The number and position of unsaturated bonds in aliphatic aldehydes affect the cysteine-glucose Maillard reaction: Formation mechanism and comparison of volatile compounds.

Nonanal, (E)-2-nonenal, (E,E)-2,4-nonadienal, and (E,Z)-2,6-nonadienal were used to reveal the effect of the number and position of unsaturated bond in aliphatic aldehydes on Maillard reaction for the generation of 88 stewed meat-like volatile compounds. The results showed that (E,E)-2,4-nonadienal and (E,Z)-2,6-nonadienal exhibited greater inhibition of the cysteine reaction with glucose than nonanal and (E)-2-nonenal. However, the positions of the unsaturated bonds in aliphatic aldehydes in the Maillard reaction stage were similar. A carbohydrate module labeling approach was used to present the formation pathways of 34 volatile compounds derived from the Maillard reaction with aliphatic aldehyde systems. The number and position of unsaturated bonds in aliphatic aldehydes generate multiple pathways of flavor compound formation. 2-Propylfuran and (E)-2-(2-pentenyl)furan resulted from aliphatic aldehydes. 5-Butyldihydro-2(3H)-furanone and 2-methylthiophene were produced from the Maillard reaction. 2-Furanmethanol, 2-thiophenecarboxaldehyde, and 5-methyl-2-thiophenecarboxaldehyde were derived from the interaction of aliphatic aldehydes and the Maillard reaction. In Particular, the addition of aliphatic aldehydes changed the formation pathway of 2-propylthiophene, thieno[3,2-b]thiophene, and 2,5-thiophenedicarboxaldehyde. Heatmap and PLS-DA analysis could discriminate volatile compound compositions of the five systems and screen the marker compounds differentiating volatile compounds.

Shaping the Trans-Scale Properties of Schizophrenia via Cerebral Alterations on Magnetic Resonance Imaging and Single-Nucleotide Polymorphisms of Coding and Non-Coding Regions.

Schizophrenia is a complex mental illness with genetic heterogeneity, which is often accompanied by alterations in brain structure and function. The neurobiological mechanism of schizophrenia associated with heredity remains unknown. Recently, the development of trans-scale and multi-omics methods that integrate gene and imaging information sheds new light on the nature of schizophrenia. In this article, we summarized the results of brain structural and functional changes related to the specific single-nucleotide polymorphisms (SNPs) in the past decade, and the SNPs were divided into non-coding regions and coding regions, respectively. It is hoped that the relationship between SNPs and cerebral alterations can be displayed more clearly and intuitively, so as to provide fresh approaches for the discovery of potential biomarkers and the development of clinical accurate individualized treatment decision-making.

A proteomic platform to identify off-target proteins associated with therapeutic modalities that induce protein degradation or gene silencing.

Novel modalities such as PROTAC and RNAi have the ability to inadvertently alter the abundance of endogenous proteins. Currently available in vitro secondary pharmacology assays, which evaluate off-target binding or activity of small molecules, do not fully assess the off-target effects of PROTAC and are not applicable to RNAi. To address this gap, we developed a proteomics-based platform to comprehensively evaluate the abundance of off-target proteins. First, we selected off-target proteins using genetics and pharmacology evidence. This process yielded 2813 proteins, which we refer to as the "selected off-target proteome" (SOTP). An iterative algorithm was then used to identify four human cell lines out of 932. The 4 cell lines collectively expressed ~ 80% of the SOTP based on transcriptome data. Second, we used mass spectrometry to quantify the intracellular and extracellular proteins from the selected cell lines. Among over 10,000 quantifiable proteins identified, 1828 were part of the predefined SOTP. The SOTP was designed to be easily modified or expanded, owing to the rational selection process developed and the label free LC-MS/MS approach chosen. This versatility inherent to our platform is essential to design fit-for-purpose studies that can address the dynamic questions faced in investigative toxicology.

Altered functional connectivity of the dentate nuclei in patients with schizophrenia.

