Pancreatic ductal adenocarcinoma(PDAC)is one of the most aggressive solid malignancies.A specific mechanism of its metastasis has not been established.In this study,we investigated whether Neural Wiskott-Aldrich syndr...Pancreatic ductal adenocarcinoma(PDAC)is one of the most aggressive solid malignancies.A specific mechanism of its metastasis has not been established.In this study,we investigated whether Neural Wiskott-Aldrich syndrome protein(N-WASP)plays a role in distant metastasis of PDAC.We found that N-WASP is markedly expressed in clinical patients with PDAC.Clinical analysis showed a notably more distant metastatic pattern in the N-WASP-high group compared to the N-WASP-low group.N-WASP was noted to be a novel mediator of epithelialmesenchymal transition(EMT)via gene expression profile studies.Knockdown of N-WASP in pancreatic cancer cells significantly inhibited cell invasion,migration,and EMT.We also observed positive association of lysyl oxidase-like 2(LOXL2)and focal adhesion kinase(FAK)with the N-WASP-mediated response,wherein EMT and invadopodia function were modulated.Both N-WASP and LOXL2 depletion significantly reduced the incidence of liver and lung metastatic lesions in orthotopic mouse models of pancreatic cancer.These results elucidate a novel role for N-WASP signaling associated with LOXL2 in EMT and invadopodia function,with respect to regulation of intercellular communication in tumor cells for promoting pancreatic cancer metastasis.These findings may aid in the development of therapeutic strategies against pancreatic cancer.展开更多
Background： Cryopyrin-associated periodic syndrome （CAPS） is a group of rare, heterogeneous autoinflammatory disease characterized by interleukin （IL）-1β-mediated systemic inflammation and clinical symptoms invo...Background： Cryopyrin-associated periodic syndrome （CAPS） is a group of rare, heterogeneous autoinflammatory disease characterized by interleukin （IL）-1β-mediated systemic inflammation and clinical symptoms involving skin, joints, central nervous system, and eyes. It encompasses a spectrum of three clinically overlapping autoinflammatory syndromes including familial cold autoinflammatory syndrome, Muckle-Wells syndrome （MWS）, and neonatal-onset multisystem inflammatory disease. CAPS is associated with gain-of-function missense mutations in NOD-like receptor family pyrin domain-containing protein 3 （NLRP3）, the gene encoding NLRP3. Moreover, most mutations leading to MWS occurred in exon 3 ofNLRP3 gene. Here, we reported a novel mutation occurred in exon 1 ofNLRP3 gene in an MWS patient and attempted to explore the pathogenic mechanism. Methods： Genetic sequence analysis of NLRP3 was performed in an MWS patient who presented with periodic lever, arthralgia, and multiform skin lesions. NLRP3 was also analyzed in this patient＇s parents and 50 healthy individuals. Clinical examinations including X-ray examination, skin biopsy, bone marrow aspiration smear, and blood test of C-reactive protein （CRP）, erythrocyte sedimentation rate （ESR）, serum levels oflL-1β, immunoglobulin E （lgE）, antineutrophil cytoplasmic antibodies, antinuclear antibodies, and extractable nuclear antigen were also analyzed. The protein structure of mutant NLRP3 inflammasome was calculated by SWISS-MODEL software. Proteins of wild type and mutant components ofNLRP3 inflammasome were expressed and purified, and the interaction abilities between these proteins were tested by surface plasmon