Ubiquitinome Analysis Uncovers Alterations in Synaptic Proteins and Glucose Metabolism Enzymes in the Hippocampi of Adolescent Mice Following Cold Exposure.

Cold exposure exerts negative effects on hippocampal nerve development in adolescent mice, but the underlying mechanisms are not fully understood. Given that ubiquitination is essential for neurodevelopmental processes, we attempted to investigate the effects of cold exposure on the hippocampus from the perspective of ubiquitination. By conducting a ubiquitinome analysis, we found that cold exposure caused changes in the ubiquitination levels of a variety of synaptic-associated proteins. We validated changes in postsynaptic density-95 (PSD-95) ubiquitination levels by immunoprecipitation, revealing reductions in both the K48 and K63 polyubiquitination levels of PSD-95. Golgi staining further demonstrated that cold exposure decreased the dendritic-spine density in the CA1 and CA3 regions of the hippocampus. Additionally, bioinformatics analysis revealed that differentially ubiquitinated proteins were enriched in the glycolytic, hypoxia-inducible factor-1 (HIF-1), and 5'-monophosphate (AMP)-activated protein kinase (AMPK) pathways. Protein expression analysis confirmed that cold exposure activated the mammalian target of rapamycin (mTOR)/HIF-1α pathway. We also observed suppression of pyruvate kinase M2 (PKM2) protein levels and the pyruvate kinase (PK) activity induced by cold exposure. Regarding oxidative phosphorylation, a dramatic decrease in mitochondrial respiratory-complex I activity was observed, along with reduced gene expression of the key subunits NADH: ubiquinone oxidoreductase core subunit V1 (Ndufv1) and Ndufv2. In summary, cold exposure negatively affects hippocampal neurodevelopment and causes abnormalities in energy homeostasis within the hippocampus.

Metabolic Regulation-Mediated Reversion of the Tumor Immunosuppressive Microenvironment for Potentiating Cooperative Metabolic Therapy and Immunotherapy.

Tumor cells exhibit heightened glucose (Glu) consumption and increased lactic acid (LA) production, resulting in the formation of an immunosuppressive tumor microenvironment (TME) that facilitates malignant proliferation and metastasis. In this study, we meticulously engineer an antitumor nanoplatform, denoted as ZLGCR, by incorporating glucose oxidase, LA oxidase, and CpG oligodeoxynucleotide into zeolitic imidazolate framework-8 that is camouflaged with a red blood cell membrane. Significantly, ZLGCR-mediated consumption of Glu and LA not only amplifies the effectiveness of metabolic therapy but also reverses the immunosuppressive TME, thereby enhancing the therapeutic outcomes of CpG-mediated antitumor immunotherapy. It is particularly important that the synergistic effect of metabolic therapy and immunotherapy is further augmented when combined with immune checkpoint blockade therapy. Consequently, this engineered antitumor nanoplatform will achieve a cooperative tumor-suppressive outcome through the modulation of metabolism and immune responses within the TME.

Bioorthogonal "Click and Release" Reaction-Triggered Aggregation of Gold Nanoparticles Combined with Released Lonidamine for Enhanced Cancer Photothermal Therapy.

Cancer has been the most deadly disease, and 13 million cancer casualties are estimated to occur each year by 2030. Gold nanoparticles (AuNPs)-based photothermal therapy (PTT) has attracted great interest due to its high spatiotemporal controllability and noninvasiveness. Due to the trade-off between particle size and photothermal efficiency of AuNPs, rational design is needed to realize aggregation of AuNPs into larger particles with desirable NIR adsorption in tumor site. Exploiting the bioorthogonal "Click and Release" (BCR) reaction between iminosydnone and cycloalkyne, aggregation of AuNPs can be achieved and attractively accompanied by the release of chemotherapeutic drug purposed to photothermal synergizing. We synthesize iminosydnone-lonidamine (ImLND) as a prodrug and choose dibenzocyclooctyne (DBCO) as the trigger of BCR reaction. A PEGylated AuNPs-based two-component nanoplatform consisting of prodrug-loaded AuNPs-ImLND and tumor-targeting peptide RGD-conjugated AuNPs-DBCO-RGD is designed. In the therapeutic regimen, AuNPs-DBCO-RGD are intravenously injected first for tumor-specific enrichment and retention. Once the arrival of AuNPs-ImLND injected later at tumor site, highly photothermally active nanoaggregates of AuNPs are formed via the BCR reaction between ImLND and DBCO. The simultaneous release of lonidamine further enhanced the therapeutic performance by sensitizing cancer cells to PTT.

