Successful Treatment of Rituximab-Induced Crohn's Disease With Ustekinumab.

Autoimmune optic papillitis is a rare disorder that causes progressive visual loss, often treated with rituximab (RTX). However, its use is not without risks. Here, we present a 51-year-old man who experienced vision loss because of autoimmune optic papillitis, which was well-controlled with RTX. Four years later, the patient developed abdominal pain and diarrhea and was found to have RTX-induced Crohn's disease (CD). The patient failed treatment with azathioprine, but was subsequently able to achieve clinical and endoscopic remission of his CD with ustekinumab, while continuing RTX therapy for autoimmune optic papillitis. This case report describes the efficacy of the anti-interleukin 12/23 monoclonal antibody in inducing remission of RTX-induced CD.

Lymphocytic Colitis With Enterocolic Lymphocytic Phlebitis.

Enterocolic lymphocytic phlebitis is a rare lymphocytic vasculitis afflicting the gastrointestinal veins without involving the arterial system. Lymphocytic colitis is a more common pathology described as lymphocytic inflammation of the colonic epithelium. Concurrence of both these pathologies is extremely rare. We describe a 53-year-old man presenting with chronic watery diarrhea, abdominal pain, and weight loss. Colonoscopic examination revealed normal-appearing mucosa, but biopsy findings revealed lymphocytic colitis with coexisting enterocolic lymphocytic phlebitis. The patient was started on oral budesonide and responded to the treatment with symptomatic and histopathological resolution.

Enhancing bacterial cellulose production with hypoxia-inducible factors.

Komagataeibacter xylinus is an aerobic strain that produces bacterial cellulose (BC). Oxygen levels play a critical role in regulating BC synthesis in K. xylinus, and an increase in oxygen tension generally means a decrease in BC production. Fumarate nitrate reduction protein (FNR) and aerobic respiration control protein A (ArcA) are hypoxia-inducible factors, which can signal whether oxygen is present in the environment. In this study, FNR and ArcA were used to enhance the efficiency of oxygen signaling in K. xylinus, and globally regulate the transcription of the genome to cope with hypoxic conditions, with the goal of improving growth and BC production. FNR and ArcA were individually overexpressed in K. xylinus, and the engineered strains were cultivated under different oxygen tensions to explore how their overexpression affects cellular metabolism and regulation. Although FNR overexpression did not improve BC production, ArcA overexpression increased BC production by 24.0% and 37.5% as compared to the control under oxygen tensions of 15% and 40%, respectively. Transcriptome analysis showed that FNR and ArcA overexpression changed the way K. xylinus coped with oxygen tension changes, and that both FNR and ArcA overexpression enhanced the BC synthesis pathway. The results of this study provide a new perspective on the effect of oxygen signaling on growth and BC production in K. xylinus and suggest a promising strategy for enhancing BC production through metabolic engineering. KEY POINTS: • K. xylinus BC production increased after overexpression of ArcA • The young's modulus is enhanced by the ArcA overexpression • ArcA and FNR overexpression changed how cells coped with changes in oxygen tension.

Macular hole following phakic intraocular lens implantation: A case report.

BACKGROUND: Phakic intraocular lens (pIOL) implantation has been commonly prescribed and is considered as a safe and effective option for correcting high myopia. However, it is associated with multiple complications. CASE SUMMARY: This report describes a case of full-thickness macular hole (MH) in a patient with a history of bilateral pIOL implantation for the correction of myopia of -12.00 diopters in both eyes 7 mo ago. The MH closed after pars plana vitrectomy with internal limiting membrane removal and the best-corrected visual acuity improved to 20/40 in the left eye. CONCLUSION: In rare cases, MH can occur following pIOL. In this present case report, we analyzed the formation process of MH following the surgery and emphasized that it is important to inform highly myopic patients about the risk of MH occurrence while being aware of the symptoms of this complication.

Fabrication of Protonated Two-Dimensional Metal-Organic Framework Nanosheets for Highly Efficient Iodine Capture from Water.

