1.Progress in mechanism and detection methods of ferrinophagy-mediated ferroptosis
Jie GUO ; Yulong WANG ; Fengyi MAI ; Wentao YANG ; Jingrong LIANG ; Junxiang SHU ; Chenguang LI
Chinese Journal of Pathophysiology 2024;40(2):365-374
Ferroptosis is an iron-dependent form of regulated cell death,which is distinct from apoptosis,ne-crosis,and pyroptosis.Recent studies have found that activators of ferroptosis,such as Erastin,can activate autophagy-re-lated proteins,induce the formation of autophagosomes,and ultimately release ferric ions to mediate ferroptosis.This pro-cess,called ferritinophagy,is initiated by the binding of an autophagic cargo receptor protein,nuclear receptor coactivator 4(NCOA4),to iron-laden ferritin.The transfer of NCOA4-ferritin to the lysosome by ferritinophagy results in the proteoly-sis of ferritin,and,in turn,the release of its iron content and lipid-reactive oxygen species(ROS)accumulation.Ferritin-ophagy has been closely associated with central nervous system disorders,circulatory system diseases,and cancer.Fur-thermore,the regulation mechanism of ferritinophagy is also a hot topic in the study of iron-dependent cell death process.With the in-depth study of ferritinophagy,great progress has been made in the study of key components of ferritinophagy as well as its molecular mechanisms and processes.However,a comprehensive summary of the methods for detecting ferritin-ophagy is still unclear.To further deepen the understanding of ferritinophagy and its detection methods,this review focus-es on the concept,characteristics,methods,and precautions during detection of ferritinophagy.This review provided ex-perimental reference for subsequent researchers and promoting the progress of research related to ferritinophagy.
2.Establishment of depression model in 3xTg-AD transgenic mice and comparison of cognitive-related pathological changes
Jingrong TANG ; Xiaoyu YU ; Quanxian GUO ; Yanuo WEI ; Jiaxin TIAN ; Shengxi WU ; Jie XIANG
Chinese Journal of Neuroanatomy 2024;40(2):162-170
Objective:To analyze the effect of depressive state on Alzheimer's disease(AD)mice,3xTg-AD mice were stimulated with chronic social frustration stress(CSDS)and chronic mild unpredictable stress(CUMS),to estab-lish an early AD-induced depression mouse model,and to detect cognitive and behavioral changes,activation of micro-glia in the hippocampus and neuronal loss.Methods:Three-month-old mice were subjected to 8-day stress stimulation alternately,the depressive state of the mice was evaluated by behavior,the evaluation criteria were formulated,and the cognitive behavior was detected and analyzed,and the hippocampal brain tissue sections were stained with immunofluo-rescence to observe the deposition of β-amyloid(Aβ)and the aggregation of microtubule-associated protein(tau),mi-croglia activation and neuronal loss.Results:Depression-related behavioral results showed that the CSDS+CUMS group had depression-related phenotypes.Cognitive-behavioral testing showed that the new object recognition index of the mice in the CSDS+CUMS group was significantly reduced(P<0.05),and the Morris water maze showed that the spatial memory ability of the CSDS+CUMS group was significantly reduced(P<0.05),but there was no obvious fear memory loss in the CSDS+CUMS group in the conditioned fear experiment.The results of immunofluorescence staining showed that Aβ deposition appeared in the hippocampus at 4 months of age,the activated microglia increased(P<0.001),and a certain degree of neuronal loss appeared in the CSDS+CUMS group(P<0.001);At 8 months of age,the CSDS+CUMS group showed tau protein aggregation early.Conclusion:We established a model of AD-induced de-pression in AD mice,in which 3xTg-AD mice experienced early decline in learning memory and increased AD-related pathological deposition of neurons in the hippocampus,accompanied by microglial activation and neuronal loss.
