Advances on BTB protein ubiquitination mediated plant development and stress response / 生物工程学报

The BTB (broad-complex, tramtrack, and bric-à-brac) domain is a highly conserved protein interaction motif in eukaryotes. They are widely involved in transcriptional regulation, protein degradation and other processes. Recently, an increasing number of studies have shown that these genes play important roles in plant growth and development, biotic and abiotic stress processes. Here, we summarize the advances of these proteins ubiquitination-mediated development and abiotic stress responses in plants based on the protein structure, which may facilitate the study of this type of gene in plants.

Interactive web-based tool for evaluating the spread of bovine tuberculosis and brucellosis in Colombia / Herramienta interactiva basada en la web para evaluar la propagación de la tuberculosis bovina y la brucelosis en Colombia / Ferramenta interativa baseada na web para avaliar a disseminação da tuberculose e brucelose bovina na Colômbia

Abstract Background: Bovine tuberculosis (BTB) and brucellosis are associated with devastating losses in the livestock sector in Colombia and even in developed countries. Real-time disease surveillance is a key strategy to control and eradicate infectious disease outbreaks. Objective: To design an epidemiological tool for monitoring BTB and brucellosis in Colombia. Methods: An interactive platform for disease mapping of BTB and brucellosis during an observation period between years 2004 and 2019 was designed. Results: Our analysis showed that the provinces of Cundinamarca and Valle del Cauca are regions affected by BTB and brucellosis epidemics, respectively (p<0.001). Furthermore, increased case detection of BTB was reported in 2012 and brucellosis in 2019 (p<0.001). Conclusions: This epidemiological platform allows tracking BTB and tuberculosis hotspots, identifying trends over time, and provides useful information to animal health authorities for designing new strategies in control programs.

Resumen Antecedentes: La tuberculosis bovina (TBB) y la brucelosis están asociadas con problemas persistentes en la ganadería Colombiana e incluso en los países desarrollados. La vigilancia de enfermedades en tiempo real es una estrategia clave para controlar y erradicar brotes de enfermedades infecciosas. Objetivo: Diseñar una herramienta epidemiológica para monitorear TBB y brucelosis en Colombia. Métodos: Se diseñó un panel de control interactivo para el mapeo de ambas enfermedades durante el periodo de observación entre los años 2004 y 2019. Resultados: El análisis de la herramienta mostró que las Provincias de Cundinamarca y Valle del Cauca han sido áreas epidémicas para TBB y brucelosis, respectivamente (p<0,001). Además, se encontró un aumento de la detección de casos de TBB en 2012 y de brucelosis durante 2019 (p<0,001). Conclusiones: Este panel epidemiológico permite el seguimiento de puntos críticos de TBB y tuberculosis, identificando sus tendencias a lo largo del tiempo, y proporciona información útil para las autoridades de sanidad animal que diseñan nuevas estrategias para los programas de control.

Resumo Antecedentes: A tuberculose bovina (TBB) e a brucelose estão associadas a problemas persistentes no campo da pecuária na Colômbia e até em países desenvolvidos. Portanto, a vigilância de doenças em tempo real é uma estratégia essencial para controlar e erradicar surtos de doenças infecciosas. Objetivo: Projetar uma ferramenta epidemiológica para monitorar a TB e a brucelose na Colômbia. Métodos: Um painel de controle interativo foi projetado para o mapeamento de ambas as doenças entre 2004 e 2019 como período de observação. Resultados: A análise da ferramenta mostrou que as Províncias de Cundinamarca e Valle del Cauca foram áreas epidêmicas para TBB e brucelose, respectivamente (p<0,001). Além disso, foi encontrado um aumento na detecção de casos em 2012 para TBB e brucelose durante 2019 (p<0,001). Conclusões: Esse painel epidemiológico poderia permitir o monitoramento de pontos críticos dessas doenças, identificando tendências ao longo do tempo, fornecendo informações úteis para as autoridades de saúde animal que elaboram novas estratégias para programas de controle.

