Matrine protects against DSS-induced murine colitis by improving gut barrier integrity, inhibiting the PPAR-α signaling pathway, and modulating gut microbiota.

Matrine is a naturally occurring quinolizidine alkaloid with various bioactivities. However, little is known of its function on ulcerative colitis (UC). Here, we investigated the effect and underlying mechanisms of matrine on dextran sulfate sodium (DSS)-induced UC mice. In this study, different concentrations of matrine were given to mice with DSS-induced colitis for a week. The symptoms of colitis, colonic pathology, inflammation-related indicators, and intestinal mucosal barrier function were detected and analyzed. Moreover, RNA-seq analysis in colon tissues was conducted, and 16S rDNA sequencing was carried out to evaluate the gut microbiota of colon contents. The results showed that matrine significantly alleviated clinical activity and histological changes of UC mice, inhibited the production of the pro-inflammatory cytokines, and improved gut barrier integrity. Moreover, RNA-seq analysis and experimental verification showed that matrine significantly inhibited the peroxisome proliferator-activated receptor-α (PPAR-α) signaling pathway. 16S rDNA sequencing revealed that matrine altered the composition and functions of gut microbiota, increased the abundance of Barnesiella intestinihominis and decreased the abundance of Helicobacter ganmani at the species level. In conclusion, matrine ameliorated DSS-induced colitis by improving gut barrier integrity, inhibiting the PPAR-α signaling pathway, and modulating gut microbiota. These suggested that matrine may be a therapeutic agent for UC treatment.

Role of PRPS2 as a prognostic and therapeutic target in osteosarcoma.

AIMS: Osteosarcoma (OS) is the most common primary malignant tumour of the bone. However, further improvement in survival has not been achieved due to a lack of well-validated prognostic markers and more effective therapeutic agents. Recently, the c-Myc-phosphoribosyl pyrophosphate synthetase 2 (PRPS2) pathway has been shown to promote nucleic acid metabolism and cancer cell proliferation in malignant melanoma; phosphorylated mammalian target of rapamycin (p-mTOR) has been upregulated and an effective therapeutic target in OS. However, the p-mTOR-PRPS2 pathway has not been evaluated in OS. METHODS: In this study, the expression level of PRPS2, p-mTOR and marker of proliferation (MKI-67) was observed in a cohort of specimens (including 236 OS cases and 56 control samples) using immunohistochemistry, and the association between expression level and clinicopathological characteristics of patients with OS was analysed. RESULTS: PRPS2 protein level, which is related to tumour proliferation, was higher in OS cells (p=0.003) than in fibrous dysplasia, and the higher PRPS2 protein level was associated with a higher tumour recurrence (p=0.001). In addition, our statistical analysis confirmed that PRPS2 is a novel, independent prognostic indicator of OS. Finally, we found that the expression of p-mTOR was associated with the poor prognosis of patients with OS (p<0.05). CONCLUSIONS: PRPS2 is an independent prognostic marker and a potential therapeutic target for OS.

CDKL3 promotes osteosarcoma progression by activating Akt/PKB.

Osteosarcoma (OS) is a primary malignant bone neoplasm with high frequencies of tumor metastasis and recurrence. Although the Akt/PKB signaling pathway is known to play key roles in tumorigenesis, the roles of cyclin-dependent kinase-like 3 (CDKL3) in OS progression remain largely elusive. We have demonstrated the high expression levels of CDKL3 in OS human specimens and comprehensively investigated the role of CDKL3 in promoting OS progression both in vitro and in vivo. We found that CDKL3 regulates Akt activation and its downstream effects, including cell growth and autophagy. The up-regulation of CDKL3 in OS specimens appeared to be associated with Akt activation and shorter overall patient survival (P = 0.003). Our findings identify CDKL3 as a critical regulator that stimulates OS progression by enhancing Akt activation. CDKL3 represents both a biomarker for OS prognosis, and a potential therapeutic target in precision medicine by targeting CDKL3 to treat Akt hyper-activated OS.

