饲喂不同浓度黄曲霉毒素B_1饲料对异育银鲫成鱼的生长和毒素积累的影响

以含不同浓度黄曲霉毒素B1(AFB1)的配合饲料饲喂异育银鲫(Carassius auratus gibelio)成鱼56d,研究异育银鲫成鱼[(122.3±0.7)g]生长、生理反应、肝脏组织学变化、卵巢发育以及鱼体各组织中的AFB1的毒素积累状况。实验分为5个实验组,不同实验组饲料中AFB1含量分别为0、5、20、50、500μg/kg饲料(实测值分别为2.59、4.12、12.39、46.23、454.07μg/kg饲料),每个处理3个平行。在整个实验过程中各实验组均未表现出外部形态和行为异常,各组存活率均达到100%。各实验组异育银鲫成鱼终末体重、摄食率(FR)、特定生长率(SGR)和饲料效率(FE)均无显著差异。饲料AFB1水平对异育银鲫血清总胆固醇(TC)含量、血清谷丙转氨酶(GPT)、谷草转氨酶(GOT)和碱性磷酸酶(AKP)活性均无显著影响。各毒素组血清超氧化物岐化酶(SOD)活性与对照无显著差异。各毒素组肝脏和卵巢均未见明显的组织学病理变化。肌肉和性腺中的AFB1积累量低于FDA食品安全限定标准(5μg/kg)。肝胰脏中的AFB1积累和饲料中的AFB1水平呈对数关系。饲喂AFB1≥50μg/kg饲料使异育银鲫成鱼肝脏AFB1积累超过安全限量标准。结果表明,异育银鲫成鱼至少可耐受AFB1含量达500μg/kg饲料(实测值:454.07μg/kg饲料)56d

间充质细胞外泌体促进小鼠胰岛内皮细胞血管生成的研究

目的探讨间充质细胞（MSC）外泌体对低氧条件下胰岛内皮细胞（MS-1）血管生成的影响。方法 MSC无血清低氧条件培养48 h,超滤离心法富集条件培养基中的外泌体,采用电镜和Western Blot的方法进行鉴定;通过血管形成试验比较分析不同条件下:常氧培养组（NOR组,21%O2、5%CO2）、低浓度氧培养组（HYP组,2%O2、5%CO2）、外泌体+低浓度氧共培养组（HYP+EXO组,2%O2、5%CO2）,MS-1细胞的血管形成能力;image J软件分析血管形成长度;PCR、Q-PCR检测血管内皮生长因子（VEGF） RNA水平的表达,Western Blot检测VEGF、HIF1α蛋白水平表达以及mTOR信号通路激活情况。采用单因素方差分析和SNK-q检验统计学分析。结果超滤离心法富集的MSC条件培养基中的外泌体,大小为30～100 nm,表达CD9,CD63,CD81等外泌体表面标志物;血管形成试验结果显示,低氧促进MS-1血管生成,HYP+EXO组形成明显的血管网状结构;HYP+EXO组血管形成相对长度（2386.0±137.7）像素与NOR组（393.3±174.2）像素和HYP组（1467.0±230.0）像素相比增强,差异有统计学意义（t=12.30,P=0.0065;t=15.74,P=0.0040）; PCR结果显示,HYP+EXO组VEGF相对表达量（20.26±9.972）较常氧对照组（1.000）和低氧组（6.521±3.501）均增强,差异有统计学意义（t=5.462,P=0.0009;t=4.238,P=0.0038）;同时,Western Blot结果显示VEGF蛋白水平表达升高,HIF1-α表达上调,mTOR发生磷酸化。结论 MSC外泌体可促进低氧条件下的小鼠胰岛内皮细胞血管生成。MSC外泌体可能通过上调HIF1-α,调节VEGF表达,激活mTOR信号通路,促进胰岛内皮细胞血管生成。国家自然科学基金青年项目(81601618);;福建省自然科学基金面上项目(2016J01582、2016J01580、2018J01349);;福建省科技创新联合资金重大项目(2017Y9127

Cloning and expression analysis of lipopolysaccharide-induced TNF-α(LITAF) of Japanese scallop (Mizuhopecten yessoensis)

