Intravitreal anti-VEGF injection have been widely used in retinal vascular diseases and achieved good efficacy. Early pregnancy is an important period for fetal organ formation and vascular development. Studies have proved that VEGF plays an important role in maintaining the fetal and placental vascular system, and its loss or decline will affect embryonic development and lead to abortion. The use of intravitreal anti-VEGF during pregnancy is controversial, which may cause systemic side effects to the mother and fetus. This paper summarizes the literature of 23 cases on the use of anti-VEGF during pregnancy. Three cases reported loss of pregnancy with concomitant exposure to intravitreal bevacizumab, which suggested that we should be careful about the use of anti-VEGF during pregnancy and explain the possibility of ocular and systemic side effects to patients in detail. When deciding whether to use anti-VEGF, we should consider the relationship between exposure time and the critical period of vascular development and the systemic exposure of different drugs. Currently, there is a lack of large sample size studies on the use of anti-VEGF in pregnancy, and its safety needs to be further observed.
Objective To observe the effects of bevacizumab on aquaporin 4 (AQP4) expression in human retinal Muuml;ller cells in vitro under hypoxia. To explored the mechanism of treating retinal edema with bevacizumab. Methods Human Muuml;ller cells were cultured using the enzymatic digestion method. Transmission electron microscopic analysis and immunofluorescence staining identified Muuml;ller cells. With semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), the expression of AQP4 mRNA and vascular endothelial growth factor （VEGF） mRNA in Muuml;ller cells cultured under different concentration of COCl2 for different hours were observed. The expression of AQP4 mRNA in Muuml;ller cells cultured using CoCl2 precultured with 200 mu;g/ml bevacizumab was measured. Results More than 95% of primary cells showed positive reaction to glial fibrillary acidic protein, glutamine synthetase, vimentin and alpha;-smooth muscle actin with immunofluorescence staining. Characteristic 8-10 nm intracellular filaments could be seen in the cytoplasm viewed with transmission electron microscopy. The results using RT-PCR showed that CoCl2 increased the AQP4 and VEGF mRNA expression in Muuml;ller cells in a dose and time dependent manner (r=0.952, 0.954;P＜0.05). The expression of AQP4 mRNA in Muuml;ller cells was increased by VEGF (F=12.43,P＜0.05). The expression of AQP4 mRNA was significantly decreased by bevacizumab (F=2 370.37，P＜0.05). Conclusion Bevacizumab can down-regulate the expression of AQP4 mRNA in human Muuml;ller cells under hypoxic conditions partially by VEGF path, which may be a mechanism for treating retinal edema with bevacizumab.
Objective To evaluate the effect of vitreoretinal surgery on tear film.Methods Ninety-seven patients (97 eyes) of retinal detachment undergoing surgical therapy including vitrectomy, scleral buckling or scleral encircling were included in this study. Uncomfortable eye symptoms were inquired and tear break-up time (BUT), Schirmer Ⅰ test (SⅠt), corneal fluorescein staining (CFS), tear meniscus height (TMH) were measured at three days before surgery and at six time-points after surgery (two, 14 days and one, two, three, six months). Thirty patients were randomly chosen to receive impression cytology of conjunctiva at three days before surgery and at four time-points after surgery (one, two, three, six months).Results Comparing with the preoperative results, at the first three time-points after surgery (two, 14 days, one month), uncomfortable symptoms (t=-25.082,-9.966, -4.718,P＜0.01) and CFS scores(t=-6.244,-3.716,-4.683, P＜0.01) increased, tear breakup time (BUT) shortened greatly (t=9.960, 5.627, 4.953; P＜0.01). SⅠt and TMH increased significantly（t=-25.931,-5.839;-25.345,-3.873;P＜0.01） at two and 14 days after surgery. The number of conjunctival goblet cell decreased significantly at one month after surgery（t=2.259, P＜0.05).All those tear film parameters returned to preoperative level at two months after surgery.Conclusion Vitreoretinal surgery influences tear film stability transiently, and the tear film function tends to restore in two months after the operation.
