Effect of CKIP-1 on hepatocyte apoptosis in nonalcoholic fatty liver disease / 中华内科杂志

Objective:To explore the effect and underlying mechanism of casein kinase 2 interacting protein-1 (CKIP-1) on hepatocyte apoptosis in nonalcoholic fatty liver disease (NAFLD).Methods:Experimental study. An NAFLD cell model was established by inducing human hepatoma cell line, HepG 2 cells, with oleic acid (OA). Flag-CKIP-1 expression vector and shRNA-CKIP-1 were transfected into HepG 2 cells. Flow cytometry was used to detect the effect of CKIP-1 on the activity and apoptosis of NAFLD hepatocytes. The levels of apoptosis-related proteins were detected by Western blot. CKIP-1 knockout mice in C57BL/6 back-ground were fed with either standard or high-fat diet for 8 weeks. Apoptosis-related signal proteins in NAFLD hepatocytes were detected by immunohistochemistry. Results:After CKIP-1 was transfected into HepG 2 cells, the degree of OA induced cell liposis was significantly reduced ( P<0.05). Annexin V-FITC/PI flow cytometry showed that CKIP-1 reduced the apoptosis of steatotic hepatocytes. Overexpression of CKIP-1 could significantly inhibit the expression of caspase-3 and caspase-9 and increase the expression of Bcl-2/Bax ( P<0.05). Knockdown of CKIP-1 could increase the expression of caspase-3 and caspase-9 ( P<0.05). CKIP-1 knockout could further increase the expression of caspase-3 and caspase-9 in NAFLD mice ( P<0.01, P<0.05), and further decrease the expression of Bcl-2/Bax ( P<0.05). Conclusion:CKIP-1 inhibited the apoptosis of steatotic hepatocytes by up-regulating the expression of apoptosis inhibitor gene, Bcl-2/Bax, and affecting the proteases, caspase-3 and caspase-9.

In vitro release mechanism and cytotoxic behavior of curcumin loaded casein nanoparticles

Abstract In the recent past, drug delivery through nanoparticles is considered an effective tool to treat various diseases. Biopolymeric nanoparticles such as protein based nanoparticles have vital role as drug carrier as it is non-antigenic, and easily biodegradable. Curcumin, plant polyphenolic anticancerous compound was loaded into the casein nanoparticles by coacervation method. Particle size and surface charge of spherical casein nanoparticles as observed to be 201.4 nm and -86.9 mV. The loading efficiency of curcumin loaded casein nanoparticles was found to 85.05 %. In vitro drug release was performed at different pH (7.4 and 3.0), and the cumulative release was observed to be 24.8 and 20.13% and at different temperatures (25°C and 37°C), the cumulative release was observed to be 24.8 and 28.60 % respectively in 48 h. Curcumin release from casein nanoparticles was shown to be in a steady, and prolonged rate. The nanoparticles were observed to have an effective antimocrobial activity than curcumin in free form. The drug loaded casein nanoparticles were found to be potent particles to protect cells from hydrogen peroxide and UV light damage. The cytotoxic activity of nanoparticles on MCF7 and A549 cells were assayed and was observed to have an IC50 value of 609 and 825.2µg/ml. Cell death was observed to be through apoptosis, accompanied by DNA fragmentation.

Research progress on natural antibacterial ingredients of glass ionomer cement / 口腔疾病防治

@#Glass ionomer cement (GIC) is widely used as a common filling material in dentistry but still exhibits problems with secondary caries and fractures. Thus, the antibacterial and anti-caries performance of GIC needs to be further improved. In recent years, natural antimicrobial components have become more desirable due to their good biological properties and low drug resistance. In this review, the natural antimicrobial ingredients in GIC modification are classified, reviewed and summarized according to the different sources of antimicrobial ingredients. In terms of animal origin, chitosan and casein phosphopeptide-amorphous calcium phosphate exhibit antimicrobial properties without affecting the mechanical properties of materials; propolis and bioactive enzymes have good biocompatibility; in terms of plant origin, polyphenols help improve the antimicrobial and mechanical properties of the material; arginine has a good remineralization effect; and plant essential oils have a certain ion release effect. In terms of microbial origin, antibiotics greatly improve the antibacterial properties of materials; in addition, the combined application of natural antimicrobial ingredients also exhibited excellent performance. Despite these advantages, the optimal addition concentration and biocompatibility in vivo are questions that need to be further explored before clinical applications can be achieved.

WSB1 regulates c-Myc expression through β-catenin signaling and forms a feedforward circuit

The dysregulation of transcription factors is widely associated with tumorigenesis. As the most well-defined transcription factor in multiple types of cancer, c-Myc can transform cells by transactivating various downstream genes. Given that there is no effective way to directly inhibit c-Myc, c-Myc targeting strategies hold great potential for cancer therapy. In this study, we found that WSB1, which has a highly positive correlation with c-Myc in 10 cancer cell lines and clinical samples, is a direct target gene of c-Myc, and can positively regulate c-Myc expression, which forms a feedforward circuit promoting cancer development. RNA sequencing results from Bel-7402 cells confirmed that WSB1 promoted c-Myc expression through the β-catenin pathway. Mechanistically, WSB1 affected β-catenin destruction complex-PPP2CA assembly and E3 ubiquitin ligase adaptor β-TRCP recruitment, which inhibited the ubiquitination of β-catenin and transactivated c-Myc. Of interest, the effect of WSB1 on c-Myc was independent of its E3 ligase activity. Moreover, overexpressing WSB1 in the Bel-7402 xenograft model could further strengthen the tumor-driven effect of c-Myc overexpression. Thus, our findings revealed a novel mechanism involved in tumorigenesis in which the WSB1/c-Myc feedforward circuit played an essential role, highlighting a potential c-Myc intervention strategy in cancer treatment.

