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Background: Current management practices and outcomes in weaning from invasive mechanical ventilation are poorly understood. We aimed to describe the epidemiology, management, timings, risk for failure, and outcomes of weaning in patients requiring at least 2 days of invasive mechanical ventilation. Methods: WEAN SAFE was an international, multicentre, prospective, observational cohort study done in 481 intensive care units in 50 countries. Eligible participants were older than 16 years, admitted to a participating intensive care unit, and receiving mechanical ventilation for 2 calendar days or longer. We defined weaning initiation as the first attempt to separate a patient from the ventilator, successful weaning as no reintubation or death within 7 days of extubation, and weaning eligibility criteria based on positive end-expiratory pressure, fractional concentration of oxygen in inspired air, and vasopressors. The primary outcome was the proportion of patients successfully weaned at 90 days. Key secondary outcomes included weaning duration, timing of weaning events, factors associated with weaning delay and weaning failure, and hospital outcomes. This study is registered with ClinicalTrials.gov, NCT03255109. Findings: Between Oct 4, 2017, and June 25, 2018, 10 232 patients were screened for eligibility, of whom 5869 were enrolled. 4523 (77·1%) patients underwent at least one separation attempt and 3817 (65·0%) patients were successfully weaned from ventilation at day 90. 237 (4·0%) patients were transferred before any separation attempt, 153 (2·6%) were transferred after at least one separation attempt and not successfully weaned, and 1662 (28·3%) died while invasively ventilated. The median time from fulfilling weaning eligibility criteria to first separation attempt was 1 day (IQR 0–4), and 1013 (22·4%) patients had a delay in initiating first separation of 5 or more days. Of the 4523 (77·1%) patients with separation attempts, 2927 (64·7%) had a short wean (≤1 day), 457 (10·1%) had intermediate weaning (2–6 days), 433 (9·6%) required prolonged weaning (≥7 days), and 706 (15·6%) had weaning failure. Higher sedation scores were independently associated with delayed initiation of weaning. Delayed initiation of weaning and higher sedation scores were independently associated with weaning failure. 1742 (31·8%) of 5479 patients died in the intensive care unit and 2095 (38·3%) of 5465 patients died in hospital. Interpretation: In critically ill patients receiving at least 2 days of invasive mechanical ventilation, only 65% were weaned at 90 days. A better understanding of factors that delay the weaning process, such as delays in weaning initiation or excessive sedation levels, might improve weaning success rates. Funding: European Society of Intensive Care Medicine, European Respiratory Society.

In this research, a NIR II luminescence imaging and enhanced chemo-/photothermal therapy system of CuS-DOX-Nd/FA NPs for breast cancer and lymph node tracing under single 808 nm irradiation is proposed. Nd-DTPA molecular cluster with the NIR II imaging effect as the carrier was designed to load the ultrasmall CuS nanoparticles and chemotherapeutic drug doxorubicin hydrochloride (DOX). The composite probe is used for tumor lesion imaging and tracking the breast cancer sentinel lymph nodes with simultaneous chemo-/photothermal therapy (PTT) for breast cancer under the single 808 nm laser. This designed probe not only has high permeability and retention (EPR) targeting effect but also can respond to the tumor microenvironment (TME), realizing more precise and efficient release of DOX at the cancer focus. At the same time, CuS as a drug carrier has a good photothermal therapy effect (photothermal conversion efficiency: 27.9%). The serialized released chemotherapy DOX and synergistic PTT effect can be used to the treat the in situ breast cancer land and simultaneously kill the metastasis cancer. The system made the combined molecular clusters Nd-DTPA achieve NIR II imaging of tumor lesions of breast cancer and lymph node to obtain the integration of diagnosis of the transferred disease for better prognosis. The feasibility of the system had obvious tumor growth inhibition effect with NIR II imaging guided is verified by a series of in vitro and in vivo experiments.

The development of eco-friendly connection material instead of steel is a challenging problem in timber structures. Following densification, the mechanical properties of low-density species can be significantly improved. Densified wood may be a potential connection material in timber structures. This paper reviewed the different processing for densified wood, and obtained favorable mechanical properties and dimensional stability based on small specimen sizes, which are much less than the applicable sizes in practice. A densification processing with alkali pretreatment was adopted for poplar widely cultivated in the world to produce the densified poplar, which has been rarely reported as connection material. Various specimens of densified poplar were tested to obtain their main mechanical properties such as strength and deformability. The set recovery of densified poplar was also measured to observe their dimensional stability. In addition, the hygroscopic swelling strains for the diameter of densified poplar dowel were measured to present their moisture-dependent behavior. The improved mechanical properties and dimensional stability confirmed the fact that densified poplar with alkali pretreatment can be an optimal connection material.

Early diagnosis is critical and challenging for tongue squamous cell carcinoma (TSCC), which is a kind of tumor with high malignancy, poor prognosis, and a high incidence of invasion and metastasis. In this research, dual-modal optical imaging rare earth nanoparticle (RENP) probes with peptide functionalization are designed for targeted TSCC imaging and therapy. RENP@C@Au (UCA) with enhanced red upconversion luminescence (UCL) and near infrared II (NIR II) imaging intensity is designed by metal and codopant modulation, and the two peaks at about 650 nm and 1064 nm of Nd ions are relatively stable with less quenching and suitable for luminescence bioimaging. Then, the cMBP peptide targeted to Cal 27 TSCC cells with highly expressed c-MET proteins is combined with UCA. Compared with human's normal cells of MCF-10A, other tumor cell lines such as A549 and HeLa, together with mice tumor cells of 4T1, the designed probe has targeted imaging and therapy to Cal 27. The final in vivo experiment shows that the probe with high NIR II luminescence signals is not easy to retain when injected into the tail vein, indicating its potential precise clinical application for the diagnosis and therapy of TSCC.

