Objective TXN is a thioredoxin (TXN) that participates in many redox reactions and plays a crucial role in various signaling pathways. However, the role of TXN in many cancers is still unclear. The objective of this study is to investigate and visualize the diagnostic, prognostic, and immunological implications of TXN expression across various cancer types. Methods The clinical data were downloaded from the cancer genome mapping project(TCGA) database to analyze the expression level of TXN in pan cancer, and the expression level was preliminarily verified by human protein mapping (HPA)(https://www.proteinatlas.org/)database. The ESTIMATE algorithm and CIBERSORT algorithm were applied to calculate the correlation between TXN expression and immune cell infiltration. The correlation between TXN and microsatellite instability (MSI) and tumor mutation burden (TMB) was analyzed using Spearman method. Gene Set Enrichment Analysis (GSEA) is used for gene biology functional analysis and sensitivity analysis of genes to pan cancer therapeutic drugs. Results TXN is highly expressed in most malignant tumors. The high expression of TXN is associated with overall survival (OS), disease-specific survival (DSS), disease-free interval (DFI), and progression free interval (PFI) in various cancers. Moreover, TXN expression is associated with TMB, MSI, tumor microenvironment, chemotherapy sensitivity and so on. Conclusion TXN may become a potential prognostic biomarker in pan cancer, providing strong theoretical basis for future tumor diagnosis and prognosis judgment. The retinoic acid-inducible gene-I (RIG-I)-like receptor signaling pathway, Toll-like receptor (TLR) signaling pathway, and nucleotide binding oligomerization domain (NOD)-like receptor signaling pathway may be crucial pathways through which TXN influences tumor immunity.
Humans ; Prognosis ; Neoplasms/diagnosis* ; Thioredoxins/metabolism* ; Microsatellite Instability ; Gene Expression Regulation, Neoplastic ; Biomarkers, Tumor/genetics* ; Mutation ; Tumor Microenvironment

The genome tagging project (GTP) plays a pivotal role in addressing a critical gap in the understanding of protein functions. Within this framework, we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411 (HA-tagged Ssp411) mouse model. This model is instrumental in probing the expression and function of Ssp411. Our research revealed that Ssp411 is expressed in the round spermatids, elongating spermatids, elongated spermatids, and epididymal spermatozoa. The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis. Nevertheless, rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions. Ssp411 is not detectable in metaphase II (MII) oocytes, zygotes, or 2-cell stage embryos, highlighting its intricate role in early embryonic development. These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP, fostering groundbreaking advancements in the fields of spermiogenesis and reproductive biology.
Animals ; Female ; Humans ; Male ; Mice ; Spermatids/metabolism* ; Spermatogenesis/physiology* ; Spermatozoa/metabolism* ; Thioredoxins/genetics*

The objective of this study was to construct an improved thioredoxin fusion protein expression system, and express the cathelicidin-derived peptide, Lf-CATH2. The improved fusion vector Lf-CATH2-pET32α(-TS) was successfully constructed by firstly deleting the thrombin site and S tag from the pET-32α vector, then inserting the Lf-CATH2 plus a thrombin site instead. Afterwards, Lf-CATH2 was expressed in Escherichia coli as fusion protein. After the cleavage by thrombin, Lf-CATH2 was released and subsequently separated using affinity chromatography. The antimicrobial activity of purified Lf-CATH2 was also examined. The improved expression vector significantly increased enzyme cleavage efficiency by 37%, and Lf-CATH2 could be expressed in high yield and maintain the biological activity. This novel thioredoxin fusion protein expression system enables a quick production of high-yield bioactive cationic peptides like cathelicidins.
Cathelicidins ; biosynthesis ; Chromatography, Affinity ; Escherichia coli ; Genetic Vectors ; Recombinant Fusion Proteins ; biosynthesis ; Thioredoxins ; genetics

Redox-regulating molecule, recombinant human thioredoxin (rhTRX) which shows anti-inflammatory, and anti-oxidative effects against lipopolysaccharide (LPS)-stimulated inflammation and regulate protein expression levels. LPS-induced reactive oxygen intermediates (ROI) and NO production were inhibited by exogenous rhTRX. We identified up/downregulated intracellular proteins under the LPS-treated condition in exogenous rhTRX-treated A375 cells compared with non-LPS-treated cells via 2-DE proteomic analysis. Also, we quantitatively measured cytokines of in vivo mouse inflammation models using cytometry bead array. Exogenous rhTRX inhibited LPS-stimulated production of ROI and NO levels. TIP47 and ATP synthase may influence the inflammation-related lipid accumulation by affecting lipid metabolism. The modulation of skin redox environments during inflammation is most likely to prevent alterations in lipid metabolism through upregulation of TIP47 and ATP synthase and downregulation of inflammatory cytokines. Our results demonstrate that exogenous rhTRX has anti-inflammatory properties and intracellular regulatory activity in vivo and in vitro. Monitoring of LPS-stimulated pro-inflammatory conditions treated with rhTRX in A375 cells could be useful for diagnosis and follow-up of inflammation reduction related with candidate proteins. These results have a therapeutic role in skin inflammation therapy.
Animals ; Antioxidants/*pharmacology ; Cell Line, Tumor ; Humans ; Inflammation/metabolism ; *Lipid Metabolism ; Lipopolysaccharides/pharmacology ; Mice ; Mice, Inbred C57BL ; Nitric Oxide/metabolism ; Proteome/genetics/metabolism ; Skin/drug effects/metabolism/pathology ; Thioredoxins/*pharmacology

