1.NINJ1 impairs the anti-inflammatory function of hUC-MSCs with synergistic IFN-γ and TNF-α stimulation.
Wang HU ; Guomei YANG ; Luoquan AO ; Peixin SHEN ; Mengwei YAO ; Yuchuan YUAN ; Jiaoyue LONG ; Zhan LI ; Xiang XU
Chinese Journal of Traumatology 2025;28(4):276-287
PURPOSE:
To investigate the regulatory role of nerve injury-induced protein 1 (NINJ1) in the anti-inflammatory function of human umbilical cord mesenchymal stem cells (hUC-MSCs) co-stimulated by interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α).
METHODS:
hUC-MSCs were expanded in vitro using standard protocols, with stem cell characteristics confirmed by flow cytometry and multilineage differentiation assays. The immunomodulatory properties and cellular activity of cytokine-co-pretreated hUC-MSCs were systematically evaluated via quantitative reverse transcription RT-qPCR, lymphocyte proliferation suppression assays, and Cell Counting Kit-8 viability tests. Transcriptome sequencing, Western blotting and small interfering RNA interference were integrated to analyze the regulatory mechanisms of NINJ1 expression. Functional roles of NINJ1 in pretreated hUC-MSCs were elucidated through gene silencing combined with lactate dehydrogenase release assays, Annexin V/Propidium Iodide apoptosis analysis, macrophage co-culture models, and cytokine Enzyme-Linked Immunosorbent Assay. Therapeutic efficacy was validated in a cecal ligation and puncture-induced septic mouse model: 80 mice were randomly allocated into 4 experimental groups (n=20/group): sham group (laparotomy without cecal ligation); phosphate-buffered saline-treated group (cecal ligation and puncture (CLP) + 0.1 mL phosphate-buffered saline); hUC-MSCs (small interfering RNA (siRNA)-interferon-gamma and tumor necrosis factor-alpha co-stimulation (IT))-treated group (CLP + hUC-MSCs transfected with scrambled siRNA); and hUC-MSCs (siNINJ1-IT)-treated group (CLP + hUC-MSCs with NINJ1-targeting siRNA).
RESULTS:
hUC-MSCs demonstrated compliance with International Society for Cellular Therapy criteria, confirming their stem cell identity. IFN-γ/TNF-α co-pretreatment enhanced the immunosuppressive capacity of hUC-MSCs, accompanied by the reduction of cellular viability, while concurrently upregulating pro-inflammatory cytokines such as interleukin-6 and interleukin-1β. This co-stimulation significantly elevated NINJ1 expression in hUC-MSCs, whereas genetic silencing of NINJ1 effectively suppressed pro-inflammatory cytokine production and attenuated damage-associated molecular patterns release through inhibition of programmed plasma membrane rupture. Furthermore, the NINJ1 interference potentiated the ability of cytokine-pretreated hUC-MSCs to suppress LPS-induced pro-inflammatory responses in RAW264.7 macrophages. In cecal ligation and puncture-induced sepsis model, NINJ1-silenced hUC-MSCs exhibited enhanced therapeutic efficacy, manifested by reduced systemic inflammation and multi-organ damage.
CONCLUSION
Our findings shed new light on the immunomodulatory functions of cytokine-primed MSCs, offering groundbreaking insights for developing MSC-based therapies against inflammatory diseases via interfering the expression of NINJ1.
