Tensile Stress-Activated and Exosome-Transferred YAP/TAZ-Notch Circuit Specifies Type H Endothelial Cell for Segmental Bone Regeneration.

The Ilizarov technique has been continuously innovated to utilize tensile stress (TS) for inducing a bone development-like regenerative process, aiming to achieve skeletal elongation and reconstruction. However, it remains uncertain whether this distraction osteogenesis (DO) process induced by TS involves the pivotal coupling of angiogenesis and osteogenesis mediated by type H endothelial cells (THECs). In this study, it is demonstrated that the Ilizarov technique induces the formation of a metaphysis-like architecture composed of THECs, leading to segmental bone regeneration during the DO process. Mechanistically, cell-matrix interactions-mediated activation of yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ) transcriptionally upregulates the expression of Notch1 and Delta-like ligand 4, which act as direct positive regulators of THECs phenotype, in bone marrow endothelial cells (BMECs) upon TS stimulation. Simultaneously, the Notch intracellular domain enhances YAP/TAZ activity by transcriptionally upregulating YAP expression and stabilizing TAZ protein, thus establishing the YAP/TAZ-Notch circuit. Additionally, TS-stimulated BMECs secrete exosomes enriched with vital molecules in this positive feedback pathway, which can be utilized to promote segmental bone defect healing, mimicking the therapeutic effects of Ilizarov technique. The findings advance the understanding of TS-induced segmental bone regeneration and establish the foundation for innovative biological therapeutic strategies aimed at activating THECs.

Enhanced Bone Regeneration through Regulation of Mechanoresponsive FAK-ERK1/2 Signaling by ZINC40099027 during Distraction Osteogenesis.

Background: Focal adhesion kinase (FAK) is activated by mechanical stimulation and plays a vital role in distraction osteogenesis (DO), a well-established but lengthy procedure for repairing large bone defects. Both angiogenesis and osteogenesis contribute to bone regeneration during DO. However, the effects of ZINC40099027 (ZN27), a potent FAK activator, on angiogenesis, osteogenesis, and bone regeneration in DO remain unknown. Methods: The angiogenic potential of human umbilical vein endothelial cells (HUVECs) was evaluated using transwell migration and tube formation assays. The osteogenic activity of bone marrow mesenchymal stem cells (BMSCs) was assessed using alkaline phosphatase (ALP) and alizarin red s (ARS) staining. Additionally, quantitative real-time polymerase chain reaction (qRT-PCR), western blot, and immunofluorescence staining were used to assay angiogenic markers, osteogenic markers, and FAK-extracellular signal-regulated kinase 1/2 (ERK1/2) signaling. In vivo, a rat tibia DO model was established to verify the effects of ZN27 on neovascularization and bone regeneration using radiological and histological analyses. Results: ZN27 promoted the migration and angiogenesis of HUVECs. Additionally, ZN27 facilitated the osteogenic differentiation of BMSCs, as revealed by increased ALP activity, calcium deposition, and expression of osteogenesis-specific markers. The ERK1/2-specific inhibitor PD98059 significantly hindered the effects of ZN27, suggesting the participation of FAK-ERK1/2 signaling in ZN27-enhanced angiogenesis and osteogenesis. As indicated by improved radiological and histological features, ZN27 induced active angiogenesis within the distraction area and accelerated bone regeneration in a rat DO model. Conclusion: Our results show that ZN27 targets FAK-ERK1/2 signaling to stimulate both angiogenesis and osteogenesis, and ZN27 accelerates bone regeneration in DO, suggesting the therapeutic potential of ZN27 for repairing large bone defects in the mechanobiological environment during DO.

Study on maintenance of eyeball morphology by foldable capsular vitreous body in severe ocular trauma.

OBJECTIVES: To explore the feasibility and safety of using a foldable capsular vitreous body (FCVB) in managing severe ocular trauma and silicone oil-dependent eyes. METHODOLOGY: This is a retrospective study of 61 ocular trauma patients (61 eyes) who presented to the Department of Eye Emergency, Hebei Eye Hospital from May 1, 2018, to May 31, 2019, including 51 male patients (51 eyes) and 10 female patients (10 eyes) with an average age of 44.98 ± 14.60 years old. The oldest patient was 75 years old, and the youngest was 8 years old. These cases represented 51 eyes with severe eyeball rupture and 10 eyes with severe, complicated ocular trauma, which became silicone oil-dependent after the operation. These patients received FCVB implants, and data regarding their visual acuity, intraocular pressure, changes in eye axis, cornea, retina, and FCVB state were recorded after the operation. RESULTS: In all patients, the FCVB was properly positioned and well supported with the retina. All 61 patients cleared a follow-up window of 1-36 months with no reports of important changes in their visual acuity. Among the patients, 91.8% reported normal intraocular pressure, the retinal reattachment rate reached 100%, and the eyeball atrophy control rate reached 100%. There was no report of rupture of the FCVB, allergies to silicone, intraocular infection, intraocular hemorrhage, silicone oil emulsification, or sympathetic ophthalmia. CONCLUSIONS: Foldable capsular vitreous bodies (FCVBs) designed to mimic natural vitreous bodies are suitable as long-term ocular implants that can provide sustained support for the retina without the need for any special postoperative postures. Their barrier function may effectively prolong the retention time of the tamponade and prevent various complications caused by direct contact of the eye tissues with the tamponade.

Tuning the morphology of Cu-MOFs nanostructures for sensitive ascorbic acid sensing.

Metal organic frameworks are an attractive platform to develop fascinating electrocatalysts for the oxidation of ascorbic acid (AA), and their different morphologies have been hinted in literature to impact their sensing performance. In this work, by varying the reaction medium of metal source and organic ligand, copper 2-hydroxybenzene-1,4-dicarboxylate (CuBDC-OH) nanosheets (NSs), nanorods (NRs) and bulk were generated. Thereinto, CuBDC-OH-NSs displayed the highest sensitivity of 151.99µA mM-1cm-2in the linear range of 12-1074µM, which is 1.5 times greater than that of CuBDC-OH NRs and 3.5 times greater than that of CuBDC-OH bulk. The electrochemical analyzes manifested that the superiority of nanosheets originated from higher oxidative current, larger electrochemical active surface area and lower charge transfer resistance, which enabling the efficient electro-oxidation of AA. Additionally, satisfactory selectivity, stability and reproducibility were obtained.

Monorail External Fixation in Humeral Lengthening and Deformity Correction with Glenohumeral Reduction: A Case Report.

CASE: External fixation has an established use in humeral lengthening and deformity correction. An 11-year-old boy presented with left-sided humeral shortening, proximal varus deformity, and glenohumeral dislocation. Anatomical and functional corrections were achieved through innovative monorail external fixation, including bifocal osteotomies of the surgical neck and diaphysis proximal to the deltoid insertion. CONCLUSION: Monorail external fixation with bifocal osteotomies of the surgical neck and diaphysis proximