Hirschsprung's disease: m6A methylase VIRMA suppresses cell migration and proliferation by regulating GSK3ß.

BACKGROUND: N6-methyladenosine (m6A) is the most abundant mRNA modification in mammals, participating in various biological processes. VIRMA is a key methyltransferase involved in m6A modification. However, the role of VIRMA in Hirschsprung's disease (HSCR) remains unclear. This study aims to investigate the function of VIRMA in HSCR and identify its corresponding regulatory mechanisms. METHODS: The expression of VIRMA and GSK3ß in colon tissues of HSCR was examined using RT-qPCR, Western blot, and Immunohistochemistry. Immunofluorescence detected localization of VIRMA and GSK3ß. Cell proliferation was measured by CCK8 and EdU assays, and cell migration was evaluated via cell migration and wound healing assays. The stability of GSK3ß mRNA was assessed using the actinomycin D assay and the overall level of m6A in cells was assessed by colorimetric assay. RESULTS: VIRMA was significantly downregulated in narrow-segment colon tissue. Silencing of VIRMA inhibited cell proliferation and migration. VIRMA can inhibit the degradation of GSK3ß mRNA and increase the expression of GSK3ß. GSK3ß was significantly upregulated in narrow-segment colon tissues. Accordingly, our findings showed that GSK3ß mediated the VIRMA-driven cell migration and proliferation. CONCLUSION: VIRMA can inhibit cell migration and proliferation by upregulating the expression of GSK3ß, contributing to the onset of HSCR. IMPACT: The expressions of VIRMA were significantly reduced in HSCR, while GSK3ß expression was increased in HSCR, and can be used as a molecular marker. VIRMA overexpression promoted the proliferation and migration of SH-SY5Y and HEK-293T cells. VIRMA can inhibit the degradation of GSK3ß mRNA and increase the expression of GSK3ß.

Characterization of beta subunit variants of recombinant human chorionic gonadotrophin.

Human chorionic gonadotropin (hCG), an endogenous glycoprotein hormone, has been widely used for the treatment of infertility and corpus luteum defect in women. The biological specificity of hCG is essentially determined by its beta (ß-) subunit, whereas the alpha (α-) subunit is a common subunit shared among the gonadotropin family. In development of a therapeutic recombinant hCG, the purity analysis showed that the beta (ß-) subunit has two variants, ß1 and ß2. Structural characterization using a combination of analytical techniques has demonstrated that ß1-subunit is derived from non-glycosylation at Asn 13, whereas ß2-subunit is a normal species with complete N-glycosylation at both Asn 13 and Asn 30. In vivo Bioactivity evaluation of the r-hCG fractions with various ratios of ß1-and ß2-subunits showed that incomplete glycosylation at Asn 13 potentially reduced the biological activity of r-hCG to promote uterus growth. Although hCG has a long history of medicinal use, this is the first report to identify the structural difference of hCG ß-subunit variants, as well as to preliminary establish the structure-activity relationship of this variation. The obtained results also suggest the importance of variant characterization and necessary quality control of product variants during the development of recombinant protein therapeutics.

Cu-Catalyzed Decarboxylative Annulation of N-Phenylglycines with Maleimides: Synthesis of 1H-Pyrrolo[3,4-c]quinoline-1,3(2H)-diones.

A novel protocol for the construction of functionalized 1H-pyrrolo[3,4-c]quinoline-1,3(2H)-diones (PQLs, 3) from N-phenylglycines and maleimides was developed. The cascade reaction was enabled by heating a mixture of the two substrates in the presence of di-tert-butyl peroxide (DTBP) as an oxidant and anhydrous CuBr as a catalyst in chlorobenzene. Consequently, a diverse series of PQLs 3 were synthesized in moderate-to-good yields (43-73%). The synthesis of the PQLs was enabled via a one-pot cascade reaction that proceeded through subsequent oxidative decarboxylation, 1,2-addition, intramolecular cyclization, tautomerization, and aromatization reactions. This protocol can be used for the synthesis of functionalized PQLs via a one-pot oxidative decarboxylation annulation reaction rather than through a series of multistep reactions, making it suitable for both combinatorial and parallel syntheses of PQLs.

Effect of pelvic floor muscle training on urinary incontinence after radical prostatectomy: An umbrella review of meta-analysis and systematic review.

