1.Advantages of modified ligation method for spinal cord injury modeling
Daohui LI ; Xiaoshuang XU ; Zhengtao LI ; Xinpeng TIAN ; Hangchuan BI ; Yuan LIU ; Yongwen DAI ; Lingqiang CHEN
Chinese Journal of Tissue Engineering Research 2025;29(2):379-384
BACKGROUND:Currently,different methods of model establishment have been derived from different injury modes of spinal cord injury.Traditional physical injury modeling methods have their own advantages and disadvantages,and there is a lack of more effective and stable animal models of spinal cord injury. OBJECTIVE:To establish a reproducible,controllable,trauma-free,low-mortality,more stable,widely applicable,and short-term postoperative care rat model of spinal cord injury. METHODS:Forty Sprague-Dawley rats with similar body mass and ages were randomly divided into a control group and an improved group,with 20 rats in each group.Animal models of spinal cord injury in the control group were constructed using a clip model method,while the improved group used a modified ligation method based on the compression method to make the spinal cord injury models using suture ligation based on fenestration.Postoperative comparisons were made between the two groups,assessing urination behavior,hematuria,pyuria(infection rate),mortality,scoliosis rate and Basso-Beattie-Bresnahan locomotor rating scale scores at 1,3,5,and 7 days after modeling. RESULTS AND CONCLUSION:Compared with the conventional modeling method,the modified ligation method based on the compression method resulted in faster recovery of urination behavior,lower hematuria rate,lower infection rate,lower mortality rate,lower scoliosis rate,and more concentrated and stable Basso-Beattie-Bresnahan scores(all below 2 points within 1 week).This proves that the modified ligation method based on compression is more suitable for the establishment of spinal cord injury models in rats.
2.Unveiling the molecular features and diagnosis and treatment prospects of immunothrombosis via integrated bioinformatics analysis.
Yafen WANG ; Xiaoshuang WU ; Zhixin LIU ; Xinlei LI ; Yaozhen CHEN ; Ning AN ; Xingbin HU
Chinese Journal of Cellular and Molecular Immunology 2025;41(3):228-235
Objective To investigate the common molecular features of immunothrombosis, thus enhancing the comprehension of thrombosis triggered by immune and inflammatory responses and offering crucial insights for identifying potential diagnostic and therapeutic targets. Methods Differential gene expression analysis and functional enrichment analysis were conducted on datasets of systemic lupus erythematosus (SLE) and venous thromboembolism (VTE). The intersection of differentially expressed genes in SLE and VTE with those of neutrophil extracellular traps (NET) yielded cross-talk genes (CG) for SLE-NET and VTE-NET interaction. Further analysis included functional enrichment and protein-protein interaction (PPI) network assessments of these CG to identify hub genes. Venn diagrams and receiver operating characteristic (ROC) curve analysis were employed to pinpoint the most effective shared diagnostic CG, which were validated using a graft-versus-host disease (GVHD) dataset. Results Differential expression genes in SLE and VTE were associated with distinct biological processes, whereas SLE-NET-CG and VTE-NET-CG were implicated in pathways related to leukocyte migration, inflammatory response, and immune response. Through PPI network analysis, several hub genes were identified, with matrix metalloproteinase 9 (MMP9) and S100 calcium-binding protein A12 (S100A12) emerging as the best shared diagnostic CG for SLE (AUC: 0.936 and 0.832) and VTE (AUC: 0.719 and 0.759). Notably, MMP9 exhibited good diagnostic performance in the GVHD dataset (AUC: 0.696). Conclusion This study unveils the common molecular features of SLE, VTE, and NET, emphasizing MMP9 and S100A12 as the optimal shared diagnostic CG, thus providing valuable evidence for the diagnosis and therapeutic strategies related to immunothrombosis. Additionally, the expression of MMP9 in GVHD highlights its critical role in the risk of VTE associated with immune system disorders.
Humans
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Computational Biology/methods*
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Lupus Erythematosus, Systemic/immunology*
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Protein Interaction Maps/genetics*
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Venous Thromboembolism/therapy*
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Matrix Metalloproteinase 9/genetics*
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Extracellular Traps/metabolism*
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Gene Regulatory Networks
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Thrombosis/immunology*
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Graft vs Host Disease/genetics*
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Gene Expression Profiling
3.Study on the synergistic effects and mechanisms of IL-25 and house dust mite in promoting eosinophilic asthma allergy
Lina Xu ; Xiaoshuang He ; Dong Liu
Acta Universitatis Medicinalis Anhui 2025;60(4):649-655
Objective :
To explore the central role of interleukin(IL)-25 in allergy of eosinophil asthma.
