Objective To investigate the annual effective doses to radiation workers caused by radon concentrations in the underground workplaces of medical institutions, and to provide a scientific basis for the prevention and control of indoor radon in underground places. Methods A typical sampling method was used to select 5-30 medical institutions in each of Hunan, Jiangxi, Guizhou, Hubei, and Sichuan provinces. A total of 66 monitoring points in 66 medical institutions were selected. The indoor radon concentrations in underground workplaces were measured cumulatively using CR-39 solid nuclear track detectors. The radiation dose to radiation workers was estimated according to the method outlined in the Requirements for control of indoor radon and its progeny (GB/T 16146—2015). The Kruskal-Wallis H test was used to compare the differences in indoor radon concentrations between different provinces. Results The average indoor radon concentration in the underground workplaces of 66 medical institutions was 69.8 Bq/m³, with the highest being 147.6 Bq/m³. The average indoor radon concentrations in the underground workplaces of medical institutions in Sichuan, Guizhou, Hubei, Jiangxi, and Hunan were 72.1, 83.2, 66.6, 88.4, and 61.5 Bq/m³, respectively. The annual effective doses to radiation workers caused by radon concentrations in underground workplaces were 0.57-0.83 mSv, with an average of 0.69 mSv. There was a significant difference in radon concentrations among provinces (P < 0.05). Conclusion The indoor radon concentrations and personnel exposure doses in the underground workplaces of monitored medical institutions comply with national control standards. However, continuous monitoring and necessary indoor radon prevention and control measures are still needed.

OBJECTIVES: Pyroptosis plays a critical role in tubulointerstitial lesions of diabetic kidney disease (DKD). Annexin A2 (ANXA2) is involved in cell proliferation, apoptosis, and adhesion and may be closely related to DKD, but its specific mechanism remains unclear. This study aims to investigate the role and molecular mechanism of ANXA2 in high glucose-induced pyroptosis of renal tubular epithelial cells, providing new targets for DKD prevention and treatment. METHODS: Human renal tubular epithelial HK-2 cells were divided into a normal glucose group (5.5 mmol/L), a high glucose group (30.0 mmol/L), and a osmotic control group (24.5 mmol/L mannitol+5.5 mmol/L glucose). ANXA2 expression was modulated by overexpression of plasmids and small interfering RNA (siRNA). Cell proliferation was measured by 5-ethynyl-2'-deoxyuridine (EdU) assay, apoptosis by flow cytometry, and ANXA2, p50, and p65 subcellular localization by immunofluorescence. Western blotting was employed to detect α-smooth muscle actin (α-SMA), fibronectin (FN), and collagen type IV (Col-IV). Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting were used to analyze nuclear factor-κB (NF-κB) subunits p50/p65 and the pyroptosis pathway factors NLR family Pyrin domain containing 3 (NLRP3), caspase-1, inferleukin (IL)-1β, and IL-18. Protein interactions between ANXA2 and p50/p65 were examined by co-immunoprecipitation, while chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were used to examine NF-κB binding to the ANXA2 promoter. RESULTS: High glucose upregulated ANXA2 expression and promoted its nuclear translocation (P<0.01). High glucose reduced cell proliferation, increased apoptosis, and elevated α-SMA, FN, and Col-IV expression (all P<0.05); ANXA2 overexpression aggravated these effects (all P<0.05), while ANXA2 knockdown reversed them (all P<0.05). High glucose activated NF-κB and increased NLRP3, caspase-1, L-1β, and IL-18 mRNA and protein expression (all P<0.05); ANXA2 overexpression further enhanced this, whereas knockdown suppressed NF-κB activation and downstream factors (all P<0.05). Co-immunoprecipitation confirmed ANXA2 directly binds the NF-κB subunit p65. ChIP assays revealed p65 binds specifically to ANXA2 promoter regions (ChIP-2, ChIP-4, and ChIP-6), and luciferase activity in corresponding mutant constructs (M2, M4, and M6) was significantly increased versus controls (all P<0.05), confirming positive transcriptional regulation of ANXA2 by p65. CONCLUSIONS ANXA2 and NF-κB form a positive feedback loop that sustains NLRP3 inflammasome activation, promotes pyroptosis pathway activation, and aggravates high glucose-induced renal tubular epithelial cell injury. Targeting ANXA2 or blocking its interaction with p65 may be a novel strategy to slow DKD progression.
Humans ; Pyroptosis/drug effects* ; Annexin A2/physiology* ; Epithelial Cells/cytology* ; Kidney Tubules/cytology* ; Glucose/pharmacology* ; Diabetic Nephropathies/metabolism* ; NF-kappa B/metabolism* ; Cell Line ; Cell Proliferation ; Transcription Factor RelA/metabolism* ; Feedback, Physiological

Ischemic stroke (IS) is a globally life-threatening disease. Presently, few therapeutic medicines are available for treating IS, and rt-PA is the only drug approved by the US Food and Drug Administration (FDA) in the US. In fact, many agents showing excellent neuroprotection but no blood flow-improving activity in animals have not achieved ideal clinical efficacy, while thrombolytic drugs only improving blood flow without neuroprotection have limited their wider application. To address these challenges and meet the huge unmet clinical need, we have designed and identified a novel compound AAPB with dual effects of neuroprotection and cerebral blood flow improvement. AAPB significantly reduced cerebral infarction and neural function deficit in tMCAO rats, pMCAO rats, and IS rhesus monkeys, as well as displayed exceptional safety profiles and excellent pharmacokinetic properties in rats and dogs. AAPB has now entered phase I of clinical trials fighting IS in China.

Cuproptosis, a recently identified form of regulated cell death triggered by excess intracellular copper, has emerged as a promising cytotoxic strategy for cancer therapy. However, the therapeutic efficacy of copper ionophores such as elesclomol (ES) is often hindered by cellular copper homeostasis mechanisms that limit copper influx and cuproptosis induction. To address this challenge, we developed a nanoagent utilizing outer membrane vesicle (OMV) derived from Akkermansia muciniphila (Akk) for co-delivery of antioxidant 1 copper chaperone (Atox1)-targeting siRNA and ES (siAtox1/ES@OMV) to tumors. In vitro, we demonstrated that Atox1 knockdown via siRNA significantly disrupted copper export mechanisms, resulting in elevated intracellular copper levels. Simultaneously, ES facilitated efficient copper influx and mitochondrial transport, leading to Fe-S cluster depletion, increased proteotoxic stress, and robust cuproptosis. In vivo, siAtox1/ES@OMV achieved targeted tumor delivery and induced pronounced cuproptosis. Furthermore, leveraging the immunomodulatory properties of OMVs, siAtox1/ES@OMV promoted T-cell infiltration and the activation of tumor-reactive cytotoxic T cells, enhancing tumor immune responses. The combination of siAtox1/ES-induced cuproptosis and immunogenic cell death synergistically suppressed tumor growth in both subcutaneous breast cancer and orthotopic rectal cancer mouse models. This study highlights the potential of integrating copper homeostasis disruption with a copper ionophore using an immunomodulatory OMV-based vector, offering a promising combinatorial strategy for cancer therapy.

OBJECTIVE To explore the protective effect and mechanism of gallic acid(GA)on paclitaxel-induced peripheral neuropathy.METHODS The primary dorsal root ganglion(DRG)was extracted to establish an in vitro model of paclitaxel-induced DRG cell injury;CCK-8 assay was used to evaluate the neuroprotective effects of GA on this model.The paclitaxel-induced neuro-pathic pain model in C57BL/6J mice was established,and the behavioral test was used to analyze effects of GA alleviating paclitaxel-induced peripheral neuropathy and to explore whether GA alleviated paclitaxel-induced peripheral neuropathic pain by inhibiting the activation of NLRP3 inflammatory and reducing the release of pro-inflammatory cytokine.qPCR was used to detect the expression lev-els of NLRP3,Caspase-1,and IL-1β mRNA in DRG cells;Western blot was used to detect the change of protein levels of NLRP3,Caspase-1,and IL-1β;and the fluorescence intensity of NLRP3,Caspase-1,and IL-1β proteins was analyzed by immunofluores-cence staining.RESULTS GA exerts a protective effect and improved the survival rate of paclitaxel-stimulated DRG cells;reduced mechanical,cold and thermal pain hyperalgesia in mice;effectively reversed the elevated expression levels of NLRP3 and Caspase-1 in paclitaxel-stimulated DRG cells and reduced the release of pro-inflammatory cytokine IL-1β,which ameliorated the neuroinflam-matory response induced by paclitaxel.CONCLUSION Gallic acid improves paclitaxel-induced neuroinflammation by regulating the NLRP3 inflammasome pathway.The NLRP3/Caspase-1/IL-1β pathway is one of the potential mechanisms for the treatment of pacli-taxel-induced peripheral neuropathy.

