Assessment of Axial Power Peaking Factors in GHARR-1 LEU Core: A Decadal Simulation Analysis

This study aims to thoroughly investigate the axial power peaking factors (PPF) within the low-enriched uranium (LEU) core of the Ghana Research Reactor-1 (GHARR-1). This study uses advanced simulation tools, like the MCNPX code for analysing neutron behavior and the PARET/ANL code for understanding power variations, to get a clearer picture of the reactor’s performance. The analysis covers the initial six years of GHARR-1’s operation and includes projections for its whole 60-year lifespan. We closely observed the patterns of both the highest and average PPFs at 21 axial nodes, with measurements taken every ten years. The findings of this study reveal important patterns in power distribution within the core, which are essential for improving the safety regulations and fuel management techniques of the reactor. We provide a meticulous approach, extensive data, and an analysis of the findings, highlighting the significance of continuous monitoring and analysis for proactive management of nuclear reactors. The findings of this study not only enhance our comprehension of nuclear reactor safety but also carry significant ramifications for sustainable energy progress in Ghana and the wider global context. Nuclear engineering is essential in tackling global concerns, such as the demand for clean and dependable energy sources. Research on optimising nuclear reactors, particularly in terms of safety and efficiency, is crucial for the ongoing advancement and acceptance of nuclear energy.

Insulin-like growth factor binding protein related protein 1 knockdown attenuates hepatic ?brosis via the regulation of MMPs/TIMPs in mice

Background: Previous research suggested that insulin-like growth factor binding protein related protein 1(IGFBPrP1), as a novel mediator, contributes to hepatic fibrogenesis. Matrix metalloproteinases(MMP) and tissue inhibitors of metalloproteinases(TIMP) play an essential role in hepatic fibrogenesis by regulating homeostasis and remodeling of the extracellular matrix(ECM). However, the interaction between IGFBPrP1 and MMP/TIMP is not clear. The present study was to knockdown IGFBPrP1 to investigate the correlation between IGFBPrP1 and MMP/TIMP in hepatic fibrosis. Methods: Hepatic fibrosis was induced by thioacetamide(TAA) in mice. Knockdown of IGFBPrP1 expression by ultrasound-targeted microbubble destruction-mediated CMB-shRNA-IGFBPrP1 delivery, or inhibition of the Hedgehog(Hh) pathway by cyclopamine treatment, was performed in TAA-induced liver fibrosis mice. Hepatic fibrosis was determined by hematoxylin and eosin and Sirius red staining. Hepatic expression of IGFBPrP1, α-smooth muscle actin( α-SMA), transforming growth factor β 1(TGF β1), collagen I, MMPs/TIMPs, Sonic Hedgehog(Shh), and glioblastoma family transcription factors(Gli1) were investigated by immunohistochemical staining and Western blotting analysis. Results: We found that hepatic expression of IGFBPrP1, TGF β1, α-SMA, and collagen I were increased longitudinally in mice with TAA-induced hepatic fibrosis, concomitant with MMP2/TIMP2 and MMP9/TIMP1 imbalance and Hh pathway activation. Knockdown of IGFBPrP1 expression, or inhibition of the Hh pathway, reduced the hepatic expression of IGFBPrP1, TGF β1, α-SMA, and collagen I and re-established MMP2/TIMP2 and MMP9/TIMP1 balance. Conclusions: Our findings suggest that IGFBPrP1 knockdown attenuates liver fibrosis by re-establishing MMP2/TIMP2 and MMP9/TIMP1 balance, concomitant with the inhibition of hepatic stellate cell activation, down-regulation of TGF β1 expression, and degradation of the ECM. Furthermore, the Hh pathway mediates IGFBPrP1 knockdown-induced attenuation of hepatic fibrosis through the regulation of MMPs/TIMPs balance.

