Objective To observe the expression of p53, bcl-2 genes, vascular endothelial cell growth factor（VEGF）, basic fibroblast growth factor（bFGF）, insulin-like growth factor-I （IGF-I）, and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1- to 20-week diabetic rats, and the relationship between the expressions and cell cycle arrest.Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl-2. Results Under LM, immunohistochemical positive expression of p53 and bcl-2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8- to 20-week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF-I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20-week diabetic rat expressed p53 and bcl-2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl-2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1- to 20-week diabetic rats. Conclusion p53, bcl-2 may introduce cell cycle arrest of VECs of retinae in 8- to 20-week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF-I and their receptors, and the growth factors may keep VECs surviving by self-secretion. (Chin J Ocul Fundus Dis，2003，19：29-33)
ObjectiveTo observe the expression of Rap1, guanosine triphosphate-Rap1 (GTP-Rap1), vascular endothelial growth factor (VEGF) and β-catenin in experimental choroidal neovascularization (CNV).MethodsForty-two brown Norwegian rats were randomly divided into a blank control group (7 rats) and a model group (35 rats). Both eyes were enrolled. The CNV model was established by holmium ion laser photocoagulation in the model group. At 3, 7, 14, 21, and 28 days after photocoagulation, fluorescein fundus angiography (FFA) and choroidal vascular smear were performed to observe the degree of fluorescein leakage and CNV area in rats; Western blot and real-time quantitative polymerase chain reaction (RT-PCR) were used to detect the expression of Rap1, GTP-Rap1, VEGF, β-catenin and mRNA in CNV.ResultsThe results of FFA examination showed that a large disc-shaped fluorescein leaked in the photo-condensation spot 14 days after photocoagulation. Laser confocal microscopy showed that compared with 7 days after photocoagulation, CNV area increased at 14, 21, 28 days after photocoagulation, and the difference were statistically significant (t=3.725, 5.532, 3.605;P＜0.05). Western blot showed that there was no significant difference in the relative expression of Rap1 protein in CNV at different time points after photocoagulation between the two groups (P=0.156). Compared with the blank control group, the relative expression of GTP-Rap1 protein was significantly decreased, the relative expression of VEGF and β-catenin protein were significantly increased in the model group (P=0.000). The results of RT-PCR showed that there was no significant difference in the relative expression of Rap1 mRNA at different time points after photocoagulation between the two groups (P=0.645), but there were significant difference in the relative expression of β-catenin mRNA (P=0.000). At 7, 14, 21 and 28 days after photocoagulation, there were significant difference in the relative expression of GTP-Rap1 and VEGF mRNA between the two groups (P=0.000).ConclusionsThe expression of GTP-Rap1 in experimental CNV is significantly lower than that in normal rats.
Objective To investigate the expression of pigment epitheliumderived factor (PEDF) and vascular endothelial growth factor (VEGF) in choroidal melanoma. Methods The expression of VEGF and PEDF protein in fifty-eight cases of paraffinembeded choroidal melanoma samples was measured by immunohistochemistry, the expression of PEDF mRNA in thirtynine choroidal melanoma samples was assayed by in situ hybridization. Results PEDF protein was detected in 13/58 cases (22.4%) of choroidal melanoma, the positive rate in nonsclerainvasion group (12/38, 31.6%) was higher (Plt;0.05) than that in sclerainvasion group (1/20, 5%). VEGF protein was detected in 43/58 cases (64%) of choroidal melanoma, the positive rate in nonsclerainvasion group (25/38, 65.8%) was lower (Plt;0.05) than that in sclerainvasion group (18/20, 90%). The expression of PEDF mRNA was detected in 18/39(46.2) cases, the positive rate in nonsclerainvasion group was higher (Plt;0.05) than that in sclerainvasion group. Conclusions Imbalanced expression of VEGF and PEDF in choroidal melanoma may play a key role in the angiogenesis, tumor progression and metastasis.
