also a better understanding of the risk factors associated with DR.
diabetic retinopathy; prevalence; epidemiolo-gical study
To investigate the frequency of idiopathic phacodonesis (IP) in senile cataract subjects and the short-term clinical outcomes following cataract surgery.
This institutional case-control study included 1301 consecutive low-income cataract subjects from June to November 2009. Anterior segment were carefully evaluated with dilated pupil under slit-lamp. IP were screened and graded by a criteria set by the authors. Risk factors, surgical outcomes, and operative complications were analyzed.
A total of 42 subjects (3.2%) with IP were diagnosed and classified as grade 1 (36 subjects), grade 2 (5 subjects) and grade 3 (1 subject). Harder lenses and intumescent cataracts were observed in the IP group than the control group (P<0.05). Logistics regression test also indicated the main risk factor was the hardness of the lens. The incidence of zonular dialysis during surgery was 23.8% (10 eyes), which was significantly higher than the controls (0.7%, P<0.001). Visual outcomes of the two groups were not statistically or clinically significant.
Hard nucleus and intumescent cataract are related to IP in senile cataract subjects in Qinghai, China. With more care being taken, grade 1 and some of the grade 2 IP subjects achieved similar surgical outcomes as compared to controls.
phacodonesis; lens nucleus; zonular dialysis
To analyze osteopontin (OPN) expression in vitreous and proliferative retinal membranes of patients with proliferative vitreous retinopathy (PVR).
A total of 54 vitreous fluid samples were obtained between 2009 and 2010, which contained 45 with PVR (group A) and 9 without PVR (group B). Enzyme-linked immunosorbent assay was applied to quantify the OPN concentrations in vitreous fluid. Four samples of proliferative retinal membrane were also obtained at the time of vitrectomy, and their contents of OPN were measured by Real-time RT-PCR.
The OPN levels in the vitreous fluid were 778.48±62.06ng/mL in group A and 452.99±32.52ng/mL in group B. The vitreous OPN levels in group A were significantly higher than those in group B and to rise by time in the early stages of PVR. The average OPN levels in the proliferative retinal membranes (F=0.14) were also higher than those in the retinal pigment cells (F=0) using Real-time RT-PCR.
The high vitreous and proliferative retinal membrane OPN levels in PVR suggest that OPN might promote the development of PVR. The vitreous OPN concentrations are rising by the time in the early phases of PVR.
osteopontin; proliferative vitreous retinopathy; vitreous; proliferative retinal membranes
To estimate the incidence of cilioretinal arteries among Han population of north China.
A cross-sectional sample of subjects aged from 20 years old to 80 years old were selected using a cluster sampling technique from Shenyang of Liaoning Province for Diabetic Eye Disease Study. Eligible subjects were recruited in the Community Health Center which took the stereo fundus photography using 45°Non-Mydriatic Fundus Camera. Data were analyzed by SPSS 14.0.
The incidence of cilioretinal arteries in 5000 eyes of 2500 subjects was determined. One, two or more cilioretinal arteries were present in 876 of all subjects and in 923 of all the eyes. Difference of presence, number and distribution of cilioretinal arteries was observed. During the subjects, the arteries occurred bilaterally in 6.9% and contributed to some portion of the temporal circulation in 78.3%, nasal circulation 14.8%, respectively. Statistical analysis indicated that there was no significant difference in incidence between men and women, left and right eyes (P>0.05).
35.0% persons have the cilioretinal arteries in Han population of north China. Men and women have an equal distribution of cilioretinal arteries.
cilioretinal artery; incidence; population
To study the differences of fibroblast growth factor receptor 1 (FGFR1) gene on human lens epithelial cells (HLECs) of adults and fetuses.
Indirect in situ RT-PCR was adopted for detection of FGFR1 gene. The cDNA of the mRNA in the paraffin sections of fetus and adult HLEC was synthesized by reverse transcription reaction. After PCR amplification, in situ hybridization test was performed with synthesized oligonucleotide probe and relative quantification was carried out using image analysis.