脂多糖诱导的肿瘤坏死因子(lIPOPOlySACCHArIdE-InduCEd Tnf-AlPHA fACTOr,lITAf)是一类重要的炎症细胞因子,在先天性免疫系统中发挥重要的介质作用。文章根据虾夷扇贝lITAf基因EST序列,应用rACE技术克隆了虾夷扇贝lITAf全长CdnA,对序列及编码的氨基酸进行生物信息学分析。结果显示,该基因CdnA全长1 551 bP,其5′非编码区包含76 bP,3′非编码区包含1 001 bP;开放阅读框(Orf)为474 bP,编码157个氨基酸,氨基酸序列中存在一个保守的lITAf结构域;理论分子量16.99 kdA,等电点为6.24。lITAf基因序列为3 698 bP,由3个外显子和两个内含子组成。利用实时荧光定量PCr技术分析lITAf在虾夷扇贝不同组织、不同胚胎发育阶段以及鳗弧菌(VIbrIO AnguIllAruM)刺激后各时间段的表达情况。结果表明:lITAf基因在所检测的6个成体组织中均有表达,其中肾脏的表达量最高;胚胎发育的7个时期中,担轮幼体时期表达量最高;菌刺激36 H实验组与对照组的表达量差异大。lITAf基因是lITAf家族的一员,推测lITAf基因参与虾夷扇贝的先天性免疫反应。The lipopolysaccharide-induced TNF-alpha factor(LITAF) is an inflammatory cytokine,which plays an important role in innate immunity system.Based on the expressed sequence tag(EST) of Japanese scallop(Mizuhopecten yessoensis),the cDNA of LITAF gene was amplified using rapid amplification of cDNA ends(RACE) approach.Results showed that the full-length cDNA of LITAF is 1 551 bp consisting of a 5′ untranslated region(UTR) of 76 bp,a 3' UTR of 1 001 bp,and an open reading frame(ORF) of 474 bp encoding a polypeptide of 157 amino acids,and there is a conserved LITAF domain in amino acid sequences.The estimated molecular mass is 16.99 kDa and the theoretical isoelectric point is 6.24.The total length of LITAF is 3 698 bp,which includes three exons and two introns.Real-time quantitative PCR was carried out to measure LITAF mRNA expression in adult tissues and monitor mRNA expression patterns during embryonic development after bacteria(Vibrio anguillarum) challenged.The expression level of LITAF mRNA was detected in all the adult tissues with the highest in the kidneys.The trochophore owns the highest expression level of LITAF in embryonic development.LITAF expression showed significant difference(P<0.01)between the control and bacteria challenged specimens at 36 h.These results suggest that the LITAF should be a member of the LITAF family that perhaps involved in the innate immune response of Japanese scallop.国家海洋公益性行业科研专项(编号:200805037);国家自然科学基金项目(编号:31140073)资

骨髓间充质细胞联合PDMS支架构建移植胰岛微环境的实验研究

目的为了提高移植胰岛的活性和功能,构建适合移植胰岛生存的微环境。方法采用聚二甲基硅氧烷(PDMS)和氯化钠晶体构建三维支架,联合骨髓间充质细胞(MSCs)、纤维蛋白和胰岛共同构建迷你\"人工胰腺\"。采用链脲佐菌素(STZ)诱导的糖尿病大鼠移植模型评价效果,将\"人工胰腺\"移植到糖尿病大鼠大网膜内,对照组行假手术,术后隔天监测移植大鼠血糖水平;数据采用t检验和曼-惠特尼U检验。结果用PDMS构建的三维巨孔支架,支架内可见大量不规则孔洞空间。胰岛和MSCs可成功装载入支架内,HE染色结果显示,支架孔内存在胰岛,胰岛周围包绕有MSCs。糖尿病大鼠大网膜内移植结果显示,移植后各时间点(1,3,5,7 d),\"人工胰腺\"移植组糖尿病大鼠血糖水平分别为(278.70±86.06) mg/dl、(323.50±44.29) mg/dl、(283.30±74.00) mg/dl、(304.80±13.33) mg/dl,较假手术对照组(606.00±52.40) mg/dl、(589.70±55.78) mg/dl、(615.00±54.84) mg/dl、(630.30±48.17) mg/dl均降低,差异具有统计学意义(t=7.96、9.15、8.82,U=0.00,P均<0.01)。结论 MSCs联合PDMS三维支架构建的微环境,可为移植胰岛提供生存的环境,为临床开展胰岛移植提供新的策略

