CottonMD: a multi-omics database for cotton biological study.

Cotton is an important economic crop, and many loci for important traits have been identified, but it remains challenging and time-consuming to identify candidate or causal genes/variants and clarify their roles in phenotype formation and regulation. Here, we first collected and integrated the multi-omics datasets including 25 genomes, transcriptomes in 76 tissue samples, epigenome data of five species and metabolome data of 768 metabolites from four tissues, and genetic variation, trait and transcriptome datasets from 4180 cotton accessions. Then, a cotton multi-omics database (CottonMD, http://yanglab.hzau.edu.cn/CottonMD/) was constructed. In CottonMD, multiple statistical methods were applied to identify the associations between variations and phenotypes, and many easy-to-use analysis tools were provided to help researchers quickly acquire the related omics information and perform multi-omics data analysis. Two case studies demonstrated the power of CottonMD for identifying and analyzing the candidate genes, as well as the great potential of integrating multi-omics data for cotton genetic breeding and functional genomics research.

Interaction between phenylpropane metabolism and oil accumulation in the developing seed of Brassica napus revealed by high temporal-resolution transcriptomes.

BACKGROUND: Brassica napus is an important oilseed crop providing high-quality vegetable oils for human consumption and non-food applications. However, the regulation between embryo and seed coat for the synthesis of oil and phenylpropanoid compounds remains largely unclear. RESULTS: Here, we analyzed the transcriptomes in developing seeds at 2-day intervals from 14 days after flowering (DAF) to 64 DAF. The 26 high-resolution time-course transcriptomes are clearly clustered into five distinct groups from stage I to stage V. A total of 2217 genes including 136 transcription factors, are specifically expressed in the seed and show high temporal specificity by being expressed only at certain stages of seed development. Furthermore, we analyzed the co-expression networks during seed development, which mainly included master regulatory transcription factors, lipid, and phenylpropane metabolism genes. The results show that the phenylpropane pathway is prominent during seed development, and the key enzymes in the phenylpropane metabolic pathway, including TT5, BAN, and the transporter TT19, were directly or indirectly related to many key enzymes and transcription factors involved in oil accumulation. We identified candidate genes that may regulate seed oil content based on the co-expression network analysis combined with correlation analysis of the gene expression with seed oil content and seed coat content. CONCLUSIONS: Overall, these results reveal the transcriptional regulation between lipid and phenylpropane accumulation during B. napus seed development. The established co-expression networks and predicted key factors provide important resources for future studies to reveal the genetic control of oil accumulation in B. napus seeds.

Classification and characterization of facial asymmetry in adult patients with skeletal Class III malocclusion using cluster analysis.

INTRODUCTION: This study aimed to analyze the comprehensive maxillofacial features of patients with skeletal Class III malocclusion and facial asymmetry to develop a classification system for diagnosis and surgical planning. METHODS: A total of 161 adult patients were included, with 121 patients in the asymmetry group (menton deviation >2 mm) and 40 patients in the symmetry group (menton deviation ≤2 mm). Twenty-eight variables were determined, including transverse translation, roll and yaw of each facial unit, transverse width, mandibular morphology, and transverse dental compensation. Principal component (PC) analysis was conducted to extract PCs, and cluster analysis was performed using these components to classify the asymmetry group. A decision tree was constructed on the basis of the clustering results. RESULTS: Six PCs were extracted, explaining 80.622% of the data variability. The asymmetry group was classified into 4 subgroups: (1) atypical type (15.7%) showed an opposite roll direction of maxillary dentition than of menton deviation; (2) compound type (34.71%) demonstrated significant ramus height differences, maxillary roll, and mandibular roll and yaw; (3) mandibular yaw type (44.63%) showed slight mandibular yaw without mandibular morphology asymmetry; and (4) maxillary-shift type (4.96%) shared similarities with the compound type but showed significant maxillary translation. The classification and regression tree model achieved a prediction accuracy of up to 85.11%. CONCLUSIONS: This study identified 4 distinct phenotypes using cluster analysis and proposed tailored treatment recommendations on the basis of their specific characteristics. The classification results emphasized the importance of spatial displacement features, especially mandibular yaw, in diagnosing facial asymmetry. The established classification and regression tree model enables clinicians to identify patients conveniently.

