Autophagy-mediated degradation of integumentary tapetum is critical for embryo pattern formation.

Autophagy modulates the degradation and recycling of intracellular materials and contributes to male gametophyte development and male fertility in plants. However, whether autophagy participates in seed development remains largely unknown. Here, we demonstrate that autophagy is crucial for timely programmed cell death (PCD) in the integumentary tapetum, the counterpart of anther tapetum, influencing embryo pattern formation and seed viability. Inhibition of autophagy resulted in delayed PCD of the integumentary tapetum and defects in embryo patterning. Cell-type-specific restoration of autophagic activities revealed that the integumentary tapetum plays a non-autonomous role in embryo patterning. Furthermore, high-throughput, comprehensive lipidomic analyzes uncovered an unexpected seed-developmental-stage-dependent role of autophagy in seed lipid metabolism: it contributes to triacylglycerol degradation before fertilization and to triacylglycerol biosynthesis after fertilization. This study highlights the critical role of autophagy in regulating timely integumentary tapetum PCD and reveals its significance in seed lipid metabolism and viability.

Effects of UGT1A, CYP2C9/19 and ABAT polymorphisms on plasma concentration of valproic acid in Chinese epilepsy patients.

Aim: To evaluate the association between genetic polymorphisms and plasma concentration-to-dose ratio of valproic acid (CDRV) in Chinese epileptic patients. Methods: A total of 46 epileptic patients treated with valproic acid therapy were enrolled. 18 SNPs in nine genes related to valproic acid were directly sequenced with Sanger methods. Results: Patients carrying UGT1A6 heterozygous genotypes had significantly lower CDRV than those carrying the wild-type genotypes. In contrast, patients with the homozygote genotypes of CYP2C9 and ABAT had higher CDRV than those with the wild-type genotypes and patients with the heterozygous genotypes of CYP2C19 had higher CDRV. Conclusion: Detection of genetic polymorphism in these genes might facilitate an appropriate dose of valproic acid for epileptic patients. Further studies with larger cohorts are necessary to underpin these observations.

Synthesis of Molecularly Imprinted Polymers Based on a New Monomer "2-(4-Vinylphenyl) Quinoline-4-Carboxylic Acid" for the Selective Solid-Phase Extraction of Lamotrigine.

A new molecularly imprinted polymers (MIPs) have been prepared for the high selective extraction of lamotrigine (LTG), a widely used antiepileptic drug, in human serum. The MIPs were polymerized by bulk polymerization using our synthesized compound, 2-(4-vinylphenyl) quinolin-4-carboxylic acid, as functional monomer, which achieved better adsorption specificity than universal MIPs. Then, the molecularly imprinted solid phase extraction (MISPE) based on this material was coupled with high-performance liquid chromatography (HPLC) for the detection of LTG in human serum. The results of method validation showed that the developed method presented a good precision and accuracy, and the linearity was in the range of 1.50-40.00 mg/mL with the limit of quantitation (LOQ) at 0.20 mg/mL. The recovery ranged from 80.8% to 83.8% with RSD ranges from 5.5% to 11.1%. The validated method was successfully used to determine the concentration of LTG in human simulate serum samples.

Clinicopathological Features of Sessile Serrated Polyps in China: A Retrospective Study of a Tertiary Hospital.

BACKGROUND: Serrated polyps have been recognized as the important premalignant lesions. In this study, we aimed to analyze the clinicopathological features of sessile serrated polyps and determine the association between sessile serrated polyps and synchronous advanced adenomas. METHODS: Consecutive patients undergoing diagnostic or therapeutic colonoscopies (including 156 681 diagnostic colonoscopies) from 2011 to 2019 were included. RESULTS: A total of 958 patients, including 699 (73%) males, were detected with at least 1 sessile serrated polyp, and 65.9% (n = 658) of sessile serrated polyps were located in the distal colon. Advanced serrated lesions accounted for 9.1% (n = 91) of all the sessile serrated polyp (n = 999). The types of SSP included flat type (953/999, 95.4%) and sub-pedunculated or pedunculated type (46/999, 4.6%). Meanwhile, there was no obvious evidence supporting the association between advanced adenomas and characteristics of advanced serrated lesions or sessile serrated polyps. CONCLUSION: Sessile serrated polyps seem to be more frequently seen in the distal colon of men in this study. However, more evidence is required to confirm the actual distribution of sessile serrated polyp in colon among Chinese people. There is still much room for improvement of sessile serrated polyp detection rate, and more importance should be attached to sessile serrated polyp both for pathologists and endoscopists.