Estructura, propiedades y genética de las caseínas de la leche: una revisión / Structure, properties and genetic of milk caseins: a review / Estrutura, propriedades e genética das caseínas do leite: uma revisão

Resumen La leche es un alimento esencial para los humanos y una de sus importancias radica en el contenido de proteínas lácteas. Las proteínas más frecuentes en este preciado líquido son las caseínas (αS1-caseína, αS2-caseína, β-caseína y κ-caseína), las cuales son fuente de aminoácidos para la dieta de los mamíferos en sus primeros días de vida. En la leche, las caseínas, están formadas por agregados moleculares de proteínas de tamaños variables denominados micelas. El objetivo de esta revisión es presentar un panorama general de la estructura, propiedades y genética de las caseínas lácteas y su relación con la salud humana. A partir de esta revisión, se pudo establecer, que las αs1 y αs2 caseínas se encuentran en conjunto con la β-caseína, formando el núcleo micelar, interactuando con los iones de calcio, para formar y mantener la micela estable. Animales caracterizados genéticamente con algunas variantes de estas proteínas, se asocian con un rendimiento en el volumen de leche. La κ-caseína, por su parte, está asociada con un aumento en el rendimiento y calidad de los quesos, de ahí su importancia económica, mientras que las formas más comunes de β-caseína en razas de ganado lechero son A1 y A2. La β-caseína A2 no presenta efectos negativos a la salud humana, por el contrario, ha sido asociada con propiedades reductoras de colesterol y triacilglicéridos. Sin embargo, la variante A1 de la β-caseína produce un péptido bioactivo denominado β-casomorfina-7 (BCM-7), que puede desempeñar un papel etiológico poco claro en el desarrollo de algunas enfermedades en humanos, tales como: enfermedad isquémica del corazón, diabetes mellitus tipo 1, síndrome de muerte súbita infantil (SIDS), desórdenes neurológicos, como autismo y esquizofrenia.

Abstract Milk is an essential food for humans and one of the reasons of its importance lies in the content of milk proteins. The most frequent proteins in this precious liquid are caseins (αS1-casein, αS2-casein, β-casein and κ-casein), which are a source of amino acids for the diet of mammals in their first days of life. In milk, caseins are made up of molecular aggregates of proteins of varying sizes called micelles. The objective of this review is to present an overview of the structure, properties and genetics of dairy caseins and their relation with human health. From this review, it was established that αs1 and αs2 caseins are found together with β-casein, forming the micellar nucleus and interacting with calcium ions, to form and maintain stable the micelle. Animals genetically characterized with some variants of these proteins are associated with a yield in milk volume. For its part, κ-casein is associated with an increase in the yield and quality of cheeses, hence its economic importance, while the most common forms of β-casein in dairy cattle are A1 and A2. β-casein A2 does not have negative effects on human health; on the contrary, it has been associated with lowering properties of cholesterol and triacylglycerides. However, the A1 variant of β-casein produces a bioactive peptide called β-casomorphin-7 (BCM-7), which may play an unclear etiological role in the development of some diseases in humans, such as: ischemic heart disease, type 1 diabetes mellitus, sudden infant death syndrome (SIDS), neurological disorders, such as autism and schizophrenia.

Resumo O leite é um alimento essencial para o ser humano e uma de suas principais característica é o teor de proteínas do leite. As proteínas mais frequentes neste líquido são as caseínas (αS1-caseína, αS2-caseína, β-caseína e κ-caseína), que são uma fonte de aminoácidos para a dieta dos mamíferos nos primeiros dias de vida. As caseínas no leite são constituídas por agregados moleculares de proteínas de variados tamanhos, chamados micelas. O objetivo desta revisão é apresentar uma visão geral da estrutura, propriedades e genética das caseínas lácteas e sua relação com a saúde humana. A partir desta revisão, foi estabelecido que as caseínas αs1 e αs2 são encontradas em conjunto com a β-caseína, formando o núcleo micelar, interagindo com os íons cálcio, para formar e manter a micela estável. Animais geneticamente caracterizados com algumas variantes dessas proteínas estão associados com o rendimento da produção de leite. Por sua vez, a κ-caseína está associada ao aumento do rendimento da produção e da qualidade dos queijos, por isso sua importância econômica, enquanto as formas mais comuns de β-caseína em bovinos leiteiros são A1 e A2. A Β-caseína A2 não tem efeitos negativos na saúde humana, pelo contrário, tem sido associada a propriedades redutoras do colesterol e dos triacilglicéridos. No entanto, a variante A1 da β-caseína produz um peptídeo bioativo denominado β-casomorfina-7 (BCM-7), que pode desempenhar uma função etiológico ainda desconhecida no desenvolvimento de algumas doenças em humanos, tais como: doença isquêmica do coração, diabetes mellitus tipo 1, síndrome da morte súbita infantil (SMSL), distúrbios neurológicos, como autismo e esquizofrenia.

