OBJECTIVE:To observe the effect of dexamethasone to intracellular free Ca2+ of frozen RPE cells. METHODS:The cultured human RPE cells were frozen for 30s at --70deg;C. The RPE cells were loaded with Fura-2/AM and analyzed using a digital imaging microscopy system,the effect of dexamethasone to intracellular free Ca2+ was measured at a serial concentration of 40, 60,100,150,200mu;g/ml. RESULTS:The concentration of intracellular free Ca in frozen human RPE cells was increased to 18.6%~29.8% by dexamethasone at concenlration of 40mu;g/ml~60mu;g/ml,while was decreased to 28.4%~35.2% at 150mu;g/ml~200mu;g/ml. CONCLUSIONS:Effect of dexamethasone showed two aspects of effect to frozen cultured human RPE ceils,that it was inhibitor at high concentration and stimulator at low concentration （Chin J Ocul Fundus Dis,1997,13: 86-88）
Objective To evaluate the efficacy and safety of dexamethasone intravitreal implant (Ozurdex) in the treatment of macular edema (ME) secondary to retinal vein occlusion (RVO). Methods Thirty-nine patients (39 eyes) with ME secondary to RVO were enrolles in this study. Of the patients, 27 were male and 12 were female. The mean age was (41.9±16.3) years. The mean course of disease was (5.0±5.3) months. The best corrected visual acuity (BCVA), intraocular pressure and optical coherence tomography (OCT) were performed. BCVA was measured by Early Treatment Diabetic Retinopathy Study charts. Central macular thickness (CMT) was measured by OCT. The mean BCVA was (13.4±15.3) letters. The mean intraocular pressure (IOP) was (14.1±2.8) mmHg (1 mmHg=0.133 kPa). The mean CMT was (876.1±437.9) μm. Of the 39 eyes, 33 were central RVO, 6 were branch RVO. Patients were categorized into ischemic (18 eyes)/non-ischemic (21 eyes) groups and previous treatment (22 eyes)/treatment naïve (17 eyes) groups. All eyes underwent intravitreal 0.7 mg Ozurdex injections. BCVA, IOP and CMT were assessed at 1, 2, 3, 6, 9, 12 months after injection. Three months after injection, intravitreal injections of Ozurdex, triamcinolone acetonide or ranibizumab could be considered for patients with ME recurrence or poor treatment effects. Change of BCVA, IOP and CMT were evaluated with paired t test. The presence of ocular and systemic adverse events were assessed. Results BCVA, IOP significantly increased and CMT significantly decreased at 1 month after injection compared to baseline in all groups (t=3.70, 3.69, 4.32, 3.08, 4.25, 6.09, 6.25, 4.02, 5.49, 8.18, 6.54, 5.73; P＜0.05). Two months after injection, change of BCVA, IOP and CMT was most significant (t=4.93, 6.80, 6.71, 5.53, 4.97, 5.89, 5.13, 7.68, 7.31, 8.67, 8.31, 5.82; P＜0.05). Twelve months after injection, there was no statistical difference regarding BCVA of ischemic RVO group and previous treatment group, compared to baseline (t=1.86, 0.67; P＞0.05); BCVA of non-ischemic RVO group and treatment naïve group significantly increased compared to baseline (t=2.27, 2.30; P＜0.05); there was no statistical difference regarding IOP in all groups (t=0.30, 0.13, 0.64, 1.53; P＞0.05);however, CMT significantly decreased in all groups (t=4.60, 3.26, 3.00, 4.87; P＜0.05). Twenty-seven eyes (69.2%) experiences ME recurrence (4.5±1.5) months after injection. Most common side-effect was secondary glaucoma. 41.0% eyes had IOP more than 25 mmHg, most of which were lowered to normal range with use of topical IOP lowering drugs. Four eyes (10.3%) presented with significant cataract progression and needed surgical treatment, all were central RVO eyes. No serious ocular or systemic adverse events such as vitreous hemorrhage, retinal detachment or endophthalmitis were noted. Conclusions Intravitreal injection of Ozurdex for patients with ME secondary to RVO is effective in increasing BCVA and lowering CMT in the first few months. Significant treatment effect could be seen at 1 month after injection and was most significant at 2 months after injection. The long-term vision of eyes in non-ischemic RVO group and treatment naïve group are better. 69.2% eyes experience ME recurrence at 4 months after injection. Short term adverse events were mostly secondary glaucoma and long term adverse events are mostly cataract progression.
