Inhibition of soluble epoxide hydrolase enhances the dentin-pulp complex regeneration mediated by crosstalk between vascular endothelial cells and dental pulp stem cells.

BACKGROUND: Revascularization and restoration of normal pulp-dentin complex are important for tissue-engineered pulp regeneration. Recently, a unique periodontal tip-like endothelial cells subtype (POTCs) specialized to dentinogenesis was identified. We have confirmed that TPPU, a soluble epoxide hydrolase (sEH) inhibitor targeting epoxyeicosatrienoic acids (EETs) metabolism, promotes bone growth and regeneration by angiogenesis and osteogenesis coupling. We hypothesized that TPPU could also promote revascularization and induce POTCs to contribute to pulp-dentin complex regeneration. Here, we in vitro and in vivo characterized the potential effect of TPPU on the coupling of angiogenesis and odontogenesis and investigated the relevant mechanism, providing new ideas for pulp-dentin regeneration by targeting sEH. METHODS: In vitro effects of TPPU on the proliferation, migration, and angiogenesis of dental pulp stem cells (DPSCs), human umbilical vein endothelial cells (HUVECs) and cocultured DPSCs and HUVECs were detected using cell counting kit 8 (CCK8) assay, wound healing, transwell, tube formation and RT-qPCR. In vivo, Matrigel plug assay was performed to outline the roles of TPPU in revascularization and survival of grafts. Then we characterized the VEGFR2 + POTCs around odontoblast layer in the molar of pups from C57BL/6 female mice gavaged with TPPU. Finally, the root segments with DPSCs mixed with Matrigel were implanted subcutaneously in BALB/c nude mice treated with TPPU and the root grafts were isolated for histological staining. RESULTS: In vitro, TPPU significantly promoted the migration and tube formation capability of cocultured DPSCs and HUVECs. ALP and ARS staining and RT-qPCR showed that TPPU promoted the osteogenic and odontogenic differentiation of cultured cells, treatment with an anti-TGF-ß blocking antibody abrogated this effect. Knockdown of HIF-1α in HUVECs significantly reversed the effect of TPPU on the expression of angiogenesis, osteogenesis and odontogenesis-related genes in cocultured cells. Matrigel plug assay showed that TPPU increased VEGF/VEGFR2-expressed cells in transplanted grafts. TPPU contributed to angiogenic-odontogenic coupling featured by increased VEGFR2 + POTCs and odontoblast maturation during early dentinogenesis in molar of newborn pups from C57BL/6 female mice gavaged with TPPU. TPPU induced more dental pulp-like tissue with more vessels and collagen fibers in transplanted root segment. CONCLUSIONS: TPPU promotes revascularization of dental pulp regeneration by enhancing migration and angiogenesis of HUVECs, and improves odontogenic differentiation of DPSCs by TGF-ß. TPPU boosts the angiogenic-odontogenic coupling by enhancing VEGFR2 + POTCs meditated odontoblast maturation partly via upregulating HIF-1α, which contributes to increasing pulp-dentin complex for tissue-engineered pulp regeneration.

Genetic causal association between rheumatoid arthritis and periodontitis: a bidirectional two-sample Mendelian randomization analysis.

BACKGROUND: A link between rheumatoid arthritis (RA) and periodontitis (PD) has been established. However, their causal relationship remains unclear and the effect of different serotypes of RA on the PD development has not been investigated. This study aims to elucidate the causal association between PD and different serotypes of RA using Mendelian randomization (MR). METHODS: A bidirectional two-sample MR analysis was performed using available large-scale genome-wide association studies statistics. The inverse-variance weighted (IVW) or multiplicative random-effects IVW was used to determine causality, depending on the heterogeneity of instrumental variables. Additional sensitivity analyses were also performed. RESULTS: The forward MR analysis identified that seropositive RA (odds ratio (OR), 1.26; 95% confidence interval (CI), 1.07-1.44; p = 0.0018), but not seronegative RA (OR, 1.01; 95% CI, 0.95-1.06; p = 0.9098), was associated with an increased risk of PD. The reverse MR analysis did not show any significant causal effect of PD on RA, independent of the serotypes. The sensitivity tests (p > 0.05) confirmed the robustness and accuracy of these findings. CONCLUSION: This study revealed that there was a genetic causal effect of seropositive RA on PD, suggesting that this subtype of RA patients may require specific clinical attention to prevent the development of PD.

