目的观察不同时间点神经痛小鼠海马CA3区外周单核细胞的浸润,阐述这一现象对小鼠神经痛及焦虑样行为的影响。方法采用健康雄性C57小鼠,将小鼠随机分为假手术组(sham)、坐骨神经分支选择性损伤(SNI)模型组(SNI)、CCR2抑制剂RS102895处理...目的观察不同时间点神经痛小鼠海马CA3区外周单核细胞的浸润,阐述这一现象对小鼠神经痛及焦虑样行为的影响。方法采用健康雄性C57小鼠,将小鼠随机分为假手术组(sham)、坐骨神经分支选择性损伤(SNI)模型组(SNI)、CCR2抑制剂RS102895处理组(SNI+RS102895)和小胶质细胞活化抑制剂米诺环素(MC)处理组(SNI+MC),共4组。其中假手术组和模型组均进一步分为7 d、14 d和18 d组,SNI+RS102895和SNI+MC组均在第18天时取材。手术诱发神经痛,采用机械缩足阈(PWTs)测定不同时点痛阈,所有小鼠处死前2 d进行高架十字迷宫(EPM)实验,处死前1 d进行旷场实验(OFT)观察焦虑样行为变化。免疫荧光法检测小鼠海马CA3脑区白细胞分化抗原45(CD45)的表达及与小胶质细胞标记物离子钙接头蛋白分子1(IBA-1)、跨膜蛋白119(TMEM119),星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)和神经元标记物神经元核抗原(NeuN)的共表达,流式细胞术检测14 d SNI小鼠全脑单核细胞的百分比。18 d小鼠SNI后第5~16天分别灌胃给予MC 90 mg/(kg·d)、RS1028955 mg/(kg·d)与生理盐水,观察阻断单核细胞浸润对小鼠神经痛、焦虑样行为及海马CA3区CD45及IBA-1的表达的影响。结果7 d和14 d组小鼠SNI后1 d PWTs下降,持续到处死前(P<0.01)。假手术组CD45表达极少;与同时段假手术组比较,7 d SNI小鼠CD45表达无增加((P>0.05),14 d SNI小鼠CD45表达显著上升(P<0.01),且只与IBA-1和TMEM119有少量共表达,与GFAP、NeuN无共表达。14 d SNI小鼠全脑单核细胞百分比显著上升(P<0.01)。抑制小胶质细胞激活与抑制CCR2表达均能减少SNI小鼠CA3区CD45的表达(P<0.01),且能提高SNI小鼠机械痛阈(P<0.01)并缓解焦虑样行为(P<0.01)。结论SNI诱发神经痛14 d后小鼠海马CA3区有外周单核细胞的浸润,该现象可能参与了神经痛的维持及促进焦虑样行为的发生。展开更多
Objective:Serological tests are widely used for scrub typhus diagnosis;however,their limitations are evident.This study aims to assess their practical value in clinical settings.Methods:We analyzed the data of adult p...Objective:Serological tests are widely used for scrub typhus diagnosis;however,their limitations are evident.This study aims to assess their practical value in clinical settings.Methods:We analyzed the data of adult patients with suspected scrub typhus who visited a tertiary care hospital in the Republic of Korea from September to December from 2019 to 2021.The included patients had an acute fever and at least one of the following ten secondary findings:myalgia,skin rash,eschar,headache,thrombocytopenia,increased liver enzyme levels,lymphadenopathy,hepatomegaly,splenomegaly,and pleural effusion.The diagnoses were grouped as scrub typhus or other diseases by two infectious disease physicians.Results:Among 136 patients who met the eligibility criteria,109 had scrub typhus and 27 had different diseases.Single and paired total antibodies using immunofluorescence assay(IFA),and total antibodies using immunochromatography-based rapid diagnostic testing(ICT)were measured in 98%,22%,and 75%of all patients,respectively.Confirmation using paired samples for scrub typhus was established at a median of 11[interquartile range(IQR)10-16]days following the first visit.Among the 82 admitted patients,the median admission time was 9(IQR 7-13)days.According to IFA,58(55%)patients with scrub typhus had total immunoglobulin titers≥1:320,while 23(85%)patients with other disease had titers<1:320.Positive ICT results were observed in 64(74%)patients with scrub typhus and 10(67%)patients with other diseases showed negative ICT results.Conclusions:Serological testing for scrub typhus is currently insufficient for decision-making in clinical practice.展开更多