CRMP5 participates in oocyte meiosis by regulating spastin to correct microtubule-kinetochore misconnection.

Our previous studies have suggested that spastin, which aggregates on spindle microtubules in oocytes, may promote the assembly of mouse oocyte spindles by cutting microtubules. This action may be related to CRMP5, as knocking down CRMP5 results in reduced spindle microtubule density and maturation defects in oocytes. In this study, we found that, after knocking down CRMP5 in oocytes, spastin distribution shifted from the spindle to the spindle poles and errors in microtubule-kinetochore attachment appeared in oocyte spindles. However, CRMP5 did not interact with the other two microtubule-severing proteins, katanin-like-1 (KATNAL1) and fidgetin-like-1 (FIGNL1), which aggregate at the spindle poles. We speculate that, in oocytes, due to the reduction of spastin distribution on chromosomes after knocking down CRMP5, microtubule-kinetochore errors cannot be corrected through severing, resulting in meiotic division abnormalities and maturation defects in oocytes. This finding provides new insights into the regulatory mechanisms of spastin in oocytes and important opportunities for the study of meiotic division mechanisms.

Light-driven biohybrid system utilizes N2 for photochemical CO2 reduction.

Attempting to couple photochemical CO2 reduction with N2 fixation is usually difficult, because the reaction conditions for these two processes are typically incompatible. Here, we report that a light-driven biohybrid system can utilize abundant, atmospheric N2 to produce electron donors via biological nitrogen fixation, to achieve effective photochemical CO2 reduction. This biohybrid system is constructed by incorporating molecular cobalt-based photocatalysts into N2-fixing bacteria. It is found that N2-fixing bacteria can convert N2 into reductive organic nitrogen and create a localized anaerobic environment, which allows the incorporated photocatalysts to continuously perform photocatalytic CO2 reduction under aerobic conditions. Specifically, the light-driven biohybrid system displays a high formic acid production rate of over 1.41 × 10-14 mol h-1 cell-1 under visible light irradiation, and the organic nitrogen content undergoes an over-3-fold increase within 48 hours. This work offers a useful strategy for coupling CO2 conversion with N2 fixation under mild and environmentally benign conditions.

Integrated cascade catalysis of microalgal bioenzyme and inorganic nanozyme for anti-inflammation therapy.

Combinations of multiple enzymes for cascade catalysis have been widely applied in biomedicine, but the integration of a natural bioenzyme with an inorganic nanozyme is less developed. Inspired by the abundant content of superoxide dismutase (SOD) in Spirulina platensis (SP), we establish an integrated cascade catalysis for anti-inflammation therapy by decorating catalase (CAT)-biomimetic ceria nanoparticles (CeO2) onto the SP surface via electrostatic interaction to build microalgae-based biohybrids. The biohybrids exhibit combined catalytical competence for preferentially transforming superoxide anion radicals (O2˙-) to hydrogen peroxide (H2O2), and subsequently catalyzing H2O2 disproportionation to water and oxygen. In ulcerative colitis and Crohn's disease, the biohybrids reveal a satisfactory therapeutic effect owing to the synergistic reactive oxygen species (ROS)-scavenging capacity, suggesting a new train of thought for enzyme-based biomedical application.

Hierarchy-Assembled Dual Probiotics System Ameliorates Cholestatic Drug-Induced Liver Injury via Gut-Liver Axis Modulation.