Does dental caries play a role on the asthma development?-systematic review and meta-analysis.

Asthma and dental caries are the two most common diseases in children, and the relationship between them has been a focus of research. Whether dental caries affects the development of asthma has long been controversial. The aim of this study was to conduct a systematic review of the literature to assess the effect of dental caries on the development of asthma and provide new ideas for the pathogenesis and promoting factors of asthma. For a systematic review and meta-analysis, we systematically searched three databases (PubMed, Web of Science, and Embase) for studies published from database inception to 22 May 2022. We included observational studies that investigated the effect of dental caries on the development of asthma. Studies were critically appraised and a meta-analysis was performed to estimate a pooled effect. From the 845 studies initially identified, 7 of these were included in the meta-analysis. Included studies originated from America (n = 5) and Asia (n = 2). A meta-analysis of data from seven selected studies suggested that dental caries was positively associated with the risk of asthma development (The odds ratio for the pooled effect was 1.06, 95% confidence interval: 1.01, 1.10). In addition, the effect of dental caries on asthma risk varied in different geographic locations according to subgroup analyses. This study suggests that dental caries may affect the development of asthma and emphasizes the importance of increased awareness of dental care and caries prevention in patients with asthma.

Polarization of macrophages in the trigeminal ganglion of rats with pulpitis.

BACKGROUND: Dental pulp tissues are rich in pain-related afferent nerve fibers, which originate from primary sensory neurons in the trigeminal ganglion (TG). The mechanisms of central nervous system (CNS) underlying ectopic pain following peripheral inflammation have been reported that the macrophages as inflammatory and immunologic mediators in the TG play an important role in the process of pulpitis and hyperalgesia. OBJECTIVE(S): To observe the polarization response and dynamic distribution of macrophages in the TG during the development of dental pulp inflammation. METHODS: A rat model of pulpitis was established using complete Freund's adjuvant (CFA). Hematoxylin-eosin (HE), immunohistochemistry (IHC), immunofluorescence (IF), toluidine blue (TB) staining, and RT-qPCR were performed to observe the expression of macrophage-related factors in the TG. RESULTS: The results of IHC staining showed that M2 macrophages labeled with CD206 were observed in the TG of both the control and CFA groups. The statistical analysis indicated that the number of CD206-positive macrophages in the TG increased significantly at 24 h after CFA-induced pulpitis, reached a peak at 2 weeks, and then returned to the normal level after 6 weeks. The ratio of M2/M1 in the CFA groups was significantly lower than that in the control group from 24 to 72 h, and this pattern was reversed at 2 weeks after CFA-induced pulpitis; then, the ratio increased significantly and was maintained at a high level for 4 weeks. RT-qPCR results showed that the expression of IL-10 in the TG increased significantly from 1 to 4 weeks after CFA-induced pulpitis. CONCLUSION: The trend of M2 macrophages was opposite to that of M1 macrophages in the TG during the process of pulpitis induced by CFA, which is consistent with the expression of macrophage-related cytokines. Macrophage polarization in the TG may participate in the neuroinflammation response induced by dental pulpitis.

Role of PI3K in the bone resorption of apical periodontitis.