A simple and easy non-derivatization gas chromatography-mass spectrometry method for simultaneous quantification of valproic acid, gabapentin, pregabalin, and vigabatrin in human plasma.

Immunoassays are currently not available in commercial kits for the quantification of valproic acid, vigabatrin, pregabalin, and gabapentin, which also cannot suffer the limitations of interferences of substances with similar structures. Chromatography is a good alternative to immunoassay. In this study, a simple and robust non-derivatization gas chromatography-mass spectrometry method for simultaneous determination of the above four drugs in human plasma was developed and validated for therapeutic drug monitoring purposes. This method employed benzoic acid as the internal standard with hydrochloric acid for plasma acidification and ACN for precipitate protein. The supernatant was directly injected into gas chromatography-mass spectrometry for analysis. Good linearity was obtained with linear correlation coefficients of the four analytes of 0.9988-0.9996. Extraction recoveries of valproic acid, vigabatrin, pregabalin, and gabapentin were respectively in the ranges of 91.3%-94.5%, 90.0%-90.9%, 90.0%-92.1%, and 88.0%-92.2% with the relative standard deviation values less than 12.6%. Intra- and inter-batch precision and accuracy, and stability assays were all acceptable. Taken together, the novel method developed in this study provided easy plasma pretreatment, good extraction yield, and high chromatographic resolution, which has been successfully validated through the quantification of valproic acid in the plasma of 46 patients with epilepsy.

Diagnostic accuracy of endoscopic ultrasound with artificial intelligence for gastrointestinal stromal tumors: A meta-analysis.

OBJECTIVES: Gastrointestinal stromal tumors (GISTs) are thought to have a malignant potential. However, utilizing endoscopic ultrasound (EUS) to differentiate GISTs from other types of subepithelial lesions (SELs) remains challenging. Artificial intelligence (AI)-based diagnostic systems for EUS have been reported to have a promising performance, although the results of the previous studies remain controversial. In this meta-analysis, we aimed to assess the diagnostic accuracy of AI-based EUS in distinguishing GISTs from other SELs. METHODS: A literature search was conducted on MEDLINE and EMBASE databases to identify relevant articles. The sensitivity, specificity, and area under the summary receiver operating characteristic curve (AUROC) of eligible studies were analyzed. RESULTS: Seven studies were eligible for the final analysis. The combined sensitivity and specificity of AI-based EUS were 0.93 (95% confidence interval [CI] 0.88-0.96) and 0.78 (95% CI 0.67-0.87), respectively. The overall diagnostic odds ratio of AI-based EUS for GISTs was 36.74 (95% CI 17.69-76.30) with an AUROC of 0.94. CONCLUSIONS: AI-based EUS showed high diagnostic ability and might help better differentiate GISTs from other SELs. More prospective studies on the diagnosis of GISTs using AI-based EUS are warranted in clinical setting.

Development and evaluation of a simple and easy high-performance liquid chromatography-ultraviolet system simultaneously suitable for determination of 24 anti-epileptic drugs in plasma.

We aim to establish a simple and easy high-performance liquid chromatography system coupled with an ultraviolet detector suitable for simultaneous determination of 24 antiepileptic drugs in human plasma. Optimized chromatographic separation was performed on a ZORBAX Eclipse Plus-C18 (4.6 × 150 mm2 , 3.5 µm) column with acetonitrile and 5 mM potassium dihydrogen phosphate water solution as mobile phase. Note that, 24 antiepileptic drugs were divided into three groups and eluted with different gradient procedures, respectively. The column temperature was maintained at 35°C and the detection wavelength was set at 210 nm. Plasma was processed with ethyl acetate or acetonitrile. The calibration curves of 24 antiepileptic drugs demonstrated good linearity within the test range (r > 0.996). The intra- and inter-batch precision and accuracy were all less than 15%, while extraction recoveries were in the range of 74.57-90.89% with the relative standard deviation values less than 15%. The validated methods have been successfully applied to determination of some antiepileptic drugs in rat or patient plasma. Those results indicated that the developed methods were simple and easy, and could be suitable for the determination of 24 antiepileptic drugs in plasma just by changing the gradient elution procedures of mobile phase.