resonance （SPR） assay. Results： X-ray examination showed no abnormality in the patient＇s knees. Laboratory tests indicated an elevation of CRP （233.24 nag/L） and ESR （67 mm/h） when the patient had fever. Serum IL-1β increased to 24.37 pg/ml, and serum lgE was higher than 2500.00 IU/ml. Other blood tests were normal. Bone marrow aspiration smear was normal. A novel point mutation c.92A〉T in exon 1 of NLRP3 gene was identified, which caused a p.D31V mutation in pyrin domain （PYD） of NLRP3. SPR assay showed that this point mutation may strengthen the interaction between the PYD of NLRP3 and the PYD of the apoptosis-associated speck-like protein. The mutation c.92A〉T in exon 1 of the NLRP3 gene was not lbund in the patient＇s parents and 50 healthy individuals. Conclusions： The rnutation c.92A〉T in exon 1 of the NLRP3 gene is a novel mutation associated with MWS. The p.D31V mutation might promote the activation ofNLRP3 inflammasome and induce MWS in this patient.展开更多
目的:探讨急性冠脉综合征(acute coronary syndrome,ACS)患者血清补体C1肿瘤坏死因子相关蛋白家族12(complement C1 tumor necrosis factor-associated protein family 12,CTRP12)水平与影响ACS及冠脉病变严重程度因素的关系。方法:选择...目的:探讨急性冠脉综合征(acute coronary syndrome,ACS)患者血清补体C1肿瘤坏死因子相关蛋白家族12(complement C1 tumor necrosis factor-associated protein family 12,CTRP12)水平与影响ACS及冠脉病变严重程度因素的关系。方法:选择2021年11月—2022年8月于江苏省丹阳市人民医院心血管内科行冠状动脉造影的患者共116例,其中ACS 81例,非ACS 35例。收集血液标本检测血清CTRP12及其他血液生化指标。心脏超声测量心外膜脂肪厚度(epicardial fat thickness,EFT)。比较两组血清CTRP12水平和EFT,分析CTRP12与其他因素的相关性。采用SYNTAX(Synergy between PCI with TAXUSTM and Cardiac Surgery)评分评估冠脉病变严重程度,将其分为SYNTAX评分≤22分和SYNTAX评分>22分,Logistic回归分析影响ACS及冠脉病变严重程度的因素。结果:ACS组患者男性比例、吸烟比例、EFT、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、WBC计数、中性粒细胞计数、单核细胞计数均高于非ACS组(均P<0.05),血清CTRP12水平低于非ACS组(P<0.05),EFT较非ACS组增加(P<0.05)。ACS组患者中SYNTAX评分≤22分者血清CTRP12水平高于SYNTAX评分>22分者(P<0.05),EFT小于SYNTAX评分>22分者(P<0.05)。相关性分析提示CTRP12与EFT呈负相关(r=-0.557,P<0.05)。Logistic回归提示CTRP12和EFT是ACS的独立影响因素(β=-2.107、1.607,均P<0.05)。CTRP12和EFT是ACS患者冠脉病变严重程度的影响因素(β=-1.085、1.740,均P<0.05),在校正年龄、性别、BMI、吸烟、糖尿病、高血压、LDL-C后,CTRP12和EFT仍是ACS患者冠脉病变严重程度的影响因素(β=-1.033、2.109,均P<0.05)。结论:ACS患者血清CTRP12水平显著降低,且与EFT呈负相关。EFT是ACS及冠脉病变严重程度的独立危险因素,而CTRP12为ACS的保护因素。展开更多
OBJECTIVES: To study the expression of inflammatory signal in local prostate tissue of chronic pelvic pain syndrome(CPPS) rats by electroacupuncture(EA) of Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6...OBJECTIVES: To study the expression of inflammatory signal in local prostate tissue of chronic pelvic pain syndrome(CPPS) rats by electroacupuncture(EA) of Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6), and to explore the possible mechanism of anti-inflammatory and analgesic effects of EA. METHODS : A total of 36 Sprague-Dawley male rats were randomly divided into three groups: control, model and EA(n=12 rats/group). The CPPS model was made by injection of CFA into ventral lobes of the prostate(0.1 m L). Electric acupuncture apparatus was applied to stimulate Guanyuan(CV4), Zhongji(CV3), bilateral Huiyang(BL35) and Sanyinjiao(SP6) acupoints in EA group. The general condition of rats was observed and the prostate index(PI) was calculated. The thermal pain threshold was collected after each therapeutic course. Histopathological changes of the prostate tissue were examined by hematoxylin-eosin staining method. The expression levels of tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β) and prostaglandin E2(PGE2) in prostatic homogenates were measured by enzyme