Effects of Siglec-E on parthenolide inhibiting microglia M1 polarization and targeting MAPK/NF-κB pathway / 陆军军医大学学报

Objective To explore the impact of the sialic acid binding lectin-E(Siglec-E)on the inhibitory properties of parthenolide(PTL)against lipopolysaccharide(LPS)-induced M1 polarization of microglia(BV2).Methods ①Single cell sequencing data of Siglece related mouse brain tissue was obtained from Gene Expression Omnibus(GEO)database and divided into the WT group(n=3)and the Siglece-/-group(n=4).The microglia cells were screened,and the enrichment analysis was performed to analyze related differential genes and pathways.BV2 cells were constructed by the shRNA interference technique and were divided into NC-shRNA and Siglece-shRNA to detect the expression level of Siglec-E(Siglece).② NC-shRNA and Siglece-shRNA cells were respectively divided into the Control group,LPS group,PTL group and PTL+LPS group(n=3).The mRNA levels of markers of M1 polarization in microglia,iNOS,IL-1 β and IL-6,were detected by RT-qPCR.Siglecefl/fl and Cx3cr1cre mice were mated to obtain microglia-specific Siglece deletion(Siglecefl/fl×Cx3cr1cre)mice,and LPS-induced neuroinflammation model was established.③ Nine WT and Siglecefl/fl×Cx3cr1cre male mice were assigned to the Control group,LPS group and PTL+LPS group(n=3).RT-qPCR,immunofluorescence assay and Western blotting were used to verify the knock-out effect and polarization-related pathways,and to investigate the mechanism of Siglec-E affecting PTL inhibition of M1 polarization of microglia.Results Compared with the NC-shRNA group,the expression of Siglec-E in the Siglece-shRNA group was significantly decreased(P<0.01),indicating that the Siglec-E knock-down cell model was successfully established.With the stimulation of LPS,mRNA levels ofiNOS,IL-1 β and IL-6 were significantly up-regulated compared with the Control group both in shRNA cells and Siglece-shRNA cells(P<0.01).With the influence of PTL and LPS,the markers of M1 polarization in NC-shRNA cells mentioned before were significantly decreased(P<0.05),while for Siglice-shRNA cells,there were no significant changes in the markers of M1 polarization.PTL inhibited the phosphorylation of JNK and IκB protein(P<0.01)and the nuclear translocation of NF-κB in BV2 cells,down-regulated Siglec-E,and weakened the inhibitory effect.Compared with mice in the WT group,the expression of Siglec-E in microglia of Siglecefl/fl×Cx3cr1cre mice was decreased significantly(P<0.01),and the inhibitory effect of PTL on the phosphorylation of NF-κB in microglia of Siglecefl/fl×Cx3cr1cre mice was also decreased.Conclusion The absence of Siglec-E in microglia attenuates the inhibition of M1 polarization by the MAPK/NF-κB pathway targeted by PTL.

Sialic acid-mediated photochemotherapy enhances infiltration of CD8+ T cells from tumor-draining lymph nodes into tumors of immunosenescent mice

Immunoscenescence plays a key role in the initiation and development of tumors. Furthermore, immunoscenescence also impacts drug delivery and cancer therapeutic efficacy. To reduce the impact of immunosenescence on anti-tumor therapy, this experimental plan aimed to use neutrophils with tumor tropism properties to deliver sialic acid (SA)-modified liposomes into the tumor, kill tumor cells via SA-mediated photochemotherapy, enhance infiltration of neutrophils into the tumor, induce immunogenic death of tumor cells with chemotherapy, enhance infiltration of CD8+ T cells into the tumor-draining lymph nodes and tumors of immunosenescent mice, and achieve SA-mediated photochemotherapy. We found that CD8+ T cell and neutrophil levels in 16-month-old mice were significantly lower than those in 2- and 8-month-old mice; 16-month-old mice exhibited immunosenescence. The anti-tumor efficacy of SA-mediated non-photochemotherapy declined in 16-month-old mice, and tumors recurred after scabbing. SA-mediated photochemotherapy enhanced tumor infiltration by CD8+ T cells and neutrophils, induced crusting and regression of tumors in 8-month-old mice, inhibited metastasis and recurrence of tumors and eliminated the immunosenescence-induced decline in antitumor therapeutic efficacy in 16-month-old mice via the light-heat-chemical-immunity conversion.