Development of magnetic molecularly imprinted polymers with double templates for the rapid and selective determination of carbamazepine and lamotrigine in serum.

A dual-template magnetic molecularly imprinted polymer (Dt-MMIP) with a specific recognition capability for carbamazepine (CBZ) and lamotrigine (LTG) was synthesized using methacrylic acid as a functional monomer, and ethylene glycol dimethylmethacrylate as a cross-linking agent. A magnetic non-molecularly imprinted polymer without templates (MNIP) was also prepared using the same procedure. The prepared polymers were characterized using scanning electron microscopy, Fourier-transform infrared spectroscopy and adsorption experiments. Results indicated that both Dt-MMIPs and MNIPs were microspherical nanoparticles, and the surface of the Dt-MMIP was rougher than that of the MNIP. In addition, the prepared Dt-MMIPs possessed a higher adsorption capacity and better selectivity for CBZ and LTG than the MNIPs. The maximum static adsorption capacities of Dt-MMIP for CBZ and LTG were 249.5 and 647.9 µg g-1, respectively, whereas those of MNIP were 75.8 and 379.8 µg g-1, respectively. The obtained Dt-MMIPs were applied as a magnetic solid-phase extraction sorbent for the rapid and selective extraction of CBZ and LTG in rat serum samples, and determination was performed by high-performance liquid chromatography with UV detection (HPLC-UV). The developed method of dispersive SPE based on Dt-MMIPs coupled to HPLC-UV has good rapidity and selectivity, and application prospects in serum.

Analysis of plasma free amino acids in diabetic rat and the intervention of Ginkgo biloba leaves extract using hydrophilic interaction liquid chromatography coupled with tandem mass-spectrometry.

Amino acids (AAs) are important metabolites that are related with diabetes. However, their roles in the initiation and development of diabetes mellitus (DM), especially in the treatment of Ginkgo biloba leaves extract (GBE) have not been fully explored. Thus, we investigated the roles that AAs played in the progression and GBE supplementation of DM rat induced by streptozotocin. The rats were randomly divided into a normal control group treated with drug-free solution, a normal control group treated with GBE, a DM group treated with drug-free solution, and DM group treated with GBE; and maintained on this protocol for 9 weeks. Rat plasma was collected from the sixth week to the ninth week and then analyzed with the optimized hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry method. A total of 17 AAs with differential levels were monitored to indicate dysfunction of AAs metabolism to confirm the occurrence and development of DM. Treatment with GBE partially reversed the changes seen in seven AAs including leucine, isoleucine, tyrosine, glutamic acid, asparagines, lysine and alanine in DM rats, indicating that GBE could prevent the occurrence and development of DM by acting on AAs metabolism. The improvement of those AAs metabolism disorders may play a considerable role in the treatment of GBE on the occurrence and development of DM. Those findings potentially promote the understanding of the pathogenic progression of DM and reveal the therapeutic mechanism of GBE against DM.

[Puerarin Inhibits the Proliferation,Invasion,and Migration of Non-small Cell Lung Cancer Cells through Regulating miR-490/Denticleless E3 Ubiquitin Protein Ligase].