Objective To investigate the therapeutic effect of dexamethasone on children with severe community acquired pneumonia ( CAP) . Methods 120 children with severe CAP admitted from January 2009 to June 2011 were recruited in the study. The patients were randomly divided into a dexamethasone group ( n = 62) and a control group ( n = 58) . The patients in the dexamethasone group received additional dexamethasone intravenous injection for 3 days ( 0. 2-0. 4 mg· kg- 1 · d- 1 , qd) on the basic treatment of the control group. Length of hospital stay, serum C reactive protein ( CRP) concentration on 4th day after admission, overall efficacy, mortality, incidence of adverse events during treatment were compared between the two groups. Results Median length hospital stay was 8 days in the dexamethasone group compared with 9 days in the control group without significant difference ( P gt;0. 05) . The serumCRP concentration on 4th day was lower in the dexamethasone group than that in the control group [ ( 23. 4 ±5. 6) mmol /L vs. ( 41. 3 ±6. 2) mmol /L, P lt;0. 05] . The overall efficacy was higher in the dexamethasone group than that in the control group ( 88. 7% vs. 74. 1% , P lt; 0. 05) . The in-hospital mortality and incidence of severe adverse events were not significantly different between the two groups ( P gt; 0. 05) . Conclusions Dexamethasone treatment is associated with a significant attenuation in systematic inflammatory response, but does not decrease mortality in hospitalized children with severe CAP.
Objective To investigate the mechanism of dexamethasone in the treatment of acute necrotizing pancreatitis (ANP). Methods The ANP of 48 SD rats were induced by retrograde infusion of sodium taurocholate through biliopancreatic duct.After 30 minutes,the therapy group was administrated with dexamethasone at a dose of 0.2 mg/100 g alone. The control group was administrated with the same amount of 0.9% saline solution.At fourth hour and twelfth hour,8 rats of each group were sacrificed to examine the levels of serum tumor necrosis factor-alpha(TNFα) and serum amylase,to score the degree of pancreatic necrosis and to evaluate acinar cell apoptosis by in situ hybridization by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL). The survial period of 8 rats in each group were observed. Results In therapy group, the level of TNFα was (17.8±2.7) pg/ml and (8.5±1.6) pg/ml,the apoptosis index was （36.94±4.12）% and （ 32.79±3.31）%,the survival period was (33.4±21.5) h.While the control group with the indexes mentioned above were as follows: (53.6±18.7) pg/ml and (37.2±11.1) pg/ml ( P＜0.01),（4.37±1.24）% and （5.12±2.11）% （P＜0.01）,(14.6±5.7) h (P＜0.01) ,the histologic scoring for ANP between therapy group and control group was a significantly distinct (P＜0.01). Conclusion Dexamethasone can induce pancreatic acinar cell apoptosis in this model. Proper leves of TNFα may play an important role in regulating the apoptosis.Apoptosis can protect pancreas from necrosis in ANP.
ObjectiveTo observe the long-term effects of anti-vascular endothelial growth factor (VEGF) drug initiation combined with dexamethasone intravitreal implant (DEX) on the structural integrity of the outer macular region of the eye in patients with macular edema (ME) secondary to central retinal vein occlusion (CRVO). MethodsA retrospective clinical study. From February 2018 to August 2022, 54 patients diagnosed with CRVO combined with ME (CRVO-ME) in Department of Ophthalmology of Central Theater Command General Hospital were included in the study. Among them, there were 30 males and 24 females, all with monocular disease. According to different treatment regiments, patients were divided into anti-VEGF and DEX combination therapy group (initial combination group), anti-VEGF drug monotherapy group (monotherapy group) with 21 eyes and 33 eyes, respectively. Best corrected visual acuity (BCVA), optical coherence tomography (OCT) examination were performed in all eyes. The thickness of foveal retina (CRT) and the deficiency length of outer membrane (ELM), ellipsoid band (EZ) and chimaera band (IZ) in the 1 mm macular area were measured by OCT. The initiating combination group was treated with anti-VEGF agents or DEX as assessed on demand (PRN) after the combination therapy, and the monotherapy group received 3+PRN regimen. Relevant examinations were performed at 1 (V1), 6 (V6), 12 (V12) months and observation cut-off or the last visit (Vf) after treatment using the same equipment before treatment. The deletion length of ELM, EZ and IZ in V1, V6, V12 and Vf after treatment were compared between the two groups. Repeated measurement ANOVA was used to compare BCVA, CRT and deletion length of ELM, EZ and IZ at different follow-up times. Spearman rank correlation test was used to analyze the correlation between the two groups of continuous variables. ResultsThe follow-up time of patients in the initial combination group and monotherapy group was (18.05±5.66) and (21.90±10.80) months, respectively, with no statistical significance (F=13.430, P=0.229). Compared with baseline, the deletion