Bench to bedside: recent advances in lung cancer pathological research with implications for diagnostic clinical practice

After decades of relative stagnation lung cancer is emerging as a disease type where rapid progress is being made in diagnosis and therapy, as well as in our understanding of disease biology. Much of this progress is of immediate impact to diagnosticians, and more is likely to affect diagnostic practice in the near future. In this review we seek to briefly summarize several key areas of active research of immediate or probable imminent value to trainee and consultant pulmonary pathologists alike. We cover some major changes in tumour classification, grading, and patient stratification, as well as considering the state of the art in machine-assisted interpretation of lung cancer histology, and the use of genetically modified lung cancer models.     

丹参F3'5'H基因克隆及其序列分析

目的:为了验证丹参类黄酮-3′,5′-羟基化酶(Flavonoid 3′,5′-hydroxylase,F3′5′H)基因与丹参花色表型的相关性,本研究克隆并分析了紫花丹参99-3株系和一些白花丹参中的F3′5′H基因。方法:本研究通过提取紫花丹参和白花丹参中的总RNA,再将其反转录得到cDNA,以此cDNA为模板,利用PCR方法扩增获得F3′5′H基因全长序列。再利用生物信息学分析方法,分析了该基因编码蛋白质的理化性状、结构域、系统进化等特点,并预测了该蛋白质的亚细胞定位、跨膜区等;利用实时定量PCR方法检测了该基因的表达特异性;利用毛状根遗传转化方法获得了该基因的过表达阳性毛状根株系。结果:F3′5′H基因全长1551 bp,编码516个氨基酸,相对分子质量为57.4 kDa。该基因在丹参花中高丰度表达。在42份(紫花25份;白花17份)丹参样品中发现一个单核苷酸多态性(Single Nucleotide Polymorphisms,SNP)位点在紫花和白花丹参中呈现与花色相关的稳定变化。系统发育分析表明丹参F3′5′H与美女樱的F3′5′H具有较高序列同源性。获得了过表达F3′5′H的毛状根株系。结论:本研究在丹参中克隆到F3′5′H基因,对其序列进行了分析,为进一步研究丹参F3′5′H基因的功能奠定基础。     

Benzophenone-3 promotion of mammary tumorigenesis is diet-dependent

Benzophenone-3 is a putative endocrine disrupting chemical and common ingredient in sunscreens. The potential of endocrine disrupting chemicals to act as agonists or antagonists in critical hormonally regulated processes, such as mammary gland development and mammary tumorigenesis, demands evaluation of its potential in promoting breast cancer. This study identifies the effects of BP-3 on mammary tumorigenesis with high-fat diet during puberty versus adulthood in Trp53-null transplant BALB/c mice. Benzophenone-3 exposure yielded levels in urine similar to humans subjected to heavy topical sunscreen exposure. Benzophenone-3 was protective for epithelial tumorigenesis in mice fed lifelong low-fat diet, while promotional for epithelial tumorigenesis in mice fed adult high-fat diet. Benzophenone-3 increased tumor cell proliferation, decreased tumor cell apoptosis, and increased tumor vascularity dependent on specific dietary regimen and tumor histopathology. Even in instances of an ostensibly protective effect, other parameters suggest greater risk. Although benzophenone-3 seemed protective on low-fat diet, spindle cell tumors arising in these mice showed increased proliferation and decreased apoptosis. This points to a need for further studies of benzophenone-3 in both animal models and humans as a potential breast cancer risk factor, as well as a more general need to evaluate endocrine disrupting chemicals in varying dietary contexts.     

Avian mud nest architecture by self-secreted saliva

Mud nests built by swallows (Hirundinidae) and phoebes (Sayornis) are stable granular piles attached to cliffs, walls, or ceilings. Although these birds have been observed to mix saliva with incohesive mud granules, how such biopolymer solutions provide the nest with sufficient strength to support the weight of the residents as well as its own remains elusive. Here, we elucidate the mechanism of strong granular cohesion by the viscoelastic paste of bird saliva through a combination of theoretical analysis and experimental measurements in both natural and artificial nests. Our mathematical model considering the mechanics of mud nest construction allows us to explain the biological observation that all mud-nesting bird species should be lightweight.

