Combined prognostic significance of white blood cell count and D-dimer on in-hospital outcomes of acute ischemic stroke.

BACKGROUND AND AIMS: The prognostic significance of combination of white blood cell (WBC) and D-dimer on acute ischemic stroke (AIS) remains to be explored. We aimed to investigate the combined effect of WBC and D-dimer levels on in-hospital outcomes of AIS patients. METHODS AND RESULTS: 801 AIS patients were included. Patients were divided into four groups according to the cut-point identified by receiver operating characteristic (ROC) curve of D-dimer (1.105 µg/L) and WBC (7.05 × 109/L): LWLD (low WBC count and low D-dimer), LWHD (low WBC count and high D-dimer), HWLD (high WBC count and low D-dimer), and HWHD (high WBC count and high D-dimer). HWHD group had the highest cumulative incidence of in-hospital mortality (hazard ratio, 5.79; 95%CI, 1.71-19.58, P = 0.006). Patients in HWHD group were 4.14 fold more likely to have in-hospital pneumonia (odds ratio, 4.14; 95%CI, 2.09-8.21; P < 0.001), compared with those in LWLD group. The area under curve (AUC) of the combination of WBC and D-dimer levels for in-hospital mortality and pneumonia was larger than that of WBC and D-dimer alone (0.920 vs. 0.900 vs. 0.915; 0.831 vs. 0.829 vs. 0.807). CONCLUSIONS: The combination of WBC count and D-dimer levels at admission was independently associated with in-hospital outcomes of AIS patients. The addition of WBC to D-dimer levels had a tendency to improve the predictive power for in-hospital mortality and pneumonia.

A new clinical score to predict the possibility of stroke patients receiving intravenous thrombolysis.

BACKGROUND: We aimed to develop and validate a clinical score to identify the factors which contribute to variation in, and influence clinician's decision-making about treating acute ischemic stroke (AIS) patients with Intravenous thrombolysis (IVT). METHODS: We retrospectively included consecutive AIS patients within 4.5 hours after onset in the emergency department (ED), who were admitted to a comprehensive stroke center in Jiangsu province, China. The patients were randomly divided into derivation (60%) and validation data sets (40%) to develop and validate the clinical score. Multivariable stepwise forward logistic regression was performed to identify the independent predictors of IVT offering in the derivation data. RESULTS: Out of 526 included patients, 418 patients received thrombolytic therapy. Nine patient factors were associated with the likelihood of thrombolysis (age, time to hospital, National Institute of Health stroke scale (NIHSS) score, great vessel, facial paralysis, dizziness, headache, history of stroke, and neutrophil ratio). The c-statistics of the Intravenous Thrombolysis Score in the derivation cohort (n= 316) and validation cohort(n = 210) were 0.795 and 0.751, respectively. The performance of the scoring model was validated with a calibration plot showing good predictive accuracy for the scores in the derivation data (calibrated P = 0.861) and validation data (calibrated P = 0.876). CONCLUSIONS: The Intravenous Thrombolysis Score for predicting the possibility of offering IVT to AIS patients indicates that clinicians differ in their thresholds for the treatment across a number of patient-related factors, which will be linked to training professional development programmes and address the impact of non-medical influences on decision-making using evidence-based strategies.

Genome-wide identification and functional analysis of circRNAs in Trichophyton rubrum conidial and mycelial stages.