Mesenchymal Stem Cells/drug effects*
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Animals
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Interferon-gamma/pharmacology*
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Tumor Necrosis Factor-alpha/pharmacology*
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Humans
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Mice
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Umbilical Cord/cytology*
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Cells, Cultured
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Apoptosis
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Male
3.Sanguinarine alleviates ulcerative colitis in mice by regulating the Nrf2/NF-κB pathway
Na ZHAO ; Mengdi SHEN ; Rui ZHAO ; Di AO ; Zetan LUO ; Yinliang ZHANG ; Zhidong XU ; Fangtian FAN ; Hailun ZHENG
Journal of Southern Medical University 2024;44(8):1467-1475
Objective To investigate the mechanism of sanguinarine(SA)for alleviating ulcerative colitis(UC)induced by dextran sodium sulfate(DSS)in mice.Methods Male C57BL/6 mouse models of 3.5%DSS-induced UC were randomized for treatment with 1,5 and 10 mg/kg SA by gavage,400 mg/kg sulfasalazine by gavage,or 10 mg/kg SA combined with intraperitoneal injection of 30 mg/kg ML385(a Nrf2 inhibitor).The changes in intestinal inflammation was assessed by monitoring weight changes,disease activity index(DAI)score,colon length measurement,and HE staining.After the treatments,the colon tissues were collected for detection of malondialdehyde(MDA)content using colorimetry,mRNA expressions of inflammatory factors using RT-qPCR,and the expressions of Nrf2,HO-1,Keap-1,p-p65,p65,occludin,and ZO-1 proteins were detected using Western blotting.Results SA treatment obviously alleviated weight loss,colon length shortening and DAI score increase and ameliorated structural destruction of the colon glands and colonic crypts in mice with DSS-induced UC.SA intervention significantly decreased the levels of TNF-α,IL-1β and IL-6 mRNA and lowered ROS and MDA levels in the colon tissue of UC mice.The mouse models receiving SA treatment showed significantly increased expressions of Nrf2,HO-1,occludin and ZO-1 and lowered expressions of Keap-1 and P-P65 in the colon tissue without significant changes of p65 expression,and these changes were SA dose-dependent.Treatment with ML385 obviously attenuated the effect of high-dose SA for improving UC in the mouse models.Conclusion SA can improve UC-like enteritis in mice possibly by activating the Nrf2 pathway and inhibiting the NF-κB pathway in the colon tissue.
4.Sanguinarine alleviates ulcerative colitis in mice by regulating the Nrf2/NF-κB pathway
Na ZHAO ; Mengdi SHEN ; Rui ZHAO ; Di AO ; Zetan LUO ; Yinliang ZHANG ; Zhidong XU ; Fangtian FAN ; Hailun ZHENG
Journal of Southern Medical University 2024;44(8):1467-1475
Objective To investigate the mechanism of sanguinarine(SA)for alleviating ulcerative colitis(UC)induced by dextran sodium sulfate(DSS)in mice.Methods Male C57BL/6 mouse models of 3.5%DSS-induced UC were randomized for treatment with 1,5 and 10 mg/kg SA by gavage,400 mg/kg sulfasalazine by gavage,or 10 mg/kg SA combined with intraperitoneal injection of 30 mg/kg ML385(a Nrf2 inhibitor).The changes in intestinal inflammation was assessed by monitoring weight changes,disease activity index(DAI)score,colon length measurement,and HE staining.After the treatments,the colon tissues were collected for detection of malondialdehyde(MDA)content using colorimetry,mRNA expressions of inflammatory factors using RT-qPCR,and the expressions of Nrf2,HO-1,Keap-1,p-p65,p65,occludin,and ZO-1 proteins were detected using Western blotting.Results SA treatment obviously alleviated weight loss,colon length shortening and DAI score increase and ameliorated structural destruction of the colon glands and colonic crypts in mice with DSS-induced UC.SA intervention significantly decreased the levels of TNF-α,IL-1β and IL-6 mRNA and lowered ROS and MDA levels in the colon tissue of UC mice.The mouse models receiving SA treatment showed significantly increased expressions of Nrf2,HO-1,occludin and ZO-1 and lowered expressions of Keap-1 and P-P65 in the colon tissue without significant changes of p65 expression,and these changes were SA dose-dependent.Treatment with ML385 obviously attenuated the effect of high-dose SA for improving UC in the mouse models.Conclusion SA can improve UC-like enteritis in mice possibly by activating the Nrf2 pathway and inhibiting the NF-κB pathway in the colon tissue.
