1.Characteristic volatile organic compounds in exhaled breath of coal workers' pneumoconiosis patients by thermal desorption gas chromatography-mass spectrometry
Yazhen HE ; Chunguang DING ; Junyun WANG ; Yuzhen FENG ; Fangda PENG ; Gaisheng LIU ; Fan YANG ; Chunmin ZHANG ; Rui GAO ; Qingyu MENG ; Zhijun WU ; Jingguang FAN
Journal of Environmental and Occupational Medicine 2025;42(5):571-577
Background Coal workers' pneumoconiosis is a serious occupational disease in China. Exhaled volatile organic compounds (VOCs) can serve as the "breath fingerprint" of internal pathological processes, which provides a theoretical basis for exhaled VOCs to be used as potential non-invasive biomarkers for early diagnosis of coal workers' pneumoconiosis. Objective To screen out the characteristic VOCs and important characteristic VOCs of exhaled air in patients with coal workers' pneumoconiosis, and to explore the potential of these VOCs as biomarkers for early non-invasive diagnosis of the disease. Methods In this study, 27 VOCs in the exhaled breath of 22 patients with stage I coal workers' pneumoconiosis, 77 workers exposed to dust, and 92 healthy controls were quantitatively detected by thermal desorption gas chromatography-mass spectrometry (TD-GC-MS). Substances with P<0.05 in univariate analysis and variable importance projection (VIP) >1 in supervised orthogonal partial least squares discriminant analysis (OPLS-DA) model were selected as the characteristic VOCs for early diagnosis of coal workers' pneumoconiosis. Age was included in the LASSO regression model as a covariate to screen out important characteristic VOCs, and the diagnostic performance was evaluated by receiver operating characteristic (ROC) curve. Spearman correlation was further used to explore the correlation between important characteristic VOCs and clinical lung function indicators. Results Through univariate analysis and OPLS-DA modeling, 8 VOCs were selected, including 2-methylpentane, 3-methylpentane, n-hexane, methylcyclopentane, n-heptane, methylcyclohexane, 4-methyl-2-pentanone, and 2-hexanone, in exhaled breath of patients with coal workers' pneumoconiosis. The concentrations of 4 VOCs, including 3-methylpentane, n-hexane, 4-methyl-2-pentanone, and 2-hexanone, showed a decreasing trend with the increase of dust exposure years. By LASSO regression, the important characteristic VOCs of the coal workers' pneumoconiosis group and the dust exposure group were n-hexane, methylcyclohexane and 4-methyl-2-pentanone, and the important characteristic VOCs of the coal workers' pneumoconiosis group and the healthy group were 2-methyl-pentane and 4-methyl-2-pentanone. The ROC analysis showed that the area under the curve (AUC) of n-hexane, methylcyclohexane, and 4-methyl-2-pentanone were 0.969, 0.909, and 0.956, respectively, and the AUC of combined diagnosis was 0.988 and its Youden index was 0.961, suggesting that these results can serve as a valuable reference for further research on early diagnosis. The Correlation analysis found that there was a positive correlation between n-hexane and lung function indicators in the important characteristic VOCs, indicating that it could indirectly reflect the obstruction of lung function ventilation, further proving that important characteristic VOCs have the potential to monitor lung function decline. Conclusion Three important characteristic VOCs selected in this study have the potential to be used as non-invasive biomarkers for early diagnosis and disease monitoring of coal workers' pneumoconiosis, and are worthy of further study and verification.
