Huaier inhibits cholangiocarcinoma cells through the twist1/FBP1/Wnt/ß-catenin axis.

BACKGROUND: Although Huaier granules can be used as prospective anti-cholangiocarcinoma drugs, the mechanism of action of Huaier granules in cholangiocarcinoma is not clear. The anti-cholangiocarcinoma effect of Huaier granules was validated in cell line research. In vitro experiments were conducted to investigate the signalling pathways affected by Huaier in CCA cells. METHODS AND RESULTS: Real-time quantitative PCR (RT‒qPCR) and Western blot analysis were performed to analyse gene expression in CCA cells. MTT assays, scratch tests, and Transwell assays were used to explore the effects on the proliferation and metastasis of CCA cells. Chromatin immunoprecipitation assays were performed to reveal the potential underlying mechanisms involved. Twist1 was upregulated in human CCA tissues. In addition, its expression levels were negatively related to FBP1 expression levels. Mechanistically, Twist1 can bind to the region of the FBP1 promoter to reduce its expression. Huaier plays an indispensable role in suppressing Twist1 expression to inhibit the Twist1/FBP1/Wnt/ß-catenin axis. Then, we verified the effect of Huaier in vitro. CONCLUSIONS: These findings suggested that Huaier granules were capable of inhibiting CCA development through regulating the Twist1/FBP1/Wnt/ß-catenin signalling axis and provided a novel orientation for the development of novel anti-CCA drugs.

Identification and Characterization of Sclerotium delphinii Causing Southern Blight on Aconitum kusnezoffii in Northeast China.

Aconitum kusnezoffii is a perennial medicinal plant that belongs to the Ranunculaceae family and is distributed mainly in Northeast and North China. In July 2018, a typical southern blight disease of A. kusnezoffii was observed in commercial fields of Qingyuan County, Fushun City, Liaoning Province, China. The fungus mainly infected stem base and tuberous roots of the plant by wrapping the hyphae and absorbing nutrition, resulting in tuberous root wilted or whole plant death. Morphological characteristics of colony and sclerotia of three representative strains isolated from the diseased plants differed from those of Sclerotium rolfsii isolated from A. carmichaelii. Sclerotia were large (0.8 to 5.1 mm), reddish-brown, and irregular and had pitted surfaces, and the hyphae were white, compact, or fluffy, with a growth rate ranging from 8.0 to 10.1 mm/day. Phylogenetic analysis of the internal transcribed spacer and the large subunit sequences of Akln6, Akln9, and Akln15 showed that three strains isolated from A. kusnezoffii formed a unique and well-supported clade that groups with the reference isolates of S. delphinii. Based on phylogenetic analysis and cultural and morphological characteristics, the three isolates of A. kusnezoffii were identified as S. delphinii. The optimum temperature for mycelial growth of the three tested isolates was 30°C, and sclerotia formed and matured more easily at 20°C. Light promoted the growth of mycelial, whereas dark was beneficial to the formation and maturation of sclerotia. The pathogenicity of S. delphinii showed stronger than S. rolfsii at low temperature (20°C). This is the first report of S. delphinii causing southern blight on A. kusnezoffii in China, and this finding provides a basis for disease-accurate diagnosis and the development of effective management strategies.

miR-328-3p, a Predictor of Stroke, Aggravates the Cerebral Ischemia-Reperfusion Injury.

BACKGROUND: In the present study, we aimed to identify microRNAs (miRNAs) that affected the prognosis of stroke and assess their biological effects. MATERIALS AND METHODS: A high-throughput sequencing (HTS) analysis was performed to screen distinctive miRNAs in serum exosomes of stroke patients, and these miRNAs were subsequently validated using individual quantitative real-time polymerase chain reaction (qRT-PCR) in a cohort consisting of 39 stroke patients and 20 normal controls. Briefly, miR-328-3p agomir or agomir NC was injected into rats before ischemia and reperfusion (I/R) injury. Zea-Longa score, neurological severity score (mNSS), triphenyltetrazolium chloride (TTC) staining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, transmission electron microscopy, and hematoxylin and eosin (H&E) staining were used to examine the brain injury. Immunohistochemistry was utilized to determine the expressions of TNF-α and IL-6. RESULTS: The expression of serum exosomal miR-328-3p was significantly reduced in patients with an infarct volume ≥10 cm3 (P=0.01). Serum exosomal miR-328-3p was associated with the short-term prognosis (P=0.02), and the level of miR-328-3p was an independent relative factor for short-term prognosis (OR 5.276, P=0.02). The sensitivity of miR-328-3p level higher than 1.24 to predict the severity of the patient's 1-week prognosis was 70%, and the specificity was 83% (AUC=0.74, P=0.02). The mNSS was higher in the miR-328-3p agomir group compared with the agomir NC group (P=0.03). Neutrophil infiltration was more serious in the miR-328-3p agomir group. CONCLUSION: Our study indicated that miR-328-3p played a critical predictive role in the short-term prognosis of stroke, and up-regulation of miR-328-3p aggravated cerebral I/R injury.