1.Expression of GRP78 and CHOP in lungs of rats with bronchopulmonary dysplasia and their relationship with pneumonocyte apoptosis
Chuchu GAO ; Hongyan LU ; Wei TANG
Journal of Clinical Pediatrics 2013;(10):959-963
Objectives To investigate the expressions of endoplasmic reticulum stress (ERS)-related factors, glucose regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP), in lungs of rats with bronchopulmonary dysplasia (BPD) and their relationship with pneumonocyte apoptosis, and further to explore the role of ERS-induced apoptosis in the pathogen-esis of BPD. Methods A total of 24 neonatal SD rats were randomly divided into control group and BPD group with 12 rats each. Rats in BPD group were exposed to 85%O2, while rats in the control group were exposed to air. Four rats in each group were sacriifced at 7, 14 and 21 days respectively after exposure. The expression of GRP78 in the lung tissues was examined by immunohistochemistry, the mRNA and protein levels of GRP78 and CHOP were detected respectively by real-time PCR and Western blot, and the apoptosis in lung cells were evaluated by TdT-mediated dUTP nick end labeling (TUNEL) assay. Results The mRNA and protein levels of GRP78 and CHOP, and the cell apoptosis in BPD group were signiifcantly different from those in control group (P<0.01) at different time points and increased over the time of hyperoxic exposure (P<0.01). Conclusions En-doplasmic reticulum stress may be initiated by hyperoxic exposure and apoptosis is induced via CHOP signal pathway, which is involved in the pathogenesis of BPD.
2.Cloning and prokaryotic expression of rat surfactant protein C in E.coli
Wei TANG ; Hongyan LU ; Chuchu GAO
Journal of Clinical Pediatrics 2014;(2):183-185
Objectives To construct prokaryotic expression plasmid of rat surfactant protein C (sp-c) gene under hyperoxia and expression in E.coli. Methods Twenty-one-day-old SD premature rats were exposed to 85%hyperoxia 12 hours after birth. The rats were executed after 7 days and their RNA were extracted from lung and cDNA was synthesized and amplified. And then the cDNA was cloned into pMD18-T vector and confirmed by enzyme digestion and sequencing. After the prokaryotic expression vector pET-28a(+)-sp-c was constructed, the recombinant plasmid was induced by IPTG and expressed in E.coli BL21 strain. The fusion protein was analyzed by SDS-PAGE and Western blotting. Results The pET-28a(+)-sp-c plasmid was constructed and the fusion protein with relative molecule mass of 21000 was highly expressed. Conclusions SP-C is successfully expressed in E. coli, which can be used to study the mechanism of action between SP-C and endoplasmic reticulum, the expression of SP-C in lung epi-thelial cell A549 and the impact of SP-C on proliferation, differentiation and apoptosis of AECIIs in future.
3.Research progress of smartphone addiction interventions in adolescents
XU Chuchu, XIE Jin, WEI Xinyu, HUANG Hai, ZHOU Chunyan
Chinese Journal of School Health 2022;43(8):1276-1280
Abstract
Adolescents at the unique period during which individuals experience rapid growth in social, emotional, and cognitive aspects, are more prone to smartphone addiction than other groups. Smartphone addiction has been proved to seriously affect the physical and mental health of adolescents, and these impacts will become increasingly widespread with age. The effectiveness of current smartphone addiction interventions are mild among adolescents, which have proved to be related to adolescents unique cognitive and psychological characteristics. This paper reviews the latest research on adolescent smartphone addiction interventions, and classifies and elaborates from information enhancement strategies, ability enhancement strategies and behavioral intervention strategies, and puts forward the feasibility suggestions in this field, providing reference for localized intervention strategies, with the aim to developing systematic intervention design guidance and an effective multi subject collaborative intervention system.
