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Thymic epithelial cells (TECs) are critical for the development and generation of functionally competent T cells. Until now, the mechanism that regulates the survival of TECs is poorly understood. In the current study, we found that Tsc1 controls the homeostasis of medullary TECs (mTECs) by inhibiting lysosomal-mediated apoptosis pathway in mice. TEC-specific deletion of Tsc1 predominately decreased the cell number of mTECs and, to a lesser content, affected the development cortical TECs. The defect of mTECs caused by Tsc1 deficiency in mice impaired thymocyte development and peripheral T cell homeostasis. Mechanistically, Tsc1 deficiency did not affect the cell proliferation of mTECs but increased the apoptosis of mTECs significantly. RNA-sequencing analysis showed that pathways involved in lysosomal biogenesis, cell metabolism, and apoptosis were remarkably elevated in Tsc1-deficient mTECs compared with their wild-type counterparts. Tsc1-deficient mTECs exhibited overproduction of reactive oxygen species and malfunction of lysosome, with lysosome membrane permeabilization and the release of cathepsin B and cathepsin L to the cytosol, which then lead to Bid cleaved into active truncated Bid and subsequently intrinsic apoptosis. Finally, we showed that the impaired development of mTECs could be partially reversed by decreasing mTORC1 activity via haploinsufficiency of Raptor. Thus, Tsc1 is essential for the homeostasis of mTECs by inhibiting lysosomal-mediated apoptosis through mTORC1-dependent pathways.

Mental fatigue is a common phenomenon with implicit and multidimensional properties. It brings dynamic changes in functional brain networks. However, the challenging problem of false positives appears when the connectivity is estimated by electroencephalography (EEG). In this article, we propose a novel framework based on spatial clustering to explore the sources of mental fatigue and functional activity changes caused by them. To suppress false positive observations, spatial clustering is implemented in brain networks. The nodes extracted by spatial clustering are registered back to the functional magnetic resonance imaging (fMRI) source space to determine the sources of mental fatigue. The wavelet entropy of EEG in a sliding window is calculated to find the temporal features of mental fatigue. Our experimental results show that the extracted nodes correspond to the fMRI sources across different subjects and different tasks. The entropy values on the extracted nodes demonstrate clearer staged decreasing changes (deactivation). Additionally, the synchronization among the extracted nodes is stronger than that among all the nodes in the deactivation stage. The initial time of the strong synchronized deactivation is consistent with the subjective fatigue time reported by the subjects themselves. It means the synchronization and deactivation correspond to the subjective feelings of fatigue. Therefore, this functional activity pattern may be caused by sources of mental fatigue. The proposed framework is useful for a wide range of prolonged functional imaging and fatigue detection studies.

In the medical image analysis, recognition of tumor in brain is very important task and it leads cancer which should be diagnosed at early stage. It is an irregular cell population in brain and for the cancer diagnosis; medical imaging techniques play an important role. The mostly used and efficient technique for the segmentation is Magnetic resonance imaging (MRI). There is huge progress the field of MRI imaging technique for accessing the brain injury and the brain anatomy exploring. The segmentation and detection of the tumor from the MRI images are done by the image processing techniques. Manual detection of brain tumor is the complex task, so the different image segmentation methods are developed for detection and segmentation of the tumor from the MRI images. The various recent brain tumor segmentation techniques are thoroughly discussed in this paper. The quantitative analysis of existing techniques and the performance evaluation is done and detailed. The paper revealed different image segmentation methods are briefly discussed. This survey article provides the detailed information of the different segmentation methods along with their merits and demerits. Effectiveness of the methods is shown in terms of the performance parameters.

Post-traumatic stress disorder (PTSD) is a serious psychiatric disorder characterized by hyper-response to environmental cues as well as the associated depressive and cognitive dysfunctions. According to the key roles of hippocampus for cognitive and emotional regulation, improving hippocampal functions, particularly hippocampal neural plasticity, is the necessary pathway to attenuate the core symptoms of PTSD. The effects of the alternative therapies such as exercise and natural compounds to reduce PTSD symptoms and promote adult hippocampal neurogenesis have been widely demonstrated. However, what is the effect of combining the exercise with traditional Chinese medical compounds remains unknown. In current study, we evaluated the effects of catalpol, which showed the pro-neurogenic effects in previous report, in regulating exercise-mediated PTSD therapeutic effects. With behavioral tests, we found that catalpol treatment promoted the effects of exercise to reduce the response of mice to dangerous cues, and simultaneously enhanced the antidepressant and cognitive protection effects. Moreover, by immunofluorescence we identified that catalpol promoted exercise-mediated hippocampal neurogenesis by enhancing the neural differentiation and mature neuronal survive. We further found that the promote effects of catalpol to exercise-induced environmental hyper-response, antidepressant effects and cognitive protective effects were all compromised by blocking neurogenesis with temozolomide (TMZ). This result indicates that hippocampal neurogenesis is prerequisite for catalpol to promote exercise-mediated brain functional improvement in PTSD model. In conclusion, our research identified the new function of natural compounds catalpol to promote the exercise-mediated brain functional changes in PTSD model, which depend on its effect promoting adult hippocampal neurogenesis.

