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The applicability of electrical impedance tomography (EIT) in birds is unknown. This study aimed to evaluate the use of EIT in anaesthetised chickens in four recumbency positions. Four adult Hyline chickens were anaesthetised with isoflurane in oxygen, and intubated endotracheally for computed tomography (CT). A rubber belt was placed around the coelom caudal to the shoulder joint. A chicken-specific finite element (FE) model, which is essential to generate anatomically accurate functional EIT images for analysis, was constructed based on the CT images obtained at the belt level. Ten additional chickens were anaesthetised with the same protocol. An EIT electrode belt was placed at the same location. The chickens were breathing spontaneously and positioned in dorsal, ventral, right and left lateral recumbency in a randomised order. For each recumbency, raw EIT data were collected over 2 min after 13 min of stabilisation. The data were reconstructed into functional EIT images. EIT variables including tidal impedance variation (TIV), centre of ventilation right to left (CoVRL) and ventral to dorsal (CoVVD), right to left (RL) ratio, impedance change (ΔZ) and eight regional impedance changes including the dorsal, central-dorsal, central-ventral and ventral regions of the right and left regions were analysed. Four breathing patterns (BrP) were observed and categorised based on the expiratory curve. A linear mixed model was used to compare EIT variables between recumbencies. Fisher's exact test was used to compare the frequencies of breathing patterns for each recumbency. The ΔZ observed was synchronous to ventilation, and represented tidal volume of the cranial air sacs as confirmed by CT. Significant differences were found in CoVVD and regional impedance changes between dorsal and ventral recumbencies (P < 0.05), and in CoVRL, RL ratio and regional impedance changes between right and left recumbencies (P < 0.05), which suggested a tendency for the distribution of ventilation to shift towards non-dependent air sacs. No differences were found for TIV and respiratory rate between recumbencies. Recumbency had a significant effect on the frequencies of each of the four BrPs (P = 0.001). EIT can monitor the magnitude and distribution of ventilation of the cranial air sacs in different recumbencies in anaesthetised chickens.

Yu Hei,1 Binhong Teng,2,3 Ziqian Zeng,2,3 Siqi Zhang,3 Qian Li,3 Jijia Pan,3 Zuyuan Luo,3 Chunyang Xiong,1 Shicheng Wei2,3 1Department of Mechanics and Engineering Science, College of Engineering, Peking University, Beijing, People’s Republic of China; 2Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Peking University, Beijing, People’s Republic of China; 3Laboratory of Biomaterials and Regenerative Medicine, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, People’s Republic of ChinaCorrespondence: Chunyang XiongDepartment of Mechanics and Engineering Science, College of Engineering, Peking University, Beijing, People’s Republic of ChinaTel +86 10 62757940Email cyxiong@pku.edu.cnShicheng WeiDepartment of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Peking University, Beijing, People’s Republic of ChinaTel +86 10 82195780Email sc-wei@pku.edu.cnBackground: Immune checkpoint blockades (ICBs) are a promising treatment for cancers such as melanoma by blocking important inhibitory pathways that enable tumor cells to evade immune attack. Programmed death ligand 1 monoclonal antibodies (aPDL1s) can be used as an ICB to significantly enhance the effectiveness of tumor immunotherapy by blocking the PD-1/PD-L1 inhibitory pathway. However, the effectiveness of aPDL1s may be limited by low selectivity in vivo and immunosuppressed tumor microenvironment including hypoxia.Purpose: To overcome the limitations, we develop a multifunctional immunoliposome, called CAT@aPDL1-SSL, with catalase (CAT) encapsulated inside to overcome tumor hypoxia and aPDL1s modified on the surface to enhance immunotherapeutic effects against melanoma.Methods: The multifunctional immunoliposomes (CAT@aPDL1-SSLs) are prepared using the film dispersion/post-insertion method. The efficacy of CAT@aPDL1-SSLs is verified by multiple experiments in vivo and in vitro.Results: The results of this study suggest that the multifunctional immunoliposomes preserve and protect the enzyme activity of CAT and ameliorate tumor hypoxia. Moreover, the enhanced cellular uptake of CAT@aPDL1-SSLs in vitro and their in vivo biodistribution suggest that CAT@aPDL1-SSLs have great targeting ability,resulting in improved delivery and accumulation of immunoliposomes in tumor tissue.Finally, by activating and increasing the infiltration of CD8+ T cells at the tumor site, CAT@aPDL1-SSLs inhibit the growth of tumor and prolong survival time of mice,with low systemic toxicity.Conclusion: In conclusion, the multifunctional immunoliposomes developed and proposed in this study are a promising candidate for melanoma immunotherapy, and could potentially be combined with other cancer therapies like radiotherapy and chemotherapy to produce positive outcomes.Keywords: immunotherapy, programmed death ligand 1 monoclonal antibodies, aPDL1s, tumor hypoxia, melanoma, liposomes