2.Determination of 27 volatile organic compounds in exhaled breath by thermal desorption-gas chromatography-mass spectrometry
Rui GAO ; Fangda PENG ; Yazhen HE ; Jie FENG ; Wubin LIU ; Lijia YU ; Qingsong CHEN ; Chunguang DING
Journal of Environmental and Occupational Medicine 2024;41(1):96-102
Background Volatile organic compounds (VOCs) in exhaled breath are closely associated with respiratory diseases and are linked to various metabolic reactions in the human body. A quantitative analytical method can provide technical support for studying VOCs related to various diseases. Objective To establish a thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) method for the determination of 27 VOCs in exhaled breath. Methods VOCs in exhaled breath were collected using a Bio-VOC sampler and enriched with Tenax TA thermal desorption tubes before TD-GC-MS analysis. Standards were collected using thermal desorption tubes and optimized for thermal desorption conditions as well as chromatographic and mass spectrometric conditions: The separation of the 27 VOCs was achieved by an optimized temperature program, the improvement of sensitivity by optimizing quantitative ions, and the increase of VOCs desorption efficiency by optimizing thermal desorption time and temperature. Limit of detection, limit of quantification, accuracy, precision, and stability of the proposed method were investigated by spiking with a blank gas bag, and exhaled breath samples from 20 healthy individuals were collected for an application study of the proposed method. Results The thermal desorption temperature was 280 ℃, and desorption time was 6 min. A VF-624ms chromatographic column was selected for the separation of target substances. The initial temperature of heating program was 35 ℃, maintained for 1 min, and then increased to 100 ℃ at a heating rate of 3 ℃·min−1 for 1 min, followed by increasing to 210 ℃ at a heating rate of 28 ℃·min−1 for 5 min. A quantitative analysis was conducted with a single ion monitoring (SIM) mode. Under these conditions, the 27 VOCs showed good linear relationships in their respective concentration ranges and the correlation coefficients were higher than 0.9990. The limits of detection of the method were in the range of 0.01-0.13 nmol·mol−1, the limits of quantification were in the range of 0.02-0.44 nmol·mol−1, and the spiked recoveries were in the range of 80.1%-120.5%, with intra-batch and inter-batch precision ≤ 18.8% and 17.9% respectively. All substances can be stored at room temperature (23-28 °C) for 7 d and at 4 °C for 14 d. The proposed method was applied to exhaled breath samples from 20 subjects with detection rates≥ 80% (except for trans-2-pentene and decane) and a concentration range of 0.00-465.50 nmol·mol−1. Conclusion The established TD-GC-MS method for quantification of VOCs in exhaled breath is characterized by high sensitivity and good accuracy, and is suitable for quantitative determination of VOCs in exhaled breath, which can provide technical support for the study of exhaled breath VOCs.
3.Ku70 Functions as an RNA Helicase to Regulate miR-124 Maturation and Neuronal Cell Differentiation
Ai-Xue HUANG ; Rui-Ting LI ; Yue-Chao ZHAO ; Jie LI ; Hui LI ; Xue-Feng DING ; Lin WANG ; Can XIAO ; Xue-Mei LIU ; Cheng-Feng QIN ; Ning-Sheng SHAO
Progress in Biochemistry and Biophysics 2024;51(6):1418-1433
ObjectiveHuman Ku70 protein mainly involves the non-homologous end joining (NHEJ) repair of double-stranded DNA breaks (DSB) through its DNA-binding properties, and it is recently reported having an RNA-binding ability. This paper is to explore whether Ku70 has RNA helicase activity and affects miRNA maturation. MethodsRNAs bound to Ku protein were analyzed by RNA immunoprecipitation sequencing (RIP-seq) and bioinfomatic anaylsis. The expression relationship between Ku protein and miRNAs was verified by Western blot (WB) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays. Binding ability of Ku protein to the RNAs was tested by biolayer interferometry (BLI) assay. RNA helicase activity of Ku protein was identified with EMSA assay. The effect of Ku70 regulated miR-124 on neuronal differentiation was performed by morphology analysis, WB and immunofluorescence assays with or without Zika virus (ZIKV) infection. ResultsWe revealed that the Ku70 protein had RNA helicase activity and affected miRNA maturation. Deficiency of Ku70 led to the up-regulation of a large number of mature miRNAs, especially neuronal specific miRNAs like miR-124. The knockdown of Ku70 promoted neuronal differentiation in human neural progenitor cells (hNPCs) and SH-SY5Y cells by boosting miR-124 maturation. Importantly, ZIKV infection reduced the expression of Ku70 whereas increased expression of miR-124 in hNPCs, and led to morphologically neuronal differentiation. ConclusionOur study revealed a novel function of Ku70 as an RNA helicase and regulating miRNA maturation. The reduced expression of Ku70 with ZIKV infection increased the expression of miR-124 and led to the premature differentiation of embryonic neural progenitor cells, which might be one of the causes of microcephaly.