Effects of transfected adenovirus-mediated transcription factor X-box binding protein 1 on hippocampal-derived neural stem cell proliferation and apoptosis under hypoxia

BACKGROUND： Neural stem cell （NSC） survival is closely associated with cell apoptosis in ischemic-hypoxic regions following transplantation. Numerous studies have revealed that X-box binding protein 1 （XBP1） is a transcription factor during endoplasmic reticulum unfolded protein response and is essential for cell survival, differentiation, and anti-apoptotic effects. OBJECTIVE： To determine the effects of the XBP1 gene on NSC proliferation and apoptosis under hypoxic conditions following XBP1 gene transfection into rat embryonic hippocampal NSCs using recombinant adenovirus vector. DESIGN, TIME AND SETTING： In vitro experiments were performed at the Laboratory of Cell Biology of Jilin University and Laboratory of Proteomics, Department of Neurology, Jilin University China from September 2008 to November 2009. MATERIALS： Recombinant adenovirus package XBP1 gene and Ad-XBPl-enhanced green fluorescent protein plasmid （Guangzhou Easywin BioMed Technology, China）, rabbit anti-XBP1 and its target gene estrogen receptor degradation-enhancing a-mannosidase-like protein （EDEM） glucose-regulated protein 78 （GRP78）, anti-apoptotic molecule Bcl-2 and proapoptotic molecule Bax polyclonal antibody （Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA）, and COCI2 （Sigma, St. Louis, MO, USA） were used in the present study. METHODS： Hippocampi from embryonic, Sprague Dawley rats on gestational day 16 were harvested for NSC isolation and cloning, followed by immunofluorescence for Nestin and sub-culturing. The recombinant adenovirus Ad-XBPl-enhanced green fluorescent protein plasmid was transfected into rat embryonic hippocampal NSCs, and then CoCl2 was applied to induce hypoxia. MAIN OUTCOME MEASURES： Cell quantification and 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide colorimetric assay were utilized to detect proliferation in XBPl-transfected NSCs for 7 consecutive days. Western blot assay was utilized to quantify XBP1 GRP78, EDEM, Bcl-2, and Bax expression. Flow cytometry was used to measure apoptosis. RESULTS： NSC proliferation was significantly enhanced following XBP1 gene transfection （P 〈 0.05）. Under hypoxic conditions, GRP78, EDEM, and Bcl-2 levels increased, but Bax levels decreased. In addition, NSC apoptosis decreased following transfection （P 〈 0.05）. CONCLUSION： The XBP1 gene was successfully transfected into rat embryonic hippocampal NSCs using a recombinant adenovirus vector. NSC proliferation following transfection, as well as anti-apoptotic effects under hypoxia, was significantly increased.

Interaction between insulin-like growth factor binding protein-related protein 1 and transforming growth factor beta 1 in primary hepatic stellate cells

BACKGROUND: We previously showed that insulin-like growth factor binding protein-related protein 1 (IGFBPrP1) is a novel mediator in liver fibrosis. Transforming growth factor beta 1 (TGF beta 1) is known as the strongest effector of liver fibrosis. Therefore, we aimed to investigate the detailed interaction between IGFBPrP1 and TGF beta 1 in primary hepatic stellate cells (HSCs). METHODS: We overexpressed TGF beta 1 or IGFBPrP1 and inhibited TGF beta 1 expression in primary HSCs for 6, 12, 24, 48, 72, and 96 hours to investigate their interaction and observe the accompanying expressions of a-smooth muscle actin (alpha-SMA), collagen I, fibronectin, and phosphorylated-mothers against decapentaplegic homolog 2/3 (p-Smad2/3). RESULTS: We found that the adenovirus vector encoding the TGF beta 1 gene (AdTGF beta 1) induced IGFBPrP1 expression while that of alpha-SMA, collagen I, fibronectin, and TGF beta 1 increased gradually. Concomitantly, AdIGFBPrP1 upregulated TGF beta 1, alpha-SMA, collagen I, fibronectin, and p-Smad2/3 in a time-dependent manner while IGFBPrP1 expression was decreased at 96 hours. Inhibition of TGF beta 1 expression reduced the IGFBPrP1-stimulated expression of alpha-SMA, collagen I, fibronectin, and p-Smad2/3. CONCLUSIONS: These findings for the first time suggest the existence of a possible mutually regulation between IGFBPrP1 and TGF beta 1, which likely accelerates liver fibrosis progression. Furthermore, IGFBPrP1 likely participates in liver fibrosis in a TGF beta 1-depedent manner, and may act as an upstream regulatory factor of TGF beta 1 in the Smad pathway.

Study on the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell lines

Objective： Detecting the expression and mutation of human telomeric repeat binding factor （hTRF1） in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes to clarify ifhTRF1 mutation is one of the factors of the activation of telomerase. Methods： hTRFlcDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of h TRFlmRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-I, HEL and Dami; lymphoblastic leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification （TRAP）-ELISA protocol. PCR and sequencing were used to detect mutation of each exon ofhTRF1 in 10 cell line cells. Results： hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes ofhTRF1 located on chromosome 13, 18, 21 and X respectively, was named as ψhTRFI-13, ψhTRFI-18, ψhTRF1-21 and ψhTRFI-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells （0.0338, 0.0108-0.0749） than in normal mononuclear cells （0.0493, 0.0369-0.128） （P=0.004）. But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part ofintron 1, 2 and 8 ofhTRF1. Their infection on gene function is unknown and needs further studies. Conclusion： hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease.

