1.Shashen Maidong Tang Enhances Efficacy of Chemotherapy in Mouse Model of Lewis Lung Cancer by Modulating JAK2/STAT3 Signaling Pathway
Lin YU ; Yaoyao WANG ; Limin LIU ; Zuowei HU ; Yanping ZHOU ; Shang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):1-10
ObjectiveTo predict the mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer via network pharmacology and validate the prediction results in animal experiments. MethodsThe potential mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer was predicted by network pharmacology, liquid chromatography-mass spectrometry (LC-MS), and molecular docking methods. C57/BL6 mice were assigned into normal, model, cisplatin, and Shasheng Maidong Tang+cisplatin groups. In addition to the normal group, the remaining groups were injected subcutaneously with 0.2 mL of 1×107 cells·mL-1 Lewis lung cancer cells to establish the Lewis lung cancer model. The daily gavage dose of Shasheng Maidong Tang was 3.58 g·kg-1, and the concentration of cisplatin intraperitoneally injected on every other day was 2 mg·kg-1. Drugs were administered for 14 d. The changes in the tumor volume and the rate of tumor suppression were monitored, and the tumor histopathological changes were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay was employed to measure the interleukin (IL)-6 and interferon (IFN)-γ levels in peripheral blood. Real-time PCR was performed to quantify the mRNA levels of Janus kinase 2 (JAK2), signal transducer and activator of transcription 1 (STAT1), and signal transducer and activator of transcription 3 (STAT3) in the tumor tissue of mice. Western blot was employed to determine the protein levels of JAK2, STAT3, B-cell lymphoma-2 (Bcl-2), cysteinyl aspartate-specific proteinase-3 (Caspase-3), and Pim-1 proto1 (PIM1) in the tumor tissue. Immunohistochemistry was employed to detect the expression of Bcl-2 and PIM1 in the tumor tissue. ResultsNetwork pharmacological predictions indicated that Shasheng Maidong Tang might enhance the efficacy of chemotherapy for lung cancer by regulating nitrogen metabolism, AGE-RAGE signaling pathway, cancer pathway, and JAK/STAT signaling pathway. The experimental results demonstrated that tumor volume in the cisplatin group and Shasheng Maidong Tang+cisplatin group was reduced compared with the model group, with statistically distinct differences observed on days 14, 17, 20 post modeling (P<0.05). Notably, the Shasheng Maidong Tang+cisplatin therapy further decreased tumor volume compared with the cisplatin group, showing marked reductions on days 17 and 20 (P<0.05), consistent with trends visualized in tumor volume comparison charts. The Shasheng Maidong Tang+cisplatin group exhibited higher tumor inhibition rate than the cisplatin group (P<0.05). Histopathological analysis via HE staining revealed that the tumors in the model group displayed frequent nuclear mitosis, densely arranged cells, hyperchromatic nuclei, and no necrosis. Cisplatin treatment induced partial necrosis and vacuolization, while the Shasheng Maidong Tang+cisplatin group exhibited extensive necrotic regions, maximal vacuolization, disarranged tumor cells, and minimal mitotic activity. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed elevated level of IFN-γ (P<0.01) and declined level of IL-6 (P<0.01) in the peripheral blood. Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented elevated level of IFN-γ (P<0.01) and lowered level of IL-6 (P<0.01) in the peripheral blood. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin groups showed down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level STAT1 (P<0.01). Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level of STAT1 (P<0.01). Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), and STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). Compared with the cisplatin group, Shasheng Maidong Tang+cisplatin group presented down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). The Bcl-2 and PIM1 expression results obtained by immunohistochemistry were consistent with those of Western blot. ConclusionShasheng Maidong Tang may enhance the efficacy of chemotherapy in the mouse model of Lewis lung cancer by regulating the JAK2/STAT3 signaling pathway.
2.Structural insights into the distinct ligand recognition and signaling of the chemerin receptors CMKLR1 and GPR1.
