A review of the literature on the use of CRISPR/Cas9 gene therapy to treat hepatocellular carcinoma

Noncoding RNAs instruct the Cas9 nuclease to site speifillyl cleave DNA in the CRISPR/Cas9 system.Despite the high incidence of hepatocellular carcinoma(HCC),the patient's outcome is poor.As a result of the emergence of therapeutic resistance in HCC patients,dlinicians have faced difficulties in treating such tumor.In addition,CRISPR/Cas9 screens were used to identify genes that improve the dlinical response of HCC patients.It is the objective of this article to summarize the current understanding of the use of the CRISPR/Cas9 system for the treatment of cancer,with a particular emphasis on HCC as part of the current state of knowledge.Thus,in order to locate recent developments in oncology research,we examined both the Scopus database and the PubMed database.The ability to selectively interfere with gene expression in combinatorial CRISPR/Cas9 screening can lead to the discovery of new effective HCC treatment regimens by combining clinically approved drugs.Drug resistance can be overcome with the help of the CRISPR/Cas9 system.HCC signature genes and resistance to treatment have been uncovered by genome-scale CRISPR activation screening although this method is not without limitations.It has been extensively examined whether CRISPR can be used as a tool for disease research and gene therapy.CRISPR and its applications to tumor research,particularly in HCC,are examined in this study through a review of the literature.

Nrf2 and Her3 co-expression in cholangiocarcinoma:Possible biological pathways for potential therapeutic approach

To the Editor:Cholangiocarcinomas(CCAs)are heterogeneous group of malignancies,encompassing intrahepatic CCA(iCCA),and extrahepatic CCA(eCCA);they are also classified into common hepatic duct cholangiocarcinoma(CHDCCA),choledocus extrapancreatic cholangiocarcinoma(EPCCA)and choledocus intrapancreatic cholangiocarcinoma(IPCCA)and,finally,gallbladder carcinoma(GBCCA).CCAs are relatively uncommon but.

Validation of LPD Scale for the Assessment of Pressure Ulcer at Home

Objectives: This study was designed to test and validate the new LPD scale in a home care setting. The specific objectives are to validate the LPD scale for subjects cared for at home;and to compare LPD to the Braden scale for internal validity. Method: This multicenter, cross-sectional study was conducted in the domestic environment of subjects cared for Home Care services from North to South of Italy. Data collection lasted 8 months, between June 2018 and September 2020, and consisted of the simultaneous compilation of the new LPD, and the Braden scale. Home Care Expert nurses could interface with the recruited subjects and/or caregivers. The parameters considered to validate the new scale were sensitivity (Se), specificity (Sp), positive predictive values (PPV), odds ratio (OR), and the area under the receiver operating characteristic (ROC) curve. Results: Of the 679 recruited subjects, 63.2% were women, and more than 50% did not have a pressure ulcer. 48.2% of the sample aged over 85 years old;69% was affected by multiple disease, and 76.6% took a lot of drugs. 91.6% of the subjects were affected by a partial or total functional dependency. Around 50% of subjects presented double incontinence, and 43% were conscious and collaborated. 85.4% of subjects lived in a healthy environment. The predictive validity parameters showed: Se 77.25%, Sp 84.04%, PPV 91.37%, and the area under the curve (AUC) 0.88% with a confidence interval (CI) 95%. These values mean a moderately accuracy of the test. Conclusions: The new LPD scale has demonstrated a good capacity for identifying the subjects at risk of pressure ulcer and had a better discriminatory power rather than Braden scale.

Liver involvement in patients with COVID-19 infection:A comprehensive overview of diagnostic imaging features

During the first wave of the pandemic,coronavirus disease 2019(COVID-19)infection has been considered mainly as a pulmonary infection.However,different clinical and radiological manifestations were observed over time,including involvement of abdominal organs.Nowadays,the liver is considered one of the main affected abdominal organs.Hepatic involvement may be caused by either a direct damage by the virus or an indirect damage related to COVID-19 induced thrombosis or to the use of different drugs.After clinical assessment,radiology plays a key role in the evaluation of liver involvement.Ultrasonography(US),computed tomography(CT)and magnetic resonance imaging(MRI)may be used to evaluate liver involvement.US is widely available and it is considered the first-line technique to assess liver involvement in COVID-19 infection,in particular liver steatosis and portal-vein thrombosis.CT and MRI are used as second-and third-line techniques,respectively,considering their higher sensitivity and specificity compared to US for assessment of both parenchyma and vascularization.This review aims to the spectrum of COVID-19 liver involvement and the most common imaging features of COVID-19 liver damage.

3D imaging lipidometry in single cell by in-flow holographic tomography

The most recent discoveries in the biochemical field are highlighting the increasingly important role of lipid droplets(LDs)in several regulatory mechanisms in living cells.LDs are dynamic organelles and therefore their complete characterization in terms of number,size,spatial positioning and relative distribution in the cell volume can shed light on the roles played by LDs.Until now,fluorescence microscopy and transmission electron microscopy are assessed as the gold standard methods for identifying LDs due to their high sensitivity and specificity.However,such methods generally only provide 2D assays and partial measurements.Furthermore,both can be destructive and with low productivity,thus limiting analysis of large cell numbers in a sample.Here we demonstrate for the first time the capability of 3D visualization and the full LD characterization in high-throughput with a tomographic phase-contrast flow-cytometer,by using ovarian cancer cells and monocyte cell lines as models.A strategy for retrieving significant parameters on spatial correlations and LD 3D positioning inside each cell volume is reported.The information gathered by this new method could allow more in depth understanding and lead to new discoveries on how LDs are correlated to cellular functions.

湿热环境下LED球泡灯的多物理场耦合仿真

LED产品的工作环境十分复杂,涉及到热、湿气、挥发性化学物质、盐雾等,对产品的可靠性有着重要的影响。针对以往单纯从热场分析LED可靠性的不足,对处于60%相对湿度以及步进温度水平(45°C,85°C和95°C)下的LED球泡灯,采用有限元分析软件ANSYS,结合热-结构模型、湿-结构模型以及热-结构-湿耦合模型,进行多物理场耦合分析。分析得出了受热湿膨胀应力影响而引起变形及分层的部位。同时,以LED球泡灯为实验对象,进行步进加速实验。实验表明,通过多物理场仿真得到的失效部位与实验观察到的结果基本一致,运用多物理场耦合分析LED球泡灯的可靠性是一种有效的方法。

不同结温下大功率LED封装模组的性能分析

对2种大功率LED封装模组的一些性能参数,如正向电压、光通量、相关色温、显色指数和发光效率的热变化特性等进行了实验测试,并进行了相关分析。实验结果表明,大功率LED封装模组的高结温会降低发光效率,过高时会有其他不良影响,因此,需要采用一些有效的散热方法将LED结温控制在合理的范围内。

Lipoprotein-associated phospholipase A2 prognostic role in atherosclerotic complications

Atherosclerosis manifests itself clinically at advanced stages when plaques undergo hemorrhage and/or rupture with superimposed thrombosis, thus abruptly stopping blood supply. Identification of markers of plaque destabilization at a pre-clinical stage is, therefore, a major goal of cardiovascular research. Promising results along this line were provided by studies investigating the lipoprotein-associated phospholipase A2(Lp-PLA2), a member of phospholipase A2 proteins family that plays a key role in the metabolism of pro-inflammatory phospholipids, as oxidized low-density lipoproteins, and in the generation of pro-atherogenic metabolites, including lysophosphatidylcholine and oxidized free fatty acids. We herein review the experimental and clinical studies supporting use of Lp-PLA2 activity for predicting cardiovascular events. To his end we considered not only Lp-PLA2 activity and mass, but also Lp-PLA2 gene variations and their association with incident coronary artery disease, stroke, and cardiovascular mortality. Based on these evidences the major scientific societies have included in their guidelines the measurement of Lp-PLA2 activity among the biomarkers that are useful in risk stratification of adult asymptomatic patients at intermediate cardiovascular risk. The results of two recently published major clinical trials with the LpPLA2 inhibitor darapladib, which seem to challenge the pathogenic role of Lp-PLA2, will also be discussed.

