Objective Intracranial hemorrhage(ICH),the second most common subtype of stroke,exacerbates the disruption of the blood-brain barrier(BBB),leading to vasogenic edema,plasma protein extravasation,and infiltration of neurotoxic substances.The clearance capacity of the brain plays a crucial role in maintaining BBB homeostasis and facilitating patient recovery after hemorrhage.This study aimed to investigate the effect of circadian rhythms on BBB function,neuronal damage,and clearance capabilities. Methods The transwell model and hemoglobin were co-cultured to simulate the BBB environment after ICH.After intervention with different light groups,neuronal apoptosis was determined,glial phagocytosis was analyzed,the expression of endogenous clearing-related proteins aquaporin 4(AQP4)and low-density lipoprotein receptor-related protein 1(LRP1)was detected by western blotting and immunofluorescence dual standard method,and the expression of the tight junction protein occludin and melatonin receptor 1A(MTNR1A)was quantitatively analyzed. Results Circadian rhythms play a key role in maintaining the integrity of the BBB,reducing oxidative stress-induced neuronal damage,and improving microglial phagocytosis.Meanwhile,the expression of occludin and MTNR1A in neurovascular unit(NVU)co-cultured with hemoglobin improved the expression of AQP4 and LRP1,the key proteins in the NVU's endogenous brain clearance system. Conclusion Circadian rhythm(alternating black and white light)protects the NVU BBB function after ICH,promotes the expression of proteins related to the clearance of the hematoma,provides new evidence for the clinical treatment of patients recovering from ICH,and improves the circadian rhythm to promote brain metabolism and hematoma clearance.

Humans ; Transcriptome/genetics* ; East Asian People ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Pulmonary Disease, Chronic Obstructive/genetics* ; Polymorphism, Single Nucleotide/genetics*

BACKGROUND: Pulmonary microvascular endothelial cells (PMVECs) were not complex, and the endothelial barrier was destroyed in the pathogenesis progress of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). Previous studies have demonstrated that hepatocyte growth factor (HGF), which was secreted by bone marrow mesenchymal stem cells, could decrease endothelial apoptosis. We investigated whether mTOR/STAT3 signaling acted in HGF protective effects against oxidative stress and mitochondria-dependent apoptosis in lipopolysaccharide (LPS)-induced endothelial barrier dysfunction and ALI mice. METHODS: In our current study, we introduced LPS-induced PMEVCs with HGF treatment. To investigate the effects of mammalian target of rapamycin (mTOR)/signal transducer and activator of transcription 3 (STAT3) pathway in endothelial oxidative stress and mitochondria-dependent apoptosis, mTOR inhibitor rapamycin and STAT3 inhibitor S3I-201 were, respectively, used to inhibit mTOR/STAT3 signaling. Moreover, lentivirus vector-mediated mTORC1 (Raptor) and mTORC2 (Rictor) gene knockdown modifications were introduced to evaluate mTORC1 and mTORC1 pathways. Calcium measurement, reactive oxygen species (ROS) production, mitochondrial membrane potential and protein, cell proliferation, apoptosis, and endothelial junction protein were detected to evaluate HGF effects. Moreover, we used the ALI mouse model to observe the mitochondria pathological changes with an electron microscope in vivo. RESULTS: Our study demonstrated that HGF protected the endothelium via the suppression of ROS production and intracellular calcium uptake, which lead to increased mitochondrial membrane potential (JC-1 and mitochondria tracker green detection) and specific proteins (complex I), raised anti-apoptosis Messenger Ribonucleic Acid level (B-cell lymphoma 2 and Bcl-xL), and increased endothelial junction proteins (VE-cadherin and occludin). Reversely, mTOR inhibitor rapamycin and STAT3 inhibitor S3I-201 could raise oxidative stress and mitochondria-dependent apoptosis even with HGF treatment in LPS-induced endothelial cells. Similarly, mTORC1 as well as mTORC2 have the same protective effects in mitochondria damage and apoptosis. In in vivo experiments of ALI mouse, HGF also increased mitochondria structural integrity via the mTOR/STAT3 pathway. CONCLUSION In all, these reveal that mTOR/STAT3 signaling mediates the HGF suppression effects to oxidative level, mitochondria-dependent apoptosis, and endothelial junction protein in ARDS, contributing to the pulmonary endothelial survival and barrier integrity.
Animals ; Apoptosis ; Calcium/metabolism* ; Endothelial Cells/metabolism* ; Endothelium/metabolism* ; Hepatocyte Growth Factor/metabolism* ; Lipopolysaccharides/pharmacology* ; Mammals/metabolism* ; Mechanistic Target of Rapamycin Complex 1/metabolism* ; Mechanistic Target of Rapamycin Complex 2/metabolism* ; Mice ; Mitochondria/metabolism* ; Oxidative Stress ; Reactive Oxygen Species/metabolism* ; Respiratory Distress Syndrome ; Sirolimus/pharmacology* ; TOR Serine-Threonine Kinases/metabolism*