Insulin-like growth factor 2 mRNA-binding protein 1 promotes cell proliferation via activation of AKT and is directly targeted by microRNA-494 in pancreatic cancer

BACKGROUND Studies have shown that insulin-like growth factor 2 mRNA-binding protein 1(IGF2BP1)plays critical roles in the genesis and development of human cancers.AIM To investigate the clinical significance and role of IGF2BP1 in pancreatic cancer.METHODS Expression levels of IGF2BP1 and microRNA-494(miR-494)were mined based on Gene Expression Omnibus datasets and validated in both clinical samples and cell lines by quantitative real-time polymerase chain reaction and Western blot.The relationship between IGF2BP1 expression and clinicopathological factors of pancreatic cancer patients was analyzed.The effect and mechanism of IGF2BP1 on pancreatic cancer cell proliferation were investigated in vitro and in vivo.Analyses were performed to explore underlying mechanisms of IGF2BP1 upregulation in pancreatic cancer and assays were carried out to verify the posttranscriptional regulation of IGF2BP1 by miR-494.RESULTS We found that IGF2BP1 was upregulated and associated with a poor prognosis in pancreatic cancer patients.We showed that downregulation of IGF2BP1 inhibited pancreatic cancer cell growth in vitro and in vivo via the AKT signaling pathway.Mechanistically,we showed that the frequent upregulation of IGF2BP1 was attributed to the downregulation of miR-494 expression in pancreatic cancer.Furthermore,we discovered that reexpression of miR-494 could partially abrogate the oncogenic role of IGF2BP1.CONCLUSION Our results revealed that upregulated IGF2BP1 promotes the proliferation of pancreatic cancer cells via the AKT signaling pathway and confirmed that the activation of IGF2BP1 is partly due to the silencing of miR-494.

Significance of highly phosphorylated insulin-like growth factor binding protein-1 and cervical length for prediction of preterm delivery in twin pregnancies

BACKGROUND A twin pregnancy can carry greater risks than singleton pregnancies.About 60 in 100 twin pregnancies result in spontaneous birth before 37 wk,which is associated with several complications in the premature babies.Clinical detection of biomarkers may help to predict the possibility of premature birth so that corresponding interventions can be given to the pregnant women in a timely manner,in order to reduce the risk of preterm birth and improve the outcomes of the newborn infants.AIM To explore the clinical value of transvaginal ultrasound measurement of cervical length combined with insulin-like growth factor binding protein-1(IGFBP-1)hyperphosphorylation in cervical secretions as predictors of preterm delivery in twin pregnancies.METHODS A total of 254 pregnant women with twin pregnancies,who were admitted to Hainan General Hospital and underwent maternity examination,were selected as the study subjects from January 2015 to December 2018.All participants received transvaginal ultrasound measurement of cervical length and phosphorylated IGFBP-1(phIGFBP-1)test between 24 and 34 wk gestation.The pregnancy outcomes were analyzed.RESULTS Of the women with a positive phIGFBP-1 test result,preterm birth rate was higher in those with a cervical length≤25 mm than those with a cervical length>25 mm(all P<0.05).Similarly,in women with a negative phIGFBP-1 test result,preterm birth rate was higher in those with a cervical length≤25 mm than those with a cervical length>25 mm(all P<0.05).The sensitivity,specificity,and positive and negative predictive values of the phIGFBP-1 test combined with the cervical length test were 95.71%,91.21%,95.12%and 92.22%,respectively,for the prediction of preterm birth.CONCLUSION Cervical length combined with phIGFBP-1 tests is of value for the prediction of outcomes of preterm delivery in twin pregnancies.

Insulin-like growth factor-binding protein-3 inhibits IGF-1-induced proliferation of human hepatocellular carcinoma cells

OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.

