Applied Biochemistry and Biotechnology最新文献

Hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC) remains a major global health challenge due to late diagnosis and high tumor heterogeneity. Cytokines, as key mediators of the tumor microenvironment, contribute to immune evasion and tumor progression in HBV-HCC. However, their prognostic significance remains unclear. Bulk RNA sequencing revealed differentially expressed cytokine genes in HBV-HCC. A cytokine-based gene signature was created using the LASSO model, and Single-cell RNA sequencing analyzed gene expression at the cellular level. Serum and in vitro assays confirmed the key cytokine's role in tumor progression. Ten cytokine genes were differentially expressed in HBV-HCC, and six (CCL19, CCL20, CXCL2, GHR, IL1RAP, and LIFR) were selected to construct the LASSO-based risk model. This six-cytokine signature stratified patients into high- and low-risk groups with significantly different overall survival (P < 0.0001), achieving an area under the curve (AUC) exceeding 0.7 for 3-year survival in both training and validation cohorts. scRNA-seq revealed distinct cellular expression patterns of these cytokines, providing insights into their roles in tumor heterogeneity. High-risk patients exhibited enriched cell proliferation pathways and pronounced immunosuppression, whereas low-risk patients were associated with metabolic pathways. Drug sensitivity analysis identified 61 differentially responsive antitumor agents between risk groups. Notably, serum GHR levels increased during fibrosis but declined significantly in HBV-HCC. Functional assays demonstrated that GHR overexpression suppressed proliferation, migration, and invasion while promoting apoptosis. Our study integrates bulk and single-cell transcriptomics with functional validation, unveiling cytokine-driven mechanisms in HBV-HCC. The cytokine-based prognostic model holds promise for risk stratification, immunomodulation, and personalized therapy, offering new avenues for improving HBV-HCC management.

Cerebrovascular accidents, particularly ischemic stroke, constitute a paramount global health burden. While Salidroside (Sal), extracted from Rhodiola rosea, shows promise for neuroprotection, the specific molecular pathways mediating these effects, particularly the role of long non-coding RNA PVT1 and its interaction with miR-384-5p, remain unexplored. This investigation sought to fill this gap by deciphering how Sal influences cerebral ischemic pathology through the PVT1/miR-384-5p regulatory axis. Using murine middle cerebral artery occlusion (MCAO) and microglial BV2 cell oxygen-glucose deprivation (OGD) models, we evaluated neurological function, tissue damage, cellular viability, and molecular markers. We examined interactions between long non-coding RNA PVT1 and miR-384-5p through molecular techniques including dual-luciferase assays and RNA immunoprecipitation, while assessing oxidative parameters and inflammatory mediators. Sal administration (100 mg/kg/day) markedly improved neurological outcomes (35% reduction in deficit scores, p < 0.01) and diminished infarct dimensions by 42% in MCAO mice. Histological examination revealed preservation of neural architecture with 65% reduction in neuronal damage rate and 58% decrease in TUNEL-positive apoptotic cells in treated animals. At the cellular level, Sal preserved microglial viability (68% survival vs. 41% in OGD alone) while reducing reactive oxygen species generation by 52% and pro-inflammatory cytokine production (TNF-α by 61%, IL-1β by 55%, IL-6 by 48%). Importantly, cerebral ischemia elevated PVT1 expression 3.2-fold, which Sal effectively counteracted. Rescue experiments demonstrated that PVT1 overexpression abolished the protective benefits conferred by Sal treatment, confirming the functional significance of the PVT1/miR-384-5p regulatory circuit. Sal mitigates cerebral ischemic injury through disruption of the PVT1/miR-384-5p regulatory circuit, providing both mechanistic insights and a potential therapeutic strategy for ischemic stroke intervention. These findings establish specific molecular targets for pharmacological development in stroke therapy.