Cholestatic drug-induced liver injury (DILI) induced by drugs or other xenobiotics is a severe and even fatal clinical syndrome. Here, living materials of hierarchy-assembled dual probiotics system are fabricated by sequentially encapsulating probiotic Lactobacillus delbrueckii subsp. bulgaricus (LDB) and Lactobacillus rhamnosus GG (LGG) into Ca2+ -complexed polymer microspheres for effective prevention of cholestatic DILI. Upon entering intestinal tract of the constructed living materials, LGG is released because of pH-triggered dissolution of outer enteric polymer coating. The released LGG can inhibit hepatic bile acids (BAs) synthesis by activating intestinal farnesoid X receptor-fibroblast growth factor 15（FGF-15) signaling pathway. BAs excretion is also facilitated by LGG through increasing the abundance of bile salt hydrolase (BSH)-active gut commensal bacteria. Furthermore, exposed positively-charged chitosan shell can absorb the excessive BAs via electrostatic interaction, which leads to steady BAs fixation by the imprisoned LDB, decreasing the total BAs amounts in enterohepatic circulation. Together, the fabricated living materials, obtained here, can effectively prevent cholestatic DILI through dredging cholestasis via gut-liver axis modulation. The therapeutic effect is demonstrated in α-naphthylisothiocyanate and clinical antiepileptic drug valproate acid-induced cholestatic DILI mouse models, which reveal the great potential for effective cholestatic DILI management.

Development and evaluation of short form of constitution in Chinese medicine questionnaire: a national epidemiological survey data of 21 948 case.

OBJECTIVE: To develop the best short form of constitution in Chinese medicine questionnaire (CCMQ) and evaluate its psychometric properties in Chinese population. METHODS: A total of 21 948 subjects were used to refine the short form. Correlation coefficient, exploratory factor analysis (EFA) and Cronbach's alpha coefficient were used to analyze and select items to form the short form. Separate sample of 205 subjects were collected to further evaluate the short from. EFA, confirmatory factor analysis (CFA), item-scale correlation, discriminant validity, internal consistency reliability and split-half reliability were carried out to evaluate the short form. RESULTS: The short form CCMQ included 26 items. Seven common factors of characteristic root > 1 were extracted to explain 58.488% of the total variation. Result of CFA was consistent with the 9-factors structure. The mean differences of Blood-stasis body constitution and Qi-stagnation body constitution had statistical significance in body mass index differentiation. Cronbach's alpha coefficient of short form CCMQ was 0.863. The split-half reliability of total scale was 0.813, and each scale was 0.568-0.770. The item-scale correlations ranged from 0.620-0.849. CONCLUSION: The short form CCMQ consisted of 26 items with good psychometric properties. The short form should be recommended for the measurement of health of Chinese population in any clinical trial.

Large π-Conjugated Metal-Organic Frameworks for Infrared-Light-Driven CO2 Reduction.

It remains challenging to excite traditional photocatalysts through near-infrared (NIR) light. Attempts to use NIR-light-response materials for photochemical reduction usually suffer from inapposite band position due to extremely narrow band gaps. Here, we report that large π-conjugated organic semiconductor engineered metal-organic framework (MOF) can result in NIR-light-driven CO2 reduction catalyst with high photocatalytic activity. A series of mesoporous MOFs, with progressively increased macrocyclic π-conjugated units, were synthesized for tuning the light adsorption range and catalytic performance. Attainment of these MOFs in single-crystal form revealed the identical topology and precise spatial arrangements of constituent organic semiconductor units and metal clusters. Furthermore, the ultrafast spectroscopic studies confirmed the formation of charge separation state and the mechanism underlying photoexcited dynamics. This combined with X-ray photoelectron spectroscopy and in situ electron paramagnetic resonance studies verified the photoinduced electron transfer pathway within MOFs for NIR-light-driven CO2 reduction. Specifically, tetrakis(4-carboxybiphenyl)naphthoporphyrin) MOF (TNP-MOF) photocatalyst displayed an unprecedentedly high CO2 reduction rate of over 6630 µmol h-1 g-1 under NIR light irradiation, and apparent quantum efficiencies (AQE) at 760 and 808 nm were over 2.03% and 1.11%, respectively. The photocatalytic performance outperformed all the other MOF-based photocatalysts, even visible-light-driven MOF-based catalysts.