Tribulus terrestris extracts alleviate muscle damage and promote anaerobic performance of trained male boxers and its mechanisms： Roles of androgen, IGF-1, and IGF binding protein-3

Purpose: To investigate the effects of Tribulus terrestris(TT) extracts on muscle mass, muscle damage, and anaerobic performances of trained male boxers and its mechanisms: roles of plasma androgen, insulin growth factor 1(IGF-1), and IGF-1 binding protein-3(IGFBP-3).Methods: Fifteen male boxers were divided into exercise group(E, n = 7) and exercise plus TT group(E + TT, n = 8). The 2 groups both undertook3-week high-intensity and 3-week high-volume trainings separated by a 4-week rest. TT extracts(1250 mg/day) were orally administered by boxers in E + TT group. TT extract compositions were detected by UHPLC–Q-TOF/MS. Before and at the end of the 2 trainings, muscle mass, anaerobic performance, and blood indicators were explored.Results: Compared with E group, decreases of plasma CK(1591.5 ± 909.6 U/L vs. 2719.9 ± 832.5 U/L) and IGFBP-3(3075.5 ± 1072.5 ng/m L vs. 3950.8 ± 479.3 ng/m L) as well as increases of mean power(MP, 459.4 ± 122.3 W vs. 434.6 ± 69.5 W) and MP/body weight(MP/BW, 7.5 ± 0.9 W/kg vs. 7.1 ± 1.1 W/kg) were detected in E + TT group after a high-intensity training. For high-volume training, reduction of IGFBP-3(2946.4 ± 974.1 ng/m L vs. 3632.7 ± 470.1 ng/m L) and increases of MP(508.7 ± 103.2 W vs. 477.8 ± 49.9 W) and MP/BW(8.2 ± 0.3 W/kg vs.7.5 ± 0.9 W/kg) were detected in E + TT group, compared with E group. Muscle mass, blood levels of testosterone, dihydrotestosterone(DHT),and IGF-1 were not signifiantly changed between the 2 groups.Conclusion: Taking 1250 mg capsules containing TT extracts did not change muscle mass and plasma levels of testosterone, DHT, and IGF-1 but significantly alleviated muscle damage and promoted anaerobic performance of trained male boxers, which may be related to the decrease of plasma IGFBP-3 rather than androgen in plasma.

抑制NRF1/ABCC1提高人肺腺癌细胞系对顺铂化学治疗的敏感性

目的探讨核呼吸因子1(NRF-1)激活ATP结合盒转运蛋白C1(ABCC1)转录对肺腺癌细胞顺铂抵抗的影响及其潜在机制。方法通过癌基因组数据集分析肺腺癌肿瘤组织中ABCC1的表达水平。细胞分组:sh-NC、sh-ABCC1、sh-NC+oe-NC、sh-NRF1+oe-NC和sh-NRF1+oe-ABCC1。RT-qPCR检测ABCC1在肺腺癌细胞中的表达量。构建顺铂耐药肺腺癌细胞株,检测ABCC1的表达水平。(0、0.001,0.002,0.004、0.008、0.016和0.032 mg/mL)顺铂处理的耐药肺腺癌细胞的IC50值。Western blot检测上皮间充质转化标志物(E-cadherin、N-cadherin)蛋白的表达。双荧光素酶报告基因和ChIP实验验证NRF1与ABCC1的结合关系。结果ABCC1在肺腺癌肿瘤组织和细胞中高表达。与顺铂敏感的肺腺癌细胞相比,ABCC1在顺铂耐药肺腺癌细胞中高表达,敲低ABCC1可显著抑制细胞增殖、迁移和侵袭,并提高E-cadherin的表达(P<0.05),降低N-cadherin的表达(P<0.05)。敲低ABCC1可显著提高肺腺癌细胞对顺铂的敏感性(P<0.05)。此外,双荧光素酶报告基因和ChIP实验证实NRF1与ABCC1启动子的去结合关系,且NRF1可激活ABCC1的转录。敲低NRF1可减弱过表达ABCC1对肺腺癌细胞增殖、迁移、侵袭及顺铂抵抗的抑制作用(P<0.05)。结论NRF1/ABCC1轴在肺腺癌顺铂抵抗中具有促进作用。抑制NRF1/ABCC1轴可能是提高肺腺癌顺铂敏感性的潜在靶点。