Intraocular inflammation after Ultrasound Cyclo Plasty for the treatment of glaucoma

This is a prospective interventional clinical study evaluating intraocular inflammation developed after Ultrasound Cyclo Plasty(UCP) for the treatment of glaucoma. Eighteen eyes of 18 patients were treated with UCP second-generation probes(Eye OP1). After treatment, the mean intraocular pressure(IOP) significantly decreased from 26.8±7.2 to 18.8±6.1 mm Hg at day 1 and to 14.7±3.4 mm Hg at month 6(all P<0.001). Mean laser flare-cell photometry value steeply increased after surgery from 12.1±7.5 to 64.1±53.9 ph/ms(P=0.001) at day 1, and then progressively decreased to respectively 60.6±49.7 at day 7, 43.5±38.5 at day 14 and 28.2±18.3 at month 1(all P<0.05), returning at levels similar to baseline ones at month 3 and month 6(respectively 16.7±6.2 and 12.8±10.2, both P>0.05). A significant negative correlation was found between postoperative increase of aqueous flare values and anterior chamber depth(R=-0.568, P=0.014). This timeframe may be considered reasonable for repeating UCP treatment, when required.

Resilience Against Replay Attacks:A Distributed Model Predictive Control Scheme for Networked Multi-Agent Systems

In this paper,a resilient distributed control scheme against replay attacks for multi-agent networked systems subject to input and state constraints is proposed.The methodological starting point relies on a smart use of predictive arguments with a twofold aim:1)Promptly detect malicious agent behaviors affecting normal system operations;2)Apply specific control actions,based on predictive ideas,for mitigating as much as possible undesirable domino effects resulting from adversary operations.Specifically,the multi-agent system is topologically described by a leader-follower digraph characterized by a unique leader and set-theoretic receding horizon control ideas are exploited to develop a distributed algorithm capable to instantaneously recognize the attacked agent.Finally,numerical simulations are carried out to show benefits and effectiveness of the proposed approach.

预防保健：围产期妇幼保健中缺失的一环

1 The need to extend and expand the maternal,neonatal and child health (MNCH) continuum to include preconception health The provision of care along the MNCH continuum is currently the core strategy for reducing maternal,neonatal and infant mortality [1-5]. Yet,gaps in the MNCH continuum persist. Reductions in maternal mortality have stagnated in many parts of the world. With the lowering of infant and child mortality,approximately 40% of infant deaths now occur in the neonatal period [6]. Strong scientific evidence indicates that improving women's health before pregnancy would result in better maternal and neonatal health outcomes-specifically,fewer pregnancy complications and a reduction in rates of birth defects,fetal loss,low birth weight and preterm delivery [7]. The WHO estimates that 19% of all maternal deaths are due indirectly to preexisting medical conditions [8]. Furthermore,early prenatal care is too late to prevent many adverse birth outcomes,as organogenesis is completed during the critical first weeks of pregnancy,often before a woman knows that she is pregnant [9]. There is a clear need to strengthen preconception health services.

用阶梯状掺杂埋层对超浅结进行C-V剖面分析(英文)

研究了应用双边C-V法测量超浅结(如p+-n结)的掺杂分布。推导了在已知p+-n结的电容-电压(C-VR)关系、n区掺杂、以及热平衡下n区耗尽层宽度(xn0)的情况下计算p区掺杂浓度分布的公式。xn0是计算p区掺杂分布所需的一个关键参数,通过将n区掺杂设计成阶梯状,可实现对xn0的精确提取。用Medici对具有相同的阶梯状掺杂n区的p+-n和n-肖特基结进行器件仿真可得其C-VR关系。运用常规C-V法,由肖特基结的C-VR关系可提取出n区掺杂浓度。实现了对xn0的精确提取,其精度达1.8nm。基于精确的xn0,运用双边C-V法提取的p+区的掺杂浓度分布与Medici仿真结果非常吻合。

A Resilient Control Strategy for Cyber-Physical Systems Subject to Denial of Service Attacks:A Leader-Follower Set-Theoretic Approach

Multi-agent systems are usually equipped with open communication infrastructures to improve interactions efficiency,reliability and sustainability.Although technologically costeffective,this makes them vulnerable to cyber-attacks with potentially catastrophic consequences.To this end,we present a novel control architecture capable to deal with the distributed constrained regulation problem in the presence of time-delay attacks on the agents’communication infrastructure.The basic idea consists of orchestrating the interconnected cyber-physical system as a leader-follower configuration so that adequate control actions are computed to isolate the attacked unit before it compromises the system operations.Simulations on a multi-area power system confirm that the proposed control scheme can reconfigure the leader-follower structure in response to denial ofservice(DoS)attacks.

