2X-Rhodamine: A Bright and Fluorogenic Scaffold for Developing Near-Infrared Chemigenetic Indicators.

Chemigenetic fusion of synthetic dyes with genetically encoded protein tags presents a promising avenue for in vivo imaging. However, its full potential has been hindered by the lack of bright and fluorogenic dyes operating in the "tissue transparency" near-infrared window (NIR, 700-1700 nm). Here, we report 2X-rhodamine (2XR), a novel bright scaffold that allows for the development of live-cell-compatible, NIR-excited variants with strong fluorogenicity beyond 1000 nm. 2XR utilizes a rigidified π-skeleton featuring dual atomic bridges and functions via a spiro-based fluorogenic mechanism. This design affords longer wavelengths, higher quantum yield (ΦF = 0.11), and enhanced fluorogenicity in water when compared to the phosphine oxide-cored, or sulfone-cored rhodamine, the NIR fluorogenic benchmarks currently used. We showcase their bright performance in video-rate dynamic imaging and targeted deep-tissue molecular imaging in vivo. Notably, we develop a 2XR variant, 2XR715-HTL, an NIR fluorogenic ligand for the HaloTag protein, enabling NIR genetically encoded calcium sensing and the first demonstration of in vivo chemigenetic labeling beyond 1000 nm. Our work expands the library of NIR fluorogenic tools, paving the way for in vivo imaging and sensing with the chemigenetic approach.

Brain magnetic resonance imaging as predictors in pediatric anti-N-methyl-D-aspartate receptor encephalitis.

OBJECTIVE: To investigate the associations between brain magnetic resonance imaging (MRI) changes and clinical profiles in children with anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. METHODS: Clinical data and brain MRI results of children diagnosed with anti-NMDAR encephalitis in Guangzhou Women and Children's Medical Center from October 2014 to June 2022 were retrospectively studied. RESULTS: A total of 143 children (Male: female 54:89) were enrolled, with a mean onset age of 6.8 years (6.8 ± 3.1). 40.6 % (58/143) of patients had abnormal initial brain MRI. Lesions in temporal lobe (34.5 %, 20/58) and frontal lobe (25.9 %, 15/58) were relatively common. Children with abnormal initial brain MRI were prone to have fever (P = 0.023), dystonia (P = 0.037), positive MOG antibodies (P = 0.015), higher cerebrospinal fluid (CSF) white blood cell count (WBC) (P = 0.019) and to receive rituximab treatment (P = 0.037). There were no significant differences in modified Rankin Scale (mRS) scores before immunotherapy, after immunotherapy and at last follow-up between the normal initial brain MRI group and abnormal group. No initial brain MRI changes were found to be associated with relapses. Brain MRI was reviewed in 72 patients at last follow-up with a median follow-up time of 25.5 months and 48.6 % (35/72) of patients had abnormal brain MRI. The mRS score of the group with normal brain MRI at last follow-up was significantly lower than that of the abnormal group. CONCLUSIONS: About 40.0 % of children with anti-NMDAR encephalitis had abnormal initial brain MRI. Initial brain MRI was associated with certain clinical profiles, but not with relapse and prognosis. Around half of patients had abnormal brain MRI at last follow-up and were prone to have higher mRS score.

Satralizumab as an add-on treatment in refractory pediatric AQP4-antibody-positive neuromyelitis optica spectrum disorder: a case report.

Neuromyelitis optica spectrum disorder (NMOSD) is a rare autoimmune disease of the central nervous system. Relapse and incomplete recovery from relapse are common in NMOSD. Most patients with NMOSD have IgG to aquaporin-4 (AQP4-IgG). New biological agents for AQP4-IgG-seropositive NMOSD, such as satralizumab, have become available for maintenance therapy. Satralizumab is an anti-interleukin-6 receptor monoclonal antibody. To date, few studies have evaluated satralizumab as an add-on treatment in pediatric NMOSD patients. Here, we report an 11-year-old girl with NMOSD who frequently relapsed under long-term treatment, including oral prednisone, rituximab, mycophenolate mofetil (MMF), and maintenance intravenous immunoglobulin treatment even with B-cell depletion. For the poor treatment response and to improve the efficacy of relapse prevention further, the patient received satralizumab treatment as an add-on therapy to MMF plus oral prednisone, with a dose of 120 mg administered subcutaneously at weeks 0, 2, and 4 and every 4 weeks after that. After initiating satralizumab, the patient remained relapse-free for 14 months at the last follow-up. Satralizumab might be effective and safe as an add-on treatment in refractory pediatric AQP4-IgG-seropositive NMOSD under B-cell depletion.