4.Correlation analysis of immune antibodies with pelvic inflammatory diseases
Fang LIANG ; Hanlin XIE ; Yanxing LIU ; Peiqi WEI ; Zhenghe SHENG ; Yinghong WENG ; Jingchun QIN ; Jian ZENG ; Chuchu WEI ; Dan SONG ; Suzhang LIU ; Yuanyue ZHU ; Ziyu LYU
Immunological Journal 2024;40(5):480-484
This study was designed to evaluate the correlation between immune antibodies and pelvic inflammatory disease(PID)using retrospective analysis.Cases were selected from 171 patients who met the diagnosis of PID in Liuzhou People's Hospital of Guangxi Province from January 2022 to March 2023,and the PID patients were further divided into simple PID group(53 cases)and in PID combined with reproductive tract infection group(118 cases)according to the presence or absence of reproductive tract infections,while 83 cases of women who did not meet the specific diagnostic criteria of PID and did not have reproductive tract infections were selected as the control group during the same period.The positive rate of immune antibodies in the three groups were observed and compared to explore the relationship between immune antibodies and PID.Data showed that the positive rates of immune antibodies were significantly higher in the PID alone group and the PID combined with reproductive tract infection group than that in the control group.Furthermore,the positive rate of immune antibody TPOAb was significant difference in the PID combined with reproductive tract infection group and the PID alone group(P<0.05).In conclusion,TPOAb is closely associated with reproductive tract infections.
5.Gene mutation type and clinical phenotype of patients with PRRT2 mutation and their relations with prognosis
Yajing GAN ; Jiewen DENG ; Guoyan LI ; Zihan WEI ; Yan FENG ; Yuqing SHI ; Chuchu ZHANG ; Yanchun DENG
Chinese Journal of Neuromedicine 2024;23(9):895-902
Objective:To analyze the gene mutation type and clinical phenotype of patients with PRRT2 mutation, and explore their relations with prognosis. Methods:A total of 18 patients with PRRT2 gene mutation (1 patient with novel mutation in PRRT2 gene, and 17 probands in 17 families with PRRT2 gene mutation) were enrolled in Department of Neurology, First Affiliated Hospital of Air Force Medical University from January 2018 to July 2023. Serum of the patients was collected for whole exon sequencing, and mutation sites and types of PRRT2 gene were analyzed. SWISS-MODEL website was used to predict the changes in protein structure caused by PRRT2 gene mutation. The relations of gene mutation type and clinical phenotype with prognosis of these patients were analyzed. Results:(1) All 18 patients with PRRT2 gene mutation were heterozygous mutation, including 12 frameshift mutations, 5 missense mutations, and 1 integer mutation. The clinical phenotype included benign familial infantile epilepsy (BFIE) in 5 patients, epilepsy in 6 patients, exercise-induced paroxysmal kinesigenic dyskinesia (PKD) in 5 patients, and infantile convulsion and choreoathetosis (ICCA) in 2 patients. A total of 8 mutation sites were found in 18 patients with PRRT2 gene mutation, of which 3 mutation sites have been reported, and 5 mutation sites have not been reported, including c.647(exon2)C>A, c.647(exon2)C>G, c.170(exon2)delC, c.981(exon3)C>G, and lossl(EXON: 2)(all). (2) Eighteen patients mainly accepted oxcarbazepine, levetiracetam, and sodium valproate in combination or monotherapy. Among them, 5 BFIE patients, 2 ICCA patients and 3 epilepsy patients were seizure-free after treatment. PKD patients did not respond well to oxcarbazepine. (3) Three frameshift mutations (mutation sites: c.649 [exon2]_c.650 [exon2] insC, c.640 [exon2]_c.641 [exon2] insC, and c.170 [exon2] delC) led to premature termination of protein translation, resulting in significant changes in protein structure. Four missense mutations (mutation sites: c.640[exo2]G>C, c.647[exon2]C>A, c.647[exon2]C>G, and c.981[exon3]C>G) had little effect on protein structure changes. No relation was found between changes of protein structure caused by different mutation types and prognosis. Conclusion:PRRT2 gene mutation patients with clinical phenotypes of BFIE and ICCA have good prognosis, but the mutation type is not related with the prognosis of patients.
