Objective:To study the therapeutic effect and mechanism of autologous platelet-rich plasma (PRP) on rabbit traumatic optic neuropathies (TON) and retina.Methods:Forty adult New Zealand white rabbits were selected to establish the optic nerve clamp injury model in their right eyes.According to the random number table method, 36 New Zealand white rabbits with effective model were randomly divided into model control group, normal saline control group and PRP group, 12 for each group.Another 12 healthy rabbits served as the normal control group.Rabbit autologous blood was collected to prepare PRP.The retrobulbar 20 μl PRP/20 μl saline solution injection was administered every two days near the injury after modeling according to grouping.The injection was carried out for 10 times.There was no other interference administrated to the model control group except the normal anti-infective treatment.No interference was given to the normal control group.At 30 and 60 days after modeling, the eyeballs and optic nerves of right eyes were harvested through sacrificing the animals by anesthetic overdose, three eyes for each time.Histopathological assessments were performed to observe the morphological changes of retina and optic nerve, and to evaluate the changes of retinal ganglion cells (RGCs) density and retinal nerve fiber layer (RNFL) thickness.Immunohistochemistry was used to assess the expressions of apoptosis factors caspase-3 and B cell lymphoma-2(Bcl-2). Quantitative real-time PCR and Western blot were used to detect the mRNA and protein expressions of brain-derived neurotrophic factor (BDNF) and growth associated protein-43 (Gap-43). This study protocol was approved by the Experimental Animal Ethics Committee of Wuhan University (No.E2019072805). The use and care of animals complied with ARVO statement.Results:The thickness of RNFL and number of RGCs at 30 days and 60 days after modeling were (6.60±1.16) μm, (6.89±1.21) μm, (13.00±1.00)/field of vision, (20.00±2.65)/field of vision in the PRP group, respectively, and were (4.80±0.43)μm, (2.18±0.23)μm, (6.33±0.58)/field of vision, (10.33±1.53)/field of vision in the model control group, respectively.The number of RGCs in the PRP group at 60 days was higher than that at 30 days after modeling, the number of RGCs in the PRP group was higher than that in the model control group, the thickness of RNFL in the PRP group was higher than that in the model control group; and the differences were statistically significant (all at P<0.05). At 30 and 60 days after modeling, the positive expression A value of caspase-3 protein in the normal saline group and model control group were higher than those in the normal control group and PRP group, while the positive expression A value of Bcl-2 protein in the PRP group was higher than those in the model control group and normal saline group, and the differences were statistically significant (all at P<0.05). The mRNA level and protein content of BDNF and Gap-43 in the retina and optic nerves at 30 days and 60 days after modeling in the PRP group were higher than those in the model control group, and the differences were statistically significant (all at P<0.05), but the mRNA and protein expression levels of BDNF and Gap-43 in different tissues in the PRP group at 60 days after modeling were lower than those at 30 days after modeling ( P<0.05). Conclusions:PRP can effectively inhibit the apoptosis of RGCs and the secondary injury of the retina after optic nerve injury, promote cell anti-apoptosis effect of RGCs, thereby retard the damage of the retina and optic nerve after TON, and also promote the repair of optic nerve and retina through upregulating the expression of nerve growth factors.

Objective To explore the image characteristics and clinical application of the optical coherence tomography (OCT) in patients with epiretinal membranes of the macular(ERMM). Methods 38 patients, who were diagnosed as or suspected as ERMM, were examined with OCT before and after operation from November, 2002 to October, 2003 in our hospital. Results Macular epiretinal membranes were visible on OCT as high reflective tissues, which were thin or thick, and contiguous to or anterior to the retinal surface. In most fovea, the depth decreased and the thickness increased. ERMM disappeared after operation. Conclusion OCT can display the macular epiretinal membrane and the pathological changes of macular tissues before and after operation. OCT can provide accurate information on the clinical diagnosis and operative efficacy of ERMM.