Right, but not left, posterior superior temporal gyrus is causally involved in vocal feedback control.

The posterior superior temporal gyrus (pSTG) has been implicated in the integration of auditory feedback and motor system for controlling vocal production. However, the question as to whether and how the pSTG is causally involved in vocal feedback control is currently unclear. To this end, the present study selectively stimulated the left or right pSTG with continuous theta burst stimulation (c-TBS) in healthy participants, then used event-related potentials to investigate neurobehavioral changes in response to altered auditory feedback during vocal pitch regulation. The results showed that, compared to control (vertex) stimulation, c-TBS over the right pSTG led to smaller vocal compensations for pitch perturbations accompanied by smaller cortical N1 and larger P2 responses. Enhanced P2 responses received contributions from the right-lateralized temporal and parietal regions as well as the insula, and were significantly correlated with suppressed vocal compensations. Surprisingly, these effects were not found when comparing c-TBS over the left pSTG with control stimulation. Our findings provide evidence, for the first time, that supports a causal relationship between right, but not left, pSTG and auditory-motor integration for vocal pitch regulation. This lends support to a right-lateralized contribution of the pSTG in not only the bottom-up detection of vocal feedback errors but also the involvement of driving motor commands for error correction in a top-down manner.

Comparative transcriptome profiling reveals the multiple levels of crosstalk in phytohormone networks in Brassica napus.

Plant hormones are the intrinsic factors that control plant development. The integration of different phytohormone pathways in a complex network of synergistic, antagonistic and additive interactions has been elucidated in model plants. However, the systemic level of transcriptional responses to hormone crosstalk in Brassica napus is largely unknown. Here, we present an in-depth temporal-resolution study of the transcriptomes of the seven hormones in B. napus seedlings. Differentially expressed gene analysis revealed few common target genes that co-regulated (up- and down-regulated) by seven hormones; instead, different hormones appear to regulate distinct members of protein families. We then constructed the regulatory networks between the seven hormones side by side, which allowed us to identify key genes and transcription factors that regulate the hormone crosstalk in B. napus. Using this dataset, we uncovered a novel crosstalk between gibberellin and cytokinin in which cytokinin homeostasis was mediated by RGA-related CKXs expression. Moreover, the modulation of gibberellin metabolism by the identified key transcription factors was confirmed in B. napus. Furthermore, all data were available online from http://yanglab.hzau.edu.cn/BnTIR/hormone. Our study reveals an integrated hormone crosstalk network in Brassica napus, which also provides a versatile resource for future hormone studies in plant species.

High-throughput phenotyping-based quantitative trait loci mapping reveals the genetic architecture of the salt stress tolerance of Brassica napus.

Salt stress is a major limiting factor that severely affects the survival and growth of crops. It is important to understand the salt stress tolerance ability of Brassica napus and explore the underlying related genetic resources. We used a high-throughput phenotyping platform to quantify 2111 image-based traits (i-traits) of a natural population under three different salt stress conditions and an intervarietal substitution line (ISL) population under nine different stress conditions to monitor and evaluate the salt stress tolerance of B. napus over time. We finally identified 928 high-quality i-traits associated with the salt stress tolerance of B. napus. Moreover, we mapped the salt stress-related loci in the natural population via a genome-wide association study and performed a linkage analysis associated with the ISL population, respectively. These results revealed 234 candidate genes associated with salt stress response, and two novel candidate genes, BnCKX5 and BnERF3, were experimentally verified to regulate the salt stress tolerance of B. napus. This study demonstrates the feasibility of using high-throughput phenotyping-based quantitative trait loci mapping to accurately and comprehensively quantify i-traits associated with B. napus. The mapped loci could be used for genomics-assisted breeding to genetically improve the salt stress tolerance of B. napus.

