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Despite the long availability of a traditional prophylactic vaccine containing the HBV surface antigen (HBsAg) and aluminum adjuvant, nearly 10% of the population remains unable to generate an effective immune response. Previous studies have indicated that hepatitis B virus (HBV) PreS2-S is abundant in T/B cell epitopes, which induces a stronger immune response than HBsAg, particularly in terms of cytotoxic T lymphocyte (CTL) reaction. In the current study, the HBV PreS2-S gene encoding an extra 26 amino acids (PreS2 C-terminus) located at the N-terminus of HBsAg was cloned into the pVCH1300 expression vector. PreS2-S expressed in the methylotrophic yeast, Hansenula polymorpha, was produced at a yield of up to 250 mg/L. Subsequent purification steps involved hydrophobic adsorption to colloidal silica, ion-exchange chromatography and density ultracentrifugation. The final product was obtained with a total yield of ∼ 15% and purity of ∼ 99%. In keeping with previous studies, ∼ 22 nm viruslike particles were detected using electron microscopy. The generated PreS2-S antigen will be further studied for efficacy and safty in animals.

Fermentation is an important strategy for food preservation, and fermented foods are the result of the joint action of a diverse range of microorganisms that impart a special flavor to the product. Quorum sensing system (QS) is a highly targeted and universal microbial regulation mechanism, which is based on population density and regulates the behavior of multiple gene expressions by sensing the concentration of autoinducers (AI). According to different types of AIs, QS systems are classified into three major categories. The article provided an overview of fermented foods and the three major types of QS systems. The positive effects of QS systems on microbial co-culture, the prevention and control of harmful microorganisms, and the effects on fermented food quality were described. From the perspective of QS, the article broadens the research ideas of microorganisms in fermented foods, and lays the foundation for further improving the application of microorganisms in food production. [ABSTRACT FROM AUTHOR]

Objective: To observe the effect of Jiawei Danggui Buxuetang on the expression of microRNA-155 (miR-155) and cytokine signal transduction inhibitor 1 (SOCS1) in renal tissue of rats with diabetic nephropathy (DKD), and to explore the mechanism of Jiawei Danggui Buxuetang in the treatment of DKD. Methods: Firty 6-week-old SPF male SD rats were randomly divided into normal group (CON) and model group. The model group was fed with high-sugar and high-fat diet for 6 weeks, and the DKD rat model was established by intraperitoneal injection of streptozotocin (STZ) (35 mg/kg). Then the rats were randomly divided into model group (MOD), low dose group (DBTL), medium dose group (DBTM),high dose group (DBTH) and irbesartan group (IRB). Each group was given intragastric administration for 20 weeks, during which CON group and MOD group were given the same amount of normal saline. Urine samples were collected after administration, and the ratio of urinary Microalbumin to urinary creatinine (UACR) was analyzed by automatic analyzer. The renal changes in each group were observed by hematoxylin-eosin (HE) staining, Masson pine (Masson) trichromatic staining and PASM staining. The expression of miR-155 and SOCS1mRNA in renal tissue of DKD rats was detected by Real-time PCR, and the expression of SOCS1 was detected by Western blotting. Results: By comparison with CON group, UACR value, MIR-155 mRNA expression and SOCS1 mRNA expression in MOD group increased significantly, while SOCS1 mRNA expression decreased significantly compared with MOD group, UACR decreased significantly in IRB group and DBT group, SOCS1 expression increased significantly in DBTM group, DBTH group and IRB group. The expression of MIR-155 decreased significantly in all treatment groups (P<0.05), and the expression of SOCS1 in DBTM group, DBTH group and IRB group increased significantly (P<0.05). Conclusion: Jiawei Danggui Buxuetang can reduce the expression of miR-155 in high glucose and increase the expression of SOCS1 in renal tissue, so as to significantly reduce renal damage and play a role in renal protection. [ABSTRACT FROM AUTHOR]