Dentate nuclei (DN) are vital structures in the anatomical circuits that link the cerebellum to the cerebrum. However, the characteristics of DN functional connectivity (FC) in schizophrenia remain largely unknown. In this study, we investigated the FC of the DN in patients with schizophrenia and examined their possible clinical correlates using resting-state functional magnetic imaging data. We found that the patient group had greater DN FC with the parietal lobe (e.g., postcentral gyrus and superior parietal lobule) and less DN FC with the prefrontal cortex (e.g., superior frontal gyrus), posterior cingulate cortex, and regional cerebellum (e.g., vermis 4-5 and crus I) than did the control group. Furthermore, some abnormal connectivities of the DN with these regions significantly correlated with psychiatric symptoms. These results suggest that the DN circuits are disturbed and may participate in the pathophysiology of schizophrenia.

Nonclinical Safety Assessment of AMG 553, an Investigational Chimeric Antigen Receptor T-Cell Therapy for the Treatment of Acute Myeloid Leukemia.

Feline McDonough Sarcoma-like tyrosine kinase 3 (FLT3), a tyrosine-protein kinase involved in hematopoiesis, is detectable on the cell surface of approximately 80% of leukemia isolates from adult patients with acute myeloid leukemia (AML). AMG 553 is an investigational chimeric antigen receptor (CAR) T-cell immunotherapy for the treatment of AML. FLT3 expression analysis and in vitro and in vivo studies were leveraged to evaluate the nonclinical safety of AMG 553. Cynomolgus monkeys administered autologous anti-FLT3 CAR T cells demonstrated no evidence of CAR T-cell-mediated toxicity, expansion, or persistence, likely due to restricted cell surface FLT3 protein expression in healthy animals. This highlights the limited value of such in vivo studies for safety assessment of the CAR T-cell modality when directed against a target with restricted expression. To complement these studies and directly evaluate the potential toxicities of eliciting T-cell-mediated cytotoxicity against cells with surface expression of FLT3 protein in vivo, data from cynomolgus monkey toxicology studies with 2 bispecific T-cell engager molecules targeting FLT3 were leveraged; findings were consistent with the targeted killing of bone marrow cells expressing cell surface FLT3. Potential AMG 553-induced cytotoxicity was assessed against a wide range of normal human primary cells and cell lines; cytotoxicity was observed against FLT3-positive AML cell lines and a percentage of primary bone marrow CD34+ cells. In conclusion, the nonclinical safety data suggest that AMG 553 can target FLT3 protein on AML cells, whereas only affecting a percentage of normal hematopoietic stem and progenitor cells, supporting clinical development.

[Effect of high-flow nasal cannula oxygen therapy on diaphragmatic function in patients with acute exacerbation of chronic obstructive pulmonary disease: a prospective randomized controlled trial].

OBJECTIVE: To investigate the effect of high-flow nasal cannula oxygen therapy (HFNC) on the clinical efficacy and diaphragm function of patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: The patients with mild to moderate AECOPD (clinical classification I-II) admitted to Huxi Hospital Affiliated to Jining Medical College from January to October in 2018 were enrolled. The patients were divided into HFNC treatment group and routine oxygen therapy control group (each n = 37) by randomly number table method. The two groups were given bronchiectasis drugs, corticosteroids, expectorant, anti-infection treatment, at the same time, the HFNC treatment group was given HFNC with the initial flow rate of 40 L/min. The routine oxygen therapy control group was given low flow oxygen, and the initial flow rate was 3 L/min. General data such as gender, age, clinical grade, acute physiology and chronic health evaluation II (APACHE II) score were recorded. Bedside ultrasound was used to measure the diaphragmatic excursions during quiet breathing (DEq), diaphragmatic excursions during deep breathing (DEd), and diaphragmatic shallow fast breathing index (D-RSBI) before and 2, 24 and 48 hours after treatment in both groups and compared, meanwhile, arterial blood gas analysis was performed, and arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2) were recorded. RESULTS: Two patients in the HFNC treatment group withdrew from the study because they could not tolerate HFNC, while other patients were enrolled in the analysis. There was no statistically significant difference in gender, age, proportion of AECOPD II grade or APACHE II score between the two groups, indicating that the general data of the two groups were comparable and balanced. There was no statistically significant difference in DEq, DEd, D-RSBI, PaO2 or PaCO2 before treatment between the two groups. After treatment, DEp in both groups was decreased gradually with time, it was decreased earlier in the HFNC treatment group, and it showed significant difference as compared with that before treatment at 2 hours after treatment (mm: 18.3±3.1 vs. 20.1±4.2, P < 0.01), and it was significantly lower than that in the routine oxygen therapy control group (mm: 18.3±3.1 vs. 20.3±3.7, P < 0.05); DEd was gradually increased in both groups, it was significantly increased in the HFNC treatment group, and it was significantly higher than that in the routine oxygen therapy control group at 24 hours and 48 hours after treatment (mm: 55.2±7.6 vs. 50.8±9.2 at 24 hours, 59.4±7.7 vs. 53.6±9.1 at 48 hours, both P < 0.05); D-RSBI was decreased gradually in both groups, it was decreased earlier and more significant in the HFNC treatment group, and it was significantly lower than that in routine oxygen therapy control group at 24 hours and 48 hours after treatment (times×min-1×mm-1: 0.41±0.13 vs. 0.51±0.20 at 24 hours, 0.31±0.12 vs. 0.43±0.17 at 48 hours, both P < 0.05). After treatment, there was no statistically significant difference in PaO2 or PaCO2 between the two groups. CONCLUSIONS: HFNC can effectively relieve diaphragm fatigue in patients with mild to moderate AECOPD, but it had no effect on carbon dioxide retention.

