1.Effects of di(2-ethylhexyl) phthalate on glucose homeostasis in rats due to impaired autophagy flux of islet β cells mediated by oxidative stress
Hongyang ZHOU ; Yuting HU ; Xue CHEN ; Yunqiang ZHOU ; Liping LI ; Ling LI ; Herong LIU
Journal of Environmental and Occupational Medicine 2025;42(6):674-683
Background Di(2-ethylhexyl) phthalate (DEHP) is the most prevalent environmental endocrine disruptor among phthalate acid esters (PAEs) worldwide. Previous studies have indicated that exposure to DEHP may disrupt glucose metabolism. Objective To investigate the impact of DEHP on glucose homeostasis in rats, focusing on oxidative stress-induced impairment of autophagy in islet β cells. Methods Forty male SD rats were randomly assigned to four groups, receiving DEHP doses of 0, 187, 375, and 750 mg·kg−1 for 12 weeks. Oral glucose tolerance (OGTT) and insulin tolerance tests (ITT) were conducted 24 h after the final exposure. Pancreatic microstructural alterations were assessed using hematoxylin and eosin (HE) staining and transmission electron microscopy (TEM). Commercial ELISA kits were employed to quantify the levels of insulin, adenosine triphosphate (ATP), and adenosine monophosphate (AMP) in rat serum, as well as the protein expression level of activated caspase-3 in pancreatic tissue. Additionally, commercial microplate kits were utilized to measure the concentration of reduced glutathione (GSH) in serum, the activity of superoxide dismutase (SOD) using water-soluble tetrazolium salt-1, the content of malondialdehyde (MDA) by thiobarbituric acid method, and the level of reactive oxygen species (ROS) in pancreatic tissue by chemical fluorescence method. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure sequestosome1 (SQSTM1/p62), Beclin1, microtubule-associated protein 1 light chain 3 (LC3), and cysteinyl aspartate specific proteinase-8 (Caspase-8) mRNA levels. Western blot analysis was applied to detect the protein relative expression levels of p62, Beclin-1, LC3-I, LC3 II, AMPK, p-AMPK, mTOR, p-mTOR, ULK1, and Caspase-8. Results Compared to the 0 mg·kg−1 DEHP group, the 750 mg·kg−1 DEHP group exhibited a significant increase in fasting blood glucose levels at 2, 4, 6, and 12 weeks (P<0.05). The OGTT showed that, following high-glucose gavage, the 187 mg·kg−1 DEHP group had elevated blood glucose at 30 min (P<0.05), the 375 mg·kg−1 DEHP group showed increased glucose levels at 15, 30, and 180 min (P<0.05), and the 750 mg·kg−1 DEHP group exhibited elevated levels at 15, 30, 60, and 180 min (P<0.05). The 375 and 750 mg·kg−1 DEHP groups demonstrated significantly increased OGTT area under the curve (AUC) values (P<0.05). In contrast, ITT results indicated no significant differences in blood glucose levels or AUC among the DEHP exposure groups at all time points (P>0.05). Compared to the 0 mg·kg−1 DEHP group, the 750 mg·kg−1 DEHP group exhibited significantly higher HOMA-IR levels and markedly lower HOMA-ISI values (P<0.05). HE and TEM showed that in each DEHP exposure group, the number of islet cells decreased, the islet area reduced, and chromatin condensation occurred. The endocrine granules in the cytoplasm of islet β cells decreased, and there were varying degrees of widening of the nuclear membrane gap, flattening and expansion of the Golgi complex, and expansion of the endoplasmic reticulum. Ribosome separation was observed, and autophagosomes were visible. In the 375 and 750 mg·kg−1 DEHP groups, the mitochondria were deformed to varying degrees, and some cristae structures disappeared, presenting vacuolization. Moreover, the chromatin condensation in the nuclei was more severe in the 750 mg·kg−1 DEHP group. The serum SOD activity was significantly elevated in the 750 mg·kg−1 DEHP group (P<0.05). Both the 375 mg·kg−1 and 750 mg·kg−1 DEHP groups exhibited a significant increase in the relative ROS content in pancreatic tissue (P<0.05). In DEHP-treated groups, the MDA content increased (P<0.05), while the GSH content decreased (P<0.05). Additionally, in the 