1.Anti-oxidative stress effects of miR-125b on lens epithelial cells and its mechanism
Xiang LI ; Ying ZHANG ; Yalin HUANG ; Zhong WU ; Haoyi GUO
Chinese Journal of Experimental Ophthalmology 2021;39(4):280-288
Objective:To investigate the anti-oxidative stress effects of microRNA 125b (miR-125b) on lens epithelial cells (LECs) and its possible mechanism.Methods:Twenty-four anterior capsule specimens were collected from 24 eyes of 24 age-related cataract patients during phacoemulsification and 20 normal anterior capsule specimens were obtained from 20 eyes of 20 donors in Henan Eye Hospital from July 2018 to March 2019 under the approval of a Medical Ethics Committee of Henan Eye Hospital (No.YKYY20193151).The reverse transcription PCR and Western blot assay were employed to detect and compare the relative expression levels of miR-125b and nuclear factor E2-related factor 2 (Nrf2) in different specimens.The human lens epithelial cell line HLEB-3 was divided into control group and oxidative stress model group.The oxidative stress models were established by coculture with different concentrations (100, 200, 400 μmol/L) of H 2O 2 for 24 hours, and the cells were cultured with normal medium without H 2O 2 in the control group.The reactive oxygen species (ROS) content was detected by DCFH-DA fluorescent probe, and the activities of total-antioxidative capability (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) as well as malondialdehyde (MDA) concentration were detected by ELISA, and compared among the groups.The expression levels of miR-125b and Nrf2 were detected by reverse transcription PCR and Western blot assay, respectively.The cells were transfected with miR-125b mimics, miR-125b control and miR-125b inhibitor for 24 hours, respectively, and ROS content was detected by DCFH-DA fluorescent probe and T-AOC, SOD and GSH-Px activities as well as MDA concentration were detected by ELISA and compared among different transfected groups.A dual luciferase reporter assay was used to assess an association between miR-125b and Nrf2.The expression level of Nrf2 protein was detected by Western blot assay and the expression levels of Nrf2 and Keap1 were assayed and located by immunofluorescence double staining. Results:The relative expression levels of miR-125b and Nrf2 in the normal lens anterior capsule specimens were 0.21±0.03 and 0.27±0.06, which were significantly lower than 0.89±0.05 and 0.84±0.12 in the cataract specimens, respectively ( t=15.355, P<0.05; t=18.647, P<0.05).The relative expression levels of miR-125b and Nrf2 were significantly increased in various H 2O 2 treated groups in comparison with the control group and were gradually elevated with the increase of H 2O 2 concentration (all at P<0.05).Compared with the control group, the T-AOC, SOD and GSH-Px activities were reduced, and ROS content and MDA concentration were significantly ascended (all at P<0.05).Compared with the miR-125b control group, the T-AOC, GSH-Px and SOD activities were increased, and ROS content and MDA concentration were decreased in the miR-125b mimics group (all at P<0.05).In addition, the T-AOC, GSH-Px and SOD activities were significantly weakened, and ROS content and MDA concentration were significantly increased in the miR-125b inhibitor group in comparison with the miR-125b control group (all at P<0.05).Dual luciferase reporter assay showed that miR-125b targeted to the expression of Nrf2 in the H 2O 2 model cells.The fluorescence of Nrf2 in the cytoplasm was the strongest with more nuclear transfer in the miR-125b mimics group, and the expression intensity of Keap1 in the cytoplasm was weaker.The expression of Nrf2 was the weakest with less nuclear transfer in the miR-125b inhibitor group, and the expression level of Keap1 in the cytoplasm was stronger. Conclusions:MiR-125b can enhance the anti-oxidative stress of LECs in age-related cataractous eyes probably by upregulating the expression of Nrf2 and activating the Keap1/Nrf2 signaling pathway.
