The gut microbiome has been recognized as a significant contributor to primary hepatocellular carcinoma (HCC), with mounting evidence indicating associations between bacterial components and cancers of the digestive system. Here, to characterize gut bacterial signature in patients with primary HCC and to assess the diagnostic potential of bacterial taxa for primary HCC, 21 HCC patients and 21 healthy first-degree relatives (control group) were enrolled in this study. Bacterial DNA in the fecal samples was quantified by 16S rRNA gene sequencing. We found that that 743 operational taxonomic units (OTUs) were shared between patients with primary HCC and healthy controls. Of these, 197 OTUs were unique to patients with primary HCC, while 95 OTUs were unique to healthy subjects. Additionally, we observed significant differences in the abundance of Ruminococcaceae_UCG-014 and Romboutsia between patients with primary HCC and their healthy first-degree relatives. Besides, the relative abundance of Ruminococcaceae_UCG-014, and Prevotella_9 was positively correlated with physiological indicators including AST, ALT, ALB, or TBIL. Signature bacterial taxa could serve as non-invasive biomarkers, of which Romboutsia and Veillonella were identified as differential taxa in fecal samples from patients with HCC compared to healthy controls. Romboutsia showed a strong association with HCC (AUC = 0.802). Additionally, the combination of Romboutsia and Veillonella (AUC = 0.812) or the grouping of Fusobacterium, Faccalibacterium, and Peptostreptococcacae together (AUC = 0.762) exhibited promising outcomes for the diagnosis of HCC. The composition of gut microbes in patients with HCC was found to be significantly altered. Differential taxa Romboutsia, Veillonella, and Peptostreptococcacae could be tested for identification of HCC.

A method for the quantitative determination of eight polybrominated diphenyl ethers (PBDEs) in human milk was established using gas chromatography-negative chemical ionization/mass spectrometry (GC-NCI/MS). After extraction using the Soxhlet extractor, the lipid in the extract was removed by acidic silica, and the analytes were further purified by the multiplayer silica-alumina column. The concentrations of BDE28, BDE47, BDE99, BDE100, BDE153, BDE154, BDE183, and BDE209 in human milk were quickly determined by GC-NCI/MS, in selected ion monitoring (SIM) mode on a 7 m column. The quantitative analysis was performed using the isotope dilution method for BDE209 and internal standard method for other PBDEs. The limits of detection (LODs) of PBDEs in the lipid ranged from 1.74 to 6.35 pg/g. The recoveries of the spiked PBDEs were between 61.5% and 108%, and their relative standard deviations (RSDs) (n = 6) were between 2.06% and 10.1%. The method was further validated by the analysis of the reference materials of human milk. It was demonstrated to be a rapid, economical and reliable alternative for the trace analysis of selected PBDEs, especially for BDE209 in human milk.

The title compound C₁₉H₁₇F₃N₆ was synthesized and structurally characterized by infrared and mass spectroscopy, ¹H NMR, elemental analyses and single crystal X-ray diffraction. The compound crystallizes in monoclinic system, space group P2₁/c with a = 17.097(4) Å, b = 7.1668(16) Å, c = 18.389(3) Å, β = 118.251(15)°, V = 1984.8(8) ų, Z = 4, D_c = 1.293 g cm^–3, F(000) = 800, μ(MoK_α) = 0.10 mm^–1, R₁ = 0.0667, and wR₂ = 0.2084 for reflections with I > 2σ(I). Pyrazolo[1,5-a]pyrimidine and phenyl ring are almost coplanar, and the piperazine ring is in a chair conformation. The crystal structure is stabilized by C–H⋅⋅⋅N hydrogen interactions and a number of weak π⋅⋅⋅π interactions. In addition, the results of the determination of biological activity showed that the compound exhibited significant inhibitory activity against K562 and MKN45 cancer cell lines.

Stress granules (SGs) are highly dynamic and reversible cytoplasmic biomolecular condensates formed via liquid‐liquid phase separation (LLPS) under various stresses. As inherently heterogeneous assemblies, SGs possess distinct stable cores (initial nucleation seeds), substructures, or microphases. However, the mechanisms governing the formation and heterogeneity of SG nucleation seeds, and their dynamic integration, remain largely unclear. Here, we demonstrate that LENG8 is recruited to SGs under multiple stress conditions and is indispensable for SG assembly. Upon stress exposure, nuclear LENG8 granules disassemble, enabling LENG8 to translocate into the cytoplasm and undergo LLPS to form independent initial nucleation foci distinct from canonical G3BP1/TIA1‐dependent seeds. Subsequently, these LENG8‐initiated foci merge into growing SGs through a direct interaction between the prion‐like domain of LENG8 and TIA1, facilitating SG expansion and maturation. Depletion of LENG8 or disruption of the LENG8‐TIA1 interaction markedly impairs SG formation. Using conditional Leng8 knockout mice, we further establish that LENG8 deficiency attenuates stress‐induced SG assembly and increases cellular apoptosis in germ cells. Collectively, our study identifies LENG8 as a previously unrecognized SG nucleator, revealing the hierarchical assembly and integration mechanism of distinct nucleation modules during early SG biogenesis.

Spermatogenesis is a metabolically intensive process that is highly sensitive to perturbations in proteostasis. The integrated stress response (ISR) and its central effector, ATF4, orchestrate adaptive responses to maintain cellular homeostasis under stress; however, the functional significance of ATF4 in mammalian spermatogenesis has not been established. To investigate this, we engineered a conditional knockout mouse model with germ cell-specific deletion of the Atf4 gene. Results showed that Atf4 deletion did not impair spermatogenesis or male fertility, with knockout mice exhibiting normal testicular histology and standard sperm parameters. Proteomic analysis, however, revealed that ATF4 contributes to testicular protein expression homeostasis, as its deficiency caused marked dysregulation of the testicular proteome, especially impacting SQSTM1/p62 downregulate through endoplasmic reticulum (ER) stress pathway. We conclude that ATF4’s role in regulating proteostatic balance is functionally decoupled from its necessity for the core progression of spermatogenesis. These findings define ATF4 as a potential resilience agent safeguarding testicular function under ER stress, rather than a direct regulator of male germ cell development.