Free sialic acid storage disorders with fetal hydrops in a neonate / 中国当代儿科杂志

A boy, aged 3 hours, was admitted due to a prenatal diagnosis of fetal hydrops at 3 hours after resuscitation for birth asphyxia. Prenatal examination at 5 months of gestation showed massive ascites in the fetus, and after birth, the boy had the manifestations of systemic hydroderma, massive ascites, coarse face, and hepatomegaly. Genetic testing revealed heterozygous mutations in the SLC17A5 gene, and there was a significant increase in urinary free sialic acid. Placental pathology showed extensive vacuolization in villous stromal cells, Hofbauer cells, cytotrophoblast cells, and syncytiotrophoblast cells in human placental chorionic villi. The boy was finally diagnosed with free sialic acid storage disorders (FSASDs). This is the first case of FSASDs with the initial symptom of fetal hydrops reported in China. The possibility of FSASDs should be considered for cases with non-immune hydrops fetalis, and examinations such as placental pathology and urinary free sialic acid may help with early diagnosis and clinical decision making.

Co-load of silybin and doxorubicin by MoS2 nanosheets for synergetic chemotherapy and photothermal therapy of lung cancer / 药学学报

The active ingredient of traditional Chinese medicine, silybin (SBN), can inhibit the proliferation of cancer cells and enhance the anticancer effect of doxorubicin (DOX). However, due to non-targeting and short half-life of SBN and DOX, as well as different administration routes and pharmacokinetic processes, this combination drug cannot act on the tumor in the set order, seriously eliminating the synergistic effect between them and limiting the effect in vivo. Therefore, we intended to construct a nano-delivery system based on molybdenum disulfide (MoS2), modified by polyethylene glycol (PEG) and sialic acid (SA), and co-loaded with SBN and DOX. The system induced the release of combined drugs under the dual-stimulation of pH and near infra-red (NIR), increased the free concentration of intracellular drugs, so as to achieve the synergistic effect between them. The animal welfare and experimental procedures were in accordance with the regulations of the Animal Ethics Committee of Fujian University of Traditional Chinese Medicine. MoS2-PEG-SA-SBN/DOX circulated in vivo, and effectively accumulated at tumor sites through enhanced permeability and retention effect (EPR) and SA-mediated active targeting. Under near infrared light irradiation, MoS2-PEG-SA-SBN/DOX realized the combination of synergistic chemotherapy and photothermal therapy for tumor, thus achieving excellent anti-tumor effect in vivo. This study can provide a new idea and strategy for the clinical treatment of lung cancer. Taken together, MoS2-PEG-SA-SBN/DOX can offer a new idea and strategy for the clinical treatment of lung cancer.

The cases of twins with sialidosis type 1 / 中华神经科杂志

Sialidosis is a rare lysosomal storage disease caused by NEU1 gene mutation at 6p21.33. It is characterized by myoclonic, ataxia, epilepsy, and decreased vision. A pair of twins with sialidosis type 1 are reported to enrich clinicians ′ understanding of the disease, so as to improve the diagnosis and treatment. The proband was a 16-year-old male. The main symptom was intermittent limb involuntary trembling for 2 years, with paroxysmal loss of consciousness. Fundus examination showed cherry-red spots. His twin brother had similar symptoms, but the overall performance was mild. Whole exome sequencing results showed that both patients carried compound heterozygous mutations of c.239C>T (p.P80L) and c.803A>G (p.Y268C) in NEU1 gene, which were from their normal phenotype mother and father.

The Role of Siglecs in Immune Signaling Regulation and Immune Pathology / 中国生物化学与分子生物学报

Sialic acid-binding immunoglobulin-like lectins (Siglecs) are members of the immunoglobulin superfamily expressed in most white blood cells of the immune system. The Siglec is a kind of transmembrane proteins that can recognize sialic acid-containing ligands, and it is the most important subgroup of type I lectin. All Siglecs contain at least three domains, including the V-set Ig domain, C2-set Ig domain and transmembrane domain. Some Siglecs contain immune receptor tyrosine-based inhibitory motifs (ITIMs), which are used to transmit inhibitory signals and play an inhibitory role. There are also some Siglecs that do not contain intracellular domains, and they can transmit activation signals through basic amino acids in transmembrane domains to play an activation role. Siglecs not only participate in the regulation of activation, proliferation and apoptosis of immune cells, but also help the immune system to distinguish itself from non-itself by recognizing glycan ligands containing sialic acid. In recent years, studies have shown that Siglecs, as an immune checkpoint, play an important role in the immune regulation in human diseases such as cancer, asthma, allergy, Alzheimer’s disease and autoimmune diseases, and it has received extensive attention. Enhancing or blocking the interaction of sialic acid with Siglecs is an effective strategy for the treatment of cancer, infection, and other diseases. In this review, we describe the classification of Siglecs, their expression in different immune cells and their role in the regulation of immune cell signaling. Emphasis was placed on the role of Siglecs in disease and methods of targeting Siglecs for the treatment of human diseases.