lengths of ELM, EZ and IZ were significantly improved (F=11.848, 10.880, 29.236), BCVA was increased (F=10.541) and CRT was decreased (F=52.278) in the initial combination group and the monotherapy group at different follow-up times after treatment. The differences were statistically significant (P＜0.001). At V1, EZ and IZ deletion lengths were (344.10±413.03), (593.33±372.96) μm and (354.71±321.75), (604.85±385.77) μm in the initial combination group and monotherapy group, respectively. The improvement of EZ and IZ deletion lengths in the initial combination group was better than that in the single drug group, and the difference was statistically significant (F=5.272, 6.106; P=0.026, 0.017). The CRT of the initial combination group and the monotherapy group were (248.86±59.99) and (314.72±214.91) μm, respectively, and the CRT of the initial combination group was significantly lower than that of the monotherapy group, with statistical significance (F=6.102, P=0.017). At V6, V12 and Vf, the deletion length of ELM, EZ and IZ and BCVA and CRT showed no statistical significance (P＞0.05). Correlation analysis showed that ELM, EZ, IZ were positively correlated with BCVA and CRT in the initial combination group and monotherapy group (P＜0.001). In V6, V12 and Vf, the number of anti-VEGF drug injections in the initial combination group and monotherapy group was (2.67±1.32), (4.43±2.27), (6.05±3.51), (4.58±0.90), (7.33±1.93), (11.33±6.10) times, respectively. The number of injections in the initial combination group was significantly lower than that in the monotherapy group, and the difference was statistically significant (F=5.150, 0.646, 3.433; P＜0.001). ConclusionsThe improvement of BCVA and CRT in the initial combination group is similar to that in the monotherapy group. Compared with the monotherapy group, EZ and IZ deletion are improved more significantly in the initial combination group, and CRT decreased more rapidly and significantly. The initial combination group receives fewer anti-VEGF injections than the monocular group.
ObjectiveTo observe the short-term efficacy and safety of a new strategy of dexamethasone intravitreal implant (DEX) combined with ranibizumab in the treatment of retinal vein occlusion (RVO) secondary to macular edema (ME) (RVO-ME). MethodsA prospective clinical interventional study. From May 2020 to September 2021, 78 RVO-ME patients with 78 eyes diagnosed in the eye examination of Department of Ophthalmology of The First Affiliated Hospital of Anhui University of Science&Technology were included in the study. Among them, there were 35 males and 43 females, all with monocular disease. Branch retinal vein occlusion (BRVO) was found in 40 patients with 40 eyes; central retinal vein occlusion (CRVO) was found in 38 patients with 38 eyes. According to the treatment strategies, patients were randomly divided into DEX and ranibizumab combination therapy group (initial combination therapy group), DEX monotherapy group and ranibizumab monotherapy group, with 29 eyes, 26 eyes and 23 eyes respectively. Different types of RVO were divided into different treatment groups of BRVO and CRVO. Best corrected visual acuity (BCVA) and frequency domain optical coherence tomography were performed. The BCVA examination was carried out using the international standard visual acuity chart, which was converted into the logarithmic minimum angle of resolution (logMAR) visual acuity during statistics. There were no significant differences in logMAR BCVA (χ2=2.376) and central retinal thickness (CRT) (F=0.052) among the three groups (P＞0.05). After treatment, the patients were followed up every month for 6 months. The changes of BCVA, CRT and the incidence of adverse reactions were observed during follow-up. One-way ANOVA and Kruskal-Wallis H test were used to compare the differences. ResultsDuring the follow-up period, compared with the baseline, the BCVA of the eyes in the initial combination treatment group, DEX treatment group and ranibizumab treatment group were significantly improved (Z=110.970, 90.359, 207.303), and CRT was significantly decreased (F=107.172, 88.418, 61.040), the difference was statistically significant (P＜0.01). At 1, 2, 3, 4, 5, and 6 months after treatment, there were significant differences in the mean changes in BCVA between the initial combined treatment group, DEX treatment group, and ranibizumab treatment group (χ2=34.522, 29.570, 14.199, 7.000, 6.434, 6.880; P＜0.05); 1, 2, 3, and 6 months after treatment, the differences were statistically significant (F=4.313, 7.520, 3.699, 3.152; P＜0.05). The time required to improve BCVA by 0.1 logMAR units in the initial combination treatment group, DEX treatment group, and ranibizumab treatment group was 5.73 (3.21, 8.48), 9.97 (6.29, 18.78), and 20.00 (9.41, 37.89) d, respectively; The time required for CRT to drop to 300 μm was 24.31 (21.32, 26.15), 29.42 (25.65, 31.37), and 29.17 (25.28, 36.94) d, respectively. The BCVA improvement of 0.1 logMAR unit and the time required for CRT to decrease to 300 μm in the eyes of initial combined treatment group were