Hydroxytryptophan biosynthesis by a family of heme-dependent enzymes in bacteria.

Hydroxytryptophan serves as a chemical precursor to a variety of bioactive specialized metabolites, including the human neurotransmitter serotonin and the hormone melatonin. Although the human and animal routes to hydroxytryptophan have been known for decades, how bacteria catalyze tryptophan indole hydroxylation remains a mystery. Here we report a class of tryptophan hydroxylases that are involved in various bacterial metabolic pathways. These enzymes utilize a histidine-ligated heme cofactor and molecular oxygen or hydrogen peroxide to catalyze regioselective hydroxylation on the tryptophan indole moiety, which is mechanistically distinct from their animal counterparts from the nonheme iron enzyme family. Through genome mining, we also identify members that can hydroxylate the tryptophan indole ring at alternative positions. Our results not only reveal a conserved way to synthesize hydroxytryptophans in bacteria but also provide a valuable enzyme toolbox for biocatalysis. As proof of concept, we assemble a highly efficient pathway for melatonin in a bacterial host.

Discovery of a Bacterial Hydrazine Transferase That Constructs the N-Aminolactam Pharmacophore in Albofungin Biosynthesis.

Structural motifs containing nitrogen-nitrogen (N-N) bonds are prevalent in a large number of clinical drugs and bioactive natural products. Hydrazine (N2H4) serves as a widely utilized building block for the preparation of these N-N-containing molecules in organic synthesis. Despite its common use in chemical processes, no enzyme has been identified to catalyze the incorporation of free hydrazine in natural product biosynthesis. Here, we report that a hydrazine transferase catalyzes the condensation of N2H4 and an aromatic polyketide pathway intermediate, leading to the formation of a rare N-aminolactam pharmacophore in the biosynthesis of broad-spectrum antibiotic albofungin. These results expand the current knowledge on the biosynthetic mechanism for natural products with N-N units and should facilitate future development of biocatalysts for the production of N-N-containing chemicals.

All-Natural Injectable Antibacterial Hydrogel Enabled by Chitosan and Borneol.

Hydrogels with intrinsic antimicrobial capabilities based on natural strategies have been studied as a hot topic in biomedicine. Nevertheless, it is highly challenging to thoroughly develop a bacteriostatic natural hydrogel. Borneol as a traditional Chinese medicine possesses a unique broad-spectrum antibacterial activity under a membrane-breaking mechanism. In this study, a range of fully natural antibacterial hydrogels are designed and synthesized via the Schiff base cross-linking of carboxymethyl chitosan and dialdehyde dextran grafted natural borneol. The borneol with three configurations is hydrophilically modified onto dextran to boost its antibacterial activity. Also, the synergism of hydrophilic-modified borneol groups and positively charged ammonium ions of carboxymethyl chitosan make the hydrogels totally constrict the E. coli and S. aureus growth during 24 h. Furthermore, the hydrogels exhibit good in vitro cytocompatibility through cytotoxicity, protein adhesion, and hemolytic tests. In view of the injectability, the hydrogels can be delivered to the target site through a minimally invasive route. In short, this work offers a potential tactic to develop antibacterial hydrogels for the treatment of topical wound infections.

Regulatory T cells subgroups in the tumor microenvironment cannot be overlooked: Their involvement in prognosis and treatment strategy in melanoma.

