Genome-wide QTL mapping for stripe rust resistance in spring wheat line PI 660122 using the Wheat 15K SNP array.

Introduction: Stripe rust is a global disease of wheat. Identification of new resistance genes is key to developing and growing resistant varieties for control of the disease. Wheat line PI 660122 has exhibited a high level of stripe rust resistance for over a decade. However, the genetics of stripe rust resistance in this line has not been studied. A set of 239 recombinant inbred lines (RILs) was developed from a cross between PI 660122 and an elite Chinese cultivar Zhengmai 9023. Methods: The RIL population was phenotyped for stripe rust response in three field environments and genotyped with the Wheat 15K single-nucleotide polymorphism (SNP) array. Results: A total of nine quantitative trait loci (QTLs) for stripe rust resistance were mapped to chromosomes 1B (one QTL), 2B (one QTL), 4B (two QTLs), 4D (two QTLs), 6A (one QTL), 6D (one QTL), and 7D (one QTL), of which seven QTLs were stable and designated as QYrPI660122.swust-4BS, QYrPI660122.swust-4BL, QYrPI660122.swust-4DS, QYrPI660122.swust-4DL, QYrZM9023.swust-6AS, QYrZM9023.swust-6DS, and QYrPI660122.swust-7DS. QYrPI660122.swust-4DS was a major all-stage resistance QTL explaining the highest percentage (10.67%-20.97%) of the total phenotypic variation and was mapped to a 12.15-cM interval flanked by SNP markers AX-110046962 and AX-111093894 on chromosome 4DS. Discussion: The QTL and their linked SNP markers in this study can be used in wheat breeding to improve resistance to stripe rust. In addition, 26 lines were selected based on stripe rust resistance and agronomic traits in the field for further selection and release of new cultivars.

Bavachin Induces Ferroptosis through the STAT3/P53/SLC7A11 Axis in Osteosarcoma Cells.

Ferroptosis is a new form of regulated cell death, which is mediated by intracellular iron. Although it is reported that bavachin has antitumour effects on several tumour cells and prompts the reactive oxygen species (ROS) generation, it is unclear whether ferroptosis can be induced by bavachin in osteosarcoma (OS) cells. In this study, we found that bavachin inhibits the viability of MG63 and HOS OS cell lines along with an increase in the ferrous iron level, ROS accumulation, malondialdehyde overexpression, and glutathione depletion. Moreover, iron chelators (deferoxamine), antioxidants (Vit E), and ferroptosis inhibitors (ferrostatin-1 and liproxstatin-1) reverse bavachin-induced cell death. Bavachin also altered the mitochondrial morphology of OS cells, leading to smaller mitochondria, higher density of the mitochondrial membrane, and reduced mitochondrial cristae. Further investigation showed that bavachin upregulated the expression of transferrin receptor, divalent metal transporter-1, and P53, along with downregulating the expression of ferritin light chain, ferritin heavy chain, p-STAT3 (705), SLC7A11, and glutathione peroxidase-4 in OS cells. More importantly, STAT3 overexpression, SLC7A11 overexpression, and pretreatment with pifithrin-α (P53 inhibitor) rescued OS cell ferroptosis induced by bavachin. The results show that bavachin induces ferroptosis via the STAT3/P53/SLC7A11 axis in OS cells.

Intelligent rehabilitation assistant system to promote the early functional recovery of the elderly patients with femoral neck fracture after hemiarthroplasty (HA): A protocol for a randomized controlled trial.

BACKGROUND: Femoral neck fracture is a common type of hip fracture, which has a high morbidity and mortality. Surgical treatment is the first choice. However, the functional rehabilitation after operation has not been paid enough attention. In addition, the quality of exercise is difficult to quantify, and the rehabilitation is lack of standards. Therefore, the intelligent rehabilitation assistant system which could record exercise details, might be used to evaluate the quality and adherence to the prescribed exercise to this fragile group of patients has great relevance, so as to provide new ideas for postoperative rehabilitation of hip fracture. METHODS: This is an opening, prospective, double-dummy RCT. Fifty femoral neck fractures patients, older than 65 years and are about to hospitalize for HA, will be invited to study. The sample will be divided into monitoring group and control group randomly at a 1:1 ratio. The prescribed exercises need to be done continuously for 2 weeks. The monitoring group needs additional use intelligent rehabilitation assistant system. Each subject will receive a total of 4 follow-up visits at the designated time (2 weeks, 4 weeks, 12 weeks, and 24 weeks). The following factors will be talked as dependent variables:Each subject will receive a total of 4 follow-up visits at the designated time, and the findings will be analyzed statistically considering a 5% significance level (P < .05). DISCUSSION: Exercise under monitor may improve patients compliance and exercise quality, and accelerate the rehabilitation process. This protocol reported in accordance with the CONSORT 2010 checklist and SPIRIT 2013 Checklist. TRIAL REGISTRATION: The trial is registered at Chinese Clinical Trials Registry (ChiCTR2000033213, May 24, 2020).