2.Clinical characteristics of elderly patients with coronavirus disease 2019
Zhigang CHANG ; Xin CHU ; Yongke ZHENG ; Yigang ZHONG ; Li WEN ; Ping ZENG ; Guifang ZHANG ; Chunyi FU ; Xunliang TONG ; Yunfei LONG ; Jing LI ; Aihua LIU ; Yalin LIU ; Huan XI
Chinese Journal of Geriatrics 2021;40(2):147-152
Objective:To investigate the clinical characteristics of elderly patients with coronavirus disease 2019(COVID-19), in order to provide scientific evidence for the diagnosis and treatment of COVID-19 in elderly patients.Methods:Clinical data of 102 patients with COVID-19 admitted to the B11 East Ward of the Zhongfaxincheng campus and the E1-3 ward of the Guanggu Campus of Tongji Hospital affiliated to Huazhong University of Science and Technology in Wuhan from 1 February 2020 to 28 February 2020 were retrospectively collected and analyzed.Patients were categorized into 2 groups: the elderly group(≥60 years old)and the young and middle-aged group(<60 years old). Differences in epidemiological features, demographics, clinical symptoms, laboratory results and imaging findings between the two groups were retrospectively analyzed.Results:Among 102 patients with COVID-19, 58 were in the elderly group(≥60 years old), with a median age of 67.0(63.8, 71.0)years old, and 44 in the young and middle-aged group(<60 years old), with a median age of 47.5(38.0, 51.8)years old.There was no significant difference in gender ratio between the two groups( χ2=0.033, P=0.855). Of 102 patients, 42.0%(21/50)had close contact with an infected person, 14.0%(7/50)were from infection clusters, and 18.0%(9/50)had suspected hospital-acquired infections.Fever and cough remained the most common symptoms, but gastrointestinal symptoms such as nausea, poor appetite, diarrhea and muscle cramps were also warning signs.Fatigue and cough were the most common presenting symptoms in elderly male patients.Bilateral patchy infiltrates(57.9%, 22/38)and ground-glass opacities(42.1%, 16/38)were the main imaging features and 42.1%(16/38)patients had multiple areas of the lungs involved.Over 50% patients had increased levels of blood glucose, D-dimer, fibrinogen, C-reactive protein, procalcitonin, multiple cytokines and neutrophil-to-lymphocyte ratio, as well as decreased levels of albumin, hemoglobin, hematocrit, lymphocytes and serum calcium.Compared with the young and middle-aged group, the elderly group had higher rates of abnormality in levels of D-dimer and serum calcium( χ2=7.067 and 4.166, P=0.008 and 0.041). Conclusions:Fever and cough are the most common symptoms in elderly patients with COVID-19.Elderly patients with COVID-19 have multiple abnormalities in clinical laboratory test results, which show a certain level of specificity compared with young and middle-aged patients.
3.Repair of open compound lower extremity wounds
Jingmin ZHU ; Tianzhi HAO ; Gang LU ; Zhi ZHOU ; Zhifu SUI ; Jingjie LIU ; Xiaojun TU ; Jinchao LUO ; Baoyu FAN ; Chuangong CHEN ; Yalin ZHONG
Chinese Journal of Trauma 2008;24(6):415-418
Objective To explore the repair of the open compound wounds in lower extremities caused by multiple factors. Methotis Transplantation of cutaneous.musculo-cutaneous or greater omentum flaps were applied to 155 patients of open compound lower extremity wounds. Results The wound healing rate following first operation was 50% and that following two operations was 14.8%.While the wounds were healed in 7.7% of patients after three operations. Conclusion Transplantations of cutaneous,musculo-cutaneous or greater omentum flaps ale effective to repair and reconstruct the open compound lower extremity wounds.