Objective:To understand the current status of gastroscopy in diagnosing gastric lesions in general population, and to recommend the optimal age for the first gastroscopy and intervals for repeated gastroscopy.Methods:The gastroscopy records of residents aged 18-80 years in Yinzhou District of Ningbo, Zhejiang Province, between April 2010 and December 2021 were analyzed retrospectively. The detections of gastric lesions across different years, age and genders were described. Goodness of fit tests were applied to compare the differences in detection rates of different lesions in first-time endoscopy in different age groups and different populations. Generalized additive models were used to fit the trend of age specific gastric lesion detection rate explore the optimal age for gastroscopy. The appropriate gastroscopy intervals were determined according to the progress of the gastric lesions detected in repeated gastroscopy.Results:A total of 237 751 participants with 344 398 gastroscopy records were included in analyses. A total of 5 597 cases of chronic atrophic gastritis (CAG), 9 796 cases of intestinal metaplasia (IM), 165 cases of low-grade intraepithelial neoplasia (LGIN), 52 cases of high-grade intraepithelial neoplasia (HGIN) and 435 cases of gastric cancer were detected by the first gastroscopy. The overall detection rate of gastric lesions increased significantly in age group 45-70 years, and remained stable after 70 years old, with LGIN and HGIN showing notable increases at 50 and 55 years old, respectively. Repeated gastroscopy detected CAG, IM, LGIN, and HGIN at a higher rate compared with the first gastroscopy. Normal/superficial gastritis progressed in 3-5 years, whereas CAG or more severe lesions progressed in 1-6 years.Conclusion:Gastroscopy is recommended for general population aged 45 years and above. Furthermore, gastroscopy can be performed every 3-5 years for individuals with normal endoscopy results and once a year for patients with CAG or more severe gastric lesions.

To explore the value of CNVPLUS ?-array in the diagnosis of the DMD gene. A retrospective study was performed on 96 children who were clinically diagnosed with Duchenne or Becker muscular dystrophies(DMD/BMD) at the Department of Pediatric Endocrinology and Genetics of Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine from January 2014 to March 2023. DNA was extracted from these children′s peripheral blood and divided into two parts. Variations of the DMD gene were detected by using CNVPLUS ?-array and sequential testing of MLPA—NGS—Sanger. In the sequential method, single exon deletions detected by MLPA were first verified by polymerase chain reaction (PCR) and then were tested by Sanger′s sequencing if PCR results were normal. The results showed that, among 96 samples, 91 cases with the pathogenic variation of the DMD gene were detected by the CNVPLUS ?-array, including 76 cases with large deletion/duplication (copy number variation, CNV) and 15 cases with small variation (single nucleotide variant or small insertion/deletion, SNV/Indel). All samples were tested and diagnosed within 5 days. In contrast, 76 cases with pathogenic CNV and 20 cases with pathogenic SNV/Indel were detected in the DMD gene by sequential method. However, all of the experiments and diagnoses were completed within 48 days. Moreover, 5 cases with SNV/Indel in the DMD gene were correctly clustered after the operation mode was optimized. In summary, as a new micro-array integrating CNV and SNV probes, CNVPLUS ?-array can detect CNV and SNV/Indel in the DMD gene simultaneously while the application of CNVPLUS ?-array could save a lot of time and manpower. CNVPLUS ?-array had an excellent diagnostic performance for CNV of the DMD gene. As for SNV/Indel, the diagnostic performance was slightly poor and the operation mode should be optimized. If necessary, other testing technologies should be supplemented to reduce the risk of missed diagnosis.

Aim To explore the effect of Pueraria Lobata Flowers Extract(PFE)on lipid accumulation in mac-rophage-derived foam cells.Methods The concentration of PFE in THP-1-derived foam cells was screened by MTT,intracellular lipid accumulation was detected by oil red O staining and cholesterol detection kit,intracellular cholesterol ef-flux levels were detected by cholesterol efflux assay kit,RT-qPCR and Western blot were used to analyze mRNA and pro-tein expression.Results PFE significantly reduced lipid accumulation in THP-1-derived foam cells.PFE did not affect the mRNA expression of CD36,scavenger receptor-A Ⅰ(SR-A Ⅰ),sterol regulatory element-binding protein 2(SREBP2),3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR),but it could upregulate the mRNA and protein expres-sion levels of ATP-binding cassette transporter A1(ABCA1)(P＜0.05),and promote the intracellular cholesterol efflux of macrophage-derived foam cells(P＜0.01).PFE could activate the activity of peroxisome proliferator-activated receptor y(PPARγ)(P＜0.01)and upregulate the mRNA and protein expression levels of PPARγ(P＜0.05).Compared with the PFE control group,the expression of PPARγ and ABCA1 proteins decreased and cholesterol efflux decreased after GW9662 treatment(all P＜0.01).Conclusion PFE could significantly prevent the lipid accumulation in THP-1-derived foam cells and inhibit the formation of foam cells by upregulating ABCA1 expression and cholesterol efflux mediated by PPARγ.