Highly Stable Iron Carbonyl Complex Delivery Nanosystem for Improving Cancer Therapy.

Metal carbonyl complexes can readily liberate carbon monoxide (CO) in response to activation stimulus. However, applicability of metal carbonyl complexes is limited because they are unstable under natural ambient conditions of moisture and oxygen. Reported here is the rational design of an iron carbonyl complex delivery nanosystem for the improvement of cancer therapy. We demonstrated that iron pentacarbonyl (Fe(CO)5) can be encapsulated into the cavity of a Au nanocage under an oxygen-free atmosphere and then controllably form iron oxide on the surface of the Au nanocage under aerobic conditions. The formation of iron oxide efficiently avoids the leakage and oxidation of the caged Fe(CO)5. The resulting nanomaterial exhibits excellent safety, biocompatibility, and stability, which can be specifically activated under near-infrared (NIR) irradiation within the tumor environment to generate CO and iron. The released CO causes damage to mitochondria and subsequent initiation of autophagy. More importantly, during autophagy, the nanomaterial that contains iron and iron oxide can accumulate into the autolysosome and result in its destruction. The produced CO and iron show excellent synergistic effects in cancer cells.

The development process of self-acceptance among Chinese women with breast cancer.

AIM: The development process of self-acceptance in breast cancer survivors is a dynamic process that is poorly understood. The objective of the present study was to explore and delineate the dynamic progression toward self-acceptance in Chinese women with breast cancer. METHODS: Data were collected through individual in-depth face-to-face interviews with 20 women who had undergone treatment for breast cancer at the breast center in a large tertiary care hospital in Ningbo, China between September 2016 and June 2017. Data analysis occurred through the open, axial, and selective coding stages of grounded theory and used the constant comparative method. RESULTS: Based on the interviewer responses, one core category, three categories, and seven subcategories were identified that pertained to the process of self-acceptance in Chinese women with breast cancer. The core category of self-acceptance was normalization, returning to the pre-illness state with an identity and image that conformed to the cultural norm. To reach normalization, women progressed through a crisis stage, a compromise stage, and a managing impressions stage. CONCLUSION: This study proposes that self-acceptance in breast cancer survivors is a dynamic and active process. Findings will inform the development of interventions that will provide structure and support to Chinese women with breast cancer.

AMPK regulates anaphase central spindle length by phosphorylation of KIF4A.

AMP-activated protein kinase (AMPK) is an energy sensor that couples the cellular energy state with basic biological processes. AMPK is thought to be linked with cell division although the underlying mechanisms remain largely unknown. Here, we show that AMPK functionally participates throughout cell division and that AMPK catalytic subunits, especially α2, are sequentially associated with separate mitotic apparatus. Using quantitative phosphoproteomics analysis, we found that the strong direct substrate KIF4A is phosphorylated by AMPK at Ser801. Further analysis revealed that AMPK and Aurora B competitively phosphoregulates KIF4A during mitotic phase due to overlapping recognition motifs, resulting in the elaborate phosphoregulation for KIF4A-dependent central spindle length control. Given the intrinsic energy-sensing function of AMPK, our study links the KIF4A-dependent control of central spindle length with cellular glucose stress.

Graphene oxide-enhanced cytoskeleton imaging and mitosis tracking.

Here we show that graphene oxide greatly enhances the imaging ability of a peptide probe that selectively targets microtubules of the cytoskeleton, thus enabling the dynamic tracking of mitosis in live cells.

Protein kinase A rescues microtubule affinity-regulating kinase 2-induced microtubule instability and neurite disruption by phosphorylating serine 409.