Heated humidified high-flow oxygen therapy, administered using a nasal cannula, is called high-flow nasal cannula (HFNC). HFNC has been introduced and is used as a respiratory support mode in patie...

A numerical methodology based on the coupling of computational fluid dynamics (CFD) and computational structural dynamics is established to obtain the trimming characteristics of flexible aircrafts in this paper. Reynolds-averaged Navier–Stokes equations are solved through CFD technique. Based on the frame of unstructured mesh, techniques of dynamic chimera mesh and morphing mesh are adopted to treat the data transfer between different computational zones and structure deformation caused by aeroelasticity, respectively. When it is applied to a projectile model with large slenderness ratio constructed in this paper, convergence histories of various initial conditions demonstrate the efficiency and robustness of the algorithm. The influence of the structural rigidity and normal loads on the trimming condition of flexible projectiles is investigated, and the locations of the aerodynamic center with various rigidities present the explanation that elastic deformation can move the aerodynamic center forward and weaken the margin of the stability. Furthermore, the trimming condition of flexible projectiles with propulsion is researched, which indicates that thrust misalignment will increase the effect of elastic deformation on the trimming condition, and the stability margin will be further weakened because of thrust misalignment. The conclusion provided in this paper can provide guidance for the structural design, control system design, and stability analysis for modern aircrafts with small stability margin and low rigidity.

N16 is an active protein existing in Pinctada martensi. Our previous studies have demonstrated that N16 inhibited osteoclast differentiation in vitro. To better understand how N16 regulates osteoclast differentiation, RAW264.7 cells, a murine monocytic cell line and murine bone marrow-derived macrophages (BMMs) were adopted. Treatment of RAW264.7 cells with RANKL activated osteoclastogenesis and N16 inhibited the formation of multinucleated osteoclasts and TRAP activity. The suppression occurred at the early stage of osteoclastogenesis. Moreover, we found that N16 inhibited PU.1 and MITF expressions, mirroring the inhibition of RANK expressions, indicating that N16 inhibited RANK expression by down-regulating the expressions of MITF and PU.1, thus preventing osteoclastogenesis.

In this research, a new type of mitochondrial-imaging probe using spindle nanocomposite with adjustable aspect ratios and enhanced upconversion luminescent (UCL) property was designed for synergist...

In this research, degradable peptide-modified upconversion nanoparticles (ZUPEA) were designed for the NIR II imaging and upconversion luminescence (UCL) guided photodynamic therapy (PDT) of triple-negative breast cancer (TNBC). Ultra-small rare-earth nanoparticles (RENPs) and the polymer mPEG-PLGA are polymerized into nano-microspheres via a double emulsion synthesis method, and a photosensitizer molecule (ZnPc) is added during the polymerization process to generate ZUPEA. Under 980 nm excitation, this strategy enhanced the red emission at 650 nm, showing an energy transfer efficiency of 38.3%, and the designed RENPs have better NIR II imaging abilities with a core@shell structure. These ZUPEA nanoparticles have good photodynamic therapeutic effects in vitro, and they can be degraded into small nanoparticles with a size of less than 6 nm. The cMBP-peptide-modified luminescent probe can recognize MDA-MB-231 TNBC cells in vivo when intravenously injected due to the positive targeted imaging effects of the cMBP peptide toward MET and negative targeted imaging effects relating to enhanced permeability and retention (EPR ). This specially designed ZUPEA probe with integrated diagnosis and treatment functionality provides new ideas and prospects for the use of rare-earth nanoparticles in the clinical treatment of tumors.

Objectives―To explore the potential significance of the reverberation of calcification by comparing both intravascular ultrasound (IVUS) and optical coherence tomography (OCT) measurement post manual coregistration.Background―The reverberation phenomenon is often detected by IVUS for severe calcified lesions post rotational atherectomy (RA), which is thought to be due to the glassy and smooth inner surfaces of calcifications. Because of the poor penetration of IVUS, it is impossible to measure the thickness of calcifications, and the relationship between multiple reverberations and the thickness of calcification lesions has not been reported before.Methods―A total of forty-nine patients with severe calcified coronary lesions that were detected by IVUS and OCT simultaneously were enrolled in our retrospective study. If reverberation phenomena were detected by IVUS, intravascular imaging (IVI) data (including distance between the IVUS catheter center and the inner surface of the reverberation signal, the intervals between all adjacent reverberation signals, the number of layers of reverberation in IVUS, and the thickness of the calcification in OCT) were measured at the same position and same direction (each cross-section had 4 mutually perpendicular directions) at 1-mm intervals. If the observational point (direction) had a reverberation signal, it was an effective observational point that would be enrolled in our study; otherwise, it would be regarded as an invalid observational point and excluded from this study. The correlation between each reverberation observational value and OCT data was the primary target in this retrospective study. Results―Four hundred twenty-eight valid observational points were analyzed simultaneously by IVUS and OCT; among them, 300 points had a single layer of reverberation, 83 had double layers of reverberation and 42 had multiple layers (≥3 layers) of reverberation by IVUS detection post-RA. Multivariate logistic regression analysis showed that the number of layers of reverberation by IVUS was significantly related to the thickness of calcifications by OCT at the same point and in the same direction (pConclusions―The number of layers of reverberation signal detected by IVUS is positively correlated with the thickness of calcifications measured by OCT post-RA.

