Beneficial or detrimental: Recruiting more types of benign cases for cancer diagnosis based on salivary glycopatterns.

With the advantages of convenient, painless and non-invasive collection, saliva holds great promise as a valuable biomarker source for cancer detection, pathological assessment and therapeutic monitoring. Salivary glycopatterns have shown significant potential for cancer screening in recent years. However, the understanding of benign lesions at non-cancerous sites in cancer diagnosis has been overlooked. Clarifying the influence of benign lesions on salivary glycopatterns and cancer screening is crucial for advancing the development of salivary glycopattern-based diagnostics. In this study, 2885 samples were analyzed using lectin microarrays to identify variations in salivary glycopatterns according to the number, location, and type of lesions. By utilizing our previously published data of tumor-associated salivary glycopatterns, the performance of machine learning algorithm for cancer screening was investigated to evaluate the effect of adding benign disease cases to the control group. The results demonstrated that both the location and number of lesions had discernible effects on salivary glycopatterns. And it was also revealed that incorporating a broad range of benign diseases into the controls improved the classifier's performance in distinguishing cancer cases from controls. This finding holds guiding significance for enhancing salivary glycopattern-based cancer screening and facilitates their practical implementation in clinical settings.

A novel strategy for quantification of α2,3- and α2,6-linked sialic acids in sialylated glycoproteins.

Sialic acid, a monosaccharide containing nine carbon atoms, is widely distributed in eukaryotic cells. The bound sialic acids are mainly present at the glycan ends of glycoconjugates via α2-3 or α2-6 glycosidic bonds, and alterations in their expression levels and linkage types are associated with the progress of many diseases and tumors. The present study provides a new strategy for quantification of α2,3- and α2,6-linked sialic acids in sialylated glycoproteins. In fact, quantification of α2,3-linked sialic acids were based on the difference of the bound sialic acids in the sample before and after treatment with α2-3 neuraminidase, whereas the α2,6-linked sialic acids were equal to the bound sialic acids in the α2-3 neuraminidase-treated sample. Subsequently, α2,3/6-linked sialic acids in salivary glycoproteins from healthy volunteers and diabetic patients were quantified in accordance with this method. This work provides an accurate method for the quantification of α2,3- and α2,6-linked sialic acids in the sialoglycoproteins, which is more instructive for understanding the biological roles of α2,3/6-linked sialic acid in sialoglycoproteins.

Roles of bovine sialoglycoproteins for anti-skin aging and accelerating skin wound healing.

BACKGROUND: Fibroblasts are the most predominant cell subpopulation in the dermal layer of human skin, they play an important role in maintaining skin architecture and function. The senescence of fibroblasts is one of major causes of skin aging and chronic wound in the elderly, which is accompanied with a reduction of α2,6-sialylation on the cell surface. AIMS: In this study, we investigated the effects of the bovine sialoglycoproteins on normal human dermal fibroblasts (NHDF). RESULTS: The results showed that bovine sialoglycoproteins could promote the proliferation and migration of NHDF cells, and accelerate the contraction of fibroblast-populated collagen lattice (FPCL). The average doubling time of NHDF cells treated with bovine sialoglycoproteins (0.5 mg/mL) was 31.1 ± 1.0 h whereas that was 37.9 ± 2.7 h for the control (p ˂ 0.05). Moreover, the expression of basic fibroblast growth factor (FGF-2) was upregulated, while that of transforming growth factor-beta 1 (TGF-ß1) and human type I collagen (COL-I) were downregulated in treated NHDF cells. Furthermore, bovine sialoglycoproteins treatment significantly enhanced the α2,6-sialylation on the cell surfaces, which was consistent with the upregulation of α2,6-sialyltransferase I (ST6GAL1) expression. CONCLUSIONS: These results indicated that the bovine sialoglycoproteins might be developed as a reagent against skin aging in the cosmetic industry, or as a new candidate for accelerating skin wound healing and inhibiting scar formation.

Integrated glycomics strategy for galactosylated-N-glycans recognized by Bandeiraea Simplicifolia Lectin I in salivary proteins associated with lung cancer.

