立体定向大鼠脑内SHG-44人脑胶质瘤模型的建立

目的：探讨建立大鼠SHG-44脑胶质瘤模型的方法。方法：选用雄性Wistar大鼠,接种前连续3 d用地塞米松1 mg/100 g体重灌胃;在立体定向条件下选取大鼠脑右侧尾状核区为靶点,接种2×10⁵个处于对数生长期的SHG-44细胞,接种后观察大鼠生长状态,分别于第1、2周进行核磁共振(MRI)检查。在实验第2周时解剖标本,行组织病理和GFAP免疫组化检查。 结果：接种1周后核磁共振检查,脑内形成实体瘤;HE染色组织病理证实是胶质瘤,GFAP免疫组化阳性;成瘤率约60%。 结论：用预先免疫抑制的方法,成功建立了大鼠脑内人胶质瘤模型。     

Heterologous expression of the invertebrate FMRFamide-gated sodium channel as a mechanism to selectively activate mammalian neurons

Considerable effort has been directed toward the development of methods to selectively activate specific subtypes of neurons. Focus has been placed on the heterologous expression of proteins that are capable of exciting neurons in which they are expressed. Here we describe the heterologous expression of the invertebrate FMRFamide (H-phenylalanine-methionine-arginine-phenylalanine-NH(2)) -gated sodium channel from Helix aspersa (HaFaNaC) in hippocampal slice cultures. HaFaNaC was co-expressed with a fluorescent protein (green fluorescent protein (GFP), red fluorescent protein from Discosoma sp (dsRed) or mutated form of red fluorescent protein from Discosoma sp (tdTomato)) in CA3 pyramidal neurons of rat hippocampal slice cultures using single cell electroporation. Pressure application of the agonist FMRFamide to HaFaNaC-expressing neuronal somata produced large prolonged depolarizations and bursts of action potentials (APs). FMRFamide responses were inhibited by amiloride (100 muM). In contrast, pressure application of FMRFamide to the axons of neurons expressing HaFaNaC produced no response. Fusion of GFP to the N-terminus of HaFaNaC showed that GFP-HaFaNaC was absent from axons. Bath application of FMRFamide produced persistent AP firing in HaFaNaC-expressing neurons. This FMRFamide-induced increase in the frequency of APs was dose-dependent. The concentrations of FMRFamide required to activate HaFaNaC-expressing neurons were below that required to activate the homologous acid sensing ion channel normally found in mammalian neurons. Furthermore, the mammalian neuropeptides neuropeptide FF and RFamide-related peptide-1, which have amidated RF C-termini, did not affect HaFaNaC-expressing neurons. Antagonists of NPFF receptors (BIBP3226) also had no effect on HaFaNaC. Therefore, we suggest that heterologous-expression of HaFaNaC in mammalian neurons could be a useful method to selectively and persistently excite specific subtypes of neurons in intact nervous tissue.     

人脐血源性内皮祖细胞血管发生活性及其参与人恶性胶质瘤细胞异种移植瘤血管新生的作用

目的 观察人脐血源性内皮祖细胞(EPC)血管发生能力和在恶性胶质瘤血管新生过程中的作用.方法 应用密度梯度离心法分离新鲜人脐血的单个核细胞,接种于EGM-2培养液中培养获得EPC.取生长到第7～10天的细胞进行CD34和VEGFR-2免疫荧光双标染色.检测血管内皮生长因子(VEGF)刺激下EPC增殖活性、迁移能力和体外形成小管样结构的能力.采用人恶性胶质瘤细胞系U87在免疫缺陷小鼠进行皮下移植,于接种肿瘤后第7天经尾静脉注射EPC(每只5×103),并于接种肿瘤后第28天取材检测肿瘤微血管和EPC组织分布及定位,采用抗人CD31和抗鼠CD31免疫荧光双标记肿瘤微血管,计算人源性EPC来源的血管占肿瘤血管网的比例.结果 培养的细胞在第7～10天时可见条索样结构,生长并逐渐融合形成铺路石样排列的单层细胞,表达内皮细胞标记物CD34和VEGFR-2.在VEGF刺激下EPC具有较强的增殖活性、迁移能力和体外形成小管样结构的能力.外源性EPC能特异性归巢到异种移植瘤组织并形成新生血管,占肿瘤血管网的(18.68±1.32)%.结论 EPC在体内外具有形成血管能力,并参与异种移植瘤血管新生,提示其在恶性肿瘤血管新生过程中具有重要作用,并可能参与肿瘤微血管构筑表型异质性.     