Bird nests come in a variety of forms made from diverse building materials (1, 2). Each type of bird nest is subjected to mechanical constraints imposed by material characteristics. To overcome these constraints, birds have devised brilliant architectural technologies, which provide inspiration for a novel materials processing scheme and help us to better understand animal behavior.For instance, some birds including storks (Cicioniidae) and eagles (Accipitidae) build nests by piling up hard filamentary materials such as twigs, harnessing their friction as the cohesion mechanism (3). Weaverbirds (Ploceidae) weave soft filamentary materials such as grass and fine leaves into a woven nest tied to a tree branch. Some bird species use their own saliva in nest building, which Darwin considered an example of natural selection (4). An extreme case is the Edible-nest Swiftlets, which build their nest purely of self-secreted saliva so that it can be attached to cliff walls and cave ceilings where the above twig piles and tied leaves are not allowed (5).Swallows (Hirundinidae), phoebes (Sayornis), and other mud nesters have developed a unique building material, a mixture of mud and their own saliva, in contrast to those made of purely collected or self-secreted materials (6) (Fig. 1). During construction, mud nesters repeatedly pile a beakful of wet mud on the nest, and liquid bridges are formed in the nest due to evaporation. While building a nest usually takes several weeks, a transition from wet to dry structures can occur within a few hours. Hence, the capillary forces of liquid bridges temporarily provide cohesion such as those in sandcastles. However, unlike sandcastles, dehydrated saliva comes into play for permanent cohesion after complete evaporation (SI Appendix, Supplementary Note 1).Open in a separate windowFig. 1.A nest of the barn swallow (H. rustica). (A) Photograph of a barn swallow nest, taken from under the ceiling of a house in Suwon-si, Gyunggi-do, South Korea (37°16′13.5″N 126°59′01.0″E). (B) SEM image of the nest surface. (C) Chemical composition analysis of the surface shown in B by EDS. The red area indicates a region containing mostly carbon atoms, which may originate from bird saliva. The green area indicates a region containing mostly the silicon atoms of clay particles.Mud itself cannot confer sufficient cohesion and adhesion in mud nests. The ability of mud nests to bear tensile loads originates from the gluing agent in the bird''s saliva, which permeates into granules as a liquid and binds them as a solid after solvent evaporation (6–8) (SI Appendix, Supplementary Note 2). The gluing agent is called mucin, a family of large glycoproteins that are ubiquitous in animal organs and form a mucus gel with versatile functionality (9). Fig. 1B shows the scanning electron microscopy (SEM) image of a barn swallow’s mud nest consisting of platelet clay particles and larger grains. Energy-dispersive spectroscopy (EDS) mapping image of Fig. 1C clearly shows regions corresponding to organic material which is presumed to be from bird’s saliva.Of particular interest and worth biophysical investigation are the tensile strength of the mud nest with hardened saliva, design principles associated with the saliva-originated strength, and the resulting effects on the evolution of these mud-nesting birds. Principles behind cohesion in granular materials, such as wet sands (10), cemented powder aggregates (11), construction materials (12), and pharmaceutical tablets (13), have been studied to date, exploring the stress transmission, elasticity, and failure (14–18), and the formation of solidified bridges (19–21). However, little attention has been paid to the cohesion effects of self-secreted polymer materials upon evaporation and the biologically constructed granular architecture like birds’ mud nests. Here we devised experimental techniques to measure the strength of the relatively small and fragile nest specimens in order to mechanically characterize birds’ mud nests. We elucidate how solutes from bird saliva generate solid bridges that give rise to macroscopic tensile strength, which has long awaited physicochemical explanation since its first observation (4). To characterize the design principle of bird''s mud nests, we investigated natural and three-dimensional (3D)-printed artificial nests with various tools for visualization and mechanical testing. Along with the experimental studies, we theoretically investigated the effects of biopolymer concentration on nest strength. This combination of theory and experiment suggests that there is a size limit for mud-nesting birds, which is supported by biological data.     