4.Bibliometric study on medical Chinese core journals literature in inner mongolia autonomous region from 2018 to 2022
Yingjie TIAN ; Rui FENG ; Shangyin MA ; Haitao DING ; Zhanguo WANG
Modern Hospital 2024;24(7):1134-1136,1141
Objective To analyze the quantity,quality,and research layout of medical scientific papers(in Chinese)published by various regions,hospitals,and institutions in Inner Mongolia Autonomous Region from 2018 to 2022,in order to understand the overall scientific and technological level of our region,providing an overview of support for medical scientific research in the entire region and provi-ding references for the rational layout of medical scientific and technological development.Methods From July to August 2023,Chinese National Knowledge Infrastructure(CNKI)was used as the data source to systematically retrieve Chinese medical literature related to Inner Mongolia Autonomous Region from 2018 to 2022.Bibliometric methods were applied to analyze the quantity and quality of literature,as well as regional and institutional distribution.Results From 2018 to 2022,there were 5 273 articles published in core journals in Inner Mongo-lia,accounting for 30.96%of the total.Among them,1095 articles(20.77%)were indexed in CSCD,1 567 articles(29.72%)in Peking University Core Journals,and5164 articles(97.93%)in Science and Technology Core Journals.The journal with the highest publication volume among the top10 core journals was"Journal of Inner Mongolia Medical University,"accounting for20.77%.The top three regions in terms of total output of core journal articles were Hohhot,Baotou,and Chifeng,while Xing'an League and Alxa League had very few pub-lications.Among different levels of hospitals,tertiary hospitals had much higher publication volumes than primary and secondary hospitals.Among different types of institutions,medical institutions had higher publication volumes than higher education institutions.The top three in-stitutions in terms of publication volumes in core journals were Affiliated Hospital of Inner Mongolia Medical University,Inner Mongolia Med-ical University,and Inner Mongolia People's Hospital.Conclusion The number of core journals in Inner Mongolia needs to be increased,and the quantity of research output varies among regions.It is necessary to strengthen scientific research management systems,improve the level of research achievements,and promote scientific and technological innovation in Inner Mongolia.
5.Clinical value of high frequency ultrasound in the diagnosis of hip joint involvement in patients with ankylosing spondylitis
Guo-Yan XUE ; Ying-Qi LI ; Rui WANG ; Wen-Xing WANG ; Zhuang DONG ; Jian-Feng DING
China Journal of Orthopaedics and Traumatology 2024;37(4):374-380
Objective To analyze the correlation between hip joint musculoskeletal ultrasound score and ankylosing spondylitis(AS)disease activity,as well as to investigate the value of high frequency ultrasound in the assessment of hip joint involvement in AS.Methods The clinical data of 244 patients with AS who were treated in the rheumatology department of from March 2019 to March 2022 were retrospectively analyzed.Among them,there 174 males and 70 females,aged from 19 to 58 years old with an average of(34.22±9.49)years old;the disease duration of AS patients ranged from 8 months to 26 years,with an average of(13.68±4.04)years.The 244 patients were divided into disease group(83 cases)and control group(161 cases)based in the presence of hip joint involuement.According to the the disease activity,patients in the disease group were further categorezed into active phase(45 cases)and stable phase(38 cases).The ultrasound scores of patients in the active and stable phases of the disease