Synergistic impact of Gui Zhi Shao Yao Zhi Mu Decoction and leflunomide on gut microbiota in rheumatoid arthritis: insights from 16S rDNA sequencing.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic autoimmune disease with complex pathogenesis, including alterations in the gut microbiota. Gui Zhi Shao Yao Zhi Mu Decoction (GSZD), a traditional Chinese herbal formula, has shown efficacy in RA treatment, but its impact on intestinal microflora remains unclear. This study aimed to investigate the effects of GSZD combined with leflunomide on the gut microbiota of RA patients. METHODS: The study enrolled 48 RA patients who were randomly assigned to either a control group receiving leflunomide or a treatment group receiving GSZD combined with leflunomide for 12 weeks. Gut microbiota composition was analyzed pre- and post-intervention using 16S rDNA sequencing. Changes in microbial diversity, abundance, and metabolic functions were assessed. RESULTS: Post-treatment, both groups exhibited significant alterations in gut microbiota composition. GSZD combined with leflunomide led to an increased Bacteroidetes/Firmicutes ratio and a reduction in Actinobacteria compared to leflunomide alone. This was associated with beneficial shifts in microbial genera and metabolic pathways, suggesting improved gut health and systemic immune modulation. CONCLUSION: GSZD combined with leflunomide significantly modulates the gut microbiota in RA patients. This study provides insights into the mechanisms underlying the therapeutic effects of GSZD and highlights the potential of integrating traditional Chinese medicine with conventional treatments in managing RA.

Knowledge heterogeneity and corporate innovation performance: The mediating influence of task conflict and relationship conflict.

Innovation has emerged as a crucial factor in the sustenance and growth of enterprises. Nonetheless, small and medium-sized enterprises (SMEs) confront numerous challenges in their pursuit of innovation, owing to constraints in capital, expertise, and knowledge resources. Drawing on the resource-based theory and the input-process-output (IPO) model, this study devises a mechanism model to assess the impact of knowledge heterogeneity and innovation performance on small and medium-sized manufacturing enterprises in Guizhou Province, China. The objective is to offer recommendations for the advancement and innovation of enterprises with relative knowledge resource deficiencies. A total of 324 valid questionnaires were gathered, and the acquired data were analyzed employing SPSS 23.0 and Amos 26.0. The findings reveal that knowledge heterogeneity exerts a significantly positive influence on innovation performance. Task conflict and relationship conflict serve as partial mediators in the effects of knowledge heterogeneity on innovation performance. By capitalizing on the heterogeneity of internal and external knowledge, enterprises can effectively enhance their innovation outcomes. Furthermore, the study demonstrates that knowledge sharing possesses a moderating effect on the impact of knowledge heterogeneity on task conflict, relationship conflict, and innovation performance. In a conducive sharing environment, the ultimate effect of knowledge heterogeneity on innovation is subject to alteration.

Functional characterization of proton antiport regulation in the thylakoid membrane.

During photosynthesis, energy is transiently stored as an electrochemical proton gradient across the thylakoid membrane. The resulting proton motive force (pmf) is composed of a membrane potential (ΔΨ) and a proton concentration gradient (ΔpH) and powers the synthesis of ATP. Light energy availability for photosynthesis can change very rapidly and frequently in nature. Thylakoid ion transport proteins buffer the effects that light fluctuations have on photosynthesis by adjusting pmf and its composition. Ion channel activities dissipate ΔΨ, thereby reducing charge recombinations within photosystem II. The dissipation of ΔΨ allows for increased accumulation of protons in the thylakoid lumen, generating the signal that activates feedback downregulation of photosynthesis. Proton export from the lumen via the thylakoid K+ exchange antiporter 3 (KEA3), instead, decreases the ΔpH fraction of the pmf and thereby reduces the regulatory feedback signal. Here, we reveal that the Arabidopsis (Arabidopsis thaliana) KEA3 protein homo-dimerizes via its C-terminal domain. This C-terminus has a regulatory function, which responds to light intensity transients. Plants carrying a C-terminus-less KEA3 variant show reduced feed-back downregulation of photosynthesis and suffer from increased photosystem damage under long-term high light stress. However, during photosynthetic induction in high light, KEA3 deregulation leads to an increase in carbon fixation rates. Together, the data reveal a trade-off between long-term photoprotection and a short-term boost in carbon fixation rates, which is under the control of the KEA3 C-terminus.