Objective To explore the mechanism of puerarin inhibiting the proliferation,invasion,and migration of non-small cell lung cancer cells. Methods A549 cells were cultured and treated with different concentrations of puerarin.The inhibition rate (IR) on cell proliferation was detected by CCK-8,and qRT-PCR was performed to detect the mRNA levels of miR-490 and denticleless E3 ubiquitin protein ligase(DTL).Double luciferase reporter assay was employed to identify the targets of miR-490 and DTL based on the establishment of NC mimic group,miR-490 mimic group,NC inhibitor group,and miR-490 inhibitor group.The cells treated by 20 µmol/L puerarin were classified into six groups:DMSO,puerarin,puerarin+NC inhibitor,puerarin+miR-490 inhibitor,puerarin+miR-490 inhibitor+Si-NC,and puerarin+miR-490 inhibitor+Si-DTL.Transwell was used to detect cell migration and invasion.Western blotting was performed to detect the protein levels of epithelial-mesenchymal transition-related markers E-cadherin,N-cadherin,and Vimentin. Results With the increase in puerarin concentration,the IR gradually elevated (F=105.375,P<0.001),miR-490 expression gradually increased (F=32.919,P<0.001),and DTL expression gradually decreased (F=116.120,P<0.001).Compared with NC mimic group,miR-490 mimic group had decreased luciferase activity (t=7.762,P=0.016),raised miR-490 mRNA level (t=13.319,P<0.001),and declined DTL mRNA level (t=7.415,P=0.002).Compared with those in NC inhibitor group,miR-490 demonstrated decreased mRNA level (t=9.523,P=0.001) and DTL presented increased mRNA level (t=11.305,P<0.001) in miR-490 inhibitor group.Western blotting showed that the protein level of DTL was higher in NC mimic group (t=7.953,P=0.001) than in miR-490 mimic group and higher in miR-490 inhibitor group than in NC inhibitor group (t=10.552,P<0.001).Compared with DMSO group,puerarin group showed up-regulated mRNA level of miR-490 (t=10.255,P=0.001) while down-regulated mRNA level of DTL (t=6.682,P=0.003).Compared with those in puerarin+NC inhibitor group,the mRNA level of miR-490 declined (t=10.995,P<0.001) while that of DTL raised (t=12.478,P<0.001) in puerarin+miR-490 inhibitor group.The mRNA level of miR-490 had no significant difference between puerarin+miR-490 inhibitor+Si-NC group and puerarin+miR-490 inhibitor+Si-DTL group (t=1.081,P=0.341),and that of DTL was lower in the latter group (t=14.321,P<0.001).The protein level of DTL was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (t=11.423,P<0.001),and lower in puerarin+miR-490 inhibitor+Si-DTL group than in puerarin+miR-490 inhibitor+Si-NC group (t=12.080,P<0.001).Compared with DMSO group,puerarin group showed inhibited cell proliferation (F=129.27,P<0.001).The activity of cell proliferation was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (F=75.12,P<0.001),and higher in puerarin+miR-490 inhibitor+Si-NC group than in puerarin+miR-490 inhibitor+Si-DTL group (F=52.59,P<0.001).Compared with DMSO group,puerarin group had suppressed cell migration (t=8.963,P=0.001).The cell migration ability was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (t=12.117,P<0.001) and higher in puerarin+miR-490 inhibitor+Si-NC group than in puerarin+miR-490 inhibitor+Si-DTL group (t=12.934,P<0.001).Puerarin group showed weakened cell invasion ability compared with DMSO group (t=4.710,P=0.009).The cell invasion ability was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (t=13.264,P<0.001) and lower in puerarin+miR-490 inhibitor+Si-DTL group than in puerarin+miR-490 inhibitor+Si-NC group (t=13.476,P<0.001).Compared with DMSO group,puerarin group showed up-regulated protein level of E-cadherin (t=7.137,P=0.002) while down-regulated protein levels of N-cadherin (t=8.828,P=0.001) and vimentin (t=6.594,P=0.003).Compared with those in puerarin+NC inhibitor group,the protein level of E-cadherin (t=12.376,P<0.001) decreased while those of N-cadherin (t=13.436,P<0.001) and vimentin (t=11.467,P<0.001) increased in puerarin+miR-490 inhibitor group.Compared with puerarin+miR-490 inhibitor+Si-NC group,puerarin+miR-490 inhibitor+Si-DTL group up-regulated the protein level of E-cadherin (t=13.081,P<0.001) while down-regulated the protein levels of N-cadherin (t=10.835,P<0.001) and vimentin (t=11.862,P<0.001). Conclusion Puerarin could inhibit the proliferation,invasion,and migration of non-small cell lung cancer cells by up-regulating miR-490 and down-regulating DTL.

Chemical Antioxidant Quality Markers of Chrysanthemum morifolium Using a Spectrum-Effect Approach.