group and the control group were compared.Relevant factors of hip joint involvement in AS patients were analyzed,and analyze the correlation between ultrasound score and Bath ankylosing spondylitis disease activity score index(BASD AI),Bath ankylosing spondylitis functional index(BASFI),visual analogue score of pain(VAS),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),and the correlation between hip joint capsule score and tendon attach-ment end score and BASDAI,BASFI,VAS,CRP and ESR.Results The hip joint capsule score(3.06±1.12),femoral head score(1.45±0.43),tendon attachment end score(3.28±1.30)and total ultrasound score(6.65±2.31)of the disease group were higher than those of the control group(1.51±0.48)、(0.66±0.27)、(1.61±0.53)、(3.81±1.44)scores(P<0.05).Multifactor Logstic re-gression analysis showed that the course of disease,hip joint capsule score and total ultrasound score were independent risk factors for hip involvement in AS patients.The hip capsule score(3.65±1.22)and total ultrasound score(8.28±2.33)in the ac-tive phase of the disease group were higher than those in the stable phase(2.48±1.04)、(6.82±1.96)scores(P<0.05).The hip joint capsule score and total ultrasonic score of AS patients were positively correlated with BASDAI,BASFI,VAS,CRP,and ESR(P<0.05,P<0.01).The score of tendon attachment end was positively correlated with CRP(P<0.05).The score of joint capsule effusion in AS patients was positively correlated with BASDAI,BASFI and VAS(P<0.05,P<0.01).The synovial blood flow score was positively correlated with BASDAI,VAS,CRP and ESR(P<0.05,P<0.01).The synovial thickening score was positively correlated with BASDAI,BASFI,VAS,CRP and ESR(P<0.05,P<0.01).There was no correlation between the score of tendon attachment end and BASDAI,BASFI,VAS,CRP and ESR.Conclusion There is a correlation between hip joint ul-trasonic score of hip joint and clinical indexes in AS patients.Hip joint capsule score and total ultrasonic score were indepen-dent risk factors for hip involvement in AS patients.High frequency ultrasound exhibits clinical value in the diagnosis of hip joint involvement in AS patients.
6.Implantation of Adipose-Derived Mesenchymal Stromal Cells (ADSCs)-Lining Prosthetic Graft Promotes Vascular Regeneration in Monkeys and Pigs
Xiao ZUO ; Pengfei HAN ; Ding YUAN ; Ying XIAO ; Yushi HUANG ; Rui LI ; Xia JIANG ; Li FENG ; Yijun LI ; Yaya ZHANG ; Ping ZHU ; Hongge WANG ; Ning WANG ; Y. James KANG
Tissue Engineering and Regenerative Medicine 2024;21(4):641-651
BACKGROUND:
Current replacement procedures for stenosis or occluded arteries using prosthetic grafts have serious limitations in clinical applications, particularly, endothelialization of the luminal surface is a long-standing unresolved problem.METHOD: We produced a cell-based hybrid vascular graft using a bioink engulfing adipose-derived mesenchymal stromal cells (ADSCs) and a 3D bioprinting process lining the ADSCs on the luminal surface of GORE-Tex grafts. The hybrid graft was implanted as an interposition conduit to replace a 3-cm-long segment of the infrarenal abdominal aorta in Rhesus monkeys.
RESULTS:
Complete endothelium layer and smooth muscle layer were fully developed within 21 days post-implantation, along with normalized collagen deposition and crosslinking in the regenerated vasculature in all monkeys. The regenerated blood vessels showed normal functionality for the longest observation of more than 1650 days. The same procedure was also conducted in miniature pigs for the interposition replacement of a 10-cm-long right iliac artery and showed the same long-term effective and safe outcome.
CONCLUSION
This cell-based vascular graft is ready to undergo clinical trials for human patients.