This study investigated the effect of etoposide, an anticancer chemotherapy drug, on B7-H1 expression in retinoblastoma (Rb) cells and the role of miR-513a-5p in the process. Rb cells were divided into control and etoposide groups. In the etoposide group, cells were treated with etoposide at different concentrations (2.5, 5, 10, 20 and 40 μg/mL) for 24 h. Those given no treatment of etopside served as controls. Reverse transcription polymerase chain reaction (RT-PCR), fluorescence quantitative PCR and flow cytometry were performed to measure the mRNA and protein expression of B7-H1 in Rb cells. The mRNA expression of miR-513a-5p in Rb cells before and after etoposide treatment was also detected by fluorescence quantitative PCR. The miR-513a-5p mimics and the miR-513a-5p inhibitor were transfected into Rb cells separately, and fluorescence quantitative PCR and flow cytometry were used to detect the effect of the miR-513a-5p mimics or inhibitor on B7-H1 expression. TargetScan5.2 was employed to predict the miR-513a-5p binding sites in the 3'-untranslated region of B7-H1 mRNA. Luciferase reporter plasmids carrying this site were prepared and transfected into Rb cells and luciferase activity analyzed. The results showed that etoposide stimulated the mRNA and protein expression of B7-H1 in Rb cells, which reached a maximal level after treatment with 5 μg/mL etoposide (P<0.05). However, miR-513a-5p expression was decreased in Rb cells after etoposide treatment. When the miR-513a-5p inhibitor was added, B7-H1 expression was increased with the concentration of the miR-513a-5p inhibitor (P<0.05). Moreover, B7-H1 expression was decreased gradually with the concentration of the miR-513a-5p mimics increased (P<0.01). Additionally, the miR-513a-5p mimics were found to inhibit the luciferase activity. It was concluded that etoposide can promote B7-H1 expression in Rb cells, which may be associated with chemoresistance. The promoting effect of etoposide on B7-H1 expression can be reversed by miR-513a-5p mimics. MiR-513a-5p inhibits the mRNA and protein expression of B7-H1 via binding to the 3'-UTR of B7-H1 mRNA.

Objective To investigate the clinical and pathological characteristics of IgA nephropathy (IgAN) in association with active tubular interstitial lesions. Methods 148 patients who were diagnosed as IgAN by renal biopsy and admitted to Zhejiang Provincial People's Hospital from March 2014 to December 2014 were enrolled. They were divided into IgAN with active tubular interstitial lesions group (IgAN?ATIL group, 23 patients) and IgAN without active tubular interstitial lesions group (control group, 125 patients). Clinical and pathological characteristics were retrospectively analyzed. Multivariate logistic regression analysis was used to analyze the influence factors. Results The prevalence of ATIL in 148 IgAN patients was 15.5%. IgAN?ATIL group showed an older average age and more higher proportion of medication history (antibiotics, diuretics, nonsteroidal anti ?inflammatory drugs, etc) than those in control group. There were significant differences in Alb, eGFR, Scr, BUN, 24?hour urinary protein quantity, urinary NGAL and urinary RBC count between two groups (P<0.05, respectively). A moderate of tubulointerstitial lesions of IgAN?ATIL group was shown, while the control group was mainly mild lesions. Multivariate logistic regression analysis showed that age, medication history and the urinary NGAL level were independent risk factors of IgAN ? ATIL. Conclusions IgAN patients with active tubular interstitial lesions had more severe clinical manifestations and chronic interstitial lesions. The age, medication history (antibiotics, diuretics, nonsteroidal anti?inflammatory drugs, etc) and the urinary NGAL level were independent risk factors of IgAN?ATIL.

Objective To evaluate the effect of bispectral index (BIS) and muscle relaxation monitoring on robot-assisted laparoscopic radical prostatectomy in elderly patients.Methods One hundred elderly patients (aged 65-80 years,ASA Ⅰ or Ⅱ) who underwent robot-assisted laparoscopic radical prostatectomy were randomly allocated into BIS and muscle relaxation monitoring group (group AA,n=50) and control group (group AC,n=50).In group AA,propofol was infused to achieve the BIS value of 45-55,and we monitored the muscle relaxation to conduct closed-loop infusion of cisatracurium.In group AC,we regulated the depth of anesthetic with the patients` vital signs according to anesthetists` experience.Mean arterial pressures (MAP),heart rates (HR),airway platform pressure (Pplat),and airway peak pressure (Ppeak) were recorded at following time points: before anesthesia induction (T0),after anesthesia induction (T1),10 min (T2),60 min (T3) after artificial pneumoperitoneum,and the end of operation (T4).We recorded dosage of propofol,cisatracurium,sufentanil,remifentanil,vasoactive agent,extubation time and PACU stay time.Results At T1,T2 and T4,the MAP and HR in group AC were significantly higher than those in group AA (P<0.05);at T3,MAP in group AC were apparently lower than those in group AA (P<0.05).Compared with T0,MAP and HR in group AC were significantly increased at T1,T2 and T4(P<0.05),MAP in group AC were obviously reduced at T3 (P<0.05),MAP and HR in group AC were also fluctuated obviously at different time points.MAP and HR in group AA at each point had no statistically significant difference.Compared with T1,Pplat and Ppeak in the two groups were significantly increased at T2-T4 (P<0.05).Pplat and Ppeak in grpup AC were higher than those in group AA at T2,T3 (P<0.05).Compared with group AC,the dosages of propofol and cisatracurium were less in group AA.The postoperative extubation time and PACU stay time were shorter in group AA.Conclusion BIS and muscle relaxation monitoring in robot-assisted laparoscopic radical prostatectomy can effectively stablize hemodynamics,reduce airway pressure fluctuation and the dosage of anesthetics.It also shortens the extubation time and the PACU stay time and improves the anesthesia recovery quality.

