1.Simultaneous determination of four thiol derivatives in workplace air by gas chromatography
Ruibo MENG ; Jing YUAN ; Jiawen HU ; Jiaheng HE ; Jingjing QIU ; Zuokan LIN ; Ziqun ZHANG ; Weifeng RONG ; Banghua WU
China Occupational Medicine 2025;52(2):188-192
Objective To establish a method for simultaneous determination of four high-molecular-weight thiol derivatives (TDs) in workplace air by gas chromatography. Methods The four kinds of vapor-phase macromolecular TDs (1-pentanethiol, 1-hexanethiol, 1-benzyl mercaptan, and n-octanethiol) in the workplace air were collected using the GDH-1 air sampling tubes, desorbed with anhydrous ethanol, separated on a DB-FFAP capillary column, and determined by flame ionization detector. Results The quantitation range of the four TDs was 0.30-207.37 mg/L, with the correlation coefficients greater than 0.999 00. The minimum detection mass concentrations and minimum quantitation mass concentrations were 0.18-0.32 and 0.60-1.05 mg/m3, respectively (both calculated based on the 1.5 L sample and 3.0 mL desorption solvent). The mean desorption efficiencies ranged from 87.07% to 103.59%. The within-run and between-run relative standard deviations were 1.92%-8.22% and 1.89%-8.45%, respectively. The samples can be stored at room temperature or 4 ℃ for three days and up to 7 days at -18 ℃. Conclusion This method is suitable for the simultaneous determination of four vapor-phase TDs in workplace air.
2.Erythropoietin-overexpressed umbilical cord mesenchymal stem cells inhibit neuroapoptosis in ischemic-hypoxic SH-SY5Y and its mechanism
Ruibo LI ; Ning KONG ; Lei SUN ; Baodong MA ; Ranran JIN ; Wenjin ZHANG ; Han YUE ; Hui ZHANG
Chinese Journal of Tissue Engineering Research 2024;28(31):4937-4944
BACKGROUND:Previous studies have successfully constructed erythropoietin-overexpressed umbilical cord mesenchymal stem cells.It was found that the apoptosis of ischemic and hypoxic human neuroblastoma cell line(SH-SY5Y)was significantly reduced by erythropoietin-overexpressed umbilical cord mesenchymal stem cells. OBJECTIVE:To explore the possible neuroprotective mechanisms of erythropoietin-overexpressed umbilical cord mesenchymal stem cells against ischemic-hypoxic SH-SY5Y and their associated epigenetic mechanisms. METHODS:Oxygen-glucose deprivation was applied to ischemia-hypoxia-induced SH-SY5Y cell injury,and multifactorial assays were applied to detect the expression levels of inflammatory factors in the cells before and after hypoxia and co-culture,respectively,with mesenchymal stem cells,as well as lentiviral-transfected null-loaded plasmids of the negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression levels of supernatant inflammatory factors were detected by multifactor assay after co-culture.Proteomics was used to detect the differentially expressed proteins of negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.Cleavage under targets and tagmentation sequencing was applied to detect genomic H3K4me2 modification,and joint analysis was conducted with RNA-sequencing.Lentiviral vector infection was applied to construct the stable knockdown of REST in SH-SY5Y cells.qRT-PCR and western blot assay were performed to detect the expression level of REST.The apoptosis was detected by flow cytometry after co-culture of oxygen-glucose deprivation treatment with erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression difference of H3K36me3 group proteins was detected by western blot assay,and transcriptome sequencing was performed to analyze the differentially expressed genes. RESULTS AND CONCLUSION:(1)Compared with the control group,monocyte chemotactic protein 1,interleukin-6,interleukin-18,and interleukin-1 beta,interferon α2,and interleukin-23 levels significantly increased in the cerebrospinal fluid supernatant of patients with ischemic-hypoxic encephalopathy(P<0.01).(2)After co-culturing SH-SY5Y cells with erythropoietin-overexpressed umbilical cord mesenchymal stem cells under ischemia and hypoxia,the expression levels of monocyte chemotactic protein 1 and interleukin-6 were significantly reduced.(3)Analysis of protein network interactions revealed significant downregulation of monocyte chemotactic protein 1,interleukin-6 related regulatory proteins CXCL1 and BGN.(4)Transcriptome sequencing analysis found that pro-inflammatory genes were down-regulated,and functional enrichment of histone modifications,and the expression of transcription factors REST and TET3 significantly up-regulated in the erythropoietin-overexpressed umbilical cord mesenchymal stem cell group compared with the negative control mesenchymal stem cell group.(5)Combined analysis of transcriptome sequencing and cleavage under targets and tagmentation revealed changes in epigenetic levels as well as significant activation of the promoter regions of transcription factors REST and TET3.(6)Stable knockdown REST in SH-SY5Y cells was successfully constructed;the transcript levels of REST mRNA and protein expression were both decreased.(7)After the REST knockdown SH-SY5Y cells were co-cultured with erythropoietin-overexpressed umbilical cord mesenchymal stem cells,apoptosis was significantly increased and H3K36me3 expression was significantly decreased.Transcriptome sequencing results showed that the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2 was reduced at mRNA transcription level(P<0.01).(8)It is concluded that erythropoietin-overexpressed umbilical cord mesenchymal stem cells activated the expression of REST and TET3 by altering the kurtosis of H3K4me2 and upregulated the modification level of H3K36me3,which in turn regulated the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2,and facilitated neuronal survival.