BACKGROUND: Melanoma, the most lethal form of skin cancer, presents substantial challenges despite effective surgical interventions for in situ lesions. Regulatory T cells (Tregs) wield a pivotal immunomodulatory influence within the tumor microenvironment, yet their impact on melanoma prognosis and direct molecular interactions with melanoma cells remain elusive. This investigation employs single-cell analysis to unveil the intricate nature of Tregs in human melanoma. METHODS: Single-cell RNA and bulk sequencing data, alongside clinical information, were obtained from public repositories. Initially, GO and GSEA analyses were employed to delineate functional disparities among distinct cell subsets. Pseudotime and cell-cell interconnection analyses were conducted, followed by an endeavor to construct a prognostic model grounded in Treg-associated risk scores. This model's efficacy was demonstrated via PCA and K-M analyses, with multivariate Cox regression affirming its independent prognostic value in melanoma patients. Furthermore, immune infiltration analysis, immune checkpoint gene expression scrutiny, and drug sensitivity assessments were performed to ascertain the clinical relevance of this prognostic model. RESULTS: Following batch effect correction, 80 025 cells partitioned into 31 clusters, encompassing B cells, plasma cells, endothelial cells, fibroblasts, melanoma cells, monocytes, macrophages, and T_NK cells. Within these, 4240 CD4+ T cells were subclassified into seven distinct types. Functional analysis underscored the immunomodulatory function of Tregs within the melanoma tumor microenvironment, elucidating disparities among Treg subpopulations. Notably, the ITGB2 signaling pathway emerged as a plausible molecular nexus linking Tregs to melanoma cells. Our prognostic signature exhibited robust predictive capacities for melanoma prognosis and potential implications in evaluating immunotherapy response. CONCLUSION: Tregs exert a critical role in immune suppression within the melanoma tumor microenvironment, revealing a potential molecular-level association with melanoma cells. Our innovative Treg-centered signature introduces a promising prognostic marker for melanoma, holding potential for future clinical prognostic assessments.

Conserved Enzymatic Cascade for Bacterial Azoxy Biosynthesis.

Azoxy compounds exhibit a wide array of biological activities and possess distinctive chemical properties. Although there has been considerable interest in the biosynthetic mechanisms of azoxy metabolites, the enzymatic basis responsible for azoxy bond formation has remained largely enigmatic. In this study, we unveil the enzyme cascade that constructs the azoxy bond in valanimycin biosynthesis. Our research demonstrates that a pair of metalloenzymes, comprising a membrane-bound hydrazine synthase and a nonheme diiron azoxy synthase, collaborate to convert an unstable pathway intermediate to an azoxy product through a hydrazine-azo-azoxy pathway. Additionally, by characterizing homologues of this enzyme pair from other azoxy metabolite pathways, we propose that this two-enzyme cascade could represent a conserved enzymatic strategy for azoxy bond formation in bacteria. These findings provide significant mechanistic insights into biological N-N bond formation and should facilitate the targeted isolation of bioactive azoxy compounds through genome mining.

Environmental DNA metabarcoding reveals the impacts of anthropogenic pollution on multitrophic aquatic communities across an urban river of western China.

Anthropogenic activities are intensively affecting the structure and function of biological communities in river ecosystems. The effects of anthropogenic pollution on single-trophic community have been widely explored, but their effects on the structures and co-occurrence patterns of multitrophic communities remain largely unknown. In this study, we collected 13 water samples from the Neijiang River in Chengdu City of China, and identified totally 2352 bacterial, 207 algal, 204 macroinvertebrate, and 33 fish species based on the eDNA metabarcoding to systematically investigate the responses of multitrophic communities to environmental stressors. We observed significant variations in bacterial, algal, and macroinvertebrate community structures (except fish) with the pollution levels in the river. Network analyses indicated a more intensive interspecific co-occurrence pattern at high pollution level. Although taxonomic diversity of the multitrophic communities varied insignificantly, phylogenetic diversities of fish and algae showed significantly positive and negative associations with the pollution levels, respectively. We demonstrated the primary role of environmental filtering in driving the structures of bacteria, algae, and macroinvertebrates, while the fish was more controlled by dispersal limitation. Nitrogen was identified as the most important factor impacting the multitrophic community, where bacterial composition was mostly associated with NO3--N, algal spatial differentiation with TN, and macroinvertebrate and fish with NH4+-N. Further partial least-squares path model confirmed more important effect of environmental variables on the relative abundance of bacteria and algae, while macroinvertebrate and fish communities were directly driven by the algae-mediated pathway in the food web. Our study highlighted the necessity of integrated consideration of multitrophic biodiversity for riverine pollution management, and emphasized the importance of controlling nitrogen inputs targeting a healthy ecosystem.

Experimental Study of Skin Contraction Induced by Bipolar Radiofrequency.