Epidemiology Trend of Urogenital System Disorders of Hospitalized Patients in an Eastern Chinese City from 2017 to 2019.

OBJECTIVE: To evaluate trends in the prevalence and clinical characteristics of urogenital diseases in hospitalized patients of secondary and tertiary hospitals in Ningbo, an east coast city in China, from 2017 to 2019. METHODS: We collected the data on hospitalized patients in Ningbo secondary and tertiary hospitals from January 1, 2017 to December 31, 2019. The data included age, sex, and diagnosis identified using the International Classification of Diseases (ICD) codes, which were obtained from Ningbo National Health Information Platform. We quantified the epidemiology (age/sex-specific) trend of urogenital system disorders. RESULTS: From January 2017 to December 2019, there were 256750 hospitalized patients with urogenital system disorders. These hospitalized patients comprised more women than men (1.45:1.00). The number of hospitalized patients with these diseases significantly increased over the 3 years (77505, 89167, and 90078, respectively; Z = 20.03, p < 0.001). The highest prevalence of these diseases was in the 40- to 64-year-old age group (47.37%), followed by the 18- to 39-year-old age group (23.94%). Over the 3 years, the five most common diseases in hospitalized male patients were male reproductive organ disorders, urolithiasis, tubulointerstitial disease, renal failure, and glomerular disease; Whereas the five most common diseases in hospitalized female patients were non-inflammatory disorders of the female genital tract, benign or dynamic undetermined tumors of the female reproductive organs, disorders of breast (according to ICD-10, disorders of breast (N60-N64) were involved in urogenital system diseases (N00-N99)), inflammatory diseases of female pelvic organs, and renal tubulointerstitial disease. In addition, the number of inpatients with renal tubulointerstitial disease significantly increased from 5952 to 9616 over the 3 years (rank increased from 6 to 3). CONCLUSIONS: Patients with urogenital system disorders significantly increased over the 3 years, occurring more often in women and most commonly in young and middle-aged people, which warrants more attention in clinical practice.

Deubiquitinase OTUD5 as a Novel Protector against 4-HNE-Triggered Ferroptosis in Myocardial Ischemia/Reperfusion Injury.

Despite the development of advanced technologies for interventional coronary reperfusion after myocardial infarction, a substantial number of patients experience high mortality due to myocardial ischemia-reperfusion (MI/R) injury. An in-depth understanding of the mechanisms underlying MI/R injury can provide crucial strategies for mitigating myocardial damage and improving patient survival. Here, it is discovered that the 4-hydroxy-2-nonenal (4-HNE) accumulates during MI/R, accompanied by high rates of myocardial ferroptosis. The loss-of-function of aldehyde dehydrogenase 2 (ALDH2), which dissipates 4-HNE, aggravates myocardial ferroptosis, whereas the activation of ALDH2 mitigates ferroptosis. Mechanistically, 4-HNE targets glutathione peroxidase 4 (GPX4) for K48-linked polyubiquitin-related degradation, which 4-HNE-GPX4 axis commits to myocyte ferroptosis and forms a positive feedback circuit. 4-HNE blocks the interaction between GPX4 and ovarian tumor (OTU) deubiquitinase 5 (OTUD5) by directly carbonylating their cysteine residues at C93 of GPX4 and C247 of OTUD5, identifying OTUD5 as the novel deubiquitinase for GPX4. Consequently, the elevation of OTUD5 deubiquitinates and stabilizes GPX4 to reverse 4-HNE-induced ferroptosis and alleviate MI/R injury. The data unravel the mechanism of 4-HNE in GPX4-dependent ferroptosis and identify OTUD5 as a novel therapeutic target for the treatment of MI/R injury.