Methods :
Forty 4-5 week-old C57BL/6 mice were used in this study(n=10). Mice were randomly divided into 4 groups: WT+shRNA NT group, WT+HDM group, IL-25 shRNA+IL-25 group, and IL-25 shRNA+IL-25+HDM group. According to each group′s treatment requirement, mice were treated with house dust mite and/or IL-25 by intranasal infusion. HE staining and PAS staining were used for lung tissue section staining and pathological analysis. The levels of type 2 cytokines in bronchoalveolar lavage fluid(BALF) were determined by ELISA. Flow cytometry(FCM) was used to determine the number of type 2 helper T(Th2) cells, the group 2 innate lymphoid cells(ILC2) and eosinophils in lung tissues. Mice were treated with FITC-labeled DQ-ovalbumin(DQ-OVA) by intranasal infusion to determine the antigen presentation ability of eosinophils infiltrated in the lungs.
Results :
HE staining and PAS staining results showed that, compared with WT+shRNA NT group, a large amount of mucus and a large number of eosinophil infiltration were found in the lungs, and subcutaneous thickening of inflammatory airway were observed in the pulmonary vessels, alveolar ducts and the whole alveoli in IL-25 shRNA+IL-25+HDM group. In the IL-25 shRNA+IL-25 group, only a small amount of mucus and a small amount of eosinophil infiltration were found in the lungs, and a small amount of subcutaneous thickening of inflammatory airway were observed in pulmonary vessels, alveolar ducts and the whole alveoli. There was no significant difference in WT+HDM group. Compared with WT+shRNA NT group, the levels of IL-25, IL-4,IL-5,IL-13, IL-33 and interferon-γ(IFN-γ) in BALF significantly increased; the levels of infiltrated Th2, ILC2 and eosinophils in the lung significantly increased; the fluorescence levels of FITC-labeled DQ-OVA in eosinophils infiltrated in lungs significantly increased in IL-25 shRNA+IL-25+HDM group(P<0.05). IL-25 shRNA+IL-25 group mice also showed an increase in the above measurements, but the amplitude were limited(P<0.05). WT+HDM group had no significant difference.
Conclusion
IL-25 plays an essential role in the allergy and eosinophil antigen presentation processes in eosinophil asthma mice.
4.Research on quantitative evaluation of revise requirements of Specifications of Air Sampling for Hazardous Substances Monitoring in the Workplace (GBZ 159-2004)
Tuo LIU ; Xiaoshuang XIE ; Yang LU ; Peng QIN ; Wenjie LI ; Qiuhong ZHU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(2):90-95
Objective:To conduct quantitative evaluation on the revise requirements of Specifications of Air Sampling for Hazardous Substances Monitoring in the Workplace (GBZ 159-2004) , clarify the problems and suggestions during its implementation for improvement, and provide a basis for the revision of the standard.Methods:From April to September 2021, stratified convenient sampling method was adopted and semi-open questionnaire was used to investigate the occupational health personnel in CDC, occupational prevention and control institutes, employers, third-party technical service institutions and universitie. The entropy weight of each index and the score based on entropy weight of GBZ 159 were calculated. Spearman rank correlation analysis was used to describe the correlation between the two indexes and radar chart was drawn for comprehensive evaluation.Results:A total of 151 questionnaires were received from the respondents, of which 147 were valid, with an effective recovery rate of 97.35%, involving 29 provinces, autonomous regions and municipalities. The median G scores of the necessity and urgency of GBZ 159 revision based on entropy weight were 2.84 and 3.17, respectively, and the difference was statistically significant ( M=-25.50, P<0.001) . The trend of the score G of necessity and urgency based on entropy weight was basically the same for all secondary items ( rs=0.9998, P<0.001) , and the score G of urgency based on entropy weight was higher than that of necessity. The highest score G of necessity and urgency based on entropy weight was "3.13 long time sampling", which were 7.56 and 8.23 respectively. This was followed by "3.12 short time sampling", which were 7.19 and 7.13 respectively. Conclusion:GBZ 159 has encountered some new problems and challenges in the implementation process, and some of its technical indicators have been out of line with the actual practice of occupational health at present. These are the two items that urgently needs to be revised and improved, such as "3.13 long time sampling" and "3.12 short time sampling" and other items need to be revised and improved.
