A detailed analysis of 16S rRNA gene sequencing and conventional PCR-based testing for the diagnosis of bacterial pathogens and discovery of novel bacteria.

This study represents the first analysis of the bacterial community in chickens affected by swollen head syndrome, utilizing 16S rRNA gene sequencing. Samples were obtained from clinical laying chickens and were examined for the presence of Avibacterium paragallinarum (APG) and Ornithobacterium rhinotracheale (ORT) using conventional polymerase chain reaction (PCR). From the samples, five APG-positive (APG) and APG-negative (N-APG) samples were chosen, along with five specific pathogen-free chickens, for 16S rRNA gene sequencing. Results showed that APG and ORT were widely detected in the chicken samples with swollen head syndrome (SHS, 9/10), while APG was detected in all five specific pathogen-free (SPF) samples. In contrast, conventional PCR sensitivity was found to be inadequate for diagnosis, with only 35.7% (5/14) and 11.1% (1/9) sensitivity for APG and ORT, respectively, based on 16S rRNA gene sequencing data. Furthermore, 16S rRNA gene sequencing was able to quantify the bacteria in the samples, revealing that the relative abundance of APG in the APG group ranged from 2.7 to 81.3%, while the relative abundance of APG in the N-APG group ranged from 0.1 to 21.0%. Notably, a low level of APG was also detected in all 5 SPF samples. The study also identified a significant number of animal and human common bacterial pathogens, including but not limited to Gallibacterium anatis, Riemerella columbina, Enterococcus cecorum, Mycoplasma synoviae, Helicobacter hepaticus, and Staphylococcus lentus. In conclusion, 16S rRNA gene sequencing is a valuable tool for bacterial pathogen diagnosis and the discovery of novel bacterial pathogens, while conventional PCR is not reliable for diagnosis.

Detection of Dopamine Based on Aptamer-Modified Graphene Microelectrode.

In this paper, a novel aptamer-modified nitrogen-doped graphene microelectrode (Apt-Au-N-RGOF) was fabricated and used to specifically identify and detect dopamine (DA). During the synthetic process, gold nanoparticles were loaded onto the active sites of nitrogen-doped graphene fibers. Then, aptamers were modified on the microelectrode depending on Au-S bonds to prepare Apt-Au-N-RGOF. The prepared microelectrode can specifically identify DA, avoiding interference with other molecules and improving its selectivity. Compared with the N-RGOF microelectrode, the Apt-Au-N-RGOF microelectrode exhibited higher sensitivity, a lower detection limit (0.5 µM), and a wider linear range (1~100 µM) and could be applied in electrochemical analysis fields.

Integrated metabolome and transcriptome analyses of anthocyanin biosynthesis reveal key candidate genes involved in colour variation of Scutellaria baicalensis flowers.

BACKGROUND: Bright flower colour assists plants attract insects to complete pollination and provides distinct ornamental values. In some medicinal plants, diverse flower colour variations usually imply differences in active ingredients. Compared to the common bluish purple of Scutellaria baicalensis flower (SB), the natural variants present rose red (SR) and white (SW) flowers were screened out under the same growing conditions in the genuine producing area Shandong Province, China. However, the mechanism of flower colour variation in S. baicalensis was remain unclear. In the present study, we conducted integrated transcriptome and metabolome analyses to uncover the metabolic difference and regulation mechanism in three S. baicalensis flowers. RESULTS: The results showed that 9 anthocyanins were identified. Among which, 4 delphinidin-based anthocyanins were only detected in SB, 4 cyanidin-based anthocyanins (without cyanidin-3-O-glucoside) mainly accumulated in SR, and no anthocyanin but high level of flavanone, naringenin, was detected in SW. The gene expression profile indicated that the key structural genes in the flavonoid and anthocyanin biosynthesis pathway differentially expressed in flowers with different colours. Compared to SB, the down-regulated expression of F3'5'H, ANS, and 3GT gene in SR might influence the anthocyanin composition. Especially the InDel site with deletion of 7 nucleotides (AATAGAG) in F3'5'H in SR might be the determinant for lack of delphinidin-based anthocyanins in rose red flowers. In SW, the lower expression levels of DFR and two F3H genes might reduce the anthocyanin accumulation. Notably the SNP site of G > A mutation in the splicing site of DFR in SW might block anthocyanin biosynthesis from flavanones and thus cause white flowers. In addition, several key transcription factors, including MYB, bHLH, and NAC, which highly correlated with structural gene expression and anthocyanin contents were also identified. CONCLUSIONS: These results provide clues to uncover the molecular regulatory mechanism of flower colour variation in S. baicalensis and promote novel insights into understanding the anthocyanin biosynthesis and regulation.