Naringin and its aglycone, naringenin, occur naturally in our regular diet and traditional Chinese medicines. This study aimed to detect an effective therapeutic approach for cough variant asthma (CVA) through evaluating the relaxant effect of these two bioactive herbal monomers as antitussive and antiasthmatic on rat tracheal smooth muscle. The relaxant effect was determined by measuring muscular tension with a mechanical recording system in rat tracheal rings. Cytosolic Ca2+ concentration was measured using a confocal imaging system in primary cultured tracheal smooth muscle cells. In rat tracheal rings, addition of both naringin and naringenin could concentration dependently relax carbachol (CCh)-evoked tonic contraction. This epithelium-independent relaxation could be suppressed by BaCl2, tetraethylammonium, and iberiotoxin (IbTX), but not by glibenclamide. After stimulating primary cultured tracheal smooth muscle cells by CCh or high KCl, the intracellular Ca2+ increase could be inhibited by both naringin and naringenin, respectively. This reaction was also suppressed by IbTX. These results demonstrate that both naringin and naringenin can relax tracheal smooth muscle through opening big conductance Ca2+-activated K+ channel, which mediates plasma membrane hyperpolarization and reduces Ca2+ influx. Our data indicate a potentially effective therapeutic approach of naringin and naringenin for CVA.

The BAF complex plays an important role in the development of a wide range of tissues by modulating gene expression programs at the chromatin level. However, its role in neural crest development has remained unclear. To determine the role of the BAF complex, we deleted BAF155/BAF170, the core subunits required for the assembly, stability, and functions of the BAF complex in neural crest cells (NCCs). Neural crest-specific deletion of BAF155/BAF170 leads to embryonic lethality due to a wide range of developmental defects including craniofacial, pharyngeal arch artery, and OFT defects. RNAseq and transcription factor enrichment analysis revealed that the BAF complex modulates the expression of multiple signaling pathway genes including Hippo and Notch, essential for the migration, proliferation, and differentiation of the NCCs. Furthermore, we demonstrated that the BAF complex is essential for the Brg1-Yap-Tead-dependent transcription of target genes in NCCs. Together, our results demonstrate an important role of the BAF complex in modulating the gene regulatory network essential for neural crest development., Author summary Neural crest cells (NCCs) are a multipotent and migratory cell population that is induced at the neural plate border during neurulation and contributes to the formation of a wide range of tissues. Defects in the development, differentiation, or migration of NCCs lead to various birth defects including craniofacial and heart anomalies. Here, by genetically deleting BAF155/BAF170, the core subunits required for the assembly, stability, and functions of the BAF chromatin remodeling complex, we demonstrate that the BAF complex is essential for the proliferation, survival, and differentiation of the NCCs. Neural crest-specific deletion of BAF155/BAF170 leads to embryonic lethality due to a wide range of developmental defects including craniofacial and cardiovascular defects. By performing RNAseq and transcription factor enrichment analysis we show that the BAF complex modulates the expression of multiple signaling pathway genes including Hippo and Notch, essential for the development of the NCCs. Furthermore, the BAF complex component physically interacts with the Hippo signaling components in NCCs to regulate gene expression. We demonstrated that the BAF complex is essential for the Brg1-Yap-Tead-dependent transcription of target genes in NCCs. Together, our results demonstrate a critical role of the BAF complex in modulating the gene regulatory network essential for the proper development of neural crest and neural crest-derived tissues.

Head and neck squamous cell carcinomas (HNSCCs) are one of the most common cancers with poor survival rates, which is attributed to the difficulty in the early detection of disease. However, conventional imaging methods lack accuracy and sensitivity in the early diagnosis of HNSCCs. Thus, there is an urgent need to develop an effective and sensitive approach for HNSCC imaging. As known, the cMet receptor is overexpressed in HNSCC tumor cells CAL27 and tumor tissues. Herein, we synthesize the dual-modal near-infrared II (NIR II) imaging of luminescence and T1 magnetic resonance imaging (MRI)-based nanoprobes with the cMet targeting binding peptide (NaGdF4-PEG-cMBP), which has strong upconversion/NIR II luminescence and higher R1 relaxivity compared with the commercially used gadolinium acid (5.871 vs 3.471 mM-1 s-1). Additionally, the luminescence imaging of Yb,Er,Ce-doped probes showed that the material can efficiently accumulate in HNSCC tumors with the cMet-targeted. It can be clearly visualized in both subcutaneous and orthotopic HNSCC tumor models by dual-modal T1-weighted MRI and NIR II luminescence imaging methods. The results demonstrate that our cMBP-conjugated nanoplatform may provide a novel and very efficient noninvasive diagnostic approach for HNSCC in the near future.

In this research, the antibody of the searched hub genes has been proposed to combine with a rare-earth composite for an upconversion luminescence (UCL) and downconversion (DCL) NIR-II imaging strategy for the diagnosis of lung adenocarcinoma (LUAD). Weighted gene co-expression network analysis is used to search the most relevant hub genes, and the required top genes that contribute to tumorigenesis (negative: CLEC3B, MFAP4, PECAM1, and FHL1; positive: CCNB2, CDCA5, HMMR, and TOP2A) are identified and validated by survival analysis and transcriptional and translational results. Meanwhile, fluorescence imaging probes (NaYF4:Yb,Er,Eu@NaYF4:Nd, denoted as NYF:Eu NPs) with multimodal optical imaging properties of downconversion and upconversion luminescence in the visible region and luminescence in the near infrared II region are designed with various uniform sizes and enhanced penetration and sensitivity. Finally, when the NYF:Eu NP probe is combined with antibodies of these chosen positive hub genes (such as, TOP2A and CCNB2), the in vitro and in vivo animal experiments (flow cytometry, cell counting kit-8 assay using A549 cells, and in vivo immunohistochemistry IHC microscopy images of LUAD from patient cases) indicate that the designed nanoprobes can be excellently used as a targeted optical probe for future accurate diagnosis and surgery navigation of LUAD in contrast with other cancer cells and normal cells. This strategy of antibodies combined with optical probes provides a dual-modal luminescence imaging method for precise medicine.