Background: Facial skin relaxation has become an important part in solving the problem of facial rejuvenation. Minimally invasive or noninvasive skin-tightening procedures have become a trend for facial rejuvenation. Bipolar radiofrequency (RF) is a new option for treating skin relaxation and is more effective than noninvasive surgery without surgical incision. Objective: To explore the effect of different bipolar RF powers on the area of the original box, changes of skin and subcutaneous tissue thickness and numbers of fibroblasts in rabbits. Design: The research team performed an animal study. Setting: This study took place in Affiliated Hospital of Nanjing University of Chinese Medicine. Participants: Eighteen common-grade adult New Zealand rabbits (female, 2.5-3.0 kg). Methods: Bipolar radiofrequency therapy was given to a girl rabbit on the left side of the treatment area. Standard HE and Masson staining were performed to assess the pathological changes, area of the original box and the number of fibroblasts in skin and subcutaneous tissues. Outcome Measures: (1) The area of the original box, changes of skin and subcutaneous tissue thickness, and numbers of fibroblasts under different bipolar RF temperatures or under different bipolar RF powers immediately after surgery, 1 month after surgery and 3 months after surgery were observed. (2) Standard HE and Masson staining results. Results: Under the condition of certain instrument power, at 36de 38d and 40nd the area of the original box shrank to different degrees immediately after surgery (16.54±0.37, 17.78±0.03, 17.19±0.01), 1 month after surgery (16.59±0.31, 17.82±0.01, 18.34±0.30) and 3 months after surgery (16.89±0.12, 18.16±0.14, 19.23±0.32) compared with that before surgery (P < .05). Under specific temperature conditions, at 16 W, 18 W, 20 W, and 22 W, the area of the original box shrank to different degrees immediately after surgery (16.40±0.49, 15.55±0.57, 17.54±0.12, 16.19±0.27), 1 month after surgery (16.88±0.12, 17.46±0.02, 18.05±0.35, 19.41±0.08) and 3 months after surgery (19.09±1.01, 18.30±0.69, 20.00±0.29, 21.20±0.90) compared with that before surgery (P < .05). When the power was fixed, the thickness of skin and subcutaneous tissue decreased immediately after surgery (6.7, 6.8, 7), 1 month after surgery (6, 6.1, 6.3) and 3 months after surgery (6.4, 6.5, 6.2) at different temperatures (P < .05). When the temperature was fixed, the thickness of skin and subcutaneous tissue decreased immediately after surgery (6.1, 6.08, 6.03), 1 month after surgery (6.2, 6.15, 6.13), and 3 months after surgery (6.2, 6.23, 6.03) under different powers (P < .05). Under the condition of certain instrument power, at 36de 38d and 40n, the number of fibroblasts increased to different degrees immediately after surgery (26.54±2.37, 30.78±3.03, 37.19±4.01), 1 month after surgery (28.59±2.31, 34.82±3.01, 40.34±4.30), and 3 months after surgery (30.89±0.12, 38.16±0.14, 42.23±0.32) compared with that before surgery, and all were statistically significant (P < .05). Under specific temperature conditions, at 16 W, 18 W, 20 W, and 22 W, the number of fibroblasts increased to different degrees immediately after surgery (28.29±2.49, 30.97±3.57, 38.74±3.12, 45.68±4.27), 1 month after surgery (30.88±3.12, 32.46±4.02, 41.05±0.35, 50.41±0.08), and 3 months after surgery (29.99±2.01, 33.30±2.69, 39.00±3.29, 23.20±2.90) compared with that before surgery, and all were statistically significant (P < .05). Conclusions: Our study clarifies that bipolar RF can decrease the skin and subcutaneous tissue thickness and increase the numbers of fibroblasts at the temperature of 36°C, 38°C, and 40°C and frequency of 16-22 W, which has a therapeutical effect on skin contraction. Our study might effectively improve the skin slack of patients, and the postoperative maintenance rate is high, and will not cause obvious complications. This study may provide a theoretical direction for clinicians to tighten the skin of patients using bipolar RF.

Advances in Robot-Assisted Surgery for Facial Bone Contouring Surgery.

