IL-17 Imbalance Promotes thePyroptosis in Immune-Mediated L

Autoimmune hepatitis (AIH) affects all age group and occurs mainly in women. Pyroptosis is a novel programmed cell death featured with cell bursting and release of proinflammatory cytokines. A deeper understanding of AIH pathogenesis will contribute to novel therapy for AIH patients. Here, we aimed to investigate the role of IL-17 in immune-mediated liver injury.The levels of cytokines were measured by ELISA, and mRNA levels of STAT3 and IFN gammainducible protein 16 (IFI16) were detected by PCR. Expressions of STAT3, IFI16, gasdermin D and cleaved caspase-1 were measured by western-blotting. Immunohistochemical staining and transmission electron microscopy were applied to evaluate liver histopathological changes of the treated mice. Our results showed that the levels of IFI16 was increased in hepatocytes treated with IL-17 protein, and further elevated after STAT3-overexpressed (STAT3-OE) lentivirus treatment. The levels of IFI16 were reduced in hepatocytes treated with IL-17 neutralizing Ab (nAb), but were significantly increased after STAT3-OE treatment. Pyroptosis was observed in hepatocytes treated with IL-17 protein, and further cell damage was observed after STAT3-OE lentivirus treatment. Liver damage was alleviated in mice treated with IL-17 nAb, however sever damage was experienced after STAT3-OE lentivirus treatment. A binding interaction between IFI16 and STAT3 was detected in IL-17 treated hepatocytes. Glutathione transaminase activity was enhanced in concanavalin A-induced AIH mice compared to the control group (p<0.01). IL-17 plays an important role in activating STAT3 and up-regulating IFI16, which may promote the pyroptosis in AIH-related liver injury through STAT3-IFI16 axis.

DLL3 promotes progression of prostate cancer through MAPK pathway / 中国病理生理杂志

AIM:To explore the impact of Delta-like ligand 3(DLL3)on the progression of prostate cancer(Pca)and to elucidate its potential molecular mechanisms.METHODS:Overexpression and interference plasmids of DLL3 gene were constructed,and the effects of DLL3 on the proliferation,migration and invasion of Pca cells were as-sessed.Tumorigenesis assay was employed to determine whether DLL3 influenced the proliferation of PC-3 cells in vivo.The impact of DLL3 on epithelial-mesenchymal transition(EMT)and mitogen-activated protein kinase(MAPK)signaling was investigated through Western blotting.RESULTS:Overexpression of DLL3 significantly enhanced the proliferation,migration,invasion and EMT processes of Pca cells compared with empty vector group and blank control group(P＜0.05).Conversely,knockdown of DLL3 expression yielded opposite effects(P＜0.05).Moreover,up-regulation of DLL3 promoted tumor growth in the nude mouse xenograft model(P＜0.05).Further investigation demonstrated that DLL3 up-regulation led to increases in the levels of MAPK signaling pathway-related proteins.Interestingly,MAPK inhibitor effec-tively reduced the proliferation,migration and invasion of Pca cells caused by DLL3 overexpression(P＜0.05).CON-CLUSION:DLL3 promotes the proliferation,migration and invasion of Pca through the MAPK pathway.

A four-protein metabolon assembled by a small peptide protein creates the pentacyclic carbonate ring of aldgamycins

Organic carbonates (OCs) are a class of compounds featured by a carbonyl flanked by two alkoxy/aryloxy groups. They exist in either linear or cyclic forms, of which the majority encountered in nature adopt a pentacyclic structure. However, the enzymatic basis for pentacyclic carbonate ring formation remains elusive. Here, we reported that a four-protein metabolon (AlmUII-UV) assembled by a small peptide protein (AlmUV) appends a reactive

Application of circulating tumor DNA in diagnosis and prognosis of epithelial ovarian cancer / 中华全科医师杂志

In the development of cancer, the DNA fragments of tumor cells enter into the blood circulation to form circulating tumor DNA (ctDNA), which is an indicator for detecting and quantifying tumor mutations. Epithelial ovarian cancer accounts for most of the histological types in ovarian cancer. This article reviews the clinical application of ctDNA in epithelial ovarian cancer, including the early diagnosis, therapeutic effect evaluation after operation and adjuvant therapy, tumor load assessment, dynamic monitoring during the follow-up and the prognosis of patients.

Construction and expression of the fusion gene CCK39/UreB in recombinant Escherichia coli BL-21(DE3) / 生物工程学报

The aims of this research were to construct prokaryotic expression vector containing fusion gene of Cholecystokinin 39 (CCK39) of pig and Urease subunit B (UreB) of coliform bacteria, and then to express the fusion protein in recombinant Escherichia coli BL21(DE3). The CCK39 gene was amplified by RT-PCR from the extracted total RNA of pig's duodenum, and the UreB gene was then amplified by PCR from the extracted plasmid DNA of bacillus of coliform bacteria from pig's intestinal content. Then the CCK39 and the UreB were inserted into the prokaryotic expression vector pET43a(+) to construct a recombinant fusion expression vector pET43a(+)/CCK39/UreB and then, the recombinant vector was identified by PCR, endonuclease digestion and sequence analysis. It was identified that the gene fragment of CCK39 at length of 117 bp and UreB at length of 324 bp were amplified and cloned into the vector pET43a(+) successfully. The recombinant vector was transformed into Escherichia coli BL21(DE3) and induced the expression of CCK39/UreB fusion protein with a molecular mass of approximately 80 kD by using isopropylthio-beta-D-galactoside (IPTG) as inducer. The fusion protein was mostly located in the cytoplasm and it was soluble. The soluble protein was collected and purified by Ni2+-NTA column chromatograph and then reached a purity of more than 95%. It was proved by western blotting that the fusion protein could react with rabbit anti-CCK8 antiserum and rabbit anti-UreB antiserum. Therefore, the expressed fusion protein has good antigenicity. This work established a good foundation for further study on the production of anti-CCK/Urease vaccines.