3.Development and evaluation of portable pulse vacuum pressure steam sterilizer
Yuequan LUO ; Jingrong WEI ; Bin LI ; Yang LUO ; Jiwei GUO
Chinese Journal of Nursing 2017;52(4):489-492
Objective Due to poor effects of sterilization,low efficiency and inconvenience of field emergency transportation for portable exhaust pressure steam sterilizer,we aimed to design and develop a portable pulse vacuum pressure steam sterilizer in order to effectively improve quality and efficiency of sterilization for sterile medical equipment in disaster and emergency medical treatment.Methods From the beginning of February 2013,we focused on the features of small size,light weight,reliable sterilizing effect and fast speed of sterilization,and conducted series of technology researches,and finally developed portable pulse vacuum pressure steam sterilizer which was suitable for individual carrying or air delivery.Results We performed outdoor sterilization tests for portable vacuum pressure steam sterilizer and portable exhaust pressure steam sterilizer under the same condition.Portable pulse vacuum pressure steam sterilizer achieved a 100% qualification rate,average sterilization cycle was 28.6min per time,sterilization by weight was 48L/h,average water consumption was 0.4L/boiler,average power consumption was 0.7kw/h per time,and sterilization operation index,maneuverability and transportability were significantly better than those of portable exhaust pressure steam sterilizer.Conclusion The developed 4 portable pulse vacuum pressure steam sterilizer have higher cost performance,and have been successfully applied for medical service training and non-war medical treatment activities in our hospital.
4.Effect of Ai'er health exercise in hospitalized patients with chronic heart failure
Jingrong HAO ; Li GUO ; Changming REN ; Chi ZHANG
Chinese Journal of Modern Nursing 2017;23(11):1518-1521
Objective To explore the effect of Ai'er health exercise in patients with chronic heart failure.Methods A total of 110 hospitalized patients with chronic heart failure were randomly divided into Ai'er health exercise group and control group, with 55 cases in each group. The patients of control group were given routine treatment and nursing, patients in Ai'er health exercise group were given Ai'er health exercise based on the control group treatment and nursing method. The cardiac function classification, cardiac function index, 6 min walking test, 6 months after discharge and the rate of hospitalization and mortality were compared between two groups before and after the intervention.Results After the intervention, heart function index of the two groups of patients were significantly improved, and the differences were statistically significant (P<0.05). The degree of the heart function index of the health of Ai'er exercise group was significantly better than that of the control group, and the difference was statistically significant (P<0.05). After the intervention, the health of Ai'er exercise group for 6 min walking test distance were (220.8±13.5)m which was significantly longer than that in the control group (153.4±9.7)m, and the difference was statistically significant(t=3.139,P<0.05). Ai'er health exercise patients were discharged after 6 months of hospitalization was 7.3% which was lower than that in the control group 21.8%, and the difference was statistically significant (P<0.05); the mortality rate of two patients discharged from 6 months had no significant difference (P>0.05).Conclusions Ai'er health exercise can significantly improve the heart function index in hospitalized patients with chronic heart failure, improve exercise capacity, and reduce readmission rate.
5.Effects of miR-145 on the proliferation, apoptosis and cell cycle of a human keratinocyte cell line HaCaT
Jingrong LI ; Jianqin WANG ; Ruihua FANG ; Renshan ZENG ; Jinxue MO ; Yunlong GUO ; Shuqing JIA
Chinese Journal of Dermatology 2015;48(6):416-420
Objective To investigate the regulatory effects of miR-145 on the proliferation,cell cycle and apoptosis of a human keratinocyte cell line HaCaT.Methods miR-145 mimics and negative control (NC) mimics were chemically synthesized and then transiently transfected into HaCaT cells respectively.After additional culture for different durations,real-time PCR was performed to determine the expression level of miR-145,MTS assay to estimate cell proliferation,and flow cytometry to detect cell apoptosis and cycle.Luciferase assay,real-time PCR and Western blot were conducted to determine whether NRAS was the target gene of miR-145.Results The miR-145 expression level in miR-145 mimic-transfected cells increased by 85.00 ± 1.21 folds compared with NC mimic-transfected cells (t =115.90,P < 0.0001).The transfection with miR-145 mimics significantly inhibited the proliferation of HaCaT cells (F =8.76,P =0.008),and the inhibitory effect significantly varied with the duration (24-96 hours) of culture after transfection,with no interaction effect between the transfection with miR-145 mimics and culture duation (F =1.21,P =0.18).Compared with NC mimic-transfected cells,those transfected with miR-145 mimics showed a significant increase in the proportion of early apoptotic cells (18.9% ± 4.1% vs.4.3% ± 1.2%,t =7.126,P < 0.01),late apoptotic cells (9.3% ± 2.3% vs.3.6% ± 1.6%,t =12.38,P < 0.01),G1-phase cells (85.83% ± 5.2% vs.62.08% ± 6.23%,t =11.78,P =0.007),but a significant decrease in the percentage of G2-phase cells (6.26% ± 1.2% vs.19.36% ± 3.45%,t =7.610,P =0.017) and S-phase cells (7.91% ± 1.3% vs.18.56% ± 5.23%,t =7.230,P=0.019).As luciferase assay showed,luciferase activity was significantly lower in HaCaT cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-wild carrying the wild-type 3'UTR of NRAS than in those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-wild (t =11.09,P =0.008),but similar between cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-mut carrying the mutant-type 3'UTR of NRAS and those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-mut (P > 0.05).Real-time PCR and Western blot revealed that the overexpression of miR-145 mimics had no significant effect on NRAS mRNA expression (P > 0.05),but significantly inhibited NRAS protein expression (1.52 ± 0.07 vs.0.20 ± 0.02,t =28.43,P< 0.01).Conclusion miR-145 might inhibit proliferation and promote apoptosis of HaCaT cells by influencing cell cycle via NRAS.