miR-192-5p靶向CKIP-1促进骨质疏松患者骨髓间充质干细胞成骨分化

背景:酪蛋白激酶2结合蛋白1(casein kinase 2-interaction protein-1,CKIP-1)是一种重要的骨形成负调控基因,其敲除鼠骨质显著增强、骨形成和骨密度也显著提高。而miRNA作为较早发现的小分子调控物,对大多数编码基因具有调控作用,在成骨分化中发挥重要作用。目的:探讨miRNA/CKIP-1对骨质疏松患者骨髓间充质干细胞成骨分化的影响及其分子机制。方法:采用miRNA-Seq技术检测2022年3-6月在开封市中心医院骨外科就诊32例骨质疏松患者及同期体检中心健康人群骨髓间充质干细胞中miRNA的变化情况;利用Targetscan网站预测靶向调控CKIP-1的miRNA,利用荧光素酶报告基因实验检测miRNA与CKIP-1启动子区DNA的结合;在骨髓间充质干细胞中转染miR-192-5p类似物(miR-192-5p mimics)/阴性对照(NC mimics)或miR-192-5p抑制剂(miR-192-5p inhibitor)/阴性对照(NC inhibitor),成骨诱导后第7,14天,通过实时荧光定量PCR技术及茜素红染色检测成骨标志基因Runt相关转录因子2(Runx2)、骨钙素、抗骨桥蛋白、骨唾液蛋白及CKIP-1的表达水平和骨髓间充质干细胞向成骨细胞分化的情况;采用蛋白质免疫印迹实验及茜素红染色检测miR-192-5p/CKIP-1/轴对细胞成骨分化的的调控作用。结果与结论:与健康组相比,骨质疏松组有16个miRNA表达明显升高,53个miRNA表达明显降低(P<0.05);利用Targetscan网站预测,并通过荧光素酶报告基因实验验证,发现miR-192-5p与CKIP-1有互补的核苷酸序列(P<0.05);过表达miR-192-5p,Runx2、骨钙素、骨桥素和骨唾液蛋白的表达水平显著升高(P<0.05),抑制miR-192-5p,Runx2、骨钙素、骨桥素和骨唾液蛋白的表达水平显著降低(P<0.05),而沉默CKIP-1的表达后,Runx2、骨钙素及骨桥素的蛋白水平增加(P<0.05),逆转了敲低miR-192-5p对细胞成骨分化的抑制作用。上述结果证实,miR-192-5p在骨质疏松症中表达降低;miR-192-5p通过靶向抑制CKIP-1的表达,促进骨髓间充质干细胞成骨分化。

Extracellular and cytoplasmic regions of LRIG1 play a negative role in EGFR activity: Findings of a radioligand-binding assay

Objective Leucine-rich repeats and immunoglobulin-like domains 1(LRIG1) is a newly identified human gene that inhibits the epidermal growth factor receptor(EGFR), which on combining with a ligand, can drive tumor growth. This study investigated the interaction between human LRIG1 and EGFR and attempted to delineate the functions of as well as the mechanisms used by the extracellular(ECD) and cytoplasmic(CPD) domains of the human LRIG1 protein to downregulate human EGFR signaling activity.Methods Two constructed chimeric eukaryotic expression vectors, pIRES2-EGFP-3XFLAG-LRIG1-ET and p3FLAG-LRIG1-TC, encoding the extracellular and transmembrane regions(LRIG1-ET) and the transmembrane and cytoplasmic regions(LRIG1-TC), respectively, and the plasmid p3XFLAG-CMV-9-LRIG1 encoding full-length LRIG1(LRIG1-FL) were transfected into the human glioma cell line U251 or primary astrocytoma cells by using liposomes. The number and affinity of cell surface EGFR on transfected cells was determined by ^(125)I-EGF binding assay. Results The dissociation constant(KD) values for EGFR were higher, and the maximum increase was observed in the cells transfected into LRIG1-ET(1.36 folds). The number of maximal binding sites(Bmax) of the receptors was decreased in all transfected cells; the maximum decrease was noted in the cells transfected into LRIG1-FL(40.05%).Conclusion Both the ECD and CPD of LRIG1 are important to negate EGFR signaling. The ECD may interfere with the binding between EGFR and its ligand and facilitate the functions of CPD. The CPD may, when brought in proximity to EGFR, enhance receptor degradation. These two mechanisms can contribute to the downregulation of EGFR-mediated signaling by LRIG1.