Precise temporal regulation of alternative splicing during neural development.

Alternative splicing (AS) is one crucial step of gene expression that must be tightly regulated during neurodevelopment. However, the precise timing of developmental splicing switches and the underlying regulatory mechanisms are poorly understood. Here we systematically analyze the temporal regulation of AS in a large number of transcriptome profiles of developing mouse cortices, in vivo purified neuronal subtypes, and neurons differentiated in vitro. Our analysis reveals early-switch and late-switch exons in genes with distinct functions, and these switches accurately define neuronal maturation stages. Integrative modeling suggests that these switches are under direct and combinatorial regulation by distinct sets of neuronal RNA-binding proteins including Nova, Rbfox, Mbnl, and Ptbp. Surprisingly, various neuronal subtypes in the sensory systems lack Nova and/or Rbfox expression. These neurons retain the "immature" splicing program in early-switch exons, affecting numerous synaptic genes. These results provide new insights into the organization and regulation of the neurodevelopmental transcriptome.

Systematic discovery of regulated and conserved alternative exons in the mammalian brain reveals NMD modulating chromatin regulators.

Alternative splicing (AS) dramatically expands the complexity of the mammalian brain transcriptome, but its atlas remains incomplete. Here we performed deep mRNA sequencing of mouse cortex to discover and characterize alternative exons with potential functional significance. Our analysis expands the list of AS events over 10-fold compared with previous annotations, demonstrating that 72% of multiexon genes express multiple splice variants in this single tissue. To evaluate functionality of the newly discovered AS events, we conducted comprehensive analyses on central nervous system (CNS) cell type-specific splicing, targets of tissue- or cell type-specific RNA binding proteins (RBPs), evolutionary selection pressure, and coupling of AS with nonsense-mediated decay (AS-NMD). We show that newly discovered events account for 23-42% of all cassette exons under tissue- or cell type-specific regulation. Furthermore, over 7,000 cassette exons are under evolutionary selection for regulated AS in mammals, 70% of which are new. Among these are 3,058 highly conserved cassette exons, including 1,014 NMD exons that may function directly to control gene expression levels. These NMD exons are particularly enriched in RBPs including splicing factors and interestingly also regulators for other steps of RNA metabolism. Unexpectedly, a second group of NMD exons reside in genes encoding chromatin regulators. Although the conservation of NMD exons in RBPs frequently extends into lower vertebrates, NMD exons in chromatin regulators are introduced later into the mammalian lineage, implying the emergence of a novel mechanism coupling AS and epigenetics. Our results highlight previously uncharacterized complexity and evolution in the mammalian brain transcriptome.

Clinical effect analysis of microscopic surgery for epiglottis cysts with coblation.