Using cortical non-invasive neuromodulation as a potential preventive treatment in schizophrenia - A review

BackgroundEvidence suggests that schizophrenia constitutes a neurodevelopmental disorder, characterized by a gradual emergence of behavioral and neurobiological abnormalities over time. Therefore, applying early interventions to prevent later manifestation of symptoms is appealing.ObjectiveThis review focuses on the use of cortical neuromodulation in schizophrenia and its potential as a preventive treatment approach. We present clinical and preclinical findings investigating the use of neuromodulation in schizophrenia, including the current research focusing on cortical non-invasive stimulation and its possibility as a future preventive treatment.MethodsWe performed a search in Medline (PubMed) in September 2020 using a combination of relevant medical subject headings (MeSH) and text words. The search included human and preclinical trials as well as existing systematic reviews and meta-analysis. There were no restrictions on language or the date of publication.ResultsNeurodevelopmental animal models may be used to investigate how the disease progresses and thus which brain areas ideally should be targeted at a given time point. Here, abnormalities of the prefrontal cortex have been often identified as an early and persistent impairment in schizophrenia. Currently there is insufficient evidence to either support or refute the use of neuromodulation to the cortex in adult patients with already manifested symptoms. However, preclinical results show that early non-invasive neuromodulation to the prefrontal cortex of adolescent animals, sufficiently prevents later psychosis-relevant abnormalities in adulthood. This points to the promising potential of cortical non-invasive neuromodulation as a preventive treatment when applied early in the course of the disease.ConclusionPreclinical translational-oriented findings indicate, that neuromodulation to cortical areas offers the possibility of targeting early neuropathology and through this diminish the progression of a later schizophrenic profile. Further studies are needed to investigate whether such early cortical stimulation may serve as a future preventive treatment in schizophrenia.     

Modelling human pathology of traumatic brain injury in animal models

Traumatic brain injury (TBI) is caused by a head impact with a force exceeding regular exposure from normal body movement which the brain normally can accommodate. People affected include, but are not restricted to, sport athletes in American football, ice hockey, boxing as well as military personnel. Both single and repetitive exposures may affect the brain acutely and can lead to chronic neurodegenerative changes including chronic traumatic encephalopathy associated with the development of dementia. The changes in the brain following TBI include neuroinflammation, white matter lesions, and axonal damage as well as hyperphosphorylation and aggregation of tau protein. Even though the human brain gross anatomy is different from rodents implicating different energy transfer upon impact, especially rotational forces, animal models of TBI are important tools to investigate the changes that occur upon TBI at molecular and cellular levels. Importantly, such models may help to increase the knowledge of how the pathologies develop, including the spreading of tau pathologies, and how to diagnose the severity of the TBI in the clinic. In addition, animal models are helpful in the development of novel biomarkers and can also be used to test potential disease‐modifying compounds in a preclinical setting.     