6.Research on PCR determining of type b Haemophilus influenzae strains and immunogenicity of polysaccharide conjugates
Yulan YUAN ; Jingrong GUO ; Jiwei ZHOU ; Hua GAO
Chinese Journal of Microbiology and Immunology 2012;32(4):346-350
Objective To determine Haemophilus influenzae type b strains in molecular level using PCR,and to study the immunogenicity of capsular polysaccharide conjugates in mice.Methods Extracting genomes using bacterial DNA extract kit from Hoemophilus influenzae type b strains,and PCR for determining the strains through serotyping-specific and capsular genotyping primers respectively.Various capsular polysaccharides conjugated TT respectively,and the conjugates were administered subcutaneously to mice through dilution.After vaccination with two doses,blood samples were collected for the detection of antibody levels to polyribosylribitol phosphate ( PRP),the capsular polysaccharide of Hib.Results All five Haemophilus influenzae type b strains contain type-specific(482 bp) and capsular type (343 bp)DNA fragment through PCR detecting.The DNA fragments were sequenced.BLAST show that these sequences are 100% homology comparing the above strains respectively,and are 99% and 100% homology comparing the GenBank X78559.1 and M19995.1 respectively.The immunogenicity of mice from various capsular polysaccharide conjugates (PRP-TT) was not significantly different by ELISA detecting.Conclusion Through PCR,Haemophilus influenzae type b strain can be determined in molecular level.The immunogenicity of mice from purified capsular polysaccharide conjugates was not different.The study provides a detection means for the features and heredity stability of Haemophilus influenzae type b strain and reference data for the immunogenicity of different polysaccharide conjugates in vaccine research and development and production.
7.Subtype and sequence analysis of gag genes in HIV-1 circulating in sexual infectors in Beijing
Jingrong YE ; Lei GUO ; Lishi BAI ; Ruolei XIN ; Hongyan LU ; Shuangqing YU ; Yi ZENG
Chinese Journal of Microbiology and Immunology 2011;31(2):136-139
Objective To investigate the subtype distribution and sequence characteristics of HIV-1 strains prevalent among sexual infectors in Beijing. Methods We collected the blood samples from 100HIV sexual infectors in Beijing during 2008 and separated plasma specimens. RNA was extracted from the plasma and the gag gene was amplified by RT-PCR and nest-PCR. The PCR products were sequenced directly and phylogenetic analyses of gag gene was performed using the MEGA4 software. Results Among 100 HIV-1 plasma samples,84 gag gene fragments were amplified and analyzed. Eight HIV subtypes including B(22 strains), B'(8 strains),C( 1 strain) ,CRF01_AE (38 strains) ,CRF02_AG (2 strains) ,CRF07_BC(9 strains) ,CRF08_BC(3 strains) and C/CRF01_AE recombinant like strain( 1 strain) were identified circulating in Beijing. Conclusion CRF01 _AE and subtype B were predominant in Beijing account for 45.2% and 26.2% and the surveillance of HIV gene variation should be paid more attention.