Dactylicapnos scandens (D. Don) Hutch is popular among traditional healthcare providers in Nagaland, North-East India and Bai communities in China. Current investigation analyses traditional claims of Dactylicapnos scandens root tuber extract to control diabetes through down-regulation of hepatic gluconeogenesis and oxidative stress in diabetes mellitus (Type 2) in both in vitro (cell culture) and in vivo (animal model) systems. Potential of Dactylicapnos scandens against diabetes was demonstrated in free fatty acid (FFA)-induced CC1 liver cells and streptozotocin-induced diabetic male Swiss albino mice. Enriched bioactive fraction was prepared through activity guided fractionation and major component was identified to be protopine. Enriched fraction of Dactylicapnos scandens root tuber and its isolated compound, protopine increased glucose uptake by the liver cells in FFA-induced condition. Further, the enriched fraction down regulated gluconeogenesis enzymes (Glucose-6-phosphatase and phosphoenolpyruvate kinase) through AMP-activated protein kinase (AMPK) modulation in both CC1 liver cells and liver tissues of diabetic mice. Moreover, in vivo experiment with enriched fraction containing protopine enhanced the tolerance of glucose as well as insulin level in plasma. Antidiabetic activity of enriched fraction containing protopine as a major component was also supported by histopathological analysis. Overall, protopine containing root tuber enriched fraction of Dactylicapnos scandens regulates antihyperglycemic efficacy in CC1 liver cells and in Swiss albino mice induced with diabetes via ameliorating AMPK/PEPCK/G6Pase signalling pathway.

Ultraviolet (UV) radiation accelerates skin aging and carcinogenesis, driving the demand for safer sunscreens. This study evaluated the SPF-enhancing potential of Hippophae rhamnoides (Elaeagnaceae) and Sapindus mukorossi (Sapindaceae) extracts in octocrylene-based oil-in-water emulsions. Formulations with 5% octocrylene ± 5% herbal extracts were assessed for in vitro SPF, antioxidant activity, and stability. The Hippophae rhamnoides and octocrylene combination achieved the highest SPF (15.42 ± 0.60), a 2.4-fold increase over octocrylene alone (6.47 ± 0.74), while Sapindus mukorossi yielded 9.03 ± 0.65. Antioxidant assays showed superior radical scavenging for Hippophae rhamnoides formulations (IC₅₀: 18.4 µg/mL) compared with Sapindus mukorossi (29.6 µg/mL). All formulations remained physically and chemically stable over 8 weeks. These results demonstrate that Hippophae rhamnoides can markedly enhance photoprotection and reduce synthetic filter load, supporting the development of hybrid herbal-synthetic sunscreens. In vivo validation and photostability testing are recommended.

Green synthesis is a method in which metal nanoparticles (MNPs) are produced by using biological synthesizing agents, including plant extracts. Copper nanoparticles (CuNPs) are used in a wide range of biological and non-biological sciences due to their unique physical and chemical properties. The research aim is to investigate the potential of producing CuNPs with cinnamon plant extract. Synthesis of CuNPs was done with the hydroalcoholic extract of the cinnamon plant. The Tyndall effect, ultraviolet-visible (UV-Vis), dynamic light Scattering (DLS), fourier transform infrared spectroscopy (FTIR), zeta potential (Zeta-P), raman spectroscopy, field emission scanning electron microscopy (FE-SEM), atomic force microscope (AFM), and x-ray diffraction (XRD) were used for evaluating CuNPs. Cytotoxic effects of nanoparticles (NPs) were measured. The average size of CuNPs is 40 nm and spherical, homogeneous, with uniform particle size. The highest cytotoxicity of NPs occurred after 48 h. Green synthesis of NPs using edible and medicinal plants such as cinnamon i.e. a suitable option due to its low cost, environmental friendliness, and easy process. In this research, the results indicate that the synthesized CuNPs have strong cytotoxic effects.