This study aims to explore the effects and advantages of coblation combined with microscopy to treat epiglottis cysts. Ninety patients with epiglottis cysts were randomly assigned to three groups: the first group: marsupialisation + electric coagulation group, n = 30; the second group: marsupialisation + coblation, n = 30; and the third group: marsupialisation + coblation + microsurgery, n = 30. To compare the cure rate, intraoperative bleeding volume, postoperative pain, operation time and postoperative complications were investigated among these three groups. The comparison among three procedures showed a significant difference for intraoperative bleeding volume, operation time and postoperative pain (P < 0.05), whereas no significant difference was observed for cure rate (P > 0.05). These three procedures are effective in treating epiglottis cysts. Microscopic surgery with coblation has the advantages of less bleeding, short procedure duration, less pain and few complications. Thus, microscopic surgery is worthy of clinical application.

Experimental studies on the inhibition of adenovirus-ING4-OSM therapy on nasopharyngeal carcinoma proliferation in vitro and in vivo.

In the present study, the effects of the co-transfer of the tumor growth inhibitor 4 gene (ING4) together with the Oncostatin M (OSM) were investigated on tumor regression and subsequent tumor recurrence. We constructed a recombinant adenovirus carrying ING4 and OSM, which could induce high-level expression of these three genes in NPC CNE-1 cells. Ad-ING4, Ad-OSM and Ad-ING4-OSM infection all inhibited the growth of CNE-1 cells in vitro, while the Ad-ING4-OSM exerted the strongest inhibitory effect. In CNE-1 xenograft tumor models mice, an intratumoral injection of Ad-ING4, Ad-OSM and Ad-ING4-OSM resulted in a reduced tumor burden, compared to normal saline controls. Therefore, we suggested that the introduction of adenovirus-mediated ING4 and OSM genes could synergistically decrease the recurrence or metastases and develop a control of NPC tumors, which advocate a promising therapeutic future in NPC treatment.

HITS-CLIP and integrative modeling define the Rbfox splicing-regulatory network linked to brain development and autism.

The RNA binding proteins Rbfox1/2/3 regulate alternative splicing in the nervous system, and disruption of Rbfox1 has been implicated in autism. However, comprehensive identification of functional Rbfox targets has been challenging. Here, we perform HITS-CLIP for all three Rbfox family members in order to globally map, at a single-nucleotide resolution, their in vivo RNA interaction sites in the mouse brain. We find that the two guanines in the Rbfox binding motif UGCAUG are critical for protein-RNA interactions and crosslinking. Using integrative modeling, these interaction sites, combined with additional datasets, define 1,059 direct Rbfox target alternative splicing events. Over half of the quantifiable targets show dynamic changes during brain development. Of particular interest are 111 events from 48 candidate autism-susceptibility genes, including syndromic autism genes Shank3, Cacna1c, and Tsc2. Alteration of Rbfox targets in some autistic brains is correlated with downregulation of all three Rbfox proteins, supporting the potential clinical relevance of the splicing-regulatory network.

[Correlation between allergic rhinitis and childhood obstructive sleep apnea-hypopnea syndrome].

OBJECTIVE: To investigate the correlation between allergic rhinitis and obstructive sleep apnea-hypopnea (OSAHS) syndrome in children. METHOD: (1) According to medical history, physical signs, skin-prick test, serum sIgE, endoscopic examination and polysomnography, the incidence of allergic rhinitis was confirmed in 574 cases of childhood obstructive sleeping apnea-hypopnea syndrome in our hospital between July in 2008 to June in 2010. (2) Effects of anti-allergic drugs were observed on 78 children with OSAHS and allergic rhinitis meanwhile. RESULT: (1) 258 cases of allergic rhinitis were confirmed in 574 cases of OSAHS, accounting for 44.9% of the OSAHS cases and 50.4% of all cases of allergic rhinitis during the same period. Most of them were perennial allergic rhinitis (223 cases, 86.4%), and 72.5% of them were aroused by fungal allergen. Compared with other allergen, statistically significant difference was found (P < 0.05). (2) After receiving anti-allergic drugs regularly for 3 weeks, 40 cases suffering from mild and moderate OSAHS and allergic rhinitis, 3 cases out of 38 cases suffering from serious OSAHS and allergic rhinitis showed satisfactory results, while other cases had little improvement. CONCLUSION: Allergic rhinitis is closely related to childhood OSAHS, and perennial allergic rhinitis dominates. The most common allergen is fungal allergen, the second is house and flour dust mites. So for patients of mild and moderate OSAHS with allergic rhinitis, regular anti-allergic drugs can lighten OSAHS effectively and may make patients avoid surgery. Severe OSAHS cases can receive surgical intervention if prior anti allergic therapy fails. Anti allergic therapy should be adopted routinely after tonsillectomy and adenoidectomy in case of hypopnea due to hypertrophy of inferior turbinate or tubal torus in allergic rhinitis.