Since our team reported the application of robot-assisted surgery in facial contouring surgery in 2020, further clinical trials with large samples have been conducted. This paper will report the interim results of a single-center, large-sample randomized controlled trial of the first robot developed by our team for facial contouring surgery. Meanwhile, this research field will be systematically reviewed and prospected.

Gelatin-Reinforced Zwitterionic Organohydrogel with Tough, Self-Adhesive, Long-Term Moisturizing and Antifreezing Properties for Wearable Electronics.

Strong mechanical performance, appropriate adhesion capacity, and excellent biocompatibility of conductive hydrogel-based sensors are of great significance for their application. However, conventional conductive hydrogels usually exhibit insufficient mechanical strength and adhesion. In addition, they will lose flexibility and conductivity under subzero temperature and a dry environment owing to inevitable freezing and evaporation of water. In this study, a tough, flexible, self-adhesive, long-term moisturizing, and antifreezing organohydrogel was prepared, which was composed of gelatin, zwitterionic poly [2-(methacryloyloxy) ethyl] dimethyl-(3-sulfopropyl) (PSBMA), MXene nanosheets, and glycerol. Natural gelatin was incorporated to enhance mechanical performance via the entanglement of a physical cross-linked network and a PSBMA network, which was also used as a stabilizer to disperse MXene into the organohydrogel. Zwitterionic PSBMA endowed the organohydrogel with good adhesion and self-healing properties. Long-term moisturizing properties and antifreeze tolerance could be achieved owing to the synergistic water retention capacity of PSBMA and glycerol. The resulting PSBMA-gelatin-MXene-glycerol (PGMG) organohydrogel exhibited high mechanical fracture strength (0.65 MPa) and stretchability (over 1000%), excellent toughness (3.87 MJ/m3), strong and repeated adhesion to diverse substrates (e.g., paper, glass, silicon rubber, iron, and pig skin), good fatigue resistance (under the cyclic stretching-releasing process), and rapid recovery capacity. Moreover, the PGMG organohydrogel showed good stability under -40 °C. The sensor based on PGMG organohydrogel could tightly attach to the human skin and real-time-monitor the motions of joints (e.g., bending of the finger, wrist, elbow, and knee) and the change in mood such as smiling and frowning. Therefore, PGMG organohydrogels have a huge potential for wearable sensors under room temperature or extreme environments.

Absolute Ethanol Embolization of Lip Arteriovenous Malformations: Observational Results from 10 Years of Experience.

PURPOSE: To evaluate the safety and efficacy of ethanol embolization of lip arteriovenous malformations (AVMs). MATERIALS AND METHODS: Seventy-six patients with lip AVMs were treated with 173 ethanol embolization procedures. Lip AVMs were treated with direct puncture alone in 21 patients (35 procedures, 20.2%), transarterial embolization alone in 13 patients (18 procedures, 10.4 %), and a combination of both in 60 patients (120 procedures, 69.3%). Adjunctive surgical resection was performed after embolization for cosmetic purposes based on the patient's request, including patient preference, functional impairment, and skin necrosis. The mean duration of follow-up was 30.9 months ± 27.6. The follow-up included clinic visits and telephonic questionnaires to evaluate the clinical signs and symptoms of AVMs as well as quality of life measures. RESULTS: Of 76 patients, 51 showed 100% devascularization of AVMs, as determined using arteriography, followed by 23 with 76%-99% devascularization and 2 with 50%-75% devascularization. Of the 76 patients, 40 achieved complete symptom relief and 25 achieved major improvements in cosmetic deformity after embolization. Additionally, 54 patients achieved satisfactory function and aesthetic improvement with ethanol embolotherapy alone, whereas 22 achieved similar outcomes with a combination of ethanol embolotherapy and surgical intervention. Thirty-three adverse events (including 1 major) were documented. CONCLUSIONS: Ethanol embolization of lip AVMs, as a mainstay, is efficacious in managing these lesions, with acceptable complications. Surgical resection after embolization may improve function and cosmesis in a subset of patients.

Using the hybrid EMD-BPNN model to predict the incidence of HIV in Dalian, Liaoning Province, China, 2004-2018.