Traditionally, the quality evaluation of Chrysanthemum morifolium (CM) cv. (Juhua) attributes its habitats and processing methods, however, this strategy of neglecting bioactive ingredients usually results in deviation of quality evaluation. This study aims to explore the quality marker (Q-marker) based on spectrum-effect relationship and quality control strategy of CMs. The chromatographic fingerprint of 30 flower head samples of CMs from five different habitats including Hang-baiju, Gongju, Huaiju, Taiju and Boju were constructed by high performance liquid chromatography and analyzed through chemometrics methods such as similarity analysis (SA), cluster analysis (CA) and principal component analysis (PCA). The common peaks were quantified by external standard method and relative correction factor method. The in-vitro radical scavenging capacity assays of DPPH·, ·OH and ABTS were carried out. The Q-marker was explored by the correlation analysis between the contents of common peaks and in-vitro radical scavenging capacity, and then used to evaluate the quality of 30 flower head samples of CMs. A total of eight common peaks were appointed in 30 flower head samples of CMs, and their similarities ranged from 0.640 to 0.956. CA results showed that 30 flower head samples of CMs could be divided into five categories with reference to the Euclidean distance of 5. PCA results showed that common peaks played a major role in differential contribution of CMs. The quantification of common peaks hinted that their contents possessed significant variation whether for different accessions or the same accessions of CMs. The correlation analysis showed that chlorogenic acid, 3,5-O-dicaffeoylquinic acid, unknown peak 1, 4,5-O-dicaffeoylquinic acid and kaempferol-3-O-rutinoside could be used as the Q-markers for the quality evaluation of 30 flower head samples of commercially available CMs. The analysis strategy that combines chromatographic fingerprint analysis, multiple ingredients quantification, in-vitro chemical anti-oxidant activity evaluation and spectrum-effect relationship analysis clarified the therapeutic material basis and discovered the Q-markers, which possibly offers a more comprehensive quality assessment of CMs.

Semen microbiota in normal and leukocytospermic males.

Large numbers of microbes can be present in seminal fluid, and there are differences in the semen microbiota between normal and abnormal semen samples. To evaluate the semen microbiota in patients with leukocytospermia, 87 seminal fluid samples, including 33 samples with a normal seminal leukocyte count and 54 samples with leukocytospermia, were obtained for a cross-sectional analysis. Twenty samples with a normal seminal leukocyte count had normal sperm parameters (Control group), and 13 samples with a normal seminal leukocyte count were from asthenozoospermia patients (Ast group). However, 32 samples with leukocytospermia were from asthenozoospermia patients (LA group), and only 22 samples with leukocytospermia had normal sperm parameters (Leu group). The 16S ribosomal RNA (rRNA) gene sequencing method was used to sequence the microbiota in the seminal fluid, and multiple bioinformatics methods were utilized to analyze the data. Finally, the results showed that the worse sperm parameters were observed in the leukocytospermia-related groups. Semen microbiota analysis found that there was increased alpha diversity in the leukocytospermia-related groups. Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes were the primary phyla in the seminal fluid. Two microbiota profiles, namely, Lactobacillus-enriched and Streptococcus-enriched groups, were identified in this study. The majority of the samples in the groups with a normal seminal leukocyte count could be categorized as Lactobacillus-enriched, whereas the majority of the leukocytospermia samples could be categorized as Streptococcus-enriched. Our study indicated that males with leukocytospermia have worse sperm parameters and a different semen microbiota composition compared to males with a normal seminal leukocyte count.

Bibliometrics and visualization analysis of EwE model based on CiteSpace.

To explore the historical evolution and current status of the EwE (Ecopath with Ecosim) modelling research, the core dataset and extended dataset were collected by topic retrieval and citation indexing methods from the "Web of Science" from 1984 to 2020. The bibliometric analysis and mapping knowledge were performed by CiteSpace software, focusing on literature distribution, research forces, research theme, and hotspot evolution. The results showed that the annual publications in the EwE model researches were increasing, covering multi-disciplinary fields. Christensen, Walters, and Pauly were representative scholars with an important role in model development and relevant international cooperation. In the early stage, EwE model was usually applied to solve ecosystem problems, including spatial-temporal dynamic of structure and function, and the ecosystem effects of fisheries. Currently, marine resource management, ecosystem modelling, marine protected areas and ecosystem indicators had become the key themes. The research hotspots shifted from model development and food web structure to ecosystem forecasting and resource management, which would provide scientific evidence for ecosystem-based aquatic resource management and the construction of protected area in marine.

The role of cytology in endobronchial ultrasound-guided transbronchial needle aspiration: A study of 813 cases focusing on diagnostic yield, an analysis of misdiagnosed cases and diagnostic accordance rate of cytological subtyping.