To explore the value of CNVPLUS ?-array in the diagnosis of the DMD gene. A retrospective study was performed on 96 children who were clinically diagnosed with Duchenne or Becker muscular dystrophies(DMD/BMD) at the Department of Pediatric Endocrinology and Genetics of Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine from January 2014 to March 2023. DNA was extracted from these children′s peripheral blood and divided into two parts. Variations of the DMD gene were detected by using CNVPLUS ?-array and sequential testing of MLPA—NGS—Sanger. In the sequential method, single exon deletions detected by MLPA were first verified by polymerase chain reaction (PCR) and then were tested by Sanger′s sequencing if PCR results were normal. The results showed that, among 96 samples, 91 cases with the pathogenic variation of the DMD gene were detected by the CNVPLUS ?-array, including 76 cases with large deletion/duplication (copy number variation, CNV) and 15 cases with small variation (single nucleotide variant or small insertion/deletion, SNV/Indel). All samples were tested and diagnosed within 5 days. In contrast, 76 cases with pathogenic CNV and 20 cases with pathogenic SNV/Indel were detected in the DMD gene by sequential method. However, all of the experiments and diagnoses were completed within 48 days. Moreover, 5 cases with SNV/Indel in the DMD gene were correctly clustered after the operation mode was optimized. In summary, as a new micro-array integrating CNV and SNV probes, CNVPLUS ?-array can detect CNV and SNV/Indel in the DMD gene simultaneously while the application of CNVPLUS ?-array could save a lot of time and manpower. CNVPLUS ?-array had an excellent diagnostic performance for CNV of the DMD gene. As for SNV/Indel, the diagnostic performance was slightly poor and the operation mode should be optimized. If necessary, other testing technologies should be supplemented to reduce the risk of missed diagnosis.

Objective:To discuss the effect of adipose-derived stem cell-derived exosomes(ADSC-Exos)on the migration ability of the macrophages RAW264.7,and to clarify its role in promoting function of the macrophages.Methods:The adipose tissue adjacent to epididymis of the SD rats was isolated to perform primary culture of the adipose-derived stem cells(ADSCs).The adipogenic and osteogenic differentiation induction was conducted,and the multidirectional differentiation potential of the ADSCs was detected by oil Red O and Alizarin red staining.Western blotting and immunofluorescence methods were used to detect the positive expressions of the ADSCs markers CD29 and CD44;the ADSC-Exos were extracted by Exos isolation kit,and the morphology,size,and distribution of particle size of the ADSC-Exos were examined by transmission electron microscope and nanoparticle tracking analyzer;the expression levels of exosome-specific markers CD9 and TSG101 proteins in the ADSC-Exos were detected by Western blotting method;the uptake of ADSC-Exos by the macrophages was observed by tracing method.The macrophages RAW264.7 were divided into control group,10 mg·L-1 ADSC-Exos group,20 mg·L-1 ADSC-Exos group,and 40 mg·L-1 ADSC-Exos group.The activities of the macrophages in various groups were detected by 5-ethynyl-2'-deoxyuridine(EdU)staining;the number of migration macrophages in various groups was detected by Transwell chamber assay;the adhesion of macrophages in various groups was observed by fluorescence microscope.Results:After 24 h of primary culture,the ADSCs adhered to the wall and exhibited scattered,elongated shapes;after 7 d of culture,the adherent cells showed a comb-like,vortex-like orderly arrangement,resembling fibroblasts;after 10 passages,the irregular morphology of the ADSCs and decreased proliferation rate were found.The isolated ADSCs showed potential for the osteogenic and adipogenic differentiation,and the expressions of CD29 and CD44 proteins were positive.The transmission electron microscope observation resuls showed that the ADSC-Exos appeared disc-shaped,and the main peak of particle size distribution was around 132 nm.The CD9 and TSG101 proteins were positively expressed in the ADSC-Exos,indicating successful extraction.The fluorescence microscope results showed red fluorescence signals around the nuclei of the RAW264.7 cells,indicating the uptake of ADSC-Exos by the macrophages.Compared with control group,the rates of EdU positive cells in 10,20,and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05);compared with 10 mg·L-1 ADSC-Exos group,the rate of EdU positive cells in 20 mg·L-1 ADSC-Exos group was significantly increased(P<0.05).Compared with control group,the numbers of migration cells in 10,20,and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05);compared with 10 mg·L-1 ADSC-Exos group,the numbers of migration cells in 20 and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05).Compared with control group,the numbers of the adherent macrophages in 10,20,and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05);compared with 10 mg·L-1 ADSC-Exos group,the number of adherent macrophages in 20 mg·L-1 ADSC-Exos group was significantly increased(P<0.05).Conclusion:The ADSC-Exos can be internalized by the macrophages and they can enhance the migration ability of the macrophages by affecting the cell adhesion.