Background. Pancreatic cancer is one of the most malignant tumors of the digestive system, and its treatment has rarely progressed for the last two decades. Studies on m6A regulators for the past few years have seemingly provided a novel approach for malignant tumor therapy. m6A-related factors may be potential biomarkers and therapeutic targets. This research is focused on the gene characteristics and clinical values of m6A regulators in predicting prognosis in pancreatic cancer. Methods. In our study, we obtained gene expression profiles with copy number variation (CNV) data and clinical characteristic data of 186 patients with pancreatic cancer from The Cancer Genome Atlas (TCGA) portal. Then, we determined the alteration of m6a regulators and their correlation with clinicopathological features using the log-rank tests, Cox regression model, and chi-square test. Additionally, we validated the prognostic value of m6A regulators in the International Cancer Genome Consortium (ICGC). Results. The results suggested that pancreatic cancer patients with ALKBH5 CNV were associated with worse overall survival and disease-free survival than those with diploid genes. Additionally, upregulation of the writer gene ALKBH5 had a positive correlation with the activation of AKT pathways in the TCGA database. Conclusion. Our study not only demonstrated genetic characteristic changes of m6A-related genes in pancreatic cancer and found a strong relationship between the changes of ALKBH5 and poor prognosis but also provided a novel therapeutic target for pancreatic cancer therapy.

Noise induced hearing loss (NIHL) is a kind of hearing impairment, which is next to the age-related hearing loss. More and more evidences have verified that overproduction of reactive oxygen species is a common pathologic phenomenon of different inner ear injury including NIHL, and autophagy contributes to attenuate NIHL by reducing oxidative stress. However, the underlying mechanism by which noise exposure causes autophagy activation remains unclear. In this study, we found that NIHL was accompanied by autophagy in the rat cochlea. Furthermore, twelve common genes were found at the GEO datasets GSE85290 and GSE8342, and E3 ubiquitination ligase FBXO3 was confirmed significantly reduced in NIHL rat cochlea. Next, we demonstrated that FBXO3 can directly binding with autophagy-related protein 10 (ATG10), which is necessary for the initiation of autophagy, and mediate its degradation. In vivo animal model treatment with rapamycin, an autophagy activator, significantly reduced the NIHL. Based on these data, we confirmed that FBXO3 played an important role in autophagy caused by NIHL, may be a potential target of NIHL treatment.

Background: Although Pancreatic neuroendocrine tumors(PNETs) considered as indolent tumors, most patients are diagnosed at an advanced stage. Herein, we aimed to establish a nomogram to predict the survival of PNETs patients for clinical use via Surveillance, Epidemiology, and End Results (SEER) database. Methods: Based on the SEER program, the data of 1103 patients with PNETs were enrolled and randomly divided into training set and validation set. We performed Kaplan-Meier analysis, Cox proportional hazard regression analysis in training set to evaluate the value of prognostic factors. A nomogram was constructed obtained these independent prognostic factors for predicting overall survival(OS) and specific-cancer survival(CSS). C-index, calibration curve, decision curve analysis were used to evaluate the predictive accuracy of the nomogram. Results: Age, primary site, TNM stage, grade, and surgery were associated with OS and CSS in the multivariate models. Nomograms were established depend on these risk factors and had a better discrimination power than TMN stage. The validation technologies showed that the nomogram was able to predict 3- and 5-year OS and CSS accurately, and also proved the superiority. Age, primary site, TNM stage, grade, and surgery were associated with OS and CSS in the multivariate models. Conclusions: Nomograms were established depend on these risk factors and had a better discrimination power than TMN stage. The validation technologies showed that the nomograms were able to predict 3- and 5-year OS and CSS accurately, and also proved the superiority.

Background: Pancreatic cancer is one of the most malignant tumors of the digestive system and its treatment has rarely progressed for the last two decades. Studies on m6A regulators for the past few years have seemingly provided a novel approach for malignant tumor therapy. m6A-related factors may be potential biomarkers and therapeutic targets. This research is focused on the gene characteristics and clinical values of m6A regulators in predicting prognosis in pancreatic cancer. Methods: In our study, we obtained gene expression profiles with copy number variation (CNV) data and clinical characteristic data of 186 patients with pancreatic cancer from The Cancer Genome Atlas portal (TCGA). Then, we determined the alteration of m6a regulators and their correlation with clinicopathological features using the log-rank tests, Cox regression model, and chi-square test. Results: The results suggested that pancreatic cancer patients with ALKBH5 CNV were associated with worse overall survival and disease-free survival than those with diploid genes. Additionally, upregulation of the writer gene ALKBH5 had a positive correlation with the activation of AKT pathways. Conclusion: Our study not only demonstrated genetic characteristic changes of m6A-related genes in pancreatic cancer and found a strong relationship between the changes of ALKBH5 and poor prognosis but also provided a novel therapeutic target for pancreatic cancer therapy.

Summary: Hypertrophic cardiomyopathy (HCM) is a well-established risk factor for cardiovascular mortality worldwide. Although hypertrophy is traditionally regarded as an adaptive response to physiological or pathological stress, prolonged hypertrophy can lead to heart failure. Here we demonstrate that Prdm16 is dispensable for cardiac development. However, it is required in the adult heart to preserve mitochondrial function and inhibit hypertrophy with advanced age. Cardiac-specific deletion of Prdm16 results in cardiac hypertrophy, excessive ventricular fibrosis, mitochondrial dysfunction, and impaired metabolic flexibility, leading to heart failure. We demonstrate that Prdm16 and euchromatic histone-lysine N-methyltransferase factors (Ehmts) act together to reduce expression of fetal genes reactivated in pathological hypertrophy by inhibiting the functions of the pro-hypertrophic transcription factor Myc. Although young Prdm16 knockout mice show normal cardiac function, they are predisposed to develop heart failure in response to metabolic stress. Our study demonstrates that Prdm16 protects the heart against age-dependent cardiac hypertrophy and heart failure.

