1.Construction of PD-L1hitol-DC derived from bone marrow of DA rats and identification of its immunological function
Zhiqi YANG ; Peibo HOU ; Lang WU ; Jing LIU ; Yang DING ; Minghao LI
Organ Transplantation 2025;16(1):83-90
Objective To construct programmed cell death protein-ligand 1(PD-LI)hi tolerogenic dendritic cell (tol-DC) derived from bone marrow of DA rats and identify its immunological function. Methods DA rat bone marrow cells were extracted, combined with recombinant mouse granulocyte macrophage colony-stimulating factor and recombinant mouse interleukin (IL)-4, and cultured for 6 days in vitro to induce the differentiation of bone marrow cells into immature dendritic cells (imDC). Lipopolysaccharide was used to stimulate cell maturation and cultured for 2 days to collect mature dendritic cells (mDC). PD-L1 lentiviral vector virus stock solution or equivalent dose lentiviral stock solution was added, and PD-L1hitol-DC and Lv-imDC were collected after culture for 2 days. The morphology of PD-L1hitol-DC was observed by inverted phase contrast microscope and transmission electron microscope. Real-time fluorescence quantitative reverse transcription polymerase chain reaction, Western blotting and flow cytometry were used to detect the expression level of specific markers on cell surface. CD8+T cells derived from Lewis rat spleen were co-cultured with imDC, mDC, Lv-imDC and PD-L1hitol-DC, respectively. The levels of inflammatory factors in the supernatant of each group were detected by enzyme-linked immunosorbent assay. The apoptosis of T cells and the differentiation of regulatory T cells (Treg) in each group were analyzed by flow cytometry. Results The morphology of PD-L1hitol-DC modified by PD-L1 gene was consistent with tol-DC characteristics, and the expression levels of CD80, CD86 and major histocompatibility complex (MHC) on the surface were low. After mixed culture with CD8+ T cells, the levels of IL-10 and transforming growth factor (TGF) -β1 in the supernatant of PD-L1hitol-DC group were higher, the levels of tumor necrosis factor (TNF) -α and IL-17A were lower, and the apoptosis of T cells and Treg differentiation were increased. Conclusions Overexpression of PD-L1 through lentiviral vectors may successfully induce the construction of bone-marrow derived PD-L1hitol-DC in DA rats, promote the secretion of anti-inflammatory factors and T cell apoptosis, induce the differentiation of Treg, and inhibit the immune response of allogeneic CD8+T cells, which provides experimental basis for the next organ transplantation immune tolerance study.
2.Protective effect of Shenfu injection against neonatal hypoxic-ischemic brain injury by inhibiting the ferroptosis
Xiaotong Zhang ; Meng Zhang ; Gang Li ; Yang Hu ; Yajing Xun ; Hui Ding ; Donglin Shen ; Ming Wu
Acta Universitatis Medicinalis Anhui 2025;60(1):31-40
Objective :
To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis.
Methods :
An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression.
Results :
Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01).
Conclusion
Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.
3.Research progress on pathogenesis and exercise intervention of adolescent with upper cross syndrome
YIN Ding, WANG Hui, ZHAI Suo, CHEN Miao
Chinese Journal of School Health 2025;46(1):144-147
Abstract
Upper cross syndrome (UCS) is an abnormal body posture phenomenon. The detection rate of UCS in adolescents is high in China, which has become a serious public health problem threatening the health of adolescents. Exercise therapy has a positive effect on the prevention and treatment of UCS.The article reviews the pathological mechanisms of muscle imbalance, muscle interaction inhibition, and neural control related to the occurrence and development of UCS both domestically and internationally in recent years.And it elucidates the effects and corresponding mechanisms of exercise interventions such as gentle exercise, stretching exercise and physical function training on UCS, so as to provide help for the correction of UCS.