Yuan Ding,1,* Wei Cui,2,* Dan Sun,1 Gui-Ling Wang,1 Yu Hei,1 Shuai Meng,1 Jian-Hua Chen,3 Ying Xie,1 Zhi-Qiang Wang4 1Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems, Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, 2School of Chemistry and Chemical Engineering, University of Chinese Academy of Sciences, Beijing, 3School of Medicine, Jianghan University, Wuhan, People’s Republic of China; 4Department of Chemistry and Biochemistry, Kent State University Geauga, Burton, OH, USA *These authors contributed equally to this work Abstract: In vivo evaluation of drug delivery vectors is essential for clinical translation. In BALB/c nude mice bearing human breast cancer tumors, we investigated the biocompatibility, pharmacokinetics, and pharmacodynamics of doxorubicin (DOX)-loaded novel cell-penetrating peptide (CPP)-modified pH-sensitive liposomes (CPPL) (referred to as CPPL(DOX)) with an optimal CPP density of 4%. In CPPL, a polyethylene glycol (PEG) derivative formed by conjugating PEG with stearate via acid-degradable hydrazone bond (PEG2000-Hz-stearate) was inserted into the surface of liposomes, and CPP was directly attached to liposome surfaces via coupling with stearate to simultaneously achieve long circulation time in blood and improve the selectivity and efficacy of CPP for tumor targeting. Compared to PEGylated liposomes, CPPL enhanced DOX accumulation in tumors up to 1.9-fold (p

Chang Liu,1,* Xiao-Na Liu,1,* Gui-Ling Wang,1 Yu Hei,1 Shuai Meng,1 Ling-Fei Yang,1 Lan Yuan,2 Ying Xie1 1Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems, Department of Pharmaceutics, School of Pharmaceutical Sciences, 2Medical and Healthy Analysis Center, Peking University, Beijing, People’s Republic of China *These authors contributed equally to this work Abstract: As the global population ages, cancer rates increase worldwide, and degenerative diseases of the central nervous system (CNS), brain tumors, and inflammation threaten human health more frequently. We designed a dual-mediated (receptor-mediated and adsorption-mediated) liposome, named transferrin–cell penetrating peptide–sterically stabilized liposome (TF-CPP-SSL), to improve therapy for gliomas through combining molecular recognition of transferrin receptors (TF-Rs) on the blood–brain barrier (BBB) and glioma cells with the internalization and lysosomal escaping ability of CPP. Based on the systematic investigation of structure–activity relations on the cellular level, we constructed TF-CPP-SSL rationally by conjugating TF and CPP moieties to the liposomes via PEG3.4K and PEG2.0K, respectively, and found the optimum densities of TF and CPP were 1.8% and 4%, respectively. These liposomes had the highest targeting efficacy for brain microvascular endothelial cell and C6 cell uptake but avoided capture by normal cells. Fluorescence resonance energy transfer technology and coculture models of BBB and glioma C6 cells indicated that TF-CPP-SSL was transported across the BBB without drug leakage, liposome breakup, or cleavage of ligand. TF-CPP-SSL offered advantages for crossing the BBB and entering into glioma C6 cells. Real-time confocal viewing revealed that TF-CPP-SSL was entrapped in endosomes of glioma C6 cells and then escaped from lysosomes successfully to release the liposomal contents into the cytosol. Entrapped contents, such as doxorubicin, could then enter the nucleus to exert pharmacological effects. Keywords: dual ligand-modified liposomes, transferrin, cell-penetrating peptide, real-time confocal, fluorescence resonance energy transfer, tumor targeting