BACKGROUND: Acquired immunodeficiency syndrome (AIDS) is a malignant infectious disease with high mortality caused by HIV (human immunodeficiency virus, and up to now there are no curable drugs or effective vaccines. In order to understand AIDS's development trend, we establish hybrid EMD-BPNN (empirical modal decomposition and Back-propagation artificial neural network model) model to forecast new HIV infection in Dalian and to evaluate model's performance. METHODS: The monthly HIV data series are decomposed by EMD method, and then all decomposition results are used as training and testing data to establish BPNN model, namely BPNN was fitted to each IMF (intrinsic mode function) and residue separately, and the predicted value is the sum of the predicted values from the models. Meanwhile, using yearly HIV data to established ARIMA and using monthly HIV data to established BPNN, and SARIMA (seasonal autoregressive integrated moving average) model to compare the predictive ability with EMD-BPNN model. RESULTS: From 2004 to 2017, 3310 cases of HIV were reported in Dalian, including 101 fatal cases. The monthly HIV data series are decomposed into four relatively stable IMFs and one residue item by EMD, and the residue item showed that the incidence of HIV increases firstly after declining. The mean absolute percentage error value for the EMD-BPNN, BPNN, SARIMA (1,1,2) (0,1,1)12 in 2018 is 7.80%, 10.79%, 9.48% respectively, and the mean absolute percentage error value for the ARIMA (3,1,0) model in 2017 and 2018 is 8.91%. CONCLUSIONS: The EMD-BPNN model was effective and reliable in predicting the incidence of HIV for annual incidence, and the results could furnish a scientific reference for policy makers and health agencies in Dalian.

Prognostic Value of Different Computed Tomography Scoring Systems in Patients With Severe Traumatic Brain Injury Undergoing Decompressive Craniectomy.

OBJECTIVE: In this study, we investigate the preoperative and postoperative computed tomography (CT) scores in severe traumatic brain injury (TBI) patients undergoing decompressive craniectomy (DC) and compare their predictive accuracy. METHODS: Univariate and multivariate logistic regression analyses were used to determine the relationship between CT score (preoperative and postoperative) and mortality at 30 days after injury. The discriminatory power of preoperative and postoperative CT score was assessed by the area under the receiver operating characteristic curve (AUC). RESULTS: Multivariate logistic regression analysis adjusted for the established predictors of TBI outcomes showed that preoperative Rotterdam CT score (odds ratio [OR], 3.60; 95% confidence interval [CI], 1.13-11.50; P = 0.030), postoperative Rotterdam CT score (OR, 4.17; 95% CI, 1.63-10.66; P = 0.003), preoperative Stockholm CT score (OR, 3.41; 95% CI, 1.42-8.18; P = 0.006), postoperative Stockholm CT score (OR, 4.50; 95% CI, 1.60-12.64; P = 0.004), preoperative Helsinki CT score (OR, 1.44; 95% CI, 1.03-2.02; P = 0.031), and postoperative Helsinki CT score (OR, 2.55; 95% CI, 1.32-4.95; P = 0.005) were significantly associated with mortality. The performance of the postoperative Rotterdam CT score was superior to the preoperative Rotterdam CT score (AUC, 0.82-0.97 vs 0.71-0.91). The postoperative Stockholm CT score was superior to the preoperative Stockholm CT score (AUC, 0.76-0.94 vs 0.72-0.92). The postoperative Helsinki CT score was superior to the preoperative Helsinki CT score (AUC, 0.88-0.99 vs 0.65-0.87). CONCLUSIONS: In conclusion, assessing the CT score before and after DC may be more precise and efficient for predicting early mortality in severe TBI patients who undergo DC.

Initial CT-based radiomics nomogram for predicting in-hospital mortality in patients with traumatic brain injury: a multicenter development and validation study.

