1.Protective effect of Humanin on rotenone-induced dopamine neuron toxicity
Yaohui SHAN ; Qifu ZHANG ; Jin CHENG ; Feng YE ; Xi ZHANG ; Wenpei YU ; Xiaogang WANG ; Yuanpeng ZHAO ; Guorong DAN ; Mingliang CHEN ; Yan SAI
Journal of Army Medical University 2024;46(7):670-677
Objective To investigate the mechanism and protective effect of Humanin(HN)on rotenone(Rot)-induced toxic damage for dopamine neurons.Methods The Rot-poisened PC12 cell model was constructed,and the control group,the Rot poisening group,the HN pretreated Rot poisening group,and the HN treatment group were set up.ELISA was used to detect the content of HN inside and outside of Rot-infected cells,CCK-8 assay was used to detect cell viability,and ATP detection kit was used to detect the intracellular ATP content.Dichloro-dihydro-fluorescein diacetate(DCFH-DA)assay was used to detect the level of reactive oxygen species(ROS)in cells.Western blotting was performed to detect the expression level of mitochondrial autophagy regulatory proteins Pink1,Parkin,p62,LC3,mitochondrial biogenesis regulatory protein PGC1α,division/fusion regulatory proteins OPA1,MFN2,DRP1,p-DRP1 and antioxidant stress regulatory proteins Keap1 and Nrf2.HBAD-mcherry-EGFP-LC3 adenovirus transfected cells was used to observed the number of autophagosomes and autophagolysosomes.Results The results showed that the intracellular concentration of HN in PC12 in the Rot poisening group was significantly higher than that in the control group(P<0.05);Compared with the control group,the Rot poisening group had significantly decreased activity of PC12 cells,decreased ATP content and increased production of ROS.After the poisen of Rot in PC12 cells,the expression of Pink1 and p-Parkin,the ratio of LC3Ⅱ/LC3Ⅰ and the expression of p-DRP1 in mitochondrial fusion protein was increased,while the expression of p62,the expression of mitochondrial biogenesis protein PGC1 α,mitochondrial fusion proteins MFN2 and OPA1,and antioxidant stress proteins Keap1 and Nrf2 were decreased(all P<0.05).The number of autophagosomes and autophagolysosomes in PC12 cells in the Rot poisening group was higher than that in the control group(P<0.05),and HN pretreatment(20 μmol/L)could significantly improve the changes mentioned above caused by Rot poisening(P<0.05).Conclusion HN ameliorates Rot-induced toxic damage for dopamine neurons by inhibiting mitophagy and mitochondrial division and promoting mitochondrial biogenesis and fusion,and anti-oxidative stress.
2.Research progress of miRNA in asthma:airway inflammation and en-dotype
Bahu BAO ; Weiying LIU ; Yucai YE ; Guorong CHEN ; Jingzi SUN ; Aoyan HU
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(10):1194-1200
microRNA(miRNA)is a 22nt long sin-gle-stranded non-coding RNA that is involved in a variety of physiological and pathological processes.Bronchial asthma is a heterogeneous disease,and airway inflammation is one of the important mecha-nisms of its pathogenesis.Asthma can be classified into different types based on the different immune mechanisms involved in its pathogenesis,and the mechanism of airway inflammation also varies be-tween different types of asthma.This article reviews the research progress of miRNA in asthma airway inflammation and endotype,and explores the pathogenesis and treatment prospects of miRNA in asthma airway inflammation and endotype.