BACKGROUND: Phosphoinositide 3-kinase (PI3K) is located within cells, and is involved in regulating cell survival, proliferation, apoptosis and angiogenesis. The purpose of this study was to investigate the role of PI3K in the process of bone destruction in apical periodontitis, and provide reference data for the treatment of this disease. METHODS: The relative mRNA expression of PI3K, Acp5 and NFATc1 in the normal human periodontal ligament and in chronic apical periodontitis were analyzed by real-time quantitative polymerase chain reaction (RT-qPCR). A mouse model of apical periodontitis was established by root canal exposure to the oral cavity, and HE staining was used to observe the progress of apical periodontitis. Immunohistochemical staining was used to detect the expression of PI3K and AKT in different stages of apical periodontitis, while enzymatic histochemical staining was used for detection of osteoclasts. An Escherichia coli lipopolysaccharide (LPS)-mediated inflammatory environment was also established at the osteoclast and osteoblast level, and osteoclasts or osteoblasts were treated with the PI3K inhibitor LY294002 to examine the role of PI3K in bone resorption. RESULTS: The expression of PI3K, Acp5 and NFATc1 genes in chronic apical periodontitis sample groups was significantly increased relative to healthy periodontal ligament tissue (P < 0.05). Mouse apical periodontitis was successfully established and bone resorption peaked between 2 and 3 weeks (P < 0.05). The expression of PI3K and Akt increased with the progression of inflammation, and reached a peak at 14 days (P < 0.05). The gene and protein expression of PI3K, TRAP and NFATc1 in osteoclasts were significantly increased (P < 0.05) in the E. coli LPS-mediated inflammatory microenvironment compared to the normal control group. Meanwhile in osteoblasts, the gene and protein expression of PI3K, BMP-2 and Runx2 were significantly reduced (P < 0.05) in the inflammatory microenvironment. With the addition of LY294002, expressions of bone resorption-related factors (TRAP, NFATc1) and bone formation-related factors (BMP-2, Runx2) significantly decreased (P < 0.05). CONCLUSIONS: Under the inflammatory environment induced by LPS, PI3K participates in the occurrence and development of chronic apical periodontitis by regulating the proliferation and differentiation of osteoclasts and osteoblasts.

Increased salivary microvesicles are associated with the prognosis of patients with oral squamous cell carcinoma.

Microvesicles (MVs), which are cell-derived membrane vesicles present in body fluids, are closely associated with the development of malignant tumours. Saliva, one of the most versatile body fluids, is an important source of MVs. However, the association between salivary MVs (SMVs) and oral squamous cell carcinoma (OSCC), which is directly immersed in the salivary milieu, remains unclear. SMVs from 65 patients with OSCC, 21 patients with oral ulcer (OU), and 42 healthy donors were purified, quantified and analysed for their correlations with the clinicopathologic features and prognosis of OSCC patients. The results showed that the level of SMVs was significantly elevated in patients with OSCC compared to healthy donors and OU patients. Meanwhile, the level of SMVs showed close correlations with the lymph node status, and the clinical stage of OSCC patients. Additionally, the ratio of apoptotic to non-apoptotic SMVs was significantly decreased in OSCC patients with higher pathological grade. Consistently, poorer overall survival was observed in patients with lower ratio of apoptotic to non-apoptotic SMVs. In conclusion, the elevated level of SMVs is associated with clinicopathologic features and decreased survival in patients with OSCC, suggesting that SMVs are a potential biomarker and/or regulator of the malignant progression of OSCC.

Elastomeric negative acoustic contrast particles for capture, acoustophoretic transport, and confinement of cells in microfluidic systems.

We present a particle-based method for the immunospecific capture and confinement of cells using acoustic radiation forces. Ultrasonic standing waves in microfluidic systems have previously been used for the continuous focusing of cells in rapid screening and sorting applications. In aqueous fluids, cells typically exhibit positive acoustic contrast and are thus forced toward the pressure nodes of a standing wave. Conversely, elastomeric particles exhibit negative acoustic contrast and travel toward the pressure antinodes. We have developed a class of elastomeric particles that are synthesized in bulk using a simple nucleation and growth process, providing precise control over their size and functional properties. We demonstrate that the biofunctionalization of these particles can allow the capture and transport of cells to the pressure antinodes solely via acoustic radiation forces, which may enable new acoustics-based cell handling techniques such as the washing, labeling, and sorting of cells with minimal preparatory steps.