5.Efficacy assessment for NMES in improving muscle strength in patients with SAP complicated by ARDS
Dingrong FAN ; Hengyu ZHOU ; Ying CAI ; Botao TAN ; Qianqian WANG ; Feng ZHOU ; Xiaoyun RAN ; Xiaodong CHEN ; Ao SHEN
Journal of Army Medical University 2024;46(22):2539-2546
Objective To evaluates the impact of early application of neuromuscular electrical stimulation(NMES)on muscle strength,clinical outcomes,and long-term quality of life improvements in patients with severe acute pancreatitis(SAP)complicated with acute respiratory distress syndrome(ARDS).Methods A total of 75 patients diagnosed with SAP and ARDS admitted in Department of Critical Care Medicine of our hospital from September 2022 to August 2023 were recruited and then randomly divided into NMES group(n=37)and control group(n=38).After 16 patients were excluded,including 8 died during treatment,3 discharged and 5 received palliative care,there were finally 29 patients in the NMES group and 30 in the control group.Within 48 h after ICU admission,the NMES group received NMES 1 h per day,for 7 d in addition to standard rehabilitation intervention.While,the control group were given conventional interventions for rehabilitation.Assessments at baseline and post-treatment included the incidence of ICU-acquired weakness(ICU-AW),Medical Research Council(MRC)score,duration of mechanical ventilation,lengths of ICU and total hospital stays,and activity,thickness and thickening fraction of the diaphragm.Mortality rates and Barthel index(BI)for self-care ability in 1,3 and 6 months after discharge were recorded for follow-up assessments.Results The NMES group had significantly lower incidence of ICU-AW(P<0.05),higher upper and lower limb MRC scores and overall MRC score at ICU discharge(P<0.05),shorter durations of mechanical ventilation,ICU stay,and total hospital stay when compared with the control group(P<0.05).There was no statistical difference in the BI at 1 month post-discharge between the 2 groups,but the indexes at 3 and 6 months were notably higher in the NMES group than the control group(P<0.05).No obvious differences were observed between the 2 groups in terms of diaphragm activity,thickness,or thickening scores at enrollment,ICU discharge,or hospital discharge,nor in mortality rates at 1,3,and 6 months after discharge.Conclusion Combined NMES and early rehabilitation therapy can improve muscle strength and reduce length of hospital stay in SAP patients complicated with ARDS,and may enhance long-term quality of life.However,it does not significantly affect diaphragm function or mortality rates.
6.Application and effective evaluation of simulation teaching of ultrasound in education of critical care nursing
Yanling SHEN ; Ying ZHANG ; Cheng GONG ; Zhen LI ; Ao ZHANG
China Medical Equipment 2024;21(6):178-182
Objective:To explore the application effect of simulation teaching of ultrasound in education of critical care nursing.Methods:From January to December 2023,a total of 40 nurses who received advanced training in Critical Care Medicine Department of China-Japan Friendship Hospital were selected,and they were divided into observation group(20 cases)and control group(20 cases)by random number table method.The observation group adopted simulation teaching method of ultrasound based on the platform of American patriot comprehensive ultrasound virtual training system,and the control group adopted conventional ultrasound teaching method.The theoretical course teaching,the examination results of practical operation and teaching satisfaction between the two groups were compared.Results:The operation assessment,homework after class and total score of assessment of observation group were respectively(27.9±1.91)points,(16.25±1.33)points and(77.75±4.12)points,which were significantly higher than those of control group,the differences were statistically significant(t=13.375,3.635,8.814,P<0.05),respectively.The average scores of lung ultrasound,assessment of gastric residual volume,assessment of skin pressure injury and arteriovenous puncture of examination achievements in observation group were respectively(5.30±1.21)points,(5.85±0.81)points,(5.10±0.91)points and(6.05±0.89)points,which were higher than those in control group,respectively,and the differences were statistically significant(t=5.792,5.264,5.832,5.620,P<0.05).The total scores of teaching arrangement,teaching effect,overall harvest and satisfaction of nurses in observation group were respectively(22.75±1.30)points,(23.90±1.02)points,(24.00±1.30)points and(93.05±3.66)points,which were significantly higher than those in control group,and the differences were statistically significant(t=6.564,8.860,6.694,7.441,P<0.05),respectively.Conclusion:Simulation teaching of ultrasound can significantly improve the operation ability of nurses who receive advanced training in the teaching of critical care nursing,as well as the mastery of teaching content and learning interest of them,which can enhance the quality of nursing clinical teaching and the satisfaction of teaching.