Evidence of solvent-mediated proton transfer during H2O2 activation in titanosilicate-catalyzed oxidation systems.

The catalytic performance of titanosilicates involving hydrogen peroxide (H2O2) as the oxidant is strongly influenced by the solvents. Until now, there is still a lack of a universal principle that can guide the choice of a solvent. Herein, the kinetics of H2O2 activation catalyzed by various titanosilicates in different solvents is investigated, and an isokinetic compensation effect is concluded. This indicates that the solvent participates in the H2O2 activation process for the formation of a Ti-OOH species. Additionally, the results of isotopically labeled infrared spectra preliminarily confirm that the solvent acts as the mediator to promote the proton transfer during the H2O2 activation process. The catalytic activities of a series of TS-1 catalysts toward 1-hexene epoxidation are compared, which include Ti(OSi)3OH species with a range of densities but a constant total Ti content. This reveals that the solvent effect is closely related to the Ti active sites of these TS-1 catalysts. Based on these results, a principle for the rational choice of solvent for this catalytic process is proposed. ROH is found to be the mediator for Ti(OSi)4 sites, and methanol, which has a strong proton-donating ability, is the best solvent for these sites. However, for the Ti(OSi)3OH sites, water (H2O) is the mediator, and a weaker hydrogen bonding between H2O molecules promotes proton transfer more effectively. The solvent influences the catalytic performance by perturbing the hydrogen bonds between the H2O molecules, and aprotic acetonitrile, which has a strong ability to break the hydrogen bonding network between H2O molecules, is the best solvent for Ti(OSi)3OH sites. This study provides experimental evidence that the solvent promotes the catalytic performance of titanosilicates by assisting the proton transfer during the catalytic H2O2 activation process, which will pave the way toward the rational choice of solvent for the titanosilicate-catalyzed oxidation systems.

Calling large indels in 1047 Arabidopsis with IndelEnsembler.

Large indels greatly impact the observable phenotypes in different organisms including plants and human. Hence, extracting large indels with high precision and sensitivity is important. Here, we developed IndelEnsembler to detect large indels in 1047 Arabidopsis whole-genome sequencing data. IndelEnsembler identified 34 093 deletions, 12 913 tandem duplications and 9773 insertions. Our large indel dataset was more comprehensive and accurate compared with the previous dataset of AthCNV (1). We captured nearly twice of the ground truth deletions and on average 27% more ground truth duplications compared with AthCNV, though our dataset has less number of large indels compared with AthCNV. Our large indels were positively correlated with transposon elements across the Arabidopsis genome. The non-homologous recombination events were the major formation mechanism of deletions in Arabidopsis genome. The Neighbor joining (NJ) tree constructed based on IndelEnsembler's deletions clearly divided the geographic subgroups of 1047 Arabidopsis. More importantly, our large indels represent a previously unassessed source of genetic variation. Approximately 49% of the deletions have low linkage disequilibrium (LD) with surrounding single nucleotide polymorphisms. Some of them could affect trait performance. For instance, using deletion-based genome-wide association study (DEL-GWAS), the accessions containing a 182-bp deletion in AT1G11520 had delayed flowering time and all accessions in north Sweden had the 182-bp deletion. We also found the accessions with 65-bp deletion in the first exon of AT4G00650 (FRI) flowered earlier than those without it. These two deletions cannot be detected in AthCNV and, interestingly, they do not co-occur in any Arabidopsis thaliana accession. By SNP-GWAS, surrounding SNPs of these two deletions do not correlate with flowering time. This example demonstrated that existing large indel datasets miss phenotypic variations and our large indel dataset filled in the gap.

Risk, Prognostic Factors and Nomograms for Bone Metastasis in Young Females with Breast Cancer: A Large Cohort Retrospective Study.