Adenovirus transfection technique was used in the current study to show if thioredoxin-interacting protein (TXNIP) overexpression can induce cell apoptosis and injury in H9C2 cardiomyocytes cultured in normal glucose condition. And the mechanisms were then investigated. Briefly, H9C2 cardiomyocytes in logarithmic growth phase were randomly divided into three groups: normal cultured group, empty adenovirus vector group (Ad-eGFP) and TXNIP overexpression group (Ad-TXNIP-eGFP). All cells were cultured in DMEM containing normal concentration of glucose (5 mmol/L) and lipid. 72 h after adenovirus transfection, cells and culture mediums were collected for further assay. The results showed that Ad-eGFP and Ad-TXNIP-eGFP adenovirus transfected H9C2 cells successfully, and the transfection efficiency reached the peak at 72 h. Compared with Ad-eGFP group, Ad-TXNIP-eGFP transfection significantly increased TXNIP mRNA (P < 0.05) and protein expression level (P < 0.01). TXNIP overexpression induced remarkable cell apoptosis and injury as evidenced by increased caspase-3 activity (P < 0.05), apoptotic rate (P < 0.01) and LDH activity (P < 0.01). To further analysis the mechanisms of TXNIP-induced cell apoptosis, we also determined Trx activity, Trx related free radical injury and p38 kinase activation, which are involved in free radical induced apoptosis. The results showed that, compared with those in Ad-eGFP group, Trx activity was significantly decreased (P < 0.01), while malondialdehyde (MDA), 3-nitrotyrosine contents and p38 kinase activity were significantly increased (P < 0.01) in TXNIP overexpression group. These results suggest that TXNIP overexpression alone can induce severe apoptosis and injury in H9C2 cardiomyocytes even they are cultured in normal glucose and lipid concentration conditions. The mechanism involved is that overexpressed TXNIP can bind and inhibit Trx, impairs its antioxidative and antiapoptotic function, and then increases free radical induced injury and p38 kinase dependent apoptosis.
Adenoviridae ; genetics ; Animals ; Apoptosis ; Carrier Proteins ; genetics ; metabolism ; Caspase 3 ; metabolism ; Cell Line ; Genetic Vectors ; Myocytes, Cardiac ; cytology ; Rats ; Thioredoxins ; metabolism

Vitamin D3 up-regulated protein 1 (VDUP1) is a potent growth suppressor that inhibits tumor cell proliferation and cell cycle progression when overexpressed. In a previous study, we showed that VDUP1 knockout (KO) mice exhibited accelerated liver regeneration because such animals could effectively control the expression of cell cycle regulators that drive the G1-to-S phase progression. In the present study, we further investigated the role played by VDUP1 in initial priming of liver regeneration. To accomplish this, VDUP1 KO and wild-type (WT) mice were subjected to 70% partial hepatectomy (PH) and sacrificed at different times after surgery. The hepatic levels of TNF-alpha and IL-6 increased after PH, but there were no significant differences between VDUP1 KO and WT mice. Nuclear factor-kappaB (NF-kappaB), c-Jun-N-terminal kinase (JNK), and signal transducer and activator of transcription 3 (STAT-3) were activated much earlier and to a greater extent in VDUP1 KO mice after PH. A single injection of TNF-alpha or IL-6 caused rapid activation of JNK and STAT-3 expression in both mice, but the responses were stronger and more sustained in VDUP1 KO mice. In conclusion, our findings provide evidence that VDUP1 plays a role in initiation of liver regeneration.
Animals ; Blotting, Western ; Carrier Proteins/*genetics/metabolism ; Cell Proliferation ; *Gene Expression Regulation ; Hepatectomy ; Hepatocytes/*cytology/physiology ; JNK Mitogen-Activated Protein Kinases/genetics/metabolism ; Liver/*physiology ; Male ; Mice, Knockout ; NF-kappa B/genetics/metabolism ; Polymerase Chain Reaction ; *Regeneration ; STAT3 Transcription Factor/genetics/metabolism ; Thioredoxins/*genetics/metabolism