7. Mechanism of Transcription Factor SP1 Regulates Expression of Tripartite Motif-containing Protein 7 in Hepatocellular Carcinoma
Chang-Yu FAN ; Han DING ; Yi-Ran XU ; Rui XIAN ; Feng-Juan YAN
Chinese Journal of Biochemistry and Molecular Biology 2023;39(9):1314-1321
Tripartite motif containing protein 7 (TRIM7), as a member of the E3 ubiquitin ligase TRIM family, plays an important regulatory role in immune regulation, metabolism and other physiological processes. The aberrant expression of TRIM7 is closely related to the development and progression of hepatocellular carcinoma (HCC) and it shows a complex regulatory role. However, the regulatory mechanism for the expression of TRIM7 in HCC remains unknown. In this study, multiple online databases were used to analyze the expression of TRIM7 in HCC and data indicated that TRIM7 expression was upregulated in HCC and correlated to poor prognosis. Subsequently, the transcription factor binding sites in the TRIM7 promoter region were analyzed using UCSC and JASPAR databases, and the results showed that TRIM7 promoter contains four SP1 binding sites. In this work, we demonstrated that SP1 could directly bind to its binding sites in TRIM7 promoter and positively regulate the transcriptional activity driven by the TRIM7 promoter using dual luciferase reporter experiments and the ChIP-PCR method. Moreover, our results also showed SP1 overexpression upregulated the expression of TRIM7 at both mRNA and protein levels (P<0. 01),and SP1 inhibitor, mithramycin A, could reverse the activated effect of SP1 on TRIM7 expression (P<0. 01). In conclusion, this study preliminarily reveals the regulatory mechanism of TRIM7 upregulation in HCC, which provides an important theoretical basis for further study of the gene function, early diagnosis and targeted therapy.
8.Analysis of Predictive Factors for Efficacy of Yiqi Yangxue Prescription Combined with Western Medicine in Treatment of Aplastic Anemia in Non-elderly Adults
Chaochang ZHANG ; Rui LI ; Jian LIU ; Yubin DING ; Jinhuan WANG ; Qifeng LIU ; Ruirong XU ; Haixia WANG ; Haixia DI ; Antao SUN ; Yonggang XU ; Xiupeng YANG ; Feng LIU ; Rou MA ; Xudong TANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(20):88-98
ObjectiveTo explore the predictive factors for the efficacy of Yiqi Yangxue prescription combined with western medicine in treating aplastic anemia (AA) in non-elderly adults, so as to provide a reference for predicting the prognosis of this therapy. MethodA retrospective study was conducted with the clinical data of non-elderly adult AA patients who visited 19 hospitals including Xiyuan Hospital of the China Academy of Chinese Medical Sciences from September 2018 to March 2021 and were treated with Yiqi Yangxue Prescription combined with western medicine. According to the efficacy evaluation results at the 6th month of treatment, the patients were assigned into effective and ineffective groups. The two groups were compared in terms of the gender, age, disease classification [non-severe aplastic anemia (NSAA)/severe aplastic anemia (SAA)], course of disease, family history, complications, history of drug allergy, baseline blood routine examination [hemoglobin (HGB), white blood cell (WBC), neutrophil (ANC), platelet (PLT), and reticulocyte (Ret)], T lymphocyte subsets, degree of proliferation of nucleated cells in bone marrow, and expression of T-bet and GATA-3. ResultA total of 101 non-elderly adult AA patients were enrolled in this study, including 81 in the effective group and 20 in the ineffective group. The effective group had a higher proportion of the patients without a history of drug allergy than the ineffective group (P<0.05). The body height, body weight, gender, age, disease classification, course of disease, family history, and complications showed no significant differences between two groups. The effective group had higher levels of ANC and PLT before treatment (P<0.05) and higher proportion of patients with ANC≥1.6×109/L and PLT≥25×109/L (P<0.05, P<0.01) than the ineffective group. The baseline levels of WBC, HGB, and Ret showed no significant statistical differences between two groups. The levels of CD3+HLA-DR+T cells in the effective group before treatment was higher than that in the ineffective group (P<0.05). The levels of CD3+CD19-T cells, CD4+T cells, CD8+T cells, Th1 