Inhibiting sEH suppresses NF-κB p65 signaling and reduces CXCL10 expression as a potential therapeutic target in HT.

BACKGROUND: Although Hashimoto's thyroiditis (HT) is one of most common autoimmune thyroid diseases, its treatment remains focused on symptom relief. The soluble epoxide hydrolase (sEH) shows potential functions as drug target in alleviating some autoimmune diseases, however, we seldom know its role in HT. METHODS: The protein expression of sEH and related downstream molecules were evaluated by immunohistochemistry, western blotting, enzyme linked immunosorbent assay or immunofluorescence staining. RNA sequencing of tissue samples was performed to analyze differential genes and dysregulated pathways in HT and controls. The thyroid follicular epithelial cells (TFECs) and rat HT model were used to verify the biological function of sEH and the inhibition role of adamantyl-ureido-dodecanoic acid (AUDA) in HT. RESULTS: The sEH was significantly up-regulated in HT patients compared with healthy individuals. Transcriptome sequencing showed cytokine-related pathways and chemokine expression, especially chemokine CXCL10 and its receptor CXCR3 were aberrant in HT patients. In TFECs and rat HT model, blocking sEH by AUDA inhibitor could effectively inhibit the autoantibody, pro-inflammatory NF-κB signaling, chemokine CXCL10/CXCR3 expression and type-1 helper CD4+ T cells. CONCLUSIONS: Our findings suggest that sEH/NF-κB p65/CXCL10-CXCR3 might be promising therapeutic targets for HT.

MAP4K4 and WT1 mediate SOX6-induced cellular senescence by synergistically activating the ATF2-TGFß2-Smad2/3 signaling pathway in cervical cancer.

SRY-box transcription factor 6 (SOX6) is a member of the SOX gene family and inhibits the proliferation of cervical cancer cells by inducing cell cycle arrest. However, the final cell fate and significance of these cell-cycle-arrested cervical cancer cells induced by SOX6 remains unclear. Here, we report that SOX6 inhibits the proliferation of cervical cancer cells by inducing cellular senescence, which is mainly mediated by promoting transforming growth factor beta 2 (TGFB2) gene expression and subsequently activating the TGFß2-Smad2/3-p53-p21WAF1/CIP1-Rb pathway. SOX6 promotes TGFB2 gene expression through the MAP4K4-MAPK (JNK/ERK/p38)-ATF2 and WT1-ATF2 pathways, which is dependent on its high-mobility group (HMG) domain. In addition, the SOX6-induced senescent cervical cancer cells are resistant to cisplatin treatment. ABT-263 (navitoclax) and ABT-199 (venetoclax), two classic senolytics, can specifically eliminate the SOX6-induced senescent cervical cancer cells, and thus significantly improve the chemosensitivity of cisplatin-resistant cervical cancer cells. This study uncovers that the MAP4K4/WT1-ATF2-TGFß2 axis mediates SOX6-induced cellular senescence, which is a promising therapeutic target in improving the chemosensitivity of cervical cancer.

Exploring the Regulatory Effect of Tegillarca granosa Polysaccharide on High-Fat Diet-Induced Non-Alcoholic Fatty Liver Disease in Mice Based on Intestinal Flora.

To explore the potential mechanism of action of Tegillarca granosa polysaccharide (TGP) in treating nonalcoholic fatty liver disease (NAFLD), the study conducts in vivo experiments using male C57BL/6 mice fed a high-fat diet while administering TGP for 16 weeks. The study measures body weight, liver weight, serum biochemical markers, pathological histology, liver lipid accumulation, oxidative stress and inflammation-related factors, lipid synthesis and metabolism-related gene and protein expression, and the composition and abundance of intestinal flora. Additionally, short-chain fatty acid (SCFAs) content and the correlation between intestinal flora and environmental factors are measured. The results show that TGP effectively reduces excessive hepatic lipid accumulation, dyslipidemia, abnormal liver function, and steatosis in the mice with NAFLD. Moreover, TGP effectively regulates intestinal flora disorder, increases the diversity of intestinal flora, and affects the relative abundance of specific bacteria while also increasing the content of SCFAs. These findings provide a basis for exploring the regulatory effect of T. granosa polysaccharide on NAFLD based on intestinal flora and highlight its potential as a natural liver nutraceutical.