In this research, a pH-sensitive degradable nanoprobe was designed by combining hydrophobic rare earth nanoparticles with biocompatible mPEG-PLGA nanomicelles for near infrared II (NIR-II) imaging-guided anti-tumor chemotherapy. The as-synthesized nanoprobes (about 300 nm) with a highly enhanced permeability and retention (EPR) effect show great potential in the diagnosis of solid tumors, providing new prospects for clinical tumor diagnosis. Then, the degradable composite probes increase the imaging sensitivity of the probe and allow for the slow release of the internal anti-tumor drugs, reducing the loss of the drug during delivery. Finally, ultra-small rare earth nanoparticles (about 6 nm) can be excreted after hydrolysis of the composite probe to reduce the enrichment of the inorganic nanoparticles in vivo. Thus, this degradable NIR-II imaging probe based on a polymer-lanthanide composite could be a promising candidate for preclinical cancer chemotherapy and surgery navigation under a single 808 nm laser.

Tissue-resident macrophages (TRMØs) can act as innate-immune sentinels to protect body against microbe invaders and stimulating materials such as cholesterol crystals in cholesteatoma, as well as to preserve tissue integrity by cleaning unwanted cellular debris.TRMØs in the incised middle ear tissues were obtained from the patients with cholesteatoma as an experimental group and the patients without cholesteatoma as a control group. Differential gene expression profiling of TRMØs was conducted between two groups by analyzing GO processes, KEGG and GSEA pathways of inflammation, tissue repair and homeostasis.The current study showed that 145 of 7060 genes were significantly up-regulated (logFC2 and FDR0.05) when compared with the patients without cholesteatoma. GO process, GSEA and Cytoscape analysis of the over-expressed genes illustrated the boosted inflammatory and anti-infection functions of TRMØs existed neutrophil function, leukocyte migration, and adaptive immune response involved receptors and signaling pathways. Whereas the homeostasis and repair functions of TRMØs were affected from up-regulated genes, such as over-expressed keratin-13 that helped form the outer keratinising squamous epithelial layer, and over-expressed MMPs that activated the extracellular matrix molecules to promote inflammation and disturb tissue remodeling. Additionally, 74 down-regulated genes (logFC-2 and FDR0.05) also affected the homeostasis and repair functions by affecting extracelluar matrix structure and contractile fibres in TRMØs.The cellular and molecular levels in cholesteatoma is attributable to chronic infection and several disturbed cellular biological processes involving cell integrity and tissue remodeling.

Background Hereditary spherocytosis (HS) is a type of hemolytic anemia caused by abnormal red cell membrane skeletal proteins with few unique clinical manifestations in the neonate and infant. An ANK1 gene mutation is the most common cause of HS. Case presentation The patient was a 11-month-old boy who suffered from anemia and needed a regular transfusion therapy at an interval of 2–3 months. Hematological investigations showed moderate anemia (Hb80 g/L). Red cells displayed microcytosis (MCV76.4 fl, MCH25.6 pg, MCHC335 g/L). The reticulocytes were elevated (4.8%) and the spherocytes were increased (10%). Direct antiglobulin test was negative. Biochemical test indicated a slight elevation of bilirubin, mainly indirect reacting (TBIL32.5 μmol/L, IBIL24 μmol/L). The neonatal HS ratio is 4.38, obviously up the threshold. Meanwhile, a de novo ANK1 mutation (exon 25:c.2693dupC:p.A899Sfs*11) was identified by next-generation sequencing (NGS). Thus, hereditary spherocytosis was finally diagnosed. Conclusions Gene detection should be considered in some hemolytic anemia which is difficult to diagnose by routine means. We identified a novel de novo ANK1 heterozygous frameshift mutation in a Yi nationality patient while neither of his parents carried this mutation. Electronic supplementary material The online version of this article (10.1186/s12887-019-1436-4) contains supplementary material, which is available to authorized users.

Background: Thalassemia is an autosomal genetic disorder, found throughout the world. It is still not treatable and create socio economic problems. In this study, we investigated the prevalence and spectrum features of thalassemia in Yunnan Province, the southwestern area of China. During 2014–2018, a total of 3,539 suspected thalassemia children were detected with α- and β-thalassemia mutations by gap-Polymerase Chain Reaction (PCR) and reverse dot blot (RDB) analysis in Kunming Children's Hospital.Results: Of these patients, 1,130 were diagnosed thalassemia gene carriers with a carrying rate of 31.92%. Among them, α-thalassemia was 43.63%, β-thalassemia was 53.98%, cases with both α- and β- thalassemia was 2.39%. In α-thalassemia patients, the most common mutations was –SEA/αα (52.13%), followed by –α3.7/αα (27.79%), hemoglobin H disease (18.46%), and –α4.2/αα (1.62%). Fifteen gene mutations and 30 genotypes were identified in β-thalassemia patients, with the five most common mutations CD17 (A>T) (29.51%), CD41–42 (–TTCT) (27.87%), IVS-II-654 (C>T) (14.92%), CD26 (G>A) (6.89%), and CD26/CD27 (2.62%) accounting for 81.81% of the β-globin gene mutations. Furthermore, we founded two rare mutations CD34 (TGG → TAG) and Int in Chinese populations.Conclusions: Our results suggested that the prevalence and gene mutation spectrum of thalassemia display obviously heterogeneity among children in Yunnan Province. The findings provide the valuable information for premarital and pre-pregnancy screening, prenatal diagnostic services, and designing appropriate prevention programs to control thalassemia for future in this area.