PURPOSE: Lung cancer (LC) is the leading cause of cancer-related deaths worldwide, mainly due to late diagnosis and poor prognosis. Saliva is an important source for discovering biomarkers and contains an abundance of biological information. The purpose of this study was to determine whether galactosylation levels of salivary proteins are associated with LC. EXPERIMENTAL DESIGN: First, we analyzed the alterations of the glycopatterns recognized by Bandeiraea Simplicifolia Lectin I (BS-I) in five groups (healthy volunteers [HV]: 28, benign pulmonary disease [BPD]: 27, lung adenocarcinoma [ADC]: 39, squamous cell carcinoma [SCC]: 28, small-cell lung cancer [SCLC]: 22) of 144 saliva samples using lectin microarrays. Pooled samples from each group were subsequently validated by the lectin blotting technique. Finally, the N-glycan profiles of their salivary glycoproteins isolated by the BS-I-magnetic particle conjugates from pooled samples for each group were analyzed by MALDI-TOF/TOF-MS. RESULTS: The results showed that the expression level of galactosylated glycans recognized by BS-I was significantly increased in patients with LC compared with BPD and HV. Receiver operating characteristic (ROC) analysis indicated that the levels of salivary glycopattern recognized by BS-I could discriminate lung disease (BPD, ADC, SCC, and SCLC) and HV with an AUC of 0.700 (95% CI: 0.589-0.812), and discriminate LC and BPD with an AUC of 0.860 (95% CI: 0.763-0.956). Also, the proportion of galactosylated N-glycans in ADC (38.4%), SCC (43.1%), and SCLC (39.5%) increased compared to HV (30.1%) and BPD (33.7%), and two galactosylated N-glycan peaks (m/z 1828.683, 2418.853) could be identified only in the LC groups (ADC, SCC, and SCLC). CONCLUSIONS AND CLINICAL RELEVANCE: These findings could provide crucial information on galactosylated N-linked glycans associated with LC and facilitate the study of LC biomarkers based on precise alterations of galactosylated N-glycans in saliva.

Machine learning reveals salivary glycopatterns as potential biomarkers for the diagnosis and prognosis of papillary thyroid cancer.

The diagnosis of thyroid cancer, especially papillary thyroid cancer (PTC), is increasing rapidly worldwide. In this study, we aimed to study the glycosylation of salivary proteins associated with PTC and assess the likelihood that salivary glycopatterns may be a potential biomarker of PTC diagnosis. Firstly, 22 benign thyroid nodule (BTN) samples, 27 PTC samples, and 30 healthy volunteers (HV) samples were collected to probe the difference of salivary glycopatterns associated with PTC using lectin microarrays. Then, five machine learning models including K-Nearest Neighbor (KNN), Multilayer Perceptron (MLP), Logistic Regression (LR), Random Forest (RF), and Support Vector Machine (SVM) were established to distinguish HV, BTN and PTC based on the changes of salivary glycopatterns. As a result, SVM had the best diagnostic effect with an accuracy rate of 92 % in testing set. Besides, lectin microarrays were used to explore the differences in salivary glycopatterns of 26 paired salivary samples of PTC patients before and after operation in order to probe into salivary glycopatterns as potential biomarkers for prognosis of PTC patients. The results showed that the levels of salivary glycopatterns recognized by 6 different lectins in patients after the operation almost convergenced with HVs. This study could help to screen and assess patients with PTC and their prognosis based on precise changes of salivary glycopatterns.

Heat stress response in Chinese cabbage (Brassica rapa L.) revealed by transcriptome and physiological analysis.

High temperatures have a serious impact on the quality and yield of cold-loving Chinese cabbage, which has evolved to have a unique set of stress mechanisms. To explore the relationship between these mechanisms and the heat-tolerance of Chinese cabbage, the physiological indicators of the heat-tolerant '268' line and heat-sensitive '334' line were measured. Under heat stress, the proline (Pro), soluble sugar (SS), and superoxide dismutase (SOD) indexes of the '268' line increased significantly. When additionally using transcriptome analysis, we found that the identified 3,360 DEGs were abundantly enriched in many metabolic pathways including 'plant hormone signal transduction', 'carbon metabolism', and 'glycolysis/gluconeogenesis'. Dynamic gene expression patterns showed that HKL1 in Cluster 15 may be a key factor in the regulation of sugar homeostasis. The interaction network screened four ABA-related genes in Cluster 15, suggesting that high temperatures lead to changes in hormonal signaling, especially an increase in ABA signaling. Compared with the '334' line, the expressions of Prx50, Prx52, Prx54, SOD1, and SOD2 in the '268' line were significantly upregulated, and these genes were actively involved in the reactive oxygen species (ROS) scavenging process. In summary, our results revealed the relationship between plant heat tolerance, physiology, and biochemistry and may also provide ideas for the future development of high-quality and heat-tolerant Chinese cabbage germplasm resources.