Nucleation and growth synthesis of siloxane gels to form functional, monodisperse, and acoustically programmable particles.

Nucleation and growth methods offer scalable means of synthesizing colloidal particles with precisely specified size for applications in chemical research, industry, and medicine. These methods have been used to prepare a class of silicone gel particles that display a range of programmable properties and narrow size distributions. The acoustic contrast factor of these particles in water is estimated and can be tuned such that the particles undergo acoustophoresis to either the pressure nodes or antinodes of acoustic standing waves. These particles can be synthesized to display surface functional groups that can be covalently modified for a range of bioanalytical and acoustophoretic sorting applications.

Elastomeric microparticles for acoustic mediated bioseparations.

BACKGROUND: Acoustophoresis has been utilized successfully in applications including cell trapping, focusing, and purification. One current limitation of acoustophoresis for cell sorting is the reliance on the inherent physical properties of cells (e.g., compressibility, density) instead of selecting cells based upon biologically relevant surface-presenting antigens. Introducing an acoustophoretic cell sorting approach that allows biochemical specificity may overcome this limitation, thus advancing the value of acoustophoresis approaches for both the basic research and clinical fields. RESULTS: The results presented herein demonstrate the ability for negative acoustic contrast particles (NACPs) to specifically capture and transport positive acoustic contrast particles (PACPs) to the antinode of an ultrasound standing wave. Emulsification and post curing of pre-polymers, either polydimethylsiloxane (PDMS) or polyvinylmethylsiloxane (PVMS), within aqueous surfactant solution results in the formation of stable NACPs that focus onto pressure antinodes. We used either photochemical reactions with biotin-tetrafluorophenyl azide (biotin-TFPA) or end-functionalization of Pluronic F108 surfactant to biofunctionalize NACPs. These biotinylated NACPs bind specifically to streptavidin polystyrene microparticles (as cell surrogates) and transport them to the pressure antinode within an acoustofluidic chip. CONCLUSION: To the best of our knowledge, this is the first demonstration of using NACPs as carriers for transport of PACPs in an ultrasound standing wave. By using different silicones (i.e., PDMS, PVMS) and curing chemistries, we demonstrate versatility of silicone materials for NACPs and advance the understanding of useful approaches for preparing NACPs. This bioseparation scheme holds potential for applications requiring rapid, continuous separations such as sorting and analysis of cells and biomolecules.

Oral Health Status of Disabled Children and Adolescents in China.

OBJECTIVE: To investigate the dental caries status and periodontal status of disabled children and adolescents aged 0 to 18 years in China and provide suggestions for future policies. METHODS: The cross-sectional survey included disabled children and adolescents aged 0 to 18 years in welfare institutions across 10 provinces in eastern, central and western regions in China, between November 2013 and May 2015. Oral health examination included dental caries status and periodontal status. The mean score for decayed, missing and filled teeth (dmft/ DMFT) due to caries in primary/permanent dentition, gingival bleeding rate and calculus rate were recorded and statistically evaluated. Data analysis was performed using SPSS 24.0 software (IBM, Armonk, NY, USA), and the level of statistical significance was P < 0.05. RESULTS: The mean dmft/DMFT values of for groups aged 0 to 5, 6 to 12 and 13 to 18 years were 1.48 ± 3.00, 2.19 ± 2.94 and 1.78 ± 2.93, respectively. The mean scores for the groups aged 0 to 5 (P < 0.05) and 13 to 18 years (P < 0.01) showed a significant difference among different disability types. A significant difference between sexes was found only in the group aged 0 to 5 years (P < 0.05). Gingival bleeding rates were 13.70% for the group aged 0 to 5 years, 24.81% for the group aged 6 to 12 years and 42.06% for the group aged 13 to 18 years, and calculus rates were 5.48%, 22.41% and 47.62% for the three age groups, respectively. The gingival bleeding rate for all three age groups showed a significant difference between different disability types (P < 0.01), whereas calculus rates a showed significant difference only in the groups aged 6 to 12 and 13 to 18 years (P < 0.01). CONCLUSION: The prevalence of dental caries, gingival bleeding and calculus in disabled children and adolescents in China is high. Disability type was strongly associated with oral health status. Specially designed oral health education and training are necessary for clinical professionals and caregivers.