HLECs of adults and fetuses expressed FGFR1 gene, the expression level was higher in fetuses than in adults. The difference between them had significance (P<0.05).
FGFR1 exist in HLEC and the expression is age-related, which could be one of causes of the high occurrence of post operational after-cataract in children.
human lens epithelial cells; fibroblast growth factor receptor 1; indirect in situ RT-PCR; after-cataract
To estimate the effects of human umbilical vein (HUV) implanted under the sclera of glaucoma model on intraocular pressure (IOP) lowering and to investigate its related mechanisms
A total of 20 human umbilical veins (HUV) were collected from healthy fetus umbilical core. After the establishment of glaucoma model in rabbits, human freeze-dried umbilical vein was implanted under the sclera during NPDS, while for control group, sclerostomy was performed without implant. The formation of the filtration bleb and IOP were detected every 24 hours before surgery and on day 3, 7, 10 and 14 after surgery. Handheld pen-type Tono-pen II tonometer was used to measure IOP after topical anesthesia treatment. Each measurement has three duplicates. The incision recovery, filtration, conjunctiva congestion and anterior chamber inflammation were observed everyday after surgery.
IOP was decreased dramatically with less inflammation than traditional sclerostomies with the application of HUV. The significant differences of IOP between the NPDS with and without HUV implant groups were shown up from 10 days after surgery. The average IOP in NPDS without HUV implant was 14.25mmHg, while for NPDS with HUV implant group, it was 12.30mmHg. This structure of filtration bleb, which allowed the aqueous humor to leave the eye, was formed for any type of surgery. However, 1-2 weeks later, filtration bleb was still existed in the group of sclerostomy with HUV implant and more stable than that of the surgery without HUV implant. Histological observations were performed on day 3, 7 and 14 after surgery. For the eyes under sclerostomy with HUV implant, HUV lumina was shown up on 3 days after surgery with few fibroblast cells near the sclera. On 7 days after surgery, HUV lumina was stably maintained but with obvious fibroblast cells and inflammatory cell. On 14 days after surgery, HUV lumina was still clearly observed but with scarring formation, which suggests that the IOP lowering effects might result from an effective drainage structure formation.
HUV might be an alternative material to make the drainage pathway for non-penetrating deep sclerostomy.
IOP-lowering; non-penetrating deep sclerostomy; human umbilical vein
To detect the effect of connective tissue growth factor (CTGF) on the apoptosis in the diabetic retina with small interfering RNAs (siRNA) targeting CTGF.
A total of 60 rats were divided into 6 groups including control group, diabetic 4, 8, 12, 16 weeks groups, and interference group. Diabetic rats were induced by intraperitoneal streptozotocin (STZ). Retinas were obtained from control, diabetic rats and diabetic rats of interference group treated by intravitreal injection of CTGFsiRNA to suppress the expression of CTGF mRNA. Retinal cells apoptosis was detected by Tunnel staining and mRNA expression of CTGF was analyzed by RT-PCR.
The levels of CTGF and the apoptosis in the retinas of diabetic rats were significantly higher than those in the controls. Apoptosis occurred at 4 weeks after a diabetic model being set up, became serious with the diabetes developing, while CTGF elevated at 8 weeks. The apoptosis cell counts increased to 25.8cells/mm2 at 24weeks of diabetes. SiRNA-mediated inhibition of CTGF mRNA resulted in a significant decrease in apoptosis. Significant correlations were found between CTGF and apoptosis in the retina.
It was suggested that CTGF might be involved in retinal cells apoptosis which is a characteristic of early diabetic retina. SiRNA targeting CTGF seems to have the advantage of ameliorating retinal cells apoptosis.
apoptosis; CTGF; retina; diabetes
To observe effect of adenovirus-mediated brain derived neurotrophic factor in early retinal neuropathy of diabetes in rats.
