The Impact of SARS-CoV-2 Infection on the Length of Stay in the Neuro-ICU:A Prospective Multicenter Cohort Study in Eight Neuro-ICU, China Between February and April 2023.

Purpose: The SARS-CoV-2 infection cases are increasing rapidly in neuro-intensive care units (neuro-ICUs) at the beginning of 2023 in China. We aimed to characterize the prevalence, risk factors, and prognosis of critically ill patients treated in neuro-ICUs. Materials and Methods: In the prospective, multicenter, observational registry study, critically ill patients with intracerebral hemorrhage (ICH), subarachnoid hemorrhage (SAH), and traumatic brain injury (TBI) admitted to eight Chinese neuro-ICUs between Feb 16, 2023, to Apr 30, 2023 were enrolled for the study. Mortality and ICU stay day were used as the primary outcomes. Results: 131 patients were finally included and analyzed (mean age 60.36 years [SD 13.81], 64.12% male, 39.69% SARS-CoV-2 infected). The mortality is higher in the SARS-CoV-2 infection group without statistical signification (7.69% vs 5.06%, p>0.05). The length of stay (LOS) in neuro-ICUs was significantly longer among the SARS-CoV-2 infection patients (7(1-12) vs 4(1-8), p<0.01), with increased viral pneumonia occurrence (58.54% vs 7.32%, p<0.01). SARS-CoV-2 infection, surgery, and low GCS scores were independent risk factors for prolonged LOS, and respiratory/renal failure were independent risk factors for death. Conclusion: Based on the present neuro-ICU cohort, SARS-CoV-2 infection was a significant risk for the prolonged LOS of neuro-critically ill patients. Trial Registration: Registered with Chictr.org.cn (ChiCTR2300068355) at 16 February 2023, Prospective registration. https://www.chictr.org.cn/showproj.html?proj=188252.

Estimating the Heavy Metal Contents in Entisols from a Mining Area Based on Improved Spectral Indices and Catboost.

In the study of the inversion of soil multi-species heavy metal element concentrations using hyperspectral techniques, the selection of feature bands is very important. However, interactions among soil elements can lead to redundancy and instability of spectral features. In this study, heavy metal elements (Pb, Zn, Mn, and As) in entisols around a mining area in Harbin, Heilongjiang Province, China, were studied. To optimise the combination of spectral indices and their weights, radar plots of characteristic-band Pearson coefficients (RCBP) were used to screen three-band spectral index combinations of Pb, Zn, Mn, and As elements, while the Catboost algorithm was used to invert the concentrations of each element. The correlations of Fe with the four heavy metals were analysed from both concentration and characteristic band perspectives, while the effect of spectral inversion was further evaluated via spatial analysis. It was found that the regression model for the inversion of the Zn elemental concentration based on the optimised spectral index combinations had the best fit, with R2 = 0.8786 for the test set, followed by Mn (R2 = 0.8576), As (R2 = 0.7916), and Pb (R2 = 0.6022). As far as the characteristic bands are concerned, the best correlations of Fe with the Pb, Zn, Mn and As elements were 0.837, 0.711, 0.542 and 0.303, respectively. The spatial distribution and correlation of the spectral inversion concentrations of the As and Mn elements with the measured concentrations were consistent, and there were some differences in the results for Zn and Pb. Therefore, hyperspectral techniques and analysis of Fe elements have potential applications in the inversion of entisols heavy metal concentrations and can improve the quality monitoring efficiency of these soils.

Visualizing single-molecule conformational transition and binding dynamics of intrinsically disordered proteins.

Intrinsically disordered proteins (IDPs) play crucial roles in cellular processes and hold promise as drug targets. However, the dynamic nature of IDPs remains poorly understood. Here, we construct a single-molecule electrical nanocircuit based on silicon nanowire field-effect transistors (SiNW-FETs) and functionalize it with an individual disordered c-Myc bHLH-LZ domain to enable label-free, in situ, and long-term measurements at the single-molecule level. We use the device to study c-Myc interaction with Max and/or small molecule inhibitors. We observe the self-folding/unfolding process of c-Myc and reveal its interaction mechanism with Max and inhibitors through ultrasensitive real-time monitoring. We capture a relatively stable encounter intermediate ensemble of c-Myc during its transition from the unbound state to the fully folded state. The c-Myc/Max and c-Myc/inhibitor dissociation constants derived are consistent with other ensemble experiments. These proof-of-concept results provide an understanding of the IDP-binding/folding mechanism and represent a promising nanotechnology for IDP conformation/interaction studies and drug discovery.