Karynsa Cetin1, David S Ettinger2, Yong-jiang Hei3, Cynthia D O'Malley11Center for Observational Research, Amgen Inc., Thousand Oaks, CA, USA; 2Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA; 3Global Development, Amgen Inc., Thousand Oaks, CA, USABackground: The role of histology in the targeted management of nonsmall cell lung cancer (NSCLC) has garnered renewed attention in recent years. We provide contemporary population-based estimates of survival and an assessment of important prognostic factors in stage IV NSCLC by major histologic subtype.Methods: Using data from the Surveillance, Epidemiology and End Results (SEER) Program, we stratified 51,749 incident stage IV NSCLC patients (1988–2003 with follow-up through 2006) by major histologic subtype. We used Kaplan–Meier and Cox proportional hazards methods to describe overall survival and the prognostic influence of select patient, tumor, and treatment characteristics for each histologic subgroup.Results: Survival was highest in patients with bronchioloalveolar adenocarcinoma (1-year survival: 29.1%) and lowest in those with large cell tumors (1-year survival: 12.8%). Diagnosis in later years, female gender, younger age, either Asian/Pacific Islander or Hispanic race/ethnicity, lower tumor grade, and surgery or beam radiation as part of first-line treatment were generally independently associated with a decreased risk of death, but the prognostic significance of some of these factors (age, ethnicity, tumor grade) varied according to histologic subtype.Conclusion: Findings demonstrate a poor prognosis across histologic subtypes in stage IV NSCLC patients but highlight differences in both absolute survival and the relative importance of select prognostic factors by histologic subclassification. More research using other sources of population-based data could help clarify the role of histology in the presentation, management, and prognosis of late-stage NSCLC.Keywords: epidemiology, nonsmall cell lung cancer, histology, survival

Background: Romosozumab is a novel monoclonal antibody that binds to sclerostin, and has dual effects of increasing bone formation and decreasing bone resorption, giving it a unique mechanism of action. The objective of this study was to perform a systematic review and meta-analysis based on existing worldwide data on treatment effects and safety of romosozumab in randomized controlled trials. Methods: A systematic search was carried out on four databases including PubMed, Embase, Web of Science and Cochrane Central Register of Controlled Trials (CENTRAL). The keywords used for search was “(romosozumab) AND (osteoporosis OR safety)”. Randomized controlled trial or post-hoc studies of the included randomized controlled trial which studied the effects and safety of romosozumab were included. The quality of selected studies was assessed with the Cochrane collaboration tool and the PEDro scale. Results: 20 studies were included for qualitative analysis. 14 studies were included for meta-analysis. In total, there were 13,507 (n = 13,507) participants with 637 men and 12,870 women from original cohorts. The overall mean difference was in favor of romosozumab treatment for lumbar spine (10.04 (95 % confidence interval (CI) = 7.51–12.57; p

At present, new energy is introduced into the power system in large quantities. In order to make it replace the traditional hydropower and gas units as the black start power supply, segmentation restoration strategy of power grid based on the combined optical storage system is studied. Firstly, according to the structure and control mode of the optical storage system, the energy storage system establishes a stable grid-connected bus voltage is proposed, and the photovoltaic system is connected to the grid step by step. Secondly, in order to enhance the damping inertial support of photovoltaic power generation system, virtual synchronous control is proposed to be added to the inverter side of photovoltaic system. Finally, in the process of grid connection between the units to be started and the system load, the energy storage system is configured with additional damping controller to suppress the sub-synchronous oscillation generated by the grid black start, so as to ensure the stable recovery of the grid. In PSCAD/EMTDC, the electromagnetic transient simulation model of black start of the combined optical storage system is built and the simulation analysis is carried out. The results show that the busbar voltage deviation and system frequency deviation in the recovery process of the system are less than 0.65% and less than 1%, which meets the requirements of black start and verifies the feasibility of the start-up strategy proposed in the paper.