Adult male Wistar rats, 9 weeks of age, were injected intraperitoneally with STZ to induce diabetes. Two weeks after the models were established, Ad.BDNF was administered into the vitreous cavities of rats. Four weeks after the models were set up, the rats were killed and the retina was removed for Western blotting and whole-mount immunohistochemistry for tyrosine hydroxylase (TH) to observe the changes of TH and dopaminergic amacrine cells in retina.
The protein levels of TH and the number of positive staining dopaminergic amacrine cells and the staining gray scale of experimental group without Ad.BDNF were lower statistically. But there was no statistically significant difference between the experimental group with Ad.BDNF and control group.
In the early stage of STZ diabetic, the administration of Ad.BDNF into the vitreous cavities can increase TH protein levels and the density of dopaminergic amacrine cells in the STZ rats' retina.
diabetic retinopathy; tyrosine hydroxylase; brain derived neurotrophic factor; adenovirus
To study the differences of basic fibroblast growth factor (bFGF) gene expression in lens epithelial cells (LECs) between fetuses and cataract patients.
In situ hybridization was used to detect bFGF mRNA in the LECs that were cultured and in tissue sections from fetuses and in the LECs from the anterior capsule of cataract patients. Image analysis was used for the relative quantitative analysis of bFGF mRNA.
bFGF gene existed in the LECs that were cultured and in tissue sections from fetuses and in the LECs from the anterior capsule of cataract patients. The integral absorbance for the fetal cultured cells, the fetal tissue sections and the capsule membrane cells of cataract patients were 627.1±268.7, 131.5±42.8 and 79.2±26.3 respectively. The integral absorbance of fetal cultured LECs was significantly higher than that of fetal section LECs (P<0.01). The integral absorbance of cataract LECs was significantly lower than that of fetal LECs (P<0.01).
The in vitro culture of LECs can improve bFGF gene expression. The bFGF gene expression in fetal LECs is significantly higher than that in cataract LECs.
lens epithelial cells; environmental scanning electron microscope; in situ hybridization; basic fibroblast growth factor; image analysis
To explore the effects and potential mechanisms of curcumin on retinal Müller cell in early diabetic rats.
Diabetic rats were induced by a single intraperitoneal injection of streptozotocin (STZ). Male Sprague-Dawley (SD) rats were randomly assigned into 4 groups: control group (naïve SD rats administered with a single intraperitoneal injection of citric buffer), diabetic group (STZ-diabetic rats), dimethyl sulfoxide (DMSO) group (diabetic rats intraperitoneally administered with mixture of DMSO and normal saline, once a day) and curcumin group (diabetic rats intraperitoneally administered with curcumin, 80mg/kg, once a day). Three months after diabetes onset, malondialdehyde (MDA, indication of oxidative stress level) and reduced glutathione (GSH) in retina were detected with kits, glial fibrillary acidic protein (GFAP) in retina was revealed by immunohistochemistry and Western blot, and retinal glutamine synthetase (GS) were observed by Western blot.
Compared with control group, retinal MDA was increased, and GSH was decreased in diabetic and DMSO groups (P<0.05, respectively). While, retinal MDA and GSH in curcumin group showed no difference compared with control group (P>0.05). Furthermore, up-regulation of retinal GFAP and down-regulation of retinal GS were detected in diabetic and DMSO groups, and no alteration could be observed in curcumin group revealed with Western blot. Compared with control group, retinal Müller cells showed significant increase in GFAP immunochemistry staining in diabetic and DMSO groups. Moreover, GFAP-positive staining was decreased in curcumin group compared with diabetic group.
Curcumin inhibits diabetic retinal oxidative stress, protects Müller cell, and prevents the down-regulation of GS in diabetic retina. Therefore, curcumin has a therapeutic potential in the treatment of diabetic retinopathy (DR).
diabetic retinopathy; curcumin; oxidative stress; Müller cell
To reconstruct the lamellar cornea using human amniotic epithelial (HAE) cells and rabbit cornea stroma in vitro using tissue engineering technology.