5.Study on the implementation effect evaluation of Specifications of Air Sampling for Hazardous Substances Monitoring in the Workplace (GBZ 159-2004)
Tuo LIU ; Xiaoshuang XIE ; Yang LU ; Peng QIN ; Wenjie LI ; Qiuhong ZHU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(2):136-140
Systematically evaluate the implementation of Specifications of Air Sampling for Hazardous Substances Monitoring (GBZ 159-2004) , so as to provide technical basis for the future standard revision. The semi-structured interview method was used to interview the industry experts from centers for disease control and prevention, occupational disease prevention and control hospitals/institutes, employers, third-party technical service institutions and universities, and the induction method was used to refine the topics and relevant suggestions. Some technical indicators of GBZ 159 are not suitable for the current actual work of occupational health, and need to be revised and improved urgently. The revised GBZ 159 should comply with the current situation of China's industrial development and the development of occupational health testing equipment in the new era, and improve the relevant technical requirements of sampling quality control.
6.Regulation of chondrocyte autophagy by acupotomy to promote chondrocyte homeostasis in osteoarthritis
Xiaofei JIA ; Li RAN ; Xiaoshuang MA ; Xiaoyan HEI ; Jiani LIU ; Nan YANG ; Haibin MA ; Jingpeng CHANG
Chinese Journal of Tissue Engineering Research 2024;28(34):5452-5457
BACKGROUND:Acupotomy is an effective method for the clinical treatment of osteoarthritis,with affirmed clinical outcomes,but the specific mechanisms remain unclear OBJECTIVE:To investigate the role of acupotomy in modulating chondrocyte autophagy to promote chondrocyte homeostasis in osteoarthritis. METHODS:Twenty-eight New Zealand rabbits were randomly divided into control group,osteoarthritis group,acupotomy group,and hyaluronic acid group,with seven rabbits in each group.The knee osteoarthritis rabbit model was prepared using the Videman method in the latter three groups.After modeling,the control group and osteoarthritis group received no interventions.The acupotomy group received acupotomy treatment 15 minutes per time,once a week,while the hyaluronic acid group received intra-articular injection of hyaluronic acid once a week,with a continuous treatment duration of 5 weeks.The day after the final intervention,knee joint macrostructure was observed using DR imaging,chondrocyte ultrastructure was examined through transmission electron microscopy,apoptosis of chondrocytes was assessed using Tunel staining,and western blot analysis was used to detect the expression of proteins related to the PI3K/Akt/mTOR pathway. RESULTS AND CONCLUSION:The DR imaging results revealed that the osteoarthritis group exhibited narrowed knee joint spaces and the formation of periarticular osteophytes,while the hyaluronic acid group and acupotomy group showed widened knee joint spaces with a reduction in periarticular osteophytes.Transmission electron microscopy results demonstrated a decreased number of autophagosomes in chondrocytes in the osteoarthritis group,along with nuclear shrinkage,nuclear membrane rupture,incomplete organelle morphology,and a clear tendency towards cell death.In contrast,both the hyaluronic acid group and acupotomy group exhibited a significant increase in autophagosomes,intact nuclear membranes,and a well-preserved cellular state.Tunel staining results indicated a considerable decrease in the number of apoptotic cells in the hyaluronic acid group and acupotomy group compared with the osteoarthritis group.Western blot results revealed that,compared with the control group,the expression levels of Beclin1,Cath D,and LC3II/LC3I were significantly decreased in the osteoarthritis group(P<0.05),while the expression levels of p-Akt/Akt and p-mTOR/mTOR were significantly increased(P<0.05);compared with the osteoarthritis group,the expression levels of Beclin1,Cath D,and LC3II/LC3I were significantly increased in both the hyaluronic acid group and acupotomy group(P<0.05),while the expression levels of p-Akt/Akt and p-mTOR/mTOR were significantly decreased(P<0.05).To conclude,acupotomy intervention can modulate the PI3K/Akt/mTOR signaling pathway to enhance the autophagic level in chondrocytes,thereby maintaining chondrocyte homeostasis.This ultimately leads to a slowdown in cartilage degeneration.