OBJECTIVE: To develop and validate a radiomic prediction model using initial noncontrast computed tomography (CT) at admission to predict in-hospital mortality in patients with traumatic brain injury (TBI). METHODS: A total of 379 TBI patients from three cohorts were categorized into training, internal validation, and external validation sets. After filtering the unstable features with the minimum redundancy maximum relevance approach, the CT-based radiomics signature was selected by using the least absolute shrinkage and selection operator (LASSO) approach. A personalized predictive nomogram incorporating the radiomic signature and clinical features was developed using a multivariate logistic model to predict in-hospital mortality in patients with TBI. The calibration, discrimination, and clinical usefulness of the radiomics signature and nomogram were evaluated. RESULTS: The radiomic signature consisting of 12 features had areas under the curve (AUCs) of 0.734, 0.716, and 0.706 in the prediction of in-hospital mortality in the internal and two external validation cohorts. The personalized predictive nomogram integrating the radiomic and clinical features demonstrated significant calibration and discrimination with AUCs of 0.843, 0.811, and 0.834 in the internal and two external validation cohorts. Based on decision curve analysis (DCA), both the radiomic features and nomogram were found to be clinically significant and useful. CONCLUSION: This predictive nomogram incorporating the CT-based radiomic signature and clinical features had maximum accuracy and played an optimized role in the early prediction of in-hospital mortality. The results of this study provide vital insights for the early warning of death in TBI patients.

Zwitterionic Hydrogel with High Transparency, Ultrastretchability, and Remarkable Freezing Resistance for Wearable Strain Sensors.

Multifunctional hydrogel with outstanding conductivity and mechanical flexibility has received enormous attention as wearable electronic devices. However, fabricating transparent, ultrastretchable, and biocompatible hydrogel with low-temperature stability still remains a tremendous challenge. In this study, an ultrastretchable, highly transparent, and antifreezing zwitterionic-based electronic sensor is developed by introducing zwitterionic proline (ZP) into gellan gum/polyacrylamide (GG/PAAm) double network (DN) hydrogel. The existence of ZP endows the hydrogel with remarkable frost resistance. The toughness and transparency of zwitterionic Ca-GG/PAAm-ZP DN hydrogel can be maintained down to -40 °C. Also, the zwitterionic hydrogel shows good biocompatibility and protein adsorption resistance. The zwitterionic Ca-GG/PAAm-ZP DN hydrogel-based strain sensor can accurately monitor human motions (such as speaking and various joint bendings) under a broad temperature range from -40 to 25 °C. The zwitterionic Ca-GG/PAAm-ZP DN hydrogel-based strain sensor will be of immense value in the field of wearable electronic devices, especially for extreme environment applications.

Study of the migration behaviours of imidazoline corrosion inhibitor in concrete using UV-vis spectrophotometry.

In this study, UV-visible light spectrophotometry was used for the first time to examine the migration behaviours of cationic and nonionic imidazoline corrosion inhibitors in concrete. Imidazoline can react with bromocresol purple resulting in a reduction in absorbance, which can be used to calculate quantitatively imidazoline concentration. The results showed that the migration rate of nonionic imidazoline was faster than that of cationic imidazoline with or without the presence of an electric field, possibly because of the better water solubility of nonionic imidazoline. The electric field could significantly accelerate the migration rate of the cationic and nonionic imidazoline. However, the penetration performance of nonionic imidazoline was much improved compared with that of cationic imidazoline in concrete. From X-ray photoelectron spectroscopy analysis, the N element could be detected on steel, verifying the migration behaviour of the nonionic imidazoline. This nonionic imidazoline could markedly retard steel corrosion according to potentiodynamic polarization.

Versatile Roles of Aquaporins in Plant Growth and Development.

Aquaporins (AQPs) are universal membrane integrated water channel proteins that selectively and reversibly facilitate the movement of water, gases, metalloids, and other small neutral solutes across cellular membranes in living organisms. Compared with other organisms, plants have the largest number of AQP members with diverse characteristics, subcellular localizations and substrate permeabilities. AQPs play important roles in plant water relations, cell turgor pressure maintenance, the hydraulic regulation of roots and leaves, and in leaf transpiration, root water uptake, and plant responses to multiple biotic and abiotic stresses. They are also required for plant growth and development. In this review, we comprehensively summarize the expression and roles of diverse AQPs in the growth and development of various vegetative and reproductive organs in plants. The functions of AQPs in the intracellular translocation of hydrogen peroxide are also discussed.