7.Visual analysis of the research hotspots and trends of emergency nursing training at home and abroad based on CiteSpace
Xinyu DUAN ; Hongzhen XIE ; Ao WU ; Tenggang SHEN ; Wenjuan XU ; Qiaoqiao ZHANG ; Qunfang YANG ; Duo LIU ; Xiaoqi FAN ; Jianjing WANG
Chinese Journal of Practical Nursing 2024;40(26):2059-2068
Objective:To analyze the status quo, hotspots and fronts of emergency nursing training research at home and abroad in the past ten years, and to provide reference and ideas for the efficient development of emergency nursing training in China.Methods:CiteSpace 6.2.R2 software was used to visually analyze the Chinese and English literatures on emergency nursing training included in CNKI and Web of Science core databases from January 1, 2013 to June 1, 2023.Results:A total of 1 177 Chinese literatures and 1 163 English literatures were included. The number of foreign articles in this field increased year by year, while the number of domestic articles showed a downward trend since 2018. There were many stable core author groups and core institution groups in foreign countries, while there was less cooperation among domestic authors and institutions. The common research hotspots and frontiers at home and abroad focused on broadening the training audience of emergency nursing, innovating the training methods of emergency nursing, strengthening the evaluation of the effect of emergency nursing training, and paying attention to the training experience and needs of nurses. Foreign researches also focused on specialized nurses, interprofessional education and nurses′mental health, etc, and the research direction was diversified.Conclusions:The development stages of emergency nursing training researches at home and abroad are different, and the research hotspots are different. In the future, we should learn from foreign research, strengthen interdisciplinary cooperation, improve the depth and breadth of research, and strengthen the cooperation between authors, institutions and countries to promote the high-quality development of emergency nursing training research in China.
8.Effect of Xin-Gui Gel Plaster on Peripheral Neuropathy in Diabetic Rats
Xiao TAN ; Huiwen WANG ; Ao SHEN ; Yanfeng XIU ; Haiying WANG
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(1):77-82
Objective To observe the effects of Xin-Gui Gel Plaster(Cinnamomi Cortex,Asari Radix et Rhizoma,Euodiae Fructus,Chuanxiong Rhizoma,Borneolum Syntheticum)on peripheral neuropathy in diabetic rats.Methods A single intraperitoneal injection of 1%streptozotonic(STZ,35 mg·kg-1)was used to replicate a type 2 diabetes mellitus(T2DM)rat model followed by the induction of diabetic peripheral neuropathy(DPN)in combination with a long-term(8 consecutive weeks)high-fat and high-sugar diet.SD rats were randomly divided into normal group,model group,Mecobalamin group and Xin-Gui Gel Plaster group,with 6 rats in each group.The rats in the Xin-Gui Gel Plaster group were given Xin-Gui Gel Plaster at acupoints once a day for 8 weeks;the rats in the Mecobalamin group were given Mecobalamin solution by gavage(0.045 mg·kg-1),and the rats in the normal group and the model group were given physiological saline by gavage.Body mass and fasting blood glucose(FBG)were measured at weeks 2,4,6 and 8;the latency of thermal withdrawal latency(TWL)was measured at weeks 4 and 8;nerve conduction velocities,including motor nerve conduction velocity(MNCV)and sensory nerve conduction velocity(SNCV),were measured at week 8;and serum total cholesterol(TC),triglyceride(TG),fasting blood glucose(FBG)were measured using a fully automated biochemical analyzer.Fasting insulin(FINS)levels were detected by ELISA,and the index of insulin resistance(HOMA-IR)was calculated;and pathological changes in the sciatic nerve tissues of rats were observed by HE staining.Results Compared with the normal group,rats in the model group had significantly lower body mass and FINS levels(P<0.01),significantly higher levels of FBG,TC,TG