OBJECTIVE: To analyse the specific exercise effects of pelvic floor muscle training (PFMT) with or without biofeedback or electrical stimulation on urinary incontinence rehabilitation after radical prostatectomy. DATA SOURCES: We searched PubMed, Embase, Cochrane Database of Systematic Reviews, Web of Science and Scopus databases for systematic reviews and meta-analyses on PFMT for urinary incontinence after radical prostatectomy from inception to 3 October 2022. REVIEW METHODS: Two authors independently extracted key data from the included studies. The methodological quality of the included studies was assessed using the A Measure Tool to Assess Systematic Reviews-2 checklist. Grading of Recommendations Assessment Development and Evaluation was used to evaluate the quality of the outcomes. RESULTS: A total of 18 studies with 29,925 patients were included, all of which were of critically low methodological quality. Biofeedback therapy seemed to show additional benefits compared to PFMT alone; however, the adjunctive role of electrical stimulation remained more controversial due to the lack of strong evidence. Preoperative PFMT sometimes, but not always, showed the potential to improve urinary incontinence. PFMT with the guidance of a therapist could bring some benefits to the patient and was more acceptable to the patient, but consumed some medical resources. CONCLUSIONS: PFMT has a good effect on improving post-radical prostatectomy incontinence in men, and biofeedback can have an additional beneficial effect on patients, especially in the short-term and medium-term. However, there is insufficient evidence to suggest that electrical stimulation is beneficial for patients with urinary incontinence.

Effects of Web-Based Mindfulness-Based Interventions on Anxiety, Depression, and Stress Among Frontline Health Care Workers During the COVID-19 Pandemic: Systematic Review and Meta-Analysis.

BACKGROUND: Since 2019, the COVID-19 outbreak has spread around the world, and health care workers, as frontline workers, have faced tremendous psychological stress. OBJECTIVE: The purpose of this study is to explore whether web-based mindfulness-based interventions continue to have a positive impact on anxiety, depression, and stress among health care workers during the COVID-19 pandemic. METHODS: The inclusion criteria were as follows: (1) participants were frontline health care workers during the COVID-19 pandemic; (2) the experimental group was a web-based mindfulness-based intervention; (3) the control group used either general psychological intervention or no intervention; (4) outcome indicators included scales to assess anxiety, depression, and stress; and (5) the study type was a randomized controlled study. Studies that did not meet the above requirements were excluded. We searched 9 databases, including Web of Science, Embase, PubMed, Cochrane Library, Scopus, ScienceDirect, SinoMed, China National Knowledge Infrastructure (CNKI), and Wanfang Database, for randomized controlled studies on the effects of web-based mindfulness-based interventions on common mental disorder symptoms among health care workers from January 1, 2020, to October 20, 2022. The methodological quality of the included studies was assessed using the Physiotherapy Evidence Database scale. The Cochrane risk of bias tool was used to assess the risk of bias. Subgroup analysis was used to look for sources of heterogeneity and to explore whether the results were the same for subgroups under different conditions. Sensitivity analysis was used to verify the stability of the pooled results. RESULTS: A total of 10 randomized controlled studies with 1311 participants were included. The results showed that web-based mindfulness-based interventions were effective in reducing the symptoms of anxiety (standard mean difference [SMD]=-0.63, 95% CI -0.96 to -0.31, P<.001, I2=87%), depression (SMD=-0.52, 95% CI -0.77 to -0.26, P<.001, I2=75%), and stress (SMD=-0.20, 95% CI -0.35 to -0.05, P=.01, I2=58%) among health care workers during the COVID-19 pandemic, but with wide CIs and high heterogeneity. CONCLUSIONS: Web-based mindfulness-based interventions may be effective in reducing the symptoms of anxiety, depression, and stress among frontline health care workers during the COVID-19 pandemic. However, this effect is relatively mild and needs to be further explored by better studies in the future. TRIAL REGISTRATION: PROSPERO CRD42022343727; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=343727.

Enterolactone and trabectedin suppress epithelial ovarian cancer synergistically via upregulating THBS1.