Xiaowen LIN ; Lechen ZHAO ; Heng CAI ; Xiaohua CHANG ; Yuxuan TANG ; Tianyu LUO ; Mengdan WU ; Cuiying YI ; Limin MA ; Xiaojing CHU ; Shuo HAN ; Qiang ZHAO ; Beili WU ; Maozhou HE ; Ya ZHU
Protein & Cell 2025;16(5):381-385
3.Improvement effect of Xuebijing injection on blood-brain barrier damage in mice with anti-NMDAR encephalitis and its regulatory effect on Th17/Treg imbalance
Chaosheng ZENG ; Lin CHEN ; Limin YAN ; Huaijie XING ; Li LI ; Shaozhu HUANG ; Min CHEN ; Yong CHANG ; Bing KUANG ; Xiaoyan LI
Journal of Jilin University(Medicine Edition) 2025;51(5):1211-1220
Objective:To investigate the effect of Xuebijing injection against blood-brain barrier(BBB)damage in the mice with anti-N-methyl-D-aspartate receptor(NMDAR)encephalitis,and to elucidate its regulatory effect on the imbalance of helper T cells 17(Th17)/regulatory T cells(Treg).Methods:The active immunization models of anti-NMDAR encephalitis in the mice were established using glutamate receptor N1 subunit(GluN1)356-385 antigen peptide,and the serum anti-NMDAR immunoglobulin G(IgG)antibody levels were detected by enzyme-linked immunosorbent assay(ELISA).The healthy mice without modeling were served as control group,and the mice with successful modeling were randomly divided into model group,low dose of Xuebijing injection(XBJ-L)group,and high dose of Xuebijing injection(XBJ-H)group,with 10 mice in each group.After modeling,the mice in XBJ-L and XBJ-H groups were intraperitoneally injected with 5 and 10 mL·kg-1 Xuebijing injection,respectively.The Longa score was used to assess the neurological impairment of the mice in various groups;evans blue(EB)staining was used to determine the BBB permeability;immunofluorescence staining was used to detect the expressions of zonula occludens 1(ZO-1)and Occludin in cerebral cortex of the mice in various groups;Western blotting method was used to determine the expression levels of ZO-1,Occludin,Claudin-5,and neuron-specific nuclear protein(NeuN)in cerebral cortex of the mice in various groups;ELISA method was used to determine the levels of Th17-and Treg-related cytokines including interleukin(IL)-17,IL-22,and IL-10 in serum of the mice;flow cytometry was used to determine the percentages of Th17 and Treg cells in peripheral blood of the mice in various groups,and the Th17/Treg ratio was calculated.Results:The serum of the mice induced with the GluN1 356-385 antigen peptide was positive for NMDAR IgG antibodies,indicating that the models were successfully established.Compared with control group,the neurological impairment score of the mice in model group was significantly increased(P<0.05),and the EB level in brain tissue was significantly increased(P<0.05);the fluorescence staining intensities of ZO-1 and Occludin in the cerebral cortex were decreased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins in the cerebral cortex were significantly decreased(P<0.05);the serum levels of IL-17 and IL-22 were significantly increased(P<0.05),while the IL-10 level was significantly decreased(P<0.05);the percentage of Th17 cells in peripheral blood was significantly increased(P<0.05),while the percentage of Treg cells was significantly decreased(P<0.05),and the Th17/Treg ratio was significantly increased(P<0.05).Compared with model group,the neurological impairment scores of the mice in XBJ-L and XBJ-H groups were significantly decreased(P<0.05),the EB levels in brain tissue were significantly decreased(P<0.05),the fluorescence staining intensities of ZO-1 and Occludin in cerebral cortex were increased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins were significantly increased(P<0.05);the levels of IL-17 and IL-22 in serum were significantly decreased(P<0.05),and the level of IL-10 was significantly increased(P<0.05);the percentages of Th17 cells in peripheral blood were significantly decreased(P<0.05),the percentages of Treg cells were significantly increased(P<0.05),and the Th17/Treg ratios were significantly decreased(P<0.05).Compared with XBJ-L group,the neurological function injury score of the mice in XBJ-H group was significantly decreased(P<0.05),the EB level in brain tissue was significantly decreased(P<0.05);the fluorescence staining intensities of ZO-1 and Occludin in the cerebral cortex were increased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins were significantly increased(P<0.05);the serum levels of IL-17 and IL-22 were significantly decreased(P<0.05),and the level of IL-10 was significantly increased(P<0.05);the percentage of Th17 cells in peripheral blood was significantly decreased(P<0.05),the percentage of Treg cells was significantly increased(P<0.05),and the Th17/Treg ratio was significantly decreased(P<0.05).Conclusion:Xuebijing injection can improve BBB injury,regulate Th17/Treg balance,and thereby alleviate the neurological functional damage in anti-NMDAR encephalitis.