Radioactive iodine (129I and 131I), produced or released from nuclear-related activities, posed severe effects on both human health and environment. The efficient removal of radioiodine from aqueous medium and vapor phase is of paramount importance for the sustainable development of nuclear energy. Herein, a metal-organic framework (MOF) nanosheet with a positive charge was constructed for the capture of iodine for the first time. The as-synthesized ultrathin nanosheets, with a thickness of 4.4 ± 0.1 nm, showed a record-high iodine adsorption capacity (3704.08 mg g-1) from aqueous solution, which is even higher than that from the vapor phase (3510.05 mg g-1). It can be ascribed to the fully interactions between the extensive accessible active sites on the largely exposed surface of 2D MOF nanosheets and the target pollutants, which also gave rise to fast adsorption kinetics with relative high removal efficiencies in the low concentrations, even in seawater. Moreover, a facile recyclability with fast desorption kinetics can also be achieved for the MOF nanosheets. The excellent iodine removal performance in aqueous solution demonstrated that the electrostatic attraction between MOF nanosheets with a positive charge and the negatively charged triiodide (I3-, the dominant form of iodine in aqueous solution) is the driving force in adsorption, which endows the adsorbents with the characteristics of fast adsorption and desorption kinetics. The adsorption mechanism was systematically verified by the studies of ζ potential, Fourier transform infrared, X-ray photoelectron spectroscopy, and Raman spectra.

Arousing Electrochemiluminescence Out of Non-Electroluminescent Monomers within Covalent Organic Frameworks.

Covalent organic frameworks (COFs) with stable long-range ordered arrangements are promising materials for organic optoelectronics. However, their electrochemiluminescence (ECL) from non-ECL active monomers has not been realized. Here, we report a design strategy for ECL-emitting COF family. The donors and acceptors co-crystallized and stacked into the highly aligned array of olefin-linked COFs, so that electrons can be transported freely. By this means, a tunable ECL is activated from non-ECL molecules with the maximum efficiency of 32.1% in water with the dissolved oxygen as an inner coreactant, and no additional noxious co-reactant is needed any more. Quantum chemistry calculations further demonstrate that this design reduces the COFs' band gaps and the overlap of electrons and holes in the excited state for better photoelectric properties and stronger ECL signals. This work exploits a basis to envisage the broad application potential of ECL-COFs for various biosensors and light-emitting display.

Deficiency of Histone Methyltransferase SET Domain-Containing 2 in Liver Leads to Abnormal Lipid Metabolism and HCC.

BACKGROUND AND AIMS: Trimethylation of Lys36 on histone 3 (H3K36me3) catalyzed by histone methyltransferase SET domain-containing 2 (SETD2) is one of the most conserved epigenetic marks from yeast to mammals. SETD2 is frequently mutated in multiple cancers and acts as a tumor suppressor. APPROACH AND RESULTS: Here, using a liver-specific Setd2 depletion model, we found that Setd2 deficiency is sufficient to trigger spontaneous HCC. Meanwhile, Setd2 depletion significantly increased tumor and tumor size of a diethylnitrosamine-induced HCC model. The mechanistic study showed that Setd2 suppresses HCC not only through modulating DNA damage response, but also by regulating lipid metabolism in the liver. Setd2 deficiency down-regulated H3K36me3 enrichment and expression of cholesterol efflux genes and caused lipid accumulation. High-fat diet enhanced lipid accumulation and promoted the development of HCC in Setd2-deficient mice. Chromatin immunoprecipitation sequencing analysis further revealed that Setd2 depletion induced c-Jun/activator protein 1 (AP-1) activation in the liver, which was trigged by accumulated lipid. c-Jun acts as an oncogene in HCC and functions through inhibiting p53 in Setd2-deficient cells. CONCLUSIONS: We revealed the roles of Setd2 in HCC and the underlying mechanisms in regulating cholesterol homeostasis and c-Jun/AP-1 signaling.

Tailoring bacterial cellulose structure through CRISPR interference-mediated downregulation of galU in Komagataeibacter xylinus CGMCC 2955.