Epithelial ovarian cancer (EOC) is the most common and fatal subtype of ovarian malignancies, with no effective therapeutics available. Our previous studies have demonstrated extraordinary suppressive efficacy of enterolactone (ENL) on EOC. A chemotherapeutic agent, trabectedin (Trabe), is shown to be effective on ovarian cancer, especially when combined with other therapeutics, such as pegylated liposomal doxorubicin or oxaliplatin. Thrombospondin 1 (THBS1), a kind of matrix glycoprotein, plays important roles against cancer development through inhibiting angiogenesis but whether it is involved in the suppression of EOC by ENL or Trabe remains unknown. To test combined suppressive effects of ENL and Trabe on EOC and possible involvement of THBS1 in the anticancer activities of ENL and Trabe. The EOC cell line ES-2 was transfected with overexpressed THBS1 by lentivirus vector. We employed tube formation assay to evaluate the anti-angiogenesis activity of ENL and of its combined use with Trabe after THBS1 overexpression and established drug intervention and xenograft nude mouse cancer models to assess the in vivo effects of the hypothesized synergistic suppression between the agents and the involvement of THBS1. Mouse fecal samples were collected for 16S rDNA sequencing and microbiota analysis. We detected strong inhibitory activities of ENL and Trabe against the proliferation and migration of cancer cells and observed synergistic effects between ENL and Trabe in suppressing EOC. ENL and Trabe, given either separately or in combination, could suppress the tube formation capability of human microvascular endothelial cells, and this inhibitory effect became even stronger with THBS1 overexpression. In the ENL plus Trabe combination group, the expression of tissue inhibitor of metalloproteinases 3 and cluster of differentiation 36 was both upregulated, whereas matrix metalloproteinase 9, vascular endothelial growth factor, and cluster of differentiation 47 were all decreased. With the overexpression of THBS1, the results became even more pronounced. In animal experiments, combined use of ENL and Trabe showed superior inhibitory effects to either single agent and significantly suppressed tumor growth, and the overexpression of THBS1 further enhanced the anti-cancer activities of the drug combination group. ENL and Trabe synergistically suppress EOC and THBS1 could remarkably facilitate the synergistic anticancer effects of ENL and Trabe.

[Analysis of variant of GLI3 gene in a child featuring autosomal dominant Pallister-Hall syndrome].

OBJECTIVE: To explore the clinical and genetic characteristics of a child with Pallister-Hall syndrome (PHS). METHODS: DNA was extracted from peripheral blood sample from the child and subjected to whole exome sequencing. Suspected variants were verified by Sanger sequencing of his family members. RESULTS: Genetic testing revealed that the child has harbored a heterozygous c.3320_3330delGGTACGAGCAG (p.G1107Afs×18) variant of the GLI3 gene. Neither parent was found to carry the same variant. CONCLUSION: The c.3320_3330delGGTACGAGCAG (p.G1107Afs×18) frameshift variant of the GLI3 gene probably underlay the pathogenesis of PHS in this child. Genetic testing should be considered for patients featuring hypothalamic hamartoma and central polydactyly.

Effects of a vegetarian diet combined with aerobic exercise on glycemic control, insulin resistance, and body composition: a systematic review and meta-analysis.

BACKGROUND: Vegetarian diets and aerobic exercise are increasingly accepted as a common way to improve lifestyle. Several studies have shown that vegetarian diets combined with aerobic exercise interventions have a significant effect on preventing and reducing the risk of metabolic diseases. METHODS: A search of the PubMed, EBSCO, Embase, CENTRAL, and Web of Science databases was conducted for comparative studies of pre- and post-vegetarian diet adoption combined with aerobic exercise interventions on glycemic control and body composition. Qualitative reviews and meta-analyses of fixed and random effects were conducted to pool available data. The results were validated by sensitivity analysis. RESULTS: A total of 27 studies were selected for meta-analysis. Combining the studies included in the meta-analysis showed a mean difference for homeostasis model assessment of insulin resistance of - 0.75 (- 1.08 to - 0.42), fasting plasma glucose of - 0.27(- 0.30 to - 0.23), waist circumference of - 1.10 (- 5.06 to 2.86) and body mass index of - 0.70 (- 1.38 to - 0.01). CONCLUSION: In summary, our findings suggest that participants who adopted a vegetarian diet combined with aerobic exercise intervention had significantly lower fasting plasma glucose and insulin levels and improved body composition compared to preintervention participants. LEVEL OF EVIDENCE: Level I, systematic review and meta-analysis.