4.HAN Mingxiang's Experience in Staged and Syndrome-Based Treatment of Chronic Obstructive Pulmonary Disease
Jian DING ; Hui TAO ; Gang CHENG ; Weizhen GUO ; Zegeng LI ; Ya MAO ;
Journal of Traditional Chinese Medicine 2025;66(8):780-785
This paper summarizes Professor HAN Mingxiang's clinical experience in treating chronic obstructive pulmonary disease (COPD). He believes that the key pathomechanism of COPD in the acute exacerbation stage is the invasion of external pathogens triggering latent illness, while lung qi deficiency is the primary mechanism in the stable stage. The core pathological factors throughout disease progression are deficiency, phlegm, and blood stasis. Treatment emphasizes a staged and syndrome-based approach. During the acute exacerbation stage, for wind-cold invading the lung syndrome, the self-formulated Sanzi Wenfei Decoction (三子温肺汤) is used to relieve the exterior, dispel cold, warm the lung, and resolve phlegm. For phlegm-dampness obstructing the lung syndrome, Huatan Jiangqi Fomulation (化痰降气方) is prescribed to warm the lung, transform phlegm, descend qi, and calm wheezing. For phlegm-heat obstructing the lung syndrome, Qingfei Huatan Fomulation (清肺化痰方) is applied to clear heat, resolve phlegm, moisten the lung, and stop coughing. For phlegm and blood stasis interlocking syndrome, Qibai Pingfei Fomulation (芪白平肺方) is used to tonify qi, resolve phlegm, and activate blood circulation to remove stasis. During the stable stage, for lung qi deficiency syndrome, Shenqi Wenfei Decoction (参芪温肺汤) is employed to warm the lung, tonify qi, resolve phlegm, and eliminate turbidity. For lung-spleen qi deficiency syndrome, Shenqi Buzhong Decoction (参芪补中汤) is utilized to strengthen the spleen, tonify qi, and reinforce metal (lung) from earth (spleen). For lung-kidney deficiency syndrome, Shenqi Tiaoshen Fomulation (参芪调肾方) is prescribed to tonify the lung, warm yang, and regulate kidney function to calm wheezing. These strategies provide insights into the traditional Chinese medicine treatment of COPD.
8.Exposure to indoor chemical pollutants and associated health risk assessment of residents in Shijiazhuang city
Siwei ZHANG ; Yishan DING ; Xixi DU ; Fengge CHEN ; Junwang TONG ; Hui MA
Journal of Environmental and Occupational Medicine 2025;42(6):706-711
Background Indoor air quality directly affects people's health, especially the impact of chemical pollutants in residential indoor air on children and the elderly is more significant. Objective To understand the pollution status of common chemical pollutants in residential indoor air in Shijiazhuang, evaluate the health risks of chemical pollutants to school-age children and the elderly, and provide reference for controlling indoor pollution in residential environment. Methods Using stratified random sampling, a total of 60 households were selected from 2 urban areas and 1 surrounding rural area in Shijiazhuang City, specifically in July 2023 (non-heating season) and December 2023 (heating season), respectively. Relevant data was collected through on-site sampling [including CO, CO2, PM10, PM2.5, NO2, SO2, O3, ammonia, formaldehyde, benzene, toluene, xylene, total volatile organic compounds (TVOC), trichloroethylene, and tetrachloroethylene] and survey questionnaires. The pollutant concentrations were evaluated following the Standards for indoor air quality of GB/T 18883-2022, and the inhalation exposure risks of the target population were assessed based on the health risk assessment method. Results In the indoor air of the urban and rural residence in Shijiazhuang City, except for CO, NO2, SO2, toluene, and xylene, which did not exceed the standard limits, other pollutants showed varying degrees of exceedance. The non-qualified rates of PM10, PM2.5, and CO2 in the urban areas were higher than those in the rural areas (P < 0.05). The seasonal difference analysis showed that the non-qualified rates of PM2.5, PM10, CO2, trichloroethylene, and tetrachloroethylene in the urban areas were higher in the heating season than in the non-heating season (P<0.05); the non-qualified rates of ammonia and formaldehyde in the rural areas increased significantly in the non-heating season(P<0.05). The health risk assessment indicated that the maximum hazard quotient (HQ) of tetrachloroethylene for the elderly exceeded 1, while the HQ values for ammonia, formaldehyde, benzene, toluene, xylene, and trichloroethylene remained below 1 for either children or the elderly. For carcinogenic risks, the median carcinogenic risk (CR) of formaldehyde for school-aged children or the elderly fell within the range of 10−6-10−4, whereas the median CR values for benzene, trichloroethylene, and tetrachloroethylene were all below 10−6. Conclusion The primary indoor air pollutants exceeding the national standard limits in residential areas of Shijiazhuang City include CO2, PM10, TVOC, PM2.5, formaldehyde, ammonia, trichloroethylene, and tetrachloroethylene. The levels of these pollutants exhibit significant urban-rural and seasonal variations. Special attention should be paid to the non-carcinogenic risk of tetrachloroethylene to the elderly and the carcinogenic risks of formaldehyde to school-age children and the elderly.
9.Assoication between 24 hour activity time allocation and active health levels among college students in Yinchuan City
CHEN Miao, ZHAI Suo, DING Wenqing, YIN Ding
Chinese Journal of School Health 2025;46(7):950-955
Objective:
To explore the potential classification of 24 hour activity time allocation among college students in Yinchuan and its association with active health levels, so as to provide references for optimizing activity time allocation to enhance active health levels.
Methods:
From November 18 to December 6, 2024, a stratified cluster random sampling method was used to select 2 422 first and second year college students from full time undergraduate institutions in Yinchuan. The Chinese College Students 24 hour Movement Behaviors Questionnaire (24 h MBQ) and Active Health Behavior Scale were used to assess 24 hour activity time allocation and evaluate active health levels. Latent profile analysis (LPA) was employed to categorize activity types, and a binary Logistic regression analysis was conducted to analyze the relationship between active health levels and activity types.