linked immunosorbent assay(ELISA). Moreover, the expression levels of purinergic 2X7 receptor(P2X7R), NOD-like receptor pyrin domain-containing 3(NLRP3), caspase-1 and interleukin-18(IL-18) m RNA were quantified by quantitative real-time polymerase chain reaction. RESULTS: Compared with control group, the PI of rats increased, and the thermal pain threshold decreased significantly in model group. The morphological structure of prostate tissues of rats in model group was severely damaged with a large number of inflammatory cells infiltration. Additionally, the levels of TNF-α, IL-1β and PGE2 were higher, and the expressions of P2X7R, NLRP3, caspase-1 and IL-18 m RNA were higher than those in control group. After EA treatment, the PI was significantly decreased, the thermal pain threshold was significantly increased, and the tissue damage was significantly improved. The expressions of inflammatory cytokines were lower in EA group, and expression of P2X7R/NLRP3 pathway was down-regulated. CONCLUSION: The effect of EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can improve inflammation and pain symptoms of CPPS rats induced by Complete Freund’s adjuvant(CFA). This suggests that EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can produce antiinflammatory analgesia effect by preventing the activation of P2X7R/NLRP3 signal pathway, inhibit the release of inflammatory cytokines in CPPS rats, which may provide a putative novel target for the treatment of CPPS.展开更多
目的探讨子痫前期产妇胎盘组织存在的氧化应激对胎盘滋养细胞Wiskott-Aldrich综合征关联蛋白2（Wiskott-Aldrich syndrome related protein2，WAVE2）表达的影响。方法研究对象为2011年8月15日至2012年2月23日在重庆医科大学附属第一医...目的探讨子痫前期产妇胎盘组织存在的氧化应激对胎盘滋养细胞Wiskott-Aldrich综合征关联蛋白2（Wiskott-Aldrich syndrome related protein2，WAVE2）表达的影响。方法研究对象为2011年8月15日至2012年2月23日在重庆医科大学附属第一医院分娩的子痫前期产妇20例及正常足月产妇23例（对照组）。剖宫产术后取胎盘组织，采用免疫组织化学法检测胎盘组织WAVE2的定位及分布。采用蛋白质印迹法检测胎盘组织WAVE2的表达，实时荧光定量聚合酶链反应法检测胎盘中WAVE2mRNA的表达，采用组织匀浆法检测2组产妇胎盘组织活性氧（reactive oxygen species，ROs）水平，并对胎盘组织中ROS水平与wAVE2表达水平进行相关性分析。（2）通过构建体外滋养细胞缺氧复氧损伤模型，模拟缺血再灌注诱导的氧化应激损伤。人妊娠早期绒毛外滋养细胞株HTR-8／SVneo细胞贴壁完全后分别置于常氧（常氧组）和缺氧复氧条件（缺氧复氧组）下48h，采用流式细胞仪分析细胞内ROS水平。采用Transwell实验观察细胞迁移侵袭状况。采用细胞免疫荧光法和蛋白质印迹法检测HTR-8／SVneo细胞内WAVE2的定位及表达水平的变化。采用独立样本t检验及Pearson相关检验对数据进行分析。结果（1）子痫前期组24h尿蛋白、母体收缩压及舒张压均高于对照组，新生儿出生体重及胎盘重量均低于对照组[24h尿蛋白：（1．96±0．24）g与（0．08士0．05）g，t=19．436；母体收缩压：（154±13）mmHg与（98±11）FilmHg，t=11．324；母体舒张压：（105±14）mmHg与（69±8）mmHg，t=9．612；新生儿出生体重：（2446±187）g与（3207±233）g，t=16．307；胎盘重量：（432±53）g与（536±67）g，t=14．562；均P〈O．05]。子痫前期组WAVE2mRNA表达低于对照组E（0．28±0．07）与（1．01±0．02），t=12．747]，WAVE2相对表达量低于对照组[（0．63±0．08）与（1．34±0．19），t=11．648]，ROS水平高于对照组[（144．22±12．32）nmol／（mg·prot）与（75．17±8．71）nmol／（mg·prot），t=20．467]，差异均有统计学意义（均P〈0．05）。子痫前期组胎盘组织中ROS水平与wAVE2表达水平呈显著负相关（r=-0．726，P=0．000）。（2）在常氧组中，HTR-8／svneo细胞内的ROS水平为（82．9±5．8）％，而缺氧复氧组为（155．6±8．1）％，高于常氧组（t=12．747，P〈0．05）。缺氧复氧48h后的HTR=8／SVneo细胞与常氧组相比，迁移和侵袭力明显降低[缺氧复氧组分别为（58．4±4．2）％和（51．9±3．3）％，常氧组为100％，t值分别为11．034和13．839，P均〈O．os]。细胞免疫荧光检测结果显示，WAVE2定位于滋养细胞的细胞质中。缺氧复氧48h后，HTR=8／SVneo细胞中WAVE2的表达强度明显弱于常氧组（O．37±0．05与0．76±0．06，t=8．631，P〈0．05）。结论子痫前期胎盘组织中存在过度氧化应激，与胎盘组织中WAVE2表达下调密切相关。缺氧复氧致氧化应激损伤可下调滋养细胞WAVE2的表达；WAVE2表达下调可能损伤滋养细胞迁移侵袭能力，从而参与子痫前期的发生发展。展开更多
基金supported by a National Research Foundation of Korea(NRF)grant funded by the Korean Government,Ministry of Science and ICT(MSIT)(2016R1C1B102207,2022R1A2C1004141 and 2022R1A2C-1091712)the National R&D Program for Cancer Control through the National Cancer Center(NCC)funded by the Ministry of Health&Welfare,Republic of Korea(HA22C0053000022).