文摘目的观察不同时间点神经痛小鼠海马CA3区外周单核细胞的浸润,阐述这一现象对小鼠神经痛及焦虑样行为的影响。方法采用健康雄性C57小鼠,将小鼠随机分为假手术组(sham)、坐骨神经分支选择性损伤(SNI)模型组(SNI)、CCR2抑制剂RS102895处理组(SNI+RS102895)和小胶质细胞活化抑制剂米诺环素(MC)处理组(SNI+MC),共4组。其中假手术组和模型组均进一步分为7 d、14 d和18 d组,SNI+RS102895和SNI+MC组均在第18天时取材。手术诱发神经痛,采用机械缩足阈(PWTs)测定不同时点痛阈,所有小鼠处死前2 d进行高架十字迷宫(EPM)实验,处死前1 d进行旷场实验(OFT)观察焦虑样行为变化。免疫荧光法检测小鼠海马CA3脑区白细胞分化抗原45(CD45)的表达及与小胶质细胞标记物离子钙接头蛋白分子1(IBA-1)、跨膜蛋白119(TMEM119),星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)和神经元标记物神经元核抗原(NeuN)的共表达,流式细胞术检测14 d SNI小鼠全脑单核细胞的百分比。18 d小鼠SNI后第5~16天分别灌胃给予MC 90 mg/(kg·d)、RS1028955 mg/(kg·d)与生理盐水,观察阻断单核细胞浸润对小鼠神经痛、焦虑样行为及海马CA3区CD45及IBA-1的表达的影响。结果7 d和14 d组小鼠SNI后1 d PWTs下降,持续到处死前(P<0.01)。假手术组CD45表达极少;与同时段假手术组比较,7 d SNI小鼠CD45表达无增加((P>0.05),14 d SNI小鼠CD45表达显著上升(P<0.01),且只与IBA-1和TMEM119有少量共表达,与GFAP、NeuN无共表达。14 d SNI小鼠全脑单核细胞百分比显著上升(P<0.01)。抑制小胶质细胞激活与抑制CCR2表达均能减少SNI小鼠CA3区CD45的表达(P<0.01),且能提高SNI小鼠机械痛阈(P<0.01)并缓解焦虑样行为(P<0.01)。结论SNI诱发神经痛14 d后小鼠海马CA3区有外周单核细胞的浸润,该现象可能参与了神经痛的维持及促进焦虑样行为的发生。
基金the Korea Health Technology R&D Project through the Korea Health Industry Development Institute(KHIDI),funded by the Ministry of Health&Welfare,Republic of Korea(grant no.HI22C0306).
文摘Objective:Serological tests are widely used for scrub typhus diagnosis;however,their limitations are evident.This study aims to assess their practical value in clinical settings.Methods:We analyzed the data of adult patients with suspected scrub typhus who visited a tertiary care hospital in the Republic of Korea from September to December from 2019 to 2021.The included patients had an acute fever and at least one of the following ten secondary findings:myalgia,skin rash,eschar,headache,thrombocytopenia,increased liver enzyme levels,lymphadenopathy,hepatomegaly,splenomegaly,and pleural effusion.The diagnoses were grouped as scrub typhus or other diseases by two infectious disease physicians.Results:Among 136 patients who met the eligibility criteria,109 had scrub typhus and 27 had different diseases.Single and paired total antibodies using immunofluorescence assay(IFA),and total antibodies using immunochromatography-based rapid diagnostic testing(ICT)were measured in 98%,22%,and 75%of all patients,respectively.Confirmation using paired samples for scrub typhus was established at a median of 11[interquartile range(IQR)10-16]days following the first visit.Among the 82 admitted patients,the median admission time was 9(IQR 7-13)days.According to IFA,58(55%)patients with scrub typhus had total immunoglobulin titers≥1:320,while 23(85%)patients with other disease had titers<1:320.Positive ICT results were observed in 64(74%)patients with scrub typhus and 10(67%)patients with other diseases showed negative ICT results.Conclusions:Serological testing for scrub typhus is currently insufficient for decision-making in clinical practice.