Microtubule affinity-regulating kinase 2 (MARK2)/PAR-1b and protein kinase A (PKA) are both involved in the regulation of microtubule stability and neurite outgrowth, but whether a direct cross-talk exists between them remains unclear. Here, we found the disruption of microtubule and neurite outgrowth induced by MARK2 overexpression was blocked by active PKA. The interaction between PKA and MARK2 was confirmed by coimmunoprecipitation and immunocytochemistry both in vitro and in vivo. PKA was found to inhibit MARK2 kinase activity by phosphorylating a novel site, serine 409. PKA could not reverse the microtubule disruption effect induced by a serine 409 to alanine (Ala) mutant of MARK2 (MARK2 S409A). In contrast, mutation of MARK2 serine 409 to glutamic acid (Glu) (MARK2 S409E) did not affect microtubule stability and neurite outgrowth. We propose that PKA functions as an upstream inhibitor of MARK2 in regulating microtubule stability and neurite outgrowth by directly interacting and phosphorylating MARK2.

Influence of human myasthenia gravis thymus on the differentiation of human cord blood stem cells in SCID mice.

The normal thymus contributes to T lymphocytes differentiation and induction of tolerance to self-antigens. Myasthenia gravis (MG) is characterized by abnormal thymic hyperplasia. To assess the potential influence of MG-thymus on the differentiation of T lymphocytes differentiation, we used the MG-thymus transplanted severe combined immunodeficiency (SCID) mice model to evaluate the human cord blood stem cells differentiation. Thymus fragments from MG patient and human cord blood stem cells were transplanted into SCID mice successively. SCID mice were observed to develop sustained human T lymphocytes and a functional anti-tumor immune. The levels of various T cell subsets in SCID mice with MG-thymus were different from that of control group. Among that, the frequency of CD4(+)CD25(+) T cells was significant lower in SCID mice with MG-thymus. The deficiency of CD4(+)CD25(+) T cells seens to contribute to the pathogenesis of MG.

[Effects of CD74 gene on IFNγR gene expression in MG TEC].

AIM: To study the effects of CD74 gene on IFN-γR expression in myasthenia gravis thymic epithelial cell(TEC). METHODS: PCMV-CD74 transiently transfect to primary cultured TEC mediated with lipofectamine, the result of transfection and some autoimmune related genes such as IFN-γR were detected by real-time RT-PCR. RESULTS: Real-time quantitative RT-PCR results show that the mRNA expression level of IL-4R, IL-32, IFN-γR, ECGF1, HLA-A, HLA-DR increased significantly, with the high express ion of CD74 gene in thymic epithelial cells. CONCLUSION: CD74 gene over-expression in TEC can increase IFN-γR mRNA expression.

[Changes of differentiation and gene expression of CD133+ cells in human umbilical cord blood induced by SCF and bFGF].

This study was aimed to investigate the changes of differentiation and gene expression of CD133(+) cells in human umbilical cord blood induced by stem cell factor (SCF) and basic fibroblast growth factor (bFGF) in vitro, and to explore the biological characteristics of CD133(+) cells so as to provide experimental basis for potential use in regenerative medicine. The human umbilical cord blood CD133(+) cells were isolated from umbilical cord blood and purified by MACS magnetic selection. The CD133(+) cells were cultured in DMEM/F12 medium containing SCF, bFGF and B27 for 10 to 15 days. The total RNA of these cells was extracted before and after culture, and the analysis of related gene expression levels of these cells was performed using oligonucleotide microarrays. The results showed that out of 263 genes 21 genes were obviously up-regulated after culture than that before culture, whereas 7 genes were found to be significant down-regulated as compared with fresh-separated CD133(+) cells. These genes were involved in stem cell specific markers, cell cycle regulators, stem cell differentiation markers and signaling pathways that are important for stem cell maintenance. It is concluded that SCF and bFGF can induce differentiation of human cord blood CD133(+) cells through up- or down-regulation of specific genes. This study provides gene expression information for SCF and bFGF-induced human cord blood CD133(+) cells and contributes to understand the effect of SCF and bFGF on proliferation and differentiation CD133(+) cells at gene level, and promotes therapeutic applications of the CD133(+) cells induced by SCF and bFGF.