7.Influence of geometric factor on detection efficiency in low-background α/β radioactivity measurement
Chinese Journal of Radiological Health 2024;33(2):183-188
Objective To study the influence of the radius of the active zone of the plane source and the horizontal distance between the source center and the center of the detection window on the detection efficiency of low-background α/β measuring instruments. Methods The geometric factor was calculated under different conditions in MATLAB, and a traceable plane source was used to test and analyze two commonly used measuring instruments. Results With the other parameters unchanged, when the radius of the active zone R1 = 9 mm, the G value was greatest; and when the horizontal distance a = 0 mm, the G value was greatest. According to the calculation formula of the verification regulation, for the 90Sr-90Y planar source, at the maximum value of a, the detection efficiency of FYFS-400X was 1.2% lower, and that of MPC-9604 was 3.2% lower; for the 241Am planar source, the detection efficiency of MPC-9604 was 1.9% lower at the maximum value of a. Conclusion The geometric factor decreases with the increase in the radius of the active zone and the horizontal distance between the source center and the center of the detection window. At the central position, the geometric factor has less influence on the relative standard uncertainty of detection efficiency. When using a standard plane source for counting, measures should be taken to ensure that the plane source is always centrally located in the sample tray.
8.The mechanism of N-acetylserotonin regulating microglial polarization via NOD1/Rip2 pathway in rats after retinal ischemia reperfusion
Ying XU ; Jianliang LIU ; Yuze ZHAO ; Chenxu WANG ; Xinhao FU ; Xiaoshuang LI ; Xiaoli WANG ; Yansong ZHAO
Chinese Journal of Ocular Fundus Diseases 2024;40(4):287-295
Objective:To investigate the effect of N-acetylserotonin (NAS) on the retinal microglia polarization in retinal ischemia-reperfusion injury (RIRI) rats and explore its mechanism via nucleotide-bound oligomeric domain 1 (NOD1)/receptor interacting protein 2 (Rip2) pathway.Methods:Healthy male Sprague Dawley rats were randomly divided into Sham ( n=21), RIRI ( n=21) and NAS (injected intraperitoneally 30 min before and after modeling with NAS, 10 mg/kg, n=18) groups, using random number table. And the right eye was used experimental eye. The RIRI model of rats in RIRI group and NAS group was established by anterior chamber high intraocular pressure method. Rats in NAS group were intraperitoneally injected with 10 mg/kg NAS before and 30 min after modeling, respectively. The retinal morphology and the number of retinal ganglion cell (RGC) in each group were detected by hematoxylin-eosin staining and immunohistochemical staining. The effect of NAS on polarization of retinal microglia was detected by immunofluorescence staining. Transcriptome sequencing technology was used to screen out the differentially expressed genes between Sham and RIRI groups. Western blot and real-time quantitative polymerase chain reaction (RT-PCR) were used to examine the differentially expressed genes. Immunohistochemical staining, Western blot and RT-PCR were used to investigate the effect of NAS on the expression of NOD1 and Rip2 protein and mRNA in retinal tissue and microglia of rats. General linear regression analysis was performed to determine the correlation between the number difference of NOD1 + cells and the number difference of M1 and M2 microglia in retinal tissues of rats in NAS group and RIRI group. Results:A large number of RGC were observed in the retina of rats in Sham group. 24 h after modeling, compared with Sham group, the inner retinal thickness of rats in RIRI group was significantly increased and the number of RGC was significantly decreased. The thickness of inner retina in NAS group was significantly thinner and the number of RGC was significantly increased. Compared with Sham group, the number of retinal microglia of M1 and M2 in RIRI group was significantly increased. Compared with RIRI group, the number of M1 microglia decreased significantly and the number of M2 microglia increased significantly in NAS group. There was statistical significance in the number of M1 and M2 microglia in the retina of the three groups ( P<0.05). Transcriptome sequencing results showed that retinal NOD1 and Rip2 were important differential genes 24 h after modeling. The mRNA and protein relative expressions of NOD1 and Rip2 in retina of RIRI group were significantly higher than those of Sham group, with statistical significance ( P<0.05). The number of NOD1 + and Rip2 + cells and the relative expression of mRNA and protein in retinal microglia in RIRI group were significantly higher than those in Sham group, and NAS group was also significantly higher than that in Sham group, but lower than that in RIRI group, with statistical significance ( P<0.05). The number of Iba-1 +/NOD1 + and Iba-1 +/Rip2 + cells in retinal microglia in RIRI group was significantly increased compared with that in Sham group, and the number of Iba-1 +/Rip2 + cells in NAS group was significantly decreased compared with that in RIRI group, but still significantly higher than that in Sham group, with statistical significance ( P<0.05). Correlation analysis results showed that the difference of retinal NOD1 + and Rip2 + cells in NAS group and RIRI group was positively correlated with that of M1 microglia ( r=0.851, 0.895), and negatively correlated with that of M2 microglia ( r=-0.797, -0.819). The differences were statistically significant ( P<0.05). Conclusion:NAS can regulate the microglial polarization from M1 to M2 phenotype, the mechanism is correlated with the NOD1/Rip2 pathway.