Results:
A total of 1 087 students (44.9%) were found of meeting active health standards, and significant statistical differences were found in active health levels across different genders, grades, academic qualities, sources of origin and academic categories ( χ 2= 22.03 , 7.65, 25.50, 10.12, 43.44, all P <0.01). Moreover, significant statistical differences could also be found among college students 24 hour activity time across different genders, ages, grades, sources of origin, academic qualities, and academic categories ( t/Z/H/F=-5.70-111.39, P <0.05).The 24 hour activity time allocation was classified into four types:academic high ( 6.9 %), low activity rest (8.8%), light activity (67.8%), and high activity dynamic (16.4%). Significant statistical differences were observed in activity time allocation categories across different ages, academic qualities and academic categories ( χ 2=15.52-108.46, all P <0.05). Using the high activity dynamic type as a reference, the light activity type ( OR=0.39, 95%CI =0.31-0.50), low activity rest type ( OR=0.10, 95%CI =0.06-0.15), and academic high type ( OR=0.03, 95%CI =0.02-0.07) had lower active health levels among college students (all P <0.01).
Conclusion
There is a significant difference in 24 hour activity time allocation among college students in Yinchuan, and different activity types are associated with active health levels.
10.Subchronic exposure to benzoapyrene results in lung tissue cell damage caused by ferroptosis in mice
Chaoli ZHOU ; Shihan DING ; Hui HE ; Zhirui MA ; Jie CHEN ; Xingdi GUO ; Yi LYU ; Jinping ZHENG
Journal of Environmental and Occupational Medicine 2025;42(8):971-977
Background Exposure to benzo[a]pyrene (BaP) may impair lung function through various mechanisms; however, it remains uncertain whether BaP induces ferroptosis in lung tissue cells, resulting in lung function impairment. Objective To investigate the ferroptosis of lung tissue cells triggered by subchronic BaP exposure in mice and its correlation with lung injury, and to explore the function of ferroptosis in BaP-induced lung tissue damage. Method Seventy-two healthy 3-weeks-old male C57BL/6J mice were acclimatized for 1 week and then randomly divided into six groups: control group (corn oil 10 mL·kg−1), low-dose BaP group (2.5 mg·kg−1), medium-dose BaP group (5 mg·kg−1), high-dose BaP group (10 mg·kg−1), BaP+ferrostatin-1 (Fer-1) group (10 mg·kg−1+1 mg·kg−1), and Fer-1 group (1 mg·kg−1), with 12 mice each group. Corn oil and BaP were administered via gavage every other day, followed by an intraperitoneal injection of Fer-1 the subsequent day, throughout a period of 90 d. Whole-body plethysmography was applied to detect lung function; hematoxylin-eosin staining (HE) and Masson staining were used to observe lung tissue injury and fibrosis; microscopy of alveolar epithelial cells was conducted to reveal mitochondrial morphology; biochemical assays were used to measure the content of tissue iron, malondialdehyde (MDA), and glutathione (GSH), as well as the activity of glutathione peroxidase (GSH-Px); Western blotting and real-time quantitative PCR (RT-qPCR) analyses were performed to reveal the protein and mRNA expression of ferroptosis markers. Results Compared to the control group, the high-dose BaP group showed a significant increase in expiration time (Te) (P<0.01), and a significant decrease in ratio rate of achieving peak expiratory flow (Rpef), tidal volume (TVb), peak inspiratory flow (PIF), minute volume (MVb), and peak expiratory flow (PEF) (P<0.05 or 0.01). Based on the results of HE and Masson staining, partial destruction of alveolar structures, thickening of alveolar walls, infiltration of inflammatory cells, significant thickening of tracheal walls and a large deposition of collagen fibers in lung tissue were observed in the medium- and high-dose BaP groups. By microscopy, the alveolar epithelial cells exposed to low-dose BaP showed condensed chromatin, and the mitochondria exposed to medium and high-dose BaP showed wrinkles, increased mitochondrial membrane density, and diminished mitochondrial cristae. Compared to the control group, in the medium- and high-dose BaP groups, the lung tissue iron content and the expression levels of ACSL4 protein and mRNA significantly elevated (P<0.01 or 0.05), while the mRNA expression level of SLC7A11 significantly decreased (P<0.05); in the high-dose BaP group, the MDA content, COX2 protein, and PTGS2 mRNA expression levels significantly increased (P<0.05 or 0.01), GSH content and GSH-Px activity, GPX4 protein and mRNA expression levels, and the expression level of SLC7A11 protein significantly decreased (P<0.01 or 0.05). The ferroptosis inhibitor Fer-1 markedly reversed respiratory function, morphology, mitochondrial alterations, and the aforementioned ferroptosis-related biochemical indicators. Conclusion Subchronic exposure to BaP can induce ferroptosis in mice lung tissue cells, resulting in compromised lung function.


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