4.Mechanism study on circASAP1 promoting proliferation and inhibiting apoptosis of osteosarcoma cells by targeting miR-4500
Yuehua GENG ; Lijun TANG ; Lusheng WEN ; Qiuyan LIN ; Jialun XU ; Limin YANG
Journal of Clinical Medicine in Practice 2025;29(19):58-64
Objective To investigate the regulatory effect of circular RNA ASAP1(circASAP1)targeting microRNA-4500(miR-4500)on the proliferation and apoptosis of osteosarcoma cells.Methods Human osteosarcoma cells(Saos-2)were selected as the experimental subjects and cul-tured in DMEM medium containing 10%fetal bovine serum while maintaining 5%CO2.The expres-sions of circASAP1 and miR-4500 in osteosarcoma tissues were detected using RT-qPCR.Saos-2 cells were transfected with si-NC(40 nmol/L),si-circASAP1(40 nmol/L),pcDNA(0.4 μg),pcDNA-circASAP1(0.4 μg),miR-NC(40 nmol/L),miR-4500 mimics(40 nmol/L),si-circASAP1+anti-miR-NC(40 nmol/L),and si-circASAP1+anti-miR-4500(40 nmol/L),respectively.Cell proliferation and apoptosis were assessed using CCK-8,colony formation assays,and flow cytometry.The interaction between circASAP1 and miR-4500 was analyzed using a dual-luciferase reporter assay.Results Compared with adjacent non-tumor tissues,the expression of circASAP1 was significantly upregulated,while that of miR-4500 was significantly downregulated in osteosarcoma tissues(P<0.001).Compared with the si-NC group,the si-circASAP1 group exhibited significantly decreased circASAP1 expression,optical density(OD)values,and the number of cell colonies(P<0.001).Compared with the si-NC group,the si-circASAP1 group showed significantly upregulated levels of cleaved-caspase3 protein,cleaved-caspase9 protein,and the apoptosis rate(P<0.001).StarBase search revealed specific binding sequences between miR-4500 and circASAP1.Co-transfection of miR-4500 mimics and WT-circASAP1 significantly reduced the relative luciferase activity of the cells[(0.34±0.03)versus(0.95±0.06),t=27.280,P<0.001].The expression of miR-4500 in the pcDNA-circASAP1 group was significantly lower than that in the pcDNA group,whereas the ex-pression of miR-4500 in the si-circASAP1 group was significantly higher than that in the si-NC group(P<0.001).Compared with the miR-NC group,the miR-4500 group exhibited significantly in-creased miR-4500 expression,cleaved-caspase3 protein levels,cleaved-caspase9 protein levels,and the apoptosis rate,while the OD values and the number of colonies significantly decreased(P<0.001).Compared with the si-circASAP1+anti-miR-NC group,the si-circASAP1+anti-miR-4500 group showed significantly decreased miR-4500 expression,cleaved-caspase3 protein levels,cleaved-caspase9 protein levels,and the apoptosis rate,while the OD values and the number of col-onies significantly increased(P<0.001).Conclusion The expression of circASAP1 is increased,while that of miR-4500 is decreased in osteosarcoma tissues.Moreover,circASAP1 promotes the proliferation and inhibits the apoptosis of osteosarcoma cells by targeting miR-4500.
