1.Correlation between blood pressure trajectory and hearing threshold among workers exposed to occupational noise in a city's rail transit enterprise
Hongting ZHAN ; Qia WANG ; Xinmei CHEN ; Zhiping LIANG ; Cong LI ; Danyan CAO ; Aichu YANG ; Minghui XIAO
Journal of Environmental and Occupational Medicine 2025;42(6):724-731
Background Hypertension is one of the chronic diseases with the highest prevalence in China, and a history of hypertension may potentially exacerbate hearing loss. Investigating the association between long-term blood pressure trends and hearing thresholds could contribute to hearing protection efforts for occupationally noise-exposed populations. Objective By investigating hearing thresholds and blood pressure levels among occupationally noise-exposed workers in an urban rail transit enterprise, and conducting a comprehensive analysis of the association between long-term blood pressure changes and hearing thresholds, to provide data references for health management strategies targeting occupationally noise-exposed workers. Methods Workers exposed to occupational noise at a rail transit enterprise were enrolled as study subjects and underwent pure-tone audiometry. Group-based trajectory modeling was employed to identify blood pressure trajectories. Categorical data were compared using chi-square tests, while normally distributed continuous variables were analyzed via t-tests and analysis of variance (ANOVA). Generalized linear mixed models (GLMMs) were subsequently applied toexamine associations between these trajectory groups and high-frequency hearing thresholds. Results Among 2 002 occupationally noise-exposed workers, the median (P25, P75) age was 32 (28, 35) years, with a median (P25, P75) working tenure of 7 (3, 10) years. In 2019, the positive hypertension rate was 9.04%, with a mean systolic blood pressure (SBP) of (122.97±11.60) mmHg and a mean diastolic blood pressure (DBP) of (76.37±9.02) mmHg. The hearing loss prevalence was 10.1%, showing bilateral high-frequency average hearing thresholds of (17.18±8.71) dB and speech-frequency average thresholds of (13.79±3.46) dB. Three distinct trajectory groups were identified for both SBP and DBP. Compared with other trajectory groups, the high-stable DBP group exhibited significantly higher hearing loss prevalence (χ2=6.34, P=0.042) and elevated high-frequency hearing thresholds (all Ps<0.05). Specifically, within the 30-39 age subgroup, the moderate-stable DBP group demonstrated 1.96 dB lower high-frequency thresholds than the high-stable group [β(95%CI): −1.96 (−3.61, −0.32), P=0.020]. Conclusion Among occupationally noise-exposed workers in a municipal rail transit enterprise, DBP trajectories demonstrated a positive association with high-frequency hearing thresholds. Notably, in young and middle-aged occupationally noise-exposed populations, DBP may exert a more critical influence than SBP on the progression of hearing loss.
2.Construction and application of a whole-process intelligent management system for preventing PICC-related bloodstream infections
Enci LI ; Mudan WANG ; Lianguo CHEN ; Huijie ZHOU ; Danyan HU ; Yiren HU
Chinese Journal of Modern Nursing 2024;30(10):1364-1368
Objective:To construct the whole-process intelligent management system for preventing PICC-related bloodstream infections and explore its effect in preventing PICC-related bloodstream infections.Methods:From January 2021 to December 2022, patients with PICC admitted to Wenzhou People' s Hospital were selected as the research subject, the patients from January to December 2021 were divided into the control groups, the patients from January to December 2022 were divided into the observation group. The whole-process intelligent management system for preventing PICC-related bloodstream infections was constructed and applied with artificial intelligence, this study compared the incidence of PICC-related bloodstream infections and the implementation rate of PICC whole-process bundled management projects before and after system application.Results:The incidence of PICC-related bloodstream infections in the control group was 0.55‰ (14/25 674), while the incidence in the observation group was 0.20‰ (5/25 226), with a statistically significant difference (χ 2=4.110, P<0.05). The implementation rates of PICC whole-process bundled management projects in the control group and observation group were 74.04% (2 319/3 132) and 92.11% (2 885/3 132), respectively, with a statistically significant difference (χ 2=363.782, P<0.01) . Conclusions:The whole-process intelligent management system for preventing PICC-related bloodstream infections constructed optimizes the prevention and treatment process of PICC-related bloodstream infections, effectively ensures the implementation of nursing interventions and monitoring measures, and reduces the incidence of PICC-related bloodstream infections.
3.Hemodynamics and arrhythmia disorder caused by lithium poisoning:A case report
Yujue WANG ; Lijun OUYANG ; Xiaoqian MA ; Danyan XU ; Xiaogang CHEN ; Ying HE
Journal of Central South University(Medical Sciences) 2024;49(1):153-158
Bipolar affective disorder refers to a category of mood disorders characterized clinically by the presence of both manic or hypomanic episodes and depressive episodes.Lithium stands out as the primary pharmacological intervention for managing bipolar affective disorder.However,its therapeutic dosage closely approaches toxic levels.Toxic symptoms appear when the blood lithium concentration surpasses 1.4 mmol/L,typically giving rise to gastrointestinal and central nervous system reactions.Cardiac toxicity is rare but serious in cases of lithium poisoning.The study reports a case of a patient with bipolar affective disorder who reached a blood lithium concentration of 6.08 mmol/L after the patient took lithium carbonate sustained-release tablets beyond the prescribed dosage daily and concurrently using other mood stabilizers.This resulted in symptoms such as arrhythmia,shock,impaired consciousness,and coarse tremors.Following symptomatic supportive treatment,including blood dialysis,the patient's physical symptoms gradually improved.It is necessary for clinicians to strengthen the prevention and recognition of lithium poisoning.
4.Effectiveness and security of anisodine hydrobromide tablets in treating nonarteritic anterior ischemic optic neuropathy: a Chinese multicenter nonrandomized controlled study
Mo YANG ; Honglu SONG ; Huanfen ZHOU ; Mengying LAI ; Quangang XU ; Mingming SUN ; Ke FAN ; Hongpei CUI ; Haiyan WANG ; Xin JIN ; Chuanbin SUN ; Qing XIAO ; Ying WANG ; Zide ZHAO ; Minglian ZHANG ; Yongye CHANG ; Mengping CHEN ; Zhanxing SHEN ; Hui YANG ; Xiaoyu XU ; Zhiqing LI ; Dongjun XING ; Yu DONG ; Jinrun YANG ; Qian REN ; Li LI ; Wenfang ZHANG ; Li SUN ; Zhengpei ZHANG ; Suyan LI ; Danyan LIU ; Nalei ZHOU ; Nali LUO ; Yadong LIU ; Shihui WEI
Chinese Journal of Experimental Ophthalmology 2023;41(7):646-653
Objective:To evaluate the efficacy and safety of oral anisodine hydrobromide tablets in the treatment of nonarteritic anterior ischemic optic neuropathy (NAION).Methods:A multicenter nonrandomized controlled trial was conducted.A total of 282 acute NAION patients (282 eyes) were recruited from 16 hospitals in China from July 2020 to May 2021.Patients were divided into two groups according to treatment methods, which were control group (124 cases, 124 eyes) receiving regular treatment including citicoline sodium plus Ginkgo biloba leaf liquid extract or Ginkgo biloba leaf extract tablets plus mecobalamin, and experimental group (158 cases, 158 eyes) receiving treatment in control group plus oral anisodine hydrobromide tablets 1 mg, twice daily for 2 to 3 months.Best corrected visual acuity (BCVA), visual field index (VFI), peripapillary retinal nerve fiber layer (pRNFL) and radial peripapillary capillary vessel density (RPC) were assessed at 1, 2, 3, and 6 months after enrollment using the standard decimal visual acuity chart, 750i Humphery visual field analyzer, Cirrus HD-OCT 4000/Cirrus HD-OCT 5000, RTVue-XR optical coherence tomography respectively.The primary outcomes were BCVA and VFI, and the secondary outcomes were pRNFL, RPC, and the side effects during the follow-up.The study adhered to the Declaration of Helsinki.All patients were fully informed about the treatment and purpose of this study and voluntarily signed the informed consent form.The study protocol was approved by Chinese PLA General Hospital (No.S2020-021-01). Results:In all, 242 patients (242 eyes) completed the follow-up of BCVA, and 98 patients (98 eyes) completed the VFI follow-up.In terms of visual function, BCVA and VFI improved significantly over time in the two groups, and BCVA and VFI were better in experimental group than in control group at various follow-up time points (all at P<0.05). In terms of structure, pRNFL gradually decreased in both groups with the extension of treatment, and pRNFL was significanthy thinner in experimental group than in control group at various follow-up time points (all at P<0.05). There was no significant difference in RPC between the two groups at the last follow-up ( P>0.05). There were two cases with side effects and one case was discontinued due to side effects 25 days after enrollment. Conclusions:Oral anisodine hydrobromide can improve visual acuity and visual field in NAION and accelerate the regression of optic disc edema, with good safety.
5.A prospective study of genetic screening of 2 060 neonates by high-throughput sequencing.
Danyan ZHUANG ; Fei WANG ; Shuxia DING ; Zhoushu ZHENG ; Qi YU ; Lanqiu LYU ; Shuni SUN ; Rulai YANG ; Wenwen QUE ; Haibo LI
Chinese Journal of Medical Genetics 2023;40(6):641-647
OBJECTIVE:
To assess the value of genetic screening by high-throughput sequencing (HTS) for the early diagnosis of neonatal diseases.
METHODS:
A total of 2 060 neonates born at Ningbo Women and Children's Hospital from March to September 2021 were selected as the study subjects. All neonates had undergone conventional tandem mass spectrometry metabolite analysis and fluorescent immunoassay analysis. HTS was carried out to detect the definite pathogenic variant sites with high-frequency of 135 disease-related genes. Candidate variants were verified by Sanger sequencing or multiplex ligation-dependent probe amplification (MLPA).
RESULTS:
Among the 2 060 newborns, 31 were diagnosed with genetic diseases, 557 were found to be carriers, and 1 472 were negative. Among the 31 neonates, 5 had G6PD, 19 had hereditary non-syndromic deafness due to variants of GJB2, GJB3 and MT-RNR1 genes, 2 had PAH gene variants, 1 had GAA gene variants, 1 had SMN1 gene variants, 2 had MTTL1 gene variants, and 1 had GH1 gene variants. Clinically, 1 child had Spinal muscular atrophy (SMA), 1 had Glycogen storage disease II, 2 had congenital deafness, and 5 had G6PD deficiency. One mother was diagnosed with SMA. No patient was detected by conventional tandem mass spectrometry. Conventional fluorescence immunoassay had revealed 5 cases of G6PD deficiency (all positive by genetic screening) and 2 cases of hypothyroidism (identified as carriers). The most common variants identified in this region have involved DUOX2 (3.93%), ATP7B (2.48%), SLC26A4 (2.38%), GJB2 (2.33%), PAH (2.09%) and SLC22A5 genes (2.09%).
CONCLUSION
Neonatal genetic screening has a wide range of detection and high detection rate, which can significantly improve the efficacy of newborn screening when combined with conventional screening and facilitate secondary prevention for the affected children, diagnosis of family members and genetic counseling for the carriers.