Epidemiology Trend of Urogenital System Disorders of Hospitalized Patients in an Eastern Chinese City from 2017 to 2019

Objective: To evaluate trends in the prevalence and clinical characteristics of urogenital diseases in hospitalized patients of secondary and tertiary hospitals in Ningbo, an east coast city in China, from 2017 to 2019. Methods: We collected the data on hospitalized patients in Ningbo secondary and tertiary hospitals from January 1, 2017 to December 31, 2019. The data included age, sex, and diagnosis identified using the International Classification of Diseases (ICD) codes, which were obtained from Ningbo National Health Information Platform. We quantified the epidemiology (age/sex-specific) trend of urogenital system disorders. Results: From January 2017 to December 2019, there were 256750 hospitalized patients with urogenital system disorders. These hospitalized patients comprised more women than men (1.45:1.00). The number of hospitalized patients with these diseases significantly increased over the 3 years (77505, 89167, and 90078, respectively; Z = 20.03, p < 0.001). The highest prevalence of these diseases was in the 40- to 64-year-old age group (47.37%), followed by the 18- to 39-year-old age group (23.94%). Over the 3 years, the five most common diseases in hospitalized male patients were male reproductive organ disorders, urolithiasis, tubulointerstitial disease, renal failure, and glomerular disease; Whereas the five most common diseases in hospitalized female patients were non-inflammatory disorders of the female genital tract, benign or dynamic undetermined tumors of the female reproductive organs, disorders of breast (according to ICD-10, disorders of breast (N60–N64) were involved in urogenital system diseases (N00–N99)), inflammatory diseases of female pelvic organs, and renal tubulointerstitial disease. In addition, the number of inpatients with renal tubulointerstitial disease significantly increased from 5952 to 9616 over the 3 years (rank increased from 6 to 3) (AU)

Numerical Simulation of Temperature Field Optimization to Enhance Nitrogen Transfer in GaN Crystal Growth by the Na-Flux Method.

Improved nitrogen transport is crucial for enhancing the growth rate of GaN crystals using the Na-flux method. This study investigates the nitrogen transport mechanism during the growth of GaN crystals by the Na-flux method using a combination of numerical simulations and experiments. The results indicate that the temperature field affects the effect of nitrogen transfer, and we propose a novel bottom ring heating approach to optimize the temperature field and enhance nitrogen transfer during the growth of GaN crystals. The simulation results demonstrate that optimizing the temperature field improves nitrogen transfer by causing convection within the melt to float up from the crucible wall and sink at the crucible center. This enhancement improves the nitrogen transfer from the gas-liquid interface to the GaN crystal growth surface, thereby accelerating the growth rate of GaN crystals. Additionally, the simulation results indicate that the optimized temperature field substantially reduces polycrystalline generation at the crucible wall. These findings are also a realistic guide to the growth of other crystals in the liquid phase method.

Pediatric myelin oligodendrocyte glycoprotein antibody-associated disease in southern China: analysis of 93 cases.

Objective: To study the clinical features of children diagnosed with myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) in southern China. Methods: Clinical data of children diagnosed with MOGAD from April 2014 to September 2021 were analyzed. Results: A total of 93 children (M/F=45/48; median onset age=6.0 y) with MOGAD were involved. Seizures or limb paralysis was the most common onset or course symptom, respectively. The most common lesion locations in brain MRI, orbital MRI, and spinal cord MRI were basal ganglia and subcortical white matter, the orbital segment of the optic nerve, and the cervical segment, respectively. ADEM (58.10%) was the most common clinical phenotype. The relapse rate was 24.7%. Compared with the patients without relapse, relapsed patients had a longer interval from onset to diagnosis (median: 19 days VS 20 days) and higher MOG antibody titer at onset (median: 1:32 VS 1:100) with longer positively persistent (median: 3 months VS 24 months). All patients received IVMP plus IVIG at the acute phase, and 96.8% of patients achieved remission after one to three courses of treatment. MMF, monthly IVIG, and maintaining a low dose of oral prednisone were used alone or in combination as maintenance immunotherapy for relapsed patients and effectively reduced relapse. It transpired 41.9% of patients had neurological sequelae, with movement disorder being the most common. Compared with patients without sequelae, patients with sequelae had higher MOG antibody titer at onset (median: 1:32 VS 1:100) with longer persistence (median: 3 months VS 6 months) and higher disease relapse rate (14.8% VS 38.5%). Conclusions: Results showed the following about pediatric MOGAD in southern China: the median onset age was 6.0 years, with no obvious sex distribution difference; seizure or limb paralysis, respectively, are the most common onset or course symptom; the lesions of basal ganglia, subcortical white matter, the orbital segment of the optic nerve, and cervical segment were commonly involved in the CNS MRI; ADEM was the most common clinical phenotype; most had a good response to immunotherapy; although the relapse rate was relatively high, MMF, monthly IVIG and a low dose of oral prednisone might effectively reduce relapse; neurological sequelae were common, and possibly associated with MOG antibody status and disease relapse.