Sialic acid as the potential link between lipid metabolism and inflammation in the pathogenesis of atherosclerosis

In the modern world, cardiovascular diseases have a special place among the most common causes of death. Naturally, this widespread problem cannot escape the attention of scientists and researchers. One of the main conditions preceding the development of fatal cardiovascular diseases is atherosclerosis. Despite extensive research into its pathogenesis and possible prevention and treatment strategies, many gaps remain in our understanding of this disease. For example, the concept of multiple low-density lipoprotein modifications was recently stated, in which desialylation is of special importance. Apart from this, sialic acids are known to be important contributors to processes such as endothelial dysfunction and inflammation, which in turn are major components of atherogenesis. In this review, we have collected information on sialic acid metabolism, analyzed various aspects of its implication in atherosclerosis at different stages, and provided an overview of the role of particular groups of enzymes responsible for sialic acid metabolism in the context of atherosclerosis.

Siglec-15 Promotes the Proliferation and Migration of Colorectal Cancer Cells and Inhibits the Infiltration of CD4 / 中国生物化学与分子生物学报

Sialic acid binding immunoglobulin type lectin-15 (Siglec-15), one of the Siglec gene family members, is a new immunosuppressive molecule. Siglec-15 is highly expressed in a variety of human tumor cells and tumor-associated macrophages, but the biological function of Siglec-15 in colorectal cancer (CRC) and its effect on the tumor immune microenvironment remains poorly understood. This study aimed to investigate the effects of aberrant expression of Siglec-15 on the biological behaviors of CRC cells and the infiltration of CD4

Venenos de serpentes do gênero Bothrops: impacto da glicosilação na complexidade dos proteomas e função de toxinas / Bothrops snake venoms: impact of glycosylation on the complexity of proteomes and toxins function

A variabilidade estrutural é uma característica das proteínas de venenos de serpentes, e a glicosilação é uma das principais modificações pós-traducionais que contribui para a diversificação de seus proteomas. Recentes estudos de nosso grupo demonstraram que venenos do gênero Bothrops são marcadamente definidos pelo seu conteúdo de glicoproteínas, e que a maioria das estruturas de N-glicanos dos tipos híbrido e complexo identificados em oito venenos deste gênero contêm unidades de ácido siálico. Em paralelo, em glicoproteínas do veneno de B. cotiara foi identificada a presença de uma estrutura de N-acetilglicosamina bissecada. Assim, com o objetivo de investigar a variação do conteúdo de glicoproteínas, assim como os mecanismos envolvidos na geração dos diferentes venenos de Bothrops, neste estudo foram analisados comparativamente os glicoproteomas de nove venenos do gênero Bothrops (B. atrox, B. cotiara, B. erythromelas, B. fonsecai, B. insularis, B. jararaca, B. jararacussu, B. moojeni e B. neuwiedi). As abordagens glicoproteômicas envolveram cromatografia de afinidade e ensaio de pull-down utilizando, respectivamente, as lectinas SNA (aglutinina de Sambucus nigra) e MAL I (lectina de Maackia amurensis), que mostram afinidade por unidades de ácido siálico nas posições, respectivamente, α2,6 e α2,3; e cromatografia de afinidade com a lectina PHA-E (eritroaglutinina de Phaseolus vulgaris), que reconhece N-acetilglicosamina bissecada. Ainda, eletroforese de proteínas, blot de lectina, e identificação de proteínas por espectrometria de massas foram empregadas para caracterizar os glicoproteomas. As lectinas geraram frações dos venenos enriquecidas de diferentes componentes, onde as principais classes de glicoproteínas identificadas foram metaloprotease, serinoprotease, e L-amino ácido oxidase, além de outras enzimas pouco abundantes nos venenos. Os diferentes conteúdos de proteínas reconhecidas por essas lectinas, com especificidades distintas, ressaltaram novos aspectos da variabilidade dos subproteomas de glicoproteínas desses venenos, dependendo da espécie. Ainda, considerando que metaloproteases e serinoproteases são componentes abundantes nesses venenos e fundamentais no quadro de envenenamento botrópico, e que estas enzimas contêm diversos sítios de glicosilação, o papel das unidades de ácido siálico na atividade proteolítica das mesmas foi avaliado. Assim, a remoção enzimática de ácido siálico (i) alterou o padrão de gelatinólise em zimografia da maioria dos venenos, (ii) diminuiu a atividade proteolítica de alguns venenos sobre o fibrinogênio e a atividade coagulante do plasma humano de todos os venenos, e (iii) alterou o perfil de hidrólise de proteínas plasmáticas pelo veneno de B. jararaca, indicando que este carboidrato pode desempenhar um papel na interação das proteases com seus substratos proteicos. Em contraste, o perfil da atividade amidolítica dos venenos não se alterou após a remoção de ácido siálico e incubação com o substrato Bz-Arg-pNA, indicando que ácido siálico não é essencial em N-glicanos de serinoproteases atuando sobre substratos não proteicos. Em conjunto, esses resultados expandem o conhecimento sobre a variabilidade de proteomas de venenos do gênero Bothrops e apontam a importância das cadeias de carboidratos contendo ácido siálico nas atividades enzimáticas das proteases desses venenos