Purpose: To determine the incidence of bone metastasis (BM) in young female patients with breast cancer (BC) and develop 2 robust nomograms for BM in young female patients with BC. Methods: We searched and downloaded the data from young (age ≤40 years) female patients with bone cancer from the Surveillance, Epidemiology, and End Results (SEER) database between 2010 and 2015. Univariate and multivariate analyses were performed to screen the potential diagnostic variables and prognostic factors for BM. The diagnostic and prognostic nomograms were generated and evaluated by the area under the receiver operating characteristic (ROC) curve (AUC), calibration curves, and decision curve analysis (DCA). Results: A total of 13 347 young female patients with BC were identified; of these, 462 were initially diagnosed as having BM. The independent risk factors for BM in young female patients with BC were tumor size, BC subtype, American Joint Committee on Cancer (AJCC) T stage, AJCC N stage, age and marital status. The independent prognostic factors in these patients were tumor size, subtype, surgery performed, lung metastasis, liver metastasis and brain metastasis. The AUC values of the diagnostic nomogram were 0.803 (95% CI; 0.795-0.811) and 0.813 (95% CI; 0.800-0.825) in the training and validation cohorts, respectively. The time-dependent AUC values of prognostic nomogram were 0.850, 0.853, and 0.824 at 2, 3 and 4 years in the training cohort, and also >0.700 in the validation cohort. For both nomograms, the discrimination was higher than all independent variables. Calibration curve and decision curve analysis (DCA) indicated that both nomograms had favorable calibration and clinical utilization. Finally, a risk stratification system was generated and the 3 risk subgroups showed significantly distinct prognoses. Conclusions: A total of 2 nomograms were developed to assess the risk for and in prognosis of young female patients with BC with BM (BCBM).

Neuroprotective Effects of Savinin on LPS-Induced Neuroinflammation In Vivo via Regulating MAPK/NF-κB Pathway and NLRP3 Inflammasome Activation.

The traditional herb Eleutherococcus henryi Oliv. is commonly used to treat inflammatory conditions including rheumatism, arthritis, and hepatitis, as well as mental fatigue and amnesia, according to traditional Chinese medicine (TCM) theory. Savinin is a natural lignan obtained from the roots of E. henryi. The present study was undertaken to determine whether savinin can relieve LPS-induced neuroinflammation and if so, what the mechanism is. Groups of male C57BL/6 mice were administered savinin (5, 10, 20 mg/kg) and DEX (10 mg/kg) by gavage once daily for a continuous 7 days. On the 5th day of continuous pre-administration, LPS (2.5 mg/kg) was injected into the lateral ventricles of the mice for modeling 48 h. We found that treatment with savinin decreased the levels of neuroinflammatory cytokines and histopathological alterations dramatically. Consequently, it improved the LPS-induced neuroinflammatory response in mice. Furthermore, savinin inhibited the up-regulated expression of related proteins in the activated MAPK/NF-κB and NLRP3 inflammasome signaling pathways caused by LPS. Docking studies demonstrated the binding of savinin to three receptors (MAPK, NF-κB and NLRP3) using a well-fitting mode. These findings suggest that savinin may suppress neuroinflammation induced by LPS in vivo via modulating MAPK/NF-κB and NLRP3 signaling pathways.

Three-dimensional (3D) quantitative evaluation of the morphological changes of the upper anterior alveolar bone after retraction of a maxillary incisor.