Jie Xia,1– 4,* Lina Sun,5,* Suqin Xu,1– 4 Qiu Xiang,1– 4 Jianping Zhao,1– 4 Weining Xiong,1– 4 Yongjian Xu,1– 4 Shuyuan Chu1– 4,6 1Department of Respiratory and Critical Care Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Sciences & Technology, Wuhan, People’s Republic of China; 2Wuhan Clinical Medical Research Center for Chronic Airway Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Sciences & Technology, Wuhan, People’s Republic of China; 3Key Laboratory of Pulmonary Diseases of Health Ministry, Tongji Hospital, Tongji Medical College, Huazhong University of Sciences & Technology, Wuhan, People’s Republic of China; 4Key Site of National Clinical Research Center for Respiratory Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Sciences & Technology, Wuhan, People’s Republic of China; 5Department of Respiratory and Critical Care Medicine, Peking University Third Hospital, Beijing, People’s Republic of China; 6Laboratory of Respiratory Disease, Affiliated Hospital of Guilin Medical University, Guilin, People’s Republic of China*These authors contributed equally to this workCorrespondence: Shuyuan Chu; Jie XiaDepartment of Respiratory and Critical Care Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Sciences & Technology, Wuhan, People’s Republic of ChinaTel +8613978345180; +8627-83665520Email emilyyuanchu@163.com; xiajhzu1215@126.comPurpose: Patients with chronic obstructive pulmonary disease (COPD) would have a poor prognosis if they were not continuously managed according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines. We aim to develop a model to classify whether COPD patients have been continuously managed according to GOLD in the previous year.Methods: The Managed group were COPD patients from a prospectivecohort from November 2017 to November 2019, who have been continuously managed according to GOLD for 1 year. The Control group were COPD patients who were not continuously managed according to GOLD. They were from a retrospective cohort from October 2016 to October 2017 in the same hospitals as the Managed group. A synthetic minority over-sampling technique (SMOTE) algorithm was used to up-sample the Managed group in a training dataset. Features for classification were selected using a support vector machine recursive feature elimination (SVM-RFE) algorithm. The classification model was developed using LibSVM, and its performance was assessed on the testing dataset.Results: The final analysis included 15 subjects in the Managed group and 191 in the Control group. SVM-RFE selects nine features including smoking history, post-bronchodilator (post-)FVC before management, and those after 1-year follow-up (BMI, moderate and severe AECOPD frequency in previous 12 months, mMRC score, post-FEV1, post-FEV1%pred, post-FVC, and post-FEV1/FVC). For our model, positive predictive value is 66.7%, F1 score is 0.978, and AUC is 0.987.Conclusion: SVM classifier combined with SVM-REF feature selection algorithm could achieve good classification between COPD patients who are or are not continuously managed. This model could be applied in clinical practice to help doctors make decisions and enhance COPD patients’ compliance with standard treatment.Keywords: COPD, GOLD, continuous management, support vector machine

Chronic renal failure (CRF) is the final outcome of the development of chronic kidney disease with different causes. Although CRF is a common clinical disease, its pathogenesis remains to be improved. SBT-20 belongs to a class of cell-permeable peptides that target the inner mitochondrial membrane, reduce reactive oxygen species (ROS), normalize electron transport chain function, and ATP generation. Our experiment was to evaluate whether SBT-20 affected the oxidative stress and inflammatory process of CRF. The levels of ROS production, mitochondrial membrane potential, NF- κB-p65, TNF-α, Drp1, and mfn2 were measured before and after SBT-20 treatment. We observed that SBT-20 treatment inhibited H2O2-induced mitochondrial ROS production. SBT-20 could also restore the mitochondrial membrane potential and reduce the elevated levels of NF-κB-p65 and TNF-α in HK-2 cells. In vivo, the renal function of CRF mice recovered after treating with SBT-20, the levels of necrotic cells and inflammation decreased, and the morphology of mitochondria recovered. The results showed that SBT-20 had a protective effect on CRF by reducing oxidative stress, inflammation progression via down-regulating of NF-κB-p65, TNF-α, and Drp1 and upregulating of Mfn2. These data support SBT-20 could be used as a potential preparation for CRF.

Background Chronic obstructive pulmonary disease (COPD) is one of the most prevalent diseases worldwide. Episodes of acute exacerbations of COPD (AECOPD) are associated with disease severity and progression. Although substantial progress has been made in understanding the dynamics of AECOPD, little is known about the sputum microbiome of AECOPD in the Chinese population. Methods In this study, we characterized the sputum microbiomes from sputum specimens collected from healthy controls (n = 10), stable (n = 4), AECOPD (n = 36), and recovery (n = 18) stages by sequencing the V3-V4 region of the 16S rRNA gene with a HiSeq sequencer. Results Streptococcus was the most dominant genus among all the different types of sputum. A random forest model was developed to identify bacterial taxa that differentiate AECOPD samples from others. Most of the top predictors, except Pseudomonas, were less abundant in AECOPD samples. We also developed random forest models to differentiate subtypes of AECOPD based on blood eosinophil counts, the frequency of AECOPD, and sputum eosinophils. Bacterial taxa associated with Pasteurellaceae, Fusobacterium, Solobacterium, Haemophilus, Atopobium, Corynebacterium and Streptococcus, were enriched in the sputum microbiomes of eosinophilic AECOPD. Random forest models also demonstrate that a total of 2 bacterial OTUs were needed to differentiate frequent from non-frequent AECOPDs, and 23 OTUs were enough to accurately predict sputum-eosinophilic (sputum eosinophilic concentration ≥ 3%) AECOPD. Conclusion This study expanded our understanding of the sputum microbiome associated with different subtypes and clinical status of patients with AECOPD in a Chinese cohort, which provides insights into novel and more targeted management of the different subtypes of AECOPD.

Neonatal hypoxia can induce the persisting brain dysfunctions and subsequently result in the behavioral abnormalities in adulthood. Improving mitochondrial functions were suggested as the effective strategy for brain functional recovery. In this study, we tested the effects of physical exercise, a well-established way benefits mitochondrion, for its functions to prevent hypoxia induced adult behavioral dysfunctions and the underlying molecular mechanism. Mice was induced with hypoxia and treadmill running were then administrated until the adulthood. The treadmill running resulted in the improved behavioral performance in depressive and anxiety tests together with the enhancement of hippocampal neurogenesis. We then detected treadmill running restored the mitochondrial morphology in adult neural stem cells (NSCs) as well as the ATP production in hippocampal tissue. In addition, activity of AMPK, which playing key roles in regulating mitochondrial functions, was also elevated by treadmill running. Blockage of AMPK with selective inhibitor compound C prohibited effects of treadmill running in attenuating neonatal hypoxia induced neurogenic impairment and antidepressant behavioral deficits in adulthood. In conclusion, treadmill running could prevent neonatal hypoxia induced adult antidepressant dysfunctions and neurogenic dampening via AMPK-mediated mitochondrial regulation.

Background The small GTPase Ran is upregulated in multiple cancers and fundamental for cancer cell survival and progression, but its significance and molecular mechanisms in colorectal cancer (CRC) remain elusive. Methods Ran expression was detected in CRC cell lines and tumour tissues. In vitro and in vivo functional assays were performed to examine the effects of Ran on cell proliferation and metastasis. The pathways and effectors regulated by Ran were explored by an unbiased screening. Bioinformatics prediction and experimental validation were used to identify the miRNA regulator for Ran. Results Ran expression was frequently increased in metastatic CRC cells and tissues, especially in metastatic tissues. The upregulation of Ran correlated with poor CRC patient prognosis. Ran silencing reduced proliferation and metastasis of CRC cells both in vitro and in vivo. Ran regulated the expression of EGFR and activation of ERK and AKT signalling pathways. miR-802 was identified as an upstream regulator of Ran and miR-802 overexpression resulted in antiproliferative and antimetastatic activities. Conclusion Our study demonstrates the oncogenic roles and underlying mechanisms of Ran in CRC and the novel miR-802/Ran/EGFR regulatory axis may provide potential biomarkers for the treatment of CRC.

The impacts of androgens and glucocorticoids on spermatogenesis have intrigued scientists for decades. 11β-hydroxylase, encoded by cyp11c1, is the key enzyme involved in the synthesis of 11-ketotestosterone and cortisol, the major androgen and glucocorticoid in fish, respectively. In the present study, a Cyp11c1 antibody was produced. Western blot and immunohistochemistry showed that Cyp11c1 was predominantly expressed in the testicular Leydig cells and head kidney interrenal cells. A mutant line of cyp11c1 was established by CRISPR/Cas9. Homozygous mutation of cyp11c1 caused a sharp decrease of serum cortisol and 11-ketotestosterone, and a delay in spermatogenesis which could be rescued by exogenous 11-ketotestosterone or testosterone, but not cortisol treatment. Intriguingly, this spermatogenesis restored spontaneously, indicating compensatory effects of other androgenic steroids. In addition, loss of Cyp11c1 led to undersized testes with a smaller efferent duct and disordered spermatogenic cysts in adult males. However, a small amount of viable sperm was produced. Taken together, our results demonstrate that cyp11c1 is important for testicular development, especially for the initiation and proper progression of spermatogenesis. 11-ketotestosterone is the most efficient androgen in tilapia.

T-cell development in the thymus undergoes the process of differentiation, selective proliferation, and survival from CD4−CD8−double negative (DN) stage to CD4+CD8+double positive (DP) stage prior to the formation of CD4+helper and CD8+cytolytic T cells ready for circulation. Each developmental stage is tightly regulated by sequentially operating molecular networks, of which only limited numbers of transcription regulators have been deciphered. Here, we identified Zfp335 transcription factor as a new player in the regulatory network controlling thymocyte development in mice. We demonstrate thatZfp335intrinsically controls DN to DP transition, as T-cell-specific deficiency inZfp335leads to a substantial accumulation of DN3 along with reduction of DP, CD4+, and CD8+thymocytes. This developmental blockade at DN stage results from the impaired intracellular TCRβ (iTCRβ) expression as well as increased susceptibility to apoptosis in thymocytes. Transcriptomic and ChIP-seq analyses revealed a direct regulation of transcription factorsBcl6andRorcby Zfp335. Importantly, enhanced expression of TCRβ andBcl6/Rorcrestores the developmental defect during DN3 to DN4 transition and improves thymocytes survival, respectively. These findings identify a critical role ofZfp335in controlling T-cell development by maintaining iTCRβ expression-mediated β-selection and independently activating cell survival signaling.

Rationale It remains unknown whether long-term ozone exposure can impair lung function. Objectives To investigate the associations between long-term ozone exposure and adult lung function in China. Methods Lung function results and diagnosis of small airways dysfunction (SAD) were collected from a cross-sectional study, China Pulmonary Health Study (N=50,991). We used multivariate linear and logistic regression models to examine the associations of long-term ozone exposure with lung function parameters and SAD, respectively, adjusting for demographic characteristics, individual risk factors, and longitudinal trend. We then performed a stratification analysis by chronic obstructive pulmonary disease (COPD). Measurements and main results We observed each 1-standard deviation (SD, 4.9 ppb) increase in warm-season ozone concentrations was associated with a 14.2 mL/s [95% confidence interval (CI): 8.8, 19.6] decrease in forced expiratory flow at 75th percentile of vital capacity and a 29.5 mL/s (95% CI: 19.6, 39.5) decrease in mean forced expiratory flow between the 25th and 75th percentile of vital capacity. The odds ratio of SAD was 1.09 (95% CI: 1.06, 1.11) for a 1-SD increase in warm-season ozone concentrations. Meanwhile, we observed a significant association with a decreased ratio of expiratory volume in 1 second to forced vital capacity (FEV1/FVC) but not with FEV1 or FVC. The association estimates were greater in the COPD group than in the non-COPD group. Conclusion We found independent associations of long-term ozone exposure with impaired small airways function and higher SAD risks, while the associations with airflow obstruction were weak. COPD patients appear to be more vulnerable.