Objective To investigate the early effects of intervention with tanakan on retinal function in diabetic retinopathy(DR) after laser photocoagulation. Methods Prospective random controlled study was performed on 60 Patients (60 eyes) from 23 to 69 years old with DR(phase Ⅲ～Ⅳ). The multifocal electroretinograms (MERG) were tested with VERIS Ⅳ before, the 3rd day and the 7th day after photocoagulation. Results No significant differences were found in the latencies and response densities of N1,P1 and N2 between the two groups before photocoagulation. Compared with that before photocoagulation, three days after photocoagulation the latencies in tanakan group had no significant change. The response densities of N1,P1 and N2 reduced and the changes were much smaller than that in control. Three days after photocoagulation, the response densities of P1 and N2 in the central macula 5°area were much higher and the latencies of P1 and N2 were significantly shorter than that in control group. There were no significant differences in the response densities in the 7th day and the differences in the latencies between two groups still existed. Conclusion Tanakan may be effective in preventing the retina from damage of retinal photocoagulation in some degree in DR. (Chin J Ocul Fundus Dis, 2002, 18: 208-211)
ObjectiveTo investigate the effects of thrombospondin-1 active fragment (TSP-1) synthetical peptide VR-10 on proliferation and migration of rhesus choroidal-retinal endothelial (RF/6A) cell and the expressions of apoptosis relative genes in RF/6A cell. MethodsThe survival rate of RF/6A cell were detected by methyl thiazolyl tetrazolium, and migration ability was measured by transwell chamber after exposure to 1.0 μg/ml TSP-1 and synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml) for different times (6, 12, 24, 48 hours). Caspase-3 and factor associated suicide (FAS) protein levels were measured by Western blot. The mRNA level of bcl-2 and FAS ligand (FASL) were measured by reverse transcription-polymerase chain reaction (RT-PCR). ResultsThe survival rate of RF/6A cells was determined by the treatment time and concentration of TSP-1(1.0 μg/ml) and the synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml). The lowest survival ratio of RF/6A was 78% (P < 0.001) when cells were treated by 10 μg/ml synthetic peptide VR-10 after 48 hours. TSP-1 and synthetic peptide VR-10 could inhibit migration of RF/6A cells in transwell chamber (P < 0.001). 10.0 μg/ml synthetic peptide VR-10 had the strongest effect, 1.0 μg/ml TSP-1 was the next. Migration inhibition rate was increase with the increase of the concentration of VR-10 (P < 0.001). There was no significant differences between 0.1 μg/ml and 1.0 μg/ml VR-10 (P=0.114). Western bolt showed that RF/6A cell in control group mainly expressed the 32×103 procaspase-3 forms. To 10.0 μg/ml VR-10 treated group, it showed decreased expression of procaspase-3 (32×103) and concomitant increased expression of its shorter proapoptotic forms (20×103). Compared with control group, expression of FAS peptides were significantly increased in 10.0 μg/ml VR-10 treated group. Compared with control group, expression of FasL mRNA was significantly increased in 10.0 μg/ml VR-10 treated group(t=39.365, P=0.001), but the expression of bcl-2 mRNA was decreased(t=-67.419, P=0.000). ConclusionTSP-1 and synthetic peptide VR-10 had the ability to inhibit proliferation and migration of endothelial cell, and also induce apoptosis by increasing FAS/FASL expression and repressing bcl-2 expression.
Objective To observe the effects on rabbit corneas and retinas after single intravitreal injection of voriconazole at different doses. Methods According to the randomization table, 25 healthy rabbits were randomly divided into control group, and voriconazole 50, 100, 200, and 400 μg groups. Therefore, there were 5 rabbits in each group. The eyes of control group received intravitreal injection of 0.1 ml balanced saline solution, and those treatment groups received 0.1 ml voriconazole injection of corresponding dose. Before the injection and 1, 7, and 14 days after the injection, endothelial cell counts and corneal thicknesses were measured; full-field electroretinogram were performed and b-wave amplitudes in maximal combined reaction (Max-R) were recorded. On 14 days after the injection, histologic structures were observed by light microscope and transmission electron microscope. Results There was no significant difference in endothelial cell counts (F=0.320, 0.291, 0.467, 0.649) and corneal thicknesses (F=0.214, 0.284, 0.360, 0.225) with those of control group at any time points (P > 0.05). Before and 1 day after the injection, b-wave amplitudes of each voriconazole group had no significant difference compared with those of control group (F=0.220, 0.106; P > 0.05). On 7 days after the injection, b-wave amplitudes decreased significantly at doses of 200 μg and 400 μg (P < 0.05). On 14 days after the injection, there was no significant difference between the the amplitude of 200 μg group and that of control group (P > 0.05). However, the amplitude of the 400 μg group decreased continuously and there was still significant difference (P < 0.05). Light microscopy did not reveal any corneal abnormality in both control group and voriconazole groups. The retinas were normal except that of the 400 μg group, which hadathinner and degenerated inner nuclear layer and disordered photoreceptor layer. Under transmission electron microscope, there were no ultrastructure damages of corneas in both control group and voriconazole groups, either. The rabbit retinas of the 50 μg and 200 μg group have normal inner nuclear layer and photoreceptor layer, but degrees of changes in both layers were observed in the eyes of 200 μg and 400 μg group. Conclusions There is no obvious effects on rabbit corneas and retinas after single intravitreal injection of voriconazole at he dose less than or equal 100 μg. There are no obvious effects on rabbit corneas at the dose of 200 μg and 400 μg, while there are damages to the retinas in both functions and histological structures.