6.Tracing and transmission analysis of a nosocomial infection event caused by hypervirulent Acinetobacter baumannii
Chuchu LIN ; Min ZHANG ; Wenping ZHENG ; Wei HUANG
Chinese Journal of Clinical Infectious Diseases 2022;15(1):21-27
Objective:To trace and identify the source and the transmission vector of a nosocomial infection event caused by hypervirulent Acinetobacter baumannii. Methods:Nine strains of carbapenem-resistant Acinetobacter baumannii (CRAB) isolated from respiratory general ward and respiratory intensive care unit in Shenzhen People’s Hospital from July to August 2018 were collected. Environmental specimens such as ward environmental objects and medical staff’s hands were also collected. Vitek 2 was used for identification of bacterial species and drug susceptibility testing; Illuinam HiSeq 2500 platform, Staramr, Ridom SeqSphere+ were used for whole genome sequencing, drug resistance gene detection and multi-locus sequence typing (MLST); and IQ-Tree software, BEAST2 software package and SCOTTI software were used to construct phylogenetic tree and propagation path map. At the same time, the virulence of pathogenic bacteria was detected by mouse pneumonia model. Results:The 9 isolated CRAB strains were all ST2 type, all carried the carbapenem-resistant gene blaOXA-23, and the capsular types were KL49, KL3, KL77 and KL2, respectively. One of the CRAB strains isolated from removable ventilator screen was ST2 type and KL49 type capsule. Phylogenetic tree analysis showed that the environmental isolates were located in the same branch as 5 CRABs, and the transmission path map showed that these 5 CRABs were also located in the same transmission chain. Five strains of CRAB were involved in four lethal cases, and in vivo experiments in mice confirmed that the strains causing lethal infection had a hypervirulent phenotype. Conclusions:Highly virulent Acinetobacter baumannii can colonize the surface of mobile medical equipment, and sharing these equipment may lead to nosocomial spread of pathogens. The use of new propagation path analysis tools has certain reference significance for the investigation of propagation events.
7.Establishment of multi-point trigger and multi-channel surveillance mechanism for intelligent early warning of infectious diseases in China
Weizhong YANG ; Yajia LAN ; Wei LYU ; Zhiwei LENG ; Luzhao FENG ; Shengjie LAI ; Chuchu YE ; Qing WANG
Chinese Journal of Epidemiology 2020;41(11):1753-1757
This paper reviews the limitations of current infectious disease surveillance and early warning system in China, analyzes the concepts and countermeasures of the establishment of an intelligent early warning platform of infectious diseases based on multi-point trigger mechanism and multi-channel surveillance mechanism and proposes the realization routes for the purpose of facilitating capacity building and improvement of surveillance and early warning of infectious diseases in China.
8.Study on behavioral risk factors and lagging effect analysis with liver cancer mortality in rural critical areas of 4 provinces of China from 2009 to 2019
Xiaoying YANG ; Ning WANG ; Chuchu WEI ; Fengdie HE ; Jinlei QI ; Baohua WANG
Chinese Journal of Epidemiology 2023;44(10):1583-1590
Objective:To analyze the trend of liver cancer mortality in rural key areas of Jiangsu Province, Anhui Province, Shandong Province, and Henan Province (4 provinces) from 2009 to 2019 and to explore the influence of behavioral risk factors on liver cancer mortality and its lagging effect, and provide a reference for the prevention and treatment of liver cancer in China.Methods:Based on the 2009-2019 National Cause of Death Surveillance Database of the Chinese Center for Disease Control and Prevention, and the survey data of tumor and risk factor behavior of residents in key areas of 4 provinces, Joinpoint 4.2 software was used to calculate the average annual percentage change (AAPC) for assessing the temporal trend of standardized mortality of liver cancer; Chi-square test and trend Chi-square test were used to analyze the regional distribution difference and temporal change trend of behavioral habit factors. Stata 16 was used to establish a panel model to analyze the correlation and lagging effect of behavioral risk factors with liver cancer.Results:The standardized mortality rate of liver cancer in Jinhu County, Sheyang County, Lingbi County, Shou County, Mengcheng County, Wenshang County, Juye County, Luoshan County, Shenqiu County, and Xiping County showed a downward trend (AAPC<0, P<0.05) from 2009 to 2019. The consumption frequency of pickles/salted fish, red meat, and aquatic products showed a downward trend. The consumption frequency of healthy foods such as fresh vegetables, fresh fruits, and dairy products in all counties and districts showed an upward trend, and the consumption frequency of fried foods, kimchi, smoked foods, moldy foods, coffee, and soy products remained at a low level ( P<0.05); but the consumption frequency of soy products and dairy products was still <20.00%. Fried food, pickles/salted fish, current smoking rate, alcohol consumption rate, and unvaccinated hepatitis B vaccine rate were positively correlated with liver cancer death, and there was a lag effect, and the lag period was 4, 1, 6, 5, 4 years respectively. Conclusions:From 2009 to 2019, the mortality rate of liver cancer in rural key areas of 4 provinces shows a downward trend. There is a correlation and lagging effect between behavioral risk factors such as fried food, smoking, and alcohol consumption and liver cancer death.