Structural variability is a feature of snake venom proteins, and glycosylation is one of the main post-translational modifications that contributes to the diversification of venom proteomes. Recent studies by our group have shown that Bothrops venoms are markedly defined by their glycoprotein content, and that most hybrid and complex N-glycan structures identified in eight venoms of this genus contain sialic acid units. In parallel, the presence of a bisected N-acetylglucosamine structure was identified in B. cotiara venom glycoproteins. Thus, with the aim of investigating the variation in the content of glycoproteins, as well as the mechanisms involved in the generation of different Bothrops venoms, in this study the glycoproteomes of nine Bothrops venoms (B. atrox, B. cotiara, B. erythromelas, B. fonsecai, B. insularis, B. jararaca, B. jararacussu, B. moojeni e B. neuwiedi) were comparatively analyzed. The glycoproteomic approaches involved affinity chromatography and pulldown using, respectively, the lectins SNA (Sambucus nigra agglutinin) and MAL I (Maackia amurensis lectin), which show affinity for sialic acid units at positions, respectively, α2,6 and α2,3, and affinity chromatography with PHA-E (Phaseolus vulgaris erythroagglutinin), which recognizes bisected N-acetylglucosamine. In addition, protein electrophoresis, lectin blot, and protein identification by mass spectrometry were employed for glycoproteome characterization. The lectins generated venom fractions enriched with different components, where the main classes of glycoproteins identified were metalloprotease, serine protease, and L-amino acid oxidase, in addition to other low abundant enzymes. The different contents of proteins recognized by these lectins of distinct specificities highlighted new aspects of the variability of the glycoprotein subproteomes of these venoms, depending on the species. Furthermore, considering that metalloproteases and serine proteases are abundant components of these venoms and essential in Bothrops envenomation, and that these enzymes contain several glycosylation sites, the role of sialic acid units in their proteolytic activities was evaluated. Thus, enzymatic removal of sialic acid (i) altered the pattern of gelatinolysis in zymography of most venoms, (ii) decreased the proteolytic activity of some venoms on fibrinogen and the clotting activity of human plasma of all venoms, and (iii) altered the hydrolysis profile of plasma proteins by B. jararaca venom, indicating that this carbohydrate may play a role in the interaction of proteases with their protein substrates. In contrast, the profile of amidolytic activity of the venoms did not change after removal of sialic acid and incubation with the substrate Bz-Arg-pNA, indicating that sialic acid is not essential in N-glycans of serine proteases acting on small substrates. Together, these results expand the knowledge about the variability of proteomes of Bothrops venoms and point to the importance of carbohydrate chains containing sialic acid in the enzymatic activities of venom proteases

Evaluations of antioxidant enzyme activities, total sialic acid and trace element levels in coronary artery bypass grafting patients

Abstract Introduction: In this study, patients before and after cardiac surgery with cardiopulmonary bypass (CPB) and control subjects were evaluated for erythrocyte glutathione peroxidase, catalase and superoxide dismutase enzyme activities, in addition to glutathione, malondialdehyde, serum total sialic acid, lipid-bound sialic acid, total antioxidant status, trace elements and mineral levels. The correlation of these variables with coronary artery disease (CAD) was also assessed. Methods: A total of 30 CAD patients and 30 control subjects were included in the study. CAD patients were divided into three groups: before surgery (BS), first day after surgery (1st day AS) and seventh day after surgery (7th day AS). Results: Malondialdehyde (MDA) and total sialic acid (TSA) levels were significantly higher in CAD (BS) than in the control group (P<0.05, P<0.05). In addition, GSH and TAS levels were significantly lower in the 1st day AS group than in the control group (P<0.001, P<0.01). Moreover, Co, Cu, Mg, Se, V and Zn levels were significantly lower in CAD (BS) group than in the control group (P<0.01, P<0.01, P<0.01, P<0.01, P<0.05, P<0.001). Conclusions: It was concluded that the levels of LDL-C, total cholesterol, triglycerides and CRP significantly associated with parameters, as well as Cu, Ca and SOD activity, should be measured together to monitor CAD. It is also considered that measuring TSA and MDA might be an appropriate choice for biomarkers of CAD.