Human amnia taken from uncomplicated caesarean sections were digested by collagenase to obtain HAE cells, and the cells were cultured to proliferate. Rabbit corneal epithelial cells were removed by n-heptanol to make lamellar matrix sheets. The second passage of HAE cells were cultured on the corneal stroma sheets for 1 or 2 days, then transferred to an air-liquid interface environment to culture for 2 weeks. Tissue engineered lamellar cornea (TELC) morphology was observed by Hematoxylin-eosin (HE) staining; its ultrastructure was observed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM); corneal epithelial cell-specific keratin 3 and keratin 12 were detected with immunofluorescence microscopy.
HAE cells grew on the rabbit corneal stroma, forming a monolayer after 1-2 days. About 4-5 layers of epithelial cells developed after 2 weeks of air-liquid interface cultivation, a result similar to normal corneal epithelium. Rabbit corneal stromal cells were significantly reduced after one week, then almost completely disappeared after 2 weeks. TEM showed desmosomes between the epithelial cells; hemidesmosomes formed between the epithelial cells and the basement membrane. SEM revealed that the HAE cells which grew on the lamellar cornea had abundant microvilli. The tissue-engineered cornea expressed keratin 3 and keratin 12, as detected by immunofluorescence assay.
Functional tissue-engineered lamellar corneal grafts can be constructed in vitro using HAE cells and rabbit corneal stroma.
amniotic epithelial cells; cornea; tissue engineering; keratin
To screen mutations in the retinitis pigmentosa 1 (RP1) gene and the rhodopsin (RHO) gene in Chinese patients with retinitis pigmentosa sine pigmento (RPSP) and describe the genotype-phenotype relationship of the mutations.
Twenty affected, unrelated Chinese individuals with RPSP (4 autosomal dominant RPSP, 12 autosomal recessive RPSP and 4 unknown inheritance pattern) were recruited between 2009 and 2012. The clinical features were determined by complete ophthalmologic examinations. Polymerase chain reaction (PCR) and direct DNA sequencing were used to screen the entire coding region and splice junctions of the RP1 gene and the RHO gene. The cosegregation analysis and population frequency studies were performed for patients with identified mutations.
Five variants in the RP1 gene and one in the RHO gene were detected in 20 probands. Four missense changes (rs444772, rs446227, rs414352, rs441800) and one non-coding variant (rs56340615) were common SNPs and none of them showed a significant relationship with RPSP. A missense mutation p.R1443W was identified in the RP1 gene in three affected individuals from a family with autosomal dominant RPSP and was found to cosegregate with the phenotype in this family, suggestive of pathogenic. In addition, population frequency analysis showed the p.R1443W mutation was absent in 300 healthy controls.
The identification of p.R1443W mutation cosegregating in a family with autosomal dominant RPSP highlights an atypical phenotype of the RP1 gene mutation, while RHO gene is not associated with the pathogenesis of RPSP in this study. To our knowledge, this is the fist mutation identified to associate with RPSP.
retinitis pigmentosa sine pigmento; RP1 and RHO gene; gene mutation
To observe effects of trabeculectomy with amniotic membrane transplantation (AMT) in controlling postoperative intraocular pressure (IOP) in patients with medically uncontrolled glaucoma.
This study included adult patients with requiring bilateral glaucoma surgery. Each patient underwent trabeculectomy (Non-AMT group) in one eye and with AMT (AMT group) in the other eye according to randomized principle. Success was defined as intraocular pressure (IOP)<21mmHg without any anti-glaucoma medications at 24 months follow-up. The two groups were compared in terms of IOP, complications and success rate.
Thirty-four eyes of 17 patients were investigated in this study. There was no statistically signifcant difference in pre-operative IOP between the two groups. The mean IOP was lower in AMT group compared with Non-AMT group on follow up months 12, 18, and 24.Postoperative complications were more frequent in Non-AMT group (35.3%, 6/17) compared with AMT group (5.9%, 1/17). The success rate of surgery was 88.2% (15/17) in Non-AMT group and 100% (17/17) in AMT group.
Trabeculectomy with AMT is an effective procedure to reduce IOP and complications, thereby improving surgical success rates.
trabeculectomy; amniotic membrane transplantation; glaucoma