Objective. This study was aimed to investigate the protective effect of methylene blue against lung injury induced by reperfusion of ischemic hindlimb in a rat model. Methods. Twenty-four healthy adult male Sprague-Dawley rats were equally randomized into three groups: sham (SM) group, ischemia reperfusion (IR) group, and methylene blue (MB) group. Rats in both IR and MB groups were subjected to 4 h of ischemia by clamping the left femoral artery and then followed by 4 h of reperfusion. Treatment with 1% methylene blue (50 mg/kg) was administrated intraperitoneally at 10 min prior to reperfusion in the MB group. After 4 h of reperfusion, malondialdehyde (MDA) level, myeloperoxidase (MPO), and superoxide dismutase (SOD) activities in lung tissue were detected; inflammatory cytokines, including IL-1β and IL-6, were measured in bronchoalveolar lavage fluid (BALF); correspondingly, the morphological changes and water content in both gastrocnemius muscle and lung samples were evaluated. Results. Hindlimb IR caused remarkable morphological abnormalities and edema in both muscle and lung tissues. SOD activity was decreased, both the MPO activity and MDA level in lung tissue, as well as IL-1β and IL-6 levels in BALF, were increased in the IR group (p<0.05). Compared with the IR group, SOD activity was increased, whereas MPO activity and MDA level in lung tissue and IL-1β and IL-6 levels in BALF were decreased in the MB group (p<0.05). Also, the histological damage and edema in both lung and muscle tissues were significantly attenuated by the treatment of methylene blue. Conclusion. Methylene blue attenuates lung injury induced by hindlimb IR in rats, at least in part, by inhibiting oxidative stress.

In this research, an optimized phosphor combined with naturally green chlorophyll (Phosphor-Chlorophyll, denoted as Ph-chl) was designed for dual-modal luminescence-guided anti-tumor surgery and photodynamic therapy (PDT). A genetic algorithm (GA) is used to solve the “low up-conversion luminescence (UCL) and short-wavelength infrared (SWIR) intensity” problem by coding the proportions of elements in the phosphor in order to find the optimal phosphor with enhanced red UCL emission and SWIR luminescence using Yb/Er in the core and Yb/Nd in the shell. Moreover, when phosphors with different emission light colors (blue and green) are combined with chlorophyll as the control, the results indicate that phosphors with red emission as the energy donor have high PDT efficiency to activate the chlorophyll for reactive oxygen species (ROS) production. Additionally, due to the enhanced penetration and retention (EPR) effect, the as-synthesized Ph-chl could be used for surgery navigation with a higher SWIR imaging effect focusing on cancer rather than normal tissues and paracancerous tissue. Thus, the high dual-modal luminescence guided surgery properties of the final optimized phosphor will promote its further use in minimally-invasive endoscopic clinical surgery navigation.

In recent years, wearable monitoring devices have been very popular in the health care field and are being used to avoid sport injuries during exercise. They are usually worn on the wrist, the same as sport watches, or on the chest, like an electrocardiogram patch. Common functions of these wearable devices are that they use real time to display the state of health of the body, and they are all small sized. The electromyogram (EMG) signal is usually used to show muscle activity. Thus, the EMG signal could be used to determine the muscle-fatigue conditions. In this study, the goal is to develop an EMG patch which could be worn on the lower leg, the gastrocnemius muscle, to detect real-time muscle fatigue while exercising. A micro controller unit (MCU) in the EMG patch is part of an ARM Cortex-M4 processor, which is used to measure the median frequency (MF) of an EMG signal in real time. When the muscle starts showing tiredness, the median frequency will shift to a low frequency. In order to delete the noise of the isotonic EMG signal, the EMG patch has to run the empirical mode decomposition algorithm. A two-electrode circuit was designed to measure the EMG signal. The maximum power consumption of the EMG patch was about 39.5 mAh. In order to verify that the real-time MF values measured by the EMG patch were close to the off-line MF values measured by the computer system, we used the root-mean-square value to estimate the difference in the real-time MF values and the off-line MF values. There were 20 participants that rode an exercise bicycle at different speeds. Their EMG signals were recorded with an EMG patch and a physiological measurement system at the same time. Every participant rode the exercise bicycle twice. The averaged root-mean-square values were 2.86 &plusmn, 0.86 Hz and 2.56 &plusmn, 0.47 Hz for the first and second time, respectively. Moreover, we also developed an application program implemented on a smart phone to display the participants&rsquo, muscle-fatigue conditions and information while exercising. Therefore, the EMG patch designed in this study could monitor the muscle-fatigue conditions to avoid sport injuries while exercising.

Acute lung injury (ALI) is characterized by tissue damage and inflammatory cytokine secretion; however, the therapeutic options available to treat ALI remain limited. Necrostatin‑1 (Nec‑1) has the ability to attenuate cell necroptosis in various inflammatory diseases. The present study evaluated the protective effects of Nec‑1 on a mouse model of lipopolysaccharide‑induced ALI. Histological alterations in the lungs were evaluated through hematoxylin and eosin staining, and the expression levels of cytokines in the bronchoalveolar lavage fluid and lung tissues were determined by ELISA. In addition, accumulated production of reactive oxygen species was determined by staining with DCFH‑DA probes, western blotting and immunofluorescence. The results revealed that treatment with the necroptosis inhibitor, Nec‑1, exerted significant protective effects on ALI‑induced inflammation and necroptosis. The key proteins involved in necroptosis were markedly reduced, including receptor‑interacting serine/threonine‑protein kinase (RIP)1 and RIP3. Notably, antioxidant proteins were upregulated by Nec‑1, which may attenuate oxidative stress. Furthermore, treatment with Nec‑1 markedly suppressed necroptosis in the pulmonary alveoli RLE‑6TN cell line. Taken together, these data revealed a novel association between ALI and necroptosis, and suggested that necroptosis inhibitors may be used as effective anti‑inflammatory drugs to treat ALI.