Effect of fangchinoline on oxidant status in male albino rats with streptozotocin-induced diabetes

BACKGROUND Diabetes is a metabolic disorder caused by defects in insulin production and activity. During disease progression, changes in lipid peroxidation cause structural modifications via production of free radicals. Fangchinoline is a well-known alkaloid present in Stephaniae tetrandrine S. Moore, which has demonstrated antioxidant, anticancer, and anti-inflammatory activities. RESULTS The present study analyzed the anti-diabetic and antioxidant effects of fangchinoline in male rats with streptozotocin-induced diabetes. Rats were divided into the following groups: normal control, diabetic, diabetic + fangchinoline 100 mg/kg, diabetic + fangchinoline 200 mg/kg and diabetic + glibencla mide 600 mg/kg. The treatment was administered orally for 45 consecutive days. Lipid peroxidation was substantially increased by >50% in the serum, as well as the liver, kidney, and heart tissues of diabetic rats. However, fangchinoline supplementation significantly reduced lipid peroxidation to near normal levels. Reactive oxygen species levels were substantially increased by >500% in the serum, as well as the liver, kidney, and heart tissues of diabetic rats. Fangchinoline supplementation reduced reactive oxygen species to near normal levels. Fangchinoline supplementation significantly improved superoxide dismutase, glutathione peroxidase, catalase, and reduced glutathione levels in diabetic rats. Total hexoses, sialic acid, hexosamines, and fucose were increased in diabetic rats, whereas fangchinoline supplementation significantly reduced these total hexoses, sialic acid, hexosamines, and fucose to near normal levels CONCLUSIONS Supplementation with fangchinoline led to significant attenuation of the levels of lipid peroxidation, ROS, and glycoprotein components such as total hexoses, hexosamines, sialic acid, and fucose, while improving antioxidant marker levels

Uso do gérmen de milho desengordurado, com e sem adição de um complexo enzimático, em dietas para cães / [Use of defatted corngerm with and without the addition of an enzyme complex in dog diets]

Objetivou-se avaliar o coeficiente de digestibilidade aparente (CDA) dos nutrientes, a palatabilidade das dietas e as características fecais de cães alimentados com uma dieta controle e uma dieta contendo 20% de gérmen desengordurado (GD), com e sem adição de complexo enzimático (amilase, xilanase, betaglucanase e mananase). Para o experimento de digestibidade e das características fecais, foram utilizados 12 cães adultos, distribuídos em delineamento em blocos ao acaso, em esquema fatorial 2 x 2 (dieta x enzima). O segundo experimento avaliou a palatabilidade, por meio da primeira escolha e da razão de ingestão (RI) da dieta DC vs. 20% de GD, utilizando-se 16 cães. O teste de palatabilidade contou com três dias consecutivos, totalizando 48 repetições. A dieta com inclusão de 20% de GD teve os menores valores de CDA da MS, da EB e da EM (P<0,05). A inclusão do complexo enzimático melhorou o CDA da MS, da EB e da EM (P<0,05). Não foram observadas diferenças nas características fecais (P>0,05). Em relação à palatabilidade, os cães preferiram a dieta 20% de GD, tanto na primeira escolha como na RI (P<0,05). A inclusão de enzimas às dietas melhora a digestibilidade dos nutrientes e da EM, sendo um aditivo com potencial uso na alimentação de cães.(AU)

The objective was to evaluate the apparent digestibility coefficient (ADC) of nutrients, diet palatability and fecal characteristics of dogs fed diets containing degreased germ (DG), and a control diet (DC) - both with and without the addition of enzyme complex (amylase, xylanase, betaglucanase and mananase). For the digestibility and fecal characteristics experiment 12 adult dogs were used, distributed in a randomized block design, in a 2 x 2 factorial scheme (diet x enzyme). The second experiment evaluated palatability using the first choice and ingestion ratio (IR) of DC diet vs. 20%gD, using 16 dogs. The palatability test had three consecutive days, totaling 48 repetitions. The diet with inclusion of 20% DG had the lowest ADC values of DM, GE and ME (P <0.05). Inclusion of the enzyme complex improved ADC of DM, GE and ME (P <0.05). No differences in fecal characteristics were observed (P >0.05). Regarding palatability, dogs preferred the 20% DG diet in both first choice and IR (P <0.05). Inclusion of enzymes in diets improves nutrient digestibility and ME, being an additive with potential use in dog food.(AU)