Cu-Catalyzed Oxidative [3 + 2] Annulation of 2-(Pyridine-2-yl)acetates with Maleimides: Synthesis of 1H-Pyrrolo[3,4-b]indolizine-1,3-diones.

A novel protocol for the construction of highly functionalized indolizine derivatives, that is, 1H-pyrrolo[3,4-b]indolizine-1,3-diones (PIZDOs, 3) from 2-(pyridine-2-yl)acetates and maleimides via a regioselective oxidative [3 + 2] annulation was developed. The cascade oxidative reaction was enabled by heating a mixture of the two substrates in the presence of Ag2CO3 as an oxidant and Cu(OAc)·H2O as a catalyst in chlorobenzene. Consequently, a series of PIZDOs 3 were synthesized with high regioselectivity in moderate yields. This protocol can be used in the synthesis of functionalized PIZDOs via the one-pot oxidative annulation reaction rather than through multistep reactions, which is suitable for both combinatorial and parallel syntheses of PIZDOs.

Health-related quality of life and associated factors in Chinese menstrual migraine patients: a cross-sectional study.

BACKGROUND: Menstrual migraine is a particular form of migraine with a significant impact on the quality of life for women afflicted. Presently, no study has reported the quality of life in menstrual migraine patients. This work aims to assess the health-related quality of life and identify its associated factors among Chinese menstrual migraine patients. METHODS: The cross-sectional study group consisted of 109 patients with menstrual migraine, and the control group consisted of 397 female patients with non-menstrual migraine. In total, 506 patients completed questionnaires for demographic and clinical information, the Self-rating Idea of Suicide Scale, the Hamilton Depression Scale, the Hamilton Anxiety Scale, the Headache Impact Test-6, the Perceived Social Support Scale, the Pittsburgh Sleep Quality Index. Health-related quality of life was measured using the 36-Item Short Form Survey. RESULTS: Compared with non-menstrual migraine patients, five dimensions of health-related quality of life were all found to be significantly impaired in menstrual migraine patients. Headache frequency (ß = - 0.218, P = 0.014), the impact of headache on daily life (ß = - 0.270, P = 0.002), depression symptoms (ß = - 0.345, P < 0.001) were significantly associated with physical component summary, depression symptoms (ß = - 0.379, P < 0.001), social support (ß = 0.270, P < 0.001), suicidal ideation (ß = - 0.344, P < 0.001) were closely related to mental component summary. CONCLUSION: Menstrual migraine patients had a significantly poorer health-related quality of life in many domains than non-menstrual migraine patients. Headache frequency, the impact of headache on daily life, depression symptoms, social support, and suicidal ideation were significantly associated with health-related quality of life in menstrual migraine patients. TRIAL REGISTRATION: ChiCTR1800014343. This study was registered prospectively on 7 January 2018 at Chinese Clinical Trial registry. http://www.chictr.org.cn/showproj.aspx?proj=24526.

Monocyte-to-high-density lipoprotein-cholesterol ratio (MHR) and the risk of all-cause and cardiovascular mortality: a nationwide cohort study in the United States.

BACKGROUND: Elevated monocyte-to-high-density lipoprotein-cholesterol ratio (MHR) is relevant to higher all-cause and cardiovascular mortality in patients with coronary artery disease and other comorbidities. However, the predictive values of MHR for mortality in the general population have been underutilized. This study investigated the association of MHR with all-cause and cardiovascular mortality in the adult population of the United States. METHODS: This study included 34,335 participants (≥20 years) from the National Health and Nutrition Examination Survey 1999-2014 that were grouped according to MHR tertiles. Kaplan-Meier plots and long-rank tests were employed to investigate differences in survival among the groups. Moreover, the relationship of MHR with all-cause and cardiovascular mortality was further explored using multivariate Cox regression and restricted cubic spline analysis. RESULTS: During the average follow-up of 93.5 ± 56 months, 4310 (12.6%) participants died, with 754 (2.2%) deaths attributed to cardiovascular diseases. Kaplan-Meier analysis revealed statistically obvious differences in all-cause and cardiovascular mortality among the MHR tertiles (log-rank test: all P < 0.001). In multi-adjusted models, participants in the highest tertile of MHR had an increased risk of all-cause (hazard ratio [HR] = 1.19, 95% confidence interval [CI] 1.10-1.29) and cardiovascular mortality (HR = 1.44, 95% CI 1.17-1.77), compared to those in the lowest tertile. Furthermore, the restricted cubic spline curve indicated that MHR had a non-linear association with all-cause mortality (P < 0.001), and the inflection point of MHR was 0.006. Each 2-fold change in MHR exhibited a 32% decrease (HR = 0.68, 95%CI 0.58-0.82) and a 20% increase (HR = 1.20, 95%CI 1.13-1.27) in the risk of all-cause mortality on the left and right flanks of the inflection point, respectively. Additionally, the risk of cardiovascular mortality increased by 21% per 2-fold change in MHR (HR = 1.21, 95%CI 1.07-1.36) in a linear manner. CONCLUSIONS: MHR was significantly related to all-cause and cardiovascular mortality in the general population independent of established risk factors.