The Role of SPOP During Development / 中国生物化学与分子生物学报

Aberrant expression or mutation of many genes that are essential for embryonic development, are closely associated with human diseases, one of which is SPOP (speckle type BTB/POZ protein). SPOP is an E3 ubiquitin ligase adaptor protein and mainly composed of MATH, BTB and BACK domains, which plays distinct roles to fulfill the proper function of SPOP. SPOP usually targets its substrates for degradation via the ubiquitin-proteasome pathway. More than thirty substrates of SPOP have been identified by far, most of which are associated with tumorigenesis of prostate, endometrial and kidney cancers. SPOP also plays an important role during development. Genomic loss or mutation of SPOP locus leads to postnatal lethality in mice, while de novo variants in SPOP cause neurodevelopmental disorders in children. Similarly, SPOP regulates a variety of developmental processes via targeting its substrates for degradation, including Gli2/3, PDX1, NANOG and SENP7 which are involved in neural, skeletal and pancreatic development as well as senescence. In addition, recent studies have revealed that SPOP co-localizes with its substrates into membraneless organelles such as nuclear speckles, and promotes ubiquitination and degradation of its substrates. Oligomerization of SPOP and liquid-liquid phase separation (LLPS) triggered by multivalent interactions between SPOP and substrates play a pivotal role in this process. BTB or BACK mutants, which are defective in SPOP oligomerization, are also defective in driving LLPS of SPOP and recruiting SPOP into membraneless organelles. In this review, we summarized and discussed the recent progress on the essential role of SPOP during development.

Effects of up-regulation of zinc finger and BTB domain containing 20 expression on learning and memory of APPswe/PSE9 transgenic Alzheimer disease mice / 解剖学报

Objective To investigate the effects of up-regulation of zinc finger and BTB domain containing 20 (A20) expression on learning and memory and apoptosis of hippocampal neurons in APP

MALAT1 and BACH1 are prognostic biomarkers for triple-negative breast cancer

Aims: The purpose of this study was to investigate the potential correlation between metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and the transcription factor BTB and CNC homology 1 (BACH1) and their clinicopathological significance in triple-negative breast cancer (TNBC). Subjects and Methods: MALAT1 and BACH1 were detected by immunohistochemistry using TNBC tissue microarrays of 240 patients. The association between MALAT1 and BACH1 expression levels was statistically analyzed. Moreover, the prognostic roles as well as clinical and pathological significance of MALAT1 and BACH1 expression in TNBC were determined. Statistical Analysis Used: Two-tailed Pearson correlation was used to examine the correlation of BACH1 and MALA1 expression. Comparisons of clinicopathological variables between different BACH1 and MALA1 expression groups were performed using χ2 tests. Overall survival (OS) and disease-free survival (DFS) curves were plotted with the Kaplan-Meier method and the differences in OS and DFS between three groups were compared by the log-rank test. Multiple comparisons were performed using χ2 tests for subsequent individual group comparisons. Results: MALAT1 and BACH1 expression was significantly correlated with tumor-node-metastasis stage, distant metastasis, pathological stage, and survival outcomes of patients. Patients with high MALAT1 and BACH1 expression exhibited shorter overall survival and disease-free survival. Conclusions: These findings provide further insight into the expression pattern of MALAT1 and BACH1 in TNBC and suggest them as prognostic biomarkers for TNBC

Expression of transcription factor Bach2 in CD4+CD25+CD45RA- T cells in patients with systemic lupus erythematosus and its effect / 上海交通大学学报（医学版）

Objective: To study the expression of transcription factor BTB and CNC homology 2 (Bach2) in CD4+CD25+CD45RA- T cells from patients with systemic lupus erythematosus (SLE) and its effect on cell function. Methods: The CD4+CD25+CD45RA- T cells from active SLE patients and healthy volunteers were sorted by flow cytometry. The expression of Bach2 in CD4+CD25+CD45RA- T cells was detected by fluorescence quantitative PCR and Western blotting. The correlation between the median flourscence indensity (MFI) of Bach2 in CD4+CD25+CD45RA- T cells and the disease activity index of SLE (SLEDAI) was analyzed. The MFI of Bach2 in IL-17+CD4+CD25+CD45RA- T cells was compared with that in IL-17-CD4+CD25+CD45RA- T cells by flow cytometry. In Bach2 overexpression system, the expression of IL-17 in CD4+CD25+CD45RA- T cells was detected by flow cytometry and the concentration of IL-17 in the culture supernants was detected by ELISA. Results: The mRNA and protein expressions of Bach2 in CD4+CD25+CD45RA- T cells from SLE patients were significantly lower than those in healthy controls (P<0.01). There was a significant negative correlation between the MFI of Bach2 and SLEDAI (R2=0.433, P=0.001) in patients with SLE. The expression of Bach2 in IL-17+CD4+CD25+CD45RA- T cells was significantly lower than that in IL-17-CD4+CD25+CD45RA- T cells (P=0.013). When Bach2 was overexpressed, the percentage of CD4+CD25+CD45RA- T cells from SLE patients expressing inflammatory factor IL-17 decreased significantly (P=0.032) and the IL-17 concentration in cell culture supernatants markedly decreased (P=0.008). Conclusion: The expression of Bach2 in CD4+CD25+CD45RA- T cells from SLE patients decreases, and overexpression of Bach2 in the cells leads to the falling expression of IL-17.