3.Qualitative analysis of volatile organic components in chemicals using headspace gas chromatography-mass spectrometry
Ruibo MENG ; Weifeng RONG ; Jiawen HU ; Jiaheng HE ; Jing YUAN ; Jingjing QIU ; Zuokan LIN ; Ziqun ZHANG ; Banghua WU
China Occupational Medicine 2023;50(3):325-329
Objective To establish a rapid qualitative analysis method for volatile organic components in chemicals. Methods Headspace gas chromatography-mass spectrometry was used to qualitatively determine 19 volatile organic components, including benzene, 1,2-dichloroethane, and n-hexane, in chemicals. Different sample amounts, heating temperatures, heating times, and sample volumes were analyzed to assess their effects on detection results and optimize sampling conditions. Results Based on the set chromatography, the optimal sampling process of this method was as follows: 5.0 g sample in a 20.0 mL headspace bottle, incubated at 40 ℃ for 30 minutes in a constant-temperature drying incubator, and a 1.00 mL headspace gas injection. The within-run and between-run relative standard deviations of all components ranged from 0.00% to 21.05% and 0.00% to 33.33%, respectively. The samples stored in sealed glass containers were stable at room temperature for at least 60 days. Conclusion This method offers simplicity, good reproducibility, and stability, making it suitable for rapid qualitative analysis of volatile organic components in chemicals.
4.Simultaneous determination of six aniline compounds in workplace air by high performance liquid chromatography
Ruibo MENG ; Jingjing QIU ; Weifeng RONG ; Weimin XIE ; Jing YUAN ; Jiaheng HE ; Jiawen HU ; Ziqun ZHANG ; Zuokan LIN ; Fan WU ; Banghua WU
China Occupational Medicine 2023;50(4):441-446
Objective To establish a high performance liquid chromatography (HPLC) method for simultaneous determination of six aniline compounds (ADs) in workplace air. Methods GDH-1 air sampling tube was used to collect six co-existing ADs such as aniline, o-toluidine, N-methylaniline, m-methylaniline, p-methylaniline and N,N-dimethylaniline in the vapor and aerosol of workplace air. The samples were desorbed and eluted using a methanol solution containing 1.00% ammonia water, followed by separation on a C18 chromatographic column and detection using a diode array detector. Results The quantification range of the method was 0.19 -253.50 mg/L, with the correlation coefficient of 0.999 9 for all six ADs. The minimum detection range was 0.02-0.06 mg/m3, and the minimum quantitation range was 0.04-0.19 mg/m3 [both calculated for a 15.0 L sample with a desorption (elution) solution volume of 3.00 mL]. The average desorption and elution efficiencies were 92.15%-104.41% (silica gel) and 94.29%-104.29% (filter membrane). The intra-assay relative standard deviation (RSD) ranged from 0.90%-9.72% (silica gel) and 0.57%-6.96% (filter membrane). The inter-assay RSD ranged from 2.03%-9.78% (silica gel) and 2.50%-8.62% (filter membrane). The samples were stable at room temperature for seven days. Conclusion This method can be used for the simultaneous determination of six ADs in workplace air.