Combined 4-1BB and ICOS co-stimulation improves anti-tumor efficacy and persistence of dual anti-CD19/CD20 chimeric antigen receptor T cells.

Chimeric antigen receptor (CAR) T-cell therapy is a promising therapeutic strategy against lymphoma. However, post-treatment relapses due to antigen loss remain a challenge. Here the authors designed a novel bicistronic CAR construct and tested its functions in vitro and in vivo. The CAR construct consisted of individual anti-CD19 and anti-CD20 single-chain fragment variables equipped with ICOS-CD3ζ and 4-1BB-CD3ζ intracellular domains, respectively. The CD19 and CD20 bicistronic CAR T cells exhibited tumor lytic capacities equivalent to corresponding monospecific CAR T cells. Moreover, when stimulated with CD19 and CD20 simultaneously, the bicistronic CAR T cells showed prolonged persistence and enhanced cytokine generation compared with single stimulations. Interestingly, the authors found that the 4-1BB signal was predominant in the signaling profiles of ICOS and 4-1BB doubly activated CAR T cells. In vivo study using a CD19/CD20 double-positive tumor model revealed that the bicistronic CAR T cells were more efficient than monospecific CD19 CAR T cells in eradicating tumors and prolonging mouse survival. The authors' novel bicistronic CD19/CD20 CAR T cells demonstrate improved anti-tumor efficacy in response to dual antigen stimulations. These data provide optimism that this novel bicistronic CAR construct can improve treatment outcomes in patients with relapsed/refractory B cell malignancy.

Efficient improvement of surface displayed lipase from Rhizomucor miehei in PichiaPink™ protease-deficient system.

Lipase from Rhizomucor miehei (RML) is a promising biocatalyst used in food industry, fine chemicals, and biodiesel production. Yeast surface display allows direct application of lipase in form of whole-cell biocatalyst, avoiding purification and immobilization process, but the protease of the host cell may affect the activity of displayed lipase. Herein, we used the protease-deficient Pichia pastoris, PichiaPink™ as host to display RML efficiently. RML gene, GCW21 gene and α-factor gene were co-cloned into plasmid pPink LC/HC and transformed into protease-deficient P. pastoris. After inducution expression for 96 h, the lipase activity of displayed RML reached 121.72 U/g in proteinase-A-deficient P. pastoris harboring high-copy plasmid, which exhibited 46.7% higher than recombinant P. pastoris without protease defect. Displayed RML occurred the maximum activity at pH 8.0 and 45 °C and the optimal substrate was p-nitrophenyl octanoate. Metal ions Li+, Na+, K+, and Mg2+ of 1-10 mM had activation towards displayed RML. Displayed RML was effectively improved in PichiaPink™ protease-deficient system, which may promote the further research and development for the industrial application of RML.

Gradient Waveguide Thickness Guided-Mode Resonance Biosensor.

Portable systems for detecting biomolecules have attracted considerable attention, owing to the demand for point-of-care testing applications. This has led to the development of lab-on-a-chip (LOC) devices. However, most LOCs are developed with a focus on automation and preprocessing of samples; fluorescence measurement, which requires additional off-chip detection instruments, remains the main detection method in conventional assays. By incorporating optical biosensors into LOCs, the biosensing system can be simplified and miniaturized. However, many optical sensors require an additional coupling device, such as a grating or prism, which complicates the optical path design of the system. In this study, we propose a new type of biosensor based on gradient waveguide thickness guided-mode resonance (GWT-GMR), which allows for the conversion of spectral information into spatial information such that the output signal can be recorded on a charge-coupled device for further analysis without any additional dispersive elements. A two-channel microfluidic chip with embedded GWT-GMRs was developed to detect two model assays in a buffer solution: albumin and creatinine. The results indicated that the limit of detection for albumin was 2.92 µg/mL for the concentration range of 0.8-500 µg/mL investigated in this study, and that for creatinine it was 12.05 µg/mL for the concentration range of 1-10,000 µg/mL. These results indicated that the proposed GWT-GMR sensor is suitable for use in clinical applications. Owing to its simple readout and optical path design, the GWT-GMR is considered ideal for integration with smartphones or as miniaturized displays in handheld devices, which could prove beneficial for future point-of-care applications.