Child
;
Infant, Newborn
;
Humans
;
Female
;
Prospective Studies
;
Connexins/genetics*
;
Connexin 26/genetics*
;
Glucosephosphate Dehydrogenase Deficiency
;
Mutation
;
Sulfate Transporters/genetics*
;
DNA Mutational Analysis
;
Genetic Testing/methods*
;
Deafness/genetics*
;
Neonatal Screening/methods*
;
Hearing Loss, Sensorineural/genetics*
;
High-Throughput Nucleotide Sequencing
;
Solute Carrier Family 22 Member 5/genetics*
6.Application value of transrectal multimodal ultrasound in evaluating the efficacy of endocrine therapy for prostate cancer
Qian LIU ; Hua HONG ; Danyan LIANG ; Guozhu WU ; Ran SUN ; Fang WANG ; Yue SUN ; Yuan FENG
Chinese Journal of Ultrasonography 2022;31(9):802-808
Objective:To explore the value of transrectal multimodal ultrasound parameters in monitoring and evaluating the efficacy of endocrine therapy for prostate cancer.Methods:Thirty patients with prostate cancer confirmed by pathology and treated with endocrine therapy in Inner Mongolia Autonomous Region People′s Hospital from November 2019 to May 2021 were selected. The levels of serum prostate specific antigen (PSA), prostate volume, color Doppler parameters, elasticity index and contrast-enhanced ultrasound parameters were measured and recorded before treatment, 1 month and 3 months after treatment. The parameters before and after treatment were statistically analyzed. The correlation between the changes of each index and PSA was analyzed by Spearman correlation analysis.Results:Total prostate specific antigen, free prostate specific antigen, and prostate volume were significantly different before treatment, and 1 month and 3 months after treatment( P<0.05), and the values showed a downward trend with increase of treatment time. There was no significant difference in resistance index before and 1 month after treatment( P>0.05), but decreased significantly 3 months after treatment( P<0.05). The values of elasticity index, peak intensity, area under curve and gradient at 1 month and 3 months after treatment were lower than those before treatment, while the arrival time and rising time at 1 month and 3 months after treatment were significantly higher than those before treatment( P<0.05). Spearman correlation analysis showed that there was no correlation between the changes of quantitative parameters and PSA value before and after treatment( P>0.05). Conclusions:Prostate volume, color Doppler parameters, elasticity index, and contrast-enhanced ultrasound parameters change in the early stage of endocrine therapy for prostate cancer, which can be used as a useful supplement to PSA for prostate cancer, and can be used to evaluate the efficacy of clinical prostate cancer endocrine therapy.
7.The role of coupler perfusion in transrectal ultrasound in the diagnosis of T-stage rectal cancer
Guozhu WU ; Yanwei GAO ; Hua HONG ; Yumin WANG ; Xianyan MENG ; Ran SUN ; Qian LIU ; Haijun LI ; Danyan LIANG
Chinese Journal of Ultrasonography 2021;30(2):151-156
Objective:To investigate the role of coupler perfusion in transrectal ultrasound in the diagnosis of preoperative T staging of rectal cancer.Methods:A retrospective analysis of the preoperative clinical data of 132 patients with rectal cancer in the People′s Hospital of Inner Mongolia Autonomous Region from June 2015 to November 2020. According to whether or not the patients agreed to coupler perfusion before ultrasound examination, they were divided into 2 groups, namely the perfusion group 69 cases and the non-perfusion group of 63 cases, with postoperative pathology as the gold standard, and compared with magnetic resonance imaging(MRI) to evaluate the accuracy of the 2 groups and MRI in the T staging of rectal cancer.Results:The total coincidence rates of the coupling agent perfusion group, non-perfusion group and MRI group for the diagnosis of rectal cancer T staging were 89.9%, 76.2% and 87.9%, respectively, and the difference among the three methods was statistically significant (χ 2=6.096, P=0.047). The diagnostic sensitivity of the coupling agent perfusion group for T1 stage was 96.0%, which was higher than 61.5% of the non-perfusion group and 92.3% of the MRI ( P=0.010). The specificity of the perfusion group for the diagnosis of T2 stage was 95.7%, higher than the non-perfusion group and MRI ( P=0.037), the positive predictive value of the perfusion group for T2 stage was 90.9%, which was higher than the non-perfusion group and MRI ( P=0.035). The diagnostic accuracy of the perfusion group for T2 stage was 94.2%, higher than the non-perfusion group and MRI (χ 2=7.070, P=0.029). There were no statistically significant differences in diagnostic sensitivity, specificity, positive predictive value, negative predictive value and accuracy among the perfusion group and the non-perfusion group and the MRI for T3 and T4 (all P>0.05). Conclusions:Coupled-agent perfusion makes it convenient and fast for intracavity ultrasound to diagnose T staging of rectal cancer, and the diagnostic efficiency is comparable to MRI. In particular, it can be used as a highly reliable imaging method for T1 and T2 rectal cancer.