Prognostic impact of tumor deposits on overall survival in colorectal cancer: Based on Surveillance, Epidemiology, and End Results database.

BACKGROUND: In colorectal cancer, tumor deposits (TDs) are considered to be a prognostic factor in the current staging system, and are only considered in the absence of lymph node metastases (LNMs). However, this definition and the subsequent prognostic value based on it is controversial, with various hypotheses. TDs may play an independent role when it comes to survival and addition of TDs to LNM count may predict the prognosis of patients more accurately. AIM: To assess the prognostic impact of TDs and evaluate the effect of their addition to the LNM count. METHODS: The patients are derived from the Surveillance, Epidemiology, and End Results database. A prognostic analysis regarding impact of TDs on overall survival (OS) was performed using Cox regression model, and other covariates associating with OS were adjusted. The effect of addition of TDs to LNM count on N restaging was also evaluated. The subgroup analysis was performed to explore the different profile of risk factors between patients with and without TDs. RESULTS: Overall, 103755 patients were enrolled with 14131 (13.6%) TD-positive and 89624 (86.4%) TD-negative tumors. TD-positive patients had worse prognosis compared with TD-negative patients, with 3-year OS rates of 47.3% (95%CI, 46.5%-48.1%) and 77.5% (95%CI, 77.2%-77.8%, P < 0.0001), respectively. On multivariable analysis, TDs were associated poorer OS (hazard ratio, 1.35; 95%CI, 1.31-1.38; P < 0.0001). Among TD-positive patients, the number of TDs had a linear negative effect on disease-free survival and OS. After reclassifying patients by adding TDs to the LNM count, 885 of 19 965 (4.4%) N1 patients were restaged as pN2, with worse outcomes than patients restaged as pN1 (3-year OS rate: 78.5%, 95%CI, 77.9%-79.1% vs 63.2%, 95%CI, 60.1%-66.5%, respectively; P < 0.0001). CONCLUSION: TDs are an independent prognostic factor for OS in colorectal cancer. The addition of TDs to LNM count improved the prognostic accuracy of tumor, node and metastasis staging.

Stable Monodisperse Pb1-x Cdx S Quantum Dots for NIR-II Bioimaging by Aqueous Coprecipitation of Bimetallic Clusters.

Aqueous lead sulfide (PbS) quantum dots (QDs) synthesized by traditional methods are unstable, so that they are usually coated with cadmium sulfide (CdS) to prevent oxidation, which are complicated and not satisfactory for mass production. Here, stable ternary Pb1-x Cdx S QDs were synthesized by in situ coprecipitation of Pb4-n Cdn O4 bimetallic clusters in an aqueous solution, which possess a uniform size of 4.0±0.2 nm and the second near-infrared (NIR-II) emission at 1100 nm with photoluminescence quantum yield (PLQY) as high as 17.72 %. Stored at 4 °C and in colloidal form, the PLQY of Pb1-x Cdx S QDs remained at 90.9 % of the initial value after 15 days, while stored as powder, the spectra did not change after 5 months. The high PLQY and good water compatibility of Pb1-x Cdx S QDs provide a good performance in vivo vasculature imaging and lymphatic system imaging at a very low power density (10 mW cm-2 ) in the NIR-II window.

Ganoderic acid A improves osteoarthritis by regulating RANKL/OPG ratio.