Tumor-associated macrophages (TAMs) differentiate into two main types based on signals from their microenvironment: the classically activated M1 type and the alternatively activated M2 type. An increased presence of M2-type TAMs has been identified in colorectal cancer patients, and the laminin alpha-1 chain (LAMA1), a key structural component of the extracellular matrix, plays a pivotal role in tumor metastasis. However, the exact mechanism by which LAMA1 promotes the polarization of TAMs towards the M2 phenotype and induces colorectal cancer remains elusive. In this work, we investigated the levels of LAMA1 and M2 TAMs in peripheral serum and tissue specimens from patients with colorectal cancer by utilizing Western Blotting, ELISA, and multicolor immunofluorescence. Peripheral blood samples from 20 patients were collected for ELISA analysis, with samples from 5 healthy individuals serving as controls. Additionally, tissue samples from 5 tumor patients and 5 normal controls were used to assess LAMA1 protein levels. Furthermore, statistical methods were employed to analyze the relationship between LAMA1 levels, M2 TAMs infiltration, and the pathological characteristics of colorectal cancer. Then we utilized Western Blotting, qPCR, and immunofluorescence to detect M2 TAMs markers and polarization mechanism after treatment with exogenous LAMA1 protein. Moreover, we established a mouse subcutaneous tumor model and utilized Western Blotting, immunohistochemistry and multicolor immunofluorescence to assess LAMA1 levels, tumor proliferation and proportion of M2 TAMs. Our analysis revealed that LAMA1 is overexpressed in the tumor microenvironment (TME) of colorectal cancer patients, which correlates with increased infiltration of M2-type TAMs. Furthermore, we found that the EGFR/AKT/CREB signaling pathway contributes to the polarization of TAMs toward the M2 subtype. Both in vitro and in vivo experiments demonstrated that LAMA1 promotes M2 macrophage polarization and facilitates tumor growth. Overall, these findings highlight a central role for LAMA1 in regulating macrophage polarization through the EGFR/AKT/CREB signaling pathway. Consequently, this process contributes to immune suppression and promotes tumor progression.

Although N6-methyladenosine (m6A) demethylase alkylation repair homolog protein 5 (ALKBH5) is closely associated with the aggressiveness of head and neck squamous cell carcinoma (HNSCC), the underlying molecular mechanisms remain to be fully elucidated. In this study, we screened out a novel ALKBH5/lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1)/miR-654-3p/HOXA10 axis that participated in the regulation of HNSCC development. Specifically, lncRNA NEAT1 was found to be highly expressed in HNSCC tissues and cells, and deletion of NEAT1 suppressed cell proliferation and tumorigenesis, and promoted apoptosis and autophagy to hamper cancer progression in HNSCC. Mechanistic experiments verified that the stability and expressions of NEAT1 were positively regulated by ALKBH5-mediated m6A demethylation, and ALKBH5 deletion induced a suppressive effect on HNSCC cell malignancies were all abrogated by overexpressing NEAT1. In addition, we evidenced that NEAT1 promoted Homeobox A10 (HOXA10) expression via sponging miR-654-3p in a competing endogenous RNA (ceRNA)-dependent manner, and both miR-654-3p inhibition and HOXA10 overexpression recovered cancer malignancies in the NEAT1-deficient HNSCC cells. Taken together, we concluded that ALKBH5-induced m6A demethylation increased the stability and expression levels of NEAT1, and elevated NEAT1 facilitated cancer progression in HNSCC through regulating the downstream miR-654-3p/HOXA10 axis. This study provided potential biomarkers for the diagnosis, treatment and prognosis of HNSCC in clinic.

Polyvinyl alcohol (PVA) is a well-known packaging material; however, its major drawback is the lack of inherent antibacterial properties, which are essential for preventing food spoilage. In the present work, we developed a method to enhance the antibacterial properties of PVA thin films by incorporating nanoparticles (NPs) into the films. Undoped and cobalt (Co)-doped zinc oxide (ZnO) nanoparticles were synthesized using PVA as a capping agent. The nanoparticles were prepared via chemical precipitation and microwave-assisted methods, and characterized using UV-Visible spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and X-Ray Diffraction (XRD). The crystallite sizes of the undoped and Co-doped ZnO were found to be 30.6 nm and 22.9 nm, respectively. SEM imaging revealed variations in surface morphology with different concentrations of PVA capping. PVA thin films containing 0.1%, 0.2%, and 0.3% of the synthesized nanoparticles were prepared and evaluated for their antimicrobial activity against Gram-negative bacteria Escherichia coli (MTCC 1559) and Gram-positive bacteria Enterococcus faecalis (MTCC 439). The results demonstrated that the incorporation of nanoparticles significantly enhanced the antibacterial activity of the PVA films.