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.Re-detectable positive SARS-CoV-2 RNA tests in patients who recovered from COVID-19 with intestinal infection.
Wanyin TAO ; Xiaofang WANG ; Guorong ZHANG ; Meng GUO ; Huan MA ; Dan ZHAO ; Yong SUN ; Jun HE ; Lianxin LIU ; Kaiguang ZHANG ; Yucai WANG ; Jianping WENG ; Xiaoling MA ; Tengchuan JIN ; Shu ZHU
Protein & Cell 2021;12(3):230-235
3.Application of progressive nutrition guide sheet in postoperative enteral nutrition for patients with oral cancer
Tian ZHANG ; Dan QIAO ; Yan ZHANG ; Yuan TANG ; Xiongtao YANG ; Jie HE ; Xianhong LAI ; Liping XIAO ; Dan LI ; Guorong WANG ; Tao YU
Chinese Journal of Clinical Nutrition 2021;29(1):22-29
Objective:To explore the effect of applying individualized progressive nutrition guide sheet in postoperative enteral nutrition (EN) for patients with oral cancer.Methods:Using convenient sampling method, 40 oral cancer patients admitted to Sichuan Cancer Hospital from November 2017 to October 2018 were selected as the control group, and 46 from November 2018 to October 2019 were selected as the observation group. Both groups received EN support but the observation group were applied with progressive nutrition guide sheet. The pre- and post-operative body weight, nutrition related indicators, gastrointestinal symptoms, proportion of patients achieving daily target energy intake, patient/family satisfaction and other indicators were compared between the two groups.Results:There were significant differences in preoperative potassium, total protein and albumin at 7 days after operation, prealbumin at 3 and 7 days after operation, potassium at 3 days after operation and sodium at 3 days after operation between the two groups( Z=4.963, P<0.01; Z=5.094, P<0.01; Z=-2.022, P<0.05; Z=4.048, P<0.01; Z=2.14, P<0.05, Z=-6.04, P<0.01, Z=-7.13, P<0.01). The dynamic changes of potassium and sodium in the two groups were compared before operation, 3 days after operation and 7 days after operation ( F=30.20, F= 118.51, all P<0.01). There were significant differences in incidence of abdominal pain, abdominal distension and diarrhea between the two groups ( χ2=6.91, P=0.009, χ2=10.36, P=0.001, χ2=4.71, P=0.03). There were also significant differences in the proportion of patients achieving daily target energy intake at 1 day, 2 days, 3 days, 4 days, 5 days, and 6 days after operation between the two groups ( χ2=41.77, χ2=45.09, χ2=45.71, χ2=40.53, χ2=29.97, χ2=6.11, all P<0.01). Conclusion:The application of progressive nutrition guidelines in early postoperative EN support for patients with oral cancer can help to improve postoperative nutritional status, avoid potassium, sodium and electrolyte disturbance, alleviate postoperative gastrointestinal symptoms, improve the achievement of daily target energy intake and patient/family satisfaction, and promote disease recovery.
4.Strategies and progresses of stereotactic radiosurgery for intractable brain metastases
Chinese Journal of Radiological Medicine and Protection 2020;40(5):408-412
Brain metastases are common intracranial diseases featured with high incidence rate, difficult clinical treatment and poor prognosis. Especially for refractory brain metastases (large volume, multiple recurrence and metastatic tumors), it is difficult for conventional therapies to achieve ideal prognosis. Compared with conventional therapies, stereotactic radiosurgery (SRS) has the advantages of high accuracy, large fractional dose and low damage to the surrounding tissues, and has gradually become the primary choice for the treatment of brain metastases. SRS has a good effect in the treatment of refractory brain metastases, and can effectively inhibit tumor proliferation and improve the quality of life of patients. SRS combined with surgery, chemotherapy, molecular targeted therapy and immunotherapy can improve the local tumor control rate of patients with intractable brain metastasis, but whether it can improve the prognosis of patients is still controversial. The adverse reactions of combined therapies should also be concerned. Therefore, this article reviews the therapeutic effects of SRS in the treatment of refractory brain metastases, the strategies and progresses of combined treatment.
5.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.
6.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 .