Mesembryanthemum crystallinum is a model halophyte that switches from C3 photosynthesis to Crassulacean acid metabolism (CAM) upon extreme abiotic stresses. This study aimed to investigate alkalinity-induced root transcriptome profiling in M. crystallinum. M. crystallinum seedlings were treated with 50 mM sodium bicarbonate (NaHCO3; pH 7.5) and 90 mM NaHCO3 (pH 9.5) for 7 d, respectively. Alkalinity-induced differentially expressed genes (DEGs) were identified and annotated. Functional enrichment analysis was performed for DEGs. The expression of genes related to response to stress and CAM were analyzed and compared. Comparing with control, 50 and 90 mM NaHCO3 treatments induced 4 027 and 25 403 DEGs in M. crystallinum roots, respectively. Among these DEGs, 832 and 131 DEGs were consistently upregulated and downregulated by both stresses, respectively. These genes were associated with multiple biological processes related to response to abiotic stresses. Alkaline stress upregulated genes encoding heat shock proteins and ethylene-related genes, but downregulated genes encoding glutathione S-transferases. Also, genes that encode phosphoenolpyruvate carboxylases, phosphoenolpyruvate carboxylase kinase 1, and malate dehydrogenases related to malate accumulation were upregulated by alkalinity. This study indicated that alkaline stress affected the genes related to stress responses, metabolism, and malate accumulation in the roots of M. crystallinum. [ABSTRACT FROM AUTHOR]

This study evaluates the relationship between the power supply output (voltage (V), current (I), power (W)), used for heating a second dimension (²D) column, and the maximum heating rate and temperature offset achievable by custom built ²D temperature programming system (²DTPS). The influence of the gas chromatograph (GC) oven temperature, ²D column flow rate, and ²D column dimensions when heating the ²D column were also explored. This data provided the minimum power needed for heating a ²D column (normalized to its length), based on the heating rate or temperature offset relative to the first dimension (¹D) column. During the study, optimization of the temperature control algorithm was also carried out to expand the dynamic range of the heating rate for a single power supply output (V, I), while maintaining a temperature deviation of < 0.75 °C between the measured temperature and temperature setpoint. This simplified the power supply specification recommendation to voltages which provide 1.5 A for heating rates and maximum temperature offsets below 28 °C/s and 70 °C, respectively, and 1.9 A when higher heating rates or temperature offsets were needed.

Objective To investigate the effects of PRPF19 knockdown on the proliferation, migration, and invasion of pancreatic cancer cells. Methods The expression of PRPF19 in pancreatic cancer and normal tissues was analyzed using the GEPIA database. The protein and mRNA expression levels of PRPF19 in pancreatic cancer cells were detected by Western blot and qRT-PCR. Small interfering RNA (siRNA) was used to silence the expression of PRPF19 in pancreatic cancer cells, and the knockdown efficiency was verified by Western blot and qRT-PCR. CCK-8, colony forming, and Transwell assay were used to detect the effects of knockdown of PRPF19 on the proliferation, colony forming, migration, and invasion of pancreatic cancer cells. Results GEPIA analysis showed that PRPF19 was highly expressed in pancreatic cancer tissues compared with normal pancreatic tissues. In comparison with normal pancreatic cells, PRPF19 was highly expressed in various pancreatic cancer cell lines such as MIA PaCa-2 and PANC-1 (P < 0.05). In comparison with the control group, PRPF19 knockdown significantly reduced the proliferation rate, colony forming, cell migration, and invasion of pancreatic cancer cells (P < 0.05). Conclusion PRPF19 knockdown inhibits the proliferation, migration, and invasion of pancreatic cancer cells. PRPF19 may play an important role as an oncogene of pancreatic cancer.

Here the authors shows that G-quadruplexes, non-canonical DNA/RNA structures, can have a direct impact on alternative splicing and that binding of splicing regulators is affected by their presence.