Effect of preconditioning the fractured coronal fragments with remineralizing agents on fracture resistance of re-attached teeth

Background: Fragment re-attachment has been considered as one of the treatment modalities for the management of fractured anterior teeth. Hydration of fractured fragments aids in inhibiting the loss of ions and maintains vitality and esthetics. Aim: The study aimed to evaluate the effect of preconditioning the fractured fragments with remineralizing agents on fracture resistance of re-attached teeth. Settings and Design: This was an in vitro study. Materials and Methods: Sixty freshly extracted noncarious human permanent maxillary central and lateral incisors were randomly allocated into three Groups of 20 each: Group 1: 2% sodium fluoride (2%NaF), Group 2: casein phosphopeptide–amorphous calcium phosphate (CPP-ACP), and Group 3: self-assembling peptide P11-4 (SAP). These were further divided into two subgroups of 10 teeth each, based on contact time with remineralizing agents, i.e., 30 min and 2 h. Fractured fragments were treated with remineralizing agents for a specified contact time and then re-attached with flowable composite resin. Force required to fracture the re-attached tooth was recorded in Newtons using universal testing machine. Statistical Analysis: Unpaired t-test, one-way analysis of variance test, and post hoc Tukey test were used for the statistical analysis. Results: A higher fracture resistance was noticed in fragments treated with 2% NaF (30 min- 215.6 N, 2 h- 188.5 N) compared to CPP-ACP (30 min- 141.3 N, 2 h- 111.1 N) and SAP (30 min- 134.8 N, 2 h- 149.5 N). At 30 min interval, it was found to be more in 2% NaF and CPP-ACP groups compared to 2 h. However, it increased with time in the SAP group. A statistically significant difference was found between the groups at both time intervals (P = 0.007 and 0.017). Conclusion: Preconditioning of fractured coronal fragments with 2% NaF showed higher fracture resistance compared to CPP-ACP and self-assembling peptide P11-4. Samples treated with SAP P11-4 exhibited good fracture resistance at 2 h contact time.

Análisis genético de cinco polimorfismos de nucleótido simple de caseínas lácteas obtenidos con chips genómicos en ganado Holstein de Antioquia, Colombia / Genetic analysis of five single nucleotide polymorphisms of dairy caseins obtained with genomic chips in Holstein cattle from Antioquia, Colombia

RESUMEN Los polimorfismos genéticos asociados con las caseínas de la leche son de gran importancia, ya que pueden ser usados como marcadores genéticos para mejorar el rendimiento productivo en los hatos lecheros. El objetivo de este estudio fue evaluar la diversidad y estructura genética de 5 SNP de caseínas de la leche, obtenidos con chips genómicos en vacas y toros de raza Holstein en Antioquia (Colombia). Fueron muestreados 113 animales de raza Holstein en 3 regiones del departamento de Antioquia (norte, centro y oriente) y un cuarto grupo de sementales comerciales. Los animales fueron genotipificados con chips genómicos de alta densidad (Illumina BovineHD e Illumina SNP50 v2), a partir de los cuales se identificaron 5 SNP (ARS-BFGL-NGS-8140, BTA-77380-no-rs, BTA-32346-no-rs, BTB-00821654 y ARS-BFGL-NGS-15809). Para cada SNP se realizó un análisis genético mediante un análisis de varianza molecular (amova) usando el software GenAIEx 6.501. Los SNP con mayor heterocigosidad total (HT) fueron ARS-BFGL-NGS-8140 y BTA-32346-no-rs, con resultados cercanos al 45%; sin embargo, la Ht para ARS-BFGL-NGS-15809, BTA-77380-no-rs y BTB-00821654 estuvo por debajo del 15%. El SNP con mayor diversidad genética fue BTA-32346-no-rs (Ho-He = 0,06; p < 0,05). En esta investigación se evaluó una subpoblación de toros comerciales extranjeros, en la cual se obtuvieron frecuencias alélicas y genotípicas similares a las obtenidas para las subpoblaciones locales, sugiriendo que los alelos de los toros muy posiblemente están fijados en dichas subpoblaciones, por lo que la estructura y diversidad genética tienden a ser bajas en la muestra de estudio.