4.Analysis of Risk Factors of SkinAdverse Reactions by Intravenous Injection of Iodine ContrastAgent
Yanyan LI ; Yifan CHENG ; Yalin YANG ; Zhi’e FANG ; Dan HUANG ; Yunyi LI ; Mingchun HUANG ; Lijuan ZHONG ; Xinjie ZHOU ; Junda WANG
China Pharmacy 2020;31(18):2252-2258
OBJECTIVE:To investiga te main risk factors for adverse drug reactions (ADR)of skin by intravenous injection of iodine contrast agent. METHODS :From Jan. 2009 to Apr. 2020,the patients suffering from skin ADR after enhanced CT with iodine contrast agent were collected from our hospital. The basic information ,laboratory test results before using iodine contrast agent and ADR related information were collected through hospital information system (HIS). The use of iodine contrast agent ,main manifestations of skin ADR and drug combination were analyzed statistically. Taking the sex ,age,body mass index (BMI),the dosage of iodine contrast agent ,length of stay ,laboratory examination ,tumor history ,basic disease ,allergy history ,drinking history as independent variables ,the incidence of skin ADR related to iodine contrast agent was analyzed by single factor analysis ,and the variables with statistically significant were selected for multivariate Logistic stepwise regression analysis. RESULTS :There were 157 cases of skin ADR ,involving 79 males(50.3%)and 78 females(49.7%). The age ranged from 19 to 86 years old ,being(52.68± 18.73)years old in average. BMI was 14.6-40.7 kg/m2,being(22.5±3.7) kg/m2. 67 cases(42.68%)were treated with iprodione ,34 cases(21.66%)with iodixanol ,31 cases(19.74%)with iohexol and 25 cases(15.92%)with iopamidol ;the dose of iodine contrast agent were 50-100 mL,being(73.06±13.29)mL in average. There was no significant difference among different dosage of 4 kinds of iodine contrast agents (P≤0.05). Among 4 kinds of iodine contrast agents ,the incidence of skin ADR induced by iopromide was the highest(0.197%). The skin ADR related to iodine contrast agent was mainly acute (89.2%),the severity was mild (75.2%),and urticaria(38.9%)was the most common. After symptomatic treatment ,135 cases were cured ,13 cases were improved and 9 cases were not improved. Among the patients with iodine contrast agent related skin ADR ,the incidence of ADR induced by combined use of anti infective drugs was the highest (33.1%);however,the combined use of anti-tumor drugs was the main cause of severe skin ADR. The length of stay {11~20 d[OR=1.21,95%CI(1.07,1.20),P=0.042]、21~30 d[OR=1.39,95%CI(1.12,1.52),P=0.035]、31~40 d[OR=1.15,95%CI(1.03,1.37),P=0.008]、>40 d[OR=1.33,95%CI(1.28,1.53),P=0.003]},respitatory and circulatory system tumor history[OR =1.51,95%CI(1.35,1.61),P=0.037],injection allergy history[OR =1.50,95%CI(1.37,1.59),P=0.005] can significantly increase the incidence of iodine contrast agent related skin ADR. CONCLUSIONS :The main manifestation of skin ADR related to iodine contrast agent was urticaria. The main risk factors of skin ADR related to iodine contrast agent were length of stay (> 10 d),respiratory and circulatory system tumor history and injection allergy history.
5. Regulation and its mechanism of MeCP2 on biological behavior and epithelial-mesenchymal transition of LECs
Chao NIU ; Zhong WU ; Yalin HUANG ; Ying ZHANG ; Yingfei WANG ; Xiaohua LI ; Wenlong LU
Chinese Journal of Experimental Ophthalmology 2020;38(1):32-37
Objective:
To investigate the role of methyl-CpG-binding protein 2 (MeCP2) in the regulation and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) and its possible mechanism.
Methods:
Human LEC lines (SRA01/04) were divided into MeCP2-mimic group, MeCP2-NC group and small interferening RNA-MeCP2 (si-MeCP2) group, and MeCP2 analog plasmid, blank plasmid and MeCP2 si-RNA plasmid was used respectively to transfect the cells.The expression of MeCP2 mRNA in the cells was detected by using real-time PCR 24 hours after transfection.At 48 hours after transfection, the migration rate of the cells was evaluated by scratching test, and the expression of Wnt3a protein in the cells was detected by immunofluorescence stainning.The relative expressions of β-catenin, E-cadherin, Vimentin, matrix metallo proteinase (MMP)-9, MMP-7 and secreted frizzled-related protein 5 (SFRP5) proteins in the cells were detected by Western blot.
Results:
After 24 hours of transfection, the relative expression of MeCP2 mRNA in the cells was significantly different among the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group (