5.Effects of Electroacupuncture on Skeletal Muscle Mitochondrial Dynamics in SAMP8 Mice
Ruolan WEN ; Wanqing GUO ; Weiguo DONG ; Limin CHEN ; Jinquan LI ; Chenguang WEN ; Lan LIN ; Feng WANG
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(2):104-109
Objective To observe the effects of electroacupuncture on the motor function and mitochondrial dynamics of skeletal muscle of SAMP8 mice;To explore the mechanism of electroacupuncture in improving the motor dysfunction of Alzheimer disease(AD)from the perspective of mitochondrial dynamics.Methods Totally 18 SAMP8 mice were divided into model group and electroacupuncture group,with 9 mice in each group,and the SAMR1 mice with the same age were set as control group."Baihui","Dazhui"and"Shenshu"were selected in the electroacupuncture group,and electroacupuncture was performed daily for 20 min,8 d as a course of treatment.Each course of treatment was separated by 2 d,for a total of 3 courses of treatment.The model group and the control group were not intervened.The motor function of mice was tested by grip strength test,suspension test,hind limb extension test and Morris water maze experiment.The morphology and structure of gastrocnemius were observed by HE staining,ATP content in gastrocnemius was determined by colorimetry,the mRNA expression of optic atrophy 1(OPA1),mitofusin 2(MFN2)and dynamin-related protein 1(DRP1)in gastrocnemius were detected by real-time quantitative PCR,the expressions of OPA1,MFN2 and DRP1 in gastrocnemius were detected by Western blot.Results Compared with the control group,the grip strength,the score in suspension test,and the average speed and maximum speed of Morris water maze experiment of mice in model group significantly decreased(P<0.01);the arrangement of fibers in the gastrocnemius muscle tissue was disordered,the gaps become wider,and the distribution of nuclei was uneven;the ATP content in the gastrocnemius muscle tissue was significantly decreased(P<0.01),the mRNA and protein expressions of OPA1 and MFN2 were significantly decreased(P<0.01),and the expression of DRP1 mRNA and protein significantly increased(P<0.01).Compared with the model group,the grip strength,the score in suspension test,and the average speed and maximum speed of Morris water maze experiment in electroacupuncture group significantly increased(P<0.01);the arrangement of gastrocnemius muscle tissue was relatively neat,the gaps become narrower,and the distribution of nuclei was more uniform;the ATP content in gastrocnemius muscle tissue significantly increased(P<0.01),while the mRNA and protein expressions of OPA1 and MFN2 significantly increased(P<0.05,P<0.01),the expression of DRP1 mRNA and protein significantly decreased(P<0.01).Conclusion Electroacupuncture can improve the skeletal muscle morphological structure and motor dysfunction of SAMP8 mice,and the mechanism may be related to the correction of skeletal muscle mitochondrial dynamic imbalance and the increase of skeletal muscle ATP content.
6.Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults (version 2024)
Qingde WANG ; Yuan HE ; Bohua CHEN ; Tongwei CHU ; Jinpeng DU ; Jian DONG ; Haoyu FENG ; Shunwu FAN ; Shiqing FENG ; Yanzheng GAO ; Zhong GUAN ; Hua GUO ; Yong HAI ; Lijun HE ; Dianming JIANG ; Jianyuan JIANG ; Bin LIN ; Bin LIU ; Baoge LIU ; Chunde LI ; Fang LI ; Feng LI ; Guohua LYU ; Li LI ; Qi LIAO ; Weishi LI ; Xiaoguang LIU ; Hongjian LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Yong QIU ; Limin RONG ; Yong SHEN ; Huiyong SHEN ; Jun SHU ; Yueming SONG ; Tiansheng SUN ; Yan WANG ; Zhe WANG ; Zheng WANG ; Hong XIA ; Guoyong YIN ; Jinglong YAN ; Wen YUAN ; Zhaoming YE ; Jie ZHAO ; Jianguo ZHANG ; Yue ZHU ; Yingjie ZHOU ; Zhongmin ZHANG ; Wei MEI ; Dingjun HAO ; Baorong HE
Chinese Journal of Trauma 2024;40(2):97-106
Ankylosing spondylitis (AS) combined with lower cervical fracture is often categorized into unstable fracture, with a high incidence of neurological injury and a high rate of disability and morbidity. As factors such as shoulder occlusion may affect the accuracy of X-ray imaging diagnosis, it is often easily misdiagnosed at the primary diagnosis. Non-operative treatment has complications such as bone nonunion and the possibility of secondary neurological damage, while the timing, access and choice of surgical treatment are still controversial. Currently, there are no clinical practice guidelines for the treatment of AS combined with lower cervical fracture with or without dislocation. To this end, the Spinal Trauma Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults ( version 2024) in accordance with the principles of evidence-based medicine, scientificity and practicality, in which 11 recommendations were put forward in terms of the diagnosis, imaging evaluation, typing and treatment, etc, to provide guidance for the diagnosis and treatment of AS combined with lower cervical fracture.