Ganoderma mushrooms have been used to treat rheumatoid arthritis (RA) in East Asia. Whether Ganoderic acid A (GAA), the natural product extracted from Ganoderma, could be utilized to alleviate osteoarthritis (OA) is investigated in this study. Destabilization of the medial meniscus (DMM) model was constructed to reveal the in vivo effect of GAA. We found that GAA could significantly alleviate the pathology of DMM, as confirmed by the diminished maximum histologic scores. On the contrary, GAA could down-regulate the relative expression of osteoprotegerin (OPG) and up-regulate the relative expression of nuclear factor kappa-B ligand (RANKL) in DMM cartilage and human articular chondrocytes (HC-A) cells with diminished matrix metallopeptidase 13 (MMP-13) secretion in the synovial fluid. It was further demonstrated that the serum concentration of OPG was correlated with the severity of osteoarthritis. All these data reveal that GAA could improve OA by regulating the RANKL/OPG ratio to inhibit the secretion of MMP-13.

Reduced Lon protease 1 expression in podocytes contributes to the pathogenesis of podocytopathy.

Emerging evidence has shown that mitochondrial dysfunction is closely related to the pathogenesis of podocytopathy, but the molecular mechanisms mediating mitochondrial dysfunction in podocytes remain unclear. Lon protease 1 is an important soluble protease localized in the mitochondrial matrix, although its exact role in podocyte injury has yet to be determined. Here we investigated the specific role of this protease in podocyte in glomerular injury and the progression of podocytopathy using podocyte-specific Lon protease 1 knockout mice, murine podocytes in culture and kidney biopsy samples from patients with focal segmental glomerular sclerosis and minimal change disease. Downregulated expression of Lon protease 1 was observed in glomeruli of kidney biopsy samples demonstrating a negative correlation with urinary protein levels and glomerular pathology of patients with focal segmental glomerular sclerosis and minimal change disease. Podocyte-specific deletion of Lon protease 1 caused severe proteinuria, impaired kidney function, severe kidney injury and even mortality in mice. Mechanistically, we found that continuous podocyte Lon protease 1 ablation induced mitochondrial homeostasis imbalance and dysfunction, which then led to podocyte injury and glomerular sclerosis. In vitro experiments implicated the kidney protective effect of Lon protease 1, which inhibited mitochondrial dysfunction and podocyte apoptosis. Thus, our findings suggest that the regulation of Lon protease 1 in podocytes may provide a novel therapeutic approach for the podocytopathy.

Novel compound heterozygous TARS2 variants in a Chinese family with mitochondrial encephalomyopathy: a case report.

BACKGROUND: Mitochondrial encephalomyopathy caused by bi-allelic deleterious variants in TARS2 is rare. To date, only two pedigrees were reported in the literature and the connection between the gene and disease needs further study. CASE PRESENTATION: We report one infant who presented with limb hypertonia, epilepsy, developmental delay, and increased serum lactate from a non-consanguineous Chinese family. Whole-genome sequencing was performed to help to underlie the cause. We identified compound heterozygous variants c.470C > G, p.Thr157Arg and c.2143G > A, p.Glu715Lys in TARS2 and the variants were confirmed by Sanger sequencing. The patient was diagnosed with combined oxidative phosphorylation deficiency 21 according to the Online Mendelian Inheritance in Man (OMIM) database based on the clinical data and the deleterious effect of the two variants in TARS2 predicted by in silico tools. CONCLUSIONS: We presented one case diagnosed with combined oxidative phosphorylation deficiency 21 based on clinical characteristics and genetic analysis. This is the first case in China and the fourth case in the world based on our document retrieval. This study facilitates the understanding of combined oxidative phosphorylation deficiency disease and demonstrates that the next-generation sequencing has a high potential to study inherited disease with high phenotypic heterogeneity and genetic heterogeneity including mitochondrial diseases such as combined oxidative phosphorylation deficiency.

The Possible Role of Hypothalamus-Pituitary-Adrenal Dysfunction in Epileptic Spasms.

