BACKGROUND: Temozolomide (TMZ) resistance is a significant challenge in treating glioblastoma (GBM). Collagen remodeling has been shown to be a critical factor for therapy resistance in other cancers. This study aimed to investigate the mechanism of TMZ chemoresistance by GBM cells reprogramming collagens. METHODS: Key extracellular matrix components, including collagens, were examined in paired primary and recurrent GBM samples as well as in TMZ-treated spontaneous and grafted GBM murine models. Human GBM cell lines (U251, TS667) and mouse primary GBM cells were used for in vitro studies. RNA-sequencing analysis, chromatin immunoprecipitation, immunoprecipitation-mass spectrometry, and co-immunoprecipitation assays were conducted to explore the mechanisms involved in collagen accumulation. A series of in vitro and in vivo experiments were designed to assess the role of the collagen regulators prolyl 4-hydroxylase subunit alpha 1 (P4HA1) and yes-associated protein (YAP) in sensitizing GBM cells to TMZ. RESULTS: This study revealed that TMZ exposure significantly elevated collagen type I (COL I) expression in both GBM patients and murine models. Collagen accumulation sustained GBM cell survival under TMZ-induced stress, contributing to enhanced TMZ resistance. Mechanistically, P4HA1 directly binded to and hydroxylated YAP, preventing ubiquitination-mediated YAP degradation. Stabilized YAP robustly drove collagen type I alpha 1 ( COL1A1) transcription, leading to increased collagen deposition. Disruption of the P4HA1-YAP axis effectively reduced COL I deposition, sensitized GBM cells to TMZ, and significantly improved mouse survival. CONCLUSION P4HA1 maintained YAP-mediated COL1A1 transcription, leading to collagen accumulation and promoting chemoresistance in GBM.
Temozolomide ; Humans ; Glioblastoma/drug therapy* ; Animals ; Mice ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics* ; YAP-Signaling Proteins ; Hydroxylation ; Dacarbazine/pharmacology* ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Collagen/biosynthesis* ; Collagen Type I/metabolism* ; Prolyl Hydroxylases/metabolism* ; Antineoplastic Agents, Alkylating/therapeutic use*

Threatened abortion is a common disease of obstetrics and gynecology and one of the diseases responding specifically to traditional Chinese medicine （TCM）. The China Association of Chinese Medicine organized experts in TCM obstetrics and gynecology， Western medicine obstetrics and gynecology， and pharmacology to deeply discuss the advantages of TCM and integrated Chinese and Western medicine treatment as well as the medication plans for threatened abortion. After discussion， the experts concluded that chromosome， endocrine， and immune abnormalities were the key factors for the occurrence of threatened abortion， and the Qi and blood disorders in thoroughfare and conception vessels were the core pathogenesis. In the treatment of threatened abortion， TCM has advantages in preventing miscarriages， alleviating clinical symptoms and TCM syndromes， relieving anxiety， regulating reproductive endocrine and immune abnormalities， personalized and diversified treatment， enhancing efficiency and reducing toxicity， and preventing the disease before occurrence. The difficulty in diagnosis and treatment of threatened abortion with traditional Chinese and Western medicine lies in identifying the predictors of abortion caused by maternal factors and the treatment of thrombophilia. Recurrent abortion is the breakthrough point of treatment with integrated traditional Chinese and Western medicine. It is urgent to carry out high-quality evidence-based medicine research in the future to improve the modern diagnosis and treatment of threatened abortion with TCM.

Metagenomic cosmid libraries containing 1.26 x 10(5) clones, covering about 4.8 x 10(6) kb metagenomic DNA of uncultured microorganisms from the contents of buffalo rumens were constructed, and 118 independent clones expressing beta-glucosidase activity were isolated from the libraries. Screening of these clones showed that eight clones expressed relatively higher beta-glucosidase activity at pH 5.0 and 37 degrees C. One out of the eight clones was subcloned. Sequencing analysis showed that an open reading frame (ORF) of 2223 bp, termed umcel3G, potentially encodes a beta-glucosidase. The encoded product shared highest homology with a beta-glucosidase from Bacillus sp. at 60% identity and 73% similarity. The umcel3G was over-expressed in Escherichia coli and the size of the translated product Umcel3G on SDS-PAGE was in agreement with the predicted molecular mass. Zymogram analysis showed that Umcel3G exhibited beta-glucosidase activity, confirming that this ORF encodes a beta-glucosidase. The Umcel3G, purified with Ni-NTA column, exhibited optimal activity at pH 6.0-6.5 and 45 degrees C. Certain ions such as Ca2+, Zn2+ had significant positive effect on the activity of Umcel3G. However, some ions such as Fe3+, Cu2+ gave significant inhibitory effect on the enzyme. The Ni-NTA purified recombinant beta-glucosidase Umcel3G had a specific activity of 22.8 IU/mg at pH4.5, 35 degrees C and at the presence of 5 mmol/L Ca2+, indicating that this enzyme has potential applications in the fermentative production of ethanol by simultaneous saccharification and cofermentation (SSCF) of lignocelluloses.
Animals ; Bacteria ; enzymology ; genetics ; Buffaloes ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Open Reading Frames ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Rumen ; microbiology ; beta-Glucosidase ; biosynthesis ; genetics ; isolation & purification