BACKGROUND: Circular RNAs (circRNAs) are a group of noncoding RNAs that participate in gene expression regulation in various pathways. The essential roles of circRNAs have been revealed in many species. However, knowledge of circRNAs in fungi is still not comprehensive. RESULTS: Trichophyton rubrum (T. rubrum) is considered a model organism of human pathogenic filamentous fungi and dermatophytes. In this study, we performed a genome-wide investigation of circRNAs in T. rubrum based on high-throughput sequencing and ultimately identified 4254 circRNAs. Most of these circRNAs were specific to the conidial or mycelial stage, revealing a developmental stage-specific expression pattern. In addition, 940 circRNAs were significantly differentially expressed between the conidial and mycelial stages. PCR experiments conducted on seven randomly selected differentially expressed (DE-) circRNAs confirmed the circularized structures and relative expression levels of these circRNAs. Based on their genome locations, most circRNAs originated from intergenic regions, unlike those in plants and animals. Furthermore, we constructed circRNA-miRNA-mRNA regulatory networks that included 661 DE-circRNAs targeting 140 miRNAs and further regulating 2753 mRNAs. The relative expression levels of two randomly selected circRNA-miRNA-mRNA axes were investigated by qRT-PCR, and the competing endogenous RNA (ceRNA) network theory was validated. Functional enrichment analysis of the target genes suggested that they were significantly involved in posttranscriptional processes and protein synthesis as well as some small-molecule metabolism processes. CircRNAs are relatively more conserved in closely related dermatophytes but rarely conserved in distantly related species. Tru_circ07138_001 is a highly conserved circRNA that was conserved in all ten dermatophytes analyzed in our study and three distantly related species. Its host gene TERG_07138 was also highly conserved in two of these distantly related species Gallus gallus and Caenorhabditis elegans. The specific role of this circRNA deserves further exploration. CONCLUSIONS: Our study is the first to provide a global profile of circRNAs in T. rubrum as well as dermatophytes. These results could serve as valuable resources for research on circRNA regulatory mechanisms in fungi and reveal new insights for further investigation of the physical characteristics of these significant human fungal pathogens.

The amino acid and carnitine concentration changes in bronchoalveolar lavage fluid from lung cancer patients.

OBJECTIVE: To screen out potential biomarkers by analyzing fundamental nutrients in the bronchoalveolar lavage fluid (BALF) before confirming the lung cancer. METHODS: In this study, 44 patients were enrolled with clinical information. The concentrations of 23 amino acids and 35 carnitines in their BALF were detected with the high-performance liquid chromatography-mass spectrometry (HPLC-MS). Combined with clinicopathological diagnosis, the patients were divided into the lung cancer group (grades I & II and III & IV) and the non-cancer group for standard statistical analysis. RESULTS: The partial least squares-discriminant analysis (PLS-DA), the Shapiro-Wilk test, and the Bonferroni correction results showed that the serine concentration was higher and the butane-diacyl-carnitine (C4DC) concentration was lower in the lung cancer group, further showing the same changing trend continuously through the non-cancer stage, grades I & II stage and grades III & IV stage. Those two potential biomarkers have been identified. CONCLUSION: The HPLC-MS target detection in clinic for nutrient concentration levels is a promising technique to find the changing concentration of serine and C4DC in BALF, which provides an economical and practical way for early warning of lung cancer.

Does prolonged emergency department length of stay(EDLOS) affect the outcomes of acute ischemic stroke patients?

BACKGROUND AND PURPOSE: The effect of emergency department length of stay (EDLOS) on outcomes of patients with acute ischemic stroke (AIS) remains largely unexamined. We aimed to investigate the association between EDLOS and outcomes in AIS patients. METHODS: 618 AIS patients were enrolled. Baseline demographics, vascular risk factors, ED admission information, hyperacute treatment of AIS and stroke severity were collected. Stroke progression was defined as any new neurological symptoms/signs or any neurological worsening within 7 days after stroke onset and poor prognosis was defined as modified Rankin Scale(mRS) scores>2 at 30 day. The effect of EDLOS on stroke progression and prognosis was assessed. RESULTS: The median EDLOS was 2.5 h (1.4-6.9 h). On multivariable linear regression, presentation month between Apr. and Jun., admission at the ED between 7 am to 3 pm(P = 0.036), transferring to stroke unit, receiving endovascular interventional treatment, onset on holidays, and progressive stroke were associated with shorter EDLOS(all P < 0.05). A shorter EDLOS was significantly associated with an increased risk of stroke progression (P = 0.007). Patients with the lowest EDLOS (≤1.35 h) were 2-3 fold more likely to have stroke progression, compared with those with the highest EDLOS (>6.93 h) (OR, 2.52; 95% CI, 1.29-4.93; P = 0.043). However, no significant association between EDLOS and stroke prognosis was revealed. CONCLUSIONS: In AIS patients, shorter EDLOS was associated with the increased risk of stroke progression, possibly reflecting prioritized admission of more severely affected patients at high risk of stroke progression. EDLOS alone might be an insufficient indicator of stroke care in the ED.

