Gut Dysbiosis Drives Inflammatory Bowel Disease Through the CCL4L2-VSIR Axis in Glycogen Storage Disease.

Patients with glycogen storage disease type Ib (GSD-Ib) frequently have inflammatory bowel disease (IBD). however, the underlying etiology remains unclear. Herein, this study finds that digestive symptoms are commonly observed in patients with GSD-Ib, presenting as single or multiple scattered deep round ulcers, inflammatory pseudo-polyps, obstructions, and strictures, which differ substantially from those in typical IBD. Distinct microbiota profiling and single-cell clustering of colonic mucosae in patients with GSD are conducted. Heterogeneous oral pathogenic enteric outgrowth induced by GSD is a potent inducer of gut microbiota immaturity and colonic macrophage accumulation. Specifically, a unique population of macrophages with high CCL4L2 expression is identified in response to pathogenic bacteria in the intestine. Hyper-activation of the CCL4L2-VSIR axis leads to increased expression of AGR2 and ZG16 in epithelial cells, which mediates the unique progression of IBD in GSD-Ib. Collectively, the microbiota-driven pathomechanism of IBD is demonstrated in GSD-Ib and revealed the active role of the CCL4L2-VSIR axis in the interaction between the microbiota and colonic mucosal immunity. Thus, targeting gut dysbiosis and/or the CCL4L2-VISR axis may represent a potential therapy for GSD-associated IBD.

Polyphenol Extracts from Ziziphus jujuba Mill. "Junzao" Attenuates Ulcerative Colitis by Inhibiting the NLRP3 and MAPKs Signaling Pathways and Regulating Gut Microbiota Homeostasis in Mice.

SCOPE: Polyphenols are the major active substances in red jujube fruit, and their anti-inflammatory and antioxidant activities suggest their potential utility in the prevention of ulcerative colitis (UC). METHODS AND RESULTS: In this study, the effect of polyphenol extracts from red jujube (Ziziphus jujuba Mill. "Junzao") (PERJ) on the dextron sulfate sodium (DSS)-induced UC mice is investigated. The result shows that PERJ effectively improves clinical symptoms, including food and water intake, the disease activity insex (DAI) and spleen index, and routine blood levels, and alleviates the shortening of the colon, in mice with DSS-induced UC. Meanwhile, PERJ remarkably decreases the expression of proinflammatory factors. Moreover, PERJ repairs intestinal barrier damage by increasing the expression level of mucin 2 and mucin 3, and the result is also confirmed in the histological assessment. Besides, the expression levels of Nod-like receptor family pyrin domain-containing 3 (NLRP3) and mitogen-activated protein kinase cascade (MAPKs) signaling pathway-related proteins are inhibited by the PERJ administration. Finally, 16S rRNA sequencing analyses reveal that PERJ reverses intestinal microbiota dysbiosis by enhancing the abundance of Firmicutes and decreasing that of Proteobacteria and Bacteroidetes. CONCLUSION: PERJ probably inhibits the development of UC by suppressing the NLRP3 and MAPKs signaling pathways and regulating gut microbiota homeostasis, and can be considered as a potential resource for preventing UC.

Silencing PKM2 Attenuates Brain Injury Induced by Status Epilepticus by Inhibiting the AKT/mTOR Pathway and the NLRP3 Inflammasome.

PKM2 is a glycolytic pyruvate kinase isoenzyme, and its role in neurological diseases has been published. However, the role and mechanism of PKM2 in the process of status epilepticus have not been reported. The purpose of this study is to explore the role and mechanism of PKM2 in epilepsy. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used to explore the expression of PKM2 in cells. Enzyme-linked immunosorbent assay kits were used to evaluate the level of inflammatory factors. An epilepsy model was established by intraperitoneal injection of lithium chloride in rats. Various behavioural assays were conducted to explore the learning ability and cognitive level of rats. PKM2 expression was upregulated in Mg2+-induced hippocampal neurons. PKM2 inhibition ameliorated Mg2+-induced hippocampal neuronal inflammation and reduced neuronal apoptosis. In addition, PKM2 silencing inhibited the metabolic dysfunction of Mg2+-induced hippocampal neurons. Subsequent experiments showed that the Akt/mTOR pathway and NLRP3 inflammasome are involved in PKM2-mediated neuronal regulation. More importantly, PKM2 inhibition could alleviate status epilepticus in rats. PKM2 inhibition attenuates Mg2+-induced hippocampal neuronal inflammation, apoptosis and metabolic dysfunction and improves the cognitive ability of rats. Therefore, PKM2 may be an important target for epilepsy treatment.