Evolution of CpG island promoter function underlies changes in KChIP2 potassium channel subunit gene expression in mammalian heart.

Scaling of cardiac electrophysiology with body mass requires large changes in the ventricular action potential duration and heart rate in mammals. These changes in cellular electrophysiological function are produced by systematic and coordinated changes in the expression of multiple ion channel and transporter genes. Expression of one important potassium current, the transient outward current (I(to)), changes significantly during mammalian evolution. Changes in I(to) expression are determined, in part, by variation in the expression of an obligatory auxiliary subunit encoded by the KChIP2 gene. The KChIP2 gene is expressed in both cardiac myocytes and neurons and transcription in both cell types is initiated from the same CpG island promoter. Species-dependent variation of KChIP2 expression in heart is mediated by the evolution of the cis-regulatory function of this gene. Surprisingly, the major locus of evolutionary change for KChIP2 gene expression in heart lies within the CpG island core promoter. The results demonstrate that CpG island promoters are not simply permissive for gene expression but can also contribute to tissue-selective expression and, as such, can function as an important locus for the evolution of cis-regulatory function. More generally, evolution of the cis-regulatory function of voltage-gated ion channel genes appears to be an effective and efficient way to modify channel expression levels to optimize electrophysiological function.

Robust L-type calcium current expression following heterozygous knockout of the Cav1.2 gene in adult mouse heart.

Mechanisms that contribute to maintaining expression of functional ion channels at relatively constant levels following perturbations of channel biosynthesis are likely to contribute significantly to the stability of electrophysiological systems in some pathological conditions. In order to examine the robustness of L-type calcium current expression, the response to changes in Ca²âº channel Cav1.2 gene dosage was studied in adult mice. Using a cardiac-specific inducible Cre recombinase system, Cav1.2 mRNA was reduced to 11 ± 1% of control values in homozygous floxed mice and the mice died rapidly (11.9 ± 3 days) after induction of gene deletion. In these homozygous knockout mice, echocardiographic analysis showed that myocardial contractility was reduced to 14 ± 1% of control values shortly before death. For these mice, no effective compensatory changes in ion channel gene expression were triggered following deletion of both Cav1.2 alleles, despite the dramatic decay in cardiac function. In contrast to the homozygote knockout mice, following knockout of only one Cav1.2 allele, cardiac function remained unchanged, as did survival.Cav1.2mRNAexpression in the left ventricle of heterozygous knockout mice was reduced to 58 ± 3% of control values and there was a 21 ± 2% reduction in Cav1.2 protein expression. There was no significant reduction in L-type Ca²âº current density in these mice. The results are consistent with a model of L-type calcium channel biosynthesis in which there are one or more saturated steps, which act to buffer changes in both total Cav1.2 protein and L-type current expression.

[More exploration on endoscopic rhinoplasty and septoplasty].

OBJECTIVE: Exploring a method of rhinoplasty and septoplasty to get better nasal function and aesthetic effect. METHOD: 1) Using endoscopic rhinoplasty and septoplasty, separating the joint of quadrangular cartilage and the bones around it, resecting vertical and horizontal strip of the deviated septal cartilage and fracturing the deviated septal bones and reconstructing them with three layers (mucosa-cartilage or bone-mucosa), positing the reconstructed septum in the middle between two lateral walls of nasal cavity, proper space between septum and turbinates was maintained. 2) The caudal part of quadrangular cartilage was resected and placed in the columella pocket between the two medial parts of the alar cartilage and the supporting function of the septal cartilage was maintained. RESULT: Compared with traditional nasal septal reconstruction surgery, our method got better functional and aesthetic outcome. CONCLUSION: The anatomic three layers were preserved in our method, thus it is better than the traditional methods. The caudal septal cartilage should be used as the perfect material in rhinoplasty.

[Detection of PD4, CD44, PCNA protein and its clinical significance in human laryngeal carcinoma].