Boron nitride nanotube-salt-water hybrid:crystalline precipitation.

Molecular dynamics simulation is used to study the transport characteristics of NaCl solution in boron nitride nanotubes (BNNTs). It presents an interesting and well-supported MD study of the crystallization of NaCl from its water solution under the confinement of a 3 nm thick boron nitride nanotube with varied surface charging conditions. The results of the molecular dynamics simulation indicate that NaCl crystallization occurs in charged BNNTs at room temperature when the concentration of NaCl solution reaches about 1.2 M. The reason for this phenomenon is as follows: when the number of ions in the nanotubes is high, the double electric layer that forms at the nanoscale near the charged wall surface, the hydrophobicity of BNNTs, and the interaction among ions cause ions to aggregate in the nanotubes. As the concentration of NaCl solution increases, the concentration of ions when they aggregate in the nanotubes reaches the saturation concentration of the NaCl solution, resulting in the crystalline precipitation phenomenon.

A prolific and robust whole-genome genotyping method using PCR amplification via primer-template mismatched annealing.

Whole-genome genotyping methods are important for breeding. However, it has been challenging to develop a robust method for simultaneous foreground and background genotyping that can easily be adapted to different genes and species. In our study, we accidently discovered that in adapter ligation-mediated PCR, the amplification by primer-template mismatched annealing (PTMA) along the genome could generate thousands of stable PCR products. Based on this observation, we consequently developed a novel method for simultaneous foreground and background integrated genotyping by sequencing (FBI-seq) using one specific primer, in which foreground genotyping is performed by primer-template perfect annealing (PTPA), while background genotyping employs PTMA. Unlike DNA arrays, multiple PCR, or genome target enrichments, FBI-seq requires little preliminary work for primer design and synthesis, and it is easily adaptable to different foreground genes and species. FBI-seq therefore provides a prolific, robust, and accurate method for simultaneous foreground and background genotyping to facilitate breeding in the post-genomics era.

Poor response to selpercatinib plus crizotinib in a rearranged during transfection fusion-positive patient with acquired selpercatinib-resistant MNNG HOS transforming amplification: a case report.

Selpercatinib has been approved by most major regulatory bodies in 2020 and become the standard therapy for rearranged during transfection ( RET )-rearranged nonsmall-cell lung cancer (NSCLC). Knowledge is limited regarding mechanisms of resistance to selpercatinib and effective treatment. One study identified MNNG HOS transforming ( MET ) amplification as intrinsic or secondary resistance mechanism from four patients, and three of them showed ~40% tumor reduction when treated with selpercatinib plus crizotinib. We report a 30-year-old female nonsmoker diagnosed in 2019 with stage IV lung adenocarcinoma harboring KIF5B-RET and a novel FOXD1-RET fusion. Frontline therapy consisted of bevacizumab combined with pemetrexed and carboplatin and achieved a progression-free survival (PFS) of 14 months with best response of stable disease. The patient then enrolled in the LIBRETTO-321 trial (NCT03157129) and started selpercatinib, which elicited a PFS of 9 months with best response of partial response. MNNG HOS transforming ( MET ) amplification was subsequently detected upon progression on selpercatinib, and the patient was placed on third-line treatment with selpercatinib plus crizotinib. However, her health deteriorated rapidly and died of cancer 4 months later. We provided additional evidence supporting MET amplification as an acquired mechanism of resistance to selective RET inhibition. In addition, the apparent lack of response to selpercatinib plus crizotinib in this case highlights the need for future cohort studies for examining the value of combining RET and MET inhibitors in treating RET -rearranged, MET -amplified NSCLC.