A distribution pattern assisted method of transcription factor binding site discovery for both yeast and filamentous fungi

Transcription factors (TFs) are the core sentinels of gene regulation functioning by binding to highly specific DNA sequences to activate or repress the recruitment of RNA polymerase. The ability to identify transcription factor binding sites (TFBSs) is necessary to understand gene regulation and infer regulatory networks. Despite the fact that bioinformatics tools have been developed for years to improve computational identification of TFBSs, the accurate prediction still remains changeling as DNA motifs recognized by TFs are typically short and often lack obvious patterns. In this study we introduced a new attribute-motif distribution pattern (MDP) to assist in TFBS prediction. MDP was developed using a TF distribution pattern curve generated by analyzing 25 yeast TFs and 37 of their experimentally validated binding motifs, followed by calculating a scoring value to quantify the reliability of each motif prediction. Finally, MDP was tested using another set of 7 TFs with known binding sites to in silico validate the approach. The method was further tested in a non-yeast system using the filamentous fungus Magnaporthe oryzae transcription factor MoCRZ1. We demonstrate superior prediction reranking results using MDP over the commonly used program MEME and the other four predictors. The data showed significant improvements in the ranking of validated TFBS and provides a more sensitive statistics based approach for motif discovery.

终末期肾脏病患者桡动脉中膜钙化与核心结合因子α_1及Ⅰ型胶原的关系研究

目的探讨终末期肾脏病（ESRD）患者桡动脉中膜钙化与核心结合因子α1（Cbfα1）及Ⅰ型胶原的关系。方法选择2009年5月—2012年12月在河北医科大学第四医院肾内科住院初次行前臂动-静脉内瘘成形术的ESRD患者62例。采用von kossa染色法观察桡动脉钙化情况,并根据组织钙化积分将患者分为无钙化组（〈1.0分）和钙化组（1.0~4.0分）;采用免疫组织化学法观察Cbfα1及Ⅰ型胶原的表达情况;检测相关临床指标〔空腹血清钙（Ca）、血清磷（P）、三酰甘油（TG）、总胆固醇（TC）、低密度脂蛋白（LDL）、高密度脂蛋白（HDL）、血红蛋白（Hb）、碱性磷酸酶（ALP）、铁蛋白、甲状旁腺素（iPTH）和C反应蛋白（CRP）,血压,体质指数（BMI）〕。结果62例ESRD患者桡动脉标本中25例（40.3%）发生血管钙化,其中轻中度钙化22例（35.5%）,重度钙化3例（4.8%）,均发生在血管中膜。根据组织钙化积分,无钙化组37例,钙化组25例。钙化组桡动脉中膜均有Cbfα1及Ⅰ型胶原的阳性表达;无钙化组桡动脉中膜31例（83.8%）Cbfα1阳性表达,19例（51.4%）Ⅰ型胶原阳性表达。钙化组Cbfα1及Ⅰ型胶原的阳性表达积分均高于无钙化组（P〈0.05）。钙化组与无钙化组的年龄、BMI、血压、Ca、iPTH、TG、TC、LDL、HDL、Hb、CRP、铁蛋白、ALP比较,差异无统计学意义（P〉0.05）;钙化组血清P水平高于无钙化组（P〈0.05）。钙化组Cbfα1阳性表达积分与血清P呈正相关（r=0.679,P〈0.05）,与年龄、BMI、血压、血清Ca、TG、TC、LDL、HDL、iPTH、Hb、CRP、铁蛋白、ALP无相关性（P〉0.05）;Ⅰ型胶原阳性表达积分与血清P呈正相关（r=0.393,P〈0.05）,与年龄、BMI、血压、血清Ca、TG、TC、LDL、HDL、iPTH、Hb、CRP、铁蛋白、ALP无相关性（P〉0.05）;Cbfα1阳性表达积分与Ⅰ型胶原阳性表达积分呈正相关（r=0.307,P〈0.05）。结论 ESRD患者血管中膜钙化发生率较高,其发生机制可能与高磷诱导血管平滑肌细胞表型转化,促进I型胶原表达增加有关,为临床进一步研究提供了参考依据。

补肾中药对增龄大鼠成骨细胞Cbfα_1表达的影响

目的观察核心结合因子α1(Cbfα1)在增龄成骨细胞中的表达,探讨补肾中药促进骨形成的新机制。方法来源于不同年龄的大鼠成骨细胞被分离,采用组织块翻转方法培养。通过倒置显微镜观察细胞形态,矿化结节染色对成骨细胞加以鉴定。采用RT-PCR和免疫组化方法检测成骨细胞Cbfα1的表达。结果RT-PCR和免疫组化结果表明,在增龄成骨细胞中,固本壮骨胶囊、金匮肾气丸和西药萌格旺可上调不同龄大鼠成骨细胞Cbfα1的表达,知柏地黄丸则下调Cbfα1的表达,补肾益精方对于不同龄大鼠成骨细胞Cbfα1表达的影响存在着区别。结论补肾中药中的补阳中药具有促进骨形成的作用。