Free fatty acid receptor G protein-coupled receptor 120 (GPR120) is highly expressed in macrophages and was reported to inhibit lipopolysaccharide (LPS)-stimulated cytokine expression. Under inflammation, macrophages exhibit striking functional changes, but changes in GPR120 expression and signaling are not known. In this study, the effects of LPS treatment on macrophage GPR120 expression and activation were investigated. The results showed that LPS inhibited GPR120 expression in mouse macrophage cell line Ana-1 cells. Moreover, LPS treatment inhibited GPR120 expression in mouse alveolar macrophages both in vitro and in vivo. The inhibitory effect of LPS on GPR120 expression was blocked by Toll-like receptor 4 (TLR4) inhibitor TAK242 and p38 mitogen-activated protein kinase inhibitor LY222820, but not by ERK1/2 inhibitor U0126 and c-Jun N-terminal kinase inhibitor SP600125. LPS-induced inhibition of GPR120 expression was not attenuated by GPR120 agonists TUG891 and GW9508. TUG891 inhibited the phagocytosis of alveolar macrophages, and LPS treatment counteracted the effects of TUG891 on phagocytosis. These results indicate that pretreatment with LPS inhibits GPR120 expression and activation in macrophages. It is suggested that LPS-induced inhibition of GPR120 expression is a reaction enhancing the LPS-induced pro-inflammatory response of macrophages.

N16 is an active protein isolated and screened from nacre proteins. Our previous studies have shown that N16 exerted a good effect on osteoporosis caused by dexamethasone on female rats. In order to further confirm the action of N16 against glucocorticoid-induced osteoporosis (GIOP) and clarify its possible mechanisms, prednisolone-treated larval zebrafish (Danio rerio) model was adopted. Administration of N16 resulted in a significant increase of vertebral bone density of osteoporotic zebrafish, indicating a good effect of N16 against GIOP. Transcriptomic sequencing analysis was then performed to investigate the mechanisms of the action of N16 against GIOP. It was observed that N16 modulated the osteoporotic phenotype by up-regulating the osteoblastic characteristic genes and down-regulating the osteoclastic characteristic genes in zebrafish.

The incorporation of the structural elements of thermostable enzymes into their less stable counterparts is generally used to improve enzyme thermostability. However, the process of engineering enzymes with both high thermostability and high activity remains an important challenge. Here, we report that the thermostability and activity of a thermophilic subtilase were simultaneously improved by incorporating structural elements of a psychrophilic subtilase. There were 64 variable regions/residues (VRs) in the alignment of the thermophilic WF146 protease, mesophilic sphericase, and psychrophilic S41. The WF146 protease was subjected to systematic mutagenesis, in which each of its VRs was replaced with those from S41 and sphericase. After successive rounds of combination and screening, we constructed the variant PBL5X with eight amino acid residues from S41. The half-life of PBL5X at 85°C (57.1 min) was approximately 9-fold longer than that of the wild-type (WT) WF146 protease (6.3 min). The substitutions also led to an increase in the apparent thermal denaturation midpoint temperature ( T m ) of the enzyme by 5.5°C, as determined by differential scanning calorimetry. Compared to the WT, PBL5X exhibited high caseinolytic activity (25 to 95°C) and high values of K m and k cat (25 to 80°C). Our study may provide a rational basis for developing highly stable and active enzymes, which are highly desired in industrial applications.

N16, a nacreous protein isolated from Pinctada martensii, is related to nacreous layer formation. Our previous study indicated that N16 showed dual regulatory effects by inducing osteoblast biomineralization as well as inhibiting osteoclast formation. In order to obtain large quantity of N16 for animal experiment and clinical trial, a fermentation and preparative purification method was established. The N16 cDNA was cloned to a BL21(DE3)plysE-pET32a vector and grown in a 20 L fermenter. The medium, temperature, pH and dissolved oxygen (DO) were optimized. N16 was expressed in inclusion bodies. It was denatured and refolded in 8 M urea buffer and purified to 97% purity by passing through a gel filtration column. The glucocorticoid induced osteoporosis (GIO) rat model was used to investigate the anti-osteoporosis activity of N16 in vivo. Results showed that the decrease of the bone mineral density (BMD) and the ultimate load was significantly relieved after N16 treatment. N16 displayed dual regulatory effects by promoting osteogenesis as well as inhibiting bone resorption in vivo. Our work will contribute to further clinical studies on N16 for osteoporosis treatment.

The proform of the WF146 protease, an extracellular subtilase produced by thermophilic Bacillus sp. WF146, matures efficiently at high temperatures. Here we report that the proform, which contains an N-terminal propeptide composed of a core domain (N*) and a linker peptide, is intrinsically able to mature via multiple pathways. One autocatalytic pathway is initiated by cis-processing of N* to generate an autoprocessed complex N*-I(WT), and this step is followed by truncation of the linker peptide and degradation of N*. Another autocatalytic pathway is initiated by trans-processing of the linker peptide followed by degradation of N*. Unlike most reported subtilases, the maturation of the WF146 protease occurs not only autocatalytically but also hetero-catalytically whereby heterogeneous proteases accelerate the maturation of the WF146 protease via trans-processing of the proform and N*-I(WT). Although N* acts as an intramolecular chaperone and an inhibitor of the mature enzyme, the linker peptide is susceptible to proteolysis, allowing the trans-processing reaction to occur auto- and hetero-catalytically. These studies also demonstrate that the WF146 protease undergoes subtle structural adjustments during the maturation process and that the binding of Ca(2+) is required for routing the proform to mature properly at high temperatures. Interestingly, under Ca(2+)-free conditions, the proform is cis-processed into a unique propeptide-intermediate complex (N*-I(E)) capable of re-synthesis of the proform. Based on the basic catalytic principle of serine proteases and these experimental results, a mechanism for the cis-processing/re-synthesis equilibrium of the proform and the role of the linker peptide in regulation of this equilibrium has been proposed.