MiR-338-3p Is a Biomarker in Neonatal Acute Respiratory Distress Syndrome (ARDS) and Has Roles in the Inflammatory Response of ARDS Cell Models.

To investigate the association between serum miR-338-3p levels and neonatal acute respiratory distress syndrome (ARDS) and its mechanism. The relative miR-338-3p expression in serum was detected by quantitative real-time RT-PCR. Interleukin-1beta (IL-1ß), IL-6, and tumor necrosis factor-alpha (TNF-α) levels were detected by ELISAs. A receiver operating characteristic (ROC) curve analysis of serum miR-338-3p evaluated the diagnosis of miR-338-3p in neonatal ARDS. Pearson's correlation analysis evaluated the correlation between serum miR-338-3p and neonatal ARDS clinical factors. Flow cytometry evaluated apoptosis, and a CCK-8 assay assessed cell viability. A luciferase assay evaluated the miR-338-3p/AKT3 relationship. The miR- 338-3p expression was decreased in neonatal ARDS patients and in lipopolysaccharide (LPS)-treated cells. The ROC curve showed the accuracy of miR-338-3p for evaluating neonatal ARDS patients. The correlation analysis demonstrated that miR-338-3p was related to PRISM-III, PaO2/FiO2, oxygenation index, IL-1ß, IL-6, and TNF-α in neonatal ARDS patients. MiR-338-3p overexpression inhibited the secretion of inflammatory components, stifled cell apoptosis, and LPS-induced advanced cell viability. The double-luciferase reporter gene experiment confirmed that miR-338-3p negatively regulates AKT3 mRNA expression. Serum miR-338-3p levels were related to the diagnosis and severity of neonatal ARDS, which may be attributed to its regulatory effect on inflammatory response in ARDS.

Recognition of Underwater Materials of Bionic and Natural Fishes Based on Blue-Green Light Reflection.

Thanks to the advantages of low disturbance, good concealment and high mobility, bionic fishes have been developed by many countries as equipment for underwater observation and data collection. However, differentiating between true and bionic fishes has become a challenging task. Commonly used acoustic and optical technologies have difficulty in differentiating bionic fishes from real ones due to their high similarity in shape, size, and camouflage ability. To solve this problem, this paper proposes a novel idea for bionic fish recognition based on blue-green light reflection, which is a powerful observation technique for underwater object detection. Blue-green light has good penetration under water and thus can be used as a signal carrier to recognize bionic fishes of different surface materials. Three types of surface materials representing bionic fishes, namely titanium alloy, carbon fiber, and nylon, are investigated in this paper. We collected 1620 groups of blue-green light reflection data of these three kinds of materials and for two real fishes. Following this, three machine learning algorithms were utilized for recognition among them. The recognition accuracy can reach up to about 92.22%, which demonstrates the satisfactory performance of our method. To the best of our knowledge, this is the first work to investigate bionic fish recognition from the perspective of surface material difference using blue-green light reflection.

Relationships between self-efficacy, coping style and quality of work-life among nursing managers in China: A cross-sectional study.