Polysialic-Acid-Based Micelles Promote Neural Regeneration in Spinal Cord Injury Therapy.

Spinal cord injury (SCI) routinely causes the immediate loss and disruption of neurons followed by complicated secondary injuries, including inflammation, oxidative stress, and dense glial scar formation. Inhibitory factors in the lesion scar and poor intrinsic neural regeneration capacity restrict functional recovery after injury. Minocycline, which has neuroprotective activity, can alleviate secondary injury, but the long-term administration of this drug may cause toxicity. Polysialic acid (PSA) is a large cell-surface carbohydrate that is critical for central nervous system development and is capable of promoting precursor cell migration, axon path finding, and synaptic remodeling; thus, PSA plays a vital role in tissue repair and regeneration. Here, we developed a PSA-based minocycline-loaded nanodrug delivery system (PSM) for the synergistic therapy of spinal cord injury. The prepared PSM exerted marked anti-inflammatory and neuroprotective activities both in vitro and in vivo. The administration of PSM could significantly protect neurons and myelin sheaths from damage, reduce the formation of glial scar, recruit endogenous neural stem cells to the lesion site, and promote the regeneration of neurons and the extension of long axons throughout the glial scar, thereby largely improving the locomotor function of SCI rats and exerting a superior therapeutic effect. The findings might provide a novel strategy for SCI synergistic therapy and the utilization of PSA in other central nervous system diseases.

Factors Associated with Pre-Hospital Delay and Intravenous Thrombolysis in China.

BACKGROUND: Pre-hospital delay was a critical factor affecting stroke patients receiving intravenous thrombolytic therapy. The aim of this study was to explore the factors associated with pre-hospital delay and thrombolysis in China. METHODS: Patient data were obtained from emergency department (ED), and the factors of patient pre-hospital delay were recorded through a well-designed form. RESULTS: A total of 630 patients were eventually included in the study. 317 patients were admitted to the ED during the thrombolysis time window, and only 105 patients received intravenous thrombolytic therapy. In the univariate analysis, transportation (OR: 0.15; 95% CI: 0.44 - 0.518; p = 0.001), atrial fibrillation (OR: 0.555; 95% CI: 0.372-0.828; p = 0.004) and response of symptoms (OR: 0.002; 95% CI: 0.000-0.013; p = 0.000) were associated with early arrival. Speech disturbances (OR: 2.095; 95% CI: 1.294-3.391; p = 0.002), smoking (OR: 2.563; 95% CI: 1.527-4.304; p = 0.000), alcohol consumption (OR: 2.155; 95% CI: 1.159-4.005; p = 0.014) and referral presentation (OR: 2.837; 95% CI: 1.584-5.082; p = 0.000) were associated with thrombolysis. In the logistic regression analysis, direct visiting to the hospital after onset and rushing to emergency after onset were independent predictor of early arrival of AIS and intravenous thrombolytic. CONCLUSIONS: The pre-hospital delay of acute ischemic stroke in China was still serious. Strengthening the ability to identify stroke-related symptoms and establishing a mutual referral medical support service model between lower and upper hospitals may effectively shorten the pre-hospital delay of stroke patients.

Proteogenomic Analysis and Discovery of Immune Antigens in Mycobacterium vaccae.