750 mg·kg−1 DEHP group, the AMP/ATP ratio in serum was significantly raised (P<0.05), and the expression of cleaved Caspase-3 protein in pancreatic tissue was also significantly increased (P<0.05). The relative mRNA levels of p62, Beclin-1, LC3, and Caspase-8 in the pancreatic tissue of rats exposed to DEHP were significantly elevated (P<0.05). The relative expression levels of p-AMPK/AMPK, p-ULK1/ULK1, and Beclin-1 proteins in the DEHP-treated groups were significantly increased (P<0.05). In the 375 mg·kg−1 and 750 mg·kg−1 DEHP treatment groups, the relative expression levels of p62, LC3 II/LC1, and Caspase-8 proteins were significantly increased (P<0.05), while the relative expression level of p-mTOR/mTOR was significantly decreased (P<0.05). Conclusion DEHP can disrupt glucose homeostasis by inducing oxidative stress, which subsequently activates autophagy via the ROS/AMPK/ULK1 pathway, impairing autophagic flux and promoting apoptosis of islet β cells, ultimately decreasing their function and number.
2.Expression of IL-18,IL-18BP,and IL-18R in peripheral blood B lymphocytes and monocytes of patients with atopic dermatitis
Huitong LI ; Luning REN ; Fei WANG ; Dongmei YANG ; Hongyang DU
Journal of China Medical University 2024;53(9):782-787
Objective To investigate the expression of IL-18,IL-18BP,and IL-18R in the peripheral blood B lymphocytes and mono-cytes of patients with atopic dermatitis(AD).Methods Peripheral venous blood was collected from 28 patients with AD and 21 healthy controls,and stimulated with Artemisia sieversiana wild allergen extract,house dust mite allergen extract,or Platanus pollen allergen extract.The expression of IL-18+,IL-18BP+,and IL-18R+in B lymphocytes and monocytes was measured using flow cytometry.Results Compared with the healthy control group,the proportions of IL-18+,IL-18BP+,and IL-18R+cells in the B lymphocyte group of patients with AD at rest increased 2.01-,10.35-,and 20.85-fold,respectively.The proportions of IL-18+and IL-18BP+cells in monocytes increased 5.51-and 41.88-fold,respectively,whereas the proportion of IL-18R+cells did not differ significantly between the groups.Conclusion IL-18,IL-18BP,and IL-18R in B lymphocytes and monocytes may play an important role in AD.IL-18,IL-18BP,and IL-18R may be potential targets for the treatment of AD.
3.Role of UBC9-mediated SUMO modification in homocysteine-induced pyroptosis of macrophages
Lingju MA ; Hongyang CHI ; Xinxue WU ; Fujun MA ; Yancheng TIAN ; Caiqi ZHAO ; Tianyu HE ; Hongjian PENG ; Yideng JIANG ; Li YANG ; Hui HUANG ; Shengchao MA
Chinese Journal of Comparative Medicine 2024;34(6):11-17
Objective To study the role of ubiquitin-conjugating enzyme 9(UBC9)in the pyroptosis of homocysteine-induced macrophages mediated by small ubiquitin-like modifier(SUMO)modification.Methods First,the effects of homocysteine at different concentrations(0 μmol/L,50 μ.mol/L,100 μmol/L,150 μmol/L and 200 μmol/L)on the viability and pyrodeath of mouse macrophages(RAW264.7)were detected by CCK-8 and Western blot.Western blot was used to detect the expression levels of UBC9,SUMO-1,and the inflammatory cytokine IL-1β in different groups of cells.qRT-PCR was used to detect the mRNA expression of UBC9 before and after RNA interference and the expression of UBC9,pyrogen-related protein,and SUMO-1 after RNA interference.Results After stimulation with 100 μmol/L homocysteine,the effect of macrophage activity was minimal,and NLRP3 and Caspase-1 were the proteins with the most obvious increase in expression(P<0.05).Compared with the Control group,the Hcy group's expression of IL-1β and SUMO-1 was increased(P<0.01).Compared with the Control group,the Hcy group's UBC9 protein and mRNA levels were increased(P<0.05).The expression of NLRP3,Caspase-1,IL-1β,UBC9,and SUMO-1 was decreased in the si-UBC9+Hcy group compared with the si-NC+Hcy group(P<0.01).Conclusions Homocysteine induces pyroptosis in macrophages,and its mechanism of action is related to the up-regulation of UBC9 to induce SUMO modification.