Genome-Wide Identification and Analysis of the Plant Cysteine Oxidase (PCO) Gene Family in Brassica napus and Its Role in Abiotic Stress Response.

Plant Cysteine Oxidase (PCO) is a plant O2-sensing enzyme catalyzing the oxidation of cysteine to Cys-sulfinic acid at the N-termini of target proteins. To better understand the Brassica napus PCO gene family, PCO genes in B. napus and related species were analyzed. In this study, 20, 7 and 8 PCO genes were identified in Brassica napus, Brassica rapa and Brassica oleracea, respectively. According to phylogenetic analysis, the PCOs were divided into five groups: PCO1, PCO2, PCO3, PCO4 and PCO5. Gene organization and motif distribution analysis suggested that the PCO gene family was relatively conserved during evolution. According to the public expression data, PCO genes were expressed in different tissues at different developmental stages. Moreover, qRT-PCR data showed that most of the Bna/Bra/BoPCO5 members were expressed in leaves, roots, flowers and siliques, suggesting an important role in both vegetative and reproductive development. Expression of BnaPCO was induced by various abiotic stress, especially waterlogging stress, which was consistent with the result of cis-element analysis. In this study, the PCO gene family of Brassicaceae was analyzed for the first time, which contributes to a comprehensive understanding of the origin and evolution of PCO genes in Brassicaceae and the function of BnaPCO in abiotic stress responses.

Drug therapy for bone metastasis of malignant tumor: theory, progress, and potential.

Bone is a common metastatic site of malignancies, caused by the complex interaction between tumor cells and the bone microenvironment. The complicated procedure covers multiple targets for therapeutic strategies against bone metastasis. At the present, only bisphosphonates and denosumab are currently approved for the prevention of skeletal-related events. But it is still ineffective for some patients, and none of them are proven to prolong the overall survival of patients with bone metastasis. Thus, new bone-modifying agents and therapeutic strategies are required. The review aimed to generalize the basic theory of bone metastasis and major put emphasis on the development of fundamental and potential target drugs in the behavior of bone metastasis. The summary of the drug development process helps to provide ideas for finding new and effective treatments for bone metastasis.

Clinical and laboratory parameters predicting cancer in dermatomyositis patients with anti-TIF1γ antibodies.

BACKGROUND: Dermatomyositis (DM) is a chronic acquired autoimmune disorder strongly associated with cancer development. Until now, identifying predictive markers indicating a high risk of cancer has challenged clinicians. Although anti-TIF1γ antibody is a major serological indicator for cancer-associated DM, many anti-TIF1γ antibody-positive DM patients lack malignancy. OBJECTIVES: To determine clinical and laboratory parameters that support cancer prediction in anti-TIF1γ antibody-positive DM patients. METHODS: Clinical and laboratory data were collected from cancer-associated and unassociated DM patients with anti-TIF1γ antibodies. Serum cytokine concentrations were measured with a cytokine array assay. The values of inflammatory cytokines in cancer prognosis were determined with a receiver operating characteristic curve analysis. RESULTS: The cancer group had a significantly higher frequency of males, older mean age and higher anti-TIF1γ antibody levels. Some inflammatory cytokines, particularly tumour necrosis factor (TNF) and TNF receptor superfamilies, had increased levels in sera that were correlated with myositis markers, cutaneous severity and DM disease activity. Moreover, these cytokines had an area under the curve (AUC) ≥0.8 and high sensitivity and specificity at their specific cut-off, even higher than anti-TIF1γ levels in cancer prediction in our DM patients. CONCLUSIONS: Our results suggest a close pathophysiological relationship among myositis, cancer and skin involvements in DM patients with anti-TIF1γ antibodies and the potential clinical significance of anti-TIF1γ antibody levels in evaluating disease severity and prognosis in DM patients. Some inflammatory cytokines, particularly TNF and TNF receptor superfamilies including BAFF, sTNF-R1 and sTNF-R2, may support cancer prediction in DM patients with anti-TIF1γ antibodies.

Development of a novel drug targeting delivery system for cervical cancer therapy.