3.Microdissected peroneal artery perforator flap for repair soft tissue defect of dorsal fingers
Shaoxiao YU ; Wanggao ZHOU ; Guorong CHEN ; Zhenwei ZHANG ; Wenyi WU ; Xuelang YE ; Jinhao ZENG ; Yuhai KE
Chinese Journal of Microsurgery 2022;45(6):617-621
Objective:To investigate the clinical effect of microdissected peroneal artery perforator flap in repair of soft tissue defect of dorsal side of the fingers.Methods:From August 2015 to July 2020, 19 patients with soft tissue defects on dorsal fingers were treated with microdissected peroneal artery perforator flap. The area of wound defect was 3.8 cm×1.5 cm-5.8 cm×3.0 cm, with exposure of phalanges and tendons. The size of flaps was 4.0 cm×1.8 cm-6.0 cm×3.3 cm. According to the size of soft tissue defects on the dorsal side of the fingers, the flaps were designed with the perforating branch of peroneal artery in the centre. The length and width of a flap were 0.2-0.3 cm bigger and wider than the area of defect. The perforator vessels with a length of 2.0-3.0 cm were arvested in the superficial layer of deep fascia. Most of the adipose tissues of the flap were removed under microscope, and the small arteries between adipose tissues were protected. The flaps were used to cover the defects of fingers. The perforator artery of the flap was anastomosed with the proper palmar digital artery of the recipient site, the accompanying vein of the perforator artery was anastomosed with the dorsal digital vein of the recipient site, and the cutaneous nerve in the flap was anastomosed with the dorsal digital nerve. The donor sites were directly pulled together and sutured intermittently. Outpatient and WeChat follow-up were conducted after operation, including wound healing, flap survival, flap sensation, donor site recovery, and flexion and extension functions of the fingers. Functional recovery was evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Results:All wounds healed in Ⅰ stage, and all 19 flaps survived. The follow-up ranged from 9 to 25 months, with an average of 11.5 months. The appearance of the flaps was satisfactory and the texture was good. Sensation recoveried to S 4 in 4 paitients, S 3 in 9 patients and S 2 in 6 patients, and with only a linear scar was left in the donor sites. The hand function recovery was evaluated according to the Trial Criteria of Upper Limb Function Evaluation of the Hand Surgery Society of the Chinese Medical Association, with 18 cases were excellent and 1 was good. Conclusion:The microdissected peroneal artery perforator flap is an ideal surgical method to repair the soft tissue defect of dorsal side of the fingers, which has good shape and simple operation, avoids the secondary thinning and plastic surgery and offers good therapeutic effects.
5.Diagnostic value of serum antibody test and gastroscopy-guided duodenal drainage for clonorchiosis sinensis
Qihong YANG ; Chunling XU ; Yuhua CHEN ; Yongyi TAN ; Haizhen ZHONG ; Guorong YE ; Sujun HUANG ; Yuanran CHEN ; Xuyou LIU
Chinese Journal of Digestive Endoscopy 2021;38(8):638-643
Objective:To explore a fast method to identify and confirm suspected clonorchis sinensis infection.Methods:For suspected clonorchis sinensis infection, the clonorchiasis serum antibody was detected first with ELISA. If the antibody was positive, the fecal examination for eggs was performed. If the fecal examination was negative, duodenal drainage under gastroscopy was recommended to detect eggs from the drainage fluid.Results:A total of 126 patients met the requirements and aged 54.14±13.33 (24- 87). There were 83 cases (65.87%, 83/126) with eggs positive in the drainage fluid, of which 53 cases were male, aged 55.91±11.47 (30-86), and 30 cases female, aged 55.87± 13.85(30-87). There was no significant difference in age between males and females( P>0.05). The time of catheterization (T1) of 126 cases was 3.79 ±1.45 min. The time of drainage (T2) of 126 cases was 31.39 ±14.29 min. There was no significant difference in T1 or T2 between the positive group and the negative group( P>0.05). The detection rates of eggs were 91.57% (76 cases) in intrahepatic bile duct drainage, 81.93% (68 cases) in the bile-cyst juice and 75.90% (63 cases) in the common bile duct fluid. No serious adverse reactions occurred during or after the operation. Conclusion:The detection rate of clonorchiosis sinensis can be effectively improved by the combination of clonorchiasis serum antibody test and gastroscopy-guided duodenal drainage.