AIM: To establish an LC-MS/MS method to determine the concentration of ambrisentan in human plasma and apply it to the study of human pharmacokinetics. METHODS: After extracting ambrisentan and internal standard from human plasma by liquid-liquid extraction, chromatographic separation was performed on a Waters Symmetry C

Objective:To investigate the characteristics of polypoidal lesions and abnormal branching vascular network (BVN) in patients with polypoidal choroidal vasculopathy (PCV) by optical coherence tomography angiography (OCTA).Methods:A case series study was conducted.Untreated 45 eyes of 42 patients with initial diagnosis of PCV from May 2016 to April 2017 in Peking Union Medical College Hospital were enrolled.There were 26 males (27 eyes) and 16 females (18 eyes) with the average age of (68.59±8.52) years.All patients underwent fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA) and OCTA.The OCTA image features and detection rate of polypoidal lesion and BVN were analyzed.The study protocol was approved by the Ethics Committee of Peking Union Medical College Hospital (No.S-K631). Written informed consent was obtained from each patient prior to any examination.Results:ICGA images showed obvious polyps with higper fluorescence in all PCV patients, and BVN structure was observed in the ICGA images of 38 eyes.Ring, circle, cluster or node like hyper-reflective structures were detected in polypoidal lesions in the en face OCTA images of 35 eyes, and the detection rate was 77.8% (35/45). The BVN structure was detected in the OCTA images of all the 38 eyes, with the 100% (38/38) detection rate.Conclusion:OCTA can detect polypoidal lesions and BVN of PCV non-invasively, which can locate the lesion in combination with en face image and B-scan image.

Objective: To evaluate the sensitivity and specificity of spectral domain optical coherence tomography (SD-OCT) for the differentiation of PCV from wet age-related macular degeneration (wAMD). Methods: A cross-sectional study was conducted from Jun 2014 to Apr 2016 at Peking Union Medical College Hospital.The case series included 132 eyes of 126 patients with clinical diagnosis of PCV or wAMD.Eyes with three or more of the following SD-OCT findings were diagnosed with PCV: double layer sign, pigment epithelium detachment (PED), a sharp PED peak, a PED notch and a hyporeflective lumen representing polypoidal lesion.The sensitivity and specificity of the OCT-based diagnosis was estimated.Levels of agreement were determined by κ analyses. Results: One hundred and twenty-six patients (132 eyes) with PCV or wAMD were enrolled in the study.In PCV patients, PED notch, a hyporeflective lumen representing polypoidal lesion, PED peak, double layer sign and PED were 38, 35, 48, 40 and 46, accounting for 67.9%, 62.5%, 85.7%, 71.4% and 82.1%, respectively.In wAMD patients, PED notch, a hyporeflective lumen representing polypoidal lesion, PED peak, double layer sign and PED were 12, 3, 11, 8 and 26, accounting for 14.5%, 10.5%, 34.2%, 15.8% and 3.9%, respectively, which were all lower than PCV, with statistically significant differences (all at P<0.001). SD-OCT detected PCV in 49 of 56 eyes and 10 PCV patients were misdiagnosed as wAMD.The sensitivity and specificity of SD-OCT for the differentiation of PCV from wAMD were 87.5% and 86.8%, respectively.The consistence between SD-OCT and imdocyanine green angiography (ICGA) was moderate (κ=0.738, P<0.001). Conclusions SD-OCT exhibits high sensitivity and specificity in PCV diagnosis.The presence of DLS, PED, a sharp PED peak, a PED notch, and a hyporeflective lumen representing polypoidal lesion is suggested to be a new diagnostic strategy for PCV.