7.Efficacy and safety of eltrombopag in the treatment of primary immune thrombocytopenia: real-world data from a single medical center
Xifeng DONG ; Yalan LI ; Nianbin LI ; Weinan LIN ; Ting WANG ; Huaquan WANG ; Lijuan LI ; Wen QU ; Limin XING ; Hong LIU ; Yuhong WU ; Guojin WANG ; Jia SONG ; Jing GUAN ; Xiaoming WANG ; Zonghong SHAO ; Rong FU
Chinese Journal of Hematology 2024;45(3):271-276
Objective:This study aimed at investigating the efficacy and safety of eltrombopag in the treatment of adult primary immune thrombocytopenia (ITP) and evaluated the factors influencing its efficacy and side effects.Methods:A total of 198 patients with adult ITP who were admitted to Tianjin Medical University General Hospital between January 2018 and March 2022 were retrospectively analyzed. The efficacy of each starting dose of eltrombopag was evaluated, and adverse events were analyzed. The factors influencing efficacy were investigated, including sex, age, adult ITP type, platelet antibodies, and combined drug treatments.Results:Of the 198 patients, 70 males and 128 females with a median age of 45 years (18-88 years) were included; 130 (65.7%) had newly diagnosed adult ITP, 25 (12.6%) had persistent adult ITP, and 43 (21.7%) had chronic adult ITP. The bleeding event scores at baseline were assessed; 84.3% had scores of<4 and 15.7% had scores of ≥4. The eltrombopag response rate (initial response) at 6 weeks was 78.8% (complete response [CR]: 49.0%; CR1: 14.6%; CR2: 15.2%). The median response time to eltrombopag was 7 (7, 14) days. The initial response rates to 25, 50, and 75 mg eltrombopag were 74.1%, 85.9%, and 60.0%, respectively ( P=0.031). The initial response rate to the 50 mg dose was significantly higher than that of the 25-mg and 75-mg doses. Two patients received 100 mg as the starting dose, and their initial response was 0. Regarding dose adjustment, 70.7% of the patients remained on the starting dose, 8.6% underwent dose adjustment to 50 mg, and 6.1% underwent dose adjustment to 75 mg. Another two patients underwent dose adjustment to 100 mg. After dose adjustment, the persistent response rates were 83.6%, 85.3%, and 85.7% for the 25-, 50-, and 75-mg doses, respectively, with no significant difference. After dose adjustment, the sustained efficacy rate for the 100-mg dose (4 patients) was 100.0%. After 6 weeks of treatment with eltrombopag, the overall bleeding score of patients with ITP decreased. The number of patients with a score of ≥4 decreased to 0, the number of patients with a score of<4 decreased, and there was no significant change in the number of patients with a score of 1-2. The most common adverse event associated with eltrombopag was impaired liver function (7.7%). No thrombosis events or other adverse events were observed. ITP type and number of megakaryocytes significantly affected the initial response to eltrombopag. The initial response rates to eltrombopag for newly diagnosed adult ITP, persistent adult ITP, and chronic adult ITP were 85.3%, 56.0%, and 76.2%, respectively ( P=0.003). For megakaryocytes, the initial response rates were 61.8%, 87.1%, and 84.3% ( P=0.009) for the decreased, normal, and increased megakaryocyte groups, respectively. Conclusion:Eltrombopag, as a second-line or higher treatment for adult ITP, has a rapid onset of action and good safety. The initial response rate is significantly higher with a dose of 50 mg than with a dose of 25 mg. Patients with newly diagnosed ITP and those with normal or increased megakaryocyte numbers have a higher initial response rate to eltrombopag.