Diverse applications of bacterial cellulose (BC) have different requirements in terms of its structural characteristics. culturing Komagataeibacter xylinus CGMCC 2955, BC structure changes with alterations in oxygen tension. Here, the K. xylinus CGMCC 2955 transcriptome was analyzed under different oxygen tensions. Transcriptome and genome analysis indicated that BC structure is related to the rate of BC synthesis and cell growth, and galU is an essential gene that controls the carbon metabolic flux between the BC synthesis pathway and the pentose phosphate (PP) pathway. The CRISPR interference (CRISPRi) system was utilized in K. xylinus CGMCC 2955 to control the expression levels of galU. By overexpressing galU and interfering with different sites of galU sequences using CRISPRi, we obtained strains with varying expression levels of galU (3.20-3014.84%). By testing the characteristics of BC, we found that the porosity of BC (range: 62.99-90.66%) was negative with galU expression levels. However, the crystallinity of BC (range: 56.25-85.99%) was positive with galU expression levels; galU expression levels in engineered strains were lower than those in the control strains. Herein, we propose a new method for regulating the structure of BC to provide a theoretical basis for its application in different fields.

SPOP suppresses prostate cancer through regulation of CYCLIN E1 stability.

SPOP is one of the important subunits for CUL3/SPOP/RBX1 complex tightly connected with tumorigenesis. However, its exact roles in different cancers remain debatable. Here, we identify CYCLIN E1, as a novel substrate for SPOP. SPOP directly interacts with CYCLIN E1 and specific regulates its stability in prostate cancer cell lines. SPOP/CUL3/RBX1 complex regulates CYCLIN E1 stability through poly-ubiquitination. CDK2 competes with SPOP for CYCLIN E1 interaction, suggesting that SPOP probably regulates the stability of CDK2-free CYCLIN E1. CYCLIN E1 expression rescued proliferation, migration, and tumor formation of prostate cancer cell suppressed by SPOP. Furthermore, we found SPOP selectively regulates the substrates' stability and signaling pathways in prostate cancer and CCRC cell lines, suggesting that complicated mechanisms exist for SPOP to regulate substrate specificity. Altogether, we have revealed a novel mechanism for SPOP in suppressing prostate cancer and provided evidence to show SPOP has dual functions in prostate cancer and CCRC.

Low birth weight contributed to increased serum IL-6 levels in infantile respiratory syncytial virus infection.

BACKGROUND: To evaluate the role of serum cytokines in the pathogenesis of respiratory syncytial virus (RSV) infection in infants with low birth weight (LBW). METHODS: A prospective observational study was performed, and hospitalized children with lower respiratory tract infection (LRTI) were recruited. Three hundred fifty-eight patients < 1 year met the inclusion criteria: 116 patients had only RSV infection (RSV group); 242 patients had no RSV or other specific pathogen (non-RSV group). Serum interleukin-2 (IL-2), IL-4, IL-6, IL-10, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) were detected through flow cytometry. RESULTS: No significant differences in serum IL-2, 4, 6, 10, and IFN-γ levels were observed between the RSV and non-RSV groups. For RSV infected infants with or without wheezing, delivery mode had no obvious effect on the changes of serum cytokine levels. However, the level of IL-6 in the RSV-infected infants with LBW was significantly higher than that in infants with normal birth weight. CONCLUSIONS: Serum IL-6 level was significantly increased in RSV infected infants with LBW. It is likely that the specific serum cytokine pattern will contribute to our understanding of the pathogenesis of RSV infections, especially in RSV-infected infants with LBW.

Hydroxylation Metabolisms of Crassicauline A in Rats Under Toxic Dose.

BACKGROUND AND OBJECTIVES: Crassicauline A, a C19 diterpenoid alkaloid in Aconitum herbs, is an analgesic drug clinically used in China. The in vivo metabolism of crassicauline A is poorly understood, while potential bioactivation is anticipated via hydroxylation metabolism. This work, therefore, aimed to investigate the in vivo hydroxylation metabolism of crassicauline A in rats. METHODS: Using a de novo developed and validated UPLC-MS/MS method, excretion studies in rats were carried out to investigate the recoveries of crassicauline A and its hydroxylated metabolites in urine and feces. Mass fragmentation analysis was used to identify the detected hydroxylated metabolites. In vitro metabolism assay in liver S9 fraction was employed to preliminarily investigate the inter-species difference of hydroxylation metabolism between rats and human. RESULTS: At a toxic dose of 100 µg/kg, less than 10% and 5% of the administrated dose of crassicauline A were recovered in the urine and feces after single intravenous and oral administration, respectively. Trace of yunaconitine, a possible 3-hydroxylated metabolite of crassicauline A, was detected in urine samples, but not considered to be derived from the in vivo metabolism, because the recovered yunaconitine and crassicauline A was equivalent to their occurrences in the test article. Another hydroxylated metabolite was detected with much higher levels than yunaconitine. Based on chromatographic behaviors and fragmentation analysis, the hydroxylation site of this metabolite was tentatively identified at C-15 on the skeleton, which might have produced a toxic alkaloid known as deoxyjesaconitine. The in vivo observations were consistent with the preliminary in vitro results in liver S9 fraction, in which an inter-species difference was highlighted that rats demonstrated more hydroxylation than human did. CONCLUSIONS: This work disclosed that crassicauline A is elimilated in rats predominantly by metabolism under toxic dosage and the hydroxylation probably at C-15 might be a potential bioactivation pathway in both rats and human.