Objective To investigate the inhibitive effect of E2F decoy oligodeoxynucleotides (E2F decoy ODNs) on cultured human retinal pigment epithelial (HRPE) cells.Methods E2F decoy ODNs or scramble decoy ODNs at varied concentrations were put into the HRPE cells mediated by lipofectamineTM2000. The proliferative activity of HRPE was detected by methythiazolyl-terazollium assay, and the competitive combinative activity of E2F decoy ODNs and transcription factor E2F was detected by electrophoresis mobility-shift assay. Results The proliferation of HRPE was inhibited markedly by E2F decoy ODNs at the concentration of 0.2 μmol/L (P=0.002) in a dose-dependent manner but not by scrambled decoy. The results of electrophoresis mobility-shift assay showed that the combinative activity of transcription factor E2F was abolished completely by E2F decoy ODNs. Conclusions E2F decoy ODNs may sequence-specifically inhibit the combinative activity of transcripti on factor E2F,and inhibit the proliferation of HRPE cells.(Chin J Ocul Fundus Dis，2004，20：182-185)
Objective To observe the incidence of hypotony and its risk factors in the early stage after 23-gauge sutureless microincisional vitrectomy. Methods A retrospective case series. Seventy patients (70 eyes) who had undergone 23-gauge sutureless microincisional vitrectomy were enrolled. There were 41 phakic eyes, 29 eyes with intraocular lens; 68 eyes with a single surgical procedure and 2 eyes with a second surgical procedure; 34 eyes with BSS tamponade and 36 with gas tamponade. Intraocular pressure (IOP) was measured by non-contact tonometry. Hypotony was defined as an IOP of 5 mm Hg (1 mm Hg=0.133 kPa ) or less. The incidence of hypotony and other complications at postoperative day 1, 3 and 7 were observed. The influence of age, sex, side of operation, type of tamponade, status of lens, surgical time on postoperative hypotony was analyzed. Results Hypotony was found in 8 eyes (11.4%) on postoperative day 1 and recovered spontaneously on postoperative day 3. There were no significant differences comparing age (t=1.12), sex (chi;2=2.23) and side of operation (chi;2=2.01) between patients with hypotony and those without it. The patients with hypotony suffered longer surgical time than that of those without hypotony (chi;2=5.48,P＜0.05). The incidence of hypotony in eyes with gas tamponade was significantly lower than that in eyes with BSS tamponade (chi;2=5.48,P＜0.05). The incidence of hypotony in eyes with phakic eyes was lower than that in eyes with intraocular lens (chi;2=4.20,P＜0.05). Hypotony was encountered in the 2 re-operated eyes. Choriodal folds were encountered in 2 eyes, but there was no other complication in other eyes. Conclusions A transient hypotony occurs commonly in first 3 days after 23-gauge sutureless microincisional vitrectomy. Hypotony was significantly influenced by type of tamponade, reoperation and intraoperative lens status.
ObjectiveTo investigate the clinical characteristics and contribution factors in severe coronavirus disease 2019 (COVID-19).MethodsThe clinical symptoms, laboratory findings, radiologic data, treatment strategies, and outcomes of 110 COVID-19 patients were retrospectively analyzed in these hospitals from Jan 20, 2020 to Feb 28, 2020. All patients were confirmed by fluorescence reverse transcription polymerase chain reaction. They were classified into a non-severe group and a severe group based on their symptoms, laboratory and radiologic findings. All patients were given antivirus, oxygen