De Ritis Ratio: a Potent Marker in Cancer.

BACKGROUND: The role of conventional liver function clinical laboratory marker De Ritis Ratio in evaluating the prognosis, assisting diagnosis, and monitoring therapeutic efficacy of cancer is gaining increasing attention, especially in the last decade. According to the most recent articles, the De Ritis Ratio functions have progressed, which indicates that the De Ritis Ratio appears to be a promising tumor marker. The aim of this review was to evaluate the clinical importance from studies made on this subject. METHODS: Using the search words "De Ritis Ratio", "aspartate transaminase/alanine transaminase", "aspartate transaminase", "alanine transaminase", "cancer", "prognostic significance", "diagnostic significance", and "predictive significance", a search was carried out on PubMed. Exclusion criteria were articles never published in English and articles evaluating tumor markers in cancer not involving the De Ritis Ratio. RESULTS: As a predictor of prognosis, the De Ritis Ratio is strongly associated with prognostic risk factors and can be used to assess therapeutic efficacy. As a predictor of incidence, the De Ritis Ratio could promote the prediction of the disease progression. As a biomarker, the De Ritis Ratio is more likely to improve diagnostics by being combined with other biomarkers. Therefore, since it is easily accessible, involves no additional laborious efforts, and is a relatively inexpensive marker, the De Ritis Ratio is emerging as an attractive and clinically valuable marker in cancer. CONCLUSIONS: In the review, we explore the possible mechanisms of the De Ritis Ratio related to cancer and summarize the clinical importance of the De Ritis Ratio as a promising marker for cancer.

Research on Cold Roll Forming Process of Strips for Truss Rods for Space Construction.

In this paper, a new technology for on-orbit cold forming of space truss rods is proposed. For the cold roll forming process of asymmetric cross sections of thin strips, the effects of roll gap and roll spacing on the forming of asymmetric cross sections of strips were investigated using ABAQUS simulation + experiments. The study shows the following. When forming a strip with a specific asymmetric cross section, the stresses are mainly concentrated in corners 2/4/6, with the largest strain value in corner 2. With increasing forming passes, when the roll gap is 0.3 mm, the maximum equivalent strain values are 0.09, 0.24, 0.64 sequentially. Roll gaps of 0.4 mm and 0.5 mm equivalent strain change amplitude are relatively similar, and their maximum equivalent strain values are approximately 0.07,0.15, 0.44. From the analysis of the stress-strain history of the characteristic nodes in corners 2/4/6, it can be seen that the stress and strain changes in the deformation process mainly occur at the moment of interaction between the upper and lower rollers, where the stress type of node 55786 shows two tensile types and one compressive type, the stress type of nodes 48594 and 15928 shows two compressive and one tensile type, and the strain of the three nodes is in accordance with the characteristics of plane strain. When the roll gap is about 0.4 mm, the forming of the strip is relatively good. With increased roll spacing, the strip in the longitudinal stress peak through the rollers shows a small incremental trend, but the peak stresses are 380 Mpa or so. When the roll spacing is 120 mm, the longitudinal strain fluctuation of the strip is the most serious, followed by the roll spacing at 100 mm, and the minimum at 140 mm. Combined with the fluctuation in strip edges under different roll spacings, manufacturing cost and volume and other factors, a roll spacing of 100 mm is more reasonable. It is experimentally verified that when the roll gap is 0.4 mm and the roll spacing is 100 mm, the strip is successfully prepared in accordance with the cross-section requirements. When the rolling gap is 0.3 mm, due to stress-strain concentration, the strip is prone to edge waves in forming. The top of corner 2 of the flange triangular region is susceptible to intermittent tear defects, and the crack extension mechanism is mainly based on the cleavage fracture + ductile fracture.