Genetic dissection of heterotic loci associated with plant weight by Graded pool-seq in heading Chinese cabbage (Brassica rapa).

MAIN CONCLUSION: Four heterotic QTL and a heterozygous segment for plant weight were identified by Graded Pool-Seq, QTL-seq and traditional genetic linkage analysis in heading Chinese cabbage. Heading Chinese cabbage (Brassica rapa L. spp. pekinensis) is a cross-pollinated leafy vegetable with significant heterosis. The use of heterosis is important for breeding high-yield Chinese cabbage hybrids. However, the formation and mechanism of heterosis have not been studied. We dissected the molecular mechanism of heterosis of yield-related traits in Chinese cabbage. An F1 hybrid with high-parent heterosis of yield-related traits was selected and self-pollinated to generate segregating F2 populations. QTL-seq, Graded Pool-seq (GPS), and traditional genetic linkage analysis were used to identify four heterotic quantitative trait loci (QTL) for plant weight: qPW1.1, qPW5.1, qPW7.1, and qPW8.1. Traditional genetic linkage analysis over two years showed that qPW8.1, located in marker A08_S45 (18,172,719) and A08_S85 (18,196,752), was mapped to a 23.5 kb genomic region. QTL qPW8.1 explained 8.6% and 23.6% of the phenotypic variation in plant weight and the total numbers of head leaves, respectively, and contained a heterozygous segment that might control the heterosis of plant weight. The qPW1.1 made an 11.7% phenotypic contribution to plant weight. The qPW7.1 was sensitive to environmental influence and explained 10.7% of the phenotypic variance. QTL qPW5.1 had a significant signal and was located in a genetic region near the centromere showing high heterozygosity. The "pseudo-overdominance" and "synergistic allelic" effects from parent line "XJD4" appear to play an important role in heterosis for plant weight in Chinese cabbage. These results provide a basis for an improved understanding of the molecular mechanism of yield-related traits and their heterosis.

Fingerprint construction through genotyping by sequencing for applied breeding in Brassica rapa.

This study evaluated the genotyping by sequencing (GBS) protocol for fingerprinting Brassica rapa, and the data derived were more reliable than the re-sequencing data of B. rapa. Of the 10 enzyme solutions used to analyze the numbers of genotypes and single-nucleotide polymorphisms (SNPs) in B. rapa, five solutions showed better results, namely, A (HaeIII, 450-500 bp), E (RsaI+HaeIII, 500-550 bp), F (RsaI+HaeIII, 500-600 bp), G (RsaI+HaeIII, 'All' fragment), and J (RsaI+EcoRV-HF®, 'All' fragment). The five enzyme solutions showed less than 40% similarity in different individuals from various samples, and 90% similarity between two individuals from one sample. The E enzyme solution was the most suitable for fingerprinting B. rapa, revealing well-distributed SNPs in the whole genome. Of the 82 highly inbred lines and 18 F1 lines of B. rapa sequenced by GBS in the E enzyme solution, known parents of 10 F1 lines were verified, and male parents were discovered for 8 F1 lines that had only known female parents. This study provides a valuable method for screening parents for F1 lines in B. rapa for the efficient evaluation of GBS with varied library construction strategies.

Role of sialylated glycans on bovine lactoferrin against influenza virus.

Influenza is a worldwide plague caused by the influenza virus (IAV) infection, which is initiated by specific recognition with sialic acids on host cell surface. Bovine lactoferrin (bLf) is a sialoglycoprotein belonging to the transferrin family, and it plays an important role in immune regulation. It also shows toxicity against cancer cells and pathogenic microorganisms including bacteria, fungi, and virus. The purpose of this study is to assess the roles of the sialylated glycans on bLf against IAV. To this end, bLf were first treated with sodium periodate to destroy its sialylated glycans. Then, the binding activity of native or desialylated bLf with various IAV was assessed by blotting assay. Finally, their ability to inhibit IAV attachment to host cells was analyzed in vitro. Our result showed that the sialylated glycans on bLf were almost completely destroyed by sodium periodate treatment. Furthermore, the binding activity of desialylated bLf to IAV and the ability to inhibit IAV mimics binding to MDCK cells were significantly reduced compared to that of native bLf. These results demonstrated that the sialylated glycans on bLf could serve as competitive substrates to block IAV attachment to host cells during the early stages of viral infection. Our findings make an important contribute for the fully understanding of the mechanism of bLf in the prevention of IAV infections and their possible applications in antiviral infection.