Understanding the causes of tumorigenesis and progression in triple-receptor negative breast cancer (TNBC) can help the design of novel and personalized therapies and prognostic assessments. Abnormal RNA modification is a recently discovered process in TNBC development. TNBC samples from The Cancer Genome Atlas database were categorized according to the expression level of NAT10, which drives acetylation of cytidine in RNA to N(4)-acetylcytidine (ac4C) and affects mRNA stability. A total of 703 differentially expressed long non-coding RNAs (lncRNAs) were found between high- and low-expressed NAT10 groups in TNBC. Twenty of these lncRNAs were significantly associated with prognosis. Two breast cancer tissues and their paired normal tissues were sequenced at the whole genome level using acetylated RNA immunoprecipitation sequencing (acRIP-seq) technology to identify acetylation features in TNBC, and 180 genes were significantly differentially ac4c acetylated in patients. We also analyzed the genome-wide lncRNA expression profile and constructed a co-expression network, containing 116 ac4C genes and 1080 lncRNAs. Three of these lncRNAs were prognostic risk lncRNAs affected by NAT10 and contained in the network. The corresponding reciprocal pairs were "LINC01614-COL3A1", "OIP5-AS1-USP8", and "RP5-908M14.9-TRIR". These results indicate that RNA ac4c acetylation involves lncRNAs and affects the tumor process and prognosis of TNBC. This will aid the prediction of drug targets and drug sensitivity. [ABSTRACT FROM AUTHOR]

Cisplatin is widely used as a chemotherapeutic drug to treat various solid tumors. However, it often induces severe side effects, including nephrotoxicity, which limits its application in clinical settings. Furthermore, the underlying mechanisms of action are unclear. Here, we applied whole-transcriptome RNA sequencing to a cisplatin-induced acute kidney injury (CP-AKI) mouse model to evaluate competing endogenous RNA (ceRNA) networks. We found 4460 mRNAs, 1851 long non-coding RNAs, 101 circular RNAs, and 102 microRNAs significantly differentially expressed between CP-AKI and control mice. We performed gene set enrichment analysis to reveal the biological functions of the mRNAs and constructed non-coding RNA-associated ceRNA networks in CP-AKI mice. Two ceRNA regulatory pathways, Lhx1os-203/mmu-miR-21a-3p/Slc7a13 and circular RNA_3907/mmu-miR-185-3p/Ptprn, were validated using quantitative real-time PCR. The protein–protein interaction network indicated that Il6, Cxcl1, Cxcl2, and Plk1 serve as hub genes and are highly connected with the inflammatory response or DNA damage. Transcription factors, such as Stat3, Cebpb, and Foxm1, regulate gene expression levels in CP-AKI. Our study provides insight into non-coding RNA-associated ceRNA networks and mRNAs in CP-AKI and identifies potential treatment targets. [ABSTRACT FROM AUTHOR]

• Discovery of increased Trim27 expression in asthma. • The deletion of Trim27 effectively relieved ovalbumin (OVA)-induced airway hyperresponsiveness (AHR) and lung tissue histopathological changes. • Verification of Trim27's impact on asthma symptoms and Revealing Trim27's role in promoting NLRP3 inflammasome activation. Asthma is a prevalent chronic respiratory disease, yet understanding its ecology and pathogenesis remains a challenge. Trim27, a ubiquitination ligase belonging to the TRIM (tripartite motif-containing) family, has been implicated in regulating multiple pathophysiological processes such as inflammation, oxidative stress, apoptosis, and cell proliferation. However, the role of Trim27 in asthma has not been investigated. Our study found that Trim27 expression significantly increases in the airway epithelium of asthmatic mice. Knockdown of Trim27 expression effectively relieved ovalbumin (OVA)-induced airway hyperresponsiveness (AHR) and lung tissue histopathological changes. Moreover, Trim27 knockdown exhibited a significant reduction in airway inflammation and oxidative stress in asthmatic mice, and in vitro analysis confirmed the favorable effect of Trim27 deletion on inflammation and oxidative stress in mouse airway epithelial cells. Furthermore, our study revealed that deletion of Trim27 in MLE12 cells significantly decreased NLRP3 inflammasome activation, as evidenced by reduced expression of NLRP3, ASC, and pro-IL-1β mRNA. This downregulation was reversed when Trim27, but not its mutant lacking ubiquitination ligase activity, was replenished in these cells. Consistent with these findings, protein levels of NLRP3, pro-caspase-1, pro-IL-1β, cleaved-caspase-1, and cleaved-IL-1β were higher in Trim27-replenished cells compared to cells expressing Trim27C/A. Functionally, the downregulation of IL-1β and IL-18 levels induced by Trim27 deletion was rescued by replenishing Trim27. Overall, our findings provide evidence that Trim27 contributes to airway inflammation and oxidative stress in asthmatic mice via NLRP3 inflammasome activation, providing crucial insights into potential therapeutic interventions targeting Trim27 as a way to treat asthma. [ABSTRACT FROM AUTHOR]