BACKGROUND: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a minimally invasive technique for cytological and histological diagnosis. The objective of this study was to explore the role of cytological diagnosis in EBUS-TBNAs. METHODS: Eight hundred and thirteen consecutive cases performed EBUS-TBNA with both cytological and histological diagnoses were retrospectively reviewed. All patients were followed up for clinical data. RESULTS: Before immunohistochemical examination, the cytological sensitivity, specificity, and diagnostic accuracy of EBUS-TBNAs were 92.9% (421/453), 98.9% (348/352), 95.5% (769/805), respectively. After immunohistochemical examination, the sensitivity, specificity, and diagnostic accuracy were 93.0% (423/455), 99.4% (348/350), 95.8% (771/805), respectively. The majority of false-negative were cases whose cytological diagnosis was "atypical" or the cytological diagnosis suggested "inadequate." "Neoplastic" were also prone to false-negative cytology. The diagnostic accordance rate of cytological subtyping was 90.3% for squamous-cell carcinoma, 99.2% for adenocarcinoma, and 98.1% for small-cell carcinoma before immunohistochemical examination, and became 85.9%, 98.5%, and 98.2% after immunohistochemical examination, respectively. CONCLUSION: Cytological diagnosis in EBUS-TBNAs had a good sensitivity and high specificity. The sensitivity and specificity of cytological diagnosis were proved to be higher after the immunohistochemical examination. At the same time, cytology had high accordance rate in subtype diagnosis. False-negative results occurred more commonly in cases whose cytological diagnosis was "atypical" or the cytological diagnosis suggested "inadequate" or the corresponding histological diagnosis was "Neoplastic."

The value of BAP1 immunocytochemistry and CDKN2A fluorescence in situ hybridization in diagnosis of serous effusion malignant mesothelioma and an analysis of the association between degree of cell atypia and the results of two auxiliary methods.

Malignant mesothelioma (MM) is a rare, highly aggressive tumor. The first symptom of MM is mostly serous effusion, and cytology can be used in diagnosis based on effusion, providing patients with an earlier diagnosis and treatment opportunity. A total of 67 specimens were embedded into cell blocks, and BAP1 immunocytochemistry (ICC) was performed. CDKN2A fluorescence in situ hybridization (FISH) was performed in 45 cases. The sensitivity, specificity and the association between the degree of cell atypia and the results of two auxiliary methods were analyzed. BAP1 ICC showed nonexpression in 13 of 24 cases of MM and 0 of 21 cases of benign mesothelial proliferation (BMP). The sensitivity was 54.2% (13/24), and the specificity was 100% (21/21). In addition, 22 metastatic adenocarcinoma (MA) cases all showed BAP1 expression. MM with BAP1 expression had more obvious cell atypia. CDKN2A deletion was found in 12 of 24 MM cases and 0 of 21 BMP cases. The sensitivity was 50% (12/24), and the specificity was 100% (21/21). BAP1 ICC and CDKN2A FISH are useful methods to differentiate MM from BMP. The cell atypia of MM with BAP1 expression was more obvious than MM with BAP1 nonexpression.

Autophagy-mediated compartmental cytoplasmic deletion is essential for tobacco pollen germination and male fertility.

In plants, macroautophagy/autophagy has mainly been associated with stress-related processes but how it impacts normal physiological and developmental processes remains largely unexplored. Pollen germination is the critical first step toward fertilization in flowering plants. It is metabolically demanding and relies on high levels of cytoplasmic reorganization activities to support a dramatic morphological transformation that underlies the development of a pollen tube as the conduit to deliver sperm for fertilization. The role of autophagy in this process remains unclear. Here we provide evidence that pollen germination is accompanied by elevated autophagic activity and successful pollen tube emergence depends on autophagy-mediated cytoplasmic deletion. Genetic and cytological experiments demonstrate that inhibition of autophagy prevents pollen germination while induces the persistence of a layer of undegraded cytoplasm at the germination aperture. Together, these results unveil a novel compartmentalized autophagy. Furthermore, high-throughput comparative lipidomic analyses show that suppressed autophagy-induced inhibition of pollen germination is accompanied by altered profiles of stored and signaling lipids. Proteomic analyses reveal that autophagy likely exert its role in pollen germination via downstream mitochondria-related pathways. These findings reveal a critical role for autophagy in initiating pollen germination and provide evidences for compartmental cytoplasmic deletion being crucial for male fertility. Abbreviations: 3-MA: 3-methyladenine; ATG: autophagy-related gene; Cer: ceramide; CL: cardiolipin; Con A: concanamycin A; DAG: diradylglycerol; GO: gene ontology; HAG: hour after germination; LC-MS: liquid chromatography-mass spectrometry; MAG: min after germination; MDC: monodansylcadaverine; PE: phosphatidylethanolamine; PI: phosphatidylinositol; PLD: phospholipase D; PtdIns3K: phosphatidylinositol 3-kinase; RT-qPCR: quantitative real-time reverse transcription PCR; TAG: triradylglycerol; TEM: transmission electron microscopy; TMT: tandem mass tagging.