Graphene-molecule-graphene single-molecule junctions to detect electronic reactions at the molecular scale.

The ability to measure the behavior of a single molecule during a reaction implies the detection of inherent dynamic and static disordered states, which may not be represented when measuring ensemble averages. Here, we describe the building of devices with graphene-molecule-graphene single-molecule junctions integrated into an electrical circuit. These devices are simple to build and are stable, showing tolerance to mechanical changes, solution environment and voltage stimulation. The design of a conductive channel based on a single molecule enables single-molecule detection and is sensitive to variations in physical properties and chemical structures of the detected molecules. The on-chip setup of single-molecule junctions further offers complementary metal-oxide-semiconductor (CMOS) compatibility, enabling logic functions in circuit elements, as well as deciphering of reaction intermediates. We detail the experimental procedure to prepare graphene transistor arrays as a basis for single-molecule junctions and the preparation of nanogapped carboxyl-terminal graphene electrodes by using electron-beam lithography and oxygen plasma etching. We describe the basic design of a molecular bridge with desired functions and terminals to form covalent bonds with electrode arrays, via a chemical reaction, to construct stably integrated single-molecule devices with a yield of 30-50% per chip. The immobilization of the single molecules is then characterized by using inelastic electron tunneling spectra, single-molecule imaging and fluorescent spectra. The whole protocol can be implemented within 2 weeks and requires users trained in using ultra-clean laboratory facilities and the aforementioned instrumentation.

Single-exonuclease nanocircuits reveal the RNA degradation dynamics of PNPase and demonstrate potential for RNA sequencing.

The degradation process of RNA is decisive in guaranteeing high-fidelity translation of genetic information in living organisms. However, visualizing the single-base degradation process in real time and deciphering the degradation mechanism at the single-enzyme level remain formidable challenges. Here, we present a reliable in-situ single-PNPase-molecule dynamic electrical detector based on silicon nanowire field-effect transistors with ultra-high temporal resolution. These devices are capable of realizing real-time and label-free monitoring of RNA analog degradation with single-base resolution, including RNA analog binding, single-nucleotide hydrolysis, and single-base movement. We discover a binding event of the enzyme (near the active site) with the nucleoside, offering a further understanding of the RNA degradation mechanism. Relying on systematic analyses of independent reads, approximately 80% accuracy in RNA nucleoside sequencing is achieved in a single testing process. This proof-of-concept sets up a Complementary Metal Oxide Semiconductor (CMOS)-compatible playground for the development of high-throughput detection technologies toward mechanistic exploration and single-molecule sequencing.

Attenuated effective connectivity of large-scale brain networks in children with autism spectrum disorders.

Introduction: Understanding the neurological basis of autism spectrum disorder (ASD) is important for the diagnosis and treatment of this mental disorder. Emerging evidence has suggested aberrant functional connectivity of large-scale brain networks in individuals with ASD. However, whether the effective connectivity which measures the causal interactions of these networks is also impaired in these patients remains unclear. Objects: The main purpose of this study was to investigate the effective connectivity of large-scale brain networks in patients with ASD during resting state. Materials and methods: The subjects were 42 autistic children and 127 age-matched normal children from the ABIDE II dataset. We investigated effective connectivity of 7 large-scale brain networks including visual network (VN), default mode network (DMN), cerebellum, sensorimotor network (SMN), auditory network (AN), salience network (SN), frontoparietal network (FPN), with spectral dynamic causality model (spDCM). Parametric empirical Bayesian (PEB) was used to perform second-level group analysis and furnished group commonalities and differences in effective connectivity. Furthermore, we analyzed the correlation between the strength of effective connectivity and patients' clinical characteristics. Results: For both groups, SMN acted like a hub network which demonstrated dense effective connectivity with other large-scale brain network. We also observed significant causal interactions within the "triple networks" system, including DMN, SN and FPN. Compared with healthy controls, children with ASD showed decreased effective connectivity among some large-scale brain networks. These brain networks included VN, DMN, cerebellum, SMN, and FPN. In addition, we also found significant negative correlation between the strength of the effective connectivity from right angular gyrus (ANG_R) of DMN to left precentral gyrus (PreCG_L) of SMN and ADOS-G or ADOS-2 module 4 stereotyped behaviors and restricted interest total (ADOS_G_STEREO_BEHAV) scores. Conclusion: Our research provides new evidence for the pathogenesis of children with ASD from the perspective of effective connections within and between large-scale brain networks. The attenuated effective connectivity of brain networks may be a clinical neurobiological feature of ASD. Changes in effective connectivity of brain network in children with ASD may provide useful information for the diagnosis and treatment of the disease.