Network pharmacology-based identification of the protective mechanisms of taraxasterol in experimental colitis.

BACKGROUND AND AIM: Taraxasterol, a pentacyclic-triterpene, has been reported to exert potent anti-inflammatory activity. However, the molecular mechanisms by which taraxasterol attenuates acute experimental colitis (AEC) remain undocumented. METHODS: A network pharmacology approach was used to identify the candidate and collective targets of taraxasterol and acute colitis, and an AEC model was established by oral administration of dextran sulfate sodium (DSS) in mice. Body weight and colon lengths were then examined, the pathological scoring was assessed by using hematoxylin and eosin staining, and the expression levels of target genes were further confirmed by qRT-PCR and immunohistochemistry (IHC) analysis in taraxasterol treated AEC models. RESULTS: 14 collective targets of taraxasterol and acute colitis were identified by a network pharmacology analysis, including PPARG, JAK2, MMP3, NR1I2 and PTPN11. Further investigations in an AEC model showed that, taraxasterol alleviated the unfavorable clinical symptoms and attenuated the intestinal inflammation response by reducing the cytokines TNF-α, IL-1ß and IL-6 levels. qRT-PCR and IHC analysis evidenced that, taraxasterol decreased MMP3 expression levels, but increased PPARG expression levels in AEC models as compared with the DSS group. CONCLUSIONS: Our findings demonstrated that taraxasterol improved DSS-induced AEC through regulating MMP3 and PPARG expression, providing a new insight into the potential therapeutic strategies for acute colitis.

Minichromosome maintenance protein 2 and 3 promote osteosarcoma progression via DHX9 and predict poor patient prognosis.

A label free quantitative proteomic approach (SWATH™ experiment) was performed to identify tumor-associated nuclear proteins that are differentially expressed between osteosarcoma cells and osteoblast cells. By functional screening, minichromosome maintenance protein 2 (MCM2) and minichromosome maintenance protein 3 (MCM3) were found to be related to osteosarcoma cell growth. Here, we show that knockdown of MCM2 or MCM3 inhibits osteosarcoma growth in vitro and in vivo. In co-immunoprecipitation and co-localization experiments, MCM2 and MCM3 were found to interact with DExH-box helicase 9 (DHX9) in osteosarcoma cells. A rescue study showed that the decreased growth of osteosarcoma cells by MCM2 or MCM3 knockdown was reversed by DHX9 overexpression, indicating that MCM2 and MCM3 activity was DHX9-dependent. In addition, the depletion of DHX9 hindered osteosarcoma cell proliferation. Notably, MCM2 and MCM3 expression levels were positively correlated with the DHX9 expression level in tumor samples and were associated with a poor prognosis in patients with osteosarcoma. Taken together, these results suggest that the MCM2/MCM3-DHX9 axis has an important role in osteosarcoma progression.

Insights into Trx1, TRP14, and Prx1 homologs of Paralichthys olivaceus: molecular profiles and transcriptional responses to immune stimulations.

Thioredoxin (Trx) proteins are involved in several cellular processes, such as anti-oxidative stress and cellular redox homeostasis. In this study, we isolated the full-length cDNAs of PoTrx1 and PoTRP14 from Japanese flounder (Paralichthys olivaceus). PoTrx1 is 723 bp in length, with a 366-bp open reading frame (ORF) that encodes for 121 amino acids. PoTRP14 is 909 bp in length, with a 372-bp ORF that encodes for 123 amino acids. PoTrx1 and PoTRP14 are highly conserved in Cys-Gly-Pro-Cys and Cys-Pro-Asp-Cys forms, respectively. Tissue distribution analysis revealed that the transcripts of PoTrx1 and PoTRP14 were ubiquitously expressed in all tested tissues and particularly abundant in immunity-related organs, such as the liver, intestine, gill, and spleen. Development expression profiles indicated that PoTrx1 transcript was expressed from the neurula stage to the 1 day post-hatching stage; the maximum transcript levels were recorded at the somatic stage. The mRNA level of PoTRP14 was constantly expressed at all examined developmental stages, reaching the peak at the before-hatching stage. Prx1 is a peroxiredoxin family member that serves similar functions to PoTrx1 and PoTRP14. A primary hepatocyte culture system was established to examine the immunoregulatory properties of PoTrx1, PoTRP14, and Prx1 in response to lipopolysaccharide, CuSO4, and H2O2 stimulation. Results revealed that the transcript levels of PoTrx1, PoTRP14, and Prx1 were significantly up-regulated in a time-dependent manner after the immunostimulant challenge. These data suggest that PoTrx1, PoTRP14, and Prx1 play critical roles in anti-oxidation and immunoregulation.