Objective To observe the influence of the indomethacin on the proliferative and invasive activity of OCM-1 human choroidal melanoma cells. Methods OCM-1 cells were cultured with different concentrations of indomethacin (25, 50, 100, 200, 400 mu;mol/L ), and their proliferation were assessed by methyl thiazolyl tetrazolium（MTT）, invasive behaviors were examined by cell invasion assays, expression of survivin and VEGF were evaluated by reverse transcriptase polymerase chain reaction（RT-PCR）, immunofluorescence staining, ELISA and western blot analysis. Result All concentrations of indomethacin in this study can inhibit the proliferation and invasion of OCM-1 cells in a time and dosage-dependant manner（MTT/24 h：F=19.642，P＜0.01；MTT/48 h：F=136.597，P＜0.01；MTT/72 h：F=582.543，P＜0.01；invasion assays：F=54.225，P＜0.01）. Immunofluorescence staining indicated that survivin and VEGF mainly expressed in the cytoplasm of OCM-1 cells. Survivin mRNA in OCM-1 cells was inhibited by 100, 200, 400 mu;mol/L indomethacin（F=16.679，P＜0.01）. The concentrations of survivin were (787.3plusmn;47.37), (257.0plusmn;26.21), (123.3plusmn;8.02) pg/ml in control group and 100, 400 mu;mol/L indomethacin groups, respectively. Survivin expression was also significantly down-regulated in indomethacin-treated cells by Western blot analysis.Indomethacin had no effects on VEGF expression in OCM-1 cells.Conclusions Indomethacin can inhibit proliferation and invasion of OCM-1 cells in vitro,down-regulated expression of survivin may be the mechanism.
ObjectiveTo investigate the inhibitory effect of lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR).MethodsOne hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group, simple OIR model group, OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group), with 16, 32, 32 and 32 mice, respectively. When the mice were 7 days old, the mice in the normal control group were fed in a routine environment, and the mice in the OIR model group, Vec group and PSF group were established OIR model. The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1×1011 TU/ml) at the age of 12 days. No injection was performed in the normal control group and simple OIR group. RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Western blot analysis was applied to detect the protein expression of Nrf2, HO-1 and PSF. Results Of the normal control group, simple OIR model group, Vec group and PSF group, the number of pre-retinal neovascular cell nuclei were 0.00, 14.36±5.50, 15.67±4.96, 8.13±2.09, the non-perfusion area were 0.00%, (35.71±2.81)%, (36.57±4.53)%, (15.33±4.75)%, respectively. The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87, 165.70; P＜0.05). Compared with the normal control group, there were more pre-retinal neovascular cell nucleis and larger non-perfusion area in the simple OIR model group and Vec group (P＜0.05). Compared with the simple OIR model group and Vec group, there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P＜0.05). Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2, HO-1 (F=53.66, 83.54) and protein expression of Nrf2, HO-1 and PSF (F=58.38, 52.69, 24.79) among 4 groups were significant (P＜0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P＜0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P＜0.05). model group and Vec group (P＜0.05).ConclusionIntravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1.
Objective To observe the expression of alpha;A-and alpha;B-in retina after blue-light exposure.Methods Forty female Wistar rats were divided randomly into 4 groups:control group,and blue-light exposure for 6,12,and 24 hours groups, with 10 rats in each group. The rats in the control group were not intervened.The other three groups of rats were exposed to blue fluorescent lights for 6,12,and 24 hours respcetively. Then the rats were kept in darkness for 12 hours. The globes were enucleated after anaesthesia.The immunohistochemistry and Western blot were performed to detect the expression of alpha;A and alpha;B-crystallin in retina.Results The absorbance value (A value) of retina alpha;A-crystallin was 1.40573plusmn;0.70748 in the control group, and were 4.317 51plusmn;0.412 97, 7.397 08plusmn;1.947 90, 9.634 32plusmn;2.377 61, respectively in the other 3 groups; the difference among the groups was significant (F=24.569，P＜0.001). The A value of retina alpha;B-crystallin is 0.129 36plusmn;0.033 93 in the control group, and were 0.507 17plusmn;0.117 55, 7.345 43plusmn;2.292 97, 4.042 26plusmn;3.890 23, respectively in the other 3 groups; the difference among the groups was significant（F=40.102，P＜0.001）. The results of Western blot showed that the expression of alpha;A and alpha;B crystallin in groups with bluelight exposure was obviously higher than that in the control group.Conclusions Blue light may up-regulate the expression of alpha;A-and alpha;B-crystallin in ratsprime; retina.