AdEasy1/Cbfα_1诱导间充质干细胞向成骨细胞分化的实验研究

目的:观察核心结合因子α1(Cbfα1)对兔骨髓间充质干细胞(MSCs)向成骨细胞分化的诱导作用。方法:体外分离培养兔骨髓MSCs,用AdEasy1/Cbfα1转染MSCs,在转染后3 d、1、2、3和4周时,组织化学和免疫组化等方法检测成骨标志碱性磷酸酶和骨钙素的表达。结果:AdEasy1/Cbfα1转染后的兔骨髓MSCs表现出与成骨细胞相似的形态,并且表达碱性磷酸酶和骨钙素。结论:Cbfα1可诱导兔骨髓MSCs向成骨细胞分化。

核心结合因子α_1基因修饰MSCs对骨质疏松大鼠骨形成作用研究

目的:以去势大鼠建立绝经后骨质疏松模型,探讨以核心结合因子α1(Cbfα1)基因修饰骨髓间充质干细胞(MSCs)作为种子细胞,对绝经后骨质疏松症骨形成的刺激作用和基因治疗机制。方法:54只SD雌性大鼠随机分为三组:去势后给药实验组(Cbfα1基因修饰MSCs植入组:(G1),去势组(G2)和对照组(G3),并将术后第4、10和16定为实验周。观测骨形成指标:骨钙素(BGP)、骨特异性碱性磷酸酶(骨AKP)和总碱性磷酸酶(总AKP)和骨吸收指标:尿吡啶醚(PYD)和脱氧吡啶醚(DPD)以及骨生物力学检测(BMT)。同步用IBAS计算机全自动图像分析系统对不脱钙骨组织动态观测骨形态计量学参数。结果:G1在去势后4～16周BGP和骨AKP明显高于G2。而骨吸收指标:尿PYD和DPD两组间无显著差异。骨形成表面(FS)和骨小梁体积比(TBV),骨小梁平均厚度(MTT)较G2明显升高,尤其是TBV和FS,这种差异在第10周最为显著。随着给药时间的延长(第16周)骨矿化沉积速率(MAR)逐渐增加。骨吸收表面(RS)与G2比较在所有实验周均无差异。BMT值4～16周明显高于G2。结论:Cbfα1基因修饰MSCs可刺激骨质疏松大鼠成骨细胞活跃增生,促进骨形成与骨转换速率代谢平衡。并可使骨质疏松大鼠骨组织微观结构改善。

抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用

目的探讨敲低长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)抑制核苷酸结合寡聚结构域样受体蛋白1(NLRP1)炎症小体在缓解阿尔茨海默病进展中的作用。方法选取9~10周龄遗传背景为C57/BL6的野生型小鼠(WT组,10只)或淀粉样前体蛋白(APP)/早老素1(PS1)转基因小鼠(30只)。APP/PS1转基因小鼠随机分为模型(model)组,模型+敲低lncRNA TUG1组[model+lncRNA TUG1短发夹RNA(shRNA)组]和model+shRNA非靶标(NT)组,每组10只。分别采集12周龄第1天(3月龄)和32周龄第1天(8月龄)小鼠外周血和脑皮质组织,并分离皮质中的原代小胶质细胞和原代星形胶质细胞,每个时间点每组5只小鼠。Real-time PCR分别测定3月龄和8月龄上述4个分组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和巨噬细胞移动抑制因子(MIF)mRNA的水平,以及原代星形胶质细胞中补体蛋白C1r和C1s mRNA的水平。ELISA法测定其外周血浆中MIF含量。对3月龄和8月龄小鼠脑皮质原代小胶质细胞和原代星形胶质细胞共培养。CCK-8法测定上述2种细胞的增殖能力。Western blotting分别测定3月龄和8月龄上述4个分组小鼠脑皮质组织中MIF、白细胞介素1β前体(pro-IL-1β)、凋亡相关斑点样蛋白(ASC)、Caspase-1(p20)、Caspase-1(full)、NLRP1及NLRP3蛋白的表达水平。采用免疫荧光染色法测定8月龄各分组小鼠脑皮质组织中β淀粉样蛋白(Aβ)表达。结果3月龄和8月龄时,与WT组小鼠相比,model组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF相对表达水平显著上调,原代小胶质细胞和原代星形胶质细胞增殖能力增强(P<0.05)。与model组相比,model+lncRNA TUG1 shRNA组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF的相对表达水平显著降低,原代小胶质细胞和原代星形胶质细胞增殖能力降低(P<0.05)。与WT组相比,model组小鼠外周血浆中MIF含量显著升高;小鼠脑皮质组织中pro-IL-1β、ASC、Caspase-1(p20)、Caspase-1(full)、NLRP1以及NLRP3的蛋白表达水平显著升高;Aβ免疫荧光强度明显增强(P<0.05)。与model组相比,model+lncRNA TUG1 shRNA组小鼠外周血浆中MIF含量显著降低;小鼠脑皮质组织中pro-IL-1β、ASC、Caspase-1(p20)、Caspase-1(full)和NLRP1的蛋白表达水平显著降低,Aβ免疫荧光强度明显降低(P<0.05),而NLRP3蛋白质的表达水平无明显变化(P>0.05)。与model组相比,model+shRNA NT组小鼠上述所有检测指标差异均无显著性(P>0.05)。结论APP/PS1转基因小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF因子表达上调与脑皮质内NLRP1炎症小体激活成正相关,敲低lncRNA TUG1可缓解阿尔茨海默病的进展。