血红素加氧酶-1基因转染对非协调性异种心脏移植物的影响

目的 观察血红素加氧酶-1(HO-1)基因转染对非协调性异种心脏移植物的影响,并探讨其可能的保护机制.方法 采用颈部袖套血管吻合法联合应用中华眼镜蛇毒因子(CVF)建立稳定的豚鼠-SD大鼠非协调性异种异位心脏移植模型.供体豚鼠和受体SD大鼠各24只,每组各8只随机配对分为Ⅰ组(对照组)、Ⅱ组(空白载体组)和Ⅲ组(HO-...     

接受供精人工授精夫妇生育压力和生活质量研究

目的：调查接受供精人工授精（AID）夫妇的生育压力与生活质量，了解生育压力和生活质量的关系，为这些夫妇进行心理和生活质量干预时提供理论依据。方法：采用患者夫妇一般情况调查表、生育压力量表（FPI量表）和生活质量量表（FertiQoL量表）对北京大学第三医院2018年5—9月生殖中心门诊就诊的129对准备接受AID助孕的夫妇进行横断面调查，比较夫妇双方生育压力和生活质量的差异，分析生育压力和生活质量之间的关系。结果：接受AID的女性患者FPI总分为（159.00±17.52）分，FertiQoL总分为（62.27±9.26）分；男性配偶FPI总得分为（153.91±19.96）分，男性配偶FertiQoL总分为（65.12±11.62）分。女性FPI总分高于男性（t=2.436，P=0.016），女性无子女压力维度得分高于男性（t=3.145，P=0.002）。男性FertiQol总分高于女性（t=-2.881，P=0.005），女性情感反应维度得分高于男性（t=3.681，P=0.000），男性身心关系维度得分高于女性（t=-11.401，P=0.000）。Pearson 相关分析结果显示，接受AID助孕的女性FPI总分及性压力、夫妻关系、父母角色需求压力三个维度与其生活质量呈负相关（P＜0.05），接受AID男性配偶FPI总分和社会压力、性压力、夫妻关系维度、父母角色需求压力这4个维度与其生活质量呈负相关（P＜0.05）。多重线性回归显示，生育压力、居住地、文化程度是影响接受AID助孕女性患者生活质量的重要影响因素（F=3.485，R2=0.168，P＜0.05），生育压力、文化程度、家庭月收入是影响接受AID助孕男性配偶生活质量的重要影响因素（F=4.390，R2=0.203，P＜0.01）。结论：接受AID女性患者生育压力高于男性配偶，生活质量低于男性配偶；生育压力是影响AID夫妇生活质量的重要因素，夫妇双方生育压力越大，生活质量越低。     

Heterologous fibrin sealant potentiates axonal regeneration after peripheral nerve injury with reduction in the number of suture points

The use of suture associated with heterologous fibrin sealant has been highlighted for reconstruction after peripheral nerve injury, having the advantage of being safe for clinical use. In this study we compared the use of this sealant associated with reduced number of stitches with conventional suture after ischiatic nerve injury. 36 Wistar rats were divided into 4 groups: Control (C), Denervated (D), ischiatic nerve neurotmesis (6 mm gap); Suture (S), epineural anastomosis after 7 days from neurotmesis, Suture + Fibrin Sealant (SFS), anastomosis with only one suture point associated with Fibrin Sealant. Catwalk, electromyography, ischiatic and tibial nerve, soleus muscle morphological and morphometric analyses were performed. The amplitude and latency values of the Suture and Suture + Fibrin Sealant groups were similar and indicative of nerve regeneration.The ischiatic nerve morphometric analysis in the Suture + Fibrin Sealant showed superior values related to axons and nerve fibers area and diameter when compared to Suture group. In the Suture and Suture + Fibrin Sealant groups, there was an increase in muscle weight and in fast fibers frequency, it was a decrease in the percentage of collagen compared to group Denervated and in the neuromuscular junctions, the synaptic boutons were reestablished.The results suggest a protective effect at the lesion site caused by the fibrin sealant use. The stitches reduction minimizes the trauma caused by the needle and it accelerates the surgical practice. So the heterologous fibrin sealant use in nerve reconstruction should be considered.     

