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Objective:To observe the changes in blood flow density of radial retinal peripapillary capillary (RPC) around the optic disc in patients with non-arteritic anterior ischemic optic neuropathy (NAION) at different stages of the continuous course of the disease.Methods:A prospective cohort study. From January to December 2020, 29 cases of 29 eyes of NAION patients diagnosed in the Eye Center of the Second Affiliated Hospital of Zhejiang University School of Medicine were included in the study. Among them, there were 18 males with 18 eyes and 11 females with 11 eyes. The average age was 53.62±6.67 years old. The affected eye underwent routine eye examination and visual field, optic cohenrence tomography angiography (OCTA) examination. Visual field inspection was performed to obtain the average visual mean defect (MD) value. OCTA was used to measure the thickness of the peripapillary retinal nerve flayer (pRNFL) around the optic disc, the whole en face image vessel density (wiVD), intro disc vessel density (diVD), RPC blood flow density around the optic disc, and macular ganglion cell complex (GCC). The course of disease ≤3 weeks was defined as the acute phase; 4-12 weeks was defined as the subacute phase; >12 weeks was defined as the chronic phase. The changes of visual field MD, optic disc RPC blood flow density, pRNFL thickness and macular GCC thickness were observed in the acute, subacute and chronic phases (12-24, >24 weeks). A completely randomized design of variance analysis was used to compare the differences in visual field MD, RPC blood flow density, GCC, and pRNFL thickness in different courses. Pearson correlation analysis was used to analyze the correlation between pRNFL thickness, macular GCC thickness, visual field MD changes and RPC blood flow density around the optic disc sex.Results:The wiVD of the eyes in the acute phase, subacute phase, and chronic phase (12-24 weeks, >24 weeks) were (44.96±2.76)%, (41.50±3.49)%, (39.08±5.43)%, (38.56±6.48)%. There was a statistically significant difference in wiVD of eyes with different disease courses ( F=8.939, P<0.001). The average difference of wiVD between 12-24 weeks and >24 weeks in the chronic phase was -0.984, and the difference was not statistically significant ( P>0.05). There was no statistically significant difference in diVD of patients with different courses of disease ( F=1.079, P=0.365). The blood flow density of RPC around the optic disc of the affected eye, except for the lower part, the blood flow density of the nasal side, the temporal side, and the upper quadrant, decreased significantly with the progression of the disease, and the difference was statistically significant ( F=8.816, 6.069, 8.943; P<0.05). In the chronic phase, the average difference of blood flow density between the nasal, temporal, and upper sides of the eyes between 12-24 weeks and more than 24 weeks in the chronic phase was -0.984, -0.230, -0.198, and the difference was not statistically significant ( P>0.05). There was no statistically significant difference in the visual field MD of patients with different courses of disease ( F=0.277, P=0.842); the overall pRNFL thickness and average macular GCC thickness were compared with statistical significance ( F=47.122, 14.954; P<0.001, <0.001), all became significantly thinner with the progression of the disease. The results of Pearson correlation analysis showed that the blood flow density of the entire optic disc wiVD, the blood flow density of RPC in the temporal quadrant around the optic disc and the visual field MD ( r=-0.225, -0.268; P<0.05), and the average thickness of GCC ( r=0.480, 0.436; P<0.01) were all related. Conclusion:The blood flow density of RPC in the entire optic disc and around the optic disc (except the lower quadrant) of NAION eyes gradually decrease with the progression of the disease, and stabilize after 12 weeks of the disease.
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Objective To investigate the effects of lentivirus-mediated knockdown of astrocyte elevated gene-1 (AEG-1) on the biological behavior of uveal melanoma(UM).Methods MUM-2B and MUM-2C cell lines with different invasiveness and metastasis potential were cultured.Based on AEG-1 sequence that designed the effective (RNA interference) RNAi target sequence and the RNAi negative control scramble sequence,and then performed the preparation of the lentiviral vector and viral packaging.In these two kinds of cell lines,the cells infected by AEG-1-RNAi lentivirus were used as lentiviral infection group,the cells of the RNAi negative control of the blank lentivirus infected cells were used as negative control group,and the cells without any processing were used as normal control group.Real-time PCR and Western blot were used to detect the change of AEG-1 transcription and protein expression levels in these two kinds of cell lines' groups,as well as by MTT method,Annexin V-APC method,cell invasion assay,and Transwell cell migration assay that to investigate the effect of lentivirus transfection induced knockdown of AEG-1 on the biological behavior of human UM cell lines.Results The successful preparation of RNAi lentivirus vector and its viral package were transfected to MUM-2B,MUM-2C cell lines of logarithmic growth phase,respectively.The relative expressions of AEG-1 mRNA in normal control groups,negative control groups and lentiviral infection groups were statistically significant (F =130.02,P＜0.01;F =144.17,P＜0.01).The relative expression of AEG-1 mRNA in lentiviral infection groups were lower than that in the negative control groups,and the AEG-1 gene knockout rates were 77.1％ and 79.8％,respectively,the differences were statistically significant (both at P＜0.01).There were no significant differences in the relative expression of AEG-1 mRNA between normal control groups and negative control groups (all at P＞0.05).Western blot showed that no significant changes were found in the AEG-1 proteins expression levels between normal control groups and negative control groups in the two kinds of cell lines (F=146.17,P＜0.01;F=156.79,P＜0.01).The expression levels of AEG-1 protein in the lentiviral infection groups were significantly lower than that in negative control groups (all at P＜0.01).The proliferation of both cells lines in lentiviral infection groups were significantly lower than that in negative control groups from the second day to the fifth day,the differences were statistically significant (t =5.78,30.68,23.99,29.40,all at P ＜ 0.01;t =7.88,7.09,5.56,6.60,all at P ＜ 0.01).In both cell lines,the apoptosis rate of lentiviral infection groups were significantly higher than those in the negative control groups,the differences were statistically significant (t =54.34,P＜0.01;t =11.68,P＜0.01).In both cell lines,the multiple of invasions in lentiviral infection groups were significantly lower than those in negative control groups,and the differences were statistically significant (t =16.04,P＜0.01;t =13.98,P＜0.01);In both cell lines,the multiple of transfer in lentiviral infection groups were significantly lower than those in negative control groups,and the differences were statistically significant (t =12.04,P ＜ 0.01;t =22.43,P ＜ 0.01).Conclusions Lentivirus transfection inducing knockdown of AEG-1 inhibits the transcription and protein expression level of AEG-1 in the melanoma cells,which changes the biological behaviors of UM,like slowing down cell proliferation,promoting apoptosis,and reducing the abilities of cells' invasion and metastasis.