Toxicity of derivatives from semicarbazide-sensitive amine oxidase-mediated deamination of methylamine against Toxoplasma gondii after infection of differentiated 3T3-L1 cells.

Adipose tissue plays an active role in normal metabolic homeostasis as well as in the development of human diseases such as atherosclerosis and diabetes. We report here antimicrobial activities of the metabolites from adipocytes. Specifically, semicarbazide-sensitive amine oxidase of differentiated 3T3-L1 cells was found to utilize methylamine for producing formaldehyde and hydrogen peroxide, accounting for the inhibition of infectivity of Toxoplasma gondii and its replication in these cells. This was demonstrated by the findings that semicarbazide-sensitive amine oxidase was extremely high in differentiated 3T3-L1 cells; and that the infection of these cells by T. gondii and its intracellular replication were decreased to 33% and 37% of the control, respectively, when methylamine was provided in micromolar concentrations as the substrate to the aminoxidase. Only one of the two reaction products expected was found inhibitory against T. gondii when added to the infected pre-adipocytes of 3T3-L1. Intracellular replication of this parasite was inhibited by formaldehyde in the range of 10-100 microM and stimulated by hydrogen peroxide at 1-10 microM. The finding indicates that T. gondii may be useful as a sensitive and convenient sentinel for screening agents toxic to eukaryotic cells.

Edgetic perturbation models of human inherited disorders.

Cellular functions are mediated through complex systems of macromolecules and metabolites linked through biochemical and physical interactions, represented in interactome models as 'nodes' and 'edges', respectively. Better understanding of genotype-to-phenotype relationships in human disease will require modeling of how disease-causing mutations affect systems or interactome properties. Here we investigate how perturbations of interactome networks may differ between complete loss of gene products ('node removal') and interaction-specific or edge-specific ('edgetic') alterations. Global computational analyses of approximately 50,000 known causative mutations in human Mendelian disorders revealed clear separations of mutations probably corresponding to those of node removal versus edgetic perturbations. Experimental characterization of mutant alleles in various disorders identified diverse edgetic interaction profiles of mutant proteins, which correlated with distinct structural properties of disease proteins and disease mechanisms. Edgetic perturbations seem to confer distinct functional consequences from node removal because a large fraction of cases in which a single gene is linked to multiple disorders can be modeled by distinguishing edgetic network perturbations. Edgetic network perturbation models might improve both the understanding of dissemination of disease alleles in human populations and the development of molecular therapeutic strategies.

[Expression of CD45RA and CD45RO thymocyte subset and related cytokine in patients with myasthenia gravis].

AIM: To study the relationship between the abnormal differentiation thymocytes and the pathogenesis of myasthenia gravis (MG). METHODS: The gene expression of some ILs, IFN and their receptors was examined by cDNA array. The percentage of CD45RO(+) and D45RA(+) thymocytes was measured by flow cytometry (FCM). The expression and distribution of CD45RA and CD45RO in the samples of thymus tissues were observed by immunohistochemistry. RESULTS: The levels of IL-1R, IL-4R, IFNgammaR1, IFNgammaR2, IL-6 and IL-8 in the patients with MG were lower than those of normal controls and there were significant differences between them. But the levels of IL-1, IL-2, IL-4, IL-10, IL-7, IFN-alpha, IFN-beta and IFN-gamma showed no significant changes in patients with MG in comparison with normal controls. The percentage of CD56(+) thymocytes in MG patients 0.56+/-0.33 was significantly lower than that in normal controls(1.78+/-0.69).The CD45RO(+) and CD1a(+) in MG patients increased more significantly those in normal controls. The immunohistochemistry and RT-PCR showed the same result. CONCLUSION: Aberrant transformation from CD45R0(+)T to CD45RA(+)T has been found in the development of thymocytes of the patients with MG.