[Comprehensive Evaluation of Biological Activity in Different Passage Populations of Mesenchymal Stem Cells Derived from Bone Marrow in Ovariectomy Osteoporotic Rats].

This study aimed to comprehensively evaluate the biological activity in different passage populations of mesenchymal stem cells(BMSCs)derived from bone marrow in ovariectomy osteoporotic rats(named OVX-rBMSCs),providing experimental basis for new osteoporotic drug development and research.OVX-rBMSCs were isolated and cultured in vitro by the whole bone marrow adherent screening method.The morphological observation,cell surface markers(CD29,CD45,CD90)detection,cell proliferation,induced differentiation experimental detection were performed to evaluate the biological activity of Passage 1,2,3,4populations(P1,P2,P3,P4)OVX-rBMSCs.The results showed that whole bone marrow adherent culture method isolated and differentially subcultured OVXrBMSCs.The morphology of P4 OVX-rBMSCs was identical fibroblast-like and had the characteristics of ultrastructure of stem cells.The CD29 positive cells rate,CD90 positive cells rate,cell proliferation index,and the osteogenic,adipogenic,chondrogenic differentiation capacities of P4 OVX-rBMSCs were significantly better than those of other populations(P<0.05).OVX-rBMSCs purity and biological activity were gradually optimized with the passaged,and among them P4 cells were superior to all the other populations.Based on these results,we report that the P4OVX-rBMSCs model developed in this study can be used to develop a new and effective medical method for osteoporotic drug screening.

Developing targeted therapies for neuroblastoma by dissecting the effects of metabolic reprogramming on tumor microenvironments and progression.

Rationale: Synergic reprogramming of metabolic dominates neuroblastoma (NB) progression. It is of great clinical implications to develop an individualized risk prognostication approach with stratification-guided therapeutic options for NB based on elucidating molecular mechanisms of metabolic reprogramming. Methods: With a machine learning-based multi-step program, the synergic mechanisms of metabolic reprogramming-driven malignant progression of NB were elucidated at single-cell and metabolite flux dimensions. Subsequently, a promising metabolic reprogramming-associated prognostic signature (MPS) and individualized therapeutic approaches based on MPS-stratification were developed and further validated independently using pre-clinical models. Results: MPS-identified MPS-I NB showed significantly higher activity of metabolic reprogramming than MPS-II counterparts. MPS demonstrated improved accuracy compared to current clinical characteristics [AUC: 0.915 vs. 0.657 (MYCN), 0.713 (INSS-stage), and 0.808 (INRG-stratification)] in predicting prognosis. AZD7762 and etoposide were identified as potent therapeutics against MPS-I and II NB, respectively. Subsequent biological tests revealed AZD7762 substantially inhibited growth, migration, and invasion of MPS-I NB cells, more effectively than that of MPS-II cells. Conversely, etoposide had better therapeutic effects on MPS-II NB cells. More encouragingly, AZD7762 and etoposide significantly inhibited in-vivo subcutaneous tumorigenesis, proliferation, and pulmonary metastasis in MPS-I and MPS-II samples, respectively; thereby prolonging survival of tumor-bearing mice. Mechanistically, AZD7762 and etoposide-induced apoptosis of the MPS-I and MPS-II cells, respectively, through mitochondria-dependent pathways; and MPS-I NB resisted etoposide-induced apoptosis by addiction of glutamate metabolism and acetyl coenzyme A. MPS-I NB progression was fueled by multiple metabolic reprogramming-driven factors including multidrug resistance, immunosuppressive and tumor-promoting inflammatory microenvironments. Immunologically, MPS-I NB suppressed immune cells via MIF and THBS signaling pathways. Metabolically, the malignant proliferation of MPS-I NB cells was remarkably supported by reprogrammed glutamate metabolism, tricarboxylic acid cycle, urea cycle, etc. Furthermore, MPS-I NB cells manifested a distinct tumor-promoting developmental lineage and self-communication patterns, as evidenced by enhanced oncogenic signaling pathways activated with development and self-communications. Conclusions: This study provides deep insights into the molecular mechanisms underlying metabolic reprogramming-mediated malignant progression of NB. It also sheds light on developing targeted medications guided by the novel precise risk prognostication approaches, which could contribute to a significantly improved therapeutic strategy for NB.