The cochlea is a complex organ comprising diverse cell types with highly specialized morphology and function. Until now, the molecular underpinnings of its specializations have mostly been studied from a transcriptional perspective, but accumulating evidence points to post-transcriptional regulation as a major source of molecular diversity. Alternative splicing is one of the most prevalent and well-characterized post-transcriptional regulatory mechanisms. Many molecules important for hearing, such as cadherin 23 or harmonin, undergo alternative splicing to produce functionally distinct isoforms. Some isoforms are expressed specifically in the cochlea, while some show differential expression across the various cochlear cell types and anatomical regions. Clinical phenotypes that arise from mutations affecting specific splice variants testify to the functional relevance of these isoforms. All these clues point to an essential role for alternative splicing in shaping the unique molecular landscape of the cochlea. Although the regulatory mechanisms controlling alternative splicing in the cochlea are poorly characterized, there are animal models with defective splicing regulators that demonstrate the importance of RNA-binding proteins in maintaining cochlear function and cell survival. Recent technological breakthroughs offer exciting prospects for overcoming some of the long-standing hurdles that have complicated the analysis of alternative splicing in the cochlea. Efforts toward this end will help clarify how the remarkable diversity of the cochlear transcriptome is both established and maintained.

A proof-of-concept second dimension (²D) temperature programming system for comprehensive two-dimensional gas chromatography (GC × GC) with true temperature control is introduced and its performance characterized. The ²D temperature programming system utilized a commercial stainless-steel capillary column that was resistively heated and controlled using an Arduino UNO microcontroller. Temperature measurement was accomplished using a differential thermocouple system attached to the ²D column. Diesel was injected in triplicate to compare the reproducibility of the GC × GC system without additional ²D heating, with ²D temperature programming, and with a constant offset, all accomplished using the ²D temperature programming system. Relative standard deviations for the ²D temperature programming were 0.02%, 0.49% and 1.2% for the first dimension (¹D) retention time, ²D retention time, and peak area, respectively. The estimated ²D peak capacity improved by 48% with ²D temperature programming compared to the separation without additional ²D heating.

Summary: Alternative DNA conformations, termed non-B DNA structures, can affect transcription, but the underlying mechanisms and their functional impact have not been systematically characterized. Here, we used computational genomic analyses coupled with massively parallel reporter assays (MPRAs) to show that certain non-B DNA structures have a substantial effect on gene expression. Genomic analyses found that non-B DNA structures at promoters harbor an excess of germline variants. Analysis of multiple MPRAs, including a promoter library specifically designed to perturb non-B DNA structures, functionally validated that Z-DNA can significantly affect promoter activity. We also observed that biophysical properties of non-B DNA motifs, such as the length of Z-DNA motifs and the orientation of G-quadruplex structures relative to transcriptional direction, have a significant effect on promoter activity. Combined, their higher mutation rate and functional effect on transcription implicate a subset of non-B DNA motifs as major drivers of human gene-expression-associated phenotypes.

OBJECTIVE: MicroRNAs (miRNAs) act as critical regulators of genes expression involved in tumor biological processes. The study aimed to investigate the clinical significance and biological role of miR-124 in gastric cancer (GC). PATIENTS AND METHODS: MiR-124 expression was analyzed from 88 GC tissues and adjacent normal tissues by quantitative Real-time PCR (qRT-PCR). Kaplan-Meier curves and log-rank test was used to evaluate the association between miR-124 and the over survival (OS) time of GC patients. MTT assays and transwell invasion assays were performed to assess cell proliferation and invasion. The relationship between miR-124 and Snail2 expression was analyzed by dual luciferase reporter assay. Western blot analyses were performed to detect the relative protein expression. RESULTS: We found that miR-124 expression was significantly reduced in GC tissue samples when compared to the adjacent normal tissues (p<0.05). Lower miR-124 was found to be associated with tumor size (p=0.001), lymphatic metastasis (p=0.008) and TNM stage (p=0.015). Furthermore, patients who have lower miR-124 predicted poor OS time (p<0.05). Function studies suggested that cell proliferation and invasion ability of GC cells were inhibited by up-regulation of miR-124 expression. Moreover, we demonstrated that Snail2 was a direct target of miR-124. Meanwhile, miR-124 inhibited Epithelial-Mesenchymal Transition (EMT) process by repressing the Snail2 expression in GC cells. CONCLUSIONS: MiR-124 acted as a tumor suppressor in GC and may be a useful target for GC treatment. [ABSTRACT FROM AUTHOR]