Metabolite Analysis of Jerusalem Artichoke (Helianthus tuberosus L.) Seedlings in Response to Polyethylene Glycol-Simulated Drought Stress.

Jerusalem artichokes are a perennial crop with high drought tolerance and high value as a raw material to produce biofuels, functional feed, and food. However, there are few comprehensive metabolomic studies on Jerusalem artichokes under drought conditions. METHODS: Ultra-performance liquid chromatography and tandem mass spectrometry were used to identify differential metabolites in Jerusalem artichoke seedling leaves under polyethylene glycol (PEG) 6000-simulated drought stress at 0, 18, 24, and 36 h. RESULTS: A total of 661 metabolites and 236 differential metabolites were identified at 0 vs. 18, 18 vs. 24, and 24 vs. 36 h. 146 differential metabolites and 56 common were identified and at 0 vs. 18, 24, and 36 h. Kyoto Encyclopedia of Genes and Genomes enrichment identified 236 differential metabolites involved in the biosynthesis of secondary metabolites and amino acids. Metabolites involved in glycolysis, phenolic metabolism, tricarboxylic cycle, glutamate-mediated proline biosynthesis, urea cycle, amino acid metabolism, unsaturated fatty acid biosynthesis, and the met salvage pathway responded to drought stress. CONCLUSION: A metabolic network in the leaves of Jerusalem artichokes under drought stress is proposed. These results will improve understanding of the metabolite response to drought stress in Jerusalem artichokes and develop a foundation for breeding drought-resistant varieties.

Multiple cellular responses guarantee yeast survival in presence of the cell membrane/wall interfering agent sodium dodecyl sulfate.

Sodium dodecyl sulfate (SDS), a representative anionic surfactant, is a commonly used reagent in studies of the cell membrane and cell wall. However, the mechanisms through which SDS affects cellular functions have not yet been fully examined. Thus, to gain further insights into the cellular functions and responses to SDS, we tested a haploid library of Saccharomyces cerevisiae single-gene deletion mutants to identify genes required for tolerance to SDS. After two rounds of screening, we found 730 sensitive and 77 resistant mutants. Among the sensitive mutants, mitochondrial gene expression; the mitogen-activated protein kinase signaling pathway; the metabolic pathways involved in glycoprotein, lipid, purine metabolic process, oxidative phosphorylation, cellular amino acid biosynthesis and pentose phosphate pathway were found to be enriched. Additionally, we identified a set of transcription factors related to SDS responses. Among the resistant mutants, disruption of ribosome biogenesis and translation alleviated SDS-induced cytotoxicity. Collectively, our results provided new insights into the mechanisms through which SDS regulates the cell membrane or cell wall.

Dietary yeast culture modulates immune response related to TLR2-MyD88-NF-kß signaling pathway, antioxidant capability and disease resistance against Aeromonas hydrophila for Ussuri catfish (Pseudobagrus ussuriensis).