cells, Th2 cells, and CD3+CD25+T cells showed no significant statistical differences between two groups before treatment. The proportion of patients with active bone marrow nucleated cells proliferation in the effective group before treatment were significantly higher than that in the ineffective group, while the proportion of patients with reduced or extremely reduced proliferation were significantly lower than that in the ineffective group (P<0.05). The expression levels of T-bet and GATA-3 genes had no significant differences between two groups before treatment. The multivariate binary logistic regression analysis showed that the ANC level before treatment and history of drug allergy were independent influencing factors for efficacy (P<0.05, P<0.01), while other indicators were not influencing factors for efficacy. The receiver operating characteristic (ROC) curve was applied to analyze the predictive value of the ANC level before treatment in the treatment of AA in non-elderly adults with Yiqi Yangxue prescription combined with western medicine. The area under the curve was 0.679 (P<0.05), with the critical value of 1.595×109/L, the sensitivity of 0.42, and the specificity of 0.95. ConclusionThe history of drug allergy, pre-treatment ANC, PLT, CD3+HLA-DR+ T cell levels, and proliferation of nucleated cells in bone marrow before treatment are predictive factors for the efficacy of Yiqi Yangxue prescription combined with western medicine in treating AA in non-elderly adults. This therapy tends to be more effective for the patients with no history of drug allergy, higher ANC and PLT levels before treatment, especially those with ANC≥1.6×109/L, PLT≥25×109/L, and higher CD3+ HLA-DR+T cell levels and the more active proliferation of nucleated cells in bone marrow before treatment.
9.Evaluation of macular function changes in glaucoma using MAIA microperimetry
Hong JU ; Rui HAO ; Yun GU ; Zhao-Feng DING
International Eye Science 2023;23(12):1950-1953
AIM: To observe the characteristics of microperimetry in patients with glaucoma, and investigate the correlation between microperimetry and best corrected visual acuity(BCVA), standard automatic perimetry and optical coherence tomography(OCT)index.METHODS: This case-control study included 45 patients(76 eyes)with glaucoma(glaucoma group), among which 15 patients(25 eyes)with primary open-angle glaucoma and 30 patients(51 eyes)with chronic angle-closure glaucoma, and 40 healthy individuals(76 eyes)were included in the control group. MAIA microperimetry, Humphrey perimetry, and BCVA tests were performed in all examined eyes. Correlation analysis was performed using the structural indices measured by OCT.RESULTS: The mean sensitivity(MS)in macular area measured by microperimetry and the ganglion cell complex(GCC)and retinal nerve fiber layer(RNFL)thickness measured by OCT were decreased in glaucoma patients when compared to the control group. Additionally, the mean defect(MD)measured by Humphrey perimetry(10-2 visual field test), focal loss volume(FLV), global loss volume(GLV)measured by OCT, and 63% bivariate contour ellipse area(BCEA )measured by MAIA microperimetry were higher than those of the control group. The MS was negatively correlated with MD, FLV, GLV and BCVA(LogMAR)in the glaucoma group(rs=-0.839, -0.665, -0.530, and -0.424, all P<0.01). In contrast, MS was positively correlated with GCC and RNFL in the glaucoma group(rs=0.437, 0.500, all P<0.01). MAIA microperimetry had a shorter detection time. Receiver operating characteristic(ROC)curve analysis showed that MAIA microperimetry had moderate accuracy for the diagnosis of glaucoma.CONCLUSIONS: MAIA microperimetry has high sensitivity and can detect retinal sensitivity reduction in areas of structural damage tested by OCT. The microperimetry values were correlated with BCVA, standard automatic perimetry parameters, and OCT parameters. MAIA microperimetry combined with OCT will increase the early diagnosis rate of glaucoma.
10.Construction and optimization of a novel plasma membrane model for membrane affinity assay based on polydiacetylene vesicles
Chai-chai NIE ; Rui-ting DONG ; Yu-tong WU ; Jing-bo WU ; Sheng ZHANG ; Feng ZHENG ; Ya DING
Acta Pharmaceutica Sinica 2023;58(8):2503-2511
Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.

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