Video-assisted Thoracoscopic 3D Mode Operation for Solitary Pulmonary Nodules

目的探讨三维胸腔镜手术(3d VIdEO-ASSISTEd THOrACIC SurgEry,3d-VATS治疗孤立性肺结节(SOlITAry PulMOnAry nOdulE,SPn)的效果。方法回顾性分析2013年3月~2014年3月50例SPn的资料,采用3d-VATS手术模式楔形切除结节,根据快速病理结果决定是否行肺叶切除加淋巴结清扫术。统计手术时间(去除快速冰冻时间)、术后24 H引流量、总引流量、引流管拔除时间、淋巴结清扫数及术后并发症等。结果 3d-VATS模式下,50例均行肺结节楔形切除,其中23例病理为恶性,继续行肺癌根治术,手术顺利。肺癌根治术手术时间(62±12)MIn,术中出血量(35±5)Ml,清扫淋巴结(19±3)个,术后24 H引流量(120±20)Ml,术后胸管引流时间(4±1)d,术后住院时间(7±2)d。并发症3例,其中术后肺炎2例,阵发性心房纤颤1例,均治愈。无围手术期死亡。随访2~12个月,平均6.3月。1例术后3个月脑转移,1例术后5个月肺癌复发。结论胸腔镜3d模式下治疗SPn是一种新的选择方式,安全可行,值得推广应用。Objective To evaluate the effects of surgical treatment for solitary pulmonary nodules under thoracoscopic 3D mode(3D-VATS).Methods A total of 50 cases of solitary pulmonary nodules from March 2013 to March 2014 were retrospectively analyzed.Intraoperative wedge pulmonary resection with 3D-VATS was utilized.According to intraoperative pathological findings,lobectomy plus lymph node dissection was given or not.Intraoperative time( minus fast freezing time),drainage volume for 24 h,total drainage volume,drainage tube removal time,number of lymph node dissected,and postoperative complications were recorded.Results Under 3D-VATS mode,50 cases of solitary pulmonary nodules were treated with wedge resection,including 23 cases of malignant pathology receiving radical resection, which was smoothly.The radical resection time( lung lobectomy plus lymphadenectomy) was(62 ± 12) min,the bleeding volume was(35 ± 5) ml,the lymphadenectomy number was 19 ± 3,the drainage volume for 24 h was(120 ± 20) ml,the postoperative chest tube removal time was(4 ± 1) days,and the postoperative hospital stay was(7 ± 2) days.Postoperative complications occurred in 3 cases,including 2 cases of pneumonia and 1 case of paroxysmal atrial fibrillation.No perioperative deaths were observed.All the cases were followed up for 2- 12 months,with an average of 6.3 months.Brain metastases was found in 1 case at the third postoperative month and recurrence of lung cancer was noted in 1 case at the fifth postoperative month.Conclusion Thoracoscopic 3D mode treatment for solitary pulmonary nodules is a new,safe,and feasible alternative and should be widely applied

Combined Use of Thoracoscopy and Laparoscopy in Total Laryngectomy for Cervical Esophageal Carcinoma