Lineage-specific exosomes promote the odontogenic differentiation of human dental pulp stem cells (DPSCs) through TGFß1/smads signaling pathway via transfer of microRNAs.

BACKGROUND: Exosomes derived from dental pulp stem cells (DPSCs) can be used as biomimetic tools to induce odontogenic differentiation of stem cells, but the regulatory mechanisms and functions of exosome-encapsulated microRNAs are still unknown. The present study aimed to clarify the role of microRNAs contained in the exosomes derived from human DPSCs and their potential signaling cascade in odontogenic differentiation. METHODS: Exosomes were isolated from human DPSCs cultured undergrowth and odontogenic differentiation conditions, named UN-Exo and OD-Exo, respectively. The microRNA sequencing was performed to explore the microRNA profile contained in UN-Exo and OD-Exo. Pathway analysis was taken to detect enriched pathways associated with the predicted target genes of microRNAs. The regulatory roles of a highly expressed microRNA in OD-Exo were investigated through its inhibition or overexpression (miRNA inhibitors and miRNA mimics). Automated western blot was used to identify the function of exosomal microRNA and the roles of TGFß1/smads pathway in odontogenic differentiation of DPSCs. A luciferase reporter gene assay was used to verify the direct target gene of exosomal miR-27a-5p. RESULTS: Endocytosis of OD-Exo triggered odontogenic differentiation of DPSCs by upregulating DSP, DMP-1, ALP, and RUNX2 proteins. MicroRNA sequencing showed that 28 microRNAs significantly changed in OD-Exo, of which 7 increased and 21 decreased. Pathway analysis showed genes targeted by differentially expressed microRNAs were involved in multiple signal transductions, including TGFß pathway. 16 genes targeted by 15 differentially expressed microRNAs were involved in TGFß signaling. Consistently, automated western blot found that OD-Exo activated TGFß1 pathway by upregulating TGFß1, TGFR1, p-Smad2/3, and Smad4 in DPSCs. Accordingly, once the TGFß1 signaling pathway was inhibited by SB525334, protein levels of p-Smad2/3, DSP, and DMP-1 were significantly decreased in DPSCs treated with OD-Exo. MiR-27a-5p was expressed 11 times higher in OD-Exo, while miR-27a-5p promoted odontogenic differentiation of DPSCs and significantly upregulated TGFß1, TGFR1, p-Smad2/3, and Smad4 by downregulating the inhibitory molecule LTBP1. CONCLUSIONS: The microRNA expression profiles of exosomes derived from DPSCs were identified. OD-Exo isolated under odontogenic conditions were better inducers of DPSC differentiation. Exosomal microRNAs promoted odontogenic differentiation via TGFß1/smads signaling pathway by downregulating LTBP1.

Indolent T-lymphoblastic proliferation associated with Castleman disease and low grade follicular dendric cell sarcoma: report of a case and review of literature.

Indolent T-lymphoblastic proliferation (IT-LBP) is referred to as extrathymic immature TDT+T-cell hyperplasia, non-neoplastic lesion, often misdiagnosed as T-lymphoblastic lymphoma and overtreated. We report a case of a 20-year-old male patient with a right adrenal gland mass, diagnosed as indolent T-lymphoblastic proliferation (IT-LBP) associated with hyaline vascular Castleman disease (HV-CD) and low grade follicular dendric cell sarcoma (LG-FDCS). The case is a rare combination of finding, and it is the first case occurring in adrenal gland. IT-LBP is a clinically indolent disease, requiring no treatment, often associated with other tumors. Because of the high ki67 index, IT-LBP is easily misdiagnosed as T-lymphoblastic lymphoma, causing overtreatment. Understanding the biological behavior, treatment, prognosis and the associated diseases of IT-LBP is important.