therapy, and support treatments. The severe patients received high-flow oxygen therapy, non-invasive mechanical ventilation, invasive mechanical ventilation or extracorporeal membrane oxygenation. The outcomes of patients were followed up until March 15, 2020. Contribution factors of severe patients were summarized from these clinical data.ResultsThe median age was 50 years old, including 66 males (60.0%) and 44 females (40.0%). Among them, 45 cases (40.9%) had underlying diseases, and 108 cases (98.2%) had different degrees of fever. The common clinical manifestations were cough (80.0%, 88/110), expectoration (33.6%, 37/110), fatigue (50.0%, 55/110), and chest tightness (41.8%, 46/110). Based on classification criteria, 78 (70.9%) non-severe patients and 32 (29.1%) severe patients were identified. Significant difference of the following parameters was found between two groups (P<0.05): age was 47 (45, 50) years vs. 55 (50, 59) years (Z=–2.493); proportion of patients with underlying diseases was 27 (34.6%) vs. 18 (56.3%) (χ2=4.393); lymphocyte count was 1.2 (0.9, 1.5)×109/L vs. 0.6 (0.4, 0.7)×109/L (Z=–7.26); C reactive protein (CRP) was 16.2 (6.5, 24.0) mg/L vs. 45.3 (21.8, 69.4) mg/L (Z=–4.894); prothrombin time (PT) was 15 (12, 19) seconds vs. 18 (17, 19) seconds (Z=–2.532); D-dimer was 0.67 (0.51, 0.82) mg/L vs. 0.98 (0.80, 1.57) mg/L (Z=–5.06); erythrocyte sedimentation rate (ESR) was 38.0 (20.8, 59.3) mm/1 h vs. 75.5 (39.8, 96.8) mm/1 h (Z=–3.851); lactate dehydrogenase (LDH) was 218.0 (175.0, 252.3) U/L vs. 325.0 (276.5, 413.5) U/L (Z=–5.539); neutrophil count was 3.1 (2.1, 4.5)×109/L vs. 5.5 (3.7, 9.1)×109/L (Z=–4.077). Multivariable logistic analysis showed that there was positive correlation in elevated LDH, CRP, PT, and neutrophil count with the severity of the disease. Currently, 107 patients were discharged and 3 patients died. Total mortality was 2.7%.ConclusionsOld age, underlying diseases, low lymphocyte count, elevated CPR, high D-dimer and ESR are relevant to the severity of COVID-19. LDH, CPR, PT and neutrophil count are independent risk factors for the prognosis of COVID-19.
ObjectiveTo analyze the causes and potential risk factors of re-catheterization after failure of no urinary catheter in patients undergoing lung cancer surgery.MethodsThe clinical data of 1 618 patients without urinary catheter indwelling during the perioperative period of thoracic surgery in our hospital from 2013 to 2019 were retrospectively analyzed, including 791 males and 827 females, with a median age of 58 years, ranging from 27 to 85 years. And the risk factors for re-insertion after failure of urinary catheter were investgated.ResultsThe rate of catheter re-insertion was 1.5% (24/1 618). Compared with patients without re-insertion, patients with re-insertion had longer operation time [120.0 (95.0, 130.0) min vs. 120.0 (115.0, 180.0) min, P=0.015] and more intraoperative fluid infusion [800.0 (600.0, 1 100.0) mL vs. 1 150.0 (725.0, 1 350.0) mL, P=0.008]. Further multivariate analysis found that the operation time (OR=1.014, P=0.004, 95%CI 1.005-1.024) and intraoperative fluid infusion (OR=1.001, P=0.022, 95%CI 1.001–1.002) were independent risk factors for re-insertion.ConclusionThe rate of catheter re-insertion in lung cancer patients is relatively low, and conventional no placement of catheter is safe and feasible after lung cancer surgery. Increasing operation time or intraoperative infusion volume may increase the risk of catheter re-insertion after lung cancer surgery.