Background Proliferative vitreoretinopathy (PVR):no blood vessels,fibrous membrane cell proliferation occurs retinal surface.The development of specific mechanisms has not been completely clarified,but the retinal pigment epithelium(RPE) and platelet-derived growth factor(PDGF) for the past few years are the research hotspot.Objective To explore the effects of hypoxia on the production of PDGF-BB and proliferation in cultured human RPE (hRPE) cells.Methods In the experimental group,10,15,20,30,40 μmol/L CoCl2 were used to mimic hypoxia environment of hRPE cells,the control group did not add CoCl2 on cultured hRPE cells.By using reverse transcriptive PCR (RT-PCR) and enzyme linked immunosorbent assay(ELISA),the expression of PDGF-BB mRNA and protein was detected,and detected proliferation of hRPE cells by MTT.The cells were divided into 6 groups,different siRNAs were transfected into PDGF-BB siRNA1 group,PDGF-BB siRNA2 group,PDGF-BB siRNA3 group (because PDGF-BB has three different siRNAs,one of which is a valid siRNA),β-actin siRNA group,unrelated sequence siRNA group,only Lip2000 was plused in the blank control group.After transfection of 4-6 hours,except for blank control group,the other five groups were added to PDGF-BB mRNA and protein and its effect on the proliferation of hRPE cells obviously CoCl2 concentrations (15 μmol/L) simulated hypoxia environment of 24 hours,the detection of PDGF-BB mRNA and protein and hRPE cell proliferation rate.Results Neither PDGF-BB mRNA or protein was found in the blank control group.The production of PDGF-BB mRNA and protein in hRPE cells cultured by different concentrations were different,with significant differences among them (F=43.737,P＜0.01;F=54.612,P＜0.05),and the expressions of PDGF-BB mRNA and protein of 15 μmol/L CoCl2 group were more than other groups,with significant differences between the two groups (all at P＜0.05).MTT showed that PDGF-BB protein and hRPE cell proliferation rate in different concentions of CoCl2 groups were different,with significant differences among them(F=95.379,P＜0.01 ; F =63.375,P＜0.05),the expression of PDGF-BB protein was more and hRPE cell proliferation rate was higher in 15 μmol/L CoCl2group than other groups,with significant differences between the two groups (all at P＜0.05),postive correlation was found between PDGF-BB protein and hRPE cell proliferation rate (r =0.994,P＜0.05).The production of PDGF-BB mRNA and protein in the different siRNA transfected groups were different,with significant differences among them (F =156.330,125.650,both at P＜0.01),and the expressions of PDGF-BB mRNA and protein of PDGF-BB siRNA2 group were more than other groups,with significant differences between them(all at P＜0.05).MTT showed that PDGF-BB protein and hRPE cell proliferation rate in different siRNA transfectedgroups were different,with significant differences among them (F =73.131,98.564,both at P＜ 0.01).The expression of PDGF-BB protein was less and hRPE cell proliferation rate was lower in PDGF-BB siRNA2 group than other groups,with significant differences between them (all at P ＜ 0.05),postive correlation was found between PDGF-BB protein and hRPE cell proliferation rate (r =0.996,P＜0.05).Conclusions Proper hypoxia can induce the expression of PDGF-BB,PDGF-BB expression can significantly promote the proliferation of hRPE cells.In the transfection targeting PDGF-BB siRNA,PDGF-BB expression is suppressed,can effectively reduce the value of hRPE cell hyperplasia.