文摘Pancreatic ductal adenocarcinoma(PDAC)is one of the most aggressive solid malignancies.A specific mechanism of its metastasis has not been established.In this study,we investigated whether Neural Wiskott-Aldrich syndrome protein(N-WASP)plays a role in distant metastasis of PDAC.We found that N-WASP is markedly expressed in clinical patients with PDAC.Clinical analysis showed a notably more distant metastatic pattern in the N-WASP-high group compared to the N-WASP-low group.N-WASP was noted to be a novel mediator of epithelialmesenchymal transition(EMT)via gene expression profile studies.Knockdown of N-WASP in pancreatic cancer cells significantly inhibited cell invasion,migration,and EMT.We also observed positive association of lysyl oxidase-like 2(LOXL2)and focal adhesion kinase(FAK)with the N-WASP-mediated response,wherein EMT and invadopodia function were modulated.Both N-WASP and LOXL2 depletion significantly reduced the incidence of liver and lung metastatic lesions in orthotopic mouse models of pancreatic cancer.These results elucidate a novel role for N-WASP signaling associated with LOXL2 in EMT and invadopodia function,with respect to regulation of intercellular communication in tumor cells for promoting pancreatic cancer metastasis.These findings may aid in the development of therapeutic strategies against pancreatic cancer.
基金This work was supported by the grant from the National Natural Science Foundation of China （No. 81201267）.
文摘Background： Cryopyrin-associated periodic syndrome （CAPS） is a group of rare, heterogeneous autoinflammatory disease characterized by interleukin （IL）-1β-mediated systemic inflammation and clinical symptoms involving skin, joints, central nervous system, and eyes. It encompasses a spectrum of three clinically overlapping autoinflammatory syndromes including familial cold autoinflammatory syndrome, Muckle-Wells syndrome （MWS）, and neonatal-onset multisystem inflammatory disease. CAPS is associated with gain-of-function missense mutations in NOD-like receptor family pyrin domain-containing protein 3 （NLRP3）, the gene encoding NLRP3. Moreover, most mutations leading to MWS occurred in exon 3 ofNLRP3 gene. Here, we reported a novel mutation occurred in exon 1 ofNLRP3 gene in an MWS patient and attempted to explore the pathogenic mechanism. Methods： Genetic sequence analysis of NLRP3 was performed in an MWS patient who presented with periodic lever, arthralgia, and multiform skin lesions. NLRP3 was also analyzed in this patient＇s parents and 50 healthy individuals. Clinical examinations including X-ray examination, skin biopsy, bone marrow aspiration smear, and blood test of C-reactive protein （CRP）, erythrocyte sedimentation rate （ESR）, serum levels oflL-1β, immunoglobulin E （lgE）, antineutrophil cytoplasmic antibodies, antinuclear antibodies, and extractable nuclear antigen were also analyzed. The protein structure of mutant NLRP3 inflammasome was calculated by SWISS-MODEL software. Proteins of wild type and mutant components ofNLRP3 inflammasome were expressed and purified, and the interaction abilities between these proteins were tested by surface plasmon resonance （SPR） assay. Results： X-ray examination showed no abnormality in the patient＇s knees. Laboratory tests indicated an elevation of CRP （233.24 nag/L） and ESR （67 mm/h） when the patient had fever. Serum IL-1β increased to 24.37 pg/ml, and serum lgE was higher than 2500.00 IU/ml. Other blood tests were normal. Bone marrow aspiration smear was normal. A novel point mutation c.92A〉T in exon 1 of NLRP3 gene was identified, which caused a p.D31V mutation in pyrin domain （PYD） of NLRP3. SPR assay showed that this point mutation may strengthen the interaction between the PYD of NLRP3 and the PYD of the apoptosis-associated speck-like protein. The mutation c.92A〉T in exon 1 of the NLRP3 gene was not lbund in the patient＇s parents and 50 healthy individuals. Conclusions： The rnutation c.92A〉T in exon 1 of the NLRP3 gene is a novel mutation associated with MWS. The p.D31V mutation might promote the activation ofNLRP3 inflammasome and induce MWS in this patient.