8.Expressions of survivin, COX-2, VEGF and their correlations with angiogenesis in condyloma acuminatum
Liehua DENG ; Jingrong LI ; Yunfeng HU ; Xiangqian XU ; Yan WU ; Jin XU ; Xiuzhi GUO ; Jie LIU ; Hongtao FAN ; Ze LIN ; Yongkeng ZHAO
Chinese Journal of Dermatology 2010;43(11):766-769
Objective To investigate the expressions of survivin, cyclooxyenase-2 (COX-2) and vascular endothelial growth factor (VEGF) and their relationship with angiogenesis in condyloma acuminatum (CA) tissues. Methods Immunohistochemistry using PowerVision staining kit was performed to detect the expression of survivin, COX-2 and VEGF protein in 60 CA tissue samples from patients and 21 normal skin samples from the foreskin of human controls. At the same time, the microvessel density was determined in CA tissues by staining blood vessel endothelium with anti-CD105 monoclonal antibody. Results The positivity rate of survivin and COX-2 expression was 56.67% and 63.33%, in CA tissues, 9.52% and 0 in normal skin tissues, respectively. There was a significant difference between the two groups of tissue samples in the positivity rate and intensity of survivin and COX-2 expression (all P < 0.05). VEGF was expressed in all of the CA tissues and normal skin tissues, while the intensity of VEGF expression was statistically different between the two groups of tissue samples (P < 0.05). The MVD was 16.38 ± 5.46 and 0.62 ± 0.44 in CA tissues and normal skin tissues, respectively (P < 0.05). There was a significant positive correlation between the expressions of survivin, COX-2 and VEGF, as well as between MVD and the expressions of survivin and COX-2 in CA tissues. Conclusion The expression levels of survivin, COX-2 and VEGF are significantly higher in CA tissues than in normal skin tissues.
9.Study on the immunogenicity of group A plus group C meningococcal glycoconjugate vaccine
Jianhua WANG ; Meiying LIU ; Yeshan LI ; Jingrong GUO ; Yanbin ZHANG ; Yun CHEN ; Hua GAO ; Yun LIN ; Yu LIU
Chinese Journal of Microbiology and Immunology 2008;28(2):127-129
Objective To investigate the immunogenicity of group A plus group C meningococeal glycoconjugate vaccine,namely dosage,immune memory and compatibility. Methods The mice were injected with group A plus group C meningococcal glycoconjugate vaccine with different dosages. Blood samples were taken on the 14th day after the last injection for testing the antibodies against polysaccharide A and C. After the optimal immunization dosage had been decided,the mice were inoculated separately with the monovalent group A and the monovalent group C and the bivalent group A plus group C glycoconjugate vaccine with one,two or three injections for observation of the effectiveness of different injections and the compatibilities. Results The dosage of 1.25 μg of each polysaccharide of group A and group C in bivalent glycoconjugate vaccine appears to be immunologically optimal to vaccinate the mice. Immunological memory could be induced in mice inoculated with the glycoconjugate vaccine,and the antigenic immunogenicity of the group A component and group C component in the formulation of group A plus group C meningococcal glycoconjugate vaccine was not affected. Conclusion Group A plus group C meningococcal glycoconjugate vaccine have good immunogenicity,immune memory and compatibility.
10.Impact of Expression of DHA AmpC ?-Lactamases Mediated by Plasmid on Antibiotics Susceptibility of Clinical Isolates of Klebsiella pneumoniae
Yanping LUO ; Dingxia SHEN ; Zheng CUI ; Jingrong CAO ; Yaping XU ; Junjie GUO ; Rong CHEN
Chinese Journal of Nosocomiology 2006;0(05):-
OBJECTIVE To investigate the prevalence of DHA AmpC ?-lactamases mediated by plasmid in Klebsiella pneumoniae in China.METHODS Antimicrobial susceptibility test was conducted by the methods of double agar dilution and ESBLs confirmatory in K-B method according to the criteria of guidelines of CLSI.AmpC ?-lactamases were detected on the basis that AmpC ?-lactamases could be inhibited by 3-aminophenylboronic acid(APB).Gene chip technology and PCR were used to detect ESBLs and AmpC gene.RESULTS Among total 34 isolates of K.pneumoniae 32(94.1%) produced AmpC ?-lactamases and ESBLs.The most common(38.3%) were types DHA and TEM and SHV.MIC50 and MIC90 of all strains to all tested antimicrobial agents were lower than 34 strains tested 0.25?g/ml and 0.5?g/ml.Fourteen strains AmpC and ESBLs were conjugated successfully.CONCLUSIONS DHA AmpC ?-lactamases mediated by plasmid are the most common in K.pneumoniae in General Hospital of PLA of China.The most common(38.3%) are types DHA and TEM and SHV.Fourteen(41.2%) strains can be spreaded by plasmid.

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