Transmembrane protein rotaxanes reveal kinetic traps in the refolding of translocated substrates.

Understanding protein folding under conditions similar to those found in vivo remains challenging. Folding occurs mainly vectorially as a polypeptide emerges from the ribosome or from a membrane translocon. Protein folding during membrane translocation is particularly difficult to study. Here, we describe a single-molecule method to characterize the folded state of individual proteins after membrane translocation, by monitoring the ionic current passing through the pore. We tag both N and C termini of a model protein, thioredoxin, with biotinylated oligonucleotides. Under an electric potential, one of the oligonucleotides is pulled through a α-hemolysin nanopore driving the unfolding and translocation of the protein. We trap the protein in the nanopore as a rotaxane-like complex using streptavidin stoppers. The protein is subjected to cycles of unfolding-translocation-refolding switching the voltage polarity. We find that the refolding pathway after translocation is slower than in bulk solution due to the existence of kinetic traps.

Seeking for potential pathogenic genes of major depressive disorder in the Gene Expression Omnibus database.

INTRODUCTION: Major depressive disorder (MDD) is one of the most common mental disorders worldwide. The aim of this study was to identify potential pathological genes in MDD. METHODS: We searched and downloaded gene expression data from the Gene Expression Omnibus database to identify differentially expressed genes (DEGs) in MDD. Then, Kyoto Encyclopedia of Genes and Genomes pathway, Gene Ontology analysis, and protein-protein interaction (PPI) network were applied to investigate the biological function of identified DEGs. The quantitative real-time polymerase chain reaction and a published dataset were used to validate the result of bioinformatics analysis. RESULTS: A total of 514 DEGs were identified in MDD. In the PPI network, some hub genes with high degrees were identified, such as EEF2, RPL26L1, RPLP0, PRPF8, LSM3, DHX9, RSRC1, and AP2B1. The result of in vitro validation of RPL26L1, RSRC1, TOMM20L, RPLPO, PRPF8, AP2B1, STIP1, and C5orf45 was consistent with the bioinformatics analysis. Electronic validation of C5orf45, STIP1, PRPF8, AP2B1, and SLC35E1 was consistent with the bioinformatics analysis. DISCUSSION: The deregulated genes could be used as potential pathological factors of MDD. In addition, EEF2, RPL26L1, RPLP0, PRPF8, LSM3, DHX9, RSRC1, and AP2B1 might be therapeutic targets for MDD.

Isolation and characterization of a heme oxygenase-1 gene from Chinese cabbage.

Heme oxygenase-1 (HO1) is a heme-catabolizing enzyme induced by a variety of stress conditions. This article described the cloning and characterization of BrHO1 gene which codes for a putative HO1 from Chinese cabbage (Brassica rapa subsp. pekinensis). BrHO1 consists of three exons and encodes a protein precursor of 32.3 kD with a putative N-terminal plastid transit peptide. The amino acid sequence of BrHO1 was 84% similar to Arabidopsis counterpart HY1. The three-dimensional structure of BrHO1 showed a high degree of structural conservation compared with the known HO1 crystal structures. Phylogenetic analysis revealed that BrHO1 clearly grouped with the HO1-like sequences. The recombinant BrHO1 protein expressed in Escherichia coli was active in the conversion of heme to biliverdin IXα (BV). Furthermore, the results of subcellular localization of BrHO1 demonstrated that BrHO1 gene product was most likely localized in the chloroplasts. BrHO1 was differently expressed in all tested tissues and could be induced upon osmotic and salinity stresses, cadmium (Cd) exposure, hydrogen peroxide (H(2)O(2)), and hemin treatments. Together, the results suggested that BrHO1 plays an important role in abiotic stress responses.