Tuberculosis (TB) is one of the leading causes of death worldwide, especially in developing countries. Neonatal BCG vaccination occurs in various regions, but the level of protection varies in different populations. Recently, Mycobacterium vaccae is found to be an immunomodulating therapeutic agent that could confer a significant level of protection against TB. It is the only vaccine in a phase III trial from WHO to assess its efficacy and safety in preventing TB disease in people with latent TB infection. However, the mechanism of immunotherapy of M. vaccae remains poorly understood. In this study, the full genome of M. vaccae was obtained by next-generation sequencing technology, and a proteogenomic approach was successfully applied to further perform genome annotation using high resolution and high accuracy MS data. A total of 3,387 proteins (22,508 unique peptides) were identified, and 581 proteins annotated as hypothetical proteins in the genome database were confirmed. Furthermore, 38 novel protein products not annotated at the genome level were detected and validated. Additionally, the translational start sites of 445 proteins were confirmed, and 98 proteins were validated through extension of their translational start sites based on N terminus-derived peptides. The physicochemical characteristics of the identified proteins were determined. Thirty-five immunogenic proteins of M. vaccae were identified by immunoproteomic analysis, and 20 of them were selected to be expressed and validated by Western blot for immunoreactivity to serum from patients infected with M. tuberculosis The results revealed that eight of them showed strong specific reactive signals on the immunoblots. Furthermore, cellular immune response was further examined and one protein displayed a higher cellular immune level in pulmonary TB patients. Twelve identified immunogenic proteins have orthologous in H37Rv and BCG. This is the first study to obtain the full genome and annotation of M. vaccae using a proteogenomic approach, and some immunogenic proteins that were validated by immunoproteomic analysis could contribute to the understanding of the mechanism of M. vaccae immunotherapy.

Psychological Capital and Perceived Professional Benefits: Testing the Mediating Role of Perceived Nursing Work Environment Among Chinese Nurses.

The current descriptive cross-sectional study aimed to explore the mediating role of perceived nursing work environment (PNWE) in the relationship between psychological capital (PsyCap) and perceived professional benefits among Chinese nurses. Participants (N = 351) working in two large general hospitals in Guangdong, China completed self-report questionnaires from March to May 2017. Linear regression analyses and structural equation modeling were performed to explore the mediating effect. PsyCap (particularly for hope and optimism) had a positive effect on perceived professional benefits, and PNWE was a mediator in this relationship among Chinese nurses. A good working environment can be regarded as a mediator variable, increasing staff's competence and sense of belonging to a team. For successful implementation, nurse managers should use effective strategies to increase nurses' confidence and hope while providing a comfortable work environment. [Journal of Psychosocial Nursing and Mental Health Services, 56(4), 38-47.].

[Preparation and Characterization of Pharmaceutical Properties of Tanshinone â ¡_A Microspheres].

Objective: To optimize the polymer material and process condition for preparation of tanshinone Ⅱ_A microspheres by orthogonal design. Methods: The microspheres were prepared by emulsion solvent evaporation method. The optimum polymer material and preparation process were clarified by the comprehensive weighted score which was evaluated with the drug loading, encapsulation efficiency, and yield. The quality characterization of the tanshinone Ⅱ_A microspheres were assayed by SEM, laser particle size analyzer, TGDSC,and XRD. Results: The drug loading and encapsulation rate of microspheres prepared by PLLA was significantly higher than that of other polymer material. The surface of TA-PLLA-MS was round with porous structure, average particle size was( 96. 95 ± 1. 7) µm, the drug loading was( 30. 43 ± 0. 04) %,the entrapment efficiency was( 82. 72 ± 1. 51) %,and the yield was( 94. 10 ± 1. 60) %. The drug crystal form was still in the microspheres from the results of TG-DSC and XRD. Conclusion: The PLLA tanshinone Ⅱ_A microspheres were prepared by emulsion solvent evaporation method which was simple,stable,and enough loaded drug.

Proteogenomic Analysis of Trichophyton rubrum Aided by RNA Sequencing.

Infections caused by dermatophytes, Trichophyton rubrum in particular, are among the most common diseases in humans. In this study, we present a proteogenomic analysis of T. rubrum based on whole-genome proteomics and RNA-Seq studies. We confirmed 4291 expressed proteins in T. rubrum and validated their annotated gene structures based on 35 874 supporting peptides. In addition, we identified 323 novel peptides (not present in the current annotated protein database of T. rubrum) that can be used to enhance current T. rubrum annotations. A total of 104 predicted genes supported by novel peptides were identified, and 127 gene models suggested by the novel peptides that conflicted with existing annotations were manually assigned based on transcriptomic evidence. RNA-Seq confirmed the validity of 95% of the total peptides. Our study provides evidence that confirms and improves the genome annotation of T. rubrum and represents the first survey of T. rubrum genome annotations based on experimental evidence. Additionally, our integrated proteomics and multisourced transcriptomics approach provides stronger evidence for annotation refinement than proteomic data alone, which helps to address the dilemma of one-hit wonders (uncertainties supported by only one peptide).