ABSTRACT Genetic polymorphisms associated with milk caseins have a great importance since they can be used as genetic markers to improve productive performance in dairy herds. The main goal of the present study was to evaluate the diversity and genetic structure of 5 SNPs of milk caseins, obtained with genomic chip in Holstein cows and bulls from Antioquia (Colombia). 113 Holstein animals were sampled in 3 regions of Antioquia (north, center, and east), and a fourth group of commercial sires. Animals were geno-typed with high-density SNP chips (Illumina BovineHD and Illumina SNP50 v2), from which 5 SNPs were identified (ARS-BFGL-NGS-8140, BTA-77380-no-rs, BTA-32346-no-rs, BTB-00821654 and ARS-BFGL-NGS-15809). For each SNP, a genetic analysis was performed by means of an analysis of molecular variance (AMOVA) using the GenAIEx 6.501 software. The SNPs with the highest total heterozygosity (Ht) were ARS-BFGL-NGS-8140 and BTA-32346-no-rs, with results close to 45%; however, the HT for ARS-BFGL-NGS-15809, BTA-77380-no-rs, and BTB-00821654 were below 15%. The SNP with the highest genetic diversity was BTA-32346-no-rs (Ho-He = 0,06; p < 0,05). In this research a subpopulation of foreign commercial bulls was evaluated, in which similar allelic and genotypic frequencies to those for local subpopulations were obtained, suggesting that the alleles of the bulls are very possibly fixed in these subpopulations, so that the structure and genetic diversity tend to be low in the study sample.

Validation of a method employing liquid chromatography to identify and quantify glycomacropeptide of casein in raw milk / Validación de un método mediante cromatografía líquida para identificar y cuantificar glicomacropéptido de caseína en la leche cruda

Background: The information on official testing methods, or regulatory methods in Colombia to test whey in milk is limited; this restriction of information goes against the possibility of mitigating the risk of food fraud. Objectives: The validation of an HPLC method to determine casein glycomacropeptide (c-GMP), a protein that countries such as Brazil, Spain, and Ecuador have used as an indicator of raw milk adulteration with whey, was carried out. Methods: A 10mL sample of raw milk is precipitated with 24% TCA using ultrasound, a process followed by filtration. The collected fraction ensured the separation of c-GMP and then injected into the liquid chromatography. Results: A 30 minutes analysis allowed the determination of c-GMP with a retention time of 12.9 ± 0.5 minutes. The performance characteristics method in the validation exercise were: recovery percentage 99.97%, linearity R2> 0.95; % RSD accuracy <5.3%. Conclusion, the method exhibits desirable attributes for the intended purpose

Antecedentes: En Colombia la información de dominio público en metodologías de análisis de lactosuero en leche es limitada, restringiendo la posibilidad de acceder a ellas para mitigar el riesgo de fraude alimentario. Objetivos: Se realizó validación de un método por HPLC para determinar en leche cruda c-GMP, proteína usada como indicador de adulteración en países como Brasil y Ecuador. Metodos: Una muestra de 10mL de leche cruda es precipitada con TCA al 24% empleando ultrasonido, proceso seguido por filtración. La fracción recolectada aseguró la separación del c-GMP para luego inyectar al cromatógrafo líquido. Resultados: La determinación de c-GMP permitió el análisis en 30 minutos con tiempo de retención de 12,9 ± 0,5 minutos. Las características de desempeño del método en el ejercicio de validación fueron: porcentaje de recuperación 99,97%, linealidad R2>0,95; precisión %RSD< 5,3%. Conclusión: el método al final del ejercicio exhibe atributos para el fin previsto

Allele and genotype frequency for milk beta-casein in dairy cattle in the northern region of Tocantins State, Brazil / Frequências alélicas e genotípicas para beta-caseína do leite em bovinos leiteiros da microrregião de Araguaína, Tocantins

At present, there is a concern about the quality of milk and diseases related to its consumption, as it can generate discomfort and allergic reactions in some individuals due to its protein components. Thus, the present study was developed to identify the allele and genotype frequencies of genes for ß-casein, A1 and A2, in dairy herds in the region of Araguaína-TO, Brazil. Genetic material from 421 animals (crossbred dairy cattle in lactation) was used. All animals were numbered for identification, and DNA samples were extracted from hair bulbs. Samples for two markers from the polymorphic regions were characterized and confirmed by real-time PCR using the ABI Prism® 7500 Sequence Detection System (Applied Biosystems). Allele and genotype frequencies were determined using the TaqMan™ detection system, where the primer and probe release different fluorescence signals for each allele of the polymorphism. The sampled herd showed frequencies of 28.27% for the A1 allele and 71.73% for the A2 allele. Genotype frequencies were 52.96% (223/421) for A2A2; 37.53% (158/421) for the A1A2 genotype; and 9.50% (40/421) for the A1A1 genotype. The frequency of the A1 allele for ß-casein in dairy herds from the northern region of Tocantins was low and is per the results of previous studies. Although the A2A2 genotype of ß-casein had a high relative frequency, the A1A2 genotype is still rather frequent, warranting greater selection pressure.(AU)