Quorum sensing (QS) exists in bacteria to communicate with each other and regulate group behaviors in a cell density-dependent manner, which uses signal molecule autoinducer-2 (AI-2) to intra- and inter-species communication. Effects of exogenous AI-2 on biofilm formation and environmental tolerance in Lactobacillus plantarum are the focus of this review. The responses to the exogenous AI-2 cross multiple physiological metabolic behaviors involving the bacteria growth, morphological characterization, biofilm development, extracellular polysaccharides (EPS) amount and related genes expression as well as the environmental stresses tolerance. The cell surface was smoother in the AI-2 supplemented treatments than without AI-2. Meanwhile, AI-2 had ability to promote the growth and formation of biofilm by increasing the yield of EPS, the main components of biofilm. The changes in lamC and ftsH gene expression point to altered regulation for hydrolysis process of polysaccharides as well as the potential for enhanced biofilm formation. The presence of AI-2 also significantly improved (p < 0.01) the tolerance of bile salts in L. plantarum , but the same results did not appear in acid tolerance. In conclusion, AI-2 supplementation could improve the biofilm formation and bile salts tolerance in L. plantarum , and this effect was likely modulated by facilitating EPS production and suppression polysaccharides hydrolysis. • The addition of AI-2 made the cell surface of L. plantarum smoother. • Exogenous AI-2 had positive impact on L. plantarum growth and biofilm formation. • Be promoted biofilm caused by more EPS secretion and reduced polysaccharide hydrolysis. • The ability of the strain to resist bile salts has increased under the action of AI-2. [ABSTRACT FROM AUTHOR]

[Display omitted] • S. cerevisiae YE4 influenced E. faecium and L. fermentum physiological behaviors. • S. cerevisiae YE4 caused changes in AI-2 activity and QS gene expression. • S. cerevisiae YE4 effected the proteins expression of cell wall formation and QS. The interaction mode between lactic acid bacteria (LAB) and yeast in a fermentation system directly determines the quality of the products, thus understanding their mode of interaction can improve product quality. The present study investigated the effects of Saccharomyces cerevisiae YE4 on LAB from the perspectives of physiology, quorum sensing (QS), and proteomics. The presence of S. cerevisiae YE4 slowed down the growth of Enterococcus faecium 8–3 but had no significant effect on acid production or biofilm formation. S. cerevisiae YE4 significantly reduced the activity of autoinducer-2 at 19 h in E. faecium 8–3 and at 7–13 h in Lactobacillus fermentum 2–1. Expression of the QS-related genes luxS and pfs was also inhibited at 7 h. Moreover, a total of 107 E. faecium 8–3 proteins differed significantly in coculture with S. cerevisiae YE4—these proteins are involved in metabolic pathways including biosynthesis of secondary metabolites; biosynthesis of amino acids; alanine, aspartate, and glutamate metabolism; fatty acid metabolism; and fatty acid biosynthesis. Among them, proteins involved in cell adhesion, cell wall formation, two-component systems, and ABC transporters were detected. Therefore, S. cerevisiae YE4 might affect the physiological metabolism of E. faecium 8–3 by affecting cell adhesion, cell wall formation, and cell–cell interactions. [ABSTRACT FROM AUTHOR]