Relevance of serum and salivary sialic acid in oral cancer diagnostics

To review the relevance of sialic acid as a tumour marker in oral cancer. Tumour marker are useful in the screening for early malignancy. Sialic acids are important in determining the surface properties of cells and has been implicated in cellular invasiveness, adhesiveness, and immunogenicity. Sialic acids are commonly found at the outermost end of glycan chains of all cell types. Increase in the levels of sialic acid in oral cancer indicates its importance as a tumour marker.Both serum and salivary sialic acid levels can be used as a screening tool and a diagnostic aid for oral cancer. Salivary sialic acid can be used as a non-invasive, cost effective and reliable diagnostic methods for screening and monitoring of oral cancer. In patients with oral cancer, glycoprotein metabolism is altered. Increase in the levels of sialic acid in oral cancer indicate its importance as a tumour marker. Changes in the serum is reflected in saliva. Salivary sialic acid can be used as non-invasive, cost effective and reliable diagnostic methods for screening and monitoring of oral cancer. Early the diagnosis, better the prognosis

Usefulness of salivary sialic acid as a tumor marker in tobacco chewers with oral cancer

Aim:This study aims to assess the usefulness of salivary sialic acid (SA) as a tumor marker in the detection of oral squamous cell carcinoma (OSCC) among tobacco chewers. Materials and Methods:After the approval of study protocol by the Institutional Ethics Committee and informed voluntary consent, salivary samples were collected from 96 participants in each group of tobacco chewers with OSCC, tobacco chewers without precancerous or cancerous lesion, and healthy controls. Salivary protein-bound SA (PBSA) and salivary-free SA (FSA) were measured by Yao et al.'s method of acid ninhydrin reaction, and the data were subjected to appropriate statistical analysis. Results: The salivary PBSA and FSA levels in the Groups 1, 2, and 3 participants were 31.17 ± 7.6 mg/dL and 63.45 ± 9.8 mg/dL, 25.45 ± 16.61 mg/dL and 33.18 ± 11.38 mg/dL, and 22.73 ± 3.01 mg/dL and 21.62 ± 8.86 mg/dL, respectively. Salivary FSA levels were significantly increased among the tobacco chewers with OSCC patients (Group 1) and tobacco chewers with no premalignant lesions of the oral cavity (Group 2) compared to the healthy controls (Group 3) with P < 0.05 being statistically significant. Salivary FSA levels were significantly increased in Group 1 as compared with Group 2. The salivary PBSA was high among Group 1 as compared to the control Group 3; there was however no significant difference in the levels of salivary PBSA between Group 1 and Group 2. There was no significant difference in the PBSA levels between OSCC patients of Group 1 and the tobacco chewers without precancerous or cancerous lesion in the oral cavity of Group 2. Conclusion: Salivary PBSA and FSA are significantly raised in both tobacco chewers with OSCC and in tobacco chewers with no precancerous or cancerous lesions in the oral cavity. SA should therefore be used cautiously while considering it as a marker for the early detection of oral cancer. Tobacco can be a crucial confounding factor when SA is used as a biomarker in OSCC since their levels are elevated to some extent even in tobacco chewers without any clinically obvious precancerous or cancerous lesions in the oral cavity

Analysis of WCID-cIEF Spectrogram of Recombinant Interleukin-15 Fusion Protein / 中国药学杂志

OBJECTIVE: To analyze the glycosylated chains of recombinant interleukin-15 fusion protein using capillary isoelectric focusing-whole column imaging detection (WCID-cIEF) spectrograms. METHODS: Using established corresponding mathematical models and the least square method, the WCID-cIEF spectrograms of whole protein, de-salicylic-acid protein and de-N-glycosylation-chain protein were analyzed. Among the mathematical models, the interval-1-peak model was selected. And according to the model, the relationship between isoform peak-areas and isoelectric points was listed. RESULTS: The rationality of the interval-1-peak model was confirmed and a series of basic data was obtained according to the model as follows:the apparent m value of the protein was 25.53 reference(R), the apparent n value of the protein was 28.83R, the apparent m value of sialic acid was 0.86 (0.855) R, the apparent n value was 0.12 (0.119) R, the apparent n value of N-acetylglucosamine (undifferentiated from N-acetylgalactosamine) was 0.06(0.061) R, and the apparent m value of formed carboxyl after N-chain removal was 0.19 (0.186) R. Some information of protein sugar composition was also obtained: the sialylation degree was about 1.83 mol•mol-1, the percentage of prototype protein was about 8.3%, the percentage of single N-glycosated modification protein was about 19.8%, the percentage of double N-glycosated modification protein was about 28.4%, the percentage of triple N-glycosated modification protein was about 23.7%, and the percentage of O-glycosated modification (with sialic acid) protein was about 19.8%. The main sugar types should be G0 (F), G1 (F), G2 (F), G1A1 (F), and G2A1 (F). CONCLUSION: The structure of sugar chain is complex, but it also has some repeatability and regularity. We hope that through this study, the glycoprotein sugar chain can be quickly outlined, the understanding of glycoprotein and the study of protein interaction can be improved.