Autoimmune gastritis:Pathologist's viewpoint

Western countries are seeing a constant decline in the incidence of Helicobacter pylori-associated gastritis, coupled with a rising epidemiological and clinical impact of autoimmune gastritis. This latter gastropathy is due to autoimmune aggression targeting parietal cells through a complex interaction of auto-antibodies against the parietal cell proton pump and intrinsic factor, and sensitized T cells. Given the specific target of this aggression, autoimmune gastritis is typically restricted to the gastric corpus-fundus mucosa. In advanced cases, the oxyntic epithelia are replaced by atrophic(and metaplastic) mucosa, creating the phenotypic background in which both gastric neuroendocrine tumors and(intestinal-type) adenocarcinomas may develop. Despite improvements in our understanding of the phenotypic changes or cascades occurring in this autoimmune setting, no reliable biomarkers are available for identifying patients at higher risk of developing a gastric neoplasm. The standardization of autoimmune gastritis histology reports and classifications in diagnostic practice is a prerequisite for implementing definitive secondary prevention strategies based on multidisciplinary diagnostic approaches integratingendoscopy, serology, histology and molecular profiling.

Potential advantages of acute kidney injury management by mesenchymal stem cells

Mesenchymal stem cells are currently considered as a promising tool for therapeutic application in acute kidney injury(AKI) management. AKI is characterized by acute tubular injury with rapid loss of renal function. After AKI, inflammation, oxidative stress and excessive deposition of extracellular matrix are the molecular events that ultimately cause the end-stage renal disease. Despite numerous improvement of supportive therapy, the mortality and morbidity among patients remain high. Therefore, exploring novel therapeutic options to treat AKI is mandatory. Numerous evidence in animal models has demonstrated the capability of mesenchymal stem cells(MSCs) to restore kidney function after induced kidney injury. After infusion, MSCs engraft in the injured tissue and release soluble factors and microvesicles that promote cell survival and tissue repairing. Indeed, the main mechanism of action of MSCs in tissue regeneration is the paracrine/endocrine secretion of bioactive molecules. MSCs can be isolated from several tissues, including bone marrow, adipose tissue, and blood cord; pre-treatment procedures to improve MSCs homing and their paracrine function have been also described. This review will focus on the application of cell therapy in AKI and it will summarize preclinical studies in animal models and clinical trials currently ongoing about the use of mesenchymal stem cells after AKI.

HER2 heterogeneity in gastric/gastroesophageal cancers: From benchside to practice

HER2 is overexpressed in approximately 10%-20% of gastric and gastroesophageal junction carcinomas. In these types of cancer, accurate assessment of HER2 status is mandatory, for selecting patients who may benefit from targeted therapies with anti-HER2 drugs such as Trastuzumab. This manuscript focuses on HER2 in gastric carcinogenesis, on optimal evaluation of HER2 and on the possible causes which may contribute to inaccurate HER2 evaluation. Similarly to breast cancer HER2 evaluation, standardization of HER2 testing in gastric cancer is necessary in diagnostic practice. The three principle aspects which require consideration are:(1) the choice of sample with regards to cancer morphology- intestinal vs diffuse areas;(2) the choice of scoring criteria- use of HER2 scoring criteria specific for gastric cancer; and(3) the choice of HER2 evaluation methods- use of an algorithm in which both immunohistochemistry and in situ hybridization play a role. Problematic issues include:(1) pre-analytic variables with particular emphasis on fixation;(2) recommended methodology for HER2 assessment(immunohistochemistry vs in situ hybridization);(3) HER2 heterogeneity both within the primary tumor and between primary tumor and metastases;(4) reliability of biopsies in HER 2 evaluation; and(5) quantity of sample(FFPE blocks from surgical specimens or endoscopic biopsies) necessary for an adequate assessment.

role of microRNAs in the main molecular pathways of hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is the most common primary liver malignant neoplasia. HCC is characterized by a poor prognosis. The need to find new molecular markers for its diagnosis and prognosis has led to a progressive increase in the number of scientific studies on this topic. MicroRNAs(miRNAs) are small noncoding RNA that play a role in almost all main cellular pathways. miRNAs are involved in the regulation of expression of the major tumor-related genes in carcinogenesis, acting as oncogenes or tumor suppressor genes. The aim of this review was to identify papers published in 2017 investigating the role of miRNAs in HCC tumorigenesis. miRNAs were classified according to their role in the main molecular pathways involved in HCC tumorigenesis:(1) m TOR;(2) Wnt;(3) JAK/STAT;(4) apoptosis; and(5) MAPK. The role of miRNAs in prognosis/response prediction was taken into consideration. Bearing in mind that the analysis of miRNAs in serum and other body fluids would be crucial for clinical management, the role of circulating miRNAs in HCC patients was also investigated. The most represented mi RNA-regulated pathway in HCC is m TOR, but apoptosis, Wnt, JAK/STAT or MAPK pathways are also influenced by mi RNA expression levels. These miRNAs could thus be used in clinical practice as diagnostic, prognostic or therapeutic targets for HCC treatment.