C2729T mutation associated with HBV mother-to-child transmission reduces HBV production via suppressing LHBs expression.

Mother-to-child transmission (MTCT) is still the main route of hepatitis B virus (HBV) infection. However, the virological factors affecting HBV MTCT have not been fully elucidated. In this study, based on a prospective cohort of mother-infant pairs with positive maternal hepatitis B surface antigen (HBsAg), we found that the average nucleotide mutation rate of HBV preS1 promoter (SPI) region in the immunoprophylaxis success group was significantly higher than that in the immunoprophylaxis failure group. Among the nucleotide mutations of the HBV SPI region, the C2729T mutation had the highest frequency. Next, we found that the C2729T mutation promoted HBsAg release but reduced HBV production by suppressing the expression of large hepatitis B surface antigen (LHBs), and overexpressing LHBs could rescue this phenomenon. Based on the fact that the C2729T mutation could alter the binding site of hepatocyte nuclear factor 1 (HNF1) in the HBV SPI region, we uncovered that such an alteration could downregulate the transcriptional activity of SPI by attenuating the binding ability of HNF1 and HBV SPI region. This study suggests that HBV C2729T mutation may contribute to the immunoprophylaxis success of HBV MTCT by reducing HBV production, which supplements the virological factors affecting HBV MTCT.

Effect of mussel polysaccharide on glucolipid metabolism and intestinal flora in type 2 diabetic mice.

BACKGROUND: Type 2 diabetes (T2D) mellitus is a major metabolic disease, and its incidence and lethality have increased significantly in recent years, making it a serious threat to human health. Among numerous previous studies, polysaccharides have been shown to alleviate the adverse effects of T2D, but there are still problems such as insufficient analysis and poor understanding of the mechanisms by which polysaccharides, especially those of marine origin, regulate T2D. METHODS: In this study, we used multiple allosteric approaches to further investigate the regulatory effects of mussel polysaccharides (MPs) on T2D and gut microbiota disorders in mice by identifying changes in genes, proteins, metabolites and target organs associated with glucolipid metabolism using an animal model of T2D fed with high-fat diets, and to explore the underlying molecular mechanisms. RESULTS: After MP intervention, serum levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and high-density lipoprotein cholesterol (HDL-C) were up-regulated, and blood glucose and lipid levels were effectively reduced in T2D mice. Activation of signaling molecules related to the upstream and downstream of the insulin PI3K/Akt signaling pathway reduced hepatic insulin resistance. The relative abundance of short-chain fatty acid (SCFA)-producing bacteria (including Akkermansia, Siraeum Eubacterium and Allobaculum) increased and harmful desulfurizing Vibrio decreased. In addition, the levels of SCFAs were increased. CONCLUSION: These results suggest that MP can increase SCFA levels by altering the abundance of intestinal flora, thereby activating the PI3K/Akt signaling pathway and exerting hypoglycemic effects. © 2023 Society of Chemical Industry.

Targeted metabolomic profiles of serum amino acids and acylcarnitines related to gastric cancer.

Background: Early diagnosis and treatment are imperative for improving survival in gastric cancer (GC). This work aimed to assess the ability of human serum amino acid and acylcarnitine profiles in distinguishing GC cases from atrophic gastritis (AG) and control superficial gastritis (SG) patients. Methods: Sixty-nine GC, seventy-four AG and seventy-two SG control patients treated from May 2018 to May 2019 in Gansu Provincial Hospitalwere included. The levels of 42 serum metabolites in the GC, AG and SG groups were detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Then, orthogonal partial least squares discriminant analysis (OPLS-DA) and the Kruskal-Wallis H test were used to identify a metabolomic signature among the three groups. Metabolites with highest significance were examined for further validation. Receiver operating characteristic (ROC) curve analysis was carried out for evaluating diagnostic utility. Results: The metabolomic analysis found adipylcarnitine (C6DC), 3-hydroxy-hexadecanoylcarnitine (C16OH), hexanoylcarnitine (C6), free carnitine (C0) and arginine (ARG) were differentially expressed (all VIP >1) and could distinguish GC patients from AG and SG cases. In comparison with the AG and SG groups, GC cases had significantly higher C6DC, C16OH, C6, C0 and ARG amounts. Jointly quantitating these five metabolites had specificity and sensitivity in GC diagnosis of 98.55% and 99.32%, respectively, with an area under the ROC curve (AUC) of 0.9977. Conclusion: This study indicates C6DC, C16OH, C6, C0 and ARG could effectively differentiate GC cases from AG and SG patients, and may jointly serve as a valuable circulating multi-marker panel for GC detection.

