Dendrobii Caulis Mixture-containing Serum Protects Mice from Podocyte Injury Induced by High Glucose / 中国实验方剂学杂志

ObjectiveTo investigate the effect and mechanism of Dendrobium mixture （DMix）-containing serum on high glucose-induced podocyte injury in mice. MethodThe MPC5 mouse glomerular podocytes were cultured in vitro， and the optimal glucose concentration for modeling， modeling time， and concentration of DMix-containing serum for administration were determined. The cells were classified into normal （5.5 mmol·L-1 glucose+10% blank serum）， model （30 mmol·L-1 glucose+10% blank serum）， DMix-containing serum （30 mmol·L-1 glucose+10% DMix-containing serum）， ferroptosis inhibitor （Fer-1， 30 mmol·L-1 glucose+10% blank serum+1 μmol·L-1 Fer-1） groups. The corresponding kits were used to measure the levels of Fe2+ and lactate dehydrogenase （LDH） in cells. Enzyme-linked immunosorbent assay was employed to determine the content of glutathione （GSH） and lipid peroxide （LPO） in cells. Fluorescence probe was used to measure the reactive oxygen species （ROS） level. Real-time fluorescence quantitative polymerase chain reaction and Western blotting were employed to determine the mRNA and protein levels， respectively， of Wilms' tumor-1 （WT-1）， desmin， long chain acyl-CoA synthase 4 （ACSL4）， and glutathione peroxidase 4 （GPX4） in podocytes. ResultCompared with the blank group， the intervention with 30 mmol·L-1 glucose for 48 h reduced podocyte viability （P<0.01）， and the 10% DMix-containing serum showed the most significant improvement in podocyte viability （P<0.01）. Compared with the normal group， the model group presented elevated levels of Fe2+， LDH， LPO， and ROS， lowered GSH level， up-regulated mRNA and protein levels of desmin and ACSL4， and down-regulated mRNA and protein levels of WT-1 and GPX4 （P<0.01）. Compared with the model group， the DMix-containing serum lowered the Fe2+， LDH， LPO， and ROS levels， elevated the GSH level， down-regulated the mRNA and protein levels of desmin and ACSL4， and up-regulated the mRNA and protein levels of WT-1 and GPX4 in podocytes （P<0.05， P<0.01）. ConclusionDMix-containing serum exerts a protective effect on high glucose-induced podocyte injury by inhibiting ferroptosis.

Macrophage-to-myofibroblast transition promotes pulmonary fibrosis occurred in LPS-induced acute lung injury of mouse models / 基础医学与临床

Objective To explore the impact of macrophage-to-myofibroblast transition(MMT)on pulmonary fibro-sis induced by acute lung injury by LPS.Methods Totally 21 male mice were randomly classified into 7 groups:control group,model group(LPS-PF)at different time points and intervention group of clodronate-liposomes(CL-LIP)treatement at different time points(n=3).Pulmonary fibrosis was identified by HE and Masson staining microscopy.The immuno-fluorescence technology was used for the evaluation of numbers of macrophage-to-myofi-broblast transition cells(MMT cell which co-expressed CD68 and α-SMA).Bone marrow-derived macrophages(BMDMs)were randomly classified into two group:control(Ctrl)group and TGF-β1-treated group induced by transforming growthfactor-β1.α-SMA,FN and Col1 were detected by RT-qPCR.The expression of α-SMA,Smad3 and p-Smad3 protein was evaluated by Western blot.Results At day 7,the Ashcroft score of lung tissue in LPS-PF mouse model was significantly increased when compared with the Ctrl group(P＜0.01);While the score signifi-cantly declined when the model was pretreated with CL-LIP(P＜0.05).As detected by immuno-fluorescence stai-ning,in CL-LIP group the number of CD68-positive cells co-labeled with α-SMA was obviously less then that of LPS-PF group of the corresponding time point(P＜0.01).When the BMDMs were stimulated by TGF-β1 at 24 h,48 h and 96 h respectively,a higher expression of α-SMA,FN,Col1,were found in TGF-β1-treated group than that in Ctrl group at the corresponding time point(P＜0.01).The expression of Smad3,p-Smad3 significantly higher in LPS-PF group(at both day 7 and day 10)and TGF-β1-treated group(at both 48 h and 96 h)as compared to cor-responding control group(P＜0.01).Conclusions MMT promotes pulmonary fibrosis induced by ALI via LPS.Smad3 is proved to be involved in the MMT process.