Objective: Acromegaly is a rare condition and there is often a long path to diagnosis for many patients. We sought to explore patient’s perceptions a nd understanding of acromegaly, to examine the quality of communication and find gap s in the information provided at diagnosis. Design: A prospective study using qualitative research methodology and grounded theory. A semi-structured interview was conducted with 18 patie nts treated for acromegaly in a single tertiary centre and verbatim transcripts were thematically analysed for overarching themes. Results: Eighteen patients with acromegaly were interviewed. The mean a ge of participants was 52 (range 30–72). Four overarching themes emerged; (1) Patients rely on online resources to understand acromegaly in the time between d iagnosis and tertiary care clinic; (2) There is not enough support available for pati ents; (3) Patients have a basic understanding of acromegaly and associated conditions, bu t the long-term impact is underestimated; and (4) Patients initially felt intimidated by the multidisciplinary team panel, but overall found it useful. Conclusion: Acromegalic patients have a strong need for information at the point of initial diagnosis, in particular online resources and interaction with other experienced patients. Wider dissemination of patient educational resources into prima ry and secondary care settings may improve overall patient satisfaction, treatment ad herence and subsequent health care provider–patient relationships.

Alternative polyadenylation (APA) is a widespread and conserved regulatory mechanism that generates diverse 3′ ends on mRNA. APA patterns are often tissue specific and play an important role in cellular processes such as cell proliferation, differentiation, and response to stress. Many APA sites are found in 3′ UTRs, generating mRNA isoforms with different 3′ UTR contents. These alternate 3′ UTR isoforms can change how the transcript is regulated, affecting its stability and translation. Since the subcellular localization of a transcript is often regulated by 3′ UTR sequences, this implies that APA can also change transcript location. However, this connection between APA and RNA localization has only recently been explored. In this review, we discuss the role of APA in mRNA localization across distinct subcellular compartments. We also discuss current challenges and future advancements that will aid our understanding of how APA affects RNA localization and molecular mechanisms that drive these processes.

OBJECTIVES:. To compare the performance of Sequential Organ Failure Assessment, systemic inflammatory response syndrome, Red Flag Sepsis, and National Institute of Clinical Excellence sepsis risk stratification tools in the identification of patients at greatest risk of mortality from sepsis in nonintensive care environments. DESIGN:. Secondary analysis of three annual 24-hour point-prevalence study periods. SETTING:. The general wards and emergency departments of 14 acute hospitals across Wales. Studies were conducted on the third Wednesday of October in 2017, 2018, and 2019. PATIENTS:. We screened all patients presenting to the emergency department and on the general wards. MEASUREMENTS AND MAIN RESULTS:. We recruited 1,271 patients, of which 724 (56.9%) had systemic inflammatory response syndrome greater than or equal to 2, 679 (53.4%) had Sequential Organ Failure Assessment greater than or equal to 2, and 977 (76.9%) had Red Flag Sepsis. When stratified according to National Institute of Clinical Excellence guidelines, 450 patients (35.4%) were in the “High risk” category in comparison with 665 (52.3%) in “Moderate to High risk” and 156 (12.3%) in “Low risk” category. In a planned sensitivity analysis, we found that none of the tools accurately predicted mortality at 90 days, and Sequential Organ Failure Assessment and National Institute of Clinical Excellence tools showed only moderate discriminatory power for mortality at 7 and 14 days. Furthermore, we could not find any significant correlation with any of the tools at any of the mortality time points. CONCLUSIONS:. Our data suggest that the sepsis risk stratification tools currently utilized in emergency departments and on the general wards do not predict mortality adequately. This is illustrated by the disparity in mortality risk of the populations captured by each instrument, as well as the weak concordance between them. We propose that future studies on the development of sepsis identification tools should focus on identifying predicator values of both the short- and long-term outcomes of sepsis.