5.Dl-3-N-butylphthalide alleviates renal ischemia-reperfusion injury by down-regulating NF-κB signaling pathway and inhibiting cell pyroptosis in rat models
Ruibo ZHANG ; Kaiwen SHEN ; Qiang WANG ; Qiang YUAN ; Jun SHEN
Organ Transplantation 2023;14(4):539-
Objective To elucidate the mechanism of dl-3-N-butylphthalide (NBP) on renal ischemia-reperfusion injury (IRI) in rat models. Methods Forty SD rats were randomly divided into the sham operation group (Sham group), model group (IRI group), NF-κB inhibitor pyrrolidine dithiocarbamate group (PDTC group), low-dose NBP group (NBP-L group) and high-dose NBP group (NBP-H group), with 8 rats in each group. Serum creatinine (Scr), serum cystatin C(Cys-C), blood urea nitrogen (BUN) and serum interleukin (IL)-1β and IL-18 levels were detected in all groups. Pathological injury of renal tissues in each group was observed by Hematoxylin-eosin (HE) staining. The expression levels of inflammatory factors and nuclear factor (NF)-κB signaling pathway and cell pyroptosis-related proteins in renal tissues were measured by Western blot and immunohistochemical staining. Results Compared with the Sham group, renal tissue injury was more severe, and the levels of Scr, Cys-C, BUN and serum IL-1β and IL-18 were all up-regulated in the IRI group. Western blot showed that the relative expression levels of NOD-like receptor protein (NLRP3), Gasdermin D(GSDMD), cysteinyl aspartate specific proteinase (Caspase)-1, IL-18, IL-1β, NF-κB p65 and p-NF-κB p65 proteins were all up-regulated, and immunohistochemical staining revealed that the expression levels of NF-κB p65 and p-NF-κB p65, IL-1β, IL-18 and NLRP3 proteins were all up-regulated in the IRI group. Compared with the IRI group, renal tissue injury was alleviated, and the levels of Scr, Cys-C, BUN and serum IL-18 and IL-1β were down-regulated in the PDTC, NBP-L and NBP-H groups. Western blot showed that the expression levels of NLRP3, GSDMD, Caspase-1, IL-1β, IL-18, NF-κB p65 and p-NF-κB p65 proteins were down-regulated, and immunohistochemical staining indicated that the expression levels of NF-κB p65, p-NF-κB p65, IL-1β, IL-18 and NLRP3 proteins were down-regulated in the PDTC, NBP-L and NBP-H groups, respectively. Compared with the NBP-L group, renal tissue injury was mitigated, and the levels of Scr, Cys-C, BUN, serum IL-18 and IL-1β were all down-regulated in the NBP-H group. Western blot showed the expression levels of NLRP3, GSDMD, Caspase-1, IL-1β, IL-18, NF-κB p65 and p-NF-κB p65 proteins were down-regulated in the NBP-H group. Immunohistochemical staining indicated that the expression levels of NF-κB p65, p-NF-κB p65, IL-1β, IL-18 and NLRP3 proteins were down-regulated in the NBP-H group. Conclusions NBP may down-regulate the activity of NF-κB/NLRP3 signaling pathway and reduce the expression levels of cell pyroptosis-related proteins and inflammatory factors after renal IRI, thereby suppressing cell pyroptosis and alleviating renal IRI.
6.Research progress of NF-κB signaling pathway in kidney ischemia-reperfusion injury
Ruibo ZHANG ; Kaiwen SHEN ; Qiang YUAN ; Qiang WANG ; Jun SHEN
Organ Transplantation 2022;13(3):349-
Kidney ischemia-reperfusion injury (IRI) is the major cause of poor prognosis after kidney transplantation and partial nephrectomy. Besides, it is also a critical pathophysiological process of acute kidney injury. Consequently, the prevention and treatment of kidney IRI are of significance to improve clinical prognosis of recipients undergoing kidney transplantation. However, the mechanism underlying IRI is complex, and the exact mechanism remains elusive. Inflammation, as one of the main pathogenesis of IRI, plays a significant role in IRI-induced kidney injury. Nuclear factor (NF)-κB, as a rapid response transcription factor, has been proven to be involved in the regulation of inflammation during kidney IRI. Therefore, in this article, the structure of NF-κB, the activation pattern of NF-κB signaling pathway, the regulatory mechanisms of NF-κB upstream and downstream signaling pathways in kidney IRI were reviewed, and the role of NF-κB signaling pathway in kidney IRI was investigated, aiming to provide novel clinical ideas for the prevention and treatment of kidney IRI.
7.Role of NLRP3 inflammasome and related inflammatory signaling pathways in renal ischemia-reperfusion injury
Qiang YUAN ; Kaiwen SHEN ; Ruibo ZHANG ; Jun SHEN
Organ Transplantation 2021;12(2):177-
Renal ischemia-reperfusion injury (IRI) commonly occurs in renal transplantation, which is an important pathophysiological process that causes acute renal failure and severely affects clinical prognosis of the recipients. Inflammatory response plays a critical role in the pathogenesis and pathological process of IRI. Activated NOD-like receptor protein 3(NLRP3) inflammasome can mediate the maturation and release of various pro-inflammatory cytokines, thereby regulating the inflammatory response and relevant cell functions. In this article, the mechanism underlying NLRP3 inflammasome and its related inflammatory signaling pathway in renal IRI were reviewed, aiming to provide novel ideas for clinical prevention and treatment of renal IRI.