Gene co-expression network analysis of the heat-responsive core transcriptome identifies hub genes in Brassica rapa.

MAIN CONCLUSION: Gene co-expression network analysis of the heat-responsive core transcriptome in two contrasting Brassica rapa accessions reveals the main metabolic pathways, key modules and hub genes, are involved in long-term heat stress. Brassica rapa is a widely cultivated and economically important vegetable in Asia. High temperature is a common stress that severely impacts leaf head formation in B. rapa, resulting in reduced quality and production. The purpose of this study was thus to identify candidate heat tolerance genes by comparative transcriptome analysis of two contrasting B. rapa accessions in response to long-term heat stress. Two B. rapa accessions, '268' and '334', which showed significant differences in heat tolerance, were used for RNA sequencing analysis. We identified a total of 11,055 and 8921 differentially expressed genes (DEGs) in '268' and '334', respectively. Functional enrichment analyses of all of the identified DEGs, together with the genes identified from weighted gene co-expression network analyses (WGCNA), revealed that the autophagy pathway, glutathione metabolism, and ribosome biogenesis in eukaryotes were significantly up-regulated, whereas photosynthesis was down-regulated, in the heat resistance of B. rapa '268'. Furthermore, when B. rapa '334' was subjected to long-term high-temperature stress, heat stress caused significant changes in the expression of certain functional genes linked to protein processing in the endoplasmic reticulum and plant hormone signal transduction pathways. Autophagy-related genes might have been induced by persistent heat stress and remained high during recovery. Several hub genes like HSP17.6, HSP17.6B, HSP70-8, CLPB1, PAP1, PYR1, ADC2, and GSTF11 were discussed in this study, which may be potential candidates for further analyses of the response to long-term heat stress. These results should help elucidate the molecular mechanisms of heat stress adaptation in B. rapa.

[Research on gingival healing situation after stage II surgery of dental implantation for periodontitis patients].

OBJECTIVE: This study aimed to investigate the duration of gingival healing after the stage II surgery of dental implantation for periodontitis patients and to provide clinical guidelines for implant restoration. METHODS: Twenty-nine periodontitis patients who had implantation surgery and achieved osseointegration were operated with stage II surgery (a total of 60 pieces of implants). The height of buccal gingival of each implant was measured twice after the stage II surgery. All implants were measured at the lowest point ofbuccal gingival after one week. The implants were randomly divided into four groups according to the schedule of the next test time: group one at one week from the initial test point, group two at two weeks, group three at three weeks, and group four at four weeks. Each group includes 15 pieces of implants. The amount of the buccal gingival change in each group between the second and first tests was determined, and the data were analyzed statistically. RESULTS: The amount of gingival change of groups one, two, three, and four was (-0.25 +/- 0.66), (-0.04 +/- 0.52), (-0.70 +/- 0.77), and (-0.74 +/- 1.09) mm, respectively. No significant difference was observed between groups one and two in terms of the amount of gingival changes (P > 0.05). However, a significant difference was found between groups two and three (P < 0.05), and the amount of gingival recession was 0.66 mm. No significant difference was found between groups three and four (P > 0.05), and the gingival achieved stability. CONCLUSION: The gingival recession achieves stability at the fourth week (after 28 d) after stage II surgery. At this time, the implant can be restored, and the abutment can be selected according to the amount of gingival change of the periodontitis patient.

[Dynamics of active organic carbon in a paddy soil under different rice farming modes].

A field experiment was conducted to study the dynamics of dissolved organic carbon (DOC), readily oxidizable organic carbon (ROC), and microbial biomass carbon (MBC) in a paddy soil under integrated rice-duck farming (RD), intermittent irrigation (RW), and conventional flooded irrigation (CK), the three rice farming modes typical in southern China. Under these three farming modes, the soil DOC and MBC contents reached the highest during the period from rice booting to heading, while the soil ROC content had less change during the whole rice growth period. Two-factor variance analysis showed that soil MBC was greatly affected by rice growth stage, soil DOC was greatly affected by rice growth stage and farming mode, and soil ROC was mainly affected by farming mode. Comparing with CK, RD significantly increased the soil DOC and ROC contents and their availability, while RW significantly decreased the soil DOC content and its availability but increased the soil ROC content and its availability. No significant differences were observed in the soil MBC and microbial quotient among RD, RW, and CK.