shorter than those in the eyes of DEX treatment group and the ranibizumab treatment group, and the differences were statistically significant (Z=－3.533, －4.445, －3.670, －4.030; P＜0.01). Different BRVO treatment groups: 1, 2, 3, 5, and 6 months after treatment, the mean BCVA changes were significantly different (χ2=24.989, 21.652, 11.627, 7.054, 9.698; P＜0.05); CRVO was different treatment group: 1 and 2 months after treatment, there were significant differences in mean BCVA changes (χ2=11.137, 9.746; P＜0.05). Two months after treatment, there were significant differences in CRT changes between BRVO and CRVO groups with different treatment regimens (F=3.960, 3.722; P＜0.01). The time required to improve BCVA by 0.1 logMAR unit in the eyes of BRVO and CRVO combined treatment group was shorter than that in the eyes of BRVO, CRVO DEX treatment group and the BRVO, CRVO ranibizumab treatment group, and the differences were statistically significant (BRVO: Z=－2.687, －3.877; P＜0.05; CRVO: Z=－2.437, －3.575; P＜0.05). The time required for CRT to drop to 300 μm in the CRVO combined treatment group was significantly shorter than that in the CRVO DEX treatment group and the CRVO ranibizumab treatment group, and the difference was statistically significant (F=6.910, P＜0.010); there was no statistically significant difference between the different BRVO treatment groups (F=1.786, P＞0.05). The number of re-treated eyes in the initial combined treatment group and DEX treatment group was less than that in the ranibizumab treatment group, and the difference was statistically significant (χ2=18.330, 7.224; P＜0.05). The retreatment interval of the eyes in the initial combined treatment group was significantly longer than that in the DEX treatment group and the ranibizumab treatment group, and the difference was statistically significant (P＜0.01). There was no significant difference in the incidence of intraocular hypertension among the initial combined treatment group, DEX treatment group and ranibizumab treatment group (χ2=0.058, P＞0.05). ConclusionsThe new strategy of initial combination therapy with DEX and ranibizumab in the treatment of RVO-ME has a better short-term effect. Compared with the monotherapy group, the retreatment interval is shorter, the visual and anatomical benefits are faster, the efficacy lasts longer, and the safety is better.
ObjectiveTo explore the effect of Kaempferol on bone microvascular endothelial cells (BMECs) in glucocorticoid induced osteonecrosis of the femoral head (GIONFH) in vitro. MethodsBMECs were isolated from cancellous bone of femoral head or femoral neck donated voluntarily by patients with femoral neck fracture. BMECs were identified by von Willebrand factor and CD31 immunofluorescence staining and tube formation assay. The cell counting kit 8 (CCK-8) assay was used to screen the optimal concentration and the time point of dexamethasone (Dex) to inhibit the cell activity and the optimal concentration of Kaempferol to improve the inhibition of Dex. Then the BMECs were divided into 4 groups, namely, the cell group (group A), the cells treated with optimal concentration of Dex group (group B), the cells treated with optimal concentration of Dex+1 μmol/L Kaempferol group (group C), and the cells treated with optimal concentration of Dex+5 μmol/L Kaempferol group (group D). EdU assay, in vitro tube formation assay, TUNEL staining assay, Annexin Ⅴ/propidium iodide (PI) staining assay, Transwell migration assay, scratch healing assay, and Western blot assay were used to detect the effect of Kaempferol on the proliferation, tube formation, apoptosis, migration, and protein expression of BMECs treated with Dex. ResultsThe cultured cells were identified as BMECs. CCK-8 assay showed that the optimal concentration and the time point of Dex to inhibit cell activity was 300 μmol/L for 24 hours, and the optimal concentration of Kaempferol to improve the inhibitory activity of Dex was 1 μmol/L. EdU and tube formation assays showed that the cell proliferation rate, tube length, and number of branch points were significantly lower in groups B-D than in group A, and in groups B and D than in group C (P<0.05). TUNEL and Annexin V/PI staining assays showed that the rates of TUNEL positive cells and apoptotic cells were significantly higher in groups B-D than in group A, and in groups B and D than in group C (P<0.05). Scratch healing assay and Transwell migration assay showed that the scratch healing rate and the number of migration cells were significantly lower in groups B-D than in group A, and in groups B and D than in group C (P<0.05). Western blot assay demonstrated that the relative expressions of Cleaved Caspase-3 and Bax proteins were significantly higher in groups B-D than in group A, and in groups B and D than in group C (P<0.05); the relative expressions of matrix metalloproteinase 2, Cyclin D1, Cyclin E1, VEGFA, and Bcl2 proteins were significantly lower in groups B-D than in group A, and in groups B and D than in group C (P<0.05). Conclusion Kaempferol can alleviate the damage and dysfunction of BMECs in GIONFH.