Analysis of DNA methylation patterns associated with the gastric cancer genome.

The objective of the current study was to investigate the characteristics of DNA methylation patterns associated with the gastric cancer genome and to identify clinically useful diagnostic markers and therapeutic targets for gastric cancer. The Infinium 450K methylation microarray was used to compare differential DNA methylation sites of gastric cancer tissue with that of normal gastric tissue. The results of the DNA microarray analysis were confirmed by pyrosequencing. Functional analysis of the differential genes was performed using the GO software. The effect of candidate site methylation on gene expression was monitored using quantitative polymerase chain reaction analysis. Of the 2,645 differential methylation sites identified in gastric cancer tissues, 2,016 were hypermethylated sites, 629 were hypomethylated sites, 826 were located in promoter regions and 1,024 were located within genes. These differential sites were associated with 1,352 genes. In total, five sites were selected and pyrosequencing verified the results of the microarray analysis in five of the sites. Change in gastric cancer DNA methylation pattern was a common occurrence. Differential methylation sites appeared more often in non-promoter regions. The associated genes were involved in multiple signaling pathways, and hypermethylated and hypomethylated sites were involved in roughly the same signaling pathways. Methylation of the genome promoted gene expression. TRIM15, ITGAM, MSX2 and FAM38A may be candidate genes for diagnosing gastric cancer.

Matrix metalloproteinase-9 overexpression is closely related to poor prognosis in patients with colon cancer.

BACKGROUND: Matrix metalloproteinase-9 (MMP-9) is an important member of the matrix metalloproteinase family and is considered to be involved in the invasion and metastasis of cancer cells. This study analyzed the expression of MMP-9 in colon cancer patients and the relationship between this expression and clinicopathological features and survival. METHODS: We immunohistochemically investigated 68 specimens of colon cancer tissues and corresponding distal normal mucosa tissues using MMP-9 antibody. Then, the correlation between MMP-9 expression and clinicopathological features and its prognostic relevance were determined. RESULTS: The expression rate of MMP-9 in colon cancer tissues was significantly higher than that in distal normal mucosa (69.1% versus 2.9%, P < 0.001). Significant correlations were only found between high levels of MMP-9 expression and metastasis of lymph nodes and Dukes' stage. Overexpression of MMP-9 was associated with shorter survival times in univariate analysis. Multivariate analysis confirmed that MMP-9 expression was an independent prognostic factor. CONCLUSIONS: MMP-9 is correlated with the metastasis of lymph nodes, and its elevated expression may be an adverse prognostic indicator for the patients of colon cancer.

Toll-like receptor 4/nuclear factor-kappa B pathway is involved in myocardial injury in a rat chronic stress model.

Chronic stress is considered to predispose to various cardiovascular events such as coronary artery disease, hypertension, and even heart failure. In this study, rats were exposed to stress for 1 day, 1, 2, 3, and 4 weeks to establish a chronic stress model. A specific toll-like receptor 4 (TLR4) antagonist eritoran was used to block the activity of TLR4. On the second day after the last stress exposure, the animals were killed. The expression of TLR4 mRNA and nuclear factor-kappa B (NF-κB) DNA-binding activity in the myocardium were measured using reverse transcriptase polymerase chain reaction and electrophoretic mobility shift assay. The proinflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL-6) in myocardium were assayed by enzyme-linked immunosorbent assay. Myocardial injury was evident after chronic stress for 2 weeks. The TLR4 mRNA expression reached a peak after stress for 1 week. It was sustained at a stable level after stress exposure for 3 weeks and was restored to a nearly normal level in the fourth week. NF-κB DNA-binding activity was significantly enhanced after the stress for 1 day and markedly enhanced again after a 2-week stress exposure. It was weakened and reached a normal level after stress exposure for 4 weeks. The levels of TNF-α and IL-6 gradually increased and reached peaks after stress for 4 weeks. Meanwhile, eritoran significantly decreased the TLR4 mRNA expression and NF-κB activity in rats from the 2-week stress group. However, it did not downregulate the levels of TNF-α and IL-6. Importantly, it significantly improved the myocardial injury induced by the chronic stress. In conclusion, TLR4/NF-κB participates in myocardial injury during chronic stress.