AIM: To investigate the quality of work-life among nursing managers in China and analyse the relationships between self-efficacy, coping style and quality of work-life. BACKGROUND: Self-efficacy, coping-style and quality of work-life play significant roles in the stability of nursing teams, the quality of clinical care and patient safety. Although some factors influencing quality of work-life have been identified, there is no large-scale study of the relationships among self-efficacy, coping style and quality of work-life of nursing managers. Strategies to strengthen the quality of work-life of nursing managers should be developed by exploring the relationships between the three variables. METHODS: A nationwide cross-sectional study was conducted. A total of 1498 nursing managers from 51 tertiary general hospitals in 20 provinces in China were selected for the study using multistage stratified proportional sampling. Registered full-time nursing managers with more than 1 year of management experience were eligible for the study. The participants completed online questionnaires to assess their self-efficacy, coping style and work-related quality of life. The data were analysed using descriptive analysis, Pearson's correlation, hierarchical multiple linear regression and structural equation modelling to verify the relationships between variables. RESULTS: The quality of work-life score for the nursing managers surveyed was 3.74 ± 0.56 out of a possible 5. Self-efficacy was positively correlated with quality of work-life (ß = .484, P < .01) over all demographic variables. Positive coping was also positively associated with quality of work-life (ß = .404, P < .01). Self-efficacy and positive coping explained 22.7% and 14.2% of the variance in the quality of work-life, respectively. Structural equation modelling indicated that self-efficacy positively and directly predicted the quality of work-life (ß = .395, P < .001), and indirectly affected quality of work-life via positive coping (ß = .186, P < .001), but not via negative coping (ß = .005, P > .05). The final model could explain 44% of the variance in the quality of work-life. CONCLUSION: Self-efficacy was positively correlated with quality of work-life among Chinese nursing managers, and positive coping partially mediated this relationship. The results suggest that self-efficacy and positive coping are pivotal facilitators for improving the quality of work-life and need to be taken into account in measures to improve the quality of work-life of nursing managers. IMPLICATIONS FOR NURSING MANAGEMENT: Hospital administrators should recognize the importance of nursing managers' work-life quality to promote both the stability of nursing teams and the quality of clinical care and safety provided. They should help nursing managers foster self-efficacy and active coping abilities through incentive measures and provide positive coping training courses to enhance their quality of work-life.

OsPPR939, a nad5 splicing factor, is essential for plant growth and pollen development in rice.

KEY MESSAGE: P-subfamily PPR protein OsPPR939, which can be phosphorylated by OsS6K1, regulates plant growth and pollen development by involving in the splicing of mitochondrial nad5 introns 1, 2, and 3. In land plants, pentatricopeptide repeat (PPR) proteins play key roles in mitochondrial group II intron splicing, but how these nucleus-encoded proteins are imported into mitochondria is unknown. To date, a few PPR proteins have been characterized in rice (Oryza sativa). Here, we demonstrate that the mitochondrion-localized P-subfamily PPR protein OsPPR939 is required for the splicing of nad5 introns 1, 2, and 3 in rice. Complete knockout or partial disruption of OsPPR939 function resulted in different degrees of growth retardation and pollen sterility. The dramatically reduced splicing efficiency of these introns in osppr939-4 and osppr939-5 led to reduced mitochondrial complex I abundance and activity and enhanced expression of alternative respiratory pathway genes. Complementation with OsPPR939 rescued the defective plant morphology of osppr939-4 and restored its decreased splicing efficiency of nad5 introns 1, 2, and 3. Therefore, OsPPR939 plays crucial roles in plant growth and pollen development by splicing mitochondrial nad5 introns 1, 2, and 3. More importantly, the 12th amino acid Ser in the N-terminal targeting sequence of OsPPR939 is phosphorylated by OsS6K1, and truncated OsPPR939 with a non-phosphorylatable S12A mutation in its presequence could not be imported into mitochondria, suggesting that phosphorylation of this amino acid plays an important role in the mitochondrial import of OsPPR939. To our knowledge, the 12th residue Ser on OsPPR939 is the first experimentally proven phosphorylation site in PPR proteins. Our results provide a basis for investigating the regulatory mechanism of PPR proteins at the post-translational level.

The factors influencing the unmet effects of alpha hydroxy acid peels on acne vulgaris: A single tertiary center experience.

Acne vulgaris is a common condition. Alpha hydroxy acid (AHA) peels have been previously recommended as an option for patients with intolerance to first-line treatments especially when long-term use. The safety and efficacy of AHA peels for acne have been established, but the factors influencing the ultimate effects are unknown. We recruited patients diagnosed with acne who were intolerant to or refused the first-line treatments from July 2017 to December 2019 at our hospital and retrospectively collected the medical and demographic information of patients treated with a full course of AHA peels; data collected included age, sex, treatment history, compliance status, and efficacy after treatment. The efficacy score was defined by revised scales: 2 points indicated significant improvement, 1 point indicated mild improvement, and 0 point indicated no improvement. Additionally, only efficacy scores of 2 points were classified in the satisfactory group; the others were classified in the unmet effect group. Analyses were used to evaluate the potential influencing factor(s). A total of 141 patients (120 females, 21 male) were included in the final analysis. The patients in the satisfactory group were significantly older (higher proportion aged >28 years) (47.4% vs 29.5%, P = .046) and showed better compliance (88.7% vs 54.5%, P = .001) than the patients in the unmet effect group. Multiple analyses confirmed the effects of younger age (OR 2.70, 95% CI 1.15-6.34, P = 022) and poor compliance (OR 2.74, 95% CI 1.74-4.32, P = .001) on the unmet effect. Unsatisfactory compliance and age might decrease the effects of AHA peels on acne.