'Targeting peptides' have demonstrated their value in diagnostic imaging and therapy and novel peptide probes specific to cervical cancer were developed. In the M13KE phage dodecapeptide (12-mer) peptide library, the phage clone S7 showed the best binding to the cancer cells as confirmed by immunofluorescence and flow cytometry assays, and was selected for continued studies. Its binding peptide, CSP3, was synthesized from the sequence of S7's 12-mer at the N-terminus of the minor coat protein pIII of this M13KE phage vector. The peptide's binding was analyzed by the same assays used for S7. It was also assessed using competitive inhibition and binding to a tissue chip. The results demonstrated that the CSP3 peptide bound to cervical carcinoma cells with high sensitivity and specificity. The positive results indicated that the peptide CSP3, conjugated with nanomaterials and chemotherapeutics, may be developed as a targeting vehicle for therapeutic drug delivery against cervical cancer, especially cervical cancer with multiple drug resistance. For this aim, we prepared a CSP3 conjugated liposome drug delivery system containing doxorubicin (DOX) and microRNA101 (miR101) expression plasmids (CSP3-Lipo-DOX-miR101), and the primary result showed that the system demonstrated significantly enhanced cytotoxicity to SiHa cells and DOX resistant SiHa cells, SiHa/ADR. Our results showed that CSP3 is a cervical cancer targeting 12aa peptide with high specificity and sensitivity, and the CSP3 conjugated drug delivery system, CSP3-Lipo-DOX-miR101 has promising potential for development as an efficient drug system for the therapy of cervical cancer.

[Methyltransferase inhibitor BIX01294 promotes the migration and inhibits decidualization of mouse uterine stromal cells in vitro].

OBJECTIVE: To investigate the effect of BIX01294 (BIX), a methyltransferase inhibitor, on the migration and decidualization of the stromal cells in mouse uterus. METHODS: Mouse endometrial stromal cells were isolated and cultured from the uterus of pregnant mice on day 3.5 of gestation. The migration and decidualization of mouse endometrial stromal cells treated with BIX at different concentrations were observed with wound healing assay and real-time PCR. RESULTS: The migration distance of mouse endometrial stromal cells increased as the BIX concentration increased within the range below 15 µmol/L. Compared with the control cells, the cells treated with BIX (15 µmol/L) showed significantly increased migration distances, but increasing BIX concentration to 20 µmol/L did not further increase the cell migration distance and began to cause cell death. Compared with the control cells, the BIX-treated stromal cells exhibited significantly down-regulated expression of Ehmt2 mRNA, and 15 µmol/L BIX caused inhibition of decidualization in the stromal cells. CONCLUSION: Within a defined concentration range, BIX promotes the migration and inhibits decidualization of mouse uterine stromal cells by inhibiting the expression of Ehmt2 mRNA.

Screening and identification of a specific peptide binding to cervical cancer cells from a phage-displayed peptide library.

OBJECTIVES: To screen and identify the probe markers specifically binding to human cervical cancer, a phage-displayed 12-mer peptide library was used for biopanning of SiHa cells. RESULTS: After four rounds of whole-cell subtraction biopanning, the phage recovery was 21-fold higher (from 3.9 × 10-5 to 8.3 × 10-4) than that of the first round, and specific phage clones were significantly enriched. 57 randomly selected phage clones were tested by ELISA, and 36 phage clones were identified as positive clones. After sequencing of positive clones, six different peptide sequences were obtained and CSP3 showed best affinity and specificity to SiHa cells via immunofluorescence assay. CONCLUSIONS: Peptide, CSP3, bound to SiHa cells specifically and sensitively. It may be a potential candidate for molecular imaging detection and targeting therapy of cervical cancer.

[Expression of FABP7 in mouse placenta tissue and human trophoblast HTR-8/Svneo cells].

OBJECTIVE: To detect the expression of FABP7 in the placenta of pregnant mice and in HTR-8/Svneo cells. METHODS: Real-time PCR and immunofluorescence were used to detect FABP7 mRNA and protein expressions in the uterine and placental tissue of pregnant mice at different days of gestation. FABP7 expression was also detected in cultured HTR-8/Svneo cells using immunofluorescence assay. The mice were treated with E2, P4 or their combination for 6 and 24 h and Fabp7 mRNA level in the uterus was detected with real-time PCR. RESULTS: At 7.5-10.5 days of gestation, the pregnant mice showed positive expressions of Fabp7 mRNA in the uterus and placenta, and FABP7 protein was detected in the decidualized cells and trophoblast giant cells. The expressions of FABP7 were detected at both the mRNA and protein levels in cultured HTR-8/Svneo cells. In mice treated with P4 alone or with E2+P4 for 6 and 24 h, the expression level of Fabp7 mRNA was upregulated in the uterus. Fabp7 upregulation was observed in mice at 24 h following E2 treatment but not at 6 h. CONCLUSION: FABP7 is expressed in trophoblast giant cells and decidual cells in the placental tissue of mice and in cultured HTR-8/Svneo cells, suggesting the involvement of FABP7 in placental development and in maintenance of pregnancy. E2 and P4 can regulate the expression of FABP7 in mouse uterus.