The effect of lipopolysaccharides on the expression of CD14 and TLR4 in rat Kupffer cells.

OBJECTIVE: To assess the effect of lipopolysaccharides (LPS) on the expression of CD14 and TLR4 in rat Kupffer cells (KCs). METHODS: In rat KCs induced by LPS, the changes of CD14 and TLR4 expression were measured by RT-PCR and immunohistochemistry, and the expressions of TNF-alphamRNA, IL-6mRNA or the concentrations of TNF-alpha, IL-6 were estimated by in situ hybridization, radioimmunoassay, and others. RESULTS: The expressions of CD14 and TLR4 in KCs induced by LPS were markedly increased in a dose-dependent manner (10 mg/L-1 microg/L) or in a time-dependent manner (0.5 h-24 h), with the peaked expression of CD14 at 3-6 hours. The expressions of CD14 and TLR4 in KCs stimulated by the active mediators from KCs which had been exposed to LPS for 1 hour were obviously increased. CONCLUSIONS: There is a close relationship between LPS or the active mediators from KCs induced by LPS and the expressions of CD14, TLR4. It is implied that the increase of TLR4, CD14 expression may be induced by LPS within 1-3 hours, and further increase of TLR4, CD14 expression may be correlated with the cytokines produced by KCs.

[The role of LPS in the CD14 expression and the activation of Kupffer cells].

OBJECTIVE: To explore the effect of LPS on the expression of CD14 and the activation of Kupffer cells (KCs). METHODS: Rat KCs were isolated and cultured with LPS. Immunohistochemistry and RT-PCR methods were employed to determine the changes in the CD14 expression and the concentration of TNFalpha, IL-6 and NO in the supernatant of the cultured KCs with LPS. RESULTS: (1) The expression of CD14mRNA and the synthesis of CD14 protein in the KCs increased evidently when stimulated by various concentrations of LPS, and the CD14mRNA expression was correlated in dose-dependent manner with LPS levels. (2) The expression of CD14mRNA and the synthesis of CD14 protein in KCs induced by LPS (10 micro g/ml) increased significantly and peaked at 3 approximately 6 hours. (3) The expression of CD14mRNA and the synthesis of CD14 protein in freshly cultured KCs were obviously up-regulated by the active mediators produced by KCs after being stimulated by LPS. (4) The release of TNFalpha, IL-6 and NO from cultured KCs was evidently down-regulated by the addition of anti-CD14McAb in the presence of serum or by the addition of LPS in the absence of serum, but up-regulated by the concomitant addition of LPS and LBP. CONCLUSION: (1) The CD14mRNA expression and the protein synthesis in cultured KCs were closely related to LPS and the active mediators produced from the KCs.The increased CD14 expression was possibly caused by LPS, and the further increase of the expression might be closely correlated to the cytokines released from the KCs. (2) The KC activation by low concentration of LPS was CD14 dependent.

Experimental study on early liver injury and expressions of TNF-alpha mRNA in burn rats with endotoxemia.

OBJECTIVE: To observe tissue distribution and cell localization of TNF-alpha mRNA and its protein and study their role in the pathogenesis of liver injury in burn rats. METHODS: An animal model of rats subjected to 20% TBSA III degree burns combined with intraperitoneal injection of lipopolysaccharide (LPS) was used for this experiment. The changes of hepatic morphology and functions and serum TNF-alpha content and expression and localization of liver TNF-alpha and TNF-alpha mRNA were determined with light microscope (LM) and electron microscope (EM), quantitative analysis, immunohistochemistry (IHC) and in situ hybridization (ISH). RESULTS: It showed that there were sinusoid reaction, KCs activation and degeneration, necrosis of HCs, and platelets aggregation, fibrins deposition and PMNs attachment in sinusoid. The activity of ALT was obviously elevated and ALB content was slightly decreased. The serum content of TNF-alpha showed peak at 3 hours. TNF-alpha was mainly localized in sinusoid endothelial cells (SECs) and Kupffer cells (KCs), and TNF-alpha mRNA was mainly distributed in KCs, polymorphonuclears neutrophils (PMNs) and macrophages (MPs). CONCLUSIONS: It suggests that TNF-alpha mRNA and its protein expression and localization are coincident with the pathological changes of liver injury. TNF-alpha is one of the key cytokines in the pathogenesis of liver injury in burn rats with endotoxemia.