Development of a highly thermostable immunoassay based on a nanobody-alkaline phosphatase fusion protein for carcinoembryonic antigen detection.

Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM-5) assays are employed in routine clinical settings to diagnose tumor. We selected two nanobodies with high-affinity to CEACAM-5, termed Nb11C12 and Nb2D5, using phage-display technology. The Nb2D5 fused with calf intestinal alkaline phosphatase (CAP), human placental alkaline phosphatase (HAP), or Pyrococcus abyssi alkaline phosphatase (PAP) were expressed in human embryonic kidney (HEK293) cells. The enzymatic activity of Nb2D5-HAP fusion protein was the best and remained stable at 60 °C for 7 days. The affinity of Nb2D5-HAP fusion protein to CEACAM-5 reached 42 pM. A chemiluminescent enzyme immunoassay (CLEIA) based on Nb2D5-HAP fusion protein was established for quantitative CEACAM-5 assay in clinical settings. The CLEIA exhibited a wide linear range of 0.31-640 ng/mL toward CEACAM-5, with a limit of detection (LOD) of 0.85 ng/mL. No cross-reactivity occurred with CEACAM-1, CEACAM-3, CEACAM-6, or CEACAM-8, and no interference was observed with rheumatoid factors. The CLEIA based on Nb2D5-HAP fusion protein was stable for 8 weeks at 37 °C and 50% relative humidity. The CLEIA developed from Nb2D5-HAP fusion protein had much better stability and linearity with similar reproducibility compared with the enzyme-linked immunosorbent assay developed from conventional monoclonal antibodies, which have been widely used in clinics over the past several decades. Graphical abstract.

Shenqi Fuzheng Injection (SFI) Enhances IFN-α Inhibitory Effect on Hepatocellular Carcinoma Cells by Reducing VEGF Expression: Validation by Gene Silencing Technique.

Shenqi Fuzheng Injection (SFI) is a traditional Chinese medicine injection with anticancer properties and is mainly composed of ginseng and astragalus. Its efficacy has been confirmed in clinical trials, but the mechanism remains unclear. We investigated the effect of SFI on vascular endothelial growth factor (VEGF) gene expression in hepatocellular carcinoma (HCC) cells and identified its possible mechanism of synergistic effects when combined with the chemotherapeutic drug interferon (IFN-) α. An MTT assay was used to measure the inhibition effects of low-dose IFN-α (6000 IU) with or without SFI (0.5 g/L) on the HCC cell line MHCC97. VEGF-silenced MHCC97L-mir200 cell lines were prepared using lentiviral vectors and evaluated by real-time PCR to determine the inhibition effect. We examined MHCC97L-mir200 and MHCC97L cells by MTT assay, using IFN-α alone or in combination with SFI. The inhibition ratio of IFN-α (6000 IU) was -29.5%, while that for IFN-α (6000 IU) + SFI (0.5 g/L) was 17.0%, which was significantly higher than that for the IFN-α group (P < 0.01). The VEGF gene was silenced successfully in MHCC97-L cells. After interference of VEGF, the inhibition by SFI and IFN-α in MHCC97L-mir200 did not differ from that in MHCC97-L cells (P > 0.05). SFI can reduce the expression of VEGF in HCC, which can increase the efficacy of IFN-α, providing a theoretical basis for clinical application.

Immuno-Efficacy of a T. gondii Secreted Protein with an Altered Thrombospondin Repeat (TgSPATR) As a Novel DNA Vaccine Candidate against Acute Toxoplasmosis in BALB/c Mice.