Urinary Biomarkers for the Noninvasive Detection of Gastric Cancer.

Gastric cancer (GC) is associated with high morbidity and mortality rates. Thus, early diagnosis is important to improve disease prognosis. Endoscopic assessment represents the most reliable imaging method for GC diagnosis; however, it is semi-invasive and costly and heavily depends on the skills of the endoscopist, which limit its clinical applicability. Therefore, the search for new sensitive biomarkers for the early detection of GC using noninvasive sampling collection methods has attracted much attention among scientists. Urine is considered an ideal biofluid, as it is readily accessible, less complex, and relatively stable than plasma and serum. Over the years, substantial progress has been made in screening for potential urinary biomarkers for GC. This review explores the possible applications and limitations of urinary biomarkers in GC detection and diagnosis.

Establishment of Reference Values for Pepsinogens in Healthy Subjects of the Gansu Province: a Cross-Sectional Study.

BACKGROUND: At present, the pepsinogen reference values applicable to subjects from Gansu province have not been established. Therefore, the current study aimed to establish reference values for PGI, PGII, and the PGI/ PGII ratio in Gansu Province, Northwest China. METHODS: The present study was a cross-sectional study. Following screening in the physical examination center of Gansu Provincial Hospital, 2,130 healthy subjects were enrolled (age range 18 - 88 years; BMI range 15.35 - 38.89 kg/m2) from March 2018 to December 2020. Serum PGI and PG II concentration levels were detected by chemiluminescence. The reference values were defined according to age and gender by non-parametric 95th percentile intervals. RESULTS: The increase in age caused a gradual increase in the levels of PG I and PG II, while PG I/PG II ratio gradually decreased. The PG I, PG II and PG I/PG II ratio in males were significantly higher than those in females. The reference values for PG I, PG II and PG I/PG II ratio in males: < 40 years old were 22.79 - 119.79 ng/mL, 3.02 - 21.57 ng/mL, and 2.99 - 10.25, respectively; ≥ 40 years old were 17.58 - 125.12 ng/mL, 3.70 - 25.84 ng/mL, and 1.52 - 10.53, respectively. The reference values for PG I, PG II, and PG I/PG II ratio in females: < 40 years old were 22.57 - 103.90 ng/mL, 3.17 - 20.73 ng/mL, and 2.28 - 10.46, respectively; ≥ 40 years old were 14.24 - 117.81 ng/mL, 3.36 - 29.57 ng/mL, and 1.26 - 9.85, respectively. CONCLUSIONS: The present study determined the missing reference values of serum PGs for healthy subjects of different gender and ages in Gansu Province.

MAP4K4 mediates the SOX6-induced autophagy and reduces the chemosensitivity of cervical cancer.