BACKGROUND: The purpose of this study was to assess morphological changes of the upper anterior alveolus after retraction of a maxillary incisor by applying three-dimensional (3D) superimposition of pretreatment (T1) and posttreatment (T2) cone-beam computed tomography (CBCT) data. METHODS: The study group was comprised of 28 patients with skeletal Class II malocclusion who underwent incisor retraction. CBCT data were acquired before (T1) and after (T2) orthodontic treatment. Labial and palatal alveolar thickness were assessed at the crestal, midroot and apical levels of the retracted incisors. Following three-dimensional (3D) cranial base superimposition, we performed surface modeling and inner remodeling of the labial and palatal alveolar cortex of the maxillary incisors. Paired t-tests were used to compare T0 and T1 bone thickness and volume measurements. Comparisons between labial and palatal surface modeling, inner remodeling and outer surface modeling were performed with paired t-tests in SPSS 20.0 version. RESULTS: We observed controlled tipping retraction of the upper incisor. After treatment, the alveolar thickness on the labial sides increased and the palatal alveolar thickness decreased. The labial cortex showed a wider range of modeling area with a larger bending height and a smaller bending angle than the palatal side. The extent of inner remodeling was more prominent than the outer surface on both the labial and palatal sides. CONCLUSIONS: Adaptive alveolar surface modeling occurred in response to incisor tipping retraction on both the lingual and labial sides although these changes occurred in an uncoordinated manner. Tipping retraction of the maxillary incisors led to a reduction in alveolar volume.

A novel maxillary transverse deficiency diagnostic method based on ideal teeth position.

BACKGROUND: This study proposed a novel maxillary transverse deficiency diagnostic method and evaluated the skeletal Class I and the mild skeletal Class III groups. METHODS: Pre-treatment data from 30 mild skeletal Class III and 30 skeletal Class I patients were collected and uploaded to the Emeiqi Case Management System to design the ideal teeth positions. On these positions, the first bi-molars width was measured at the central fossa and center resistance, the maxillary first bi-premolars width was measured at the central fossa, and the mandibular first bi-premolars width was measured at the distal contact point by Mimics, then width differences of two groups were calculated respectively. RESULTS: At ideal teeth positions, there was no statistically significant difference in the maxillomandibular width in the premolar area between the two groups, but there was in the molar area, and this difference was caused by the difference in mandible width between the two groups. CONCLUSIONS: We proposed a new transverse diagnostic method and found that even the Class I group was not quite up to standard in the molar area on ideal teeth positions, and the Class III group had more severe maxillary transverse deficiency than the Class I group. Meanwhile, the maxillary transverse deficiency in the Class III group was mainly caused by the larger width of the mandible.

A causal link between left supplementary motor area and auditory-motor control of vocal production: Evidence by continuous theta burst stimulation.

The supplementary motor area (SMA) has been implicated in the feedforward control of speech production. Whether this region is involved in speech motor control through auditory feedback, however, remains uncertain. The present event-related potential (ERP) study examined the role of the left SMA in vocal pitch regulation in a causal manner by combining auditory feedback manipulations and neuronavigated continuous theta bust stimulation (c-TBS). After receiving c-TBS over the left SMA or the control site (vertex), twenty young adults vocalized the vowel sound /u/ while hearing their voice unexpectedly pitch-shifted -50 or -200 cents. Compared to the control stimulation, c-TBS over the left SMA led to decreased vocal compensations for pitch perturbations of -50 and -200 cents. A significant decrease of N1 and P2 responses to -200 cents perturbations was also found when comparing active and control stimulation. Major neural generators of decreased P2 responses included the right-lateralized superior and middle temporal gyrus and angular gyrus. Notably, a significant correlation was found between active-control differences in the vocal compensation and P2 responses for the -200 cents perturbations. These findings provide neurobehavioral evidence for a causal link between the left SMA and auditory-motor integration for vocal pitch regulation, suggesting that the left SMA receives auditory feedback information and mediates vocal compensations for feedback errors in a bottom-up manner.

Combined analysis of receptor expression reflects inter-and intra-tumor heterogeneity in HR+/HER2+ breast cancer.