9.Succinate/GPR91 promotes mitochondrial damage in vascular endothelial cells through DHODH/CoQ10
Wenhua QIN ; Chuchu YUAN ; Yuhui SUN ; Bo YU ; Dangheng WEI
Chinese Journal of Arteriosclerosis 2024;32(6):466-472
Aim To explore the effect of succinate/G protein coupled receptor 91(GPR91)on mitochondria in vascular endothelial cells and its regulatory mechanisms.Methods Transmission electron microscopy,Western blot and fluorescence microscopy were used to observe the effects of succinate analogues diethyl succinate(DS),GPR91 agonist and inhibitor on the mitochondrial morphology,cristae,cristate homeostasis related proteins reactive oxygen species(ROS)content,Ca2+concentration,mitochondrial membrane potential,the expression of dihydroorotate dehydrogenase(DHODH)and oxidized coenzyme Q10(CoQlO).Fluorescence probes were used to observe the effect of DHODH inhib-itor and CoQ10 on ROS level and Ca2+concentration of endothelial cells.Results After DS treatment,the mitochon-dria showed pyknosis and mitochondrial volume significantly decreased,electron density of the mitochondrial membrane in-creased,and the number of cristae decreased in endothelial cells;the expression of cristae homeostasis related proteins MIC60 decreased by 23%,while cellular ROS level and Ca2+concentration increased;mitochondrial membrane potential decreased(P<0.05 or P<0.01).After GPR91 agonist treatment,the expression of cristae homeostasis related proteins MIC60 decreased by 31%,meanwhile,cellular ROS level increased by 27%and Ca2+concentration increased by 36%;mitochondrial membrane potential decreased(P<0.05 or P<0.01).After GPR91 inhibitor treatment,the expression of cristae homeostasis related proteins MIC60 increased by 22%and ATP5I increased by 40%;the levels of ROS decreased by 41%and Ca2+concentration decreased by 67%;and the mitochondrial membrane potential was restored to normal(P<0.05 or P<0.01).After DS treatment,the expression of DHODH decreased by 43%and the level of oxidized CoQ10 in-creased by 120%(P<0.05 or P<0.01).After GPR91 agonist treatment,the expression of DHODH decreased by 22%and the level of oxidized CoQ10 increased by 36%(P<0.05 or P<0.01).After GPR91 inhibitor treatment,the expres-sion of DHODH increased by 40%and the level of oxidized CoQ10 decreased by 39%(P<0.01).After DHODH inhibi-tor treatment,the ROS level increased by 20%and Ca2+concentration increased by 28%,and mitochondrial membrane po-tential reduced at same time(P<0.05 or P<0.01).Exogenous oxidized CoQ10 inhibited ROS production by 30%and decreased Ca2+concentration by 20%(P<0.05 or P<0.01).Conclusion Succinate/GPR91 promotes mitochondrial damage in endothelial cells,and its mechanism may relate to down-regulating the expression of DHODH and inhibiting the reduction of CoQ10 by affecting the mitochondrial cristae homeostasis.