Glutathione-S-transferase A3 protein suppresses thiram-induced tibial dyschondroplasia by regulating prostaglandin-related genes expression.

Tibial dyschondroplasia (TD) is an intractable avian cartilage disease in which proximal growth plates of tibia lack blood vessels and contain nonviable cells, and it leads to the inflammatory response. Prostaglandins (PGs) genes have not been studied yet in TD chicken, and they might play role in skeletal metabolism, therefore we planned to explore the role of recombinant glutathione-S-transferase A3 (rGSTA3) protein and PG-related genes. In this study, qRT-PCR, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC) analysis were used to identify the expression patterns of eight PG-related genes in the tibial growth plate of broiler chicken. The results showed that the expression of PG-related genes glutathione-S-transferase A3 (GSTA3), cyclooxygenase 2 (COX-2), prostaglandin D2 synthase (PTGDS), prostaglandin E synthase (PTGES), prostaglandin E2 receptor (PTGER) 3, PTGER4, prostaglandin reductase 1 (PTGR1) and hematopoietic prostaglandin D synthases (HPGDS) expression were identified and could significantly respond to thiram-induced TD chicken. Interestingly, the expression of rate-limiting enzyme COX-2 and PGE2 were induced after the treatment of rGSTA3 protein. These findings demonstrated that the occurrence of TD is closely related to the inhibition of PGs. Moreover, rGSTA3 protein participated in the recovery of TD by strengthening the expression of PG-related genes.

MicroRNA-26a suppresses the malignant biological behaviors of papillary thyroid carcinoma by targeting ROCK1 and regulating PI3K/AKT signaling pathway.

OBJECTIVE: Papillary thyroid carcinoma (PTC) is an endocrine malignancy, the morbidity of which has kept rising in recent years. MicroRNA (miRNA/miR) is emerging as a key regulator in carcinogenesis, including PTC. The current study concentrates on the biological roles and mechanisms of miR-26a in the PTC progression. PATIENTS AND METHODS: 51 pairs of PTC tissue samples and matched adjacent thyroid tissues were collected from PTC patients who received surgical excisions at The People's Hospital of Linqing between July 2015 and June 2018 with informed consent. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to detect expressions of miR-26a and rho-associated coiled-coil-containing protein kinase 1 (ROCK1) mRNA in PTC tissues and cells. Functional assays, including (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) MTT assays and transwell assays were performed to determine the roles of miR-26a in the PTC progression. Western blot was used to detect expression levels of the related proteins. RESULTS: Findings demonstrated prominently down-regulated miR-26a in PTC tissues and cells. Down-regulated miR-26a indicated malignant clinicopathologic characteristics and shorter overall survival rate of PTC patients. MTT assay and transwell assay indicated that miR-26a up-regulation significantly repressed PTC cell viability, invasion, and metastasis. Western blot analysis revealed that miR-26a exerted its anti-PTC effects via phosphatidylinositol 3-kinase/protein kinase B pathway (PI3K/AKT) and epithelial-to-mesenchymal transition (EMT). ROCK1 was a target of miR-26a in PTC cells and ROCK1 was mediated by miR-26a, as a regulatory mechanism in PTC. CONCLUSIONS: Taken together, these findings demonstrated the anti-tumor functions of miR-26a in PTC, providing novel strategies for PTC diagnosis and therapy.

Role of death receptors in the regulation of hepatocyte proliferation and apoptosis during rat liver regeneration.

The balance between hepatocyte proliferation and apoptosis is critical for liver homeostasis during liver regeneration. We created a rat liver regeneration model by partial hepatectomy (PH) to investigate the overall mechanism that regulates the proliferation and apoptosis of hepatocytes. The Rat Genome 230 2.0 Array was used to investigate changes in the expression levels of genes associated with the known proliferation or apoptosis signaling pathways. Ingenuity Pathway Analysis 9.0 was used to determine interactions among these signaling pathways. The results revealed that the expression levels of multiple key genes in three death receptor (DR) pathways, Fas/FasL, TNFR/TNFα, and DR6, were significantly altered in hepatocytes after PH. The expression level of the gene encoding DR6 increased by over 100-fold, whereas the levels of the genes encoding Fas, FasL, and TNFα were increased by 2-4-fold 12 h after PH. Fas/FasL, TNFR/TNFα, and DR6 are known to participate in numerous cellular events including cell proliferation and apoptosis. Our results suggest that the DR6 pathway plays a major role in the regulation of hepatocyte apoptosis, whereas Fas/FasL and TNFR/TNFα pathways may have roles in coordinating signaling activities between proliferation and apoptosis.

Effect of nimodipine on rat spinal cord injury.