BACKGROUND: Hormone receptor-positive and human epidermal growth factor receptor 2-positive (HR+/HER2+ breast cancer comprise approximately 5-10% of all invasive breast cancers. However, the lack of knowledge regarding the complexity of tumor heterogeneity in HR+/HER2+ disease remains a barrier to more accurate therapies. This study aimed to describe the tumor heterogeneity of HR+/HER2+ breast cancer and to establish a novel indicator to identify the HER2-enriched subtype in patients with HR+/HER2+ breast cancer. METHODS: First of all, a comprehensive analysis was performed on HR+/HER2+ breast cancer samples from the TCGA (n = 141) and METABRIC (n = 104) databases. We determined the distribution of PAM50 intrinsic subtypes within the two cohorts and compared the somatic mutational profile and RNA expression features between HER2-enriched and non-HER2-enriched subtypes. From this, we constructed a novel marker termed rH/E, which was calculated as ERBB2 expression quantity/(ESR1 expression quantity + 1). Secondly, we performed multiplex immunofluorescence (mIF) to evaluate HER2 and estrogen receptor (ER) expression simultaneously in the third cohort, enrolling 43 cases of early HR+/HER2+ breast cancer from Cancer Hospital, Chinese Academy of Medical Sciences (CAMS). When using mIF, rH/E was adjusted to prH/E, which was calculated as HER2-positive cells%/(ER-positive cells + 1)%. RESULTS: All four main intrinsic subtypes were identified in HR+/HER2+ breast cancer, of which the luminal B subtype was the most common, followed by the HER2-enriched and luminal A subtypes. Significantly increased TP53 and ERBB3 and decreased PIK3CA somatic mutation frequency were observed in the HER2-enriched subtype compared with the non-HER2-enriched subtype. In addition, the HER2-enriched subtype was characterized by significantly higher ERBB2 and lower ESR1 expression. We then constructed a marker termed rH/E to reflect the relative expression of ERBB2 to ESR1 in each patient. rH/E discriminates the HER2-enriched subtype from the better than the expression of ERBB2 or ESR1 alone. In the CAMS cohort, we observed four subtypes of tumor cells: ER+/HER2-, ER+/HER2+, ER-/HER2+, and ER-/HER2-. Tumor cell diversity was common, with 86% of patients having all four subtypes of tumor cells. Moreover, prH/E showed a significant prognostic association in the CAMS cohort. CONCLUSIONS: This study furthers our understanding of the complexity of tumor heterogeneity in HR+/HER2+ breast cancer, and suggests that the combined analysis of ERBB2 and ESR1 expression may contribute to identifying patients with specific subtypes in this population.

R gene triplication confers European fodder turnip with improved clubroot resistance.

Clubroot is one of the most important diseases for many important cruciferous vegetables and oilseed crops worldwide. Different clubroot resistance (CR) loci have been identified from only limited species in Brassica, making it difficult to compare and utilize these loci. European fodder turnip ECD04 is considered one of the most valuable resources for CR breeding. To explore the genetic and evolutionary basis of CR in ECD04, we sequenced the genome of ECD04 using de novo assembly and identified 978 candidate R genes. Subsequently, the 28 published CR loci were physically mapped to 15 loci in the ECD04 genome, including 62 candidate CR genes. Among them, two CR genes, CRA3.7.1 and CRA8.2.4, were functionally validated. Phylogenetic analysis revealed that CRA3.7.1 and CRA8.2.4 originated from a common ancestor before the whole-genome triplication (WGT) event. In clubroot susceptible Brassica species, CR-gene homologues were affected by transposable element (TE) insertion, resulting in the loss of CR function. It can be concluded that the current functional CR genes in Brassica rapa and non-functional CR genes in other Brassica species were derived from a common ancestral gene before WGT. Finally, a hypothesis for CR gene evolution is proposed for further discussion.

Thromboembolic events in Janus kinase inhibitors: A pharmacovigilance study from 2012 to 2021 based on the Food and Drug Administration's Adverse Event Reporting System.

