Henan University

As an essential biothiol, cysteine (Cys) is critical for regulating intracellular redox homeostasis. Dysregulated Cys levels correlated with various pathological conditions, establishing its value as a pathological biomarker. Drug-induced hepatocellular injury was frequently associated with aberrant fluctuations in intracellular Cys concentrations, highlighting the importance of developing highly specific Cys-responsive fluorescent probes for the early diagnosis and therapeutic monitoring of liver injury. In this study, a novel near-infrared (NIR) fluorescent probe BHC-A, was synthesized based on a dicyanomethylene-4H-benzopyran scaffold for the selective detection of Cys in both in vitro and in vivo models. BHC-A exhibited a large stokes shift (> 200 nm), high specificity and superior sensitivity to Cys, with a detection limit of 0.37 μM. The probe was successfully utilized for fluorescence imaging of Cys in live cells and enabled real-time monitoring of Cys fluctuations during acetaminophen-induced HepG2 hepatocyte injury. Furthermore, BHC-A facilitated in vivo visualization of Cys through NIR fluorescence imaging in animal models. Therefore, BHC-A provides represents a potential analytical tool for the early diagnosis of liver injury and high-throughput drug screening applications.

Malic acid markedly affects watermelon flavor. Reducing the malic acid content can significantly increase the sweetness of watermelon. An effective solution strategy is to reduce watermelon malic acid content through molecular breeding technology. In this study, we measured the TSS and pH of six watermelon varieties at four growth nodes. The TSS content was very low at 10 DAP and accumulated rapidly at 18, 26, and 34 DAP. Three phosphoenolpyruvate carboxykinase (PEPCK) genes of watermelon were identified and analyzed. The ClaPEPCK4 expression was inversely proportional to malate content variations in fruits. In transgenic watermelon plants, overexpressing the ClaPEPCK4 gene, malic acid content markedly decreased. In the knockout transgenic watermelon plants, two SNP mutations and one base deletion occurred in the ClaPEPCK4 gene, with the malic acid content in the leaves increasing considerably and the PEPCK enzyme activity reduced to half of the wild-type. It is interesting that the ClaPEPCK4 gene triggered the closure of leaf stomata under dark conditions in the knockout transgenic plants, which indicated its involvement in stomatal movement. In conclusion, this study provides a gene target ClaPEPCK4 for creating innovative new high-sweetness watermelon varieties.

This study aimed to investigate the association between CA9 expression and the immune infiltration and prognosis of CC.

The GSE9750 dataset in the GEO database, the UALCAN database, and the cBioPortal of cervical squamous cell carcinoma study were used for bioinformatic analysis, followed by validation via immunohistochemistry in clinical samples.

Compared to normal controls, CC samples showed 1175 significantly downregulated and 698 significantly upregulated genes, respectively. Specifically, CA9 was identified among the up-regulated genes, with verification by analysis of TCGA data. The overexpression of CA9 in CC was further confirmed in clinical samples. The protein-protein interaction network results revealed that CA9 was closely associated with 12 genes, which were widely involved in cancer-related processes such as cell proliferation, migration, and metabolism. CA9 was associated with tumor histological subtypes, hypoxia scores, low immune infiltration, poor survival rate, and specific microorganisms.

Our study indicated that CA9 was upregulated in CC and closely related to tumor immune microenvironment, thereby becoming an important prognostic factor for CC. This study highlights CA9 as a potential diagnostic and therapeutic target for the disease.