全身振动训练联合正弦交变电磁场对去卵巢骨质疏松大鼠骨密度、骨代谢指标的影响研究

目的探讨全身振动训练联合正弦交变电磁场对去卵巢骨质疏松大鼠骨密度、骨代谢、骨生物力学性能的影响。方法将100只大鼠随机分为切除卵巢组和对照组,分别进行卵巢切除术和假手术,大鼠进行6周恢复,恢复后将造模成功的切除卵巢大鼠随机分为模型组(MO组)、全身振动训练组(W组)、正弦交变电磁场组(S组)、全身振动训练+正弦交变电磁场组(WS组),对照组列为假手术组(SO组),进行为期16周干预,干预结束后对大鼠进行骨密度、骨代谢、骨生物力学性质的检测。结果16周干预完成后,MO组、S组大鼠体质量显著高于SO组、W组、WS组大鼠(P0. 05); W组、WS组、SO组大鼠骨密度指标、血清雌二醇浓度指标明显高于大鼠MO组(P0. 05); SO组大鼠血清雌二醇浓度指标明显高于S组、W组、WS组(P0. 05); M组大鼠血清OC、ALP浓度明显高于SO组、W组、S组、WS组大鼠(P0. 05);尿液DPD/Cre、Ca/Cre、P/Cre浓度方面,M组大鼠明显高于SO组、W组、S组、WS组大鼠(P0. 05)。SO组、W组、S组、WS组大鼠股骨最大载荷和弹性模型明显高于MO组大鼠(P0. 05)。SO组、W组、S组、WS组大鼠股骨断裂载荷组间无明显差异(P0. 05); SO组、W组、S组、WS组大鼠股骨弹性模量明显高于MO组大鼠(P0. 05)。SO组大鼠股骨弹性模量明显高于S组大鼠(P0. 05),但与W组、WS组大鼠没有明显差异(P0. 05)。L4椎体压缩试验,SO组、W组、S组、WS组大鼠股骨最大载荷和弹性模型明显高于MO组大鼠(P0. 05)。SO组大鼠高于S组、W组,差异有统计学意义(P0. 05),但与WS组大鼠没有明显差异(P0. 05)。结论全身振动训练、正弦交变电磁场、全身振动训练联合正弦交变电磁场3种干预方式施用于去卵巢骨质疏松大鼠均能提升骨密度、抑制骨吸收、平衡骨代谢、改善骨骼结构力学和材料力学性能。而全身振动训练联合正弦交变电磁场能的治疗效果优于单纯使用全身振动训练或正弦交变电磁场,在临床应用中有一定推广价值。     

腰椎间盘突出症动物模型的研究进展

近年来,腰椎间盘突出症发病率逐渐增高,由于动物模型对于阐明疾病发病机制及评估新的治疗方法具有重要作用,越来越多的学者开始研究如何建立合适的、能够更好地模拟人类腰椎间盘突出的动物模型。近期的研究多选用包括大鼠、兔、羊、猪等动物作为实验对象,通过物理或化学方式直接损伤其椎间盘或神经根,或通过限制实验动物的行为动作、改变其饲养环境以及性别年龄导致的生理因素等方式诱导其出现椎间盘退变。各种造模方法均有其特点及局限性,适用于不同情况,有必要继续完善。     

Utility of food pellets containing 1-aminobenzotriazole for longer term in vivo inhibition of cytochrome P450 in mice

1. The utility of 1-aminobenzotriazole (ABT), incorporated in food, has been investigated as an approach for longer term inhibition of cytochrome P450 (P450) enzymes in mice.

2. In rats, ABT inhibits gastric emptying, to investigate this potential limitation in mice we examined the effect of ABT administration on the oral absorption of NVS-CRF38. Two hour prior oral treatment with 100?mg/kg ABT inhibited the oral absorption of NVS-CRF38, T_max was 4?hours for ABT-treated mice compared to 0.5?hours in the control group.

3. A marked inhibition of hepatic P450 activity was observed in mice fed with ABT containing food pellets for 1?month. P450 activity, as measured by the oral clearance of antipyrine, was inhibited on day 3 (88% of control), week 2 (83% of control) and week 4 (80% of control).

4. T_max values for antipyrine were comparable between ABT-treated mice and the control group, alleviating concerns about impaired gastric function.

5. Inclusion of ABT in food provides a minimally invasive and convenient approach to achieve longer term inhibition of P450 activity in mice. This model has the potential to enable pharmacological proof-of-concept studies for research compounds which are extensively metabolised by P450 enzymes.