PURPOSE: To analyze diurnal cortisol (COR) rhythms among children with epileptic spasms (ESs) and explore the relationship between endocrine factors, circadian rhythm, and ES. METHODS: This study assessed the COR and adrenocorticotropic hormone (ACTH) levels at 08:00 and 16:00, and COR values at 00:00 among children with ESs. Additionally, the etiology of ESs was analyzed. All cases were divided into the following three etiology groups: genetic group, structural etiology group, and unknown etiology group. ACTH was administered to 24 patients, who were divided into the positive electroclinical outcome group and negative electroclinical outcome group. All data were analyzed using a two-way repeated measures analysis of variance. RESULTS: All children showed a COR rhythm. Controls displayed a significantly different COR rhythm from that in the ES group (Fgroup*COR =24.100, p = 0.000). It was observed that the ACTH levels at 08:00 (t = -3.720) and 16:00 (t=-3.794) and COR levels at 16:00 (t = -2.264) and 00:00 (t = -4.607) in the ES group were significantly higher than those in the control group (p < 0.05); COR levels at 08:00 were significantly lower among individuals in the structural etiology group (F = 3.828, p < 0.05). COR levels at 08:00 in the negative electroclinical outcome group (668.30 ± 227.42) nmol/L were higher than those in the positive electroclinical outcome group (462.25 ± 249.71) nmol/L. CONCLUSION: Our results suggest that the change in COR rhythm is an important pathophysiological characteristic of ESs, suggesting that hypothalamus-pituitary-adrenal axis dysfunction possibly leads to the different manifestations of ESs.

An enzyme-free amplification strategy based on two-photon fluorescent carbon dots for monitoring miR-9 in live neurons and brain tissues of Alzheimer's disease mice.

An enzyme-free amplification strategy based on two-photon fluorescent carbon dots for monitoring miR-9 in live neurons and brain tissues of Alzheimer's disease (AD) mice. Notably, using our developed probe, miR-9 was found to be up-regulated in early onset AD, while it was found to be down regulated to lower than the normal level in late onset AD.

EasyMEG: An easy-to-use toolbox for MEG analysis.

BACKGROUND AND OBJECTIVE: Magnetoencephalography (MEG) is an advanced magnetic source imaging technology that measures the magnetic fields produced by neural activities. It has been extensively used in scientific research and clinical diagnosis due to its high temporal and spatial resolution. Considering the special nature of MEG data, it needs to perform a series of processes and analysis to obtain valuable information. Therefore, the identification of data processing is a key point of MEG studies. At present, the software for MEG analysis such as FieldTrip has no Graphic User Interface (GUI) and users must write their own script to perform concrete analysis. It brings the difficulties to researchers like the doctors without experience in programming or newcomers to MEG. Thus, an open-sourced software-EasyMEG was developed. It has friendly interface with highly functions-integration. METHODS: The functions of EasyMEG are developed based on MATLAB language to ensure the consistency of the user interface under different operating systems. EasyMEG is a highly integrated software that contains a set of functions for preprocessing, time-lock analysis, time-frequency analysis, source analysis, and plotting. EasyMEG provides a friendly GUI and allows users to complete analyses through a simple and clean interface. RESULTS: This toolbox has been released as an open-source software on GitHub under the GNU General Public License: https://tonywu2018.github.io/EasyMEG/. CONCLUSIONS: We hope to improve this toolbox by the power of community and wish to make EasyMEG a simple and powerful toolbox for further MEG studies.

Xian-Ling-Gu-Bao induced inflammatory stress rat liver injury: Inflammatory and oxidative stress playing important roles.