8.Analysis of diagnostic efficacy of targeted biopsy versus targeted biopsy combined with systematic biopsy for patients with PI-RADS score of 4-5
Yu LIU ; Jie GAO ; Wei WANG ; Qing ZHANG ; Xiaozhi ZHAO ; Haifeng HUANG ; Danyan LI ; Yao FU ; Hongqian GUO
Chinese Journal of Urology 2021;42(3):192-196
Objective:To analyze the diagnostic efficacy of targeted biopsy (TB) versus targeted biopsy combined with systematic biopsy (TB+ SB) for patients with multi-parametric magnetic resonance imaging (mpMRI) prostate imaging-reporting and data system (PI-RADS) score of 4-5.Methods:The clinical data of 378 patients with mpMRI PI-RADS score of 4-5 in Nanjing Drum Tower Hospital from January 2018 to February 2020 who received prostate TB+ SB were retrospectively analyzed. Median age was 69 (64, 75) years old, median prostate specific antigen was 9.5 (6.7, 16.3) ng/ ml, and median prostate volume was 34.1 (23.5, 48.4) ml. There were 240 cases with PI-RADS score of 4 and 138 cases with PI-RADS score of 5. Evaluating Gleason score of positive biopsy pathology and using χ 2 test or Fisher exact test to analyze the detection of prostate cancer (PCa) and clinically significant prostate cancer(CsPCa) by TB versus TB+ SB. Results:Of the all 378 cases, 88 cases (23.3%) were negative and 290 cases (76.7%) were positive. The average number of needle for TB was 2.4 per person, while SB was 12 per person. TB and SB had no statistically significant difference in the detection rate of PCa (73.3% vs. 68.3%, P=0.129) and CsPCa (55.8% vs. 49.7%, P=0.094) and in the accuracy (79.1% vs. 77.8%, P=0.658), but had a statistically significant difference in the positive rate (64.2% vs. 23.1%, P < 0.001). The pathological coincidence rate of TB and TB+ SB was 92.3%. There was no statistical difference in the detection rate of PCa (73.3% vs. 76.7%, P=0.275) and CsPCa (55.8% vs. 62.2%, P=0.076) between TB and TB+ SB. The missed diagnosis rate of TB for PCa was 4.5%, for CsPCa was 10.2%. For patients with PI-RADS score of 4, TB had no significant difference in the detection rate of PCa (65.4% vs. 69.2%, P=0.381) and CsPCa (46.7% vs. 52.9%, P=0.171) from TB+ SB. The accuracy of TB was 82.1%. The missed diagnosis rate of TB for PCa was 5.4%, for CsPCa was 11.8%. For patients with PI-RADS score of 5, TB had no significant difference in the detection rate of PCa (87.0% vs. 89.9%, P=0.452) and CsPCa (71.7% vs. 78.3%, P=0.211) from TB+ SB. The accuracy of TB was 73.9%. The missed diagnosis rate of TB for PCa was 3.2%, for CsPCa was 8.3%. Conclusions:For high-risk prostate cancer patients with PI-RADS score of 4-5, TB can obtain a detection effect similar to that of TB+ SB with fewer needles, but there is still the possibility of inaccurate diagnosis and missed diagnosis.