4.Application Effect and Prognosis Analysis of D-J Tube in Pediatric Pyeloplasty—Propensity Score Matching
Hongyang WANG ; Jinqiu SONG ; Long LI
Journal of Medical Research 2024;53(10):93-98
Objective To analyze the indications and prognosis of the D-J tube internal drainage method in pediatric pyeloplasty surgery.Methods We conducted a retrospective analysis of cases of unilateral pyeloplasty performed in the urology department of the Capital Institute of Pediatrics from January 2012 to January 2022.They were divided into the D-J tube group(396 cases)and the py-elostomy group(334 cases)according to different postoperative drainage methods.The propensity score matching method was used to e-venly match the baseline data of the two groups of cases,and the complications and prognosis of the two drainage methods were compared.Results D-J tube drainage could significantly reduce the average hospitalization time,reduce pain,alleviate skin scars,and improve the quality of medical care and care.D-J tube could increase the risk of postoperative urinary tract infection and re-obstruction in chil-dren,and clinical application should be cautious.A predictive model was established for urinary tract infections after D-J tube implanta-tion,including gender,side profile,height,renal parenchymal thickness,and blood urea nitrogen.Conclusion The D-J tube internal drainage method can significantly reduce the length of hospital stay and nursing burden after pediatric pyeloplasty surgery.Although it may increase the risk of urinary tract infection in the near and medium term,the risk of urinary tract infection can be predicted.The use of D-J tubes should be carefully selected in males,small infants,and severe hydronephrosis.
5.Role of specific lncSLC25a6 in homocysteine-induced cuproptosis in rat cardiomyocytes
Shujuan LI ; Hui HUANG ; Hongyang CHI ; Lexin WANG ; Tianyu HE ; Fu-Jun MA ; Yancheng TIAN ; Caiqi ZHAO ; Hongjian PENG ; Yideng JIANG ; Li YANG ; Shengchao MA
Chinese Journal of Pathophysiology 2024;40(8):1399-1407
AIM:To investigate the role of specific long noncoding RNA SLC25a6(lncSLC25a6)in homocys-teine(Hcy)-induced cuproptosis in cardiomyocytes.METHODS:Rat cardiomyocytes were cultured in vitro and divided into control group and Hcy group.After 48 h of intervention,the expression levels of cuproptosis-related proteins,ferre-doxin 1(FDX1)and heat shock protein 70(HSP70),were detected by Western blot and immunofluorescence staining.The oxidative stress state of cardiomyocytes was assessed using fluorescence staining,and the intracellular Cu2+levels were measured using a copper ion assay kit.Furthermore,the impact of Hcy on the expression of cuproptosis-related proteins in cardiomyocytes was analyzed following overexpression of lncSLC25a6.RESULTS:Compared with the control group,80 μmol/L Hcy significantly accelerated cardiomyocyte damage,with a notable underexpression of lncSLC25a6(P<0.05).Western blot results indicated that,compared with the control group,the expression level of FDX1 in the Hcy intervention group was significantly reduced(P<0.05),while the expression level of HSP70 was significantly elevated(P<0.05),and the expression level of copper ions in cardiomyocytes of the Hcy group was increased(P<0.05).Immunofluorescence staining showed a significant reduction in FDX1 fluorescence intensity and a significant increase in HSP70 fluorescence in-tensity in the Hcy group.Further overexpression of lncSLC25a6 significantly mitigated Hcy-induced cuproptosis in cardio-myocytes,resulting in elevated expression of FDX1 and reduced expression of HSP70(P<0.05).Pearson correlation analysis demonstrated that the expression level of lncSLC25a6 was negatively correlated with FDX1 protein expression(r=-0.676,P=0.046)and positively correlated with HSP70 expression(r=0.680,P=0.044).CONCLUSION:lnc-SLC25a6 significantly mitigates Hcy-induced cuproptosis in cardiomyocytes,positioning it as a potential therapeutic target for managing Hcy-induced cardiac injury.