Expression of transcription factor Bach2 in CD4+CD25+CD45RA-T cells in patients with systemic lupus erythematosus and its effect / 上海交通大学学报（医学版）

Objective·To study the expression of transcription factor BTB and CNC homology 2 (Bach2) in CD4+CD25+CD45RA-T ceils from patients with systemic lupus erythematosus (SLE) and its effect on cell function.Methods·The CD4+CD25+CD45RA T cells from active SLE patients and healthy volunteers were sorted by flow cytometry.The expression of Bach2 in CD4+CD25+CD45RA T cells was detected by fluorescence quantitative PCR and Western blotting.The correlation between the median flourscence indensity (MFI) of Bach2 in CD4+CD25+CD45RA-T cells and the disease activity index of SLE (SLEDAI) was analyzed.The MFI of Bach2 in IL-17+CD4+CD25+CD45RA-T ceils was compared with that in IL-17-CD4+CD25+CD45RA-T cells by flow cytometry.In Bach2 overexpression system,the expression of IL-17 in CD4+CD25+CD45RA T ceils was detected by flow cytometry and the concentration of IL-17 in the culture supernants was detected by ELISA.Results·The mRNA and protein expressions of Bach2 in CD4+CD25+CD45RA-T cells from SLE patients were significantly lower than those in healthy controls (P＜0.01).There was a significant negative correlation between the MFI of Bach2 and SLEDAI (R2=0.433,P=-0.001) in patients with SLE.The expression of Bach2 in IL-17+CD4+CD25+CD45RA-T cells was significantly lower than that in IL-17-CD4+CD25+CD45RA-T cells (P=-0.013).When Bach2 was overexpressed,the percentage of CD4+CD25+CD45RA-T cells from SLE patients expressing inflammatory factor IL-17 decreased significantly (P=0.032) and the IL-17 concentration in cell culture supernatants markedly decreased (P=0.008).Conclusion·The expression of Bach2 in CD4+CD25+CD45RA-T cells from SLE patients decreases,and overexpression of Bach2 in the cells leads to the falling expression of IL-17.

The effect of BTB and cap'n'collar proteinhomology 1 siRNA on the expression of antioxidant factors of lung fibroblasts in transforming growth factor-β1-induced interstitial fibrosis in mouse / 中华风湿病学杂志

Objective To investigate the effects of adenovirus vectors for BTB and cap'n'collar protein homology 1 (Bach1) small interfering RNA (siRNA) on antioxidanffactors and fibrosis related cytokines in lung fibroblasts (MLF) in transforming growth factor (TGF)-β1 induced mouse.Methods Bach1 siRNA recombinant adenovirus vectors and blankadenovirus vectorwere constructed,then the MLF cells were incubated with TGF-β1 (5 ng/ml) for 24 h and infected with blankvector and successful constructed Bach1 siRNAs.The messenger RNA (mRNA) and protein expression of Bach1,heme oxygenase (HO)-1,glutathione peroxidase 1 (GPx1) and nicotinamide-adenine dinucleotide phosphate:quinone oxidoreductase-1 (NQO1) in cell supernatants were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.Changes of fibrosis-related cytokines including TGF-β1 and interleukin (IL)-6 levels in cell supernatants were determined by enzyme-linked immunosorbent assay (ELISA).One-way analysis of variance (ANOVA) was performed for multiple group comparisons and LSD test was used to compare the two groups.Results Bach1 mRNA (2.127±0.089) and protein expression increased significantly after TGF-31 stimulation compared with blank group (1.000±0.067,t=-21.77,P＜0.01),as well as the expression of fibrosis-related cytokines TGF-β1 (52±6) and IL-6 (34±6) in cell supernatants increased significantly after TGF-β1 stimulation compared with blank group (26±4,t=-11.11,P＜0.01 and 20±5,t=-5.32,P＜0.01),but the mRNA expression of HO-1 and GPx1 (0.403±0.040 and 0.567±0.112) also the protein expression decreased significantly compared with mRNA (1.000±0.181,t=25.57,P＜0.01 and 1.000±0.212,t=6.68,P＜0.05) and protein expression in blank group.Follow the Bach1 siRNA treatment,Bach1 mRNA (0.153±0.015) and protein levels were significantly downregulated compared with mRNA (2.129±0.089 and 1.973±0.035,F=1835.95,P＜0.01) and protein expression of TGF-β1 and blank vector group,as well as TGF-β1 (26±3) and IL-6 (11±3) expression in cell supernatant were significantly inhibited compared with TGF-31 (52±6 and 34±6) and blankvector group (49±5 and 33±6) (F=22.25,P＜0.01 and F=28.38,P＜0.01).But the mRNA levels of HO-1 (3.303±0.294) and GPx1 (1.840±0.231) in MLF were promoted significantly compared with TGF-β1 (0.403±0.040 and 0.567±0.112) and blank vector group (0.353±0.057 and 0.667±0.090) (F=53.90,P＜0.01 and F=526.25,P＜0.01).Conclusion Silencing Bach1 rescues TGF-β1 induced reduction of antioxidants and increasethefibrosis in MLF cells.The study offers an experimental basis to explore pathogenesis of oxidative stress and antioxidant therapy for connective tissue disease related inter-stitial lung disease.