Inhibitory effect of exosome-loaded KV11 on corneal neovascularization via VDAC1 and autophagy / 中华实验眼科杂志

Objective:To investigate the effects and mechanism of exosome (EXO)-loaded kringle V11 (KV11) delivery on corneal neovascularization (CNV).Methods:KV11 was bound to the surface of endothelial cell-derived exosomes by using CP05, an EXO-targeting anchoring peptide, to produce EXO-KV11.The binding efficiency and optimal concentration ratio were determined using the Apogee flow system.A total of 100 8-week-old healthy male SPF grade SD rats were selected, 10 of which were randomly selected as a normal control group without any treatment.The CNV model was established by alkali burn in the other 90 rats, which were randomly divided into three groups, EXO-KV11 group, KV11 group, and normal saline group by the random number table method, with 30 rats in each group.Each group was injected subconjunctivally with 100 μl of EXO-KV11 (25 μg), KV11 (25 μg), or normal saline every other day from the first day after the alkali burn, respectively.The CNV of rats was observed on days 1, 4, 7, and 14 after alkali burn.The CNV area was calculated by ventricular perfusion with fluorescein isothiocyanate-dextran (FITC-dextran) and corneal angiography.The amount of CNV lumen was observed by hematoxylin and eosin staining.The distribution of CD31 in rat corneas was determined by immunohistochemical method.The expression levels of voltage-dependent anion channel 1(VDAC1), endoplasmic reticulum stress, autophagy and apoptosis-associated proteins were detected by Western blot.This study was approved by the Animal Ethics Committee of Peking University People's Hospital (No.20210019). All animal procedures complied with the regulations of the Vision and Ophthalmology Association and the Animal Protection and Use Committee of Peking University.Results:The optimal concentration ratio of KV11 to EXO was 4∶1 and the binding affinity reached up to 87.5% by Apogee flow cytometers.On days 7 and 14 after alkali burn, there were significant differences in CNV area among the four groups ( F=4.613, 15.590; both at P<0.05). On day 7 after alkali burn, the CNV area was smaller in EXO-KV11 group than in KV11 and normal saline groups, with statistically significant differences (both at P<0.05). On day 14 after alkali burn, the CNV area was smaller in EXO-KV11 and KV11 groups than in normal saline group, and smaller in EXO-KV11 group than in KV11 group, showing statistically significant differences (all at P<0.05). The results of quantitative analysis of corneal fluorescence mounts showed that the relative CNV fluorescence area of the normal saline group, KV11 group and EXO-KV11 group were (8.3±1.7)%, (5.2±1.6)%and (3.4±0.7)%, respectively, showing a statistically significant overall comparison difference ( F=11.735, P<0.01). The relative CNV fluorescence area was larger in KV11 and normal saline groups than in EXO-KV11 group, and larger in normal saline group than in KV11 group, showing statistically significant differences (all at P<0.05). On day 14 after alkali burn, massive neovascular lumens were observed in the matrix of the normal saline group.The number of neovascular lumens in KV11 group was smaller than that in normal saline group.The corneal structure appeared normal in EXO-KV11 group, and neovascular lumens were rare.Numerous CD31-positive cells were observed in the corneal stroma of the normal saline group, which formed into lumen structures.The number of lumens surrounded by CD31-positive cells in the corneal stroma was smaller in KV11 group than in normal saline group, and smaller in EXO-KV11 group than in KV11 group.There were significant differences in the relative expression levels of VDAC1, protein kinase R-like endoplasmic reticulum kinase (PERK), p62, cleaved caspase 3 among the four groups ( F=35.960, 8.947, 17.791, 101.168; all at P<0.01). The relative expression levels of VDAC1, PERK, p62, cleaved caspase 3 were higher in EXO-KV11 group than in KV11 and normal saline groups, showing statistically significant differences (all at P<0.001). There was no significant difference in the relative expression of microtubule-associated proteins 1A/1B light chain 3B (LC3B)Ⅱ/LC3BⅠ protein among all four groups ( F=0.445, P=0.727). Conclusions:EXO-KV11 can inhibit CNV more remarkably than KV11.EXO-KV11 inhibits CNV by promoting the expression of VDAC1 and PERK and suppressing the autophagic flux.