Mixed epithelial and stromal tumor of the kidney: report of a rare case and review of literature.

Mixed epithelial and stromal tumor of the kidney (MESTK) is a rare benign tumor composed of epithelial and stromal cells. We report a rare male case with detailed clinicopathological data and follow-up information. The patient presented with gross hematuria. Computed tomography (CT) and magnetic resonance imaging study showed a 60 mm×40 mm cystic lesion with thickened septa and minimal contrast enhancement at the lower pole of the right kidney. The patient underwent nephron sparing surgery (NSS). Intraoperative frozen section showed benign histological features without significant cytologic atypia and mitosis. By additional immunohistochemistry investigations, the epithelial component was positive for cytokeratin-7, high molecular weight cytokeratin, and PAX-8. The stromal component showed strong positivity for vimentin and smooth-muscle actin. This case emphasizes that it is difficult to establish a precise diagnosis of MESTK preoperatively due to lack of any typical radiological features. Thus, intraoperative frozen section is of great clinical significance for NSS with preservation of kidney function. Additionally, regular follow-up is necessary for the MESTK with malignant potential.

[Accuracy of different preoperative biopsy techniques in diagnosis of osteosarcomas and their value in prognostic evaluation].

OBJECTIVE: To study the difference in pathologic diagnostic accuracy among different histologic subtypes of osteosarcoma and different methods of preoperative biopsy, and the influence of diagnostic accuracy on prognosis of osteosarcoma. METHODS: The preoperative biopsies, complete clinical, radiological and pathological data of 347 pathologically confirmed osteosarcomas were evaluated. According to the Pathological Diagnostic and Technical Specifications, the accuracy of preoperative biopsies was divided into 6 grades. 1: definite diagnosis, 2: basically definite diagnosis, 3: significant diagnosis, 4: descriptive diagnosis, 5:inadequate sampling, 6:misdiagnosis. 1 to 3 were defined as successful diagnosis,while 4 to 6 were defined as unsuccessful diagnosis. RESULTS: Of the 347 biopsies, 252 were CT-guided needle biopsies by the radiologists, and 95 were core-needle biopsies by orthopedic surgeons without CT-guidance. The latter showed a higher overall biopsy success rate (97.9%) in all osteosarcomas. Biopsies by surgeons showed a higher biopsy success rate (95.4%) in conventional osteosarcoma, but lower success rate in telangiectatic (55.6%) and low-grade central osteosarcomas (63.7%). The accuracy of pathologic diagnosis of preoperative biopsy was related to patients' age, serum AKP level, imaging diagnosis, method of biopsy and the subtype of osteosarcoma. Comparing the groups with successful and unsuccessful diagnosis, there were significant differences in recurrence rate and mortality after operation (P<0.01). CONCLUSIONS: The accuracy of pathologic diagnosis of preoperative biopsy are related to recurrence rate and mortality after operation. Biopsy by orthopedic surgeons without CT-guidance is reliable and safe, followed by primary diagnosis at frozen section and final diagnosis by routine pathologic sections for osteosarcomas located in the long bones of the extremities. Close integration of the preoperative pathologic diagnosis with clinical and radiological data will improve the accuracy of diagnosis.