8.Analysis on the use of HIV post-exposure prophylaxis and related factors in men who have sex with men in Qingdao
Rong SU ; Limin ZHU ; Guihua HUANG ; Peilong LI ; Lin GE ; Meizhen LIAO ; Yong FU ; Xin SONG ; Dongmin LI
Chinese Journal of Epidemiology 2024;45(1):134-138
Objective:To understand the use of post-exposure prophylaxis (PEP) and analyze related factors among men who have sex with men (MSM) in Qingdao, and provide a reference for the AIDS prevention and control in this population.Methods:A cross-sectional survey conducted from April 2022 to February 2023. Relying on MSM social groups in Qingdao, a snowball sampling method was applied to recruit research subjects who met the inclusion criteria of age ≥18 years old, having had homosexual anal or oral sex in the past six months, and HIV-negative or infection status unknown. The sample size was estimated at 566. Data on demographic characteristics, sexual behavior characteristics, PEP use, and others of the research subjects was collected through on-site questionnaires. The logistic regression model was used to analyze the factors associated with using PEP.Results:A total of 811 participants were recruited, mainly aged 25-34 (53.6%, 435/811), unmarried (74.7%, 606/811), with an average monthly income of ≥5 000 yuan (52.2%, 423/811), and having lived in Qingdao for ≥10 years (75.6%, 613/811). The proportion of those who knew the HIV status of their sexual partners in the last six months was 67.1% (544/811), and those with HIV-positive partners were 3.6% (29/811). In the last six months, the proportion of participants who had group sex (86.4%, 701/811), unprotected anal sex (98.2%, 796/811), and use of club drugs (80.3%, 651/811) was high. Moreover, 28.4% (230/811) had used PEP. The multivariate logistic regression analysis showed that the factors related to the use of PEP included divorced or widowed (a OR=5.46,95% CI:1.96-15.17), average monthly income ≥5 000 yuan (a OR=2.04,95% CI:1.44-2.89), same-sex sexual orientation (a OR=0.40,95% CI:0.22-0.71), having HIV-positive sexual partners in the last six months (a OR=2.54,95% CI:1.13-5.71) and having been tested for HIV ≥3 times in the last six months (a OR=1.46,95% CI:1.04-2.06). Conclusions:The prevalence of risk behaviors among MSM in Qingdao was high, and the use of PEP was low. In the future, it is essential to increase HIV/AIDS prevention education among MSM, promote MSM to know the HIV status of their sexual partners, and reduce the prevalence of risk behaviors among this population. Additionally, explore medical insurance reimbursement plans for PEP to reduce utilization costs and promote the use of PEP by MSM after HIV exposure occurs as soon as possible.
9.Improvement effect of Xuebijing on brain tissue injury and Th17/Treg immune imbalance in cerebrospinal fluid in NMDA receptor encephalitis model mice
Lin CHEN ; Limin YAN ; Huaijie XING ; Min CHEN ; Xiaoyan LI ; Chaosheng ZENG
Journal of Jilin University(Medicine Edition) 2024;50(3):697-707
Objective:To discuss the effect of Xuebijing on brain tissue damage and immune imbalance of helper T lymphocyte 17(Th17)/regulatory T lymphocyte(Treg)in cerebrospinal fluid(CSF)of the N-methyl-D-aspartate(NMDA)receptor encephalitis model mice,and to clarify its therapeutic effect.Methods:Sixty healthy male C57BL/6J mice were randomly divided into control group,model group,low dose of Xuebijing group,and high dose of Xuebijing group,and there were 15 mice in each group.Except for control group,the mice in the other three groups were injected with the antigen combined with immunostimulation to establish the NMDA receptor encephalitis models.The mice in low and high doses of Xuebijing groups were injected intraperitoneally with 5 and 10 mL·kg-1 of Xuebijing injection,respectively.HE staining was used to observe the pathomorphology of brain tissue of the mice in various groups;TUNEL assay was used to detect the apoptotic rates of the neurons in hippocampus CA1 region of brain tissue of the mice in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of interleukin(IL)-6,IL-10,IL-17,and transforming growth factor β(TGF-β)in serum of the mice in various groups;flow cytometry was used to detect the percentages of Th17 and Treg cells in CSF of the mice in various groups;Western blotting method was used to detect the expression levels of retinoic acid-related orphan receptor γt(RORγt),forkhead box protein 3(Foxp3),IL-10,and IL-17 proteins in brain tissue of the mice in various groups;immunohistochemistry method was used to detect the rates of IL-17 and Foxp3 positive cells in brain tissue of the mice in various groups.Results:The HE staining results showed that the hippocampus CA1 region of brain tissue of the mice in control group had a clear structure without obvious lesions;compared with control group,the mice in model group showed partial pyramidal cell shrinkage,elongation of apical dendrites,loss of a few neurons,and sparse tissue in the hippocampus CA1 region of brain tissue;compared with model group,the mice in low and high doses of Xuebijing groups showed that the damage of the cells in the hippocampus CA1 region of brain