9.Effect of 30% supramolecular salicylic acid combined with 420 nm intense pulsed light on facial seborrheic dermatitis
Ying DENG ; Hongqiu YANG ; Xiaoshuang DU ; Ping LIU ; Yu DU
Chinese Journal of Medical Aesthetics and Cosmetology 2024;30(4):377-381
Objective:To investigate the clinical efficacy of 30% supramolecular salicylic acid combined with 420 nm intense pulsed light in the treatment of facial seborrheic dermatitis.Methods:A total of 60 patients with facial seborrheic dermatitis were selected from January 2018 to December 2021, that were randomly divided into 30% salicylic acid group, 420 nm intense pulsed light group and combined group. Each group had 20 patients. They were treated once every two weeks in four consecutive rounds. The adverse reactions of patients were followed up each time, and the data were analysed and evaluated four weeks after the last treatment.Results:After treatment, the total score of clinical symptoms and signs in the combined group were significantly lower than those in the other groups (3.95±1.731 vs. 7.15±2.059, 7.50±1.732, F=22.434, P<0.05). The effective rate in combined group was 95%, which was significantly higher than 65% of salicylic acid group and 60% of intense pulsed light group (χ 2=7.33, P<0.05), and no obvious adverse reactions were found in the three groups. Conclusions:30% supramolecular salicylic acid combined with 420 nm intense pulsed light is safe and effective in the treatment of facial seborrheic dermatitis.
10.Correlation between non-high density lipoprotein cholesterol,homocysteine levels and cognitive impairment,prognosis in patients with thalamic infarction
Xiaoshuang LIU ; Ning GUO ; Wanyu ZHAO
Journal of Xinxiang Medical College 2024;41(1):43-48
Objective To investigate the correlation between non-high density lipoprotein cholesterol(non-HDL-C),homocysteine(Hey)levels and cognitive impairment,prognosis in patients with thalamic infarction.Methods Eighty thalamic infarction patients admitted to Puyang People's Hospital from March 2017 to March 2021 were selected as the research sub-jects.According to Montreal cognitive assessment(MoCA)score,the patients were divided into cognitive impairment group(MoCA score<26,n=35)and cognitive normal group(MoCA score 26-30,n=45).Another 50 healthy individuals who un-derwent physical examinations during the same period were selected as the control group.The serum cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),Hcy levels of all subjects were detected by fully automated biochemical analyzer,and the non-HDL-C levels were calculated.The correlations between non-HDL-C,Hey levels and MoCA scores were analyzed by Pearson correlation analysis,and the diagnostic efficacy of non-HDL-C,Hcy levels for cognitive impairment of patients with tha-lamic infarction was analyzed by receiver operating characteristic curve.According to the modified Rankin scale(mRS)score,the patients with thalamic infarction were divided into good prognosis group(mRS score ≤2,n=50)and poor prognosis group(mRS score 3-6,n=50).The independent risk factors for poor prognosis of patients with thalamic infarction was analyzed by multivariate logistic regression.Results The serum levels of non-HDL-C and Hcy of patients in the cognitive impairment group were significantly higher than those in the cognitive normal group and control group(P<0.05);there was no significant difference in the serum non-HDL-C,Hcy levels of subjects between the control group and the cognitive normal group(P>0.05).The non-HDL-C,Hcy levels were negatively correlated with MoCA scores in thalamic infarction patients(P<0.05).The area under the curve(AUC)of Hcy level in the diagnosis of cognitive impairment was 0.709,the sensitivity was 0.724 and the specificity was 0.630;the AUC of non-HDL-C level in the diagnosis of cognitive impairment was 0.738,the sensitivity was 0.701 and the specificity was 0.870;the AUC of Hey combined with non-HDL-C in the diagnosis of cognitive impairment was 0.769,the sensitivity was 0.758 and the specificity was 0.889.The diagnostic efficacy of Hey combined with non-HDL-C for cognitive impairment was superior to that of non-HDL-C or Hey alone.Atrial fibrillation,elevated levels of non-HDL-C,Hcy and NIHSS score were independent risk factors for poor prognosis of patients with thalamic infarction(P<0.05).Conclusion The levels of non-HDL-C and Hey are positively correlated with cognitive impairment in patients with thalamic infarction.The levels of non-HDL-C and Hey can be used to diagnose cognitive impairmnent in patients with thalamic infarction,and the combined detection has a better diagnostic effect.Atrial fibrillation,elevated levels of non-HDL-C,Hcy and NIHSS score are independent risk factors for poor prognosis in patients with thalamic infarction.


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