BPOZ inhibits proliferation of breast cancer cell lines / 军事医学

Objective To investigate the effect of BTB/POZ ( broad complex, tramtrack and bric a brac/poxviruses and zinc finger) on proliferation of breast cancer cell lines.Methods The eukaryotic expression plasmid pCMV-HA-BPOZ was constructed by cloning from cDNA of human genome.Western blotting was used to detect the expression of pCMV-HA-BPOZ.Cells growth assay was used to detect the effect of BPOZ on proliferation and colony formation assay was used to detect the effect of BPOZ on cell growth ability.Results Western blotting showed that HA-BPOZ was efficiently expressed in cells.Moreover, the growth ability and proliferation of cells were significantly inhibited in BPOZ overexpressed cells compared with the control cells (both P <0.05).Conclusionp CMV-HA-BPOZ plasmid is constructed.BPOZ can restrain breast cancer cell lines MCF7 and ZR-75-1 cells from proliferating and growing.The results of our study can con-tribute to the study of functions of BPOZ in breast cancer.

Application of Bromocresol Green and Bromothymol Blue for the Extractive Spectrophotometric Determination of Anti-hypertensive Drugs.

Simple, selective and highly sensitive spectrophotometric methods are proposed for the rapid and accurate determination of anti-hypertensive drugs namely telmisartan (TEL), propranolol (PRO), bisoprolol (BIS) and carvedilol (CRV) in tablets and biological fluids using bromocressol green (BCG) and bromothymol blue (BTB). The developed methods involve formation of stable yellow colored dichloromethane extractable ion-pair complexes of the amino derivative of four antihypertensive drugs such as TEL, PRO, BIS and CRV with two sulphonphthalein acid dyes, namely; BCG and BTB in acidic buffer. The effect of optimum conditions via pH on the ion-pair formation, reagent concentration, time and temperature and solvent was studied. The composition of the ion-pairs was found 1: 1 by Job’s method. The established methods having high sensitivity and good selectivity could be applied to the determination of the studied drugs in pharmaceutical, urine and blood serum samples with satisfactory results. The results obtained are good agreement with experimental data. The reaction mechanism was also discussed.

Discovery of direct inhibitors of Keap1-Nrf2 protein-protein interaction as potential therapeutic and preventive agents

The Keap1-Nrf2-ARE pathway is an important antioxidant defense mechanism that protects cells from oxidative stress and the Keap1-Nrf2 protein-protein interaction (PPI) has become an important drug target to upregulate the expression of ARE-controlled cytoprotective oxidative stress response enzymes in the development of therapeutic and preventive agents for a number of diseases and conditions. However, most known Nrf2 activators/ARE inducers are indirect inhibitors of Keap1-Nrf2 PPI and they are electrophilic species that act by modifying the sulfhydryl groups of Keap1׳s cysteine residues. The electrophilicity of these indirect inhibitors may cause "off-target" side effects by reacting with cysteine residues of other important cellular proteins. Efforts have recently been focused on the development of direct inhibitors of Keap1-Nrf2 PPI. This article reviews these recent research efforts including the development of high throughput screening assays, the discovery of peptide and small molecule direct inhibitors, and the biophysical characterization of the binding of these inhibitors to the target Keap1 Kelch domain protein. These non-covalent direct inhibitors of Keap1-Nrf2 PPI could potentially be developed into effective therapeutic or preventive agents for a variety of diseases and conditions.