9.Experimental study on the effect of three-dimensional porous structures on the vascularization rate of artificial dermis
Rongwei TAN ; Xi LIU ; Yingying CHEN ; Mengqiang XU ; Yuanjun GUO ; Danyan WANG ; Jiamei LIANG ; Jiao LIU ; Shasha YUAN ; Wei FAN ; Xiangkun WANG ; Zhending SHE
Chinese Journal of Burns 2021;37(10):959-969
Objective:To explore the effects of orienting three-dimensional porous network (type A) and honeycomb briquette-shaped vertically penetrating three-dimensional porous network (type B) on the vascularization rate of artificial dermis.Methods:The experimental research method was used. The artificial dermis was composed of a double layer of silicone layer and scaffold layer. Based on the difference of scaffold layer, they were divided into type A and type B artificial dermis (type A dermis and type B dermis, for short) containing type A and type B structure, respectively. The type A and type B structures were prepared by gradient freeze-drying technique and physical pore-making technique, respectively. The micro-morphology of two kinds of dermis scaffold was observed by scanning electron microscopy. The porosity of two kinds of dermis scaffold was measured by the Pyrex method. According to the method of national medical industry standard, the hydroxyproline content in degradation liquids and their residues in two kind of dermis were determined after degradation at 4, 8, 13, and 24 h, reflecting the degradation rates of two kinds of dermis. According to the random number table, L929 cells were divided into type A dermis group, type B dermis group, negative control group, and positive control group. The positive control group was added with minimum essential medium (MEM) containing 5% dimethyl sulfoxide, The negative control group was added with high-density polyethylene extract, and the other two groups were added with the corresponding extract. At 24 hours after culture, the growth rate of L929 cells was detected by methyl thiazolyl tetrazolium, and the cytotoxicity was graded. L929 cells and human umbilical vein endothelial cells (HUVECs) were inoculated into pore plates with two kinds of dermis preinstalled. On 1, 4, 7, and 14 d after inoculating, the adhesion and growth of L929 cells on the surfaces of the two kinds of scaffolds were detected by immunofluorescence method. On 7 d after inoculating, the migration of the above two kinds of cells into the two kinds of dermal scaffolds was detected by immunofluorescence and hematoxylin-eosin (HE) staining. Three full-thickness skin defect wounds of 5.0 cm×5.0 cm were created on both sides of the back of three 6-month-old healthy male Ba-Ma mini pigs. According to the random number table, six columns of wounds were divided into type A dermis two-step method group, type B dermis two-step method group, and type B dermis one-step method group. The wounds in type A dermis two-step method group and type B dermis two-step method group were transplanted with type A or type B dermis respectively before, and with autologous split-thickness skin grafting later. The wounds in type B dermis one-step method group were transplanted in a synchronous procedure including type B dermis (without silicone layer) and autologous skin grafting simultaneously. The bleeding, exudation, and infection of the wounds on the back in type A dermis two-step method group and type B dermis two-step method group on the 7th day after the second transplantation and in type B dermis one-step method group on the 14th day after the first transplantation were generally observed. The area of autologous skin graft was measured by the transparent film grid method, and the survival rate of autologous skin was calculated. On 4, 7, and 14 d after the first transplantation, the inflammatory cells, fibroblasts (Fbs), and capillary infiltration into the scaffolds of the three groups were detected by HE staining. On 7, 14 d after the first transplantation, the vascularization of the scaffolds was further observed by immunohistochemistry. On 28, 90 d after the first operation, the degradation of the scaffolds of type A dermis and type B dermis was observed by HE staining. Data were statistically analyzed with one-way analysis of variance, independent sample t test, and Bonferroni correction. Results:A large number of round and oval micropores were evenly distributed on the surface of type A scaffold, and the cylindrical hole walls could be observed arranging in a parallel direction in the longitudinal section. The honeycomb briquette-shaped penetrating macropores on the surface of type B scaffold were arranged in an orderly matrix. The pore walls of the honeycomb briquette-shaped penetrating macropores were connected by micropores to form a network structure. The porosity of type A dermis was (93.21±0.72)%, which was similar to (95.88±1.00)% of type B dermis ( t=4.653, P>0.05). The degradation rates of type A dermis at 4, 8, 13, and 24 h were similar to those of type B dermis at the corresponding time point ( t=0.232, 0.856, 0.258, 7.716, P>0.05). At 24 h after culture, the proliferation rates of L929 cells in the type A dermis group, type B dermis group, and negative control group were significantly higher than those of the positive control group ( t=2 393.46, 2 538.27, 1 077.77, P<0.01). The cytotoxicity rating of cells in positive control group was grade 4, while that of the other three groups was grade zero. On 1, 4, 7, and 14 d after inoculation, both L929 cells and HUVECs proliferated in a time-dependent manner in two kinds of dermal scaffolds. The adhesion growth and proliferation rate of the two kinds of cells on the surface of type B dermis was higher than that of type A dermis. On 7 d after inoculation, both L929 cells and HUVECs covered the surface of type B dermis and migrated into one side of the silicone layer. However, the above two kinds of cells migrated slowly into type A dermis, and only a few cells were found on one side of the silicone layer. There was no bleeding, exudation, or infection in the wounds repaired by type A and type B dermis. The survival rate of autologous skin grafting of 6 wounds in each group was 100%. On 4, 7, and 14 d after the first operation, inflammatory cells, Fbs, and capillaries gradually infiltrated into the scaffold layer, and the cell infiltration rate from high to low was type B dermis one-step method group, type B dermis two-step method group, and type A dermis two-step method group. The scaffold in wound in the type B dermis one-step method group gradually collapsed on 28 d after the first operation, and completely degraded in 3 months after the first operation. The scaffold degradation rate of type A dermis two-step method group was similar to that mentioned above. Conclusions:The honeycomb briquette-shaped vertically penetrating three-dimensional porous network structure of type B scaffold can accelerate its vascularization process, which is beneficial to autogenous split-thickness skin in one-step procedure to repair full-thickness skin defects wound in Ba-Ma mini pigs. Compared with the "two-step method" of staged transplantation of type A scaffold and autologous split-thickness skin, and one-step transplantation has equal efficacy and can provide a better choice for wound treatment.