6.Analysis on the feasibility of short collection time of step by step mode of digital PET/CT
Jianqiang LIU ; Xiaopin LI ; Jiajun LI ; Hongyang JIAO ; Fuhe LIU ; Jingzhe LIU
China Medical Equipment 2024;21(8):7-12
Objective:To analyze the effect of short collection time of digital positron emission tomography/computed tomography(PET/CT)on image quality,detected lesion and semi-quantitative parameters,and explore the feasibility of clinical application of short collection time of step by step collection mode.Methods:The digital PET/CT images of 63 subjects were retrospectively analyzed.All of them adopted PET/CT scan to conduct PET/CT examination for whole body according to the collection mode of step by step type and list-mode scan protocol.The reconstruction was implemented according to the different collection time of each bed,and they were divided into 90 s/bed group,60 s/bed group,45 s/bed group and 30 s/bed group.The 90s/bed was used as control group to compare the detection rate of the small lesion of each group("the shorter diameter of smaller radioactive concentration lesion of PET image and the lesion of CT image"was less than 0.5 cm)in PET images,and to measure the objective data of original tumor lesion,and to compare the maximum value(SUVmax),peak value(SUVpeak)and mean value(SUVmean)of standardized uptake value(SUV),noise(SD),metabolic tumor volume(MTV)and total lesion glycolysis(TLG).The signal-noise ratio(SNR),contrast noise ratio(CNR)and target-to-background ratio(TBR)were calculated and compared.The one-way analysis of variance(ANOVA test)was adopted to conduct comparison of inter groups for the image data of 4 groups.The Bland-Altman consistency test was adopted to analyze semi-quantitative parameters of intra group.The Likert 5 scores method was adopted to conduct subject evaluation for the quality of images.The Kappa test was adopted to evaluate the consistency of images.Results:The subjective scores of image quality of 4 groups were all≥3 points,and the consistency of image quality among the four groups was very well(Kappa=0.8,P<0.05).Compared with 90s/bed group,the detection rate of small lesions of other three groups were 100%.The differences of SUVmax,SUVpeak,SUVmean,SD,TLG and MTV of semi-quantitative indicators among 4 groups were not significant(P>0.05).There were significant differences in CNR,SNR and TBR of objective indicators of 18F-FDG PET image quality among 4 groups(F=31.741,34.199,12.021,P<0.001),respectively.There were no significant differences in noise between 60s/bed group and 90s/bed group,and between 45s/bed group and 90s/bed group,respectively.The noise of image of 30s/bed group appeared increase.The Bland-Altman consistent analysis of intra group basically was within 95%confidence interval(CI).Conclusions:The short collection time of step by step mode of digital PET/CT has very good clinical application.