Preparation of sialic acid-modified chlorogenic acid liposomes and its anti-tumor activity in vitro / 中草药

Objective: Sialic acid (SA)-modified chlorogenic acid (CA) liposomes (CA-SAL) was prepared by response surface design to investigate its in vitro cytotoxicity and uptake. Methods: CA-SAL was prepared by a modified reverse-phase ethanol injection method. Sephadex G50 column was used to separate the CA-loaded liposomes and the free CA. The drug concentration was determined by HPLC method and the encapsulation efficiency was calculated. With encapsulation efficiency and drug loading as indicators, Box-Behnken response surface design experiments were used to optimize the prescription process of CA-SAL. The MTT method was used to evaluate the cytotoxicity of CA-SAL on human lung cancer cells A549. Inverted fluorescence microscope was used to investigate the uptake of CA-SAL by A549 cells. Results: The optimized preparation conditions: hydrogenated soybean lecithin-CA ratio at 15:1, hydration temperature 60 ℃, ultrasonic power 400 W. The average particle size of CA-SAL was (90.13 ± 0.51) nm, the polydispersity index (PDI) was 0.16 ± 0.01, the zeta potential was (-25.3 ± 0.5) mV, the encapsulation efficiency was 57.8%, RSD was 0.1%. MTT results showed that the inhibitory effect of CA-SAL on A549 cells was significantly greater than CA-CL. Greater cellular uptake of CA-SAL was observed compared with CA-CL. Conclusion: CA-SAL prepared by response surface optimization has a uniform particle size and good stability. SA-modified CA-loaded liposomes could enhance cellular uptake and cytotoxicity of human lung cancer cell A549 in vitro.

Acrylamide reduces plasma antioxidant vitamin levels in rats due to increased oxidative damage / Acrilamida reduz os níveis de vitamina antioxidante no plasma em ratos devido ao aumento do dano oxidativo

ABSTRACT Objective Acrylamide is a potentially neurotoxic and carcinogenic chemical and naturally creates during the heating process of carbohydrate-rich foods, such as potato chips and breakfast cereals. Acrylamide might be ingested by people via consuming food that contains it. Therefore, we investigated the effect of acrylamidegiven orally to male and female rats on plasma retinoic acid and α-tocopherol and serum sialic acid and malondialdehyde levels. Method A total of 50 Wistar rats were used (25 female and 25 male, three-four weeks old). The rats of each sex were given 2 and 5mg/kg/day acrylamide via drinking water for 90 days. At the end of the treatment, the animals were euthanized by cervical dislocation. Blood specimens were collected through cardiac puncture, and serum and plasma samples were analysed using the high-performance liquid chromatography technique with a Ultraviolet detector. Results The analysis of the plasma and serum samples revealed that serum sialic acid and malondialdehyde levels in both sexes given 5mg/kg/day acrylamide were significantly increased, and the serum sialic acid levels were higher in female rats given 2mg/kg/day acrylamide. The plasma retinoic acid and α-tocopherol levels significantly decreased in both sexes given only the highest dose. Conclusion The results show that acrylamide causes an increase in oxidative stress and leads to a decrease in the levels of retinoic acid and α-tocopherol which play a role in the defense mechanism against this stress.

RESUMO Objetivo A acrilamida é um químico potencialmente neurotóxico e carcinogênico, sendo naturalmente criada durante o processo de aquecimento de alimentos ricos em carboidratos, como batatas fritas e cereais matinais. Dado que o composto pode ser ingerido através do consumo de alimentos, o presente trabalho teve por objetivo investigar o seu efeito, quando administrado oralmente a ratos, medindo-se os níveis plasmáticos de ácido retinoico e α-tocoferol, bem como os níveis séricos de ácido siálico e malondialdeído Métodos Foram utilizados cinquenta ratos Wistar, sendo metade de cada sexo, com idade entre três e quatro semanas. Os animais foram divididos em dois grupos, os quais receberam diferentes doses diárias de acrilamida, via água potável, durante noventa dias: o primeiro ingeriu 2mg/kg/dia; e o segundo, 5mg/kg/dia. Ao final do tratamento, os animais foram eutanasiados por meio de luxação cervical. Amostras de sangue foram coletadas através de punção cardíaca, assim como amostras de soro e plasma foram medidas usando-se a técnica de cromatografia líquida de alta performance com detector de Ultravioleta. Resultados A análise das amostras de plasma e soro revelou que os níveis de ácido siálico e malondialdeído, em ratos de ambos os sexos tratados com acrilamida de 5mg/kg/dia, foram significativamente aumentados, ao passo que os níveis séricos de ácido siálico foram maiores em ratas tratadas com 2mg/kg/dia de acrilamida. Já os níveis plasmáticos de ácido retinoico e α-tocoferol diminuíram significativamente em ratos de ambos os sexos, quando tratados com a dose mais elevada.Concl Conclusão Os resultados mostram que a acrilamida causa um aumento no estresse oxidativo e leva a uma diminuição nos níveis de ácido retinoico e α-tocoferol, que desempenham um papel no mecanismo de defesa contra esse estresse.