Objective To evaluate the effect of propofol on the endoplasmic reticulum stress-mediated apoptosis during focal cerebral ischemia-reperfusion (I/R) in rats.Methods Eighty adult male Sprague-Dawley rats,weighing 240-280 g,were randomly divided into 4 groups (n =20 each):shame operation group (group S) ; focal cerebral I/R group (group FCIR); propofol pretreatment group (group P); intralipid pretreatment group (group Ⅰ).Focal cerebral I/R was induced by 4 h middle cerebral artery occlusion followed by reperfusion.Propofol was infused at a rate of 12 mg· kg-1 · h-1 starting from 30 min before ischemia until 15 min of ischemia in group P,while intralipid was given instead of propofol in group I.Neurological deficit scores (NDSs) were measured at 6 h of reperfusion in 10 rats chosen from each group and the rats were then sacrificed.Their left brains were removed for determination of brain water content.The left 10 rats were sacrificed and their brains were immediately removed for determination of the expression of C/EBP homologous protein (CHOP),Bcl-2,and activated caspase-3 in the left hippocampi by Western blot.Results Compared with group S,NDSs and brain water content were significantly increased,the expression of CHOP and activated caspase-3 was up-regulated,and the expression of Bcl-2 was down-regulated in group FCIR,NDSs was increased in group P (P ＜ 0.05).Compared with group FCIR,NDSs and brain water content were significantly decreased,the expression of CHOP and activated caspase-3 was down-regulated,and the expression of Bcl-2 was up-regulated in group P,and no significant change was found in each parameter in group Ⅰ (P ＞ 0.05).Conclusion Propofol can reduce focal cerebral I/R injury through inhibition of the endoplasmic reticulum stress-mediated apoptosis in rats.

Objective:To investigate the effect of heat shock protein B8 (HspB8) downregulation on retinal ganglion cell (RGC) and retinal function in the mice model of optic nerve injury (ONC).Methods:Adeno-Associated Virus (AAV) 2 AAV2-shHspB8-GFP was constructed to knockdown HspB8. 66 adult male C57/BL6 mice were randomly divided into the control group, the ONC group, the AAV2-shHspB8 group, the ONC+AAV2-shHspB8 group, and the ONC+AAV2-GFP group. There were 10, 20, 16, 10 and 10 mice respectively, and both eyes were used as experimental eyes. Western blot was used to evaluate the expression of HspB8 on day 3 and 7 after ONC. By GFP immunofluorescence staining, the efficacy of AAV2-shHspB8-GFP transfer was accessed. Moreover, it was possible to identify functional and RGC survival differences between groups by optomotor response (OMR), dark adapted full-field flash electroretinogram (ff-ERG), oscillatory potentials (OPs), photopic negative response (PhNR) and retinal flat-mount RGC counting 5 days after ONC. Comparisons between two groups were made using Mann-Whitney U test, unpaired t-test, unpaired t-test with Welch’s correction, one-way ANOVA, and Bonferroni t test. Results:Compared with the control group, the expression of HSPB8 protein in the retina of mice in ONC3 group was significantly increased, and the difference was statistically significant ( F=43.63, P<0.01). Compared with the control group, the ONC group showed obviously lower visual acuity ( P<0.01), lower a-wave, b-wave, OPs, PhNR amplitude, longer b-wave latency ( P<0.05), and the survival rates of RGC in ONC3 group, ONC5 group and ONC7 group decreased in a time-dependent manner( F=384.90, P<0.01). Transfection of AAV2 efficiency was highest on 4 weeks after IVT. Besides, there was no significant differences between the control group and the AAV2-shHspB8 group on visual acuity, ff-ERG, OPs, PhNR and RGC survival ( P>0.05). In comparison of the control group, we found that RGC survival of the ONC5+AAV2-shHspB8 group was significantly elevated ( F=10.62, P<0.01). Conclusions:Expression of HspB8 on the retina can be induced by ONC. The investigation of RGC counting, visual acuity, and ff-ERG revealed that optic nerve injury destructed functionality of mice retina and resulted to RGC death ultimately. The Most crucial finding of this research is that HspB8 knockdown had a neuroprotective effect in RGC after ONC.