Extended internal limiting membrane peeling combined with releasing the edge for primary failed idiopathic macular hole surgery / 中华眼底病杂志

Objective:To explore the outcome of extended internal limiting membrane (ILM) peeling combined with releasing the edge technique for primary failed idiopathic macular hole (IMH) surgery.Methods:A retrospective analysis was performed. The data of 18 eyes of 18 IMH patients who were failed in primary surgery from August 2013 to June 2019 in Peking University People’s Hospital were enrolled in the study. Among them, 5 patients were males and 13 patients were females. The average age was 66.2±6.4 years. The BCVA were measured by ETDRS charts. The minimum macular hole size was measured on OCT B-scan image. The average preoperative BCVA and minimum macular hole size of primary surgery was 32.6±13.1 letters and 621.1±161.8 μm. The average preoperative BCVA and minimum macular hole size of second surgery was 34.4±12.3 letters and 499.0±148.6 μm. Average interval period of first and second surgery was 3.4±1.3 weeks. The surgical technique used in the reoperation included the extended ILM peeling combined with releasing the MH edges. The extended ILM peeling area ranged from 4 DD diameter to vascular arcades. The technique of releasing the macular hole edges was performed by using a silicone soft-tip extrusion cannula, with which tapping the edges softly or aspirated vacuum the edges concentricly. The average follow-up was 9.3±5.2 months. The clear OCT image can be obtained for confirming MH closure which was considered as the closure time in the first time. The comparison of preoperative and postoperative was performed by paired t-test. Results:The closure rate of second surgery was 94.4% (17/18), and average closure time was 2.1±1.0 weeks. Only 1 eye experienced the second surgical failure and received the third surgery to achieved macular hole closure. Average final follow-up BCVA was 51.5±13.0 ETDRS letters, with average improvement of 18.9 ETDRS letters (>3 lines) compared with preoperative BCVA of primary surgery. There was significantly statistical difference between the final BCVA and preoperative BCVA ( t=5.412, P <0.001). Eleven patients (61.1%) had 3 lines BCVA improvement, 15 patients (83.3%) had more than 1 line improvement, and 3 patients (16.7%) had on improvement. The final BCVA of patients significantly improved compared with preoperative BCVA of the second surgery ( t=7.595, P<0.001), with average improvement of 17.1±9.5 letters. Conclusion:The extended ILM peeling combined with releasing macular hole edges technique is effective to improve the closure rate and BCVA of primary failed IMH eyes.

Correlations between readiness for hospital discharge and self-efficacy in elderly patients with chronic obstructive pulmonary disease / 中华健康管理学杂志

Objective To investigate the level of readiness for hospital discharge in elderly patients with chronic obstructive pulmonary disease (COPD) and its correlation with patients'self-efficacy. Methods A questionnaire survey was administered to 240 elderly COPD patients admitted to the people's hospital of Wuhan University in Hubei Province from November 2017 to March 2018, using the Readiness for Hospital Discharge Scale and the Self-Efficacy for Managing Chronic Disease 6-Item Scale. Pearson correlations were used for analysis. Results Total scores for hospital discharge readiness were (6.13±0.89) and for self-efficacy were (3.42±1.46) in elderly COPD patients. Pearson correlation analyses indicated that there were significant positive correlations between the total scores and all dimensions of readiness for hospital discharge and self-efficacy in elderly patients with COPD (r=0.256-0.651, P<0.01). Conclusion The readiness for hospital discharge in elderly patients with COPD is still at a medium level, and the level of self-efficacy is low. Readiness for hospital discharge and self-efficacy are positively correlated in elderly patients with COPD. Health care workers should pay active attention to the readiness for hospital discharge in elderly patients with COPD so as to help them improve their self-efficacy, and promote disease recovery.