血清LTBP2、TM4SF1在大肠癌患者预后评估中的研究

目的探讨血清转化生长因子结合蛋白2(LTBP2)、四次跨膜蛋白1(TM4SF1)在大肠癌患者预后评估中的意义。方法将南阳市中心医院2016年7月至2019年7月手术治疗的大肠癌患者108例作为试验组,同期108例健康体检者作为对照组。采用酶联免疫吸附试验(ELISA)检测两组血清LTBP2、TM4SF1水平;比较不同血清LTBP2和TM4SF1表达水平的大肠癌患者临床资料的差异;采用免疫组化染色法分析大肠癌组织中LTBP2、TM4SF1的表达;Pearson相关分析明确血清LTBP2和TM4SF1表达的相关性;受试者工作特征(ROC)曲线分析血清LTBP2和TM4SF1联合评估大肠癌患者预后的价值。结果试验组血清LTBP2和TM4SF1水平均高于对照组(P<0.05),二者表达水平呈正相关(r=0.305,P<0.05),大肠癌组织LTBP2和TM4SF1表达阳性率高于癌旁组织(P<0.05)。肿瘤直径>5cm、肿瘤低分化、有淋巴结转移、TNM分期Ⅲ~Ⅳ期、有脉管瘤栓的大肠癌患者血清LTBP2和TM4SF1表达水平高于患者肿瘤直径≤5 cm、肿瘤中高分化、无淋巴结转移、TNM分期Ⅰ~Ⅱ期、无脉管瘤栓的大肠癌患者(P<0.05)。血清LTBP2、TM4SF1联合预测大肠癌患者预后不良的AUC为0.886。结论大肠癌患者血清LTBP2和TM4SF1水平升高,二者联合对大肠癌患者预后不良具有较好的预测价值。

脑脊液IGF-1、β_(2)-MG、HBP与儿童化脓性脑膜炎发生发展的相关性

目的探究脑脊液中胰岛素样生长因子-1(IGF-1)、β_(2)微球蛋白(β_(2)-MG)、肝素结合蛋白(HBP)水平与儿童化脓性脑膜炎疾病发生发展的相关性。方法选取2021年2月至2023年2月郑州大学第一附属医院儿科收治的86例化脓性脑膜炎患儿为研究组,另选取86例非中枢神经系统感染患儿为对照组,比较两组脑脊液IGF-1、β_(2)-MG、HBP水平,探究其与研究组患儿病情严重程度及预后的相关性。结果研究组脑脊液中IGF-1、β_(2)-MG、HBP水平均高于对照组(P<0.05),且对化脓性脑膜炎具有较高的诊断价值,其曲线下面积分别为0.925、0.930、0.850;重度组脑脊液IGF-1、β_(2)-MG、HBP水平高于轻度组(P<0.05);预后不良组脑脊液IGF-1、β_(2)-MG、HBP水平高于预后良好组(P<0.05),且对化脓性脑膜炎预后不良具有较高的预测价值,其曲线下面积分别为0.879、0.854、0.822。结论监测化脓性脑膜炎患儿脑脊液中IGF-1、β_(2)-MG、HBP的水平对于早期诊断、判断病情严重程度以及预测疾病预后具有重要价值。