We evaluated the potentially protective effect of nimodipine on rat spinal cord injury. Sprague-Dawley rats received spinal cord injury, and were separated into nimodipine (N = 12) and saline groups (N = 12). Within 1 h of the injury, rats were treated intraperitoneally with nimodipine (1.0 mg/kg) or an equal amount of saline. Treatment was performed 3 times a day for 1 week. Operation BBB score and track experiment were used to measure the physical function of the hind legs 1 and 2 weeks after injury. Two weeks after the injury, malondialdehyde (MDA) content and spinal cord myeloperoxidase (MPO) activity of the injured part were determined, and the glial scar and dead room were studied using the immune tissue chemical test. ED1 was used to observe active gitter cell and macrophages. The physical function of the nimodipine group improved significantly (P < 0.01). Two weeks after injury, spinal cord MDA content in the spinal cord in the nimodipine group (nmol/g, 25.6 ± 9.7 vs 68.5 ± 16.7) and MPO activity (U/g, 252.2 ± 63.9 vs 382.8 ± 108.2) decreased significantly (P < 0.01); nimodipine whole dead space (mm2, 4.45 ± 1.28 vs 6.16 ± 2.65) and ED1 antibody immunity colored positive room (mm2, 1.87 ± 0.42 vs 2.86 ± 1.01) reduced significantly (P < 0.01). Nimodipine treatment could reduce oxidative injury after spinal cord injury, reduce the whole dead space and inflammation, and repair spinal cord injury.

Studies on different iron source absorption by in situ ligated intestinal loops of broilers.

The objective of this study was to investigate the iron source absorption in the small intestine of broiler. In situ ligated intestinal loops of 70 birds were poured into one of seven solutions, including inorganic iron (FeSO4, Fe2(SO4)3), organic Fe glycine chelate (Fe-Gly(II), Fe-Gly(III)), the mixtures (FeSO4 with glycine (Fe+Gly(II)), Fe2(SO4)3 with glycine (Fe+Gly(III)), and no Fe source (control). The total volume of 3-mL solution (containing 1 mg of elemental Fe) was injected into intestinal loops, and then 120-min incubation was performed. Compared with inorganic iron groups, in which higher FeSO4 absorption than Fe2(SO4)3 was observed, supplementation with organic Fe glycine chelate significantly increased the Fe concentration in the duodenum and jejunum (P < 0.05), however, decreased DMT1 and DcytB messenger RNA (mRNA) levels (P < 0.05). Organic Fe glycine chelate (Fe-Gly(II), Fe-Gly(III)) increased serum iron concentration (SI), compared with inorganic 3 valence iron groups (Fe2(SO4)3 and Fe+Gly(III)) (P < 0.05); moreover, lower TIBC value was observed for the chelate (P < 0.05); however, mixture of inorganic iron and glycine did not have a positive role at DMT1 and DcytB mRNA levels, SI and Fe concentrations in the small intestine. Those results indicated that the absorption of organic Fe glycine chelate was more effective than that of inorganic Fe, and the orders of iron absorption in the small intestine were: Fe-Gly(II), Fe-Gly(III) > FeSO4, Fe+Gly(II) > Fe2(SO4)3, Fe+Gly(III). Additionally, the simple mixture of inorganic iron and glycine could not increase Fe absorption, and the duodenum was the main site of Fe absorption in the intestines of broilers and the ileum absorbed iron rarely.

Long-term survival of non-small-cell lung cancer patients with EGFR inhibitor treatment.

The epidermal growth factor receptor (EGFR) inhibitors gefitinib and erlotinib are effective in the treatment of advanced non-small-cell lung cancer (NSCLC), but the median survival of patients is short. Here, we describe 2 patients with NSCLC receiving conventional chemotherapy and alternative treatment with gefitinib or erlotinib as second-line therapy. The first patient was alive at 8 years with alternative conventional chemotherapy and gefitinib, and the second patient was alive at long-term follow-up with conventional chemotherapy and gefitinib or erlotinib. Gefitinib, erlotinib, and conventional chemotherapy can be combined for satisfactory therapy for NSCLC.

Cyclosporine induces up-regulation of immunoglobulin-like transcripts 3 and 4 expression on and activity of NKL cells.

BACKGROUND: Immunoglobulin-like transcripts (ILTs), which belong to a kind of receptor family discovered recently, are differentially expressed on myeloid and lymphoid cells. Most of them play important roles to regulate human immune responses by interacting with ligands. Cyclosporine (CsA) is frequently used to prevent graft-versus-host disease and treat autoimmune diseases. There are some studies about the effects of CsA on various human immunologic reactions, but its impact on ILT3 and ILT4 expression on natural killer (NK) cells is less well understood. METHODS: An NKL cell line was exposed to CsA (5, 10, 15, or 20 mg/L) for 12, 24, or 36 hours before real-time quantitative polymerase chain reaction and flow cytometry were used to detect alterations in ILT3 and ILT4 mRNA and protein expressions. NKL cells treated for 36 hours with or without CsA (15 mg/L) and then coincubated with BGC-823 or JEG-3 cells, in cytolytic and proliferative systems measured by Thiazoyl blue tetrazolium bromide assays. RESULTS: After CsA treatment both RNA and protein levels of ILT3 and ILT4 on NKL cells were increased for 12, 24, or 36 hours. CsA at various concentrations inhibited the proliferation of NKL cells to varying degrees; at 36 hours CsA (15 mg/L) showed greater effects on ILT3 and ILT4 expression and less influence on NKL growth. The ability of NKL cells primed with CsA (15 mg/L) for 36 hours to kill tumor cells was decreased markedly. CONCLUSIONS: CsA up-regulated the expression of ILT3 and ILT4 on NKL cells, which influenced their cytotoxicity against tumor cells with different expression of HLA-G and proliferation of NKL cells.