Application value of optical coherence tomography in predicting visual outcome after vitrectomy for idiopathic macular epiretinal membrane / 中华实验眼科杂志

Background Idiopathic macular epiretinal membrane (IMEM) shows a high incidence and affects visual function of patients.The primary management of IMEM is vitrectomy,but the study on prediction of visual acuity after operation by optical coherence tomography (OCT) is seldom.Objective This study was to evaluate the predicting ability of OCT for preoperative visual outcome for IMEM patients who received vitrectomy.Methods The clinical data of IMEM patients with vitrectomy from March 2009 to May 2013 were retrospectively analyzed in People's Hospital Peking University.Forty-nine eyes of 48 cases were divided into continuous group (19 eyes of 18 cases) and discontinuous group (31 eyes of 31 cases) based on whether the extraretinal layers were intact on OCT,including retinal pigment epithelium (RPE),inner and outer segment of photoreceptors (IS/OS) and external limiting membrane (ELM).Central foveal thickness (CFT) by high-resolution spectral-domain OCT and best corrected visual acuity (BCVA) (LogMAR) before surgery and 12 weeks after surgery were recorded and compared between the two groups,and the correlations of shift of CFT with the shift of LogMAR vision was analyzed by unitary linear regression to evaluate the value of OCT in predicting postoperative vision.Writen informed consent was obtained from each patient prior to any medical operation.Written informed consent was obtained from each patient before surgery.Results In 12 weeks after operation,reflective strips of RPE layer,IS/OS and ELM layer were intact in 18 eyes of the continuous group after vitrectomy.The number of eyes with discontinuous strip in RPE layer decreased to 1 eye after operation from 6 eyes before operation,and the number of eyes with IS/OS reflective strip discontinue decreased to 19 eyes after operation from 29 eyes before operation,and that with ELM layer discontinue decreased to 15 eyes after operation from before operation in the discontinuous group.The improvement of LogMAR BCVA was increased with the decline of CFT values with a negative correlation between them (R2 =0.298,B =0.001,P =0.000),and postoperative LogMAR visual acuity improved 1 line whenever CFT reduced by 100 μm.LogMAR BCVA was 0.4±0.19 before surgery and 0.36±0.21after surgery in the continuous group,showing a insignificant difference between them (t =0.876,P=0.393).In the discontinuous group,the postoperative IogMAR BCVA was significantly higher after operation (0.46±0.26) than 0.82 ±0.41 after operation (t =6.206,P =0.000).The increase value of LogMAR after vitrectomy was 0.3 in the discontinuous group,which was superior to 0.0 of the continuous group.Conclusions The continuity of extraretinal layers and CFT by OCT are closely associated with the improvement of vision after vitrectomy in the eyes with IMEM.Postoperative vision is better in IMEM eye with continuous retinal reflective strips before operation,and majority of IMEM eyes can improve vision after operation.Preoperative OCT seems to be an important diagnostic tool for the selection of patients who benefit or not from surgery to certain extent.

The clinical study on intravitreous injection of avastin for macular edema induced by branch retinal vein occlusion / 眼科研究