Novel SARS-like betacoronaviruses in bats, China, 2011.

To clarify the evolutionary relationships among betavoronaviruses that infect bats, we analyzed samples collected during 2010-2011 from 14 insectivorous bat species in China. We identified complete genomes of 2 novel betacoronaviruses in Rhinolophus pusillus and Chaerephon plicata bats, which showed close genetic relationships with severe acute respiratory syndrome coronaviruses.

Full genome of influenza A (H7N9) virus derived by direct sequencing without culture.

An epidemic caused by influenza A (H7N9) virus was recently reported in China. Deep sequencing revealed the full genome of the virus obtained directly from a patient's sputum without virus culture. The full genome showed substantial sequence heterogeneity and large differences compared with that from embryonated chicken eggs.

Co-occurrence of amikacin-resistant and -susceptible Mycobacterium tuberculosis isolates in clinical samples from Beijing, China.

OBJECTIVES: This study examined the phenomenon of heteroresistance in Mycobacterium tuberculosis clinical isolates obtained from retreated patients in Beijing, China between 2006 and 2011. METHODS: The iPLEX Gold assay platform was used to determine the prevalence of heteroresistance to injectable second-line drugs (amikacin, kanamycin and capreomycin) in resistant isolates. RESULTS: Heteroresistance was identified in 10.9% of 220 phenotypic amikacin-resistant isolates. CONCLUSIONS: Heteroresistance was related mainly to the short duration and repeated use of amikacin and capreomycin during retreatment. These findings further our understanding of the evolution of resistance to injectable drugs used for tuberculosis treatment and help guide the rational use of injectable drugs during therapy.

Combined Role of Inflammatory Biomarkers and Red Blood Cell Distribution Width in Predicting In-hospital Outcomes of Acute Ischemic Stroke Patients Undergoing Thrombolysis.

BACKGROUND: To investigate the combined effect of red blood cell distribution width (RDW) and inflammatory biomarkers on in-hospital outcomes of acute ischemic stroke(AIS) patients with thrombolysis. METHODS: 417 AIS patients with thrombolysis were included. The participants were divided into four groups according to the cut-off of white blood cell (WBC) or C reactive protein (CRP) and RDW: LWLR, LWHR, HWLR, and HWHR; or LCLR, LCHR, HCLR, and HCHR (L-low, Hhigh, W-WBC, C-CRP, R-RDW). Logistic regression models were used to calculate the odds ratios (ORs) and 95% confidence intervals (CIs) of in-hospital pneumonia and functional outcome across the four subgroups. RESULTS: Patients with higher RDW and inflammatory biomarkers levels have the highest risk of in-hospital outcomes. Compared with patients in the LWLR group, the ORs (95% CIs) of those in the HWHR group were 12.16 (4.21-35.14) and 9.31 (3.19-27.17) for in-hospital pneumonia and functional outcome. The ORs (95% CIs) of those in the HCHR group were 6.93 (2.70-17.78) and 3.38 (1.10-10.39) for in-hospital pneumonia and functional outcome, compared with patients in the LCLR group. Simultaneously adding RDW and WBC or CRP to the basic model with established risk factors significantly improved risk discrimination and reclassification for pneumonia and functional outcome (all p <0.05). CONCLUSIONS: Combined RDW and inflammatory biomarkers within 4.5 hours had a better predictive power for in-hospital outcomes of AIS patients with thrombolysis.

A comprehensive survey of bat sarbecoviruses across China in relation to the origins of SARS-CoV and SARS-CoV-2.