ETHNOPHARMACOLOGICAL RELEVANCE: Xian-Ling-Gu-Bao (XLGB) Fufang is an herbal formula that has been used in clinical settings to treat osteoporosis, osteoarthritis, aseptic bone necrosis, and climacteric syndrome. Despite its uses, XLGB treatment has been linked to potential liver injury. To date, there is a lack of clear demonstration of such toxicity in animal models. AIM OF THE STUDY: As animal models fail to reproduce the XLGB hepatotoxicity reported in humans, because human hepatocytes are clearly more sensitive to XLGB, this study was designed to investigate a more reliable animal model of such toxicity. MATERIALS AND METHODS: We randomized rats into five groups, as follows: CON (control), XLGB, lipopolysaccharide (LPS), L-XLGB/LPS (XLGB, 0.125 g/kg; LPS, 0.1 mg/kg), and XLGB/LPS (XLGB, 1.25 g/kg; LPS, 0.1 mg/kg). These groups were treated with 0.5% sodium carboxymethyl cellulose (CMC-Na), XLGB suspension, normal saline, or LPS. The first administration of XLGB [0.125 g/kg or 1.25 g/kg, by mouth (PO)] or its solvent (0.5% CMC-Na) was delivered, and then food was removed. Twelve hours after the first administration of XLGB, rats received LPS [0.1 mg/kg, intravenously (IV)] or saline control. After 30 min, a second administration of XLGB (0.125 g/kg or 1.25 g/kg, PO) or solvent was administered. The rats were anesthetized at 12 h or 24 h following the second administration of XLGB. Liver function was evaluated by measuring liver weight, liver microscopy, serum biochemistry and plasma microRNA-122 (miR-122). The plasma levels of 27 cytokines were measured to evaluate inflammation. Moreover, the expression of cytochrome P450 2E1 (CYP2E1), nicotinic adenine dinucleotide phosphate (NADPH) oxidase and inducible nitric oxide synthase (iNOS) at protein levels were observed; immunofluorescence and immunohistochemistry were used to confirmed the hepatotoxicity of XLGB. RESULTS: Hepatotoxicity in male rats with moderate inflammation induced by XLGB was indicated by liver histopathology, serum biochemical analysis, serum miR-122 levels, and immunofluorescent assessments. We observed significant increases in liver weight and liver indexes in male rats with moderate inflammation in response to XLGB. Histopathological assessment further showed that extensive hepatocellular necrosis and inflammatory infiltration were evident in rats co-treated with XLGB/LPS. The levels of serum transaminases [alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT)], total bilirubin (TBIL) and triglyceride (TG), which are markers of liver function, were also significantly increased by XLGB/LPS treatment. Similarly, miR-122 was significantly elevated in XLGB/LPS treated rats relative to other groups. An immunofluorescent assessment showed extensive apoptosis in hepatocytes from these co-treated rats. What is more, XLGB can dose-dependently induce liver injury in male rats with moderate inflammation. Hepatic CYP2E1, neutrophil chemotactic factor (NCF-1), iNOS, and NOX-2 (an NADPH oxidase subunit) levels were increased in response to XLGB treatment, and staining for DMPO nitrone adducts further showed elevated oxidative stress level in XLGB/LPS-treated rats relative to the other experimental groups. CONCLUSION: LPS and XLGB co-treatment in rats led to marked hepatotoxicity. This toxicity was associated with disrupted lipid metabolism, extensive liver necrosis and inflammatory infiltration, apoptosis, and expression of oxidative stress-related proteins. These results demonstrate a valuable model for the study of iDILI in the context of XLGB treatment, and further provide insights into the potential mechanisms by which XLGB may induce hepatotoxicity in humans.

cybLuc: An Effective Aminoluciferin Derivative for Deep Bioluminescence Imaging.

To enhance the efficiency of firefly luciferase/luciferin bioluminescence imaging, a series of N-cycloalkylaminoluciferins (cyaLucs) were developed by introducing lipophilic N-cycloalkylated substitutions. The experimental results demonstrate that these cyaLucs are effective substrates for native firefly luciferase (Fluc) and can produce elevated bioluminescent signals in vitro, in cellulo, and in vivo. It should be noted that, in animal studies, N-cyclobutylaminoluciferin (cybLuc) at 10 µM (0.1 mL), which is 0.01% of the standard dose of d-luciferin (dLuc) used in mouse imaging, can radiate 20-fold more bioluminescent light than d-luciferin (dLuc) or aminoluciferin (aLuc) at the same concentration. Longer in vivo emission imaging using cybLuc suggests that it can be used for long-time observation. Regarding the mechanism of cybLuc, our cocrystal structure data from firefly luciferase with oxidized cybLuc suggested that oxidized cybLuc fits into the same pocket as oxyluciferin. Most interestingly, our results demonstrate that the sensitivity of cybLuc in brain tumor imaging contributes to its extended application in deep tissues.

Bioluminogenic Imaging of AminopeptidaseN In Vitro and In Vivo.

Bioluminescence is a process that converts biochemical energy to visible light. Bioluminescence-based imaging technology has been widely applicable in the imaging of process envisioned in life sciences. As one of the most popular bioluminescence system, the firefly luciferin-luciferase system is exceptionally suitable for deep tissue imaging in living animals owing to its long wavelength emission light. Herein, we report the experimental detail of bioluminogenic imaging of aminopeptidase N activity both in vitro and in vivo.