8.Artesunate alleviates renal ischemia-reperfusion injury in rats by inhibiting pyroptosis via NLRP3 inflammasome
Qiang YUAN ; Kaiwen SHEN ; Ruibo ZHANG ; Qiang WANG ; Yuting WU ; Jun SHEN
Organ Transplantation 2021;12(6):733-
Objective To investigate the effect and mechanism of artesunate on renal ischemia-reperfusion injury (IRI) in rat. Methods Twenty-five SD rats were randomly divided into the sham operation group (Sham group), model group (IRI group), low-dose artesunate group (ART-L group), high-dose artesunate group (ART-H group) and NLRP3 inflammasome inhibitor group (INF39 group), with 5 rats in each group. The levels of serum creatinine (Scr), blood urea nitrogen (BUN) and pathological damage of renal tissue were analyzed. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum were quantitatively measured. The expression level of IL-1β in renal tissues was determined by immunohistochemical staining. The expression levels of pyroptosis-related proteins were detected by fluorescent staining and Western blot. Results Compared with the Sham group, the levels of Scr and BUN were higher, the renal tissue injury was aggravated, the expression levels of TNF-α, IL-6 and IL-1βwere higher, and the expression levels of kidney injury molecule (KIM-1), pyroptosis-related protein NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase (Caspase-1), Gasdermin D (GSDMD) and IL-1β proteins were higher in the IRI group. Compared with the IRI group, the levels of Scr and BUN were decreased, the renal tissue injury was mitigated, the expression levels of TNF-α, IL-6 and IL-1β were down-regulated, and the expression levels of KIM-1, NLRP3, Caspase-1, GSDMD and IL-1β proteins were down-regulated in the ART-L, ART-H and INF39 groups. Conclusions Artesunate may inhibit pyroptosis induced by NLRP3 inflammasome, down-regulate the expression levels of pyroptosis -related proteins, reduce the release of inflammatory factors after renal IRI and alleviate renal IRI.
9.The progress on the mechanism of NLRP3 inflammasome in renal ischemia-reperfusion injury
Qiang YUAN ; Kaiwen SHEN ; Ruibo ZHANG ; Jun SHEN
Chinese Journal of Urology 2021;42(7):554-558
Renal ischemia-reperfusion injury often occurs during operations that need to block the blood supply of the kidneys. It is an important pathophysiological process that affects the recovery of kidney function and causes acute renal failure. Activated NLRP3 inflammasome can mediate the maturation and release of a variety of pro-inflammatory factors, and participate in the inflammatory response in renal ischemia-reperfusion injury, thereby regulating body inflammation and related cell functions. This article summarized how NLRP3 inflammasome mediated the occurrence of inflammation in renal ischemia reperfusion injury, and further provied a new basis for the prevention and treatment of renal ischemia-reperfusion injury.
10.Compilation and reliability and validity test of hospital nursing treatment capacity scale for patients with nuclear radiation damage
Ruibo LI ; Rufu JIA ; Qi YE ; Chenghong ZHU ; Xiuyan SUN ; Wei ZHANG
Chinese Journal of Radiological Health 2021;30(5):611-615
Objective The nursing treatment ability scale of patients with nuclear radiation damagein the hospital was developed to provide an evaluation basis for improving the nursing ability of nurses with nuclear radiation damage. Methods The scale was prepared by literature review, expert interview and expert consultation, and a total of 330 clinical nurses from a third-class hospital was randomly selected as the research objects. The scales were issued for item analysis and reliability and validity test. Results The scales were divided into 6 dimensions, including basic knowledge of nuclear radiation damage, specialized equipment use ability, specialized ward management ability, basic nursing ability, specialized nursing ability and self-ability recognition, with 51 items. After exploratory factor analysis, there were 6 principal components, and the cumulative interpreted variance was 70.757%. The χ2, df, χ2/df, CFI, IFI, TLI, NFI, PNFI, PCFI, RMSEA fitting indexes of confirmatory factor analysis were all acceptable. Cronbach's α coefficient was 0.976, the retest reliability was 0.823, and the S-CVI (S-CVI/UA) was 0.84. The evaluation content validityS-CVI (S-CVI/AVE) was 0.98, and the content validity I-CVI of the item level was 0.78~1.00. Conclusion The items and dimension Settings of this scale have been tested, and all indicators met the requirements. The reliability and validity test results were good. It can be used as a scale for preliminary evaluation of hospital nursing ability of patients with nuclear radiation damage.

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