7.Clinical application value of single pass scanning using multi-slice spiral CT for polytrauma patients
Gang WANG ; Ying ZHANG ; Teng TANG ; Hongyang DAI ; Nan WANG ; Mimi QIU ; Hongwei XU ; Fangfang XIA ; Xinglin XIANG ; Jian LI
Chinese Journal of Radiological Medicine and Protection 2024;44(8):693-698
Objective:To explore the clinical application value of single pass scanning using muti-slice spiral CT for polytrauma patients.Methods:Totally 60 polytrauma patients treated from January to November in 2023 were randomly enrolled in this study. They were categorized into an experimental group and a control group using a random number table, with 30 patients in each group. The patients in the experimental group underwent single pass scaning for the head, neck, chest, and abdomen, whereas those in the control group receiving separate scanning for various parts. Then, the noise, signal-to-noise ratio (SNR), and contrast-to-noise (CNR) of the CT images of both groups were recorded. Furthermore, the objective and subjective evaluation, volume CT dose index (CTDI vol), effective dose ( E), scanning time, and scan ranges of the images were compared between both groups. Results:Compared to the control group, the test group exhibited lower SNR of the head ( t = -5.47, P < 0.05) and higher SNR and CNR of the chest scans ( t = -5.95, -6.15, P < 0.05). Furthermore, the test group demonstrated decreased ED, CTDIvol, scanning time, and scan range, which dropped from 18.53 mSv to 13.81 mSv ( t = 3.29, P < 0.001), from 15.77 mGy to 10.59 mGy ( t = 4.48, P< 0.001), from 31.68 s to 10.97 s ( t = 6.95, P < 0.001), and from 64.92 cm to 45.21 cm ( t = 9.05, P < 0.001), respectively compared to the control group. Conclusions:Single pass CT scanning can reduce E, scanning time, and scan range in the treatment of polytrauma patients while ensuring the high quality of CT images, thus warranting wide clinical applications.
8.Biocontrol Effect and Culture Condition Optimization of Trichoderma citrinoviride Strain Isolated from Rhizosphere of Atractylodes lancea
Kuo LI ; Sheng WANG ; Hongyang WANG ; Zheng PENG ; Xiuzhi GUO ; Tielin WANG ; Yuefeng WANG ; Lanping GUO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):192-199
ObjectiveRoot rot is one of the most serious diseases in the cultivation and production of Atractylodes lancea. Trichoderma spp. are effective in the biocontrol of root rot without causing environmental pollution. This study aims to isolate and study a Trichoderma strain capable preventing and controlling root rot from the rhizosphere of A. lancea and to solve the problem of disease prevention and control in the planting and production of A. lancea. MethodTrichoderma T2204 was isolated by the dilution-coating method and identified by ITS sequencing. The inhibitory activities of T2204 and its volatiles against two pathogenic fungal strains were examined by dual-culture and co-culture experiments. The biocontrol potential of T2204 on root rot of A. lancea and the effect of T2204 on the accumulation of medicinal compounds in the rhizosphere of A. lancea were investigated by pot experiments and GC-MS, respectively. In addition, the optimal medium, photoperiod, temperature, pH, and carbon and nitrogen sources for the culture of T2204 were explored. ResultThe Trichoderma isolate T2204 was identified as T. citrinoviride and had direct inhibitory effects on two highly pathogenic strains causing root rot. In the dual-culture experiments with the two pathogenic strains, T2204 showcased the inhibition rates of 77.90% and 76.80%, respectively. In the co-culture experiments with the two pathogenic strains, the volatile organic compounds produced by T2204 showed the inhibition rates of 57.11% and 81.11%, respectively. The pot experiments showed that the survival rate of A. lancea seedlings infected by root rot reached 100% after inoculation with T2204 and was only 50% in the case without inoculation of T2204. After 150 days of cultivation, the dry weight and atractylodin content of the rhizome of A. lancea plants treated with T2204 increased by 32% (P<0.05) and 11%, respectively, compared with the untreated group. The optimal conditions for the growth of T2204 were PDA or PSA medium, photoperiod of 12 h dark/12 h light, 25-30 °C, pH 5-6, carbon sources of glucose, D-fructose, soluble starch, and maltose, and the nitrogen sources of ammonium sulfate and ammonium dihydrogen phosphate. The optimal conditions for the sporulation of T2204 were PSA or CMA medium, photoperiod of 12 h dark/12 h light, 20-30 °C, pH 8, carbon source of sucrose, and nitrogen source of sodium nitrate. ConclusionT2204 could improve the growth and root rot resistance of A. lancea and promote the accumulation of medicinal compounds. The findings laid a foundation for the industrialized production and application of T2204 in the production of A. lancea in the future.