New Rabbit Erythrocyte Specific Mycelial Lectins from Fusarium sp. with Complex Saccharide Specificity

Eight Fusarium sp. namely, F. acutatum, F. globosum, F. graminearum, F. lactis, F. nivale, F. proliferatum, F.pseudoanthophilum and F. robustum were screened for the presence of lectins by hemagglutination activityusing human ABO, porcine, ovine, goat and rabbit erythrocytes. Mycelial extracts of all the fungal culturesexcept F. graminearum displayed unique lectin activity with only rabbit erythrocytes. Enzymatic treatment ofrabbit erythrocytes with neuraminidase has significantly enhanced the titre of all the lectin-positive extractsof fungal cultures. In contrast, most of the lectins showed a decline in lectin activity with protease treatedrabbit erythrocytes. Saccharide specificity studies have shown that majority of the lectins are inhibitory towardsO-acetyl sialic acids. None of the lectins from Fusarium sp. were inhibited by dextran, meso-inositol, andN-acetyl-D-glucosamine. Most of the fungal cultures displayed highest hemagglutination activity during the10th day of growth in broth cultures. The unique saccharide specificity of Fusarium sp. lectins can be used forelucidating their clinical role in glycobiology research.

The significance of Hp antigen combined with serum total sialic acid and gastric mucosal Survivin detection in the diagnosis of gastric adenocarcinoma / 国际检验医学杂志

Objective To explore the significance of Hp antigen combined with serum total sialic acid and gastric mucosal Survivin detection in the diagnosis of gastric adenocarcinoma.Methods Selected a hospital 90cases of patients with gastric adenocarcinoma (malignant group) and 90cases of the digestive system benign disease (benign disease group) and hospital check-up of 60cases of healthy people (healthy control group) as the research object.Comparing the three groups of patients with Hp antigen, serum total sialic acid positive detection rate and the positive rate of Survivin in the gastric mucosa, and to assess individual detection of Hp antigen, serum total sialic acid and the two combined diagnostic sensitivity and specificity of gastric adenocarcinomas.Results Hp antigen, serum total sialic acid positive test results showed that the malignant tumor group were significantly higher than that of benign disease group and healthy control group, benign disease group obviously higher than that of healthy control group, difference between three groups of participants were statistically significant (P<0.05) .The sensitivity of Hp antigen and serum total sialic acid combined in the diagnosis of digestive system was 80.1％, which was significantly higher than that of single test, but the specificity was reduced.Linear correlation analysis showed that there was a positive correlation between Hp antigen and serum total sialic acid and Survivin detection in gastric mucosa, and the correlation was high (r=0.94, P<0.05) .Conclusion The combined detection of Hp antigen and total serum sialic acid is positively correlated with the results of Survivin detection in gastric mucosa, and has a high correlation.It can replace Survivin detection in gastric mucosa as a sensitive index for the diagnosis of gastric adenocarcinoma.

Levels of Salivary Sialic Acid in Children with Autism Spectrum Disorder; Could It Be Related to Stereotypes and Hyperactivity?

OBJECTIVE: Sialic acid (Sia) is an essential nutrient for brain development, learning, memory and cognition and plays a role in neurodevelopment of infants. The aim of this study was to determine whether Sia levels are significantly associated with the autism spectrum disorder (ASD). METHODS: Forty-six ASD children and 30 typically developing children aged 3 to 10 years were included in the study. Behavioral symptoms in ASD children was assessed by the Autism Behavior Checklist (AuBC), the Childhood Autism Rating Scale, and the Aberrant Behavior Checklist (ABC). After the collection of saliva samples, the supernatant was separated. All the samples kept at −80°C until Sia analysis was done. RESULTS: Sia level was found to be significantly lower in the ASD group when compared to healthy controls (p = 0.013). There was no correlation between severity of ASD and salivary Sia levels. We found a negative correlation between AuBC scores and Sia levels and a negative correlation in both ABC Stereotypic Behavior and Hyperactivity/Noncompliance subscales with Sia levels in ASD group. CONCLUSION: The obtained data indicate that Sia levels could have an effect on autism-like behaviors, particularly on stereotypes and hyperactivity.