Atualmente existe uma preocupação em relação à qualidade e doenças que estão relacionadas ao consumo de leite, pois o mesmo pode gerar desconfortos e reações alergicas em alguns indivíduos devido aos seus constituintes protéicos. Assim, o presente estudo teve como objetivo identificar a frequência alélica e genotípica de genes para beta caseína, A1 e A2, em rebanhos leiteiros da região de Araguaína-TO. Foram utilizados material genético de 421 animais (bovinos leiteiros mestiços em lactação), e todos os animais foram numerados para identificação e amostras de DNA foram extraídas de bulbo de folículos pilosos. As amostras para dois marcadores das regiões polimórficas foram caracterizadas e confirmadas por PCR em tempo real, usando um sistema de detecção de sequências ABI Prism® 7500 (Applied Biosystems). As frequências alélicas e genotípicas foram determinadas utilizando o sistema de detecção TaqMan ™, no qual o primer e a sonda emitem diferentes sinais de fluorescência para cada alelo do polimorfismo. Observou-se frequência do alelo A1 de 28,27%, e do alelo A2 de 71,73% no rebanho amostral. A frequência genotípica de A2A2 foi de 52,96% (223/421), com genótipo A1A2 de 37,53% (158/421), e de 9,50% (40/421) animais com genótipo A1A1. A frequência do alelo A1 para beta-caseína em rebanhos leiteiros da região norte do Tocantins foi baixa e seguiu a mesma tendência já observada em estudos anteriores. Os genótipos A2A2 da beta-caseína apresentaram frequência relativa alta, entretanto o genótipo A1A2 ainda é bastante frequente, necessitando de maior pressão de seleção.(AU)

A novel technology to reduce astringency of tea polyphenols extract and its mechanism / 中草药·英文版

Objective: Tea polyphenols are natural extracts used widely throughout the world. However, the severe astringency of tea polyphenols has reduced patient compliance. Based on the analysis of the formation mechanism of astringency, this paper hopes to propose a new method to control the astringency of tea polyphenols and improve patient compliance without changing its effect. Methods: Artificial saliva was used to prepare the tea polyphenols solution with different pH, using β-casein to imitate salivary protein, and preparing 1.2 mg/mL β-casein solution. A fluorescence quenching test was used to study the interaction between tea polyphenols and β-casein, combined with the stability test results of the compound, we can choose the pH with weak binding but good stability as the best pH for masking astringency. The taste-masking tablets were prepared under the best pH conditions, and the Xinnaojian Original Tablets were prepared according to the conventional preparation method. The disintegration time limit and solubility were tested respectively. The astringency of Xinnaojian original tablets and taste-masking tablets was evaluated by visual analogue scale (VAS). Results: The result of the fluorescence quenching test prompted that the combination force was the weakest when the pH was 4.9. Further synchronous fluorescence analysis showed that an increase in pH resulted in a decrease of the binding sites between tea polyphenols and β-casein, and this decrease was closely related to changes in tryptophan residues in β-casein. Both original and taste-masking Xinnaojian Tablets were prepared. Volunteers’ VAS scores illustrated that the astringency improved significantly with the masking tablets (P < 0.05). Conclusion: This pH-adjusting masking treatment had little effect on the recovery of polyphenols from the tablets or the dissolution of the tablets. This study provides a novel and feasible astringency masking technology for tea polyphenols and its preparation.

Knowledge of Special Nutrition for Children with Autism Spectrum Disorder Among Parents and Special Educators in Malaysia (Pengetahuan Tentang Pemakanan Khas bagi Kanak-kanak yang Mengalami Gangguan Spektrum Autisme dalam Kalangan Ibu Bapa dan Pendidik Khas di Malaysia) / Jurnal Sains Kesihatan Malaysia

@#Sufficient intake of macronutrients and micronutrients are essential for child growth. Many children with autism spectrum disorder (ASD) however are reported to be picky eaters which may lead to malnourishment. As diet and behaviour are importantly interconnected, this study aims to determine the level of knowledge on nutrition among parents and special educators of children with ASD. A cross-sectional study was conducted at a National Autism Centre in Kuala Lumpur, Malaysia from May to August 2018. A 37-item questionnaire was self-administered to the respondents who attended series of training of trainers’ sessions. The data was the collected and analysed descriptively. A total of 209 respondents participated with 153 parents and 56 special educators of children with ASD. Only 37.9% of the parents agreed that their children’s diet contain balanced nutrition. Majority of the parents and special educators had the perception that sugar, junk food and gluten are bad for the children but they were still given due to the accessibility and convenience. The low level of knowledge among parents and special educators about the nutrition of children with ASD indicates that a measure should be taken to increase the awareness and to overcome the malnutrition among ASD children.

Expression and clinical correlation of casein kinase 1γ2 in head and neck squamous cell carcinoma / 国际生物医学工程杂志