NORAD accelerates skin wound healing through extracellular vesicle transfer from hypoxic adipose derived stem cells: miR-524-5p pathway and Pumilio protein mechanism.

Skin wound healing is a multifaceted biological process that encompasses a variety of cell types and intricate signaling pathways. Recent research has uncovered that exosomes derived from adipose stem cells, commonly referred to as ADSC exosomes, play a crucial role in facilitating the healing process. Moreover, it has been demonstrated that an anoxic, or low-oxygen, environment significantly enhances the effectiveness of these exosomes in promoting skin repair. The primary objective of this study was to investigate the underlying mechanisms through which ADSC exosomes contribute to Skin wound healing, particularly by regulating the long non-coding RNA known as NORAD under hypoxic conditions. A significant focus of our research was to examine the interplay between the microRNA miR-524-5p and the Pumilio protein, as we aimed to understand how these molecular interactions might influence the overall healing process. In this study, ADSC exosomes were extracted by simulating hypoxia in vitro and their effects on the proliferation and migration of skin fibroblasts (FB) were evaluated. The expression levels of NORAD, miR-524-5p and Pumilio were analyzed by fluorescence quantitative PCR. Pumilio protein was silenced by siRNA technique to evaluate its role in ADSC exosome-mediated wound healing. The experimental results showed that under hypoxia conditions, NORAD levels in ADSC exosomes increased significantly and could effectively regulate the expression of miR-524-5p. After Pumilio protein silencing, the proliferation and migration ability of fibroblasts were significantly reduced, indicating that Pumilio protein played a role in the process of wound healing. By inhibiting miR-524-5p, the expression of Pumilio protein was restored, further confirming its regulatory mechanism.

Intelligent electromagnetic navigation system for robot-assisted intraoral osteotomy in mandibular tumor resection: a model experiment.

Background: Mandibular tumor surgery necessitates precise osteotomies based on tumor boundaries; however, conventional osteotomies often lack accuracy in predicting osteotomy positions and planes, potentially leading to excessive resection of normal bone tissues or residual tumors, thus compromising postoperative quality of life and clinical outcomes. Robotic-assisted surgery (RAS) augmented with artificial intelligence (AI) offers precise localization capabilities, aiding surgeons in achieving accurate osteotomy positioning. This study aimed to evaluate the feasibility and accuracy of a robotic magnetic navigation system for positioning and osteotomy in an intraoral surgical trial of a mandibular tumor model. Methods: Patient computed tomography (CT) imaging data of mandibular chin and body tumors were utilized to create 3D printed models, serving as study subjects for mandibular tumor resection. Ten pairs of models were printed for the experimental and control groups. The experimental group (EG) underwent osteotomy using a robot-assisted surgical navigation system, performing osteotomy under robotic navigation following alignment based on preoperative design. The control group (CG) underwent traditional surgery, estimating osteotomy position empirically according to preoperative design. Postoperative CT scans were conducted on both models, and actual postoperative results were compared to preoperative design. Osteotomy accuracy was evaluated by positional and angular errors between preoperatively designed and actual osteotomy planes. Results: For ten randomly selected spots on the left and right sides, respectively, the EG group had mean distance errors of 0.338 mm and 0.941 mm. These values were obtained from the EG group. In the EG group, on the left side, the mean angular errors were 14.741 degrees, while on the right side, they were 13.021 degrees. For the 10 randomly selected spots on the left and right sides, respectively, the CG had mean distance errors of 1.776 mm and 2.320 mm. This is in contrast to the results obtained by the EG. It was determined that the left side had a mean angle error of 16.841 degrees, while the right side had an error of 18.416 degrees in the CG group. The above results indicated significantly lower point errors of bilateral osteotomy planes in the experimental group compared to the control group. Conclusion: This study demonstrates the feasibility of electromagnetic navigation robot-assisted intraoral osteotomy for mandibular tumors and suggests that this approach can enhance the precision of clinical surgery.