6.Application of 188 Re-labeled BaGdF5-PEG nanoparticles on the growth inhibition of hepatoma cells and the SPECT imaging in rabbit models
Tao WANG ; Ye PENG ; Xiao LI ; Guorong JIA ; Qiuhu WANG ; Chao CHENG ; Gaofeng SUN ; Changjing ZUO
Chinese Journal of Nuclear Medicine and Molecular Imaging 2018;38(11):721-725
Objective To investigate the inhibitory effect of 188 Re-labeled BaGdF5-poly ( ethylene glycol) ( PEG) nanoparticles ( NPs) on hepatoma cells, and explore the application of the radiolabeled NPs for SPECT imaging. Methods BaGdF5-PEG NPs were synthesized by hydrothermal method, and were fur-ther radiolabeled with 188Re using diethylene triamine pentaacetic acid (DTPA) as a coupling agent. The human hepatoma cells SMCC 7721 were treated with different concentrations of BaGdF5-PEG NPs, 188 ReO-4 or 188Re-DTPA-BaGdF5 NPs (14.8, 74.0, 370.0×104 Bq/ml) for 24 h, and then the cell proliferation rates were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. 188ReO-4 and 188 Re-DTPA-BaGdF5 NPs were administrated into normal rabbits via the ear vein, respectively. For the former, static SPECT/CT imaging were performed at 30, 60 min post-injection, and for the latter, dynamic SPECT images were captured within 10 min, and static SPECT/CT images at 30, 60, 120 min post-injec-tion. The rabbit VX2 tumor model was established, and a microcatheter was inserted into hepatic artery via the rabbit femoral artery, and then the mixture of 188 Re-DTPA-BaGdF5 NPs and lipiodol was injected into the tumor region. SPECT/CT imaging for VX2 tumor was performed at 30 min later. Data were analyzed by two-sample t test. Results The BaGdF5-PEG NPs were nearly square and the particle size was about 10 nm. The labeling yield of 188 Re-DTPA-BaGdF5 was 94.1% at the optimum conditions. Moreover, it showed high stability in vitro and in vivo. In vitro, BaGdF5-PEG NPs did not exhibit obvious cytotoxicity even at a high concentration. Both 188 ReO-4 and 188 Re-DTPA-BaGdF5 could inhibit the proliferation of SMCC 7721 cells, but 188 Re-DTPA-BaGdF5 showed a significantly stronger inhibitory effect at the doses of 74.0 and 370.0×104 Bq/ml ( t values:4.21,4.09, both P<0.01) . In vivo, 188 ReO-4 was absorbed by maxillary glands and was quickly elimi-nated from blood via the kidneys. The 188 Re-DTPA-BaGdF5 NPs mainly accumulated in the liver and spleen. In addition, retention and accumulation of 188 Re-DTPA-BaGdF5 NPs in the liver tumor could be achieved by using transarterial intervention technique for drug delivery. Conclusion 188Re-DTPA-BaGdF5 NPs have cer-tain killing effects on hepatoma cells in vitro, and with the help of transarterial intervention technique, the NPs can be aggregated within liver tumor, where they not only can be used for SPECT imaging, but also have potential therapeutic effects.
7.The effects of brain ischemic preconditioning on peripheral blood endothelial progenitor cells and neovas- cularization in rats
Xudong ZHANG ; Ye WANG ; Ran LIU ; Na WANG ; Guorong BI
Chinese Journal of Nervous and Mental Diseases 2015;(1):10-14
Objective To investigate the effects of brain ischemic preconditioning (BIP) on peripheral blood EPCs and neovascularization in ischemic brain tissue in rats with cerebral ischemia reperfusion injury (IRI). Methods One hundred and eight male SD rats were randomly divided into three groups:SO group (n=36), MCAO group (n=36) and BIP group (n=36). Neurological function assessment was conducted at 0 h before MCAO-reperfusion, 3 h, 24 h and 3 d, 5 d as well as 7 d after MCAO-reperfusion (n=6 for each group in each time point). Flow cytometry was used to calculate the number of EPCs. Immunohistochemical staining was used to detect the capillary density. Results ①Although neurologi?cal deficit scores were significantly decreased in both BIP and MCAO groups after 3 h following MCAO-reperfusion, the scores were much lower in BIP group than in MCAO group(5 d:1.00±0.63;7 d:1.00±0.63, P<0.05).②The numbers of EPCs were decreased in MCAO group while was increased in BIP group at 3 h after MCAO-reperfusion. The numbers of EPCs were significantly higher in BIP group than in MCAO group(24 h:0.58±0.07;3 d:0.80±0.10;5 d:0.68±0.05;7 d:0.52 ± 0.03, P<0.01). ③ The new blood vessels could be detected at 3 d in BIP group and 5 d in MCAO group after MCAO-reperfusion. The numbers of new blood vessels were significantly higher in BIP group than MCAO group(5 d:14.53 ± 3.44; 7 d: 41.40 ± 5.62, P<0.01). ④ Pearson analysis showed a positive correlation between EPCs and capillary density (5 d: r=0.855, P<0.01; 7 d: r=0.946, P<0.01). Conclusion BIP can improve EPCs mobilization and function, which may contribute to neovascularization in the ischemic brain tissue.
8.Research progress in antidotes of nerve agents in the USA
Jin CHENG ; Guorong DAN ; Yuanpeng ZHAO ; Jian WANG ; Feng YE ; Jiqing ZHAO ; Zhongmin ZOU
Military Medical Sciences 2015;(7):565-568
Nerve agent not only inhibit acetylcholinesterase ( AChE) at an early stage, but also induce prolonged and progressive neuroinflammation and delayed neurodegeneration.Recently, the US National Institute of Health ( NIH) has sponsored some major programs of toxic mechanisms and treatment of nerve agents, which aims at the development of quick and effective treatment to acute intoxication and delayed effect.The experimentally effective new antidotes mainly include AChE-targeting drugs, broad-spectrum reactivators and scavengers, antiinflamatory and nerve protection drugs.