In recent years, the use of liquid biofuels in the transport sector has shown rapid global growth, driven mostly by policies focused on achievement of energy security, and mitigation of GHG emissions. Microalgae, as biomass, are a potential source of renewable energy, and they can be converted into energy such as biofuel oil and gas.In this paper, the microalgae strain of Chlorella sp. was supplied in BG11 medium, and Cultivation optimization were experimented. The results were showed that temperature was an important factor which could influence growth and accumulation of secondary metabolite in the cell from accumulation process of biological products, temperature which was suitable for the growth of chlorella was 25°C.

OBJECTIVE To observe the effects of Shengqing Capsule (SQC), a compound of traditional Chinese herbal medicine, on biochemical parameters in C57BL/6J mice with cholesterol gallstone. METHODS Thirty-eight C57BL/6J mice were randomly divided into normal control group (n=10), untreated group (n=15) and SQC group (n=13). Cholesterol gallstone was induced in mice of the latter two groups by feeding high cholesterol diet. Mice in the SQC group were intragastricly administered SQC 0.22 g/(kg.d). After 8-week treatment, animals were sacrificed and sampled to calculate the incidences of stone formation. The contents of serum cholesterols and Ca(2+), and the malonaldehyde (MDA) content and superoxide dismutase (SOD) activity in liver tissues were detected. RESULTS The incidences of stone formation were 73.33% in untreated group, 0% in normal control group, and 23.08% in the SQC group. And the INCIDENCE in untreated group was significantly higher than those in the normal control group and the SQC group (P

Bacillopeptidase F (Bpr) is a fibrinolytic serine protease produced by Bacillus subtilis . Its precursor is composed of a signal peptide, an N-terminal propeptide, a catalytic domain, and a long C-terminal extension (CTE). Several active forms of Bpr have been previously reported, but little is known about the maturation of this enzyme. Here, a gene encoding a Bpr (BprL) was cloned from B. subtilis LZW and expressed in B. subtilis WB700, and three fibrinolytic mature forms with apparent molecular masses of 45, 75, and 85 kDa were identified in the culture supernatant. After treatment with urea, the 75-kDa mature form had the same molecular mass as the 85-kDa mature form, from which we infer that they adopt different conformations. Mutational analysis revealed that while the 85-kDa mature form is generated via heterocatalytic processing of a BprL proform by an unidentified protease of B. subtilis , the production of the 75- and 45-kDa mature forms involves both hetero- and autocatalytic events. From in vitro analysis of BprL and its sequential C-terminal truncation variants, it appears that partial removal of the CTE is required for the initiation of autoprocessing of the N-terminal propeptide, which is composed of a core domain (N*) and a 15-residue linker peptide, thereby yielding the 45-kDa mature form. These data suggest that the differential processing of BprL, either heterocatalytically or autocatalytically, leads to the formation of multiple mature forms with different molecular masses or conformations.

In the present study, we successfully isolated PDMSCs from human placental tissues. The RT-PCR results show that PDMSCs preserved the genetic characteristics of the primitive embryonic stage--Oct-4 and Nanog. By using serum-free medium supplemented essential growth factors and induction medium culture for 4 weeks, a monolayer of spindle-like PDMSCs gradually formed 3D spheroid bodies (SB-PDMSCs). By using real-time RT-PCR, early mRNA expressions of Pdx1, as well as the Sox17 and Foxa2 genes, were observed to be significantly activated in SB-PDMSCs, followed by the expression of mature pancreas-related genes (insulin, glucagon, and somatostatin). The high insulin content of SB-PDMSCs was further confirmed by ELISA assay, and the glucose dependency was demonstrated by the corresponding insulin secretion level. In a transplantation study of streptozotocin-pretreated nude mice, the restoration of normoglycemia in the SB-PDMSC treated group was further observed. In conclusion, these results indicate that PDMSCs are an excellent source for the induced differentiation of well-functioning insulin-positive cells. The potential of these insulin producing cells derived from PDMSCs was also demonstrated functionally by the demonstration of secreted insulin in vitro and effective control of blood glucose levels in vivo.

Beak is one of the most important hard tissues of Cephalopoda which is usually used in fishery biology and biomass estimation.According to the 2 916 samples of short-fin squid Illex argentinus collected by Chinese squid jigging vessels from January to March in 2010 in Southwest Atlantic Ocean,the morphologic parameters were measured and the growth characteristics of beak were studied.The result of principal component analysis of twelve morphologic indices showed that upper hood length(UHL),upper crest length(UCL),upper wing length(UWL),lower hood length(LHL),lower crest length(LCL)and lower wing length(LWL)could be used to represent the length growth features of beak of each dome,and the ratios of HL to CL,RL to RL,RW to WL,LWL/CL and WL/CL could be used as indicators of entire growth for beak(upper beak and lower beak)growth,and the each dome growth,the entire beak growth impacted by sex,gonad maturity and individual size were analyzed by the analysis of variance(ANOVA)and least-significant difference(LSD).The results indicated that the UHL,UCL,UWL,LHL,LCL and LWL were closely related to sex,and for females or males,the UHL,UCL,UWL,LHL,LCL and LWL were closely related to gonad maturity and mantle length,however the ratios of HL/CL,RL/CL,RW/CL,LWL/CL,WL/CL were nearly constant without changing with sex,gonad maturity or different mantle length with the ratios of 78.04%,22.46%,17.28%,81.41% and 23.07% for upper beak and 50.69%,43.31%,41.13%,144.98% and 93.68% for lower beak respectively.This study suggested that there are significant effects of individual size on beak morphology of I.argentinus in the Southwestern Atlantic Ocean.