Objective To observe the therapeutic effect of vitrectomy on Eales disease and the correlated factors affecting the visual prognosis and disease outcomes. Methods The clinical and follow-up data from 128 patients (142 eyes) with Eales disease undergone vitrectomy were retrospec tively analyzed. The statistical methods including t test,chi;2test, one-way Anova method of square-deviation(SD), and logistic regression were used to analyze the relationship between the general data of the patients (including age, sex, laterality of the eye, visual acuity before the treatment, stages of disease, duration from vitreous hemorrhage to vitrectomy, neovascularization and proliferative vitreoretinopathy, and whether or not combined with retinal detachment and other complications) and the prognosis of the visual acuity after surgery (including the surgical method, techniques, and times and complications after the surgery). The patients were 18-45 years old (mean 28.5 years) with single vitreous hemorrhage in 28 and proliferative changes in 114 in whom 59 had retinal detachment. The follow-up period after the surgery was more than 3 months (mean 35.8 months). The success criteria of the surgery were complete or part retinal reattachment, and failure of retinal reattachment, eye-ball atrophy or excis ion of the affected eye were the failure criteria. Results Successful vitrectomies had been performed on 129 eyes (90.8%) and unsuccessful ones on 13 eyes (9.2%). The difference between the stages of the disease and prognosis of visual acuity after the surgery was significant (chi;2=64.0, Plt;0.01); the duration of vitreous hemorrhage obviously affected the prognosis of visual acuity (OR=11.6，Plt;0.01); the degree, quality, curable possibility, and recurrent probability of combined retinal detachment were the key factors affecting the visual acuity after vitrectomy; the visual acuity before and after the surgery was interrelated; the method and techniques of the surgery and the different filling matters affected the visual acuity after the surgery; the difference between multiple times and once of surgery was significant (chi;2=66.84，Plt;0.01); the degree of complexity of the operative procedure, especially repetitious vitrectomies obviously affected the surgical prognosis and the improvement of visual acuity; the possibility of fa ilure of the surgery differs 7 times in patients with or without post-operative complications ( chi;2=67.23，Plt;0.01); whether the post-operative complications occurred or not significantly affected the prognosis of the visual acuity a-fter the surgery. Conclusions Vitrectomy is effective for Eales disease. The important factors affecting the prognosis of visual acuity after the operation include stages of disease, degree and extent of proliferative vitreoretinopathy, whether or not combined with retinal detachment and other complic ations, duration from vitreous hemorrhage to vitrectomy, the degree of complexity of the operation, and the complications during or after the operation. (Chin J Ocul Fundus Dis, 2006, 22:291-294)
Objective To verify the effect of Evans blue dye on determining the retina blood vessel leakage． Methods Male Sprague-Dawley rats were used in this study． The VEGF induced retinal blood vessel leakage was checked with Evans blue dye． Then the bloodretina barrier breakdown of 1 week diabetic animals was quantified with Evans blue．The dye was extracted from retina by formamide and the extraction was checked with spect rophotometer． Evans blue leakage was normalized against wet or dry retina weight． Results The retinal Evans blue content of eyes treated with VEGF was remarkably higher than that of the controls (n=17 ,Plt;0．0001)． And the eyes of 1 week diabetic duration animals had more Evans blue dye than that of the normal controls (Plt;0．05)． Conclusion Evans blue dye is a sensitive tracer in quantitatively diagnosing the blood retina barrier breakdown． (Chin J Ocul Fundus Dis, 2001,17:221-223)
Objective To investigate if lactic acid can promote the expression of vascular endothelial growth factor (VEGF) in the rat retinal explants.Methods The retinas of two-week neonatal SD rats were placed onto the culture plate inserts and incubated with Dulbeccoprime;s modified Eagleprime;s medium （DMEM） plus 2% fetal bovine serum (FBS) containing 10,20,30 mmol/L of lactic acid, respectively. Each group had 24 retinas. At 24 hours after incubation, the retinas were sectioned for light microscopy and the expression of VEGF was measured by real time PCR and Western blot. Results The cultured retinas maintained intact construction, and no cytolysis and apoptosis were observed under light microscope. RT-PCR showed the levels of VEGF mRNA were 0.74plusmn;0.06 for 10 mmol/L lactic acid group, 0.99plusmn;0.12 for 20 mmol/L group, and 1.45plusmn;0.17 for 30 mmol/L group respectively. VEGF expression was 0.34plusmn;0.15 for 10 mmol/L, 0.54plusmn;0.16 for 20 mmol/L, and 0.93plusmn;0.23 for 30 mmol/L group respectively by Western blot. Both PCR and Western blot showed 30 mmol/L of lactic acid significantly increased the levels of VEGF mRNA and VEGF expression. Conclusion The induction of retinal VEGF by lactic acid is concentration-dependent.