There are nearly 40% of cervical cancer patients showing poor response to neoadjuvant chemotherapy that can be induced by autophagy, however, the underlying mechanism has not yet been fully clarified. We previously found that Sex-determining region of Y-related high-mobility-group box 6 (SOX6), a tumor suppressor gene or oncogene in several cancers, could induce autophagy in cervical cancer. Accordingly, this study aims to investigate the mechanism of SOX6-induced autophagy and its potential significance in the platinum-based chemotherapy of cervical cancer. Firstly, we found that SOX6 could promote autophagy in cervical cancer cells depending on its HMG domain. Mitogen-activated protein kinase kinase kinase kinase-4 (MAP4K4) gene was identified as the direct target gene of SOX6, which was transcriptionally upregulated by binding the HMG domain of SOX6 protein to its double-binding sites within MAP4K4 gene promoter. MAP4K4 mediated the SOX6-induced autophagy through inhibiting PI3K-Akt-mTOR pathway and activating MAPK/ERK pathway. Further, the sensitivity of cervical cancer cells to cisplatin chemotherapy could be reduced by the SOX6-induced autophagy in vitro and in vivo, while such a phenomenon could be turned over by autophagy-specific inhibitor and MAP4K4 inhibitor, respectively. Moreover, cisplatin itself could promote the expression of endogenous SOX6 and subsequently the MAP4K4-mediated autophagy in cervical cancer cells, which might in turn reduce the sensitivity of these cells to cisplatin treatment. These findings uncovered the underlying mechanism and potential significance of SOX6-induced autophagy, and shed new light on the usage of MAP4K4 inhibitor or autophagy-specific inhibitor for sensitizing cervical cancer cells to the platinum-based chemotherapy.

Neuropeptide S Attenuates the Alarm Pheromone-Evoked Defensive and Risk Assessment Behaviors Through Activation of Cognate Receptor-Expressing Neurons in the Posterior Medial Amygdala.

Neuropeptide S (NPS) acts by activating its cognate receptor (NPSR). High level expression of NPSR in the posterior medial amygdala suggests that NPS-NPSR system should be involved in regulation of social behaviors induced by social pheromones. The present study was undertaken to investigate the effects of central administration of NPS or with NPSR antagonist on the alarm pheromone (AP)-evoked defensive and risk assessment behaviors in mice. Furthermore, H129-H8, a novel high-brightness anterograde multiple trans-synaptic virus, c-Fos and NPSR immunostaining were employed to reveal the involved neurocircuits and targets of NPS action. The mice exposed to AP displayed an enhancement in defensive and risk assessment behaviors. NPS (0.1-1 nmol) intracerebroventricular (i.c.v.) injection significantly attenuated the AP-evoked defensive and risk assessment behaviors. NPSR antagonist [D-Val5]NPS at the dose of 40 nmol completely blocked the effect of 0.5 nmol of NPS which showed the best effective among dose range. The H129-H8-labeled neurons were observed in the bilateral posterodorsal medial amygdala (MePD) and posteroventral medial amygdala (MePV) 72 h after the virus injection into the unilateral olfactory bulb (OB), suggesting that the MePD and MePV receive olfactory information inputs from the OB. The percentage of H129-H8-labeled neurons that also express NPSR were 90.27 ± 3.56% and 91.67 ± 2.46% in the MePD and MePV, respectively. NPS (0.5 nmol, i.c.v.) remarkably increased the number of Fos immunoreactive (-ir) neurons in the MePD and MePV, and the majority of NPS-induced Fos-ir neurons also expressed NPSR. The behavior characteristic of NPS or with [D-Val5]NPS can be better replicated in MePD/MePV local injection within lower dose. The present findings demonstrated that NPS, via selective activation of the neurons bearing NPSR in the posterior medial amygdala, attenuates the AP-evoked defensive and risk assessment behaviors in mice.

The Role of Splicing Factor SF3B4 in Congenital Diseases and Tumors.

In eukaryotes, spliceosomes catalyze the splicing of pre-mRNA to mature mRNA. As the core subunit of U2 spliceosome, splicing factor SF3b4 plays not only a crucial role in the splicing process, but also a role in transcription, translation, and cell signal transduction, and participates in the regulation of cell cycle, cell differentiation, and immune deficiency. In recent years, more and more research studies on SF3b4-related diseases, such as Nager syndrome and cancer, have been conducted. It has been found that SF3b4 mutations led to abnormal cell growth and were involved in the development and occurrence of these diseases. In this review, the diseases, mainly congenital diseases and tumors, in which SF3B4 is involved and the pathogenesis of them were summarized, aiming to provide a better understanding of the roles of SF3B4 in the prevention, diagnosis, and treatment of diseases in the future.