Homoharringtonine Inhibits Alzheimer's Disease Progression by Reducing Neuroinflammation via STAT3 Signaling in APP/PS1 Mice.

BACKGROUND: Accumulating evidence suggests an implication of neuroinflammation in Alzheimer's disease (AD) pathogenesis. Homoharringtonine (HHT) is an antitumor reagent with anti-inflammatory activity. This study investigates whether and how HHT plays a role in disease progression in a mouse AD model. METHODS: HHT was injected into APP/PS1 mice every other day for 6 months. The effects of HHT on cognitive function were assessed by behavioral assays. ß-Amyloid accumulation was assessed by ELISA analysis of Aß40 and Aß42. Neuronal loss and synaptic function were determined by levels of NeuN, synaptophysin, and PSD95. Neuroinflammation was assessed by glial markers and pro-inflammatory cytokines. Signal transducer and activator of transcription 3 (STAT3) signaling was evaluated by phosphorylated STAT3 and SOCS3 expression. RESULTS: We found that HHT at 2 mg/kg significantly alleviated cognitive deficits in APP/PS1 mice. HHT reduced soluble and insoluble Aß40 and Aß42 accumulation and attenuated the impairments of synaptic function in the AD mouse hippocampus. Finally, HHT inhibited neuroinflammation, suppressed STAT3 activation, and increased SOCS3 expression in the APP/PS1 mouse hippocampus. CONCLUSION: Our results indicate that HHT inhibits disease progression in APP/PS1 mice by suppressing neuroinflammation through modulating the STAT3 signaling. Our findings suggest that HHT may potentially be used for preventing or slowing down AD pathogenesis and warrants further investigation.

Emergency stress management among nurses: A lesson from the COVID-19 outbreak in China-a cross-sectional study.

AIMS AND OBJECTIVES: To assess the level of stress response, self-efficacy and perceived social support status of working nurses during the outbreak of the COVID-19 and investigate potential factors affecting their stress. BACKGROUND: The stress level of clinical nurses directly affects their physical and mental health and work efficiency. DESIGN: This study was a cross-sectional investigation, which was performed following the STROBE checklist. The current study was conducted in February 2020 by selecting clinical nurses from the Zigong First People's Hospital for investigation. METHODS: At the peak of the COVID-19 outbreak in China, we assessed clinical nurses with the Stanford Acute Stress Reaction Questionnaire, the General Self-Efficacy Scale and the Perceived Social Support Scale. Specifically, the nurses were divided into three groups: (a) nurses supporting Wuhan; (b) nurses in the department of treating the COVID-19 patients in our hospital (epidemic department); and (c) nurses in the general department without the COVID-19 patients in our hospital (non-epidemic department). RESULTS: A total of 1092 clinical nurses were surveyed with 94 nurses in Wuhan, 130 nurses treating COVID-19 patients in our hospital and 868 nurses working without direct contact with diagnosed COVID-19 patients. The mean stress score of all surveyed nurses was 33.15 (SD: 25.551). There was a statistically significant difference in stress response scores between different departments. Noticeably, the nurses who went to support in Wuhan showed a weaker stress response than the nurses who stayed in our hospital (mean: 19.98 (Wuhan) vs. 32.70 (epidemic department in our hospital) vs. 34.64 (non-epidemic department in our hospital)). In addition, stress was negatively correlated with general self-efficacy and perceived social support. CONCLUSION: The present study suggested that the stress status of second-line nurse without direct contact with diagnosed COVID-19 patients was more severe than that of first-line nurses who had direct contact with COVID-19 patients. RELEVANCE TO CLINICAL PRACTICE: Our study indicated the importance of psychological status of second-line medical staff during the global pandemic.