Preparation and Characterization of Silymarin Synchronized and Sustained Release Dropping Pill.

PURPOSE: This study aimed to develop a synchronized and sustained-release silymarin dropping pill, and to evaluate its pharmacokinetic characteristics. METHOD: Polyoxyethylene stearate, glyceryl monostearate, and stearic acid were used to prepare the dropping pills. X-ray powder diffraction, differential scanning calorimetry, and release were used to evaluate its physicochemical properties. The plasma concentration of silybin in beagle dogs after oral administration of silymarin dropping pills and silymarin capsule was determined by RP-HPLC. RESULTS: Synchronized release was achieved with high similarity factor f2 values between every set of two of the five components. Mean plasma concentration-time curves of silymarin after oral administration of dropping pills in beagle dogs were in accordance with first-order absorption and open twocompartment model. The Tmax, Cmax, and AUC0-∞ of dropping pills in beagle dogs were 0.8750±0.13 h, 0.8183±0.07 µg·ml-1, and 2.274±0.90 µg·h·ml-1, respectively. Silymarin dropping pills prolonged in vivo exposure and reduced maximum in vivo concentration, achieving a stable level in the serum. CONCLUSION: The combination of solid dispersion technique and dropping pill formulation allowed synchronized release of multiple components in herbal medicine, and has potential application in the development of sustained release in herbal medicine.

TTLL12 Inhibits the Activation of Cellular Antiviral Signaling through Interaction with VISA/MAVS.

Upon virus infection, host cells use retinoic-acid-inducible geneI I (RIG-I)-like receptors to recognize viral RNA and activate type I IFN expression. To investigate the role of protein methylation in the antiviral signaling pathway, we screened all the SET domain-containing proteins and identified TTLL12 as a negative regulator of RIG-I signaling. TTLL12 contains SET and TTL domains, which are predicted to have lysine methyltransferase and tubulin tyrosine ligase activities, respectively. Exogenous expression of TTLL12 represses IFN-ß expression induced by Sendai virus. TTLL12 deficiency by RNA interference and CRISPR-gRNA techniques increases the induced IFN-ß expression and inhibits virus replication in the cell. The global gene expression profiling indicated that TTLL12 specifically inhibits the expression of the downstream genes of innate immunity pathways. Cell fractionation and fluorescent staining indicated that TTLL12 is localized in the cytosol. The mutagenesis study suggested that TTLL12's ability to repress the RIG-I pathway is probably not dependent on protein modifications. Instead, TTLL12 directly interacts with virus-induced signaling adaptor (VISA), TBK1, and IKKε, and inhibits the interactions of VISA with other signaling molecules. Taken together, our findings demonstrate TTLL12 as a negative regulator of RNA-virus-induced type I IFN expression by inhibiting the interaction of VISA with other proteins.

Serum cytokine profile contributes to discriminating M. pneumoniae pneumonia in children.

BACKGROUND: To evaluate the role of serum cytokines in discriminating M. pneumoniae infection in children with community-acquired pneumonia (CAP). METHODS: A prospective observational study was conducted. 385 hospitalized patients with CAP had only M. pneumoniae (MP group) infection; 321 hospitalized patients with CAP had no M. pneumoniae and other specific pathogen (control group) infections. Serum interleukin-2 (IL-2), IL-4, IL-6, IL-10, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) were detected by flow cytometry. RESULTS: In children younger than 5years, serum IL-6, TNF-α, and IFN-γ levels from MP group were significantly higher than those from control group. However in children 5-15years, serum IL-6, IL-10, and IFN-γ levels from MP group were significantly higher than those from control group. In the final multivariate logistic regression model for serum cytokine, moderately elevated IL-6, IL-10, and IFN-γ shows a higher prediction of development of M. pneumoniae pneumonia among CAP patients. CONCLUSIONS: A specific cytokine pattern showed a higher prediction of M. pneumoniae pneumonia among CAP patients, further suggesting that serum cytokine pattern might be useful in differentiating infectious causative agents in children.