9.Spatial and temporal analysis of MDR-TB epidemic based on SaTScan in Nanning in 2017 - 2021
Shu LI ; Yuan QIU ; Bo HE ; Hongyang TANG ; Chengyuan LEI ; Jiayun HUANG
Journal of Public Health and Preventive Medicine 2024;35(2):17-20
Objectives To analyze the spatial and temporal aggregation of multidrug resistant pulmonary tuberculosis (MDR-TB) incidence in Nanning at the township / street scale from 2017 to 2021, to explore the spatial and temporal characteristics of the spread of MDR-TB in Nanning, and to provide a scientific reference basis for the health administrative departments to achieve the precise implementation of MDR-TB prevention and control. Methods Based on the data of MDR-TB cases in Nanning from 2017 to 2021, the spatial-temporal scanning analysis software SaTScan v9.7 was used to retrospectively detect and analyze the areas where MDR-TB cases gathered. Results Through simple spatial scanning analysis, it was found that there were three first-class aggregation areas (the aggregation center was Fujiayuan Street, Jiangnan District, 2017, Xinyang Street, Xixiangtang District, 2019, and Zhonghe Town, Yongning District, 2020), and one second-class aggregation area (the aggregation center was Jinchai Town, Mashan County, 2020). Simple time scanning showed that the clustering occurred from May 2019 to December 2020. Temporal and spatial aggregation analysis showed that Xinyang Street in Xixiangtang District was the center of the first-class aggregation area, Zhonghe Town in Yongning District was the center of the second-class aggregation area, and Jinchai Town in Mashan County was the center of the third-class aggregation area. Conclusion The multidrug resistant pulmonary tuberculosis epidemic in Nanning is distributed in an aggregated manner, especially in Xinyang Street, Xixiangtang District, which has the highest spatial and temporal aggregation. It is necessary to focus on and take regional prevention and control measures to control the epidemic.
10.Distribution characteristics and correlation analysis of GJB2 variation in patients with auditory neuropathy.
Yiming LI ; Hongyang WANG ; Danyang LI ; Qiuju WANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2024;38(1):23-29
Objective:To elucidate the correlation between the GJB2 gene and auditory neuropathy, aiming to provide valuable insights for genetic counseling of affected individuals and their families. Methods:The general information, audiological data(including pure tone audiometry, distorted otoacoustic emission, auditory brainstem response, electrocochlography), imaging data and genetic test data of 117 auditory neuropathy patients, and the patients with GJB2 gene mutation were screened out for the correlation analysis of auditory neuropathy. Results:Total of 16 patients were found to have GJB2 gene mutations, all of which were pathogenic or likely pathogenic.was Among them, one patient had compound heterozygous variants GJB2[c. 427C>T][c. 358_360del], exhibiting total deafness. One was GJB2[c. 299_300delAT][c. 35_36insG]compound heterozygous variants, the audiological findings were severe hearing loss.The remaining 14 patients with GJB2 gene variants exhibited typical auditory neuropathy. Conclusion:In this study, the relationship between GJB2 gene and auditory neuropathy was preliminarily analyzed,and explained the possible pathogenic mechanism of GJB2 gene variants that may be related to auditory neuropathy.
Humans
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Connexins/genetics*
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Connexin 26/genetics*
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Hearing Loss, Central/genetics*
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Deafness/genetics*
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Mutation


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