Objective:To explore the role of casein kinase 1 gamma 2 (CSNK1G2) in the development and progression of head and neck squamous cell carcinoma (HNSC).Methods:Based on the Cancer Genome Atlas (TCGA), LinkedOmics and UALCAN were used to analyze the relationship among the mRNA expression of CSNK1G2, methylation, copy number variation and clinical indicators in HNSC, as well as to analysis CSNK1G2 related co-expression genes and proteins. The expression of CSNK1G2 in HNSC was verified by RT-qPCR experiments of clinical samples. Protein interaction network analysis on CSNK1G2 expression-related proteins was performed using STRING database.Results:UALCAN analysis showed that the expression of CSNK1G2 mRNA in HNSC was higher than that in normal tissues ( P<0.001), and the expression of CSNK1G2 mRNA was up-regulated in lower differentiation and Human Papilloma Virus (HPV)-positive HNSC (all P<0.05). But in HNSC with different pathological stages, different age stages and different lymph node metastasis stages (N stage), there was no difference in the amount of CSNK1G2 mRNA expression (all P>0.05). The RT-qPCR experiment confirmed the increased expression of CSNK1G2 mRNA in HNSC. LinkedOmincs analysis results showed that CSNK1G2 mRNA expression was positively correlated with CSNK1G2 copy number variation ( P<0.001) and negatively correlated with methylation ( P<0.001). Survival analysis results showed that high CSNK1G2 mRNA expression and copy number mutations predicted better survival ( P=0.033, P=0.015), while methylation levels were not associated with survival ( P=0.458). Gene set enrichment analysis results showed that CSNK1G2-related co-expression genes were mainly in DNA replication. The STRING's protein interaction network analysis results showed that TP53, CHEK1, and CHEK2 may be key proteins. These proteins are significantly associated with high expression levels of CSNK1G2. Conclusions:CSNK1G2 may cooperate with TP53, CHEK1 and CHEK2 related proteins to promote the development of HNSC and tumor proliferation, but does not affect the metastasis and spread of HNSC. An increase in the expression of CSNK1G2 in HNSC may indicate a better survival prognosis.

The effects of casein kinase 2 interacting protein-1 on the growth and development of craniomaxillofacial soft and hard tissues in mice / 口腔疾病防治

Objective@# To investigate the effect of casein kinase 2 interacting protein-1 (CKIP-1) on craniofacial soft tissues and hard tissues, to provide the basis for the study and treatment of craniomaxillofacial related diseases.@*Methods@#6-month- old male CKIP-1 knockout (KO) mice were selected as the experimental group, and wild-type (WT) mice were selected as the control group. The craniomaxillofacial hard tissues (parietal bone, nasal bone, incisors and molars) were analyzed through micro- CT, and the morphological changes of maxillofacial soft tissues (nasal cartilage, lip mucosa and tongue) were analyzed through HE staining and toluidine blue staining.@* Results@#CKIP-1 negatively regulated bone mass of cancellous bone of cranial and maxillofacial bones and dentin mineralization. Compared with the WT mice, the thickness of the parietal baffle layer increased by 93% in KO mice, while cortical bone showed no significant difference between the two groups. The nasal cancellous bone thickness increased by 160% in KO-mice, while cortical bone showed no significant difference between the two groups; the enamel thickness was normal, but the pulp cavity became smaller and the dentin thickness increased by 48%. Compared with the WT mice, the HE staining and toluidine blue staining analyses of the soft tissues revealed that the thickness of the alar cartilage plate of KO mice increased by 57%, and local ossification was found within the cartilage plate. The thickness of the keratinized layer of the labial mucosa increased by 170% in KO mice and the muscle fiber diameter of the lingual muscle increased by 45%. @*Conclusion@#CKIP-1 genes have different effects on the growth and development of various soft and hard tissues in the maxillofacial region of mice.

Targeting RAS phosphorylation in cancer therapy: Mechanisms and modulators

RAS, a member of the small GTPase family, functions as a binary switch by shifting between inactive GDP-loaded and active GTP-loaded state. RAS gain-of-function mutations are one of the leading causes in human oncogenesis, accounting for ∼19% of the global cancer burden. As a well-recognized target in malignancy, RAS has been intensively studied in the past decades. Despite the sustained efforts, many failures occurred in the earlier exploration and resulted in an 'undruggable' feature of RAS proteins. Phosphorylation at several residues has been recently determined as regulators for wild-type and mutated RAS proteins. Therefore, the development of RAS inhibitors directly targeting the RAS mutants or towards upstream regulatory kinases supplies a novel direction for tackling the anti-RAS difficulties. A better understanding of RAS phosphorylation can contribute to future therapeutic strategies. In this review, we comprehensively summarized the current advances in RAS phosphorylation and provided mechanistic insights into the signaling transduction of associated pathways. Importantly, the preclinical and clinical success in developing anti-RAS drugs targeting the upstream kinases and potential directions of harnessing allostery to target RAS phosphorylation sites were also discussed.

Advances in the structure and physiological function of protein kinase CK2 / 生物工程学报

Protein kinase CK2 is a common, evolutionarily conserved and ubiquitous protein kinase. In recent years, increasing evidences have shown that CK2 has a variety of phosphorylated protein substrates, which play important roles in growth, development and various diseases. Therefore, CK2 may participate in these physiological processes by regulating the phosphorylation of these substrates. This article briefly reviewed the structural characteristics of protein kinase CK2 and its physiological functions in growth, development, immunity, formation of tumor and other diseases, in order to provide knowledge basis for further research on the regulatory mechanism of CK2 and potential applications of its inhibitors.