文摘目的:探讨急性冠脉综合征(acute coronary syndrome,ACS)患者血清补体C1肿瘤坏死因子相关蛋白家族12(complement C1 tumor necrosis factor-associated protein family 12,CTRP12)水平与影响ACS及冠脉病变严重程度因素的关系。方法:选择2021年11月—2022年8月于江苏省丹阳市人民医院心血管内科行冠状动脉造影的患者共116例,其中ACS 81例,非ACS 35例。收集血液标本检测血清CTRP12及其他血液生化指标。心脏超声测量心外膜脂肪厚度(epicardial fat thickness,EFT)。比较两组血清CTRP12水平和EFT,分析CTRP12与其他因素的相关性。采用SYNTAX(Synergy between PCI with TAXUSTM and Cardiac Surgery)评分评估冠脉病变严重程度,将其分为SYNTAX评分≤22分和SYNTAX评分>22分,Logistic回归分析影响ACS及冠脉病变严重程度的因素。结果:ACS组患者男性比例、吸烟比例、EFT、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、WBC计数、中性粒细胞计数、单核细胞计数均高于非ACS组(均P<0.05),血清CTRP12水平低于非ACS组(P<0.05),EFT较非ACS组增加(P<0.05)。ACS组患者中SYNTAX评分≤22分者血清CTRP12水平高于SYNTAX评分>22分者(P<0.05),EFT小于SYNTAX评分>22分者(P<0.05)。相关性分析提示CTRP12与EFT呈负相关(r=-0.557,P<0.05)。Logistic回归提示CTRP12和EFT是ACS的独立影响因素(β=-2.107、1.607,均P<0.05)。CTRP12和EFT是ACS患者冠脉病变严重程度的影响因素(β=-1.085、1.740,均P<0.05),在校正年龄、性别、BMI、吸烟、糖尿病、高血压、LDL-C后,CTRP12和EFT仍是ACS患者冠脉病变严重程度的影响因素(β=-1.033、2.109,均P<0.05)。结论:ACS患者血清CTRP12水平显著降低,且与EFT呈负相关。EFT是ACS及冠脉病变严重程度的独立危险因素,而CTRP12为ACS的保护因素。
基金Supported by Fundamental Research Funds for the Central Universities Project:RNA Sequencing Technology Screening the Mechanism of Acupuncture on Pain in Chronic Pelvic Pain Syndrome Rats (2020-JYB-XJSJJ-018)。
文摘OBJECTIVES: To study the expression of inflammatory signal in local prostate tissue of chronic pelvic pain syndrome(CPPS) rats by electroacupuncture(EA) of Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6), and to explore the possible mechanism of anti-inflammatory and analgesic effects of EA. METHODS : A total of 36 Sprague-Dawley male rats were randomly divided into three groups: control, model and EA(n=12 rats/group). The CPPS model was made by injection of CFA into ventral lobes of the prostate(0.1 m L). Electric acupuncture apparatus was applied to stimulate Guanyuan(CV4), Zhongji(CV3), bilateral Huiyang(BL35) and Sanyinjiao(SP6) acupoints in EA group. The general condition of rats was observed and the prostate index(PI) was calculated. The thermal pain threshold was collected after each therapeutic course. Histopathological changes of the prostate tissue were examined by hematoxylin-eosin staining method. The expression levels of tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β) and prostaglandin E2(PGE2) in prostatic homogenates were measured by enzyme linked immunosorbent assay(ELISA). Moreover, the expression levels of purinergic 2X7 receptor(P2X7R), NOD-like receptor pyrin domain-containing 3(NLRP3), caspase-1 and interleukin-18(IL-18) m RNA were quantified by quantitative real-time polymerase chain reaction. RESULTS: Compared with control group, the PI of rats increased, and the thermal pain threshold decreased significantly in model group. The morphological structure of prostate tissues of rats in model group was severely damaged with a large number of inflammatory cells infiltration. Additionally, the levels of TNF-α, IL-1β and PGE2 were higher, and the expressions of P2X7R, NLRP3, caspase-1 and IL-18 m RNA were higher than those in control group. After EA treatment, the PI was significantly decreased, the thermal pain threshold was significantly increased, and the tissue damage was significantly improved. The expressions of inflammatory cytokines were lower in EA group, and expression of P2X7R/NLRP3 pathway was down-regulated. CONCLUSION: The effect of EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can improve inflammation and pain symptoms of CPPS rats induced by Complete Freund’s adjuvant(CFA). This suggests that EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can produce antiinflammatory analgesia effect by preventing the activation of P2X7R/NLRP3 signal pathway, inhibit the release of inflammatory cytokines in CPPS rats, which may provide a putative novel target for the treatment of CPPS.
