Matrix Metalloproteinases in Breast Cancer

Abstract: Matrix metalloproteinases (MMPs) represent a class of enzymes able to degrade numerous extracellular matrix macromolecules facilitating tumor invasion and metastasis. The principal MMPs involved in breast pathology are analyzed with their various roles and functions: gelatinases A and B, stromelysin-3, collagenase-3, and MT-MMP1 (membrane type MMP). In vivo and in vitro studies clearly demonstrate an important cooperation between tumor and stromal cells for the expression of these MMPs in breast carcinomas. The large expression of MMPs plead in favor of a major role of these enzymes in breast carcinoma progression and their detection may be used in some cases as a prognostic indicator. Studies now are in progress, directed toward the modulation of these MMPs and their inhibitors with new therapeutic agents to block tumoral invasion and metastasis due to these enzymes.?     

Transplantation of cultured sympathetic ganglionic neurons into Parkinsonian rat brain: Survival and function of graft

Summary The superior cervical ganglia (SCG) of newborn rats, which had been cultured as expiants for varying periods of time, were transplanted into the striatum of rats with unilateral 6-hydroxydopamine lesions of the nigrostriatal dopamine pathway to examine the survival and functional properties of the sympathetic neurons maintained in long-term culture prior to grafting. In the rats given the SCG cultured in vitro for 2 weeks, apomorphine-induced rotational behaviour was satisfactory reduced. The rats receiving the SCG from 4-week-old cultures showed only modest behavioural changes. The grafting of the SCG cultured for 6 weeks in vitro did not affect the rotational behaviour. These behavioural data corresponded with the histological assessment of the graft survival by use of catecholamine histofluorescence. The present results suggest the critical time period in vitro which might allow the cultured sympathetic neurons to be successfully grafted.     

A neural network confirms that physical exercise reverses EEG changes in depressed rats

The use of an artificial neural network (ANN) system to differentiate the EEG power density spectra in depressed from normal rats was tried. The beneficial effects of chronic physical exercise in reducing the effects of stress and therefore depression was also to be tested in animals by the same method. In this study, rats were divided into 4 groups, subjected to (i) chronic stress (D group); (ii) chronic exercise by treadmill running (EO group); (iii) exercise with stress (ES group) and (iv) handling (C group). The prefrontal cortical EEG, EMG and EOG were recorded simultaneously on paper and the digitized EEG signals were also stored in the hard-disk of a PC-AT through an ADC. After filtering the digitize signals, the EEG power spectra were calculated by an FFT routine. Three successive 4 s artefact-free epochs were averaged. The REM and NREM sleep periods as well as the awake period signals were analyzed separately. The FFT values from each of the 3 states, in the 4 groups of animals were tested by an ANN with 30 first layer neurons and a 2nd layer of a majority-vote-taker. The ANN could distinguish the depressed from the normal rats' EEG very well in REM (99%) sleep, NREM (95%) sleep and awake (81%) states. In most of the cases it identified the exercised rats' EEG as normal.     

基于术前脾脏CT影像组学列线图预测胃癌的浆膜浸润

目的：联合临床检验指标及影像学特征构建一种能够术前识别胃癌浆膜浸润的模型。方法：选取2015年1月至2019年12月温州医科大学附属第一医院经病理证实的656例胃癌患者,采用随机数字表法分为建模组（394例）和验证组（262例）。收集建模组患者的脾脏影像学资料,对收集的数据进行套索回归并选取差异有统计学意义的特征来构建浆膜浸润预测模型。在最大约登指数下取肿瘤浸润风险评分截断值将患者分为高危组（238例）和低危组（418例）,然后与其他浸润相关因素如BMI、年龄、性别、高血压、糖尿病等进行单变量和多变量Logistic回归分析,结合显著的独立影响因素共同建立可视化的浆膜浸润预测列线图。结果：将患者以肿瘤浸润评分≤-0.335分为低危组,＞-0.335为高危组,经验证组验证,建模组和验证组的诊断准确性较为一致（P＜0.001）。经浸润影响因素的单变量和多变量Logistic回归分析发现,影像组学肿瘤浸润评分（OR=2.9,95%CI=2.1～4.2,P＜0.001）、术前低白蛋白（OR=1.3,95%CI=1.2～3.1,P=0.003）、血小板与淋巴细胞比值（OR=1.8,95%CI=1.2～2.7,P=0.004）、肿瘤分化程度（OR=2.6,95%CI= 1.8～3.7,P＜0.001）是浆膜浸润的独立影响因素。基于这4个指标建立的预测模型能够较为准确地预测浆膜浸润风险,其AUC值为0.733。结论：基于脾脏影像的肿瘤浸润评分联合其他临床因素可准确预测胃癌浆膜浸润与否,提高诊断精度。     