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Background Drug resistance is the main cause of failure after chemotherapy of retinoblastoma (RB),and how to improve the sensitivity of RB cells to chemotherapic drug become an urgent issue.All trans retinoic acid (ATRA) can inhibit the growth of tumor cells.However,whether ATRA increases the sensitivity of RB cells to chemotherapic drug is unclear.Objective This study aimed to investigate the inhibitory effect of ATRA with vincristine on the proliferation of SO-RB50 cells.Methods SO-RB50 cells were cultured routinely.Different concentrations of ATRA or vincristine were added into the medium for 48 hours for the determination of IC50 by cell counting kit-8 (CCK-8) method.Cultured cells were divided into normal control group,ATRA group,vincristine group and combined drug group.The cells were treated by ATRA or vincristine with the dose of IC50,and the proliferation of the cells was detected every day at the 24-hour interval for 6 consecutive days.The percentage of the cells in different cell cycles was analyzed 72 hours after treatment using flow cytometry.Cell apoptosis rate was detected and calculated 48 hours after treatment by annexin V/propidium iodide (PI) method.Results The IC50 of ATRA was approximately 12.84 μ mol/L,and that of vincristine was 0.11 μg/ml.The growth curve of SO-RB50 cells was gradually raised as the lapse of time,but the curves were relatively low in the ATRA group,vincristine group and combined drug group,with the lowest curve in the combined drug group.The proliferation values of the cells (A450)were 1.078±0.022,0.611 ±0.038,0.596 ±0.031 and 0.483 ±0.030 in 48 hours after treatment,and those in 72 hours were 1.380± 0.021,0.799 ± 0.016,0.668 ± 0.041 and 0.532 ± 0.033 in normal control group,ATRA group,vincristine group and combined drug group,showing significant differences among the groups and various time points (Fgroup =1115.207,P =0.000;Ftime =257.781,P =0.000).The A450 values of the ATRA group,vincristine group and combined drug group were significantly lower than those of normal control group (all at P＜0.05).The percentage of the cells in different cell cycles was changed after 72 hours' treatment and the differences were statistically significant (FG0/G1 =130.565,Fs =57.435;FG2/M =114.290,P＜0.05).Compared with the normal control group,the percentage of G0/G1 phase cells was increased and S phase cells was decreased significantly in ATRA group,the percentage of G0/G1 phase cells was decreased and G2/M phase cells was increased significantly in vincristine group(P＜0.05).The apoptotic rate of the cells was (7.17±0.18) ％,(27.34±1.36) ％,(27.49±2.45) ％ and (34.50±1.84) ％ in normal control group,ATRA group,vincristine group and combined drug group,with a significant difference among the groups (F=147.555,P＜0.05),and the apoptotic rate in the combined drug group was remarkedly lower than that of normal control group,ATRA group and vincristine group (all at P=0.000).Conclusions ATRA can improve the sensitivity of SO-RB50 cells to chemotherapeutic drug.The combined application of ATRA and vincristine enhance the inhibitory effect on the cells probably by regulating cell cycle and inducing apoptosis.
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Objective To analyse characteristics of military pilot group sandplay work and reveal their psychological status.Methods Using the self-reporting 90-item Symptom Checklist (SCL-90) to test the validity of group sandplay work made by 87 groups of military pilots.Statistical analysis was used to analyse the characteristics of code data in group sandplay work.Results Positive and negative themes in group sandplay work showed negative and positive correlations(r=-0.59,0.59) with symptom factors of SCL-90,respectively.Positive and negative themes scores were 5.37±2.25 and 2.36± 1.85 ,respectively.The secondary theme scores in the positive theme were entirely higher than those in the negative theme.Scores of energy, connection, cooperation, integration, relaxation, and spirituality in the positive theme and threat,limitation, and aggression in the negative theme were relatively high.The overall evaluations of the work include power, integration, enrichment, and fluency.The theme names of the work include life, military, natural and abstract.Conclusion Group sandplay is an effective method in the study of military pilots' psychological health and military pilots are in good mental health.These pilots have strong intrinsic energy,teamwork and communication.However,some pilots have symptoms of anxiety,tension and insecure.