Targeting soluble epoxide hydrolase promotes osteogenic-angiogenic coupling via activating SLIT3/HIF-1α signalling pathway.

Type H vessels have recently been identified to modulate osteogenesis. Epoxyeicostrioleic acids (EETs) have an essential contribution to vascular homeostasis. However, whether increased EETs with soluble epoxide hydrolase (sEH) inhibitor TPPU enhance the coupling of angiogenesis and osteogenesis remains largely unknown. The effects of TPPU on cross-talk between co-cultured human umbilical vein endothelial cells (HUVECs) and human dental pulp stem cells (hDPSCs), and on long bone growth and calvarial defect repair in mice were investigated in vitro and in vivo. TPPU enhanced osteogenic differentiation of co-cultured HUVECs and hDPSCs in vitro and increased type H vessels, and long bone growth and bone repair of calvarial defect. Mechanistically, TPPU promoted cell proliferation and angiogenesis, reclined cell apoptosis, and significantly increased CD31hi EMCNhi endothelial cells (ECs) and SLIT3 and HIF-1α expression levels in co-cultured HUVECs and hDPSCs. Knockdown of Slit3 in hDPSCs or Hif-1α in HUVECs impaired the formation of CD31hi EMCNhi ECs and reversed TPPU-induced osteogenesis. We defined a previously unidentified effect of TPPU coupling angiogenesis and osteogenesis. TPPU induced type H vessels by upregulating the expression of hDPSCs-derived SLIT3, which resulted in the activation of ROBO1/YAP1/HIF-1α signalling pathway in ECs. Targeting metabolic pathways of EETs represents a new strategy to couple osteogenesis and angiogenesis, sEH is a promising therapeutic target for bone regeneration and repair.

Aluminum-induced oxidative stress and changes in antioxidant defenses in the roots of rice varieties differing in Al tolerance.

The effects of aluminum (Al) on root elongation, lipid peroxidation, hydrogen peroxide (H(2)O(2)) accumulation, antioxidant levels, antioxidant enzymatic activity, and lignin content in the roots of the Al-tolerant rice variety azucena and the Al-sensitive variety IR64 were investigated. Treatment with Al induced a greater decrease in root elongation and a greater increase in H(2)O(2) and lipid peroxidation as determined by the total thiobarbituric acid-reactive substance (TBARS) level in IR64 than in azucena. Azucena had significantly higher levels of superoxide dismutase, ascorbate peroxidase, glutathione reductase, and glutathione peroxidase GSH POD activity compared with IR64. The concentrations of reduced glutathione (GSH) and ascorbic acid, and the GSH/GSSG ratio (reduced vs. oxidized glutathione) were also higher in azucena than in IR64 in the presence of Al. The addition of 1 mg/L GSH improved root elongation in both varieties and decreased H(2)O(2) production under Al stress. By contrast, treatment with buthionine sulfoximine, a specific inhibitor of GSH synthesis, decreased root elongation in azucena and stimulated H(2)O(2) production in both varieties. Moreover, Al treatment significantly increased the cytoplasmic activity of peroxidase (POD) as well as the levels of POD bound ionically and covalently to cell walls in the Al-sensitive variety. The lignin content was also increased. Treatment with exogenous H(2)O(2) also increased the lignin content and decreased root elongation in IR64. These results suggest that Al induces lignification in the roots of Al-sensitive rice varieties, probably through an increase in H(2)O(2) accumulation.

Status Quo and Advanced Progress in Oral Health Care and Treatment of Children with Autism Spectrum Disorder: A Literature Review.