Osteogenesis from preosteoblasts is important for bone tissue engineering. MicroRNAs are a class of endogenous small RNA molecules that potentially modulate osteogenesis. In this study, we found that miR-155 expression was downregulated in a time-dependent manner in cells of the preosteoblast cell line MC3T3-E1 after osteogenic induction using bone morphogenetic protein 2 (BMP2). Transfection with miR-155 decreased alkaline phosphatase (ALP) activity, ALP expression, and the staining intensity of Alizarin Red in MC3T3-E1 cells treated with BMP2, whereas treatment with miR-155 inhibitor promoted BMP2-induced osteoblast differentiation. The luciferase assay confirmed that miR-155 can bind to the 3′ untranslated region of SMAD5 mRNA. miR-155 transfection significantly decreased the expression of SMAD5 protein and mRNA in MC3T3-E1 cells under control media and the p-SMAD5 protein level during osteogenesis. After transfecting cells with the SMAD5 overexpression plasmids, the inhibitory effect of miR-155 on osteogenesis was significantly attenuated. In conclusion, miR-155 inhibited osteoblast differentiation by downregulating the translation of SMAD5 in mouse preosteoblast cells. Inhibition of miR-155 promoted osteogenic potential and thus it can be used as a potential target in the treatment of bone defects. [ABSTRACT FROM AUTHOR]

Celastrol is a triterpenoid compound extracted from the Chinese herb Tripterygium wilfordii Hook F. Previous research has revealed its anti-oxidant, anti-inflammatory, anti-cancer and immunosuppressive properties. Here, we investigated whether celastrol inhibits oxidized low-density lipoprotein (oxLDL) induced oxidative stress in RAW 264.7 cells. In addition, the effect of celastrol on atherosclerosis in vivo was assessed in apolipoprotein E knockout (apoE−/−) mouse fed a high-fat/high-cholesterol diet (HFC). We found that celastrol significantly attenuated oxLDL-induced excessive expression of lectin-like oxidized low density lipoprotein receptor-1(LOX-1) and generation of reactive oxygen species (ROS) in cultured RAW264.7 macrophages. Celastrol also decreased IκB phosphorylation and degradation and reduced production of inducible nitric oxide synthase (iNOS), nitric oxide (NO) and proinflammatory cytokines such as tumor necrosis factor (TNF)-α and IL-6. Celastrol reduced atherosclerotic plaque size in apoE−/− mice. The expression of LOX-1 within the atherosclerotic lesions and generation of superoxide in mouse aorta were also significantly reduced by celastrol while the lipid profile was not improved. In conclusion, our results show that celastrol inhibits atherosclerotic plaque developing in apoE−/− mice via inhibiting LOX-1 and oxidative stress. [ABSTRACT FROM AUTHOR]

Aim To establish an improved substrate for an indirect immunofluorescence test ( IIF) to detect anti- Sm antibody. Methods Full-length Smith protein D1( Sm- D1) complementary DNA was obtained from human larynx carcinoma cell line HEp-2 by reverse transcription - polymerase chain reaction ( RT- PCR) and cloned into the mammalian expression vector p EGFP- C1. The recombinant plasmid p EGFP- Sm- D1 was transfected into HEp-2 cells. The expression, localization and antigenicity of fusion proteins of green fluorescent protein ( GFP) in transfected cells were confirmed by means of immunoblotting ( IBT), confocal fluorescence microscopy and IIF analysis. Transfected HEp-2 cells were analyzed with reference serum and compared with untransfected HEp-2 cells by IIF. Results Stable expression of the Sm- D1- GFP was maintained for more than ten generations. This Sm- D1- GFP showed the antigenicity of Sm- D1 with a characteristic phenotype in IIF. Six of 12 serum specimens from systemic lupus erythematosus contained both 29/28 and 13.5 k Da proteins and showed characteristic immunofluorescent patterns. The same phenomenon appeared in 3/6 serum samples which contained 29/28 k Da proteins only. Sera from 10 healthy donors did not react with HEp- Sm- D1 or HEp-2 at 1:80 attenuant degrees. No alteration in expression, localization and morphology was observed when HEp- Sm- D1 or HEp-2 interacted with the reference sera which could react with Ro/ SSA, La/ SSB, β2 GP1, centromere, histone, and Scl-70 antibodies in routine IIF tests. Conclusion As a new kind of substrate of IIF, HEp- Sm- D1 can be used to detect anti- Sm antibodies. Transfected HEp-2 cells keep the immunofluorescent property of HEp-2 cells in immunofluorescence anti-nuclear antibody ( IFANA) test and could potentially be used as substrate for routine IFANA detection. [ABSTRACT FROM AUTHOR]