Occluding dentin tubules with monetite paste / 华西口腔医学杂志

OBJECTIVES@#This study was performed to evaluate the occlusion of monetite paste on dentine tubule and provide a new potential method for treating dentine hypersensitivity.@*METHODS@#Calcium oxide, strontium chloride, and polyethylene glycol phosphate were mixed in a certain proportion and ground in a planetary ball mill. The reaction was carried out by adjusting the pH to obtain monetite and hydroxyapatite paste. The morphological characteristics of the paste were observed through scanning electron microscope (SEM). The structure and composition were analyzed through X-ray diffraction (XRD) and Fourier transform infrared spectrometer (FTIR). The extracted third molar was selected to undergo demineralization to establish the @*RESULTS@#XRD and FTIR showed that the composition of the paste was mainly monetite, and the composition of hydroxyapatite paste was mainly composed of hydroxyapatite. SEM revealed that the size of the crystal particles of the synthesized paste was tens to hundreds of nanometers. Monetite and hydroxyapatite paste could produce a thicker mineralization layer on the dentin surface, and the mineralization of the dentin tubules of monetite was deeper than that of hydroxyapatite paste. The microhardness of the monetite paste group was significantly less than those of the hydroxyapatite paste groups (@*CONCLUSIONS@#Monetite paste could effectively block the exposed dentin tubules and be used for treating dentin hypersensitivity.

Research progress in CPP-ACP combined with fluoride on enamel remineralization / 国际生物医学工程杂志

With the improvement of people's living standard and the strengthening of people's recognition of dental caries prevention and treatment, enamel remineralization has attracted extensive attention. Fluoride, bioactive glass and casein phosphopeptide-amorphic calcium phosphate (CPP-ACP) are the most widely studied remineralisation agents at present, which have a certain effect of promoting enamel remineralization. The results of different studies shown that the effect of CPP-ACP combined with fluoride was quite different, which may be affected by many factors. In this paper, the recent progress of the combined application of CPP-ACP and fluoride on enamel remineralization was reviewed, and various influencing factors were analyzed in order to provide references for future experimental design and clinical applications.

Influencia de la suplementación con vitamina D, omega 3 o el uso de una dieta libre de gluten y caseína en el comportamiento de niños y niñas con trastorno del espectro autista / Influence of the vitamin D, omega 3 supplementation or the use of a gluten and casein free diet in the behavior of boys and girls with autism spectrum disorder

La prevalencia de trastorno del espectro autista ha ido en aumento, sin embargo, en Chile no existen lineamientos nutricionales acerca del posible tratamiento de la condición. Es por ello que el objetivo de esta revisión fue analizar la evidencia actual en relación al uso de una dieta libre de gluten y caseína, suplementación de vitamina D y omega 3 y su impacto en el comportamiento de niños/as con trastorno del espectro autista. Hay evidencia con resultados en torno a los beneficios de la suplementación con vitamina D debido a su carácter neuroprotector y su función neuromuscular. A su vez, la evidencia con omega 3 (DHA) es estadísticamente significativa para irritabilidad, hiperactividad, letargo, comportamiento estereotipado, conciencia social, comunicación y disminución de la severidad del autismo. Respecto a la dieta libre de gluten y caseína lo observado es que no existe evidencia que respalde los beneficios que esta exclusión entregaría. Aún falta evidencia para declarar un manejo nutricional específico para el tratamiento de los síntomas gastrointestinales y de comportamiento, más allá de la suplementación con aquellos micronutrientes en déficit.

The prevalence rate of autism spectrum disorder has been increasing, however, in Chile there are no nutritional guidelines about the possible treatment of the condition. That is the reason why the aim of this review is to analyze the current evidence regarding the use of a gluten and casein free diet, vitamin D and omega 3 supplementation and its impact in the behavior of children's with spectrum disorder autistic. There is evidence based on significant results regarding the benefits of vitamin D supplementation due to its neuroprotective character and neuromuscular function. At the same time the omega 3 evidence is statistically significant in the diminution of irritability hyperactivity, lethargy, stereotypical behavior, severity of autism and increase of social consciousness and communication. Regarding the gluten-free and casein-free diet, what was observed is that there is no evidence to support the benefits that this exclusion would provide. The evidence has not been conclusive to declare a specific nutritional management for the treatment of gastrointestinal and behavioral symptoms, beyond supplementation with those micronutrients in deficit.

Immobilization of bromelain on cobalt-iron magnetic nanoparticles (CoFe2O4) for casein hydrolysis / Inmovilización de bromelina sobre nanopartículas magnéticas de cobalto-hierro (CoFe2O4) para la hidrólisis de caseína / Inmovilización de bromelina sobre nanopartículas magnéticas de cobalto-hierro (CoFe2O4) para a hidrólise de caseína

Abstract By means of recycling an enzyme, bromelain was used in casein hydrolysis facilitated by a nanobiocatalyst consisting of bromelain, CoFe2O4 magnetic nanoparticles, chitosan, and glutaraldehyde. Bromelain was immobilized on the chitosan cobalt-magnetite nanoparticle surface via covalent bonds to form the nanobiocatalyst. Immobilized bromelain showed 77% immobilization binding, retaining 85 ± 2% of the initial catalytic activity. Nanoparticles and immobilized bromelain were characterized using UV-Vis and IR spectroscopies, X-ray, square wave voltammetry (SWV), cyclic voltammetry (CV), vibrating-sample magnetization (VSM), and transmission electron microscope (TEM). The Michaelis-Menten constant (KM) and VMAX of the free and immobilized enzyme were calculated: KM = 2.1 ± 0.18 mM and 1.8 mM, respectively and VMAX = 6.08 x 10-2 ± 2.1 x 10-2 U/min and 6.46 ± 0.91 U/min, respectively. The thermal stability of the free enzyme was higher than the immobilized enzyme: 95-98% and 83-87%, respectively. An optimum pH of 6 and a temperature of 20 °C were determined in both cases. Immobilized bromelain maintained 50% of the initial catalytic activity after the fifth use. The immobilized bromelain proved to be effective and reusable for casein hydrolysis. As novel contribution the characterization by VOC and CV was carried out.