Autism spectrum disorder (ASD) has become one of the fastest growing diseases in the world, causing a great burden to ASD children's families and society. Children with ASD face more disadvantages relating to their oral health than those without ASD. There is a positive correlation between prevalence of caries lesions and severity of ASD. Poorer oral hygiene, higher detection rates of dental calculus and far more frequent cases of gingivitis occur in children with ASD. Traumatic injuries and various types of malocclusions are more frequent in children with ASD. Poorer oral health care and treatment status are caused by multiple adverse factors. Ways of promoting effective oral health care and treatment include pretreatment counselling; improvement of the individualised treatment environment; routine behaviour guidance techniques (BGTs) including tell-show-do, distraction, role model presentation, voice control, visual education and social stories, encouragement and reinforcement; targeted BGTs including visual education, behaviour modelling, applied behaviour analysis (ABA) and systematic desensitisation; passive BGTs including protective restraint, pharmaceutically administrated sedation and general anaesthesia; oral health education for guardians; and interdisciplinary collaboration and professional dental care/treatment. Dentists, families with children with ASD and schools should cooperate to improve family-centred oral health care and treatment for ASD children not only in China, but also the whole world.

In vivo methods for evaluating human midpalatal suture maturation and ossification: An updated review.

OBJECTIVES: The aim of this study was to perform an updated review of the in vivo methods to evaluate human midpalatal suture maturation and ossification, since this evaluation process remains an unsolved and critical problem in orthodontic treatment. MATERIALS AND METHODS: PubMed, Embase, Cochrane Library, Scopus and Web of Science databases were searched up to November 30, 2021. Literature selection was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA, 2020 Edition) statement and was based on predetermined inclusion criteria. The overall and methodological characteristics of the selected studies were collected. The risk of bias was evaluated mainly through inter- and intra-evaluator agreement outcomes reported in each study. As there was a high heterogeneity among methodological studies, meta-analysis of the included studies was not applicable, and results were analysed descriptively. RESULTS: Nine articles met the inclusion criteria. Maxillary occlusal radiograph and computed tomography (CT), especially cone beam CT (CBCT), were reported. The occlusal radiograph is not adequate for evaluating the status of midpalatal suture maturation, and has been replaced by CBCT. Qualitative and quantitative CBCT evaluation methods provide limited evidence; however, opinions differ regarding the efficacy of these methods. CONCLUSIONS: For midpalatal suture maturation and ossification status evaluation, evidence for the current methods is still limited. Further methodological studies should use image information comprehensively and provide verification evidence on larger samples.

Lignin valorization: Status, challenges and opportunities.

As an abundant aromatic biopolymer, lignin has the potential to produce various chemicals, biofuels of interest through biorefinery activities and is expected to benefit the future circular economy. However, lignin valorization is hindered by a series of constraints such as heterogeneous polymeric nature, intrinsic recalcitrance, strong smell, dark colour, challenges in lignocelluloses fractionation and the presence of high bond dissociation enthalpies in its functional groups etc. Nowadays, industrial lignin is mostly combusted for electricity production and the recycling of inorganic compounds involved in the pulping process. Given the research and development on lignin valorization in recent years, important applications such as lignin-based hydrogels, surfactants, three-dimensional printing materials, electrodes and production of fine chemicals have been systematically reviewed. Finally, this review highlights the main constraints affecting industrial lignin valorization, possible solutions and future perspectives, in the light of its abundance and its potential applications reported in the scientific literature.

Identifying Facial Features and Predicting Patients of Acromegaly Using Three-Dimensional Imaging Techniques and Machine Learning.