Toxoplasma gondii (T.gondii) is distributed worldwide and infects most species of warm-blooded animals, including humans. Toxoplasmosis has serious consequences, especially in people with an impaired or immature immune system. Thus, an effective vaccine is urgently required. Secretory microneme proteins are essential for the adhesion and invasion of T. gondii. The gene encoding the microneme protein, T. gondii secreted protein with an altered thrombospondin repeat (TgSPATR), we constructed a recombinant eukaryotic plasmid, pVAX1-TgSPATR, as a DNA vaccine, injected it intramuscularly into BALB/c mice and evaluated the induced immune response. Lymphocyte proliferation assays, cytokine (IFN-γ, IL-2, IL-4, IL-10), and antibody determinations showed that mice immunized with pVAX1-TgSPATR produced humoral and mixed Th1/Th2 type cellular immune responses. The survival times of mice immunized with pVAX1-TgSPATR were also significantly prolonged (15.7 ± 1.42 days) compared with control groups, which died within 7 days of challenge (p < 0.05). The current study indicated that pVAX1-TgSPATR induce a T. gondii specific immune response and might be a promising vaccine candidate against toxoplasmosis. To the best of our knowledge, this is the first report to evaluate the immunoprotective value of TgSPATR against T. gondii.

Genome-wide miRNAs expression profiles of Schistosoma japonicum schistosomula in response to artesunate.

AIM: miRNAs play a significant role in pharmacogenomics and are likely to be important in the molecular mechanism of atesunate (ART) effects on Schistosoma japonicum. METHODS: We sequenced the RNAs using an Illumina (Solexa) DNA sequencer and compared the relative expression levels of the miRNAs in 10-day-old schistosomula from ART and the parallel control group. RESULTS: We characterized 95 known miRNAs from S. japonicum schistosomula individuals, including 38 novel miRNA families. Among the detectable 134 miRNAs differentially expressed (>2.0-fold change, p < 0.01) after ART treatment in schistosomula, a total of seven known or novel 3p- or 5p- derived S. japonicum miRNAs were characterized. We propose that sja-miR-125b may regulate the expression of ART metabolizing enzymes, glutathione synthetase or heme-binding protein 2 to help S. japonicum resists or adapts to drug stress and also ART may significantly inhibit sexual maturation of female worms mediated by mir-71b/2 miRNA cluster. CONCLUSION: This was the first comprehensive miRNAs expression profile analysis of S. japonicum in response to ART, and provides an overview of the complex network of the mechanism of action of ART on S. japonicum.

[Application of Nano Carbon-based Immunosensor in Pathogen Detection].

The biosensors exhibit many advantages such as simple operation, rapid reaction and high sensitivity in pathogen detection. The sensitivity and specificity of the biosensors can be significantly enhanced by the combined use of carbon nano-materials (such as carbon nanotubes and graphene) and bio-sensing devices. This paper reviews the characteristics of carbon nano-biosensors, its applications in pathogen detection and new development.

Quantitative proteomic analyses of Schistosoma japonicum in response to artesunate.

Artesunate (ART) has high prophylactic efficacy against Schistosoma japonicum infections and has been used to treat and prevent schistosomiasis in China since 1995. However, the molecular mechanism of ART's effects on S. japonicum remains unclear. Herein, we applied isobaric tagging reagents for relative and absolute quantification analyses coupled with two-dimensional liquid chromatography and tandem mass spectrometry to investigate the effect of ART on the proteome of S. japonicum in susceptible mice. 4529 proteins were quantified on the basis of 21,825 unique peptides. Comparative proteomic analyses revealed that 145, 228 and 185 proteins were significantly differentially expressed after ART treatment in schistosomula, juvenile and adult worms, respectively. Ninety proteins were differentially expressed between each two treatment groups in response to ART treatment: 67 proteins were associated with S. japonicum development/aging and 23 were specifically associated with ART treatment. Quantitative real-time PCR of selected genes verified the proteomic data. Gene ontology annotation and Kyoto encyclopedia of genes and genomes pathway mapping analysis showed that the majority of differentially expressed proteins were involved in stress/defense/detoxification, signal transduction, carbohydrate metabolism, amino acid metabolism, transcription/translation, and protein synthesis/assembly/degradation. Thirty-four of the proteins differentially expressed under ART treatment encoded hypothetical, uncharacterized proteins with unknown functions. This study obtained the first comprehensive protein expression profile of S. japonicum in response to ART, and provides a basis for a better understanding of the molecular mechanisms of ART effects on S. japonicum.