Retrospective study of malignant phyllodes tumors of the breast: Younger age,prior fibroadenoma surgery,malignant heterologous elements and surgical margins may predict recurrence

PurposeThe potential recurrence rate of malignant phyllodes tumors (MPTs) of the breast is high, and the prognostic factors are still unclear. We therefore aim to study the factors affecting the outcome of MPTs.MethodsA retrospective review of MPT patients treated from 2006 to 2020 at our institution was conducted. Univariate and multivariate Cox proportional hazard models were used to examine the influence of different variables on RFS. Moreover, significant prognostic factors were combined to construct the nomogram to predict the probability of relapse occurring in MPT patients. The 5-year and 10-year RFS rates were estimated using the Kaplan–Meier method.ResultsDuring the study period, 188 MPT patients were identified. The presence of malignant heterologous elements was observed in 23 (12.2%) patients with MPT, and the patients with malignant heterologous elements who received chemotherapy had longer RFS, which could reduce the risk of recurrence (p = 0.022). Recurrence occurred in 56/188 (29.8%) patients, of whom 47 experienced local recurrence and 11 experienced distant metastases. The 5-year and 10-year cumulative RFS rates were 77.5% and 70.1%, respectively. Age (p = 0.041), fibroadenoma surgery history (p = 0.004), surgical margins (p = 0.001) and malignant heterologous elements (p < 0.001) were independent risk factors for postoperative RFS. Subsequently, a nomogram was built, with a C-index of 0.64 (95% CI: 0.629–0.661), to predict the risk of recurrence.ConclusionThe results of this study showed that younger age, fibroadenoma surgery history, malignant heterologous elements and surgical margins <1 cm predict a higher incidence of recurrence in MPT patients. Patients with malignant heterologous elements treated with chemotherapy could have a reduced risk of recurrence.     

猪内源性逆转录病毒体外感染人胚肾细胞系HEK-293的初步研究

目的对猪内源性逆转录病毒(PERV)体外感染人胚肾细胞系HEK-293的能力进行检测。方法建立体外感染人胚肾细胞系HEK-293的方法,用免疫电镜的方法观察PERV粒子,聚合酶链反应(PCR)、逆转录聚合酶链反应(RT—PCR)检测与PERV共培养24h后HEK-293细胞基因组中PERV前病毒基因的整合、表达并对PERV亚型进行鉴定,激光共聚焦显微镜检测PERV—gag蛋白的表达并进行定量分析。结果电镜下可观察到PERV的圆形病毒粒子;与PERV共培养24h后HEK-293细胞基因组中检测到PERV前病毒DNA,且mRNA也有效表达,PERV亚型为PERV—A,B型;激光共聚焦显微镜观察到PERV—gag蛋白在感染细胞HEK-293的胞质表达,但表达量下降。结论PERV具有体外感染人胚肾细胞系HEK-293的能力,病毒蛋白可以有效表达,因此在猪到人的异种器官移植研究中对PERV可能引起的人兽共患病进行安全性评价是极有必要的。     

两种移植材料治疗肿瘤刮除后骨缺损的临床研究

目的 比较观察脱蛋白牛松质骨复合自体红骨髓和自体髂骨移植修复骨肿瘤或瘤样病变引起的腔洞性骨缺损的临床疗效。方法 按随机数字表分为实验组和对照组,实验组接受脱蛋白牛松质骨复合自体红骨髓移植治疗,对照组接受自体髂骨移植治疗。结果 实验组有６３例,对照组有６２例,接受了骨移植术治疗,经平均随访２０个月,两组腔洞性骨缺损的愈合率及各种并发症均无显著性差异（Ｐ〉０．０５）,但自体骨移植组手术时间明显长于牛松     

裸小鼠肺癌模型的建立

目的：建立皮下异种肿瘤移植的裸小鼠肺癌模型。方法：采用培养细胞移植法,将人肺腺癌细胞系Ａ５４９ 接种于ＢａｌＢ／ＣＡ ｎｕ 品系裸小鼠背部皮下,观察肿瘤移植成功率,测量瘤体大小,并进行病理形态学检查。结果：裸小鼠皮下异种肺癌移植成功率为６０％ ～８５％ 。结论