Cloning and characterization of a heme oxygenase-2 gene from alfalfa (Medicago sativa L.).

Heme oxygenase (HO, EC 1.14.99.3) catalyzes the oxidation of heme and performs vital roles in plant development and stress responses. Two HO isozymes exist in plants. Between these, HO-1 is an oxidative stress-response protein, and HO-2 usually exhibited constitutive expression. Although alfalfa HO-1 gene (MsHO1) has been investigated previously, HO2 is still poorly understood. In this study, we report the cloning and characterization of HO2 gene, MsHO2, from alfalfa (Medica sativa L.). The full-length cDNA of MsHO2 contains an ORF of 870 bp and encodes for 290 amino acid residues with a predicted molecular mass of 33.3 kDa. Similar to MsHO1, MsHO2 also appears to have an N-terminal transit peptide sequence for chloroplast import. Many conserved residues in plant HO were also conserved in MsHO2. However, unlike HO-1, the conserved histidine (His) required for heme-iron binding and HO activity was replaced by tyrosine (Tyr) in MsHO2. Further biochemical activity analysis of purified mature MsHO2 showed no HO activity, suggesting that MsHO2 may not be a true HO in nature. Semi-quantitative RT-PCR confirmed its maximum expression in the germinating seeds. Importantly, the expression levels of MsHO2 were up-regulated under sodium nitroprusside (SNP) and H(2)O(2) (especially) treatment, respectively.

Enhancement of salinity tolerance during rice seed germination by presoaking with hemoglobin.

Salinity stress is an important environmental constraint limiting the productivity of many crops worldwide. In this report, experiments were conducted to investigate the effects of seed presoaking by bovine hemoglobin, an inducer of heme oxygenase-1 (HO-1), on salinity tolerance in rice (Oryza sativa) plants. The results showed that different concentrations of the hemoglobin (0.01, 0.05, 0.2, 1.0, and 5.0 g/L) differentially alleviated the inhibition of rice seed germination and thereafter seedling shoot growth caused by 100 mM NaCl stress, and the responses of 1.0 g/L hemoglobin was the most obvious. Further analyses showed that application of hemoglobin not only increased the HO-1 gene expression, but also differentially induced catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD) activities or transcripts, thus decreasing the lipid peroxidation in germinating rice seeds subjected to salt stress. Compared with non-hemoglobin treatment, hemoglobin presoaking also increased the potassium (K) to sodium (Na) ratio both in the root and shoot parts after salinity stress. The effect is specific for HO-1 since the potent HO-1 inhibitor zinc protoporphyrin IX (ZnPPIX) blocked the positive actions of hemoglobin on seed germination and seedling shoot growth. Overall, these results suggested that hemoglobin performs an advantageous role in enhancement of salinity tolerance during rice seed germination.

[Expression of cyclin A in childhood acute leukemia and its significance].

OBJECTIVE: To investigate the expression of cyclin A protein in childhood acute leukemia (AL) and its significance. METHODS: By using Western blotting analysis, cyclin A protein in bone marrow mononuclear cells from 47 newly diagnosed AL children and 33 non-hematological malignancy children was detected. RESULTS: The expression of cyclin A in AL group (0.38 +/- 0.20) was higher than that in control group (0.03 +/- 0.15) (P < 0.01). The expression of cyclin A in high risk acute lymphocyte leukemia (ALL) group (HR-ALL) (0.62 +/- 0.38) was higher than that in standard risk ALL group (SR-ALL) (0.33 +/- 0.33) (P < 0.05). The expression of cyclin A in WBC > or = 50 x 10(9)/L group and in WBC < 50 x 10(9)/L group was (0.64 +/- 0.36) and (0.39 +/- 0.38), respectively (P < 0.05). Eight (44.4%) out of 18 patients with positive cyclin A expression achieved complete remission (CR). The CR rate was lower than that of patients with negative cyclin A expression (100%) (P < 0.01). CONCLUSIONS: The higher expression of cyclin A may predict a poor prognosis for childhood ALL.