目的探讨胸、腹腔镜联合全喉切除治疗颈段食管的可行性和疗效。方法 2009年1月~2014年7月胸、腹腔镜联合全喉切除治疗33例颈段食管癌。胸腔镜下分离食管、腹腔镜下管胃成形、全喉切除、气管永久造口、胃咽吻合术。结果胸部手术时间40~66 MIn,平均53 MIn;腹部手术时间35~51 MIn,平均44 MIn;颈部手术时间128~150 MIn,平均139 MIn。术中出血量130~270 Ml,平均150 Ml。术后住院时间8~14 d,平均12 d。病理均为鳞状细胞癌,其中高分化2例,中分化19例,中-低分化7例,低分化5例。切缘病理学检查无癌组织残留。31例淋巴结转移。并发症:吻合口漏2例,喉返神经损伤3例,肺部感染6例,胃排空障碍2例,吻合口狭窄1例,无死亡病例。33例随访1个月~5年,术后1、3、5年生存率分别为87.9%、54.5%、45.5%。结论颈段食管癌应采取积极的手术治疗,胃咽吻合术是颈段食管癌切除后较为理想的修复手段。Objective To investigate clinical feasibility and efficacy of combined use of thoracoscopy and laparoscopy in total laryngectomy for cervical esophageal carcinoma.Methods Clinical data of 33 patients with cervical esophageal carcinoma undergoing surgical treatment in our department from January 2009 to July 2014 were analyzed retrospectively.The esophagus was separated under thoracoscopy.And laparoscopic gastroplasty,total laryngectomy,tracheal permanent colostomy,and gastric pharyngeal anastomosis were performed.Results The thoracoscopic operation time was 40- 66 min( mean,53 min),the laparoscopic operation time was 35- 51 min( mean,44 min),and the cervical operation time was 128- 150 min( mean,139 min).The blood loss was 130- 270 ml( mean,150 ml).The postoperative hospital stay was 8- 14 d( mean,12 d).Pathological examinations showed squamous cell carcinoma in all the cases,including 2 cases of highly differentiated carcinoma,19 cases of moderately differentiated carcinoma,7 cases of moderately or lowly differentiated carcinoma,and 5 cases of lowly differentiated carcinoma.No residual cancer was found at cutting edges pathologically.Among the 33 cases,lymph node metastasis was found in 31 cases.Complications included 2 cases of anastomotic fistula,3 cases of recurrent laryngeal nerve injury,6 cases of pulmonary infection,2cases of delayed gastric emptying,and 1 case of anastomotic stenosis.There was no death.All the patients were followed up for 1months to 5 years.The survival rates at 1,3,and 5 postoperative year were 87.9%,54.5%,and 45.5%,respectively.Conclusions Cervical esophageal carcinoma should be surgically treated actively.Gastric pharyngeal anastomosis is an ideal option for the repair of cervical esophageal cancer resection

Trisomy 21-induced Dysregulation of Microglial Homeostasis in Alzheimer’s Brains is Mediated by USP25

阿尔茨海默病（Alzheimer’s disease, AD）是一种最为常见的与记忆、认知能力退化相关的渐进性神经退行性疾病。唐氏综合征（Down’s syndrome, DS）是早发型阿尔茨海默病的一个重要风险因素，作为最常见的智力障碍遗传疾病，厦门大学医学院神经科学研究所王鑫教授团队揭示了治疗阿尔茨海默病和唐氏综合征新的治疗靶点，并且在小鼠模型上利用USP25小分子抑制剂成功地改善了阿尔茨海默病小鼠的认知功能，缓解了神经退行性病变的病理进程。该研究工作由王鑫教授指导完成，厦门大学医学院助理教授郑秋阳和博士生李桂林完成主要实验工作，王世华、朱琳、高月、邓青芳、张洪峰、张丽珊、吴美玲、狄安洁参与了部分研究工作。厦门大学医学院许华曦、赵颖俊和孙灏教授在研究过程中给予大力帮助和支持，清华大学董晨教授提供了Usp25基因敲除小鼠，厦门大学附属妇女儿童医院周裕林教授和郑良楷博士帮助收集了脑组织样品。Down syndrome (DS), caused by trisomy of chromosome 21, is the most significant risk factor for early-onset Alzheimer’s disease (AD); however, underlying mechanisms linking DS and AD remain unclear. Here, we show that triplication of homologous chromosome 21 genes aggravates neuroinflammation in combined murine DS-AD models. Overexpression of USP25, a deubiquitinating enzyme encoded by chromosome 21, results in microglial activation and induces synaptic and cognitive deficits, whereas genetic ablation of Usp25 reduces neuroinflammation and rescues synaptic and cognitive function in 5×FAD mice. Mechanistically, USP25 deficiency attenuates microglia-mediated proinflammatory cytokine overproduction and synapse elimination. Inhibition of USP25 reestablishes homeostatic microglial signatures and restores synaptic and cognitive function in 5×FAD mice. In summary, we demonstrate an unprecedented role for trisomy 21 and pathogenic effects associated with microgliosis as a result of the increased USP25 dosage, implicating USP25 as a therapeutic target for neuroinflammation in DS and AD.This work was supported by the National Natural Science Foundation of China (31871077, 81822014, and 81571176 to X.W.; 81701130 to Q.Z.), the National Key R&D Program of China (2016YFC1305900 to X.W.), the Natural Science Foundation of Fujian Province of China (2017J06021 to X.W.), the Fundamental Research Funds for the Chinese Central Universities (20720150061 to X.W.), and the BrightFocus Foundation (A2018214F to Yingjun Zhao). 该研究工作得到国家重点研发计划项目、国家自然科学基金、福建省自然科学基金、厦门大学校长基金的资助和支持