9.Clinical Observation of Calcitriol Soft Capsules Combined with Telmisartan Tablets in the Treatment of Early Diabetic Nephropathy
Jianqiang YE ; Shunbin LI ; Guorong ZHANG
China Pharmacy 2015;(18):2470-2472
OBJECTIVE:To observe the clinical efficacy and safety of Calcitriol soft capsules combined with Telmisartan tab-lets in the treatment of early diabetic nephropathy(DN),and the effect on the levels of inflammatory factors. METHODS:Totally 110 patients with early DN were randomly divided into observation group and control group. The control group was orally given Telmisartan tablets with the initial dose of 40 mg,qd,and the maximum dose was 80 mg,qd;the observation group was orally giv-en Calcitriol soft capsules 0.25μg based on the treatment of control group,qd. The course was 1 month. The clinical data was com-pared,including the clinical efficacy and 24 h urinary protein,serum creatinine(Scr),urinary albumin excretion rate(UAER),se-rum C reactive protein (CRP),tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) before and after treatment. The adverse reactions were observed. RESULTS:After treatment,the total effective rate in observation group was significantly higher than con-trol group,with significant difference (P<0.05);the 24 h urinary protein,Scr,UAER,and levels of CRP,TNF-α and IL-6 in observation group were significantly lower than control group and before treatment,with significant differences(P<0.01 or P<0.05). There were no obvious adverse reactions in 2 groups. CONCLUSIONS:Calcitriol soft capsules combined with Telmisartan tablets has better efficacy than only Telmisartan tablets in the treatment of DN,and can more effectively improve the levels of CR, TNF-αand IL-6,which is helpful to delay progression of patients.
10.Effect of glucose concentrations on energy metabolism in 16 HBE cells exposed to CEES
Feng YE ; Guorong DAN ; Jian WANG ; Yuanpeng ZHAO ; Jiqing ZHAO ; Zhongmin ZOU
Military Medical Sciences 2014;(3):169-173
Objective To compare the changes in energy metabolism in 2-chloroethyl ethryl sulfide(CEES)-poisoned bronchial epithelial cell 16HBE cultured in media at different glucose concentrations .Methods Bronchial epithelial cell 16HBE was cultured in high (4.5 mg/ml) or low (1.1 mg/ml) glucose medium and exposed to a sulfur mustard simulant CEES of 0.2, 0.5, 1.0 mmol/L.Cell growth and cytotoxicity were tested using MTS .ATP, ADP and AMP were detected by HPLC and the value of ATP/ADP, total adenine nucleotides ( TAN) and energy charge ( EC) was subsequently calculat-ed.Mitochondrial oxidative phosphorylation-related proteins, COX-10 and ISCU, were detected using Western blotting . Rhodamine 123 was applied to detect the mitochondrial membrane potential using flow cytometry .Results Low glucose accelerated the growth and energy metabolism of 16HBE cells in regular culture , and the contens of ADP , TAN, COX-10 and ISCU in low glucose group were significantly higher than those in high glucose group .CEES exposure (≥0.5 mmol/L) significantly affected cell viability in both high and low glucose groups , with significant difference between the two groups exposed to 1.0 mmol/L CEES.In high glucose group, 24 h after 0.5 or 1.0 mmol/L CEES exposure, the contents of ATP, ADP and TAN were significantly increased , while ATP/ADP and EC decreased .In low glucose group , ADP, AMP and TAN significantly decreased, while ATP/ADP and EC increased 24 h after 1.0 mmol/L CEES exposure.The mi-tochondrial membrane potential (MMP) also changed differently after 0.5 mmol/L CEES exposure.MMP in high glucose group marginally increased at 3 h, and significantly increased at 8-12 h (P<0.05), and returned to normal at 24 h. MMP in low glucose group showed a transient decrease at 5 h (P<0.01), and back to normal at 8 h.The protein levels of COX-10 and ISCU were significantly increased in high glucose group 24 h after 0.5-1.0 mmol/L CEES exposure , but sig-nificantly decreased in low one 24 h after 1.0 mmol/L CEES exposure .Conclusion When 16HBE is cultured at a high or low glucose concentration , the cell growth, stress responses and energy metabolism including MMP , COX-10, ISCU and ATP production are in different status before or after CEES exposure .High glucose could protect against CEES exposure .

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