MG53, A Novel Regulator of KChIP2 and Ito,f, Plays a Critical Role in Electrophysiological Remodeling in Cardiac Hypertrophy.

BACKGROUND: KChIP2 (K+ channel interacting protein) is the auxiliary subunit of the fast transient outward K+ current ( Ito,f) in the heart, and insufficient KChIP2 expression induces Ito,f downregulation and arrhythmogenesis in cardiac hypertrophy. Studies have shown muscle-specific mitsugumin 53 (MG53) has promiscuity of function in the context of normal and diseased heart. This study investigates the possible roles of cardiac MG53 in regulation of KChIP2 expression and Ito,f, and the arrhythmogenic potential in hypertrophy. METHODS: MG53 expression is manipulated by genetic ablation of MG53 in mice and adenoviral overexpression or knockdown of MG53 by RNA interference in cultured neonatal rat ventricular myocytes. Cardiomyocyte hypertrophy is produced by phenylephrine stimulation in neonatal rat ventricular myocytes, and pressure overload-induced mouse cardiac hypertrophy is produced by transverse aortic constriction. RESULTS: KChIP2 expression and Ito,f density are downregulated in hearts from MG53-knockout mice and MG53-knockdown neonatal rat ventricular myocytes, but upregulated in MG53-overexpressing cells. In phenylephrine-induced cardiomyocyte hypertrophy, MG53 expression is reduced with concomitant downregulation of KChIP2 and Ito,f, which can be reversed by MG53 overexpression, but exaggerated by MG53 knockdown. MG53 knockout enhances Ito,f remodeling and action potential duration prolongation and increases susceptibility to ventricular arrhythmia in mouse cardiac hypertrophy. Mechanistically, MG53 regulates NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) activity and subsequently controls KChIP2 transcription. Chromatin immunoprecipitation demonstrates NF-κB protein has interaction with KChIP2 gene. MG53 overexpression decreases, whereas MG53 knockdown increases NF-κB enrichment at the 5' regulatory region of KChIP2 gene. Normalizing NF-κB activity reverses the alterations in KChIP2 in MG53-overexpressing or knockdown cells. Coimmunoprecipitation and Western blotting assays demonstrate MG53 has physical interaction with TAK1 (transforming growth factor-b [TGFb]-activated kinase 1) and IκBα (nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha), critical components of the NF-κB pathway. CONCLUSIONS: These findings establish MG53 as a novel regulator of KChIP2 and Ito,f by modulating NF-κB activity and reveal its critical role in electrophysiological remodeling in cardiac hypertrophy.

miR-141-3p inhibits vascular smooth muscle cell proliferation and migration via regulating Keap1/Nrf2/HO-1 pathway.

miR-141-3p is proven to play a prominent role in various inflammation-related diseases. Nonetheless, little is known concerning the function of miR-141-3p in vascular smooth muscle cells (VSMCs) dysfunction and the underlying mechanism. ApoE knockdown (ApoE-/- ) C57BL/6 mice and human VSMCs were employed to establish atherosclerosis (AS) animal model and cell model, respectively. The expressions of miR-141-3p and Keap1 mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was conducted to determine inflammatory cytokines IL-6, IL-ß and TNF-α. Cell proliferation, migration and apoptosis were analyzed by BrdU assay, Transwell assay and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, respectively. Luciferase reporter assay was carried out to determine the regulatory relationship between miR-141-3p and Keap1. Additionally, Western blot was used to detect the function of miR-141-3p on the expression levels of Keap1, Nrf2 and HO-1 in VSMCs. miR-141-3p was remarkably down-regulated in both AS animal model and cell model while the expression of Keap1 was elevated. Proliferation and migration of VSMCs were suppressed after miR-141-3p mimics transfection and cell apoptosis was promoted. miR-141-3p also inhibited the expressions of IL-6, IL-ß, TNF-α and Keap1 but promoted the expressions of Nrf2 and HO-1. Moreover, the binding site between miR-141-3p and the 3'UTR of Keap1 was confirmed. miR-141-3p is down-regulated during AS, and it can alleviate VSMCs' dysfunction by targeting the Keap1/Nrf2/HO-1 axis.