ObjectiveTo observe the changes of microstructure of rats'sciatic nerves with non-freezing cold injury after treated with dexamethasone. MethodsTwelve male Wistar rats were randomly divided into cooling group and treating group.Unilateral sciatic nerves of the rats in the cooling group received cooling treatment with 3-5℃ for 2 hours;while unilateral sciatic nerves of the rats in the treating group received cooling treatment with 3-5℃ for 2 hours and underwent the celiac injection with dexamethasone in addition.The other sciatic nerves were exposed,as the control.The bilateral sciatic nerves of rats in each group were harvested after 24 hours.The microstructure of nerves was examined under the light microscope and electron microscope. ResultsLight microscopic examination revealed extensive myelinated fibre degeneration in form of giant empty axons or shrunken dark axons on the first day after cooled.And the endoneurial capillary lumen was narrowed because of swollen endothelial cells.After the treatment,myelinated fibre degeneration was still similar to that before the treatment,but the endoneurial capillary lumen and endothelial cells were normal.By electron microscopy,sciatic nerves showed extensive myelinated fiber degeneration,and swollen endothelial cells.But unmyelinated fibers and tight junction were preserved on the first days after cooled.Aggregated red cells and platelet thrombus were not found.After the treatment,myelinated fibre degeneration was still similar to that before the treatment.Unmyelinated fibers and tight junction were preserved.The endoneurial capillary lumen and endothelial cells were normal. ConclusionAfter the treatment,the damage of endoneurial capillary had improved,but myelinated fiber degeneration was similar to that before the treatment.It suggested that dexamethasone may only improve the vascular system in non-freezing cold injury of sciatic nerve.
ObjectiveTo study the effect of transforming growth factor β3 (TGF-β3), bone morphogenetic protein 2 (BMP-2), and dexamethasone (DEX) on the chondrogenic differentiation of rabbit synovial mesenchymal stem cells (SMSCs). MethodsSMSCs were isolated from the knee joints of 5 rabbits (weighing, 1.8-2.5 kg), and were identified by morphogenetic observation, flow cytometry detection for cell surface antigen, and adipogenic and osteogenic differentiations. The SMSCs were cultured in the PELLET system for chondrogenic differentiation. The cell pellets were divided into 8 groups: TGF-β3 was added in group A, BMP-2 in group B, DEX in group C, TGF-β3+BMP-2 in group C, TGF-β3+DEX in group E, BMP-2+DEX in group F, and TGF-β3+BMP-2+DEX in group G; group H served as control group. The diameter, weight, collagen type II (immuohistochemistry staining), proteoglycan (toluidine blue staining), and expression of cartilage related genes [real time quantitative PCR (RT-qPCR) technique] were compared to evaluate the effect of cytokines on the chondrogenic differentiation of SMSCs. Meanwhile, the DNA content of cell pellets was tested to assess the relationship between the increase weight of cell pellets and the cell proliferation. ResultsSMSCs were isolated from the knee joints of rabbits successfully and the findings indicated that the rabbit synovium-derived cells had characteristics of mesenchymal stem cells. The diameter, weight, collagen type II, proteoglycan, and expression of cartilage related genes of pellets in groups A-F were significantly lower than those of group G (P<0.05). RT-qPCR detection results showed that the relative expressions of cartilage related genes (SOX-9, Aggrecan, collagen type II, collagen type X, and BMP receptor II) in group G were significantly higher than those in the other groups (P<0.01). Meanwhile, with the increase of the volume of pellet, the DNA content reduced about 70% at 7 days, about 80% at 14 days, and about 88% at 21 days. ConclusionThe combination of TGF-β3, BMP-2, and DEX can make the capacity of chondrogenesis of SMSCs maximized. The increase of the pellet volume is caused by the extracellular matrix rather than by cell proliferation.