SARS-CoV and SARS-CoV-2 have been thought to originate from bats. In this study, we screened pharyngeal and anal swabs from 13 064 bats collected between 2016 and 2021 at 703 locations across China for sarbecoviruses, covering almost all known southern hotspots, and found 146 new bat sarbecoviruses. Phylogenetic analyses of all available sarbecoviruses show that there are three different lineages-L1 as SARS-CoV-related CoVs (SARSr-CoVs), L2 as SARS-CoV-2-related CoVs (SC2r-CoVs) and novel L-R (recombinants of L1 and L2)-present in Rhinolophus pusillus bats, in the mainland of China. Among the 146 sequences, only four are L-Rs. Importantly, none belong in the L2 lineage, indicating that circulation of SC2r-CoVs in China might be very limited. All remaining 142 sequences belong in the L1 lineage, of which YN2020B-G shares the highest overall sequence identity with SARS-CoV (95.8%). The observation suggests endemic circulations of SARSr-CoVs, but not SC2r-CoVs, in bats in China. Geographic analysis of the collection sites in this study, together with all published reports, indicates that SC2r-CoVs may be mainly present in bats of Southeast Asia, including the southern border of Yunnan province, but absent in all other regions within China. In contrast, SARSr-CoVs appear to have broader geographic distribution, with the highest genetic diversity and sequence identity to human sarbecoviruses along the southwest border of China. Our data provide the rationale for further extensive surveys in broader geographical regions within, and beyond, Southeast Asia in order to find the most recent ancestors of human sarbecoviruses.

Subpopulation analysis of heteroresistance to fluoroquinolone in Mycobacterium tuberculosis isolates from Beijing, China.

The presence of heteroresistance was represented by 23% of 235 fluoroquinolone (FQ)-resistant Mycobacterium tuberculosis isolates in Beijing, China, from 2008 to 2010. The main mechanism of FQ heteroresistance is due to the segregation of a single M. tuberculosis strain in patients; the majority of isolates with multidrug-resistant tuberculosis contained a mixture of bacterial subpopulations consisting of various mutant types, suggesting that the improper use of FQ is the major cause of FQ resistance.

Complete genome sequence of the neonatal-meningitis-associated Escherichia coli strain CE10.

Neonatal bacterial meningitis continues to be an important cause of mortality and morbidity worldwide. Escherichia coli possessing the K1 capsular polysaccharide is the most common Gram-negative pathogen causing neonatal meningitis. Here we present the complete genome sequence of neonatal meningitis-associated E. coli strain CE10, a unique K1 strain with a functional type III secretion system. Functional analysis of the genome should enhance our knowledge of the pathogenesis of neonatal E. coli K1 meningitis.

Genetic Characteristics of Human Parainfluenza Virus Types 1-4 From Patients With Clinical Respiratory Tract Infection in China.

Human parainfluenza viruses (HPIV1-4) cause acute respiratory tract infections, thereby impacting human health worldwide. However, there are no current effective antivirals or licensed vaccines for infection prevention. Moreover, sequence information for human parainfluenza viruses (HPIVs) circulating in China is inadequate. Therefore, to shed light on viral genetic diversity and evolution, we collected samples from patients infected with HPIV1-4 in China from 2012 to 2018 to sequence the viruses. We obtained 24 consensus sequences, comprising 1 for HPIV1, 2 for HPIV2, 19 for HPIV3, and 2 for HPIV4A. Phylogenetic analyses classified the 1 HPIV1 into clade 2, and the 2 HPIV4 sequences into cluster 4A. Based on the hemagglutinin-neuraminidase (HN) gene, a new sub-cluster was identified in one of the HPIV2, namely G1c, and the 19 HPIV3 sequences were classified into the genetic lineages of C3f and C3a. The results indicated that HPIV1-4 were co-circulated in China. Further, the lineages of sub-cluster C3 of HPIV3 were co-circulated in China. A recombination analysis indicated that a putative recombination event may have occurred in the HN gene of HPIV3. In the obtained sequences of HPIV3, we found that two amino acid substitution sites (R73K in the F protein of PUMCH14028/2014 and A281V in the HN protein of PUMCH13961/2014) and a negative selection site (amino acid position 398 in the F protein) corresponded to the previously reported neutralization-related sites. Moreover, amino acid substitution site (K108E) corresponded to the negative selection site (amino acid position 108) in the 10 F proteins of HPIV3. However, no amino acid substitution site corresponded to the glycosylation site in the obtained HPIV3 sequences. These results might help in studying virus evolution, developing vaccines, and monitoring HPIV-related respiratory diseases.