Toll-like receptor 4-induced ryanodine receptor 2 oxidation and sarcoplasmic reticulum Ca2+ leakage promote cardiac contractile dysfunction in sepsis.

Studies suggest the potential role of a sarcoplasmic reticulum (SR) Ca2+ leak in cardiac contractile dysfunction in sepsis. However, direct supporting evidence is lacking, and the mechanisms underlying this SR leak are poorly understood. Here, we investigated the changes in cardiac Ca2+ handling and contraction in LPS-treated rat cardiomyocytes and a mouse model of polymicrobial sepsis produced by cecal ligation and puncture (CLP). LPS decreased the systolic Ca2+ transient and myocyte contraction as well as SR Ca2+ content. Meanwhile, LPS increased Ca2+ spark-mediated SR Ca2+ leak. Preventing the SR leak with ryanodine receptor (RyR) blocker tetracaine restored SR load and increased myocyte contraction. Similar alterations in Ca2+ handling were observed in cardiomyocytes from CLP mice. Treatment with JTV-519, an anti-SR leak drug, restored Ca2+ handling and improved cardiac function. In the LPS-treated cardiomyocytes, mitochondrial reactive oxygen species and oxidative stress in RyR2 were increased, whereas the levels of the RyR2-associated FK506-binding protein 1B (FKBP12.6) were decreased. The Toll-like receptor 4 (TLR4)-specific inhibitor TAK-242 reduced the oxidative stress in LPS-treated cells, decreased the SR leak, and normalized Ca2+ handling and myocyte contraction. Consistently, TLR4 deletion significantly improved cardiac function and corrected abnormal Ca2+ handling in the CLP mice. This study provides evidence for the critical role of the SR Ca2+ leak in the development of septic cardiomyopathy and highlights the therapeutic potential of JTV-519 by preventing SR leak. Furthermore, it reveals that TLR4 activation-induced mitochondrial reactive oxygen species production and the resulting oxidative stress in RyR2 contribute to the SR Ca2+ leak.

Baicalein triazole prevents respiratory tract infection by RSV through suppression of oxidative damage.

Baicalein, an isolate of secutellaria baicalensis is known for its anti-inflammatory activity. In the present study, 12-triazole derivatives of baicalein were synthesized and evaluated against RSV infected BEAS-2B cells in vitro and in mice model in vivo. The preventive effect of most active compound 5f against RSV infection was studied in detail. The compound 5f treatment increased IFN-ß1 expression in BEAS-2B cells infected with RSV. In BEAS-2B cells treatment with compound 5f inhibited RSV-induced secretion of interleukin-6 and -8 cytokines. It decreased RSV-induced nitric oxide & malondialdehyde production and inhibited the RSV-mediated activation of NF-κB, COX-2, Stat3 and MAPK. The p38 phosphorylation was enhanced significantly in RSV infected cells by compound 5f pre-treatment. RT-qPCR showed that compound 5f treatment of the RSV-infected mice significantly (P < 0.05) decreased viral load through reduction in the viral replication. In the mice model of RSV-infection compound 5f treatment decreased interleukin-6, -8 and tumor necrosis factor-α expression. The level of MPO, nitric oxide and malondialdehyde was also decreased significantly by compound 5f in the RSV infected mice BALF. It also reduced the infiltration of neutrophils and lymphocytes in the BALF of RVS-infected mice. In summary, compound 5f inhibits RSV-infection and prevents pulmonary airway inflammation through the activation of IFN signalling pathway.

Computer-Aided Diagnosis (CAD) of Pulmonary Nodule of Thoracic CT Image Using Transfer Learning.