Resumen Se investigó la reutilización de la bromelina inmovilizada sobre nanopartículas magnéticas de CoFe2O4 para la hidrólisis de caseína. La inmovilización se efectuó covalentemente en un catalizador de nanopartículas de hierro y cobalto recubiertas con quitosano, glutaraldehído y bromelina. La bromelina alcanzó un máximo de inmovilización del 77%, conservando el 85 ± 2% de su actividad catalítica inicial a los 30 min. La caracterización del catalizador se realizó por espectrofotometría IR y UV-Vis, microscopía electrónica de transmisión (TEM), difracción de rayos X, voltametría de onda cuadrada (VOC), voltametría cíclica (VC) y magnetización de muestra vibrante (VSM). Los parámetros cinéticos KM y VMAX de la enzima libre e inmovilizada fueron: KM = 2,1 ± 0,18 mM y 1,8 ± 0,16 mM, respectivamente y VMAX = 6,08 x 10-2 ± 2,1 x 10-2 U/min y 6,46 ± 0,91 U/min, respectivamente. La estabilidad térmica de la enzima libre fue mayor que la de enzima inmovilizada: 95-98% y 83-87%, respectivamente. Se determinó un pH óptimo de 6 y temperatura de 20 °C en ambos casos. La bromelina inmovilizada mantuvo el 50% de su actividad catalítica hasta el quinto uso. Como aporte novedoso se realizó, en este estudio se realizó la caracterización por VOC y VC.

Resumo Foi estudado o reuso da enzima bromelina imobilizada sobre nano partículas magnéticas de CoFe2O4, para a hidrólise de caseína. A imobilização efetuou-se covalentemente em um catalisador de nano partículas de cobalto e ferro recobertas com quitosano, glutaraldehido e bromelina. A bromelina atingiu seu máximo de imobilização do 77% aos 30 minutos, conservando o 85 ± 2% da sua atividade catalítica inicial. A caraterização foi feita mediante espectrofotometria IV e UV-Vis, microscopia eletrônica de transmissão (TEM), difração de raios X, voltametria de onda quadrada (VOQ), voltametria cíclica (VC) e por magnetização de amostra vibrante (VSM). Os parâmetros cinéticos KM e VMAX da enzima livre e imobilizada foram KM = 2,1 ± 0,18 mM e 1,8 8 ± 0,16mM, respeitivamente; VMAX = 6,08 x 10-2 ± 2,1 x 10-2 U/min e 6,46 ± 0,91 U/min, respectivamente. A estabilidade térmica da enzima livre (95-98%) foi maior do que a estabilidade da enzima imobilizada (83-87%), determinou-se um pH óptimo de 6 e temperatura de 20 °C em ambos os casos. A bromelina imobilizada manteve o 50% de sua atividade catalítica até o quinto uso. Como aporte inovador neste estudo apresenta-se a caraterização por VOQ e VC.

Screening and identification of fibrinolytic enzyme-producing microbes in fermentation process of Sojae Semen Praeparatum / 中草药

Objective: To screen and identify the dominant strains which produce fibrinolytic enzyme during the processing of Sojae Semen Praeparatum (SSP, Dandouchi in Chinese). Methods: SSP was prepared according to the Chinese Pharmacopoeia (2020 edition), and samples were taken at different time points during the fermenting process of SSP.The casein plate method and fibrin plate method were used to screen the fibrinolytic enzyme-producing microorganisms in samples at different time points. The fibrinolytic enzyme-producing microorganisms were inoculated in the designated liquid medium to obtain single strain fermentation broth, and fibrin plate method was used to measure the fibrinolytic activity of the fermentation broth. The DNA sequences of fibrinolytic enzyme-producing bacteria and fungi were amplified using 16S rDNA and 18S rDNA universal primer by PCR respectively.The amplified products were sequenced, and the sequencing results were identified through NCBI homology comparison. Molecular biological identification was done by phylogenetic tree constructed by MEGA 4.1 software. Results: Three types of fibrinolytic enzyme-producing bacteria were screened out and identified in this study. They were Bacillus subtilis, Stenotrophomonas maltophilia and Micrococcus, respectively. The result of fibrin plate method showed that the fermentation broth of S. maltophilia had the highest fibrinolytic activity, reaching 527.49 IU/mL. Conclusion: There are fibrinolytic enzyme-producing dominant microorganisms existing in the fermenting process of SSP and the thrombolytic effect of SSP is worthy of further study. This study lays the foundation for revealing the formation mechanism of fibrinolytic enzyme in the fermentation process of SSP.