沉默circROBO1通过下调KRAS抑制鼻咽癌CNE2细胞侵袭与肺转移

目的探讨沉默环状RNA hsa_circ_0124696(circROBO1)对鼻咽癌CNE2细胞侵袭与肺转移的影响机制。 方法qRT-PCR检测鼻咽癌及癌旁组织中circROBO1表达。采用干扰小RNA(siRNA)沉默鼻咽癌CNE2细胞中circROBO1的表达,Transwell及HE染色检测circROBO1对CNE2细胞迁移能力和体内肺转移的影响。TargetScan在线软件预测circROBO1下游miR-217与下游靶基因KRAS的靶向结合位点,双荧光素酶报告基因实验验证两者之间的靶向调控关系。蛋白免疫印迹检测siRNA沉默CNE2细胞中circROBO1表达对KRAS的影响。 结果鼻咽癌组织中circROBO1表达高于癌旁组织(P＜0.05)。与转染si-circNC的对照组相比,si-circROBO1组鼻咽癌CNE2细胞侵袭与体内肺转移能力均显著降低(P＜0.05)。circROBO1下游miR-217与KRAS之间存在靶向结合位点,并且circROBO1可影响KRAS的蛋白和mRNA表达量。 结论沉默circROBO1通过miR-217下调KRAS抑制鼻咽癌CNE2细胞侵袭与肺转移。     

miR-21靶向E2F1对三阴性乳腺癌细胞恶性生物学活性及裸鼠肿瘤抑制率的影响

目的 探究miR-21靶向E2F1对三阴性乳腺癌细胞恶性生物学活性及裸鼠肿瘤抑制率的影响。方法将MDA-MB-231细胞分为5组,即MDA-MB-231组、miR-21 inhibitor组、miR-NC inhibitor组、siRNA-E2F1组和siRNA-NC组。检测细胞中miR-21表达（RT-PCR法）;分别检测细胞增殖（MTT法）、侵袭（Transwell法）、迁移（划痕实验）和凋亡能力（流式细胞仪）;检测细胞中E2F1蛋白表达;检测miR-21与E2F1的靶向关系（双荧光素酶实验报告）。结果MDA-MB-231细胞中miR-21表达明显高于MCF10A细胞（P<0.05）;miR-21 inhibitor组细胞细胞中miR-21表达明显低于MDA-MB-231组（P<0.05）。与MDA-MB-231组相比,miR-21 inhibitor组细胞吸光度值、侵袭能力、迁移能力和细胞中E2F1蛋白表达均明显降低,细胞凋亡能力明显升高（P<0.05）;MDA-MB-231组细胞吸光度值、侵袭、迁移、凋亡能力和细胞中E2F1蛋白表达与miR-NC inhibitor组相比差异无统计学意义（P>0.05）。预测软件显示E2F1的3′UTR端与miR-21有碱基互补结合点位。通过向MDA-MB-231细胞中转染野生型E2F1（E2F1-WT）时,miR-21组荧光素酶活性明显低于miR-NC组（P<0.05）;miR-21组和miR-NC组突变体荧光素酶活性相比差异无统计学意义（P>0.05）。与siRNA-NC组相比,siRNA-E2F1组细胞增殖、侵袭、迁移和细胞中E2F1蛋白表达均明显降低,细胞凋亡能力明显增加（P<0.05）。与miR-NC inhibitor组裸鼠移植肿瘤第8天时相比,miR-21 inhibitor组裸鼠肿瘤体积明显降低,肿瘤抑制率为45.3%（P<0.05）。结论低表达miR-21可抑制三阴性乳腺癌细胞增殖、侵袭,促进凋亡,且抑制裸鼠移植瘤体积,其作用机制可能与抑制E2F1表达有关。     