tissue was decreased,and the morphological recovery,more orderly arrangement,and more significant improvement could be seen in hippocampus CA1 region of the mice in high dose of Xuebijing group.The TUNEL assay results showed that compared with control group,the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in model group was significantly increased(P<0.05);compared with model group,the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased(P<0.05);compared with low dose of Xuebijing group,the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in high dose of Xuebijing group was significantly decreased(P<0.05).The ELISA results showed that compared with control group,the levels of IL-6 and IL-17 in serum of the mice in model group were significantly increased(P<0.05),while the levels of IL-10 and TGF-β were significantly decreased(P<0.05);compared with model group,the levels of IL-6 and IL-17 in serum of the mice in low and high doses of Xuebijing groups were significantly decreased(P<0.05),while the levels of IL-10 and TGF-β were significantly increased(P<0.05);compared with low dose of Xuebijing group,the levels of IL-6 and IL-17 in serum of the mice in high dose of Xuebijing group were significantly decreased(P<0.05),while the levels of IL-10 and TGF-β were significantly increased(P<0.05).The flow cytometry results showed that compared with control group,the percentage of CD4+IL-17A+Th17 cells in CSF of the mice in model group was significantly increased(P<0.05),while the percentage of CD25+Foxp3+Treg cells was significantly decreased(P<0.05);compared with model group,the percentages of CD4+IL-17A+Th17 cells in CSF of the mice in low and high doses of Xuebijing groups were significantly decreased(P<0.05),while the percentage of CD25+Foxp3+Treg cells was significantly increased(P<0.05);compared with low dose of Xuebijing group,the percentage of CD4+IL-17A+Th17 cells in CSF of the mice in high dose of Xuebijing group was significantly decreased(P<0.05),while the percentage of CD25+Foxp3+Treg cells was significantly increased(P<0.05).The Western blotting results showed that compared with control group,the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in model group were significantly increased(P<0.05),while the expression levels of Foxp3 and IL-10 proteins were significantly decreased(P<0.05);compared with model group,the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased(P<0.05),while the expression levels of Foxp3 and IL-10 proteins were significantly increased(P<0.05);compared with low dose of Xuebijing group,the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in high dose of Xuebijing group were significantly decreased(P<0.05),while the expression levels of Foxp3 and IL-10 proteins were significantly increased(P<0.05).The immunohistochemistry results showed that compared with control group,the rate of IL-17 positive cells in brain tissue of the mice in model group was significantly increased(P<0.05),while the rate of Foxp3 positive cells was significantly decreased(P<0.05);compared with model group,the rates of IL-17 positive cells in brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased(P<0.05),while the rates of Foxp3 positive cells were significantly increased(P<0.05);compared with low dose of Xuebijing group,the rate of IL-17 positive cells in brain tissue of the mice in high dose of Xuebijing group was significantly decreased(P<0.05),while the rate of Foxp3 positive cells was significantly increased(P<0.05).Conclusion:Xuebijing can effectively ameliorate the brain tissue injury,regulate the cytokine levels,and intervene in immune imbalance of Th17/Treg in the mice with anti-NMDA receptor encephalitis.
10.Development and reliability and validity test of the Pediatric Nursing Care Deficiency Scale
Chunmin LIN ; Ru YIN ; Limin PANG
Chinese Journal of Modern Nursing 2024;30(30):4171-4175
Objective:To develop a Pediatric Nursing Care Deficiency Scale (PNCDS) and evaluate its reliability and validity.Methods:Based on theories of nursing care deficiency and considering pediatric nursing characteristics, the scale was developed following standard procedures and principles. This included semi-structured interviews, expert evaluations, a pilot survey, and a large-scale survey to finalize the scale.Results:The final version of the PNCDS comprised four dimensions with 25 items. The content validity index (CVI) of the overall scale was 0.937, with item-level CVIs ranging from 0.857 to 1.000. Exploratory factor analysis revealed that the factor loadings for all items were greater than 0.5, and the four factors accounted for a cumulative variance of 66.456%. Confirmatory factor analysis indicated a good model fit for the final scale. The overall Cronbach's α coefficient for the scale was 0.917, while the Cronbach's α coefficients for the four dimensions were 0.861, 0.923, 0.881, and 0.909, respectively.Conclusions:The PNCDS demonstrated good reliability and validity and provides a useful tool for assessing pediatric nursing care deficiencies and aiding in the development of intervention strategies.

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