AIMS: As a new type of drug developed rapidly in recent years, Janus kinase inhibitors (JAKinibs) have caused controversy due to possible adverse reactions of thromboembolism. The aim of this study was to analyse and evaluate the association between thromboembolic events and the use of JAKinibs, on the base of the latest data in the Food and Drug Administration's Adverse Event Reporting System. METHODS: A disproportionality analysis was conducted, utilizing data from 1 January 2012 to 30 September 2021 in the FAERS. For each drug-adverse event pair, reporting odds ratio (ROR) and information components (IC) were calculated. RESULTS: A total of 15 positive safety signals were detected within the FAERS: ruxolitinib was significantly associated with portal vein thrombosis (ROR025 = 3.49, IC025 = 1.50); tofacitinib immediate release with pulmonary embolism (ROR025 = 2.09, IC025 = 1.02) and thrombosis (ROR025 = 1.15, IC025 = 0.18); tofacitinib extended release with pulmonary embolism (ROR025 = 1.27, IC025 = 0.26) and thrombosis (ROR025 = 1.29, IC025 = 0.33); baricitinib with deep vein thrombosis (ROR025 = 8.27, IC025 = 3.00), portal vein thrombosis (ROR025 = 1.97, IC025 = 0.63), pulmonary embolism (ROR025 = 7.90, IC025 = 2.94), thrombosis (ROR025 = 2.04, IC025 = 0.93) and venous thrombosis (ROR025 = 2.15, IC025 = 0.81); upadacitinib with pulmonary embolism (ROR025 = 1.25, IC025 = 0.25), pulmonary thrombosis (ROR025 = 5.32, IC025 = 2.33) and thrombosis (ROR025 = 2.72, IC025 = 1.39); and filgotinib with pulmonary embolism (ROR025 = 4.83, IC025 = 2.10). In the analysis of the time to onset of thromboembolic events, no obviously recognizable pattern was found. Several safety signals with embolic and thrombotic events (Standardised MedDRA Query) were found in the study. CONCLUSION: This pharmacovigilance study covered 8 types of JAKinib that are already on the market, and provided new safety signals based on past safety information. Some of these signals still need more medical evidence.

Cellular communication network factor 1 interlinks autophagy and ERK signaling to promote osteogenesis of periodontal ligament stem cells.

OBJECTIVES: To investigate the effects of cellular communication network factor 1 (CCN1), a critical matricellular protein, on alveolar bone regeneration, and to elucidate the underlying molecular mechanism. BACKGROUND: In the process of orthodontic tooth movement, bone deposition on the tension side of human periodontal ligament stem cells (hPDLSCs) ensured high efficiency and long-term stability of the treatment. The matricellular protein CCN1 is responsive to mechanical stimulation, exhibiting important tasks in bone homoeostasis. However, the role and mechanism of CCN1 on alveolar bone remodeling of hPDLSCs remains unclear. METHODS: The expression and distribution of CCN1 in rat periodontal ligament were detected by immunofluorescence staining and immunohistochemical staining. ELISA verified the secretion of CCN1 triggered by stretch loading. To examine the mineralization ability of hPDLSCs induced by CCN1, Western blotting, qRT-PCR, ARS, and ALP staining were performed. CCK-8 and cell migration assay were performed to detect the cell proliferation rate and the wound healing. PI3K/Akt, MAPK, and autophagy activation were examined via Western blotting and immunofluorescence. RESULTS: Mechanical stimuli induced the release of CCN1 into extracellular environment by hPDLSCs. Knockdown of CCN1 attenuated the osteogenesis of hPDLSCs while rhCCN1 enhanced the expression of Runx2, Col 1, ALPL, and promoted the mineralization nodule formation. CCN1 activated PI3K/Akt and ERK signaling, and blockage of PI3K/Akt signaling reversed the accelerated cell migration triggered by CCN1. The enhanced osteogenesis induced by CCN1 was abolished by ERK signaling inhibitor PD98059 or autophagy inhibitor 3-MA. Further investigation demonstrated PD98059 abrogated the activation of autophagy. CONCLUSION: This study demonstrated that CCN1 promotes osteogenesis in hPDLSCs via autophagy and MAPK/ERK pathway.

Evaluating the role of Ubiquitin D gene expression in types of leukemia.