淫羊藿苷对人牙髓干细胞神经向分化作用的初步研究

目的 探讨淫羊藿苷（ICA）对人牙髓干细胞（DPSC）神经向分化的作用。方法 分离培养DPSCs后,采用不同浓度ICA(0.01μmol、0.10μmol、1.00μmol、10.00μmol)处理DPSCs。CCK-8法检测ICA对细胞增殖活力的影响,确定其最佳促DPSCs增殖浓度。分别进行细胞形态学观察和Nestin细胞免疫荧光检测。Western blotting检测DPSCs神经向分化相关蛋白Nestin、βⅢ-tubulin及NSE的表达。结果 不同浓度ICA细胞相对活力值比较,差异有统计学意义（P <0.05）,其中以0.10μmol ICA细胞相对活力最高。与对照组比较,ICA组神经球的直径增加（P <0.05）,Nestin荧光强度增强（P <0.05）,Nestin、βⅢ-tubulin、NSE蛋白相对表达量升高（P <0.05）。结论 ICA能够有效促进DPSCs增殖,提高DPSCs神经向分化能力。     

Plateau potentials evoked by climbing-fibre stimulation are restricted to the Purkinje cell dendrites of the cat

The present study investigated the duration of afterdepolarizations in Purkinje cell somata following climbing-fibre activation. Intracellular recordings revealed that, in cells with membrane potentials more negative than -50 mV and with normal spike-generating capabilities, climbing-fibre activation resulted in somatic responses with short afterdepolarizations. As the cell deteriorated and the resting membrane potential became more positive, the duration and form of the climbing-fibre response resembled the plateau potentials recorded from proximal dendrites. The absence of plateau potentials in undamaged Purkinje cell somata was confirmed by extracellular recording of test spike amplitudes following evoked climbing-fibre responses.     

Bidirectional diode gate: application as a neural spike enhancer

Summary An inexpensive instrument has been described that may be used to eliminate noise in low-level nerve recordings. This electronic manipulation of such signals increases the reliability of digitising or illustrating neural events while eliminating ambiguous noise levels.     

Basolateral junctions are sufficient to suppress epithelial invasion during Drosophila oogenesis.

Epithelial junctions play crucial roles during metazoan evolution and development by facilitating tissue formation, maintenance, and function. Little is known about the role of distinct types of junctions in controlling epithelial transformations leading to invasion of neighboring tissues. Discovering the key junction complexes that control these processes and how they function may also provide mechanistic insight into carcinoma cell invasion. Here, using the Drosophila ovary as a model, we show that four proteins of the basolateral junction (BLJ), Fasciclin-2, Neuroglian, Discs-large, and Lethal-giant-larvae, but not proteins of other epithelial junctions, directly suppress epithelial tumorigenesis and invasion. Remarkably, the expression pattern of Fasciclin-2 predicts which cells will invade. We compared the apicobasal polarity of BLJ tumor cells to border cells (BCs), an epithelium-derived cluster that normally migrates during mid-oogenesis. Both tumor cells and BCs differentiate a lateralized membrane pattern that is necessary but not sufficient for invasion. Independent of lateralization, derepression of motility pathways is also necessary, as indicated by a strong linear correlation between faster BC migration and an increased incidence of tumor invasion. However, without membrane lateralization, derepression of motility pathways is also not sufficient for invasion. Our results demonstrate that spatiotemporal patterns of basolateral junction activity directly suppress epithelial invasion by organizing the cooperative activity of distinct polarity and motility pathways.