Objective:To investigate the inhibitory effects of resveratrol (Res) on the apoptosis of retinal neurons of diabetic rats and ARPE-19 cells induced by high glucose.Methods:Streptozotocin was intraperitoneally injected to establish a diabetes model in 25 adult male SD rats, and 20 successful models were randomized into diabetic model group and Res-administered group according to random number table method.Another 10 matched normal rats were served as normal control group.Res was intragastrically given to the rats in the Res-administered group with the dose of 40 mg/(kg·d), and an equivalent volume of normal saline solution was used in the same way in the diabetic model group and normal control group.The body weight and blood glucose level were measured on the 0th, 4th, 8th, 12th week of administration.The rats were sacrificed on the 12th week by over-anesthesia and the eyeballs were enucleated.The ultrastructure of the retinal ganglion cells (RGCs) were examined under the transmission electron microscope.The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) assay was to assess the apoptosis of retinal neurons.ARPE-19 cells were divided into normal control group, high glucose group and Res-treated group and cultured for 48 hours in medium containing 5.5 mmol/L glucose, 30 mmol/L glucose, 30 mmol/L glucose and 10 mol/L Res, respectively.Apoptosis rate was detected by a flow cytometry.The distribution and expression of bax and bcl-2 in the cells was detected by immunofluorescence and Western blot assay, respectively.This study protocol was approved by an Experimental Animal Ethic Committee of Hubei University of Science and Technology, and the use and care of the animals complied with the Statement of ARVO.Results:Compared with the diabetic model group, the body weight was increased at 4-12 weeks and the blood glucose level was lowered at 8-12 weeks of Res administration in the Res-administered group (both at P<0.01). The chromatin condensation, mitochondrial swelling and vacuolation in cytoplasm were obviously slight and the apoptosis rate was reduced in the Res-administered group in comparison with the diabetic model group.The apoptosis indexes of the retinal ganglion cell layer cells and inner nuclear layer cells in the Res-administered group were (18.36±3.37)% and (23.67±8.98)%, respectively, which were significantly lower than (83.91±9.8)% and (64.26±10.66)% in the diabetic model group (both at P<0.01). The apoptosis rate of the ARPE-19 cells in the normal control group, high glucose group and Res-treated group was (3.11±0.26)%, (11.41±1.06)% and (5.38±0.58)%, respectively, and the apoptosis rate in the Res-treated group was significantly lower than that in the high glucose group (all at P<0.01). Immunofluorescence assay showed that the fluorescence of bax in the cell nucleus of Res-treated group was obviously enhanced in comparison with the normal control group and weaker in comparison with the high glucose group.The fluorescence of bcl-2 protein in the Res-interfered group was weaker in comparison with the normal control group and enhanced in comparison with the high glucose group.The relative expressions level of bax protein in the Res-treated group was 0.21±0.08, which was significantly higher than 0.15±0.06 in the normal control group and lower than 0.31±0.09 in the high glucose group (both at P<0.05). The relative expressions of bcl-2 protein was 0.66±0.25 in the Res-treated group, which was significantly lower than 0.80±0.14 in the normal control group and higher than 0.23±0.09 in the high glucose group (both at P<0.05). The bcl-2/bax ratio in the Res-treated group was significantly higher than that in the high glucose group ( P<0.01). Conclusions:Res can inhibit the apoptosis of RGCs of diabetic rats and high glucose-induced RPE cells in vitro.

This study investigated the effect of etoposide, an anticancer chemotherapy drug, on B7-H1 expression in retinoblastoma (Rb) cells and the role of miR-513a-5p in the process. Rb cells were divided into control and etoposide groups. In the etoposide group, cells were treated with etoposide at different concentrations (2.5, 5, 10, 20 and 40 μg/mL) for 24 h. Those given no treatment of etopside served as controls. Reverse transcription polymerase chain reaction (RT-PCR), fluorescence quantitative PCR and flow cytometry were performed to measure the mRNA and protein expression of B7-H1 in Rb cells. The mRNA expression of miR-513a-5p in Rb cells before and after etoposide treatment was also detected by fluorescence quantitative PCR. The miR-513a-5p mimics and the miR-513a-5p inhibitor were transfected into Rb cells separately, and fluorescence quantitative PCR and flow cytometry were used to detect the effect of the miR-513a-5p mimics or inhibitor on B7-H1 expression. TargetScan5.2 was employed to predict the miR-513a-5p binding sites in the 3'-untranslated region of B7-H1 mRNA. Luciferase reporter plasmids carrying this site were prepared and transfected into Rb cells and luciferase activity analyzed. The results showed that etoposide stimulated the mRNA and protein expression of B7-H1 in Rb cells, which reached a maximal level after treatment with 5 μg/mL etoposide (P<0.05). However, miR-513a-5p expression was decreased in Rb cells after etoposide treatment. When the miR-513a-5p inhibitor was added, B7-H1 expression was increased with the concentration of the miR-513a-5p inhibitor (P<0.05). Moreover, B7-H1 expression was decreased gradually with the concentration of the miR-513a-5p mimics increased (P<0.01). Additionally, the miR-513a-5p mimics were found to inhibit the luciferase activity. It was concluded that etoposide can promote B7-H1 expression in Rb cells, which may be associated with chemoresistance. The promoting effect of etoposide on B7-H1 expression can be reversed by miR-513a-5p mimics. MiR-513a-5p inhibits the mRNA and protein expression of B7-H1 via binding to the 3'-UTR of B7-H1 mRNA.
B7-H1 Antigen ; genetics ; Cell Line, Tumor ; Etoposide ; pharmacology ; Gene Expression ; drug effects ; genetics ; Humans ; MicroRNAs ; genetics ; Retinoblastoma ; genetics