Physical energies to the rescue of damaged tissues

Rhythmic oscillatory patterns sustain cellular dynamics, driving the concerted action of regulatory molecules, microtubules, and molecular motors. We describe cellular microtubules as oscillators capable of synchronization and swarming, generating mechanical and electric patterns that impact biomolecular recognition. We consider the biological relevance of seeing the inside of cells populated by a network of molecules that behave as bioelectronic circuits and chromophores. We discuss the novel perspectives disclosed by mechanobiology, bioelectromagnetism, and photobiomodulation, both in term of fundamental basic science and in light of the biomedical implication of using physical energies to govern (stem) cell fate. We focus on the feasibility of exploiting atomic force microscopy and hyperspectral imaging to detect signatures of nanomotions and electromagnetic radiation (light), respectively, generated by the stem cells across the specification of their multilineage repertoire. The chance is reported of using these signatures and the diffusive features of physical waves to direct specifically the differentiation program of stem cells in situ, where they already are resident in all the tissues of the human body. We discuss how this strategy may pave the way to a regenerative and precision medicine without the needs for (stem) cell or tissue transplantation. We describe a novel paradigm based upon boosting our inherent ability for self-healing.

Epilepsy associated tumors: Review article

Long-term epilepsy associated tumors(LEAT) represent a well known cause of focal epilepsies. Glioneuronaltumors are the most frequent histological type consisting of a mixture of glial and neuronal elements and most commonly ariseing in the temporal lobe. Cortical dysplasia or other neuronal migration abnormalities often coexist. Epilepsy associated with LEAT is generally poorly controlled by antiepileptic drugs while, on the other hand, it is high responsive to surgical treatment. However the best management strategy of tumor-related focal epilepsies remains controversial representing a contemporary issues in epilepsy surgery. Temporo-mesial LEAT have a widespread epileptic networkwith complex epileptogenic mechanisms. By using an epilepsy surgery oriented strategy LEAT may have an excellent seizure outcome therefore surgical treatment should be offered early, irrespective of pharmacoresistance, avoiding both the consequences of uncontrolled seizures as well as the side effects of prolonged pharmacological therapy and the rare risk of malignant transformation.

Twenty-year survival after iterative surgery for metastatic renal cell carcinoma: A case report and review of literature

BACKGROUND The therapeutic approach of metastatic renal cell carcinoma(RCC)represents a real challenge for clinicians,because of the variable clinical course;the recent availability of numerous targeted therapies that have significantly improved overall oncological results,but still with a low percentage of complete responses;and the increasing role of metastasectomy(MSX)as an effective strategy to achieve a durable cure,or at least defer initiation of systemic therapies,in selected patients and in the context of multimodality treatment strategies.CA^E SUMMARY We report here the case of a 40-year-old man who was referred to our unit in November 2004 with lung and mediastinal lymph nodes metastases identified during periodic surveillance 6 years after a radical nephrectomy for RCC;he underwent MSX of multiple lung nodules and mediastinal lymphadenectomy,with subsequent systemic therapy with Fluorouracil,Interferon-alpha and Interleukin 2.The subsequent clinical course was characterized by multiple sequential abdominal and thoracic recurrences,successfully treated with multiple systemic treatments,repeated local treatments,including two pancreatic resections,conservative resection and ablation of multiple bilobar liver metastases,resection and stereotactic body radiotherapy of multiple lung metastases.He is alive without evidence of recurrence 20 years after initial nephrectomy and sequential treatment of recurrences in multiple sites,including resection of more than 38 metastases,and 5 years after his last MSX.CONCLUSION This case highlights that effective multimodality therapeutic strategies,including multiple systemic treatments and iterative aggressive surgical resection,can be safely performed with long-term survival in selected patients with multiple metachronous sequential metastases from RCC.