A chemical profiling strategy for semi-quantitative analysis of flavonoids in Ginkgo extracts.

Flavonoids analysis in herbal products is challenged by their vast chemical diversity. This work aimed to develop a chemical profiling strategy for the semi-quantification of flavonoids using extracts of Ginkgo biloba L. (EGB) as an example. The strategy was based on the principle that flavonoids in EGB have an almost equivalent molecular absorption coefficient at a fixed wavelength. As a result, the molecular-contents of flavonoids were able to be semi-quantitatively determined by the molecular-concentration calibration curves of common standards and recalculated as the mass-contents with the characterized molecular weight (MW). Twenty batches of EGB were subjected to HPLC-UV/DAD/MS fingerprinting analysis to test the feasibility and reliability of this strategy. The flavonoid peaks were distinguished from the other peaks with principle component analysis and Pearson correlation analysis of the normalized UV spectrometric dataset. Each flavonoid peak was subsequently tentatively identified by the MS data to ascertain their MW. It was highlighted that the flavonoids absorption at Band-II (240-280 nm) was more suitable for the semi-quantification purpose because of the less variation compared to that at Band-I (300-380 nm). The semi-quantification was therefore conducted at 254 nm. Beyond the qualitative comparison results acquired by common chemical profiling techniques, the semi-quantitative approach presented the detailed compositional information of flavonoids in EGB and demonstrated how the adulteration of one batch was achieved. The developed strategy was believed to be useful for the advanced analysis of herbal extracts with a high flavonoid content without laborious identification and isolation of individual components.

Highly efficient electrochemiluminescence based on pyrazolecarboxylic metal organic framework.

A series of transition metal complexes with the ligands 3-pyrazoledicarboxylic acid (H2L(1)) and ethyl 1-(2-ethoxy-2-oxoethyl)-1H-pyrazole-3-carboxylate (epzc) have been synthesized. The epzc generated 1-(carboxymethyl)-1H-pyrazole-3-carboxylic acid (H2L(2)) by an in situ hydrothermal hydrolysis reaction, using a one-pot method. Simple mononuclear [Co(HL(1))2(H2O)2] (1) and [Ni(L(2))(H2O)4] (4), dinuclear [Ni2(L(1))2(H2O)6]·H2O (2) and [Cu2(L(2))2(H2O)4] (5) and 2D frameworks [Cu2(L(1))2]n (3), [Co2(L(2))2(H2O)4]n (6) have been isolated. The structures have been established by single-crystal X-ray diffraction, and the complexes characterized by FT-IR, thermogravimetric analysis (TGA), PRXD, UV-Vis spectroscopy, and fluorescent spectroscopy. 1, 2, 4 and 4 were further assembled to form a supramolecular structure by hydrogen-bonding interactions and/or π···π stacking. 3 and 6 both possess a 2D network structure that is further interlinked via intermolecular hydrogen-bonding interactions. Most importantly, the complexes demonstrated highly intense electrochemiluminescence (ECL) in DMF solution.

Characterizations of the hydrolyzed products of ginkgolide A and ginkgolide B by liquid chromatography coupled with mass spectrometry.