The suprachiasmatic nucleus (SCN) is the master circadian pacemaker that drives body temperature rhythm. Time-restricted feeding (TRF) has potential as a preventative or therapeutic approach against many diseases. The potential side effects of TRF remain unknown. Here we show that daily 4-h TRF treatment in mice can severely impair body temperature homeostasis and can result in lethality. Nearly half of the mice died at 21 °C, and all mice died at 18 °C during 4-h TRF. This effect was modulated by the circadian clock and was associated with severe hypothermia due to loss of body temperature homeostasis. Remarkably, disrupting the circadian clock by SCN lesions or constant light rescued lethality during TRF. Analysis of circadian gene expression in the dorsomedial hypothalamus (DMH) demonstrated that TRF reprograms rhythmic transcriptome in DMH and suppresses expression of genes, such as Ccr5 and Calcrl, which are involved in thermoregulation. Together, our results demonstrated a potent effect of TRF on body temperature homeostasis and the role of SCN function in this process. Our results further suggested that circadian arrhythmicity can have potential beneficial effects under certain stress conditions. [ABSTRACT FROM AUTHOR]

Pompano fishes have been widely farmed worldwide. As a representative commercial marine species of the Carangidae family, the golden pompano (Trachinotus blochii) has gained significant popularity in China and worldwide. However, because of rapid growth and high-density aquaculture, the golden pompano has become seriously threatened by various diseases. Cell lines are the most cost-effective resource for in vitro studies and are widely used for physiological and pathological research owing to their accessibility and convenience. In this study, we established a novel immortal cell line, GPF (Golden pompano fin cells). GPF has been passaged over 69 generations for 10 months. The morphology, adhesion and extension processes of GPF were evaluated using light and electron microscopy. GPF cells were passaged every 3 days with L-15 containing 20 % fetal bovine serum (FBS) at 1:3. The optimum conditions for GPF growth were 28 °C and a 20 % FBS concentration. DNA sequencing of 18S rRNA and mitochondrial 16S rRNA confirmed that GPF was derived from the golden pompano. Chromosomal analysis revealed that the number pattern of GPF was 48 chromosomes. Transfection experiments demonstrated that GPF could be utilized to express foreign genes. Furthermore, heavy metals (Cd, Cu, and Fe) exhibited dose-dependent cytotoxicity against GPF. After polyinosinic-polycytidylic acid (poly I:C) treatment, transcription of the retinoic acid-inducible gene I-like receptor (RLR) pathway genes, including mda5 , mita , tbk1 , irf3 , and irf7 increased, inducing the expression of interferon (IFN) and anti-viral proteins in GPF cells. In addition, lipopolysaccharide (LPS) stimulation up-regulated the expression of inflammation-related factors, including myd88 , irak1 , nfκb , il1β , il6 , and cxcl10 expression. To the best of our knowledge, this is the first study on the immune response signaling pathways of the golden pompano using an established fin cell line. In this study, we describe a preliminary investigation of the GPF cell line immune response to poly I:C and LPS, and provide a more rapid and efficient experimental material for research on marine fish immunology. • GPF was established and became a stable fin cell line of Trachinotus blochii. • GPF can be used to assess the cytotoxicity of heavy metals. • GPF can be utilized to express foreign genes with 32 % transfection efficiency. • GPF's immune response to LPS and poly I: C challenge proved it can be used for in vitro immunology study. [ABSTRACT FROM AUTHOR]