Objective:To explore the pathogenic variants of a family with syndromic deafness by high-throughput sequencing.Methods:The family was from Puyang City, Henan Province, and had four members, including two with syndromic deafness. The proband and his sister had congenital deafness, and their parents had normal phenotypes. The clinical phenotype of the family was characterized using clinical examinations and pedigree analysis. The clinical examinations included imaging examination, audiometry (pure tone audiometry, acoustic immittance, brainstem auditory evoked potential, and otoacoustic emission), vestibular function test, and ophthalmic examination (visual acuity test, visual field test, fundus examination, visual evoked potential, and electroretinogram). Target exome sequencing of 129 known deafness genes and bioinformatics analysis were used to screen suspected pathogenic variants. Sanger sequencing and minigene assay were used to verify and functionally investigate the mutation detected, respectively. According to the standards and guidelines for interpreting genetic variants proposed by the American College of Medical Genetics and Genomics, the variants c.6049G>A and c.8699A>G were classified as pathogenic/likely pathogenic, and the variant c.9856C>G was classified as variants of uncertain significance.Results:The probands and his sister had severe sensorineural hearing loss with decreased binocular vision, night blindness, decreased peripheral visual field sensitivity and partial visual field defect, and normal vestibular function. Both of them had three CDH23 mutations, including CDH23 (NM_022124.5) c.6049G>A (p.Gly2017Ser),c.9856C>G (p.His3286Asp), and c.8699A>G (p. Asp2900Gly), The first two were inherited from the father, and the last one was from the mother. The missense variants c.9856C>G and c.8699A>G were not included in the gnomad database. The missense mutation c.6049G>A was located in the last position of exon 46 and was predicted to affect splicing by bioinformatics software. The minigene experiment showed that the mutation cause exon skipping of exon 46, resulting in an abnormal protein. Conclusions:Compound heterozygous variations of the CDH23 are the leading cause of USH1D in the family. This study confirms that the compound heterozygosity of splicing and missense variants of the CDH23 gene could lead to USH1D.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Objective To use the "three-dimensional quality structure" model as a theoretical basis, based on multiple evidence-based evidence, to construct ICU nursing sensitivity quality indicators, and to provide reference for the evaluation and monitoring of ICU department nursing safety quality. Methods Evidence-based and Delphi methods were used to complete 3 rounds of inquiry among 27 experts to obtain consensus opinions of experts and to determine nursing quality indicators. Results Of the 30 articles included, 17 wereⅢA, 10 wereⅣA, 2 wereⅤA, and 1 wasⅤB. According to the"three-dimensional structure theory"model, the included indicators were sorted out and a total of 12 items were obtained. The effective recovery rate of 3 rounds of questionnaires was 96.6％, 100.0％, and 96.4％. The authoritative coefficients were 0.899, 0.895, and 0.909, and the coordination coefficients of the first, second, and third indexes were 0.340, 0.379, and 0.403, respectively,with significance (P<0.01). The final ICU nursing sensitivity quality indicator consists of 3 first-level indicators, 15 second-level indicators and 69 third-level indicators. The selection of indicators at all levels gradually converges. Conclusions The established ICU nursing sensitivity quality index accords with the principle of scientificity and practicality, and can be used for ICU department nursing quality assessment.

Vasovagal syncope(VVS),with its relatively high morbidity,is one of the most common types of au-tonomic-mediated reflex syncope in children,which can seriously affect pediatric patients in their life and study.A se-ries of research have shown that the mechanism for VVS is closely related to autonomic nervous imbalance,neurohor-monal factor,and cerebral blood flow abnormality,and has a certain degree of genetic tendency.Now the progress in the mechanisms for VVS at home and abroad is reviewed,in order to provide certain help for further research.