In order to verify the quantitative trait loci for rice heading time in Koshihikari, QTL mapping of heading time was studied using both Koshihikari/Kasalath BILs population and Kasalath/Koshihikari CSSLs population in Nanjing, Jiangsu province and Lingshui, Hainan province.The results indicated that three QTL of heading time were detected, which were located on chromosomes 3, 6, and 8, respectively.At qHd-3 and qHd-8 loci, the allele from Koshihikari can promote heading time for rice, but at qHd-6-1 loci, the allele from Koshihikari could delay heading time for rice.Epistatic interaction was detected between qHd-3 and the QTL located on chromosome 7. Comparing BILS with CSSLs linkage maps, we found that the marker regions of qHd-3, qHd-8, and qHd-7-1 coincided with Kasalath segment in W008, W023 and W024, W020, respectively.Meanwhile, QTL mapping of heading time were studied using Koshihikari/Guichao 2 RILs population, three additive QTL were detected and epistatic interaction was detected between two unadditive QTL of heading time.
An immunoaffinity column purification-high performance liquid chromatographic method was developed for the determination of vitamin B12 in milk powder.The sample was dissolved with phosphate buffer,and purified through the vitamin B12 immunity affinity column.After eluted with methanol,and concentrated,the target compound was directly measured on a Symmstry C18 column.The optimal conditions were as follows:column temperature:35 ℃,mobile phase:0.03 mol/L potassium dihydrogen phosphate(phosphate adjustable pH 3.0)-acetonitrile(85 ∶ 15),flow rate:1.0 mL/min,detection wavelength:361 nm.Under the optimal conditions,the calibration curve was linear in the range of 20-300 μg/L,with correlation coefficient(r) of 0.999 7 and a detection limit of 0.6 μg/kg.The spiked recoveries were in the range of 90%-96% with RSD of 1.1%-2.4%.Compared with the national standard method,the proposed method has the advantages of easy sample handling,high sensitivity,good reproducibility and short analysis time,and could meet the requirement for the determination of trace amounts of vitamin B12 in milk powder and infant formula.
Background: Skeletal muscle wasting is a common complication of sepsis, leading to severe muscle weakness, atrophy, and a poor prognosis. Studies have shown that programmed cell death is a critical mechanism in sepsis-associated muscular atrophy. Therefore, we aimed to study the role and mechanism of pyroptosis executive protein GSDMD in established cell and mouse models of sepsis.Methods: Sepsis was established by cecal ligation and puncture in Gsdmd knockout and wild-type mice. The atrophy of the gastrocnemius muscle and tibialis anterior muscle was evaluated in a mouse model of sepsis. Skeletal muscle morphology, protein expression, and muscular atrophy-related pathway markers were also analyzed. In vitro, Gsdmd was knocked down in C2C12 cells using siRNA, and the cells were stimulated with lipopolysaccharide (LPS) to assess the effects of Gsdmd downregulation on muscle atrophy and corresponding signaling pathways. Results: Here, we identified an essential role for the "executive" protein of pyroptosis, GSDMD, in skeletal muscle wasting in septic mice. Compared with the control group, the expression of N-GSDMD was significantly increased in the skeletal muscle of septic mice. Compared with septic mice, survival, muscle strength, and body weight were significantly improved in the Gsdmd knockout mice. Gsdmd knockout alleviated gastrocnemius and tibialis anterior muscle atrophy induced by sepsis.Conclusion: This study revealed that Gsdmd knockout ameliorated septic skeletal muscle atrophy by suppressing IL18/AMPK activation and inhibiting the UPS and autophagy.
Malignant glioma is the most common primary tumor of the central nervous system. Chemotherapy and radiotherapy are the most common therapeutic approaches in glioma therapy. Both processes mainly kill cancer cells through generating high Reactive Oxygen Species (ROS) and lead to oxidative DNA damage. However, tumor resistance to ROS is always a challenge for cancer treatment. Human Mut T homolog 1 (MTH1, also known as NUDT1) is regarded as a protector of nucleotides against oxidization. Recent reports have verified that overexpression of MTH1 could remove oxidized dNTP pools. Here, we find that MTH1 is overexpressed both at mRNA and protein levels in GBM. MTH1 silencing inhibits colony formation; tumor spheres formation and xenograft tumor growth, and more importantly, the viability of glioma cells is significantly decreased in H2O2 after MTH1 was knocked down in glioma. PI staining show that H2O2 cause more glioma cell death after MTH1 silencing. So we speculate that overexpression of MTH1 is crucial for glioma survival, suppression of its expression can inhibit cancer cell survival in vitro and in vivo, MTH1 may be a potential target for human glioma therapy in future.