10.Rictor regulates mitochondrial calcium signaling in mouse embryo stem cell-derived cardiomyocytes.
Ying SHAO ; Jiadan WANG ; Danyan ZHU
Journal of Zhejiang University. Medical sciences 2019;48(1):65-74
OBJECTIVE:
To explore the expression, localization and regulatory effect on mitochondrial calcium signaling of Rictor in embryonic stem cell-derived cardiomyocytes (ESC-CMs).
METHODS:
Classical embryonic stem cell cardiomyogenesis model was used for differentiation of mouse embryonic stem cells into cardiomyocytes. The location of Rictor in ESC-CMs was investigated by immunofluorescence and Western blot. The expression of Rictor in mouse embryonic stem cells was interfered with lentiviral technology, then the superposition of mitochondria and endoplasmic reticulum (ER) in ESC-CMs was detected with immunofluorescence method; the cellular ultrastructure of ESC-CMs was observed by transmission electron microscope; the mitochondrial calcium transients of ESC-CMs was detected by living cell workstation;immunoprecipitation was used to detect the interaction between 1,5,5-trisphosphate receptor (IP3 receptor, IP3R), glucose-regulated protein 75 (Grp75) and voltage-dependent anion channel 1 (VDAC1) in mitochondrial outer membrane; the expression of mitochondrial fusion protein (mitonusin-2, Mfn2) was detected by Western blot.
RESULTS:
Rictor was mainly localized in the endoplasmic reticulum and mitochondrial-endoplasmic reticulum membrane (MAM) in ESC-CMs. Immunofluorescence results showed that Rictor was highly overlapped with ER and mitochondria in ESC-CMs. After mitochondrial and ER were labeled with Mito-Tracker Red and ER-Tracker Green, it was demonstrated that the mitochondria of the myocardial cells in the Rictor group were scattered, and the superimposition rate of mitochondria and ER was lower than that of the negative control group (<0.01). The MAM structures were decreased in ESC-CMs after knockdown of Rictor. The results of the living cell workstation showed that the amplitude of mitochondrial calcium transients by ATP stimulation in ESC-CMs was decreased after knockdown of Rictor (<0.01). The results of co-immunoprecipitation showed that the interaction between IP3R, Grp75 and VDAC1 in the MAM structure of the cardiomyocytes in the Rictor group was significantly attenuated (<0.01); the results of Western blot showed that the expression of Mfn2 protein was significantly decreased (<0.01).
CONCLUSIONS
Using lentiviral technology to interfere Rictor expression in mouse embryonic stem cells, the release of calcium from the endoplasmic reticulum to mitochondria in ESC-CMs decreases, which may be affected by reducing the interaction of IP3R, Grp75, VDAC1 and decreasing the expression of Mfn2, leading to the damage of MAM structure.
Animals
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Calcium Signaling
;
genetics
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Gene Expression Regulation
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genetics
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Gene Knockdown Techniques
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Mice
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Mitochondria
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physiology
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Mouse Embryonic Stem Cells
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Myocytes, Cardiac
;
physiology
;
Protein Transport
;
Rapamycin-Insensitive Companion of mTOR Protein
;
genetics
;
metabolism

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