Objective: Facial changes are common among nearly all acromegalic patients. As they develop slowly, patients often fail to notice such changes before they become obvious. Consequently, diagnosis and treatment are often delayed. So far, convenient and accurate early detection of this disease is still unavailable. This study is designed to combine the use of 3D imaging and machine learning techniques in facial feature analysis and identification of acromegalic patients, in an effort to ascertain how both techniques performed in terms of applicability and value in the early detection of the disease. Methods: One hundred and twenty-four participants including 62 patients with acromegaly and 62 matched controls were enrolled. Using three-dimensional imaging techniques, 58 facial parameters were measured on each face. A two-way analysis of variance (ANOVA) and a post-hoc t-tests were conducted to examine the variations of these parameters with disease status and gender. Using linear discriminant analysis (LDA), we further distinguished patients from controls, characterized what combinations of the parameters could best predict disease state and their relative contributions. Results: Patients are significantly different from normal subjects in many variables, and facial changes of male patients are more significant than female ones. Both male and female patients present following major changes: the increase of facial length and breadth, the widening and elevation of the nose, the thickening of vermilion and the enlargement of the mandible. Facial variables which strongly related to the pathological states can be used to predict the morbid state with high accuracy (prediction accuracies 92.86% in females, p < 0.0001 and 75% in males, p < 0.001). We have further testified that only a few variables play a vital role in disease prediction and the vital combination of variables vary with gender. Conclusions: Three-dimensional imaging enables comprehensive and accurate quantification of facial characteristics, which makes it a promising technique to investigate facial features of acromegalic patients. In combination with machine learning technique, patients can be accurately identified and predicted by their facial variables. This approach might be beneficial for the early detection of acromegalic patients and timely consultation to improve their outcomes.

A novel silicone derivative of natural osalmid (DCZ0858) induces apoptosis and cell cycle arrest in diffuse large B-cell lymphoma via the JAK2/STAT3 pathway.

Diffuse large B-cell lymphoma (DLBCL) is a highly heterogeneous malignant tumor characterized by diffuse growth. DCZ0858 is a novel small molecule with excellent antitumor effects in DLBCL. This study explored in depth the inhibitory effect of DCZ0858 on DLBCL cell lines. Cell Counting Kit-8 (CCK-8) and plate colony formation assays were used to evaluate cell proliferation levels. Flow cytometry was employed to analyze apoptosis and the cell cycle, and western blotting was used to quantify the expression of cell cycle regulators. The results indicated that DCZ0858 inhibited cell growth in a concentration-dependent and time-dependent manner while inducing no significant toxicity in normal cells. Moreover, DCZ0858 initiated cell apoptosis via both internal and external apoptotic pathways. DCZ0858 also induced cell cycle arrest in the G0/G1 phase, thereby controlling cell proliferation. Further investigation of the molecular mechanism showed that the JAK2/STAT3 pathway was involved in the DCZ0858-mediated antitumor effects and that JAK2 was the key target for DCZ0858 treatment. Knockdown of JAK2 partly weakened the DCZ0858-mediated antitumor effect in DLBCL cells, while JAK2 overexpression strengthened the effect of DCZ0858 in DLBCL cells. Moreover, a similar antitumor effect was observed for DCZ0858 and the JAK2 inhibitor ruxolitinib, and combining the two could significantly enhance cancer-suppressive signaling. Tumor xenograft models showed that DCZ0858 inhibited tumor growth in vivo and had low toxicity in important organs, findings that were consistent with the in vitro data. In summary, DCZ0858 is a promising drug for the treatment of DLBCL.

Immersive virtual reality as analgesia for women during hysterosalpingography: study protocol for a randomized controlled trial.

BACKGROUND: Hysterosalpingography (HSG) is an accepted radiologic diagnostic modality for initial infertility workup, and is generally considered uncomfortable and painful. However, the management of pain related to HSG remains inefficient. As an emerging nonpharmacologic and noninvasive pain control strategy, virtual reality (VR) distraction has been successfully used in areas such as burns, blunt force trauma, hospital-based needle procedures, dental/periodontal procedures, and urological endoscopy patients. This study aims to evaluate the analgesic effect of VR during HSG. METHODS/DESIGN: A single-center, parallel-group, randomized controlled trial will be carried out in the Radiology Department of Yinchuan Women and Children Healthcare Hospital, Yinchuan. A total of 200 participants who are scheduled for HSG will be enrolled in this study. The participants will be randomized (1:1) into two groups: a VR group and a blank control group. The VR group will receive routine care plus immersive VR intervention and the blank control group will receive routine care. Outcomes will be monitored at baseline, immediately after HSG and 15 min after HSG for each group. The primary outcome is the worst pain score during HSG by a visual analog scale (VAS). The secondary outcomes include: affective pain, cognitive pain, and anxiety during the HSG procedure; worst pain within 15 min after HSG; patient satisfaction and acceptance with pain management; physiological parameters; adverse effects; HSG results; and immersion perception score of the VR system (for the VR condition only). DISCUSSION: This study will focus on exploring a simply operated, noninvasive and low-cost analgesia during the HSG procedure. The results of this trial will provide data on the feasibility and safety of VR distraction therapy during HSG. TRIAL REGISTRATION: Chinese Clinical Trial Register, ChiCTR1900021342. Registered on 16 February 2019.