RNA editing generates genetic diversity in mammals by altering amino acid sequences, miRNA targeting site sequences, influencing the stability of targeted RNAs, and causing changes in gene expression. However, the extent to which RNA editing affect gene expression via modifying miRNA binding site remains unexplored. Here, we first profiled the dynamic A-to-I RNA editome across tissues of Duroc and Luchuan pigs. The RNA editing events at the miRNA binding sites were generated. The biological function of the differentially edited gene in skeletal muscle was further characterized in pig muscle-derived satellite cells. RNA editome analysis revealed a total of 171,909 A-to-I RNA editing sites (RESs), and examination of its features showed that these A-to-I editing sites were mainly located in SINE retrotransposons PRE-1/Pre0_SS element. Analysis of differentially edited sites (DESs) revealed a total of 4,552 DESs across tissues between Duroc and Luchuan pigs, and functional category enrichment analysis of differentially edited gene (DEG) sets highlighted a significant association and enrichment of tissue-developmental pathways including TGF-beta, PI3K-Akt, AMPK, and Wnt signaling pathways. Moreover, we found that RNA editing events at the miRNA binding sites in the 3′-UTR of HSPA12B mRNA could prevent the miRNA-mediated mRNA downregulation of HSPA12B in the muscle-derived satellite (MDS) cell, consistent with the results obtained from the Luchuan skeletal muscle. This study represents the most systematic attempt to characterize the significance of RNA editing in regulating gene expression, particularly in skeletal muscle, constituting a new layer of regulation to understand the genetic mechanisms behind phenotype variance in animals. Abbreviations: A-to-I: Adenosine-to-inosine; ADAR: Adenosine deaminase acting on RNA; RES: RNA editing site; DEG: Differentially edited gene; DES: Differentially edited site; FDR: False discovery rate; GO: Gene Ontology; KEGG: Kyoto Encyclopaedia of Genes and Genomes; MDS cell: musclederived satellite cell; RPKM: Reads per kilobase of exon model in a gene per million mapped reads; UTR: Untranslated coding regions
Objective
To investigate the application value in the diagnosis of prostate biopsy tissue frozen section with rapid immunohistochemical detection of p40(ΔNp63),34βE12,p504s.
Methods
The expression of p40, 34βE12,p504s in 118 cases of prostate biopsy tissue frozen section detected by Max Vision rapid immunohistochemical staining technique. Conventional paraffin section immunohistochemistry of frozen remaining tissue served as a control.
Results
118 cases of prostate biopsy histopathological diagnosis was divided into 79 cases of benign prostatic hyperplasia and 39 cases of prostate cancer. Max Vision rapid immunohistochemical staining technique was completed in 30minutes.Antibody expression located accurately. Background stained clearly and no interfering signal.The background was better than conventional immunohistochemistry of frozen remaining tissue. The positive expression rate of p40,34βE12,p504s with rapid immunohistochemistry in prostatic hyperplasia was 97.4%(77/79),93.7%(74/79), 0%(0/79),and in prostatic carcinoma was 0%(0/39),0%(0/39),97.4%(38/39).The positive expression rate of p40,34βE12,p504s with conventional immunohistochemistry in prostatic hyperplasia was 96.2%(76/79),93.7%(74/ 79),2.5%(2/79),and in prostatic carcinoma was 0%(0/39),0%(0/39),92.3%(36/39).The difference of expression between prostatic hyperplasia and prostatic carcinoma with rapid immunohistochemical detection p40 group:χ2= 109.402,P=0.000,34βE12 group:χ2=97.971,P=0.000,p504s group:χ2=113.537,P=0.000;The difference of expression between prostatic hyperplasia and prostatic carcinoma with conventional immunohistochemical detection p40 group:χ2=105.410,P=0.000,34βE12 group:χ2=97.971,P=0.000,p504s group:χ2=96.388,P=0.000;The expression of prostatic hyperplasia with between rapid immunohistochemical detection and conventional immunohistochemical detection 34βE12 group was identical,p40 group:χ2=0.207,P=0.649,p504s group:χ2=2.026,P=0.155; The expression of conventional immunohistochemical detection with between rapid immunohistochemical detection and conventional immunohisto-chemical detection p40 group and 34βE12 group were identical,p504s group:χ2=1.054, P=0.305.The expression of three markers between prostatic hyperplasia and prostatic carcinoma had statistical significance(P 0.05).
Conclusion
Max Vision rapid immunohisto-chemical staining technique has the advantages of rapid, accurate, timely. It could be used to rapid diagnosis of prostate biopsy tissue.The combined detection of p40,34βE12,p504s has very high practical value in the differential diagnosis of benign and malignant lesions of the prostate.
Key words:
Rapid immunohistochemistry; Prostate; p40; 34βE12; p504s; Diagnosis
Glioblastoma multiforme (GBM) is the most common primary malignant tumors originating in the brain parenchyma. At present, GBM patients have a poor prognosis despite the continuous progress in therapeutic technologies including surgery, radiotherapy, photodynamic therapy, and chemotherapy. Recent studies revealed that miR-101 was remarkably down-regulated in kinds of human cancers and was associated with aggressive tumor cell proliferation and stem cell self-renewal. Data also showed that miR-101 was down-regulated in primary glioma samples and cell lines, but the underlying molecular mechanism of the deregulation of miR-101 in glioma remained largely unknown. In this study, we found that miR-101 could inhibit the proliferation and invasion of glioma cells both in vitro and in vivo by directly targeting SOX9 [sex-determining region Y (SRY)-box9 protein]. Silencing of SOX9 exerted similar effects with miR-101 overexpression on glioma cells proliferation and invasion. Quantitative reverse transcription PCR and Western blotting analysis revealed a negative relationship between miR-101 and SOX9 in human glioma U251MG and U87MG cells, and the luciferase assay indicated that miR-101 altered SOX9 expression by directly targeting on 3'UTR. Taken together, our findings suggest that miR-101 regulates glioma proliferation, migration and invasion via directly down-regulating SOX9 both in vitro and in vivo, and miR-101 may be a potential therapeutic target for future glioma treatment.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)