Using animal models to develop new treatments is essential, especially in diseases like cancer. In this study, we induced leukemia by intravenous injection of cancer cells (BCL1 cell line) and the examination of cell markers in the animal's blood to study the changes in the expression of the UBD gene as a biomarker for diagnosing and examining the progress of the disease. For this purpose, five million BCL-1 cells were injected into the tail vein of BALBIe mice of the same breed. Fifty mice were killed after four weeks, and we examined peripheral blood cells and histological changes. Then RNA of the samples was extracted, and cDNA synthesis was done with the help of MMuLV enzyme, Oligo dT, and Random hexamer primers. Specific primers for UBD were designed using Primer Express software, and the expression level of the UBD gene was measured by the method. The results showed that in the CML group, the lowest expression level was 1.70 times, and in the ALL group, the highest expression level was 7.97 times compared to the control group. The average increase in UBD gene expression was 3.21 times in the CLL group and 4.94 times in the AML group. The UBD gene can be further investigated so that it may be used as a proposed biomarker for the diagnosis of leukemia. Therefore, the evaluation of the expression level of this gene can be used to diagnose leukemia. However, more studies than the currently applied methods are needed in cancer diagnosis, which has many errors compared to the technique used in this study, and to prove its accuracy and sensitivity.

Nrf2 activation is involved in osteogenic differentiation of periodontal ligament stem cells under cyclic mechanical stretch.

During orthodontic treatment, mechanical stretch serves a crucial function in osteogenic differentiation of periodontal ligament stem cells (PDLSCs). Up-regulated reactive oxygen species (ROS) level is a result of cyclic mechanical stretch in many cell types. Nuclear factor erythroid-2-related factor-2 (Nrf2) is a master regulator in various antioxidants expression. However, it is not known whether cyclic mechanical stretch could induce the ROS generation in PDLSCs and whether Nrf2 participated in this process. The present study was aimed to investigate the role of Nrf2 in PDLSCs under cyclic mechanical stretch. Our results showed that cyclic mechanical stretch increased ROS level and the nuclear accumulation of Nrf2 during osteoblast differentiation. Knocking down Nrf2 by siRNA transfection increased ROS formation and suppressed osteogenic differentiation in PDLSCs. T-BHQ, a Nrf2 activator, promoted the osteogenic differentiation in PDLSCs under cyclic mechanical stretch, and improved the microstructure of alveolar bone during orthodontic tooth movement in rats by employing micro-CT system. Taken together, Nrf2 activation was involved in osteogenic differentiation under cyclic mechanical stretch in PDLSCs. T-BHQ could promote the osteogenic differentiation in vitro and in vivo, suggesting a promising option for the remodeling of the alveolar bone during orthodontic tooth movement.

A New Method for Anterior Boundary Demarcation of the Nasopharynx in Three-Dimensional Analysis.

ABSTRACT: Three-dimensional (3D) measurements of the upper airway have been extensively applied and researched, but the division of the airway is carried out in various ways, especially when demarcating the anterior boundary of the nasopharynx. The present study was to propose a new method based on the anatomical definition for the anterior boundary demarcation of the nasopharynx used in three-dimensional analysis. Twenty computed tomography scans (age 9.5 ±â€Š2.5 years, 11 males, and 9 females) of head and neck were randomly selected and transferred to Materialism's interactive medical image control system 19.0 for segmentation of the nasopharynx. Precise localization of the reference points that determining the nasopharyngeal anterior and inferior boundaries was managed by recording their coordinates in the interface of the software. The area of the anterior and the inferior boundaries, and the volume of the nasopharynx were measured and repeated with a 2-week interval for the consistency test. Both the interobserver reliability as well as the intra-observer reliability were very high (intraclass correlation coefficients, 0.985-0.997). Paired t test showed no significant difference between the first and the second examinations. This new simple method proposed for demarcation of the nasopharyngeal anterior boundary was based on the innate anatomical boundary, which was statistically reliable, technically convenient, and clinically reasonable.