OBJECTIVE: To study the expression of PD4, CD44 and PCNA proteins in laryngeal carcinoma and their relationships with the pathogenesis, development and prognosis of laryngeal carcinoma. METHOD: Immunohistochemistry was used to study 140 cases of laryngeal carcinoma tissues, 25 cases of precarcinoma tissues, 36 cases of vocal cord polyps and 13 cases of normal tissues adjacent to laryngeal carcinoma. RESULT: 1. The positive rates of PD4, CD44 and PCNA were 45.71% (64/140), 64.29% (90/140) and 77.86% (109/140) in laryngeal carcinoma, which were much higher than in non-carcinoma tissues (P < 0.01). 2. The third and fourth stages laryngeal carcinoma express stronger PD4 and CD44 than those of the first and second stages. Laryngeal carcinoma with cervical metastasis had higher expression than those without cervical metastasis. To 3 and 5 years' survival, PD4, CD44 and PCNA positive cases had lower chance than those negative cases(P < 0.01 or 0.05). 3. The over all positive rate of PD4, CD44 and PCNA was 27.86% (39/140) in laryngeal carcinoma tissues and 5.41% (4/74) in non-carcinoma tissues. CONCLUSION: The high expression of PD4, CD44 and PCNA proteins maybe closely related to the pathogenesis, development and prognosis of laryngeal carcinoma.

Evolution of ventricular myocyte electrophysiology.

The relative importance of regulatory versus structural evolution for the evolution of different biological systems is a subject of controversy. The primacy of regulatory evolution in the diversification of morphological traits has been promoted by many evolutionary developmental biologists. For physiological traits, however, the role of regulatory evolution has received less attention or has been considered to be relatively unimportant. To address this issue for electrophysiological systems, we examined the importance of regulatory and structural evolution in the evolution of the electrophysiological function of cardiac myocytes in mammals. In particular, two related phenomena were studied: the change in action potential morphology in small mammals and the scaling of action potential duration across mammalian phylogeny. In general, the functional properties of the ion channels involved in ventricular action potential repolarization were found to be relatively invariant. In contrast, there were large changes in the expression levels of multiple ion channel and transporter genes. For the Kv2.1 and Kv4.2 potassium channel genes, which are primary determinants of the action potential morphology in small mammals, the functional properties of the proximal promoter regions were found to vary in concordance with species-dependent differences in mRNA expression, suggesting that evolution of cis-regulatory elements is the primary determinant of this trait. Scaling of action potential duration was found to be a complex phenomenon, involving changes in the expression of a large number of channels and transporters. In this case, it is concluded that regulatory evolution is the predominant mechanism by which the scaling is achieved.

[Expression of PD4, CD44v6, CD44v9 and its clinical significance in human laryngeal carcinoma].

OBJECTIVE: Study on the role of mycoplasma infection and expression of CD44v6, CD44v9 protein in the pathogenesis, development and prognosis of laryngeal carcinoma (LC). METHOD: Immunohistochemistry was used to study 137 cases of laryngeal carcinoma tissues, 26 cases of precarcinoma tissues, 34 cases of vocal cord polypus and 15 cases of normal tissues adjacent to laryngeal carcinoma. RESULT: The positive rate of PD4 and CD44v6, CD44v9 was 45.3% (62/137), 72.3% (99/137) and 56.2% (77/137) in laryngeal carcinoma, which are much higher than those in non-carcinoma tissues. (2) The positive rate of PD4 and CD44v6. CD44v9 in the third and fourth stages of LC were higher than those in the first and second stages of LC. Laryngeal carcinoma with cervical metastasis had higher expression than those without cervical metastasis (P < 0.01). To 3 and 5 years survival, PD4 and CD44v6, CD44v9 positive cases had lower chance than those negative cases (P < 0.01 [Chinese character: see text] 0.05). (3) The overall positive rate of PD4 and CD44v6, CD44v9 was 27.7% (38/137) in laryngeal carcinoma tissues and 5.3% (4/75) in non-carcinoma tissues. CONCLUSION: These results implicates that the pathogenesis, development and prognosis of laryngeal carcinoma maybe closely related to the high expression of PD4 and CD44v6, CD44v9 protein.