Background Branch retinal vein occlusion (BRVO)often leads to macular edema as the result of elevating intravitreal VEGF level,and avastin,a anti-VEGF drug,show a good effectiveness on macular edema secondary to BRVO.ObjectiveThis study attempts to evaluate the clinical efficacy of avastin on macular edema induced by BRVO.MethodsThis is a retrospective case-observation study.The clinical data of 39 eyes from 39 patients with macular edema induced by BRVO were included in this analysis.All of the patients received intravitreal injection of 1.25mg (0.05mL)avastin without other any therapy prior to the injection of avastin.This procedure followed the Declaration of Helsinki,and written informed consent was obtained from all the patients before and initial of any management.Clinical indexes included best-corrected visual acuity(BCVA),slit-lamp examination,intraocular pressure and stereoscopic biomicroscopy examination before injection and 3,6 and 12 weeks after initial injection.Optical coherence tomography (OCT),fundus photography,fluorescein fundus angiography(FFA)were performed prior to injection and 6,12 weeks after initial injection.The follow-up period was 3-20 months.ResultsThe mean BCVA was significantly improved at 3,6,12 weeks after injection in comparison with before injection (t=-6.039,-6.182,-4.189,all P=0.000).The mean CMT showed a statistically significantly decline at 6,12 weeks after injection in comparison with before injection(t=8.684,5.019,all P=0.000).No ocular or systemic adverse events were found after intravitreal injection of avastin during the follow-up duration.The numbers of visual acuity-improved eyes were significantly increased in the patients with disease course ≤1 month duration in comparison to ones with the course ≥1 month (P<0.05)in 3 weeks after injection.CMT was obviously decreased in 12 weeks after injection in comparison to before injection between with and without macular perfusion eyes (P<0.05).ConclusionIntravitreal injection of avastin is safe and effective for macular edema induced by BRVO,especially the patients with shorter course of disease.

Effects of levodopa on nerve growth factor in visual cortex of kitten / 眼科研究

Objective Nerve growth factor (NGF) is thought to be one of regulating factors of cellular function during the visual plasticity period. Levodopa is a neurotransmitter in central nerve system. Its mechanism of improving the visual function in amblyopia is unclear. Present study was to observe the role of levodopa in treating monocular deprived amblyopia and explore the effects of levodopa on the expression of NGF in visual cortex. Methods The animal models of monocular form deprived amblyopia were established in the right eyes of 12 4-week-old domestic cats by suturing the eyelid for 2 weeks and then the left eyes were reverse sutured. The 6 cats from models received 20 mg/kg of levodopa by oral administration. Other 6 normal cats were as normal controls. Pattern VEP (P-VEP) was recorded to check the visual acuity of the cats. The cell number density of NGF expression in visual cortex area 17 was detected and calculated by immunochemistry and expressed as positive number/field. Results The amplitude of P_(100) of P-VEP was significantly declined in monocular deprived eyes compared with fellow eyes and normal eyes in 2 weeks after monocular deprivation (P ＜0. 01) . However, after administration of levodopa, the amplitude of P_(100) recovered to normal 2 weeks later, presenting a insignificant difference between levodopa treating group and normal group (P＞ 0. 05). The latency of P_(100) was followed the same pattern between two groups. The NGF positive cells were decreased in monocular deprived group compared with normal group and levodopa group (80. 23 ± 9. 54 vs 111. 83 ± 7. 49, 80. 23 ± 9. 54 vs 118. 06 ± 12. 37, P ＜ 0. 01), but no significant difference in NGF positive cells was found between normal group and levodopa group (111.83 ±7. 49 vs 118. 06 ± 12. 37, P = 0. 94). Conclusion Levodopa is supposed to accelerate the recovery of visual function in amblyopic cat by enhancing the NGF expression in visual cortex area 17 during the critical period of visual development.

Ultrastructure collagen fibril organization in vitreous and the effect of plasmin on it / 中华眼底病杂志

Objective To reveal the fibrillar network in vitreous and the effect of plasmin on this network. Methods 20 vitreous gels of freshly slaughtered pigs were divided into 2 groups, the gels in first group were digested by 3 U plasmin (3 U/ml) at 37℃ for 24 hours respectively, the second group received the same PBS as control. After digestion, gels were fixed in neutral buffered formalin solution. Samples from vitreous base, cortex and the central region were observed by the technique of freeze etching electron microscopy. Results In vitreous collagen fibril network was in a three-dimensional array, collagen fibril density showed marked differences, central vitreous had the sparse fibril density, the cortex denser and the basal vitreous densest. After digestion by plasmin, the collagen fibrillar network was destructed. Conclusion Collagen fibrils in vitreous present spatial arrangement regularly, plasmin can lead to destruction of the fibrillar network.