7.miR-34a partially reverses inhibition of CEES-exposed keratinocytes migration via ERK1/2 pathway
Feng YE ; Jian WANG ; Guorong DAN ; Tao SHANGGUAN ; Jiqing ZHAO ; Yuanpeng ZHAO ; Zhongmin ZOU
Military Medical Sciences 2014;(11):845-849
Objective To explore the effect of 2-chloroethyl ethyl sulfide(CEES) poisoning on keratinocyte migration and the regulatory role of microRNA(miR)-34a.Methods MTS was used to detect the viability of cells exposed to CEES in order to select an appropriate dose of CEES exposure in this in vitro model.The protein level of keratin 5 and keratin 10 was detected to assess cell differentiation status .Scratch assay was applied to evaluate cell migration ,and miR-34a silencing in keratinocytes was achieved by transfecting chemically synthesized miR-34a specific miRNA inhibitor.t-ERK1/2 and p-ERK1/2 levels closely related to cell migration were detected using Western blotting .Results An in vitro CEES exposure model of keratinocytes was established at the optimal concentration of 0.5 mmol/L CEES in the viability test , and this dose was chosen to evaluate cell migration changes .The migration of cells was significantly inhibited 24 h after CEES exposure , accompanied by no changes in morphology and keratin 5/10 levels.Silencing of miR-34a significantly increased the migration of cells exposed to CEES , which could be blocked by adding 5 μmol/L U0126 , an ERK1/2 phosphorylation selective inhibitor.Conclusion Silencing of miR-34a can significantly increase keratinocyte migration and partially reverse the inhibition of CEES-caused migration , which could be mediated by ERK 1/2 pathway activation .
8.Analysis of epidermal growth factor receptor expression and gene expression status in tissue microarray of cervical squamous cell carcinoma
Suying ZHOU ; Guofei FENG ; Guorong CHEN ; Dan PAN ; Pinnan ZHANG ; Xiaomin YANG ; Zuoli XIA
Chinese Journal of Obstetrics and Gynecology 2013;48(11):843-846
Objective To elucidate the protein expression and gene expression status and the relationship between epidermal growth factor receptor (EGFR) protein expression and EGFR gene status.Methods Tissue microarray containing 72 cervical squamous cell carcinoma tissues was constructed,and EGFR protein expression and gene status were evaluated by immunohistochemical and fluorescence in situ hybridization (FISH) techniques.Results Protein expression of EGFR:69 of 72 cervical squamous cell carcinomas were observed.The results demonstrated it was significant association with invasion depth,lymphnode metastasis and lymph-vessel invasion (x2 =4.998,P < 0.05 ; x2 =4.299,P < 0.05 ; x2 =4.686,P < 0.05) in cervical squamous cell carcinomas.For FISH assessing EGFR gene,64 of 72 carcinomas were observed; 7 of 64 cases showed EGFR gene amplification,and 25 disomy,23 trisomy and 9 polysomy were detected.There were high levels of protein expression in all the EGFR gene amplification cases,and there were significant association between EGFR protein expression and the gene copy number (x2 =13.564,P < 0.05).Conclusions EGFR may participate in the occurrence,progression and metastasis of cervical squamous cell carcinoma.Overexpression of EGFR protein may result from gene amplification and gene copy number increases,which showed that EGFR gene expression status may be a more effective biological indicator of cervical squamous cell carcinoma targeted therapy.
9.Effects of hypoxia and sodium cyanide on superoxide dismutase,reduced glutathione and malondialdehyde in arterial blood of rabbits
Yuanpeng ZHAO ; Guorong DAN ; Zhaojun DONG
Journal of Third Military Medical University 2003;0(16):-
Objective To investigate the effects of hypoxia and sodium cyanide(NaCN)on oxidative stress in rabbit arterial blood.Methods An artificial hypobaric hypoxia chamber was used to simulate 4 000-meter high altitude.Twenty rabbits were randomly divided into 4 groups:hypoxia group with high-or low-dose,non-hypoxia with high-or low-dose.The animals in the non-hypoxia groups were operated under normal circumstances while those in hypoxia groups were subjected to chamber in low pressure for 72 h before receiving the hypoxia experiments.Femoral arterial cannulation was performed on all animals under anesthetization with pentobarbital sodium(30 mg/kg,iv)and NaCN(ip)at the doses of 1.5 mg/kg and 2 mg/kg.Blood samples were collected at 10 min before intoxication,and 5,10,15,20,30,60,120 and 180 min after intoxication and blood seperation was conducted.The activity of superoxide dismutase(SOD),contents of reduced glutathione(GSH)and malondialdehyde(MDA)were determined.Results The MDA content was significantly increased(P
10.Numbers and Species of Microorganisms of 5 Probiotic Products for Medicinal Purposes
Guorong DAN ; Jing YUAN ; Huan TANG ; Hong WEI
China Pharmacy 2001;0(07):-
OBJECTIVE:To compare whether the numbers and species of strains of5probiotic products for medical pur-poses sold in the market are in line with those stated on the labels.METHODS:Viable count of probiotic products was per-formed by selective plating count method and species of the isolated strains were analyzed and identified using16S rDNA se-quencing.RESULTS:Regarding viable count,only one of the total25batches of samples in5products was lower than that stated on the label.As for species,of the10isolated strains,3failed to be in consistent with that stated on the label.CONCLU_ SIONS:For the5products investigated,inconformity was noted between strain species and that stated on the labels,however,the same viable counts are noted between the products and that stated on the labels.

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