and HOMA-IR(P<0.05,P<0.01);TWL,MNCV and SNCV were significantly decreased(P<0.01),and sciatic nerve fibres were disorganized and loosely aligned,with demyelination,axon atrophy and vacuole-like phenomenon.Compared with the model group,there was no significant change in body mass and levels of FBG,TC and TG in the Xin-Gui Gel Plaster group(P>0.05),FINS level was significantly increased(P<0.05),and HOMA-IR levels was significantly decreased(P<0.05);TWL,MNCV and SNCV in the Mecobalamin group and Xin-Gui Gel Plaster group were significantly increased(P<0.05,P<0.01),sciatic nerve lesions were improved to different degrees,nerve fibre arrangement was more regular,myelin deficiency and axonal atrophy were significantly improved.Conclusion Xin-Gui Gel Plaster can improve insulin resistance,relieve thermal stimulation sensitivity,improve sciatic nerve conduction velocity to a certain extent in DPN rats,and have a protective effect on peripheral nerves in diabetic rats,but the hypoglycemic and hypolipidemic effects are not obvious.
9.Chemical composition analysis and value evaluation of stems and leaves of Astragalus membranaceus var. mongholicus.
Qiang-Xiong WANG ; Sheng GUO ; Ke-Xin SHEN ; Hui-Wei LI ; Hao-Kuan ZHANG ; Yi-Jun XIE ; Er-Xin SHANG ; Jin-Ao DUAN
China Journal of Chinese Materia Medica 2023;48(24):6600-6612
This study aimed to provide data support for resource utilization of the stems and leaves of Astragalus membranaceus var. mongholicus(SLAM) by analyzing and evaluating the chemical constituents. The crude protein, crude fiber, and soluble saccharide of SLAM were analyzed by Kjeldahl method, filtration method, and UV-Vis spectrophotometry, respectively. The nucleosides, amino acids, flavonoids, and saponins of SLAM were analyzed by ultraperformance liquid chromatography-triple quadrupole mass spectrometry(UPLC-TQ-MS). Combined with principal component analysis(PCA), the quality difference of resource components of SLAM was comprehensively evaluated. The results showed that the average content of crude protein, crude fiber, total polysaccharide, and redu-cing sugar in SLAM was 5.11%, 30.33%, 11.03 mg·g~(-1), and 31.90 mg·g~(-1), respectively. Six nucleosides, 15 amino acids, 22 flavonoids, and one saponin were detected, with an average content of 1.49 mg·g~(-1), 6.00 mg·g~(-1), 1.86 mg·g~(-1), and 35.67 μg·g~(-1), respectively. The content of various types of chemical components in SLAM differed greatly in different harvesting periods and growing years. The results of PCA showed that the quality of SLAM produced in Ningxia was superior. The results can provide references for the utilization of SLAM.
Astragalus propinquus/chemistry*
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Gas Chromatography-Mass Spectrometry
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Flavonoids/analysis*
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Plant Leaves/chemistry*
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Amino Acids
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Saponins/analysis*
10.Research status of screening and evaluation tool in pediatric delirium
Xiao ZHANG ; Min SHEN ; Hui CHENG ; Ying WANG ; Lijuan AO
Chinese Pediatric Emergency Medicine 2023;30(9):665-670
Early identification and intervention is very important in pediatric delirium.The pediatric confusion assessment method for the ICU and the cornell assessment of pediatric delirium are recommended to screen and evaluate pediatric delirium.We can also choose the appropriate tool according to children′s age, anesthesia status and drug use.Objective evaluation tool has been investigated as well, including biomarkers, imaging studies, electroencephalogram and so on.Electroencephalogram has some typical features, which is related to the occurrence of delirium.For the further research, we should look deep into quantitative electroencephalogram and multimodality model to evaluate and predict delirium.

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