Optimal Matrix Preparation Methods for Matrix-assisted Laser Desorption/ionisation Time-of-flight Mass Spectrometry Profiling of Low Molecular Weight Peptides in Human Saliva and Serum Samples.

OBJECTIVE: To develop standard experimental methods to minimise technical variance in matrix preparation for MALDI-TOF MS (matrix-assisted laser desorption/ionisation time-offlight) profiling. METHODS: Matrix factors in saliva and serum samples of 20 healthy volunteers were examined, assuring their peptide components using seven different matrix type/preparation methods, HCCA(a-cyano-4-hydroxycinnamic acid)/SM(sample/matrix), SA(sinapinic acid)/DD(dried droplet), SA/SM, DHB(2.5-dihydroxyhenz-zoic acid)/DD, DHB/SM, DHAP(2.5-dihydroxyacetophenone)/ DD, DHAP/SM. Number of peaks, S/N(signal to noise) ratio and approximate range of target peaks were set as main selection criteria to find if these spell out any common regularity in results. RESULTS: Different methods perform differently. DHB/DD performed worst in both samples, with no effective peak detected. For saliva sample, the S/N ratios of other six methods were lower. M/z range distributed differently. DHB/SM and DHAP/DD performed best in number of peaks, m/z distributing in 1000 to 2000 account for the vast majority. For serum sample, S/N ratios and m/z range distribution were different in different methods. S/N ratio of SA/DD and SA/SM were higher, number of peaks and m/z distribution were not irreplaceable. S/N ratios of the other four methods were lower. CONCLUSION: DHAP/DD and HCCA/SM performed best in number of peaks, m/z in 5000 - 7000 account for the vast majority in HCCA/SM and 1000 - 2000 in DHAP/DD. Further studies should focus on other characteristics of peptide components detected in different matrix methods to increase evidence when selecting matrix type/preparation methods.

A Neutralized Noncharged Polyethylenimine-Based System for Efficient Delivery of siRNA into Heart without Toxicity.

Cationic polymers constitute an important class of materials in development of delivery vehicles for nucleic acid-based therapeutics. Among them, polyethylenimine (PEI) has been a classical cationic carrier intensively studied for therapeutic delivery of DNA, RNA, and short RNA molecules to treat diseases. However, the development of PEI for in vivo applications has been hampered by the inherent problems associated with the material, particularly its cytotoxicity and the instability of the nucleic acid complexation systems formed via electrostatic interactions. Here, we demonstrate a strategy to modify PEI polymers via hydrazidation to create neutralized, stable, and multifunctional system for delivering siRNA molecules. Through substitution of the primary amino groups of PEI with neutral hydrazide groups, cross-linked nanoparticles with surface decorated with a model targeting ligands were generated. The neutral cross-linked siRNA nanoparticles not only showed favorable biocompatibility and cell internalization efficiency in vitro but also allowed for significant tissue uptake and gene silencing efficiency in zebrafish heart in vivo. Our study suggests transformation of conventional branched PEI into a neutral polymer that can lead to a new category of nonviral carriers, and the resulting functional delivery systems may be further explored for development of siRNA therapeutics for treating cardiovascular disease/injury.