Abstract: MiR-1, miR-133a, and miR-206a have been identified as muscle-specific miRNAs. They play multiple crucial roles in the regulation of muscle development. Here, we show that these miRNAs were differentially expressed during the larval development of flounder, and specifically expressed in skeletal muscle and heart in adult tissues/organs. The expression levels of these miRNAs were significantly changed by thyroid hormone (TH) or thiourea (TU) treatment during metamorphosis from 17 dph (days post hatching) to 42 dph. In addition, the expression levels of MyoD and Myf5 mRNAs markedly increased at 14 dph (pre-metamorphosis) compared to metamorphic stages, and their expression levels are far above the myogenin during larval development. Moreover, these MRFs (myogenic regulatory factors) expression were directly or indirectly regulated by thyroid hormone or thiourea during metamorphosis. All the results suggest that miRNAs and MRFs might be involved in signaling pathway of TH or TU-mediated flounder metamorphosis. [Copyright &y& Elsevier]

To explore the effects of different quorum sensing inhibitors (QSIs) on the biofilm formation of Lactobacillus fermentum , a high-production signal molecule AI-2 screened by the research group, was taken as the research object and subjected to various QSIs. Three QSIs, namely 100 μmol/L d -galactose, 200 μmol/L d -ribose, and furanone, could inhibit the AI-2 activity of L. fermentum and the biofilm formation in a certain concentration range but not significantly affect the growth of the strain. Scanning electron microscopy and fluorescence microscopy revealed that each QSI did not significantly affect the bacterial cell morphology in planktonic and biofilm states at the optimal inhibitory final concentration, although they reduced the adhesion of the strain. The expression of luxS , a key gene of quorum sensing, was downregulated. When biofilm production increased, QSIs were added, and the activity of the AI-2 was consistent with the change in biofilm production. Whole-genome technology revealed that QSIs inhibited the expression of the luxS , resulting in restriction of pyruvate synthesis and downregulation of the expression of genes such as gapA , pgmB , ribBA , and ribF , which in turn affected the transport and metabolism of carbohydrates and coenzymes in the strain and ultimately reduced the biofilm formation of L. fermentum. • The signal molecule AI-2 of Lactobacillus fermentum does not depend on cell density. • The effect of quorum sensing inhibitors on L. fermentum biofilm is strain-specific. • Different QSIs reduce biofilm formation by interfering with the transduction of AI-2. • QSIs may affect the transport and metabolism of carbohydrates and coenzymes in cell. [ABSTRACT FROM AUTHOR]

It has been revealed that intersubject variability (ISV) in intrinsic functional connectivity (FC) is associated with a wide variety of cognitive and behavioral performances. However, the underlying organizational principle of ISV in FC and its related gene transcriptional profiles remain unclear. Using resting-state fMRI data from the Human Connectome Project (299 adult participants) and microarray gene expression data from the Allen Human Brain Atlas, we conducted a transcription-neuroimaging association study to investigate the spatial configurations of ISV in intrinsic FC and their associations with spatial gene transcriptional profiles. We found that the multimodal association cortices showed the greatest ISV in FC, while the unimodal cortices and subcortical areas showed the least ISV. Importantly, partial least squares regression analysis revealed that the transcriptional profiles of genes associated with human accelerated regions (HARs) could explain 31.29% of the variation in the spatial distribution of ISV in FC. The top-related genes in the transcriptional profiles were enriched for the development of the central nervous system, neurogenesis and the cellular components of synapse. Moreover, we observed that the effect of gene expression profile on the heterogeneous distribution of ISV in FC was significantly mediated by the cerebral blood flow configuration. These findings highlighted the spatial arrangement of ISV in FC and their coupling with variations in transcriptional profiles and cerebral blood flow supply. [ABSTRACT FROM AUTHOR]