Simple Summary Colorectal cancer (CRC) is the third leading cause for cancer related death, in which metastasis exerts a pivotal role. Therefore, we aim to find out the possible mechanism underlying CRC metastasis. We found that the level of miR-125b was elevated in normal, primary CRC, and distant metastasis tissues stepwise, and high level miR-125b was positively correlated with lymph node metastasis and tumor differentiation. In vitro and in vivo assays showed miR-125b significantly promoted CRC migration and invasion. To elucidate the potential mechanism, cystic fibrosis transmembrane conductance regulator (CFTR) and cingulin (CGN) were defined as two target genes of miR-125b. On the one hand, miR-125b promoted epithelial-mesenchymal transition (EMT) and the production and secretion of urokinase plasminogen activator (uPA) by inhibiting CFTR; on the other hand, miR-125b activated Ras Homolog Family Member A (RhoA)/Rho Kinase (ROCK) signaling by repressing CGN. Therefore, we provided a potential biomarker for CRC prevention and treatment in the future. Abstract Metastasis contributes to the poor prognosis of colorectal cancer, the causative factor of which is not fully understood. Previously, we found that miR-125b (Accession number: MIMAT0000423) contributed to cetuximab resistance in colorectal cancer (CRC). In this study, we identified a novel mechanism by which miR-125b enhances metastasis by targeting cystic fibrosis transmembrane conductance regulator (CFTR) and the tight junction-associated adaptor cingulin (CGN) in CRC. We found that miR-125b expression was upregulated in primary CRC tumors and metastatic sites compared with adjacent normal tissues. Overexpression of miR-125b in CRC cells enhanced migration capacity, while knockdown of miR-125b decreased migration and invasion. RNA-sequencing (RNA-seq) and dual-luciferase reporter assays identified CFTR and CGN as the target genes of miR-125b, and the inhibitory impact of CFTR and CGN on metastasis was further verified both in vitro and in vivo. Moreover, we found that miR-125b facilitated the epithelial-mesenchymal transition (EMT) process and the expression and secretion of urokinase plasminogen activator (uPA) by targeting CFTR and enhanced the Ras Homolog Family Member A (RhoA)/Rho Kinase (ROCK) pathway activity by targeting CGN. Together, these findings suggest miR-125b as a key functional molecule in CRC and a promising biomarker for the diagnosis and treatment of CRC.

In this study, modified microwave coconut shell activated carbon (Cu Fe/MCSAC) was used to simultaneously remove NO and Hg0 in the industrial tail gas. The influences of reaction condition were investigated, such as NH3 concentration, O2 content, gas hourly space velocity (GHSV), resistance of SO2 and H2O. Under the optimal reaction conditions (600 ppm NH3, 3% O2 and 30000 h−1 GHSV), Cu Fe/MCSAC showed high NO conversion rate (86%), high Hg0 conversion rate (61%) and low N2O concentration (81 ppm) at 225 °C. NH3 was conducive to the removal of NO, but it had a competitive adsorption effect for Hg0. O2 promoted oxidation of NO and Hg0, and led to the formation of N2O. Low GHSV had the external diffusion effect, which affected the reaction rate. High GHSV could not provide enough reaction time. H2O and SO2 were not conducive to the selectivity of SCR reaction and oxidation of Hg0.

Background: The epidemiology and clinical outcome of gastroenteropancreatic signet ring cell carcinoma (SRC) were not well illustrated. We aimed to explore the long-term epidemiology and predictors affect the overall survival (OS) of patients with SRC in gastrointestinal tract and pancreas. Methods: Annual age-adjusted incidence, OS and survival trend of patients with gastroenteropancreatic SRC were evaluated in the Surveillance, Epidemiology, and End Results (SEER) database from 2000 to 2014. Multivariate Cox hazards regression model were used to identify predictive factors of the entity disease. Results: Of the 24,613 patients in the cohort, 54.0% (13,295) were male gender. The age-adjusted incidence of SRC from January 2000 to December 2014 was decreased slightly. This trend appeared across all stages, grades and all the tumor locations, excerpt for esophagus and appendix. In regard to the incidence of SRC in gastrointestinal tract and pancreas, the stomach carried a higher incidence than other sites. Different primary site, stage and grade, and age of clinical diagnosis as well as time period of diagnosis were all found to have the significant median OS by multivariable analysis. Five years’ OS of gastroenteropancreatic SRC was improving gradually between 2000–2014. In subgroups stratified by tumor stages and grades, the most pronounced improvement of survival over the same interval was observed in the early-stage and well differentiation SRC. Conclusions: SRC in a heterogeneous US population tended to carry the poor prognosis with a high proportion of distant metastasis. The poor prognosis of SRC was mainly caused by high tumor stages and poor differentiation at diagnosis.

Objective Saikosaponin c (SSc), a compound purified from the traditional Chinese herb of Radix Bupleuri was previously identified to exhibit anti-HBV replication activity. However, the mechanism through which SSc acts against HBV remains unknown. In this study, we investigated the mechanism of SSc mediated anti-HBV activity. Methods HepG2.2.15 cells were cultured at 37 ℃ in the presence of 1–40 μg/mL of SSc or DMSO as a control. The expression profile of HBV markers, cytokines, HNF1α and HNF4α were investigated by real-time quantitative PCR, Elisa, Western blot and Dot blotting. Knockdown of HNF1α or HNF4α in HepG2.2.15 cells was mediated by two small siRNAs specifically targeting HNF1α or HNF4α. Results We found that SSc stimulates IL-6 expression, leading to attenuated HNF1α and HNF4α expression, which further mediates suppression of HBV pgRNA synthesis. Knockdown of HNF1α or HNF4α in HepG2.2.15 cells by RNA interference abrogates SSc’s anti-HBV role. Moreover, SSc is effective to both wild-type and drug-resistant HBV mutants. Conclusion SSc inhibits pgRNA synthesis by targeting HNF1α and HNF4α. These results indicate that SSc acts as a promising compound for modulating pgRNA transcription in the therapeutic strategies against HBV infection. Electronic supplementary material The online version of this article (10.1007/s00011-019-01284-2) contains supplementary material, which is available to authorized users.