The aim of the present study was to investigate effects of dietary yeast culture on immune response related to TLR2-MyD88-NF-kß signaling pathway, antioxidant capability and disease resistance against Aeromonas hydrophila for Ussuri catfish (Pseudobagrus ussuriensis). A total of 240 Ussuri catfish (mean weight of 7.39 ±â€¯0.32 g) were randomly distributed into four groups that fed diets containing 0 (Y0), 10 (Y1), 20 (Y2) and 30 (Y3) g kg-1 yeast culture for 8 weeks. The results indicated that dietary 10 g kg-1 yeast culture supplementation significantly down-regulated mRNA levels of TLR2, MyD88, NF-kß p65, IL-1ß and IL-8 in the liver tissue compared with the control group (P < 0.05). Simultaneously, serum lysozyme (LZM) activity, respiratory burst activity (RBA) of phagocytes, plasma alkaline phosphatase (AKP) activity and immunoglobulin M (IgM) content were significantly improved in fish fed Y1 diet (P < 0.05). Fish fed Y1 diet had significantly higher serum alternative complement pathway activity (ACH50) and plasma complement 3 (C3) content than the Y3 group (P < 0.05). However, no significant differences were observed in plasma acid phosphatase (ACP) activity and complement 4 (C4) content among the groups (P > 0.05). Fish cumulative mortality rate (CMR) in the Y1 and Y2 groups were significantly lower than that in Y0 and Y3 groups (P < 0.05), and the lowest CMR was observed in the Y1 group after challenge by A. hydrophila. The highest hepatic superoxide dismutase and glutathione peroxidase activities, total antioxidant capacity and the lowest malondialdehyde content were found in Y1 group, but no significant difference was found in hepatic catalase activity among the groups (P > 0.05). These results demonstrate that dietary 10 g kg-1 yeast culture could effectively improve the immunity, antioxidant capability and disease resistance against A. hydrophila for Ussuri catfish and could down-regulate the mRNA expression levels of pro-inflammatory cytokines modulated by TLR2-MyD88-NF-kß signaling pathway.

Neuroprotective Effect of Modified Electroconvulsive Therapy for Schizophrenia: A Proton Magnetic Resonance Spectroscopy Study.

The underlying mechanism of modified electroconvulsive therapy (MECT) treatment for drug-resistant and catatonic schizophrenia remains unclear. Here, we aim to investigate whether MECT exerts its antipsychotic effects through elevating N-acetylaspartate (NAA) concentration measured by proton magnetic resonance spectroscopy (H-MRS). Multiple-voxel H-MRS was acquired in the bilateral prefrontal cortex (PFC) and thalamus to obtain measures of neurochemistry in 32 MECT, 34 atypical antipsychotic-treated schizophrenic patients, and 34 healthy controls. We found that both MECT and atypical antipsychotic treatments showed significant antipsychotic efficacy. MECT and atypical antipsychotic treatments reversed the reduced NAA/creatine ratio (NAA/Cr) in the left PFC and left thalamus in schizophrenic patients compared with healthy controls. Furthermore, the NAA/Cr ratio after treatments was significant higher in the MECT group, but not in the medication group. Our findings demonstrate that eight times of MECT elevated the relative NAA concentration to display neuroprotective effect, which may be the underlying mechanism of rapid antipsychotic efficacy.

[Quality control of human chorionic gonadotrophin subunit dissociation].

Human chorionic gonadotrophin (hCG), a glycohormone widely used in treatment of infertility, is a heterodimer composed of an alpha- and a beta-subunit. The heterodimer could be dissociated during production and storage with an impact on its bioactivity. A CE-SDS method for quantitative analysis of hCG subunit dissociation was established in this study by optimization of a variety of method conditions including sample preparation buffer compositions, incubation temperature, separation voltage, and capillary temperature. This method was validated for good sensitivity, linearity, precision, and accuracy for both α- and ß-subunit. CE-SDS also showed much better precision and accuracy than SDS-PAGE. The method was successfully used in both recombinant hCG (r-hCG) produced by cell culture and hCG (u-hCG) derived from urine. The CE-SDS method was used in the study of hCG development and stability. Therefore, it is an useful tool for the quality control of hCG.

Investigation of the correlation between charge and glycosylation of IgG1 variants by liquid chromatography-mass spectrometry.

A recombinant IgG1 monoclonal antibody (mAb) showed multiple charge variants in a cation exchange chromatography profile. To better understand the correlation between charge heterogeneity and glycosylation, a rapid reversed phase ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method with integrated mass analysis has been developed for simultaneous determination of N-terminal pyroglutamate, C-terminal lysine truncation, and Fc glycosylation. The results show that various degrees and/or types of N-terminal pyroglutamate formation and C-terminal lysine (Lys) cleavage account for the majority of charge heterogeneity; and the charge variants showed Fc glycosylation patterns in relation to their terminal modifications. The amount of G1F decreased in the basic variants, whereas Man5 and G0F-GN increased. The complement-dependent cytotoxicity (CDC) activity of purified charge variants also suggested the potential impact of the charge differences on the glycosylation profile.