Computer-aided diagnosis (CAD) has already been widely used in medical image processing. We recently make another trial to implement convolutional neural network (CNN) on the classification of pulmonary nodules of thoracic CT images. The biggest challenge in medical image classification with the help of CNN is the difficulty of acquiring enough samples, and overfitting is a common problem when there are not enough images for training. Transfer learning has been verified as reasonable in dealing with such problems with an acceptable loss value. We use the classic LeNet-5 model to classify pulmonary nodules of thoracic CT images, including benign and malignant pulmonary nodules, and different malignancies of the malignant nodules. The CT images are obtained from Lung Image Database Consortium and Image Database Resource Initiative (LIDC-IDRI) where both pulmonary nodule scanning and nodule annotations are available. These images are labeled and stored in a medical images knowledge base (KB), which is designed and implemented in our previous work. We implement the 10-folder cross validation (CV) to testify the robustness of the classification model we trained. The result demonstrates that the transfer learning of the LeNet-5 is good for classifying pulmonary nodules of thoracic CT images, and the average values of Top-1 accuracy are 97.041% and 96.685% respectively. We believe that our work is beneficial and has potential for practical diagnosis of lung nodules.

TRANSPARENT TESTA 4-mediated flavonoids negatively affect embryonic fatty acid biosynthesis in Arabidopsis.

Flavonoids are involved in many physiological processes in plants. TRANSPARENT TESTA 4 (TT4) acts at the first step of flavonoid biosynthesis, and the loss of TT4 function causes a lack of flavonoid. Flavonoid deficiency is reportedly the main cause of increased fatty acid content in pale-coloured oilseeds, but details regarding the relationship between seed flavonoids and fatty acid biosynthesis are elusive. In this work, we applied a genetic strategy combined with biochemical and cytological assays to determine the effect of seed flavonoids on the biosynthesis of fatty acids in Arabidopsis thaliana. We showed that TT4-mediated flavonoids negatively affect embryonic fatty acid biosynthesis. A crossing experiment indicated that seed flavonoid biosynthesis and the impact of this process on fatty acid biosynthesis were controlled in a maternal line-dependent manner. Loss of TT4 function activated glycolysis in seed embryos, thereby enhancing fatty acid biosynthesis, but did not improve seed mucilage production. Moreover, loss of TT4 function reduced PIN-FORMED 4 expression and subsequently increased auxin accumulation in embryos. Pharmacologically and genetically elevated auxin levels enhanced seed fatty acid biosynthesis. These results indicated that flavonoids affect fatty acid biosynthesis by carbon source reallocation via regulation of WRINKLE1 and auxin transport.

Prognostic and predictive value of plasma D-dimer levels in patients with small-cell lung cancer.

BACKGROUND: To investigate the predictive value of plasma D-dimer levels for short-term therapeutic effect and progression-free survival (PFS) in patients with small-cell lung cancer (SCLC); and to analyze the correlation between baseline plasma D-dimer levels and other clinicopathological features. The aim of the study was to investigate whether the levels of plasma D-dimer could serve as a predictive and prognostic factor in patients with SCLC. METHODS: A retrospective review of the clinicopathological data of 160 patients with pathologically confirmed SCLC, who were treated at the Department of Thoracic Oncology of Tianjin Medical University Tumor Institute and Hospital between June 2011 and June 2016, was performed. At the same time, we collected 100 patients with benign pulmonary diseases as a control group. The correlations between baseline plasma D-dimer levels and other clinical features, therapeutic effect and PFS were analyzed statistically. RESULTS: The level of plasma D-dimer in patients with SCLC was significantly higher than that of patients with benign pulmonary diseases (P = 0.001). The PFS of patients with elevated D-dimer levels before therapy were significantly shorter than that of patients with normal D-dimer levels (6.0 versus 7.5 months, P = 0.013). The patients whose plasma D-dimer level always (before and after treatment) in the normal range have the best prognosis, and continuously elevated D-dimer carried out a poor prognosis (8.0 versus 5.0 months). According to multivariate analysis, elevated D-dimer level was confirmed to be an independent prognostic factor for worse survival (P = 0.029). The level of D-dimer was associated with tumor stage, the level of neuron-specific enolase, the presence of distant metastasis, hyponatremia, and the Karnofsky performance status score; and levels decreased when therapy was effective, but increased when the disease progressed. CONCLUSIONS: High levels of baseline plasma D-dimer may indicate advanced disease stage and poor prognosis. Therefore, plasma D-dimer levels could serve as a predictive and prognostic factor in patients with SCLC.