OBJECTIVE: To investigate the significance of human thioredoxin-2 (TRX-2) in monitoring minimal residual disease (MRD) in acute leukemia (AL). METHODS: We used real-time quantitative PCR to serially quantitize TRX-2 expression levels in the bone marrow of AL patients at diagnosis (n=68), at complete hematologic remission (CHR, n=57) and at relapse (n=25). Another 25 normal donors served as normal controls. The upper limit of the bone marrow at 91 was regarded as the reference. TRX-2 expression level at CHR with <5% blast cells in the bone marrow of relapse patients was analyzed and compared with MRD by flow cytometry. RESULTS: The TRX-2 levels between the CHR patients and newly diagnosed patients, and between the CHR patients and the relapse patients had significant difference. TRX-2 expression level of 21(21/25) relapse patients at CHR with <5% blast cells in the bone marrow was higher than the reference (>91). TRX-2 level was correlated to the expression level of MRD. CONCLUSION TRX-2 may be the marker for AL and used in MRD monitoring.
Case-Control Studies ; Female ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; metabolism ; Male ; Neoplasm, Residual ; diagnosis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Thioredoxins ; genetics ; metabolism

This study was purposed to construct and prepare the recombinant adenovirus vector carrying human thioredoxin (hTRX) and enhanced green fluorescence protein (EGFP), and transfect it into HEK293 cells, so as to lay a foundation for further gene therapy. The PCR-amplified products of hTRX with a pair of primers containing Not I and EcoR V restriction sites were subcloned into shuttle plasmid pDC316-mCMV. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid pDC316-hTRX-EGFP and large adenovirus-helper plasmid pBHGlox (delta) E1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus pAd-hTRX-EGFP was co-transfected in HEK293 cells, purified by CsCl gradient centrifugation, counted for virus particles and determined for titer. The recombinant adenovirus was identified by PCR. The HEK293 cells were then transfected with adenoviruses and assayed by flow cytometry. The expression of hTRX was confirmed by Western blot. The results showed that according to PCR and restriction endonuclease assay, the target gene was inserted into recombinant adenovirus vector successfully. The sequence of fusion gene was the same as that of designed fragments. The titer of the purified recombinant adenovirus pAd-hTRX-EGFP was 5.558×10(10) pfu/ml. A transfection efficiency of 92.25% could be achieved at MOI = 100. Western blot further confirmed that hTRX was efficiently expressed in HEK293 cells. It is concluded that recombinant adenovirus vector containing hTRX has been constructed successfully and obtained highly efficient virus that can express efficiently in HEK293 cells, which laid a foundation for further investigation.
Adenoviridae ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Recombinant Proteins ; genetics ; Thioredoxins ; genetics ; Transfection

OBJECTIVETo explore the role of oxidative stress and the antioxidant protein thioredoxin in the tumorigenesis and progression of gastric cancer.

METHODSThe plasma levels of adenosine deaminase (ADA), glutathione peroxidase (GPX), superoxide dismutase (SOD), and advanced oxidation protein products (AOPP) were determined by colorimetry, and the plasma levels of thioredoxin were determined by enzyme-linked immunosorbent assay (ELISA) in 48 gastric cancer patients and 30 healthy subjects. RT-PCR assay was employed to examine the expression levels of thioredoxin mRNA in the tissue samples of the patients.

RESULTSCompared with the healthy controls, patients with gastric cancer had significantly increased plasma levels of ADA and AOPP (P<0.05), decreased plasma GPX level (P<0.05), and similar plasma SOD levels. The plasma levels of thioredoxin were significantly higher in patients with gastric cancer than in the healthy controls (P<0.05). Thioredoxin levels was not associated with gender, age, degree of tumor cell differentiation, invasion depth, or lymph node metastasis (P>0.05), but was correlated to distant tumor metastasis (P<0.05). The expression of Trx mRNA was significantly higher in gastric carcinoma than in normal gastric tissue (P<0.05).

CONCLUSIONGastric cancer patients have high levels of oxidative stress and thioredoxin expression, and the latter is related to distant metastasis of the tumor.

Adenosine Deaminase ; blood ; Adult ; Advanced Oxidation Protein Products ; blood ; Aged ; Case-Control Studies ; Female ; Glutathione Peroxidase ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Oxidative Stress ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Superoxide Dismutase ; blood ; Thioredoxins ; genetics ; metabolism

Antimicrobial peptides (AMPs) are of great significance in the field of food, feed and medicine due to their wide spectrum of antimicrobial activity and new mechanism of action different from conventional antibiotics. AMPs production from natural sources is usually limited, and chemical synthesis is not economically practical, especially for the production of long peptides. Therefore, heterologous expression of AMPs has been widely studied as an alternative, and fusion expression plays an important role in increasing production. The present review mainly focuses on the types and bioactivities of AMPs. In addition, the recent strategies to the most commonly used carrier proteins for fusion expression of AMPs and prospects for future research were also discussed.
Anti-Infective Agents ; metabolism ; Antimicrobial Cationic Peptides ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Glutathione Transferase ; biosynthesis ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Thioredoxins ; biosynthesis ; genetics