Exercise Preconditioning Protects against Acute Cardiac Injury Induced by Lipopolysaccharide Through General Control Nonderepressible 2 Kinase.

Exercise preconditioning may protect against cardiac injury induced by lipopolysaccharide (LPS), but the mechanism is unresolved. The aim of this study is to explore whether the general control nonderepressible 2 (GCN2) kinase gene is associated with the protective effect of exercise preconditioning. Eight-week-old male C57BL/6J (n = 40) and GCN2 knockout (KO) (n = 40) mice were divided into four groups: control, LPS (L), exercise preconditioning (E), and exercise preconditioning LPS (EL). Mice in the exercise groups performed exercise for eight weeks. After exercise, all mice were given an equal volume of LPS or saline (10 µg/g). We measured the cardiac function using echocardiography and then collected heart tissue. Exercise preconditioning improved cardiac inflammation (interleukin-6, tumor necrosis factor α) and cardiac dysfunction (ejection fraction, fraction shortening) in C57 mice induced by LPS and also decreased the expression levels of GCN2, phosphorylation of eukaryotic translation initiation factor 2α (p-eIF2α), and activating transcription factor 4 (ATF4). Moreover, GCN2 KO decreased inflammation and cardiac dysfunction induced by LPS in sedentary mice. The inflammation and cardiac dysfunction in the GCN2 KO EL group were lower than in the C57 EL group, and the expression of GCN2, p-eIF2α, and ATF4 in the GCN2 KO EL group was lower than in the C57 EL group. Exercise preconditioning alleviated cardiac injury induced by LPS. GCN2 KO also improved cardiac injury. Exercise preconditioning promoted the effect of GCN2 KO in alleviating cardiac injury, and the GCN2 and eIF2α/ATF4 pathways play an important role in the process.

Effectiveness of intracavitary electrocardiogram-guided peripherally inserted central catheter tip placement in premature infants: a multicentre pre-post intervention study.

This pre-post intervention study was conducted in Neonatal Intensive Care Units in two Chinese hospitals. The objective was to evaluate the effectiveness and safety of intracavitary electrocardiogram (IC-ECG)-guided peripherally inserted central catheter (PICC) placement and tip positioning in premature infants. A total of 161 premature infants who required a PICC were enrolled and divided into two groups: pre-intervention group (n = 83) from October 2017 to July 2018 and post-intervention IC-ECG group (n = 78) from August 2018 to March 2019. Nurses were trained from May 2018 to July 2018. The reposition rate in the IC-ECG group and pre-interventions group was 3.85% and 19.28%, respectively (OR 5.970; 95% CI 1.666-21.395; p = 0.002). More infants achieved optimal tip position at the first attempt in the IC-ECG group than the pre-intervention group (93.59% vs 73.49%; OR 0.190; 95%CI 0.068-0.531; p = 0.001). The overall catheter-related complications in the pre-intervention group were 14.46% compared to 3.84% in the IC-ECG group (OR 2.962; 95%CI 1.013-8.661; p = 0.040). However, no significant differences were observed between the individual complication leakage, phlebitis and catheter-related blood stream infection.Conclusions: IC-ECG-guided peripherally inserted central catheter placement and tip positioning technology might decrease reposition rates, achieve more accurate tip positioning at the first attempt and might reduce catheter-related complications in premature infants. Further robust RCTs are needed to confirm the effectiveness of IC-ECG-guided PICC placement and tip positioning in neonates.What is Known:• Chest radiography is the gold standard for tip position confirmation of peripherally inserted central catheter placement.• Studies in adult patients have shown that electrocardiogram guidance in the placement of central venous catheters can be beneficial, while evidence in neonates is limited.What is New:• Intracavitary electrocardiogram-guided peripherally inserted central catheter placement might be superior to chest radiography in preterm infants.• Decreasing the repositioning rates and correct tip position of peripherally inserted central catheters might reduce catheter-related complications.