Oxygenation properties and underlying molecular mechanisms of hemoglobins in plateau zokor (Eospalax baileyi).

The plateau zokor (Eospalax baileyi) is a species of subterranean rodent endemic to the Tibetan Plateau. It is well adapted to the cold and hypoxic and hypercapnic burrow. To study the oxygenation properties of plateau zokor hemoglobins (Hbs), we measured intrinsic Hb-O2 affinities and their sensitivities to pH (Bohr effect); CO2; Cl-, 2,3-diphosphoglycerate (DPG); and temperature using purified Hbs from zokor and mouse. The optimal deoxyHb model of plateau zokor was constructed and used to study its structural characteristics by molecular dynamics simulations. O2 binding results revealed that plateau zokor Hbs exhibit remarkably high intrinsic Hb-O2 affinity, low CO2 effects compared with human and the relatively low anion allosteric effector sensitivities (DPG and Cl-) at normal temperature, which would safeguard the pulmonary Hb-O2 loading under hypoxic and hypercapnic conditions. Furthermore, the high anion allosteric effector sensitivities at low temperature and low temperature sensitivities of plateau zokor Hbs would facilitate the releasing of O2 in cold extremities and metabolic tissues. However, the high Hb-O2 affinity of plateau zokor is not compensated by high pH sensitivity as the Bohr factors of plateau zokor Hbs were as low as those of mouse. The results of molecular dynamics simulations revealed the reduced hydrogen bonding between the α1ß1- and α2ß2-dimer interface of deoxyHb in zokor compared with mouse. It may be the primary mechanism of the high intrinsic Hb-O2 affinities in zokor. Specifically, substitution of the 131Ser→Asn in the α2-chain weakened the connection between α1- and ß2-subunit.

[Expression, purification and characterization of human prorelaxin-like-protein H2 in Escherichia coli].

The cDNA gene encoding human prorelaxin H2 was subcloned from plasmid pMAL p2X-hRLX H2 into the EcoRI and BamHI sites of prokaryotic expression vector pBV220, resulting in the recombinant plasmid pBV220-hPRLX H2, which was subsequently transformed into Escherichia coli DH5arg;. The target gene was highly expressed in the form of inclusion body by thermoinduction at 42 degrees. Recombinant human Met-prorelaxin-like-protein H2 was refolded in vitro, purified by Sephadex G-75 gel filtration chromatography and reverse phase fast protein liquid chromatography. The yield was approximately 2-3 mg/L. The purified recombinant human Met-prorelaxin-like-protein H2 was shown to be a single band by 15% SDS-PAGE and gave correct amino acid composition of Met-prorelaxin. Molecular weight of the purified recombinant human Met-prorelaxin-like-protein H2 was measured to be 18 390.4 (calculated theoretical value 18 392.3) by matrix assisted laser desorption ionization time-of-flight mass spectroscopy.

Acidic residues on the N-terminus of proinsulin C-Peptide are important for the folding of insulin precursor.

To investigate the role of C-peptide in the folding of insulin precursor, a series of C-peptide mutant proinsulin genes were constructed, overexpressed in Escherichia coli and the proteins purified. Correct disulfide linkages of these proteins were confirmed by both tryptic peptide mapping and insulin receptor binding analyses. In vitro refolding experiments were performed with the purified proteins and showed that mutations on the glycine-rich middle segment of C-peptide, GGGPGAG, and deletion of the C-terminal pentapeptide, EGSLQ, as well as mutations on the two pairs of dibasic residues at the two ends of C-peptide did not significantly affect the refolding yields. However, both alanine replacement mutation and deletion of three highly conserved acidic residues (EAED) at the N-terminus of the C-peptide resulted in serious aggregation during refolding. The results indicate that the highly conserved acidic N-terminal part of C-peptide is very important for insulin precursor folding, and that C-peptide may have some intramolecular chaperone-like function in the folding of insulin precursor.