Role of stem cell factor and its receptor in the pathogenesis of pediatric aplastic anemia.

In order to investigate the levels of stem cell factor (SCF) and its receptor c-kit protein and mRNA in pediatric aplastic anemia (AA) and their relevance to the pathogenesis, immunocytochemical and in situ hybridization were utilized to detect the expression of SCF and its receptor c-kit gene protein and mRNA, respectively in 59 children with AA and 51 normal controls. The relationship between SCF and c-kit and the pathogenesis of AA was analyzed subsequently. The results showed that the positive rate of SCF protein and mRNA expression in children with AA was significantly lower than that in healthy controls (P < 0.05). However, there was no significant difference in the positive rate of c-kit protein and mRNA expression between children with AA and control group (P > 0.05). It was concluded that the expression of SCF is significantly decreased in children with AA, which may be closely associated with the pathogenesis of the AA. c-kit may be unrelated to the development of pediatric AA. Therefore, AA in children may have abnormalities at SCF/c-kit signal transduction levels.

Genetic polymorphisms analysis of glutathione S-transferase M1 and T1 in children with acute lymphoblastic leukemia.

The relationship between glutathione S-transferases (GSTs) M1, T1 genotype and childhood acute lymphoblastic leukemia (ALL) was investigated. GSTM1 and GSTT1 genotypes in genomic DNA from 67 children with ALL and 146 healthy controls were analyzed by using the multiplex polymerase chain reaction (PCR). The frequencies of GSTM1, M1-T1 null genotypes in ALL children were significantly higher than in the healthy controls (76.12% versus 52.74%, OR=2.856, P<0.001; 50.74% versus 24.66%, OR=3.148, P<0.001, respectively). However, there was no significant relationship between GSTT1 null genotype and ALL of children (61.19% versus 49.32%, OR=1.621, P>0.05). It was suggested that GSTM1 null genotype might be a risk genotype of childhood ALL, while there as no correlation between GSTT1 null genotype and childhood ALL.

Distribution of variant genotypes of Fc gamma receptor IIIa in healthy Chinese population of Zhengzhou City.

To investigate the distribution of variant genotypes of Fc gamma receptor IIIa (Fc gamma R IIIa) in healthy Chinese population of Zhengzhou city, genomic DNA was extracted from peripheral blood of healthy donators. The genotypes of Fc gamma R IIIa-158 were determined by nested polymerase chain reaction (PCR) in 137 healthy people in Zhengzhou city. The results showed that frequencies of variant genotypes FF, VV and VF were 42.3%, 48.9% and 8.8% respectively. The distribution of Fc gamma R IIIa-158 in healthy Chinese population of Zhengzhou city was polymorphic and different from that of African Americans (AA) and Caucasian Americans (CA).

[Experimental study of DNA synthesis in the airway cells and airway remodeling in asthmatic rats].

OBJECTIVE: To study the DNA synthesis in the airway cells of asthmatic rats after allergen stimulation in association with airway remodeling. METHODS: Double staining immunohistochemical techniques was used to determine DNA synthesis of the airway cells of 12 asthmatic and 12 normal rats. BrdU incorporation into the airway smooth muscle (ASM) and epithelium was quantified by employment of computer-assisted image analysis. RESULTS: BrdU indices in both the ASM and the epithelium of asthmatic model group were higher than those of the control group (P<0.01, P<0.05), and positive linear correlation of the BrdU indices in the ASM and epithelium with the airway diameter was observed (r=0.7828, P<0.01; r=0.5852, P<0.05), which was not found in the control group (r=-0.3755, P>0.05; r=-0.5208, P>0.05). The epithelial thickness of the model group was significantly greater than that of the control group (P<0.01). There was no significant difference in terms of airway diameter, thickness of the ASM and the area positive of alpha-smooth muscle actin between the 2 groups (P>0.05). CONCLUSION: Increased DNA synthesis and accelerated proliferation of ASM and epithelial cells in sensitized SD rats following repeated allergen challenges may lead to airway remodeling.