Suppression of hepatocellular carcinoma invasion and metastasis by Rho-kinase inhibitor Fasudil through inhibition of BTBD7-ROCK2 signaling pathway / 中南大学学报(医学版)

Objective: To explore the eff ect of Fasudil on the invasion and metastatic abilities of human high metastatic liver cancer cells (HCCLM3) and the underlying mechanisms. Methods: HCCLM3 cells were incubated with 100 μmol/L Fasudil. Fluorescence staining forF-actin and Transwell assay were performed to observe the invasion ability of HCCLM3 cells. HCCLM3 cells were divided into 3 groups: a negative control group, a Fasudil group and a BTB/POZ domain containing 7 (BTBD7)-siRNA group. Western blot assay was performed to detect the expression levels of BTBD7, ras homolog family member C (RhoC) and Rho-associated, coiled-coil containing protein kinase 2 (ROCK2), matrix metalloproteinases 2 (MMP2) and MMP9. Zymogram analysis method was performed to detect the expression activities of MMP2 and MMP9. hTe BTBD7-siRNA group was served as a positive control. Results: In HCCLM3 cells treated with Fasudil, the invasion ability was significant decreased compared with the control group, concomitant with the down-regulated expression levels of BTBD7, RhoC and ROCK2 protein as well as the decreased activities of MMP2 and MMP9. Conclusion: Fasudil plays an important role in interfering BTBD7-ROCK2 signaling pathway and suppressing the invasion and metastasis of hepatocellular carcinoma.

Rotary Instability after Anterior Cruciate Ligament Reconstruction with Single BTB Tendon / 대한정형외과학회잡지

PURPOSE: To analyze both the functional restoration and recovery of rotational and anterior-posterior stability after a single bundle ACL reconstruction using a BTB tendon. MATERIALS AND METHODS: A total of 52 patients were evaluated with an average follow up period of 32 months. A Lachman test, KT-2000 arthrometer, and Pivot shift test were performed to analyze the AP and rotational stability of the patients. The IKDC and Lysholm score was then used to evaluate the clinical results of the patients. The correlation between femoral tunnel angle and recovery of rotational stability was evaluated to determine the association between the two variables. This study also evaluated how the recovery of rotational stability affects the functional recovery of the patients. RESULTS: At the final follow up, the results indicated significant improvement according to the negative Lachman tests in 40 cases (76%), with an average of 2.92 mm anterior translation in the KT-2000 arthrometer and negative Pivot shift tests in 41 cases (79%)(p<0.05). The Lysholm and IKDC scores also showed significant improvement (p<0.05). Throughout the study, Group A was designated as those with <5 mm anterior translation and a negative Pivot shift test whereas Group B had positive test results. In Group A, the results showed 35 normal (85%), and 6 near normal (15%) cases in the IKDC score system, whereas Group B showed 2 normal (25%) and 5 near normal (62.5%) cases. Group A had an average of 89.3 in the Lysholm score system whereas Group B had a score of 60.5. On the knee tunnel view, Group A showed an average femoral tunnel angle of 49.2degrees, whereas Group B showed 63.5degrees. CONCLUSION: Decreasing the inclination of the BTB tendon using a transtibial femoral tunnel angle at either 10'30 or 1'30 will result in an excellent clinical outcome by achieving both anterior and rotational stability when operating a single bundle ACL reconstruction.

MORPHOLOGICAL OBSERVATION IN VITRO MODEL OF BLOOD-TUMOR BARRIER OF GLIOMA / 解剖学报

Objective To study the morphological characteristics of blood-tumor barrier(BTB)model of glioma in vitro. Methods After C 6 glioma cells/endothelia ECV 304 co-cultured mixed or in Transwell or on both sides of membrane of Transwell,the morphological characters of fenestra of endothelial cells,the junction between ECV 304 cells,the interface between C 6 cells and ECV 304 cells,and perivascular-end-feet of C 6 cells were observed under transmission electron microscope(TEM)and scanning electron microscope(SEM),and compared to BTB in 4 cases with human brain glioma. Results ECV 304 cells grown to confluence were not fenestrated,but with fomation of tight junction between ECV 304 cells after co-cultured with C 6 cells mixedly,in Transwell or on both sides of membrane of Transwell;It was not found that pseudopodia from C 6 cells as co-cultured in Transwell reaching into pore of Transwell;C 6 cells co-cultured on both sides of membrane of Transwell often sticked out to the ECV 304 cells,but with no pseudopodia from C 6 cells surrounded ECV 304 cells of penetrated the endothelia clefts.Perivascular-end-feet of C 6 cells were not integrant.These characters were similar to BTB in human brain glioma.Conclusion C 6 glioma cells/endothelia cell ECV 304 co-cultures on both sides of membrane of Transwell may simulate the morphological characters of BTB in vivo in some degree.;