Ginkgolides are diterpenoid trilactones responsible for the neuromodulatory properties of Ginkgo biloba extracts. They are to be hydrolyzed in aqueous solutions as mixed carboxylate forms potentially including three monocarboxylates, three dicarboxylates and one tricarboxylate. Characterizations of the hydrolyzed products are challenging because there is no way to prepare them individually. In this work, the major hydrolyzed products of ginkgolide A (GA) and ginkgolide B (GB) including all three monocarboxyaltes have been produced in buffers and subjected to liquid chromatography coupled with triple quadrupole MS and LTQ Orbitrap MS analysis. With the comparative analysis of the trilactone of GA and GB, it was highlighted a unique charge-driven fragmentation pathway of twice neutral losses of CO on the lactone-C. The monocarboxylates were accordingly identified based on the construction of their fragmentation pathways cross-linked with those of the trilactone. In brief, the lactone-C hydrolyzed product is characteristic of the absence of product ions between [M-H](-) and [M-H-C2H2O3](-) (m/z 351 for GA and m/z 367 for GB). The featured fragmentation pathway of the lactone-F hydrolyzed product is the cleavage of ring-A, yielding a fragment (m/z 295 for GA and m/z 309 for GB) followed with twice (GA) or triple (GB) neutral losses of CO. The most characteristic fragment of the lactone-E hydrolyzed product is [M-H-H2O-CO2-2CO](-) (m/z 307 for GA and m/z 323 for GB) in contrast to the other two monocarboxylates. The knowledge gained in this work was of special uses to investigate the biological fates and the corresponding pharmacological mechanisms of ginkgolides.

Understanding the regioselective hydrolysis of ginkgolide B under physiological environment based on generation, detection, identification, and semi-quantification of the hydrolyzed products.

A liquid chromatography-mass spectrometry (LC-MS) method coupled with specialized sample-preparation strategies was developed to investigate the hydrolysis of ginkgolide B (GB) in physiological environments in comparison with that of ginkgolide A (GA). The rapid hydrolysis processes were captured by the direct injection of samples prepared in the volatile buffers. The LC-MS behavior of the hydrolyzed products, including three monocarboxylates and three dicarboxylates, was acquired. The monocarboxylates were identified by fragmentation analysis, and the dicarboxylates were accordingly tentatively identified by reaction sequences. The base-catalyzed hydrolysis of GB and GA was characterized at 4 °C within pH 7.0-10.7. The regioselective reactions on the lactone-C and lactone-F were revealed by thermodynamic studies at pH 6.8 and 7.4. It was revealed that the 1-hydroxyl group on the skeleton of GB blocks the reactivity of the lactone-E. On the basis of these results, a distinctive hydrolysis phenomenon of GB was confirmed in plasma of humans, rats, and dogs as a rapid degradation of the trilactone along with the only production of the lactone-F-hydrolyzed product. This phenomenon is also closely associated with the 1-hydroxyl group, because it was not observed in GA. More interestingly, the underlying mechanism was revealed not to be associated with any typical enzyme-catalyzed process, but to be potentially involved with a selective reaction of the intact or broken lactone-C moiety with endogenous small-molecule reactants in plasma. This in-depth knowledge of the hydrolysis of GB versus GA not only facilitated understanding of their pharmacological mechanisms but also provided potential routes to study the structure-activity relationships of ginkgolides. Graphical Abstract Regioselective hydrolysis of ginkgolide B in pH 7.4 buffers and plasma.

Semi-quantitative determination of monocarboxylate forms of ginkgolide B in plasma by UPLC-MS.

Ginkgolide B (GB) has a unique structure incorporating three γ-lactones that may be hydrolyzed in aquae as carboxylate forms. However, the determinations of them are challenging because there is no way to prepare the standards of any hydrolyzed products of GB in the solid state. In this report, a semi-quantitative method was developed to determine the monocarboxylate forms of GB in plasma. UPLC coupled with selected ion monitoring (SIR) of m/z 423 and m/z 441 were employed to detect the trilactone and monocarboxylates in assistances with the frozen method and the recovered method, which were, respectively, used to stabilize the hydrolyzed states and fully recover the monocarboxylates as the trilactone in samples. Two monocarboxylates were detected in pH 7.4 potassium phosphate buffers (PPB) after incubations, while only one was found in plasma in vitro and in vivo. The identifications of them require further studies. Following the bioanalytical validation of the trilactone, critical issues of the relative responses of the monocarboxylates in contrast to the trilactone, matrix effects, and stabilities were carefully investigated with subtly designed measures. The validation results supported the quantifications of monocarboxylates in PPB or plasma directly by using the corresponding calibration curve of the trilactone. The applications of this method presented a clear disparity between the hydrolysis kinetics of GB in plasma and PPB. Based on the quantification results and method applications, it was concluded that the present method was suitable to study the complex hydrolysis mechanisms of GB in vitro and in vivo.