Rationale Our pilot study suggested that noninvasive ventilation (NIV) reduced the need for intubation compared with conventional administration of oxygen on patients with “early” stage of mild acute respiratory distress syndrome (ARDS, PaO2/FIO2 between 200 and 300). Objectives To evaluate whether early NIV can reduce the need for invasive ventilation in patients with pneumonia-induced early mild ARDS. Methods Prospective, multicenter, randomized controlled trial (RCT) of NIV compared with conventional administration of oxygen through a Venturi mask. Primary outcome included the numbers of patients who met the intubation criteria. Results Two hundred subjects were randomized to NIV (n = 102) or control (n = 98) groups from 21 centers. Baseline characteristics were similar in the two groups. In the NIV group, PaO2/FIO2 became significantly higher than in the control group at 2 h after randomization and remained stable for the first 72 h. NIV did not decrease the proportion of patients requiring intubation than in the control group (11/102 vs. 9/98, 10.8% vs. 9.2%, p = 0.706). The ICU mortality was similar in the two groups (7/102 vs. 7/98, 4.9% vs. 3.1%, p = 0.721). Multivariate analysis showed minute ventilation greater than 11 L/min at 48 h was the independent risk factor for NIV failure (OR, 1.176 [95% CI, 1.005–1.379], p = 0.043). Conclusions Treatment with NIV did not reduce the need for intubation among patients with pneumonia-induced early mild ARDS, despite the improved PaO2/FIO2 observed with NIV compared with standard oxygen therapy. High minute ventilation may predict NIV failure. Trial registration NCT01581229. Registered 19 April 2012 Electronic supplementary material The online version of this article (10.1186/s13054-019-2575-6) contains supplementary material, which is available to authorized users.

This study developed a novel method for the determination of 13 organophosphate esters (OPEs) in aqueous samples through the optimization of solvent demulsification-dispersive liquid-liquid microextraction based on solidification of floating organic drop procedure coupled with ultra-high-performance liquid chromatography-tandem mass spectrometry. The proposed method was rapid and accurate and could be used in field applications. Under the most suitable conditions, the limit of detection and limit of quantification ranged from 0.16 ng/L to 20.0 ng/L and from 0.55 ng/L to 66.7 ng/L, respectively. The enrichment factors (EFs) ranged from 30 to 46. The relative standard deviations were less than 15%. The spiked recoveries ranged between 68.2% and 97.7% in the analysis of actual aqueous samples. The proposed method was convenient, environment friendly, and time and solvent saving and could be used in field applications compared with other methods. Various concentrations and types of OPEs were detected in tap water, river water, and effluent of sewage treatment plant. Effluent samples had the highest detected levels and types of OPEs.

Fish sex could be reversed at the undifferentiated stage of gonad by administration of exogenous estrogen (E2) or blockade of endogenous estrogen synthesis with aromatase inhibitors, which is designated as primary sex reversal (PSR). Recent studies have well demonstrated that gonochoristic fish maintain their sexual plasticity after sex determination/differentiation. The differentiated ovary could be transdifferentiated into functional testis, and vice versa, the differentiated testis could be transdifferentiated into ovary. By analyzing these two secondary sex reversal (SSR) models, it was found that induction of male-to-female sex reversal initiates from dorsal (near the blood vessel) to the ventral, while induction of female-to-male sex reversal initiates from the ventral to dorsal. Down regulation of endogenous estrogen is the prerequisite for the ovarian transdifferentiation. However, exogenous estrogen alone is not sufficient for inducing differentiated testis to ovary. Administration of E2 and simultaneous blockage of androgen synthesis could induce testicular transdifferentiation. Therefore, endogenous estrogen is critical for the ovarian differentiation/maintenance and androgen is critical for testicular maintenance. Recently, genetic studies with genome editing technologies also showed that disruption of Cyp19a1a induced testicular development, indicating that cyp19a1a is the key gene essential for estrogen synthesis and ovary differentiation/maintenance. Knockout of male pathway genes or overexpression of female pathway genes could up-regulate cyp19a1a expression and increase estrogen level so as to promote ovary. Conversely, knockout of female pathway genes or overexpression of male pathway genes could down-regulate cyp19a1a expression and decrease estrogen level so as to promote testis (transgenic or knockout sex reversal, TSR). Epigenetic regulation of cyp19a1a play a critical role in natural sex reversal (NSR), but its relation with PSR, SSR and TSR needs further detailed investigations. In all, these studies further highlighted the important roles of endogenous estrogens in fish sex differentiation/maintenance.

Mucin 1 (MUC1) is a versatile oncoprotein that promotes the migration and invasion of breast cancer cells. MUC1 is up-regulated by pro-metastatic stimuli in breast cancer cells. The underlying epigenetic mechanism, however, is not fully understood. Here we report that brahma related gene 1 (BRG1), a chromatin remodeling protein, was up-regulated in highly malignant breast cancer biopsy specimens compared to those of lesser malignancy. Over-expression of BRG1 potentiated whereas depletion of BRG1 attenuated migration and invasion of MCF-7 cells. BRG1 directly bound to the MUC1 promoter and activated MUC1 transcription in a STAT1-and RelA-dependent manner. BRG1 contributed to MUC1 trans-activation by modulating histone modifications surrounding the MUC1 promoter. Specifically, BRG1 interacted with and recruited KDM3A to demethylate H3K9 on the MUC1 promoter thereby activating MUC1 transcription. Depletion of KDM3A ameliorated migration and invasion of MCF-7 cells. Altogether, our data elucidate a novel epigenetic pathway that links MUC1 transcription to breast cancer metastasis.

The emergence of antibiotic resistance has prompted the development of rapid antimicrobial susceptibility testing (AST) technologies to guide antibiotic prescription. A novel electrochemiluminescence (ECL) biosensor developed can quantitatively measure the binding between the lectin and lipopolysaccharide (LPS) on Gram-negative bacteria for bacterial determination and to characterize the antimicrobial activities of β-lactam and non-β-lactam antibiotics to normal and antibiotic-resistant bacteria. The biosensor utilizes ruthenium complex tagged concanavalin A (Ru-Con A) coated on NH2-MIL-53(Al) interface for LPS binding measurements. The decreased ECL signal obtained was directly proportional to increasing Escherichia coli (E. coli) BL21 concentrations. The sensitivity displayed logarithmic dependence in the range of (50-5.0) × 104 cells/mL, with a detection limit of 16 cells/mL. The minimum inhibitory concentration (MIC) values of antibiotics for normal E. coli BL21 were 0.02-0.2, 2-4, 0.002-0.02, and 0.2-1 mg/L for levofloxacin hydrochloride (LVX), tetracycline (TCY), imipenem (IPM), and cefpirome (CPO), respectively. The increased MIC values (8-16 and 4 mg/L for IMP and CPO, respectively) in New Delhi metallo-β-lactamase-1 expressing E. coli BL21 (NDM-1-E. coli BL21) indicated greater resistance to β-lactams in NDM-1-E. coli BL21 compared with normal E. coli BL21. Therefore, the changed ECL signal because of binding between LPS with the lectin has a relation with the type of antibiotic and bacterial strains, making the ECL biosensor promote clinical practicability and facilitate antibiotic stewardship.

Mast cell (MC) activation is associated with myocardial ischemia reperfusion injury (MIRI). Suppression of MC degranulation might be a target of anti-MIRI. This study aimed to determine whether clemastine fumarate (CLE) could attenuate MIRI by inhibiting MC degranulation. A rat ischemia and reperfusion (I/R) model was established by ligating the left anterior descending coronary artery for 30 min followed by reperfusion for 120 min. Compound 48/80 (C48/80) was used to promote MC degranulation. The protective effect of CLE by inhibiting MC degranulation on I/R injury was detected by cardiac function, 2,3,5-triphenyl tetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, arrhythmia, and myocardial enzyme detection. Inflammatory factor mRNA levels, such as TNF-α, interleukin (IL)-1β, and IL-6, were detected. Cultured RBL-2H3 mast cells were pretreated with CLE and subjected to C48/80 treatment to determine whether CLE suppressed MC degranulation. Degranulation of MCs was visualized using tryptase release, Cell Counting Kit-8 (CCK-8), and cell toluidine blue (TB) staining. RBL cells were conditionally cultured with H9C2 cells to explore whether CLE could reverse the apoptosis of cardiomyocytes induced by MC degranulation. Apoptosis of H9C2 cells was detected by CCK-8, the LDH Cytotoxicity Assay Kit (LDH), TUNEL staining, and protein expression of BAX and Bcl-2. We found that CLE pretreatment further inhibited cardiac injury manifested by decreased infarct size, histopathological changes, arrhythmias, MC degranulation, and myocardial enzyme levels, improving cardiac function compared with that in the I/R group. C48/80 combined with I/R exacerbated these changes. However, pretreatment with CLE for C48/80 combined with I/R significantly reversed these injuries. In addition, CLE pretreatment improved the vitality of RBL cells and reduced tryptase release in vitro. Similarly, the supernatant of RBL cells pretreated with CLE decreased the cytotoxicity, TUNEL-positive cell rate, and BAX expression of conditioned H9C2 cells and increased the cell vitality and expression of Bcl-2. These results suggested that pretreatment with CLE confers protection against I/R injury by inhibiting MC degranulation.

Purpose: Muscle dysfunction is considered as a sign of poor prognosis in patients with type 2 diabetes (T2D). Thus, early detection of muscle disorders is particularly important in T2D population. For free fatty acid (FFA) is known as a clinical indicator of metabolic diseases and muscle function, hence, we aimed to investigate whether FFA could serve as a biomarker for muscle function.Methods: There are totally 160 adult subjects with T2D were characterized and analyzed in this study. Muscle mass and function were measured by walking speed, grip strength and height-adjusted appendicular skeletal muscle mass (SMI). Logistic regression was applied to explore the correlation of FFA with muscle indicators. Receiver operating characteristic (ROC) curves were utilized to conduct the diagnostic value of FFA.Results: FFA was negatively correlated with SMI (r = -0.347, P = 1.0E-05), grip strength (r = -0.313, P = 7.1E-05) and walking speed (r = -0.167, P = 0.039). Notably, the relationships between FFA and SMI and walking speed remained significant even after adjusting for age, sex, body mass index (BMI) and hemoglobin A1C (HbA1c). The combination of conventional indicators including age, BMI and HbA1c provided discrimination effect of low grip strength with an AUC of 0.648, low walking speed with an AUC of 0.714. Importantly, when FFA was added to the model, the value of ROC curve was further improved, with an AUC of 0.785 for low grip strength, 0.755 for low walking speed.Conclusions: The current study demonstrated a negative correlation of FFA with muscle indicators in adult T2D patients after adjusting for HbA1c. FFA may play an important role in the pathological process of muscle dysfunction in adults with T2D.

microRNAs (miRNAs) are important components of non-coding RNAs that participate in diverse life activities by regulating gene expression at the post transcriptional level through base complementary pairing with 3'UTRs of target mRNAs. miR-133b is a member of the miR-133 family, which play important roles in muscle differentiation and tumorigenesis. Recently, miR-133b was reported to affect estrogen synthesis by targeting foxl2 in mouse, while its role in fish reproduction remains to be elucidated. In the present study, we isolated the complete sequence of miR-133b, which was highly expressed in tilapia ovary at 30 and 90 dah (days after hatching) and subsequently decreased at 120 to 150 dah by qPCR. Interestingly, only a few oogonia were remained in the antagomir-133b treated tilapia ovary, while phase I and II oocytes were observed in the ovaries of the control group. Unexpectedly, the expression of foxl2 and cyp19a1a, as well as estradiol levels in serum were increased in the treated group. Furthermore, tagln2, an important factor for oogenesis, was predicted as the target gene of miR-133b, which was confirmed by dual luciferase reporter vector experiments. miR-133b and tagln2 were co-expressed in tilapia ovaries. Taken together, miR-133b may be involved in the early oogenesis of tilapia by regulating tagln2 expression. This study enriches the understanding of miR-133b function during oogenesis and lays a foundation for further study of the regulatory network during oogenesis.

The aim of this study was to evaluate the association between the serum levels of prealbumin and sarcopenia in older adults with type 2 diabetes mellitus.This cross-sectional study included 582 older adults with type 2 diabetes mellitus. Sarcopenia was defined based on the recently updated Asian Working Group for Sarcopenia 2019 criteria. Appendicular skeletal muscle was measured by dual energy x-ray absorptiometry. Serum levels of prealbumin, hemoglobin, hemoglobin A1c, and 25-hydroxyvitamin DThe overall prevalence of sarcopenia was 9%, of which 12% for men and 6% for women. Male participants with sarcopenia had lower prealbumin levels than those without sarcopenia (213 ± 72 versus 260 ± 56 mg/L, P0.001). The proportion of men with low prealbumin level (170 mg/L) was significantly higher in individuals with sarcopenia than in those without (31% versus 6%, respectively). In a logistic regression model, after adjusting for all potential covariates, low prealbumin (odds ratio, 4.15; 95% confidence interval, 1.13-15.25; P = 0.03) was significantly associated with sarcopenia in men, but the relationship between prealbumin and sarcopenia was not found in women.Low prealbumin levels were associated with an increased risk for sarcopenia in older men with T2DM.

Background The novel noninvasive pressure-strain loop (PSL) is a reliable tool that reflects myocardial work (MW). Systolic blood pressure (SBP) is the only independent factor for MW indices. However, afterload-related reference values have not been previously reported. The aim of the present study was to establish reference values for MW parameters by wide range SBP grading. Methods We prospectively selected healthy individuals and subjects with SBP ≥ 140 mmHg at the time of study without myocardial remodeling. MW parameters were collected and the reference values achieved were grouped by SBP in 10-mmHg. Results Significant differences were noted among the SBP-groups for global work index (GWI) and global constructive work (GCW). The majority of statistical comparisons of the differences in GWI and GCW were significant at each SBP-group. With SBP ranging from 90 to 189 mmHg, the parameters GWI and GCW tended to increase linearly with afterload. Overall, the global wasted work (GWW) tended to rise as SBP was increased, but not all of the differences noted in GWW were significant for each SBP-group. Global work efficiency (GWE) remained stable across all SBP-groups, with the exception of a slight drop noted when it exceeded 160 mmHg. Conclusions The amount of MW but not the work efficiency varied greatly according to the different afterload. This finding cannot be ignored during clinical research or diagnosis and afterload-related reference values are required to make a reasonable judgment on the myocardial function.

The incidence of chronic kidney disease (CKD) complicated with cardiovascular disease (CVD) is increasing day by day. CVD complication is a significant risk factor for the progression and poor prognosis of CKD. However, in terms of clinical diagnosis, treatment, and pathophysiological mechanism, there are still many things that are not clear about CKD combined with CVD. In this article, we observed the abnormal expression of long non-coding RNA ZFAS1 (lncRNA ZFAS1) in the heart and renal tissues of CKD mice, which may a target for nephropathy with CVD. We observed that exosomes containing lncRNA ZFAS1 induced fibrosis in the heart via the wnt4/β-catenin signal pathway. Luciferase assay reported that lncRNA ZFAS1 could bind with microRNA-4711-5p (miR-4711-5p), and wnt4 was a target of miR-4711-5p. In conclusion, lncRNA ZFAS1 from HCM could promote cardiac fibrosis through WNT/β-catenin signal via exosome pathways, which provided an underlying mechanism in the cardiac fibrosis from CKD.

Perioperative neurocognitive disorders (PND) is a common postoperative neurological complication. Neuroinflammation is a major cause that leads to PND. Autophagy, an intracellular process of lysosomal degradation, plays an important role in the development and maintenance of nervous system. PK11195 is a classic translocator protein (TSPO) ligand, which can improve the cognitive function of rats. In this study, we evaluate the protective effect of PK11195 on the learning and memory of rats. A rat model of lipopolysaccharide (LPS)-induced cognitive dysfunction was established by intraperitoneal injection of LPS. Morris Water Maze (MWM), Western blot, qRT-PCR, confocal microscopy and transmission electron microscopy (TEM) were used to study the role of TSPO-specific ligand PK11195 in LPS-activated mitochondrial autophagy in rat hippocampus. We found that PK11195 ameliorated LPS-induced learning and memory impairment, as indicated by decreased escape latencies, swimming distances and increased target quadrant platform crossing times and swimming times during MWM tests. TSPO, ATG7, ATG5, LC3B and p62 protein and mRNA expression increased in the hippocampus of PND model rats. The hippocampal microglia of PND model rats also have severe mitochondrial damage, and a large number of autophagosomes and phagocytic vesicles can be seen. PK11195 pretreatment significantly decreased the expression of TSPO, ATG7, ATG5, LC3B and p62 protein and mRNA, as well as mitochondrial damage. These findings suggested that PK11195 may alleviate the damage of LPS-induced cognitive dysfunction of rats by inhibiting microglia activation and autophagy.

Background Pulmonary infections caused by non-diphtheriae corynebacteria are increasing. However, rapid identification of Corynebacterium species poses a challenge due to the low genetic variation within the genus. Methods Three reference strains and 99 clinical isolates were used in this study. A qPCR followed by high-resolution melting (HRM) targeting ssrA was performed to simultaneously identify C. striatum, C. propinquum and C. simulans. To further evaluate this assay’s performance, 88 clinical sputum samples were tested by HRM and the detection results were compared with those of the traditional culture method and multiple cross-displacement amplification (MCDA) assay. Results The melting curve produced by a pair of universal primers generated species-specific HRM curve profiles and could distinguish the three target species from other related bacteria. The limit of detection of HRM assay for DNA from the three purified Corynebacterium species was 100 fg. Compared with the culture method, HRM detected 22 additional positive specimens, representing a 23.9% relative increase in detection rate. The HRM assay had 98.4% (95% confidence interval [CI], 90.5–99.9%) sensitivity and 100% (95% CI, 82.8–100%) specificity. Additionally, 95.5% concordance between HRM and MCDA (κ = 0.89 [95% CI, 0.79–0.99]) was noted. Conclusions The HRM assay was a simple, rapid, sensitive, and specific diagnostic tool for detecting C. striatum, C. propinquum, and C. simulans, with the potential to contribute to early diagnosis, epidemiological surveillance, and rapid response to outbreak.

Purpose To investigate the effect of age on wound healing after small incision lenticule extraction (SMILE) and the underlying metabolomic mechanisms. Methods This prospective study was conducted on 216 patients in four groups: the 18–20 (n = 38, Group I), 21–30 (n = 84, Group Ⅱ), 31–40 (n = 58, Group Ⅲ), and 41–50 (n = 36, Group IV) age groups. The density of corneal epithelial wing cells, basal cells, corneal stromal cells, endothelial cells and corneal nerves were examined with a laser confocal microscope (HRT III-RCM) before and 1 month, 3 month, 6 month and 1 year after SMILE. The central nerve fiber length (CNFL), the central corneal nerve fibre density (CNFD), and the central corneal nerve branch density (CNBD) were analyzed by Nero J. The corneal stroma lenticules were obtained from SMILE to analyze metabolites by high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-QTOF-MS). Results The density of corneal wing epithelial cells and basal epithelial cells have no significant difference among the four groups. The CNFL was 21.90 ± 1.68 mm/mm2 in Group Ⅰ and 21.63 ± 2.09 mm/mm2 in Group Ⅱ after 1 year of SMILE, which represented a return to the preoperative level, whereas the CNFL of Group Ⅲ (19.40 ± 0.98 mm/mm2) and Group Ⅳ (18.94 ± 0.72 mm/mm2) were lower than that preoperation (P ˂0.01). CNFL repair had a negative correlation with age after surgery (Pearson's R = −0.572, P ˂0.01). The CNFD and the CNBD showed the same trend with the CNFL (Pearson's R = −0.602 and −0.531, P ˂0.05). Through screening the significantly different metabolites between the 18–30 age group (including Group I and Group Ⅱ) and other two groups, 6 common remarkably different metabolites were identified. Meanwhile, 5 unique different metabolites were identified only between the 18–30 age group and the 31–40 age group. Six unique different metabolites were identified only between the 18–30 age group and the 41–50 age group. Conclusion Corneal nerve repair after SMILE was significantly affected by age. The identified age-associated differences in metabolites were mainly related to inflammation, oxidation, nerve protection and regeneration.

Objective. To observe the characteristics of ankle pain and ankle joint muscle by microscope. Methods. In a sports university, 15 athletes above grade 2 were randomly selected as the experimental group, and 15 nonathletes were randomly selected as the control group. The experiment mainly included foot shape test, standard scaphoid height test, ankle range of motion test, ankle muscle emg test, and other experimental procedures. Medical microscopic image processing is a new technology developed in the past thirty years, which has brought great progress for mankind to understand and transform nature. Among them, the image processing and recognition of tumor cell microscopic images are one of the research focuses on the use of computers to process and recognize medical images. Results. In the test of ankle range of motion, when the angular velocity was the same as 60°/s, compared with the control group, the difference value of bilateral flexor peak moment in the experimental group was large, and the difference between the two groups was significant ( P < 0.05 ), with statistical significance. As the angular velocity dropped from 240°/s to 60°/s, 30 members of the experimental group and control group also experienced a decrease in the bilateral ankle isokyclic muscles, reflecting their lack of ankle strength. On the other hand, the muscle strength of the ankle joint in the experimental group was relatively small, and the difference between the two groups was significant ( P < 0.05 ). During the exercise, some members of the experimental group suffered from ankle pain, which resulted in insufficient strength of the muscles of the ankle joint, resulting in the interruption of the experiment. In the emg test of ankle muscles, the effective discharge values of preexcitation current of tibial anterior muscle before and after exercise were 104.6 ± 26.5 and 129.2 ± 38.1 , respectively, with significant difference and statistical significance. In the foot morphology test and the standard scaphoid height test, the difference between the two groups was not significant ( P > 0.05 ), and there was no statistical significance. Conclusion. Microscope based on athlete’s foot and ankle pain and ankle muscle characteristics to improve the accuracy of the observation, with the help of a microscope, you can see the details of a doctor are invisible to the naked eye and can record the relevant data in time in order to observe the late, for athlete’s foot and ankle pain relief and enhanced ankle muscles provide data support.

Sphingolipids metabolism is an important cell process and plays critical roles in asthma. However, the involvement of sphingolipids in the pathogenesis of asthma and its subtypes is unknown. The present study aimed to determine the role of sphingolipids in asthma and its subtypes. Clinical data from 51 asthma patients and 9 healthy individuals were collected and serum samples were performed to analyze the levels of serum sphingolipids by liquid chromatography-mass spectrometry-based targeted metabolomics. Results showed that the levels of sphingomyelin (SM) including SM34:2, SM38:1, and SM40:1 were significantly decreased in asthmatic patients compared to healthy controls. Moreover, serum SM levels were obviously decreased in the blood noneosinophilic asthma (bNEA) group compared with blood eosinophilic asthma group. Similar tendencies of serum SM level changes were observed in the early-onset group compared with late-onset group. Correlation analysis revealed that SM 40:1 was negatively related to sputum IL-17A (r = −0.621, P = 0.042). The present study presented that the SM may be a protective factor of asthma and contributes to the mechanism of asthma, especially bNEA. SM may be a potential biomarker and therapeutic target in asthma.

Neuroinflammation related to microglial activation plays an important role in neurodegenerative diseases. Translocator protein 18 kDa (TSPO), a biomarker of reactive gliosis, its ligands can reduce neuroinflammation and can be used to treat neurodegenerative diseases. Therefore, we explored whether TSPO ligands exert an anti-inflammatory effect by affecting the nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome, thereby inhibiting the release of inflammatory cytokines in microglial cells. In the present study, BV-2 cells were exposed to lipopolysaccharide (LPS) for 6 h to induce an inflammatory response. We found that the levels of reactive oxygen species (ROS), NLRP3 inflammasome, interleukin-1β (IL-1β), and interleukin-18 (IL-18) were significantly increased. However, pretreatment with TSPO ligands inhibited BV-2 microglial and NLRP3 inflammasome activation and significantly reduced the levels of ROS, IL-1β, and IL-18. Furthermore, a combination of LPS and ATP was used to activate the NLRP3 inflammasome. Both pretreatment and post-treatment with TSPO ligand can downregulate the activation of NLRP3 inflammasome and IL-1β expression. Finally, we found that TSPO was involved in the regulation of NLRP3 inflammasome with TSPO ligands treatment in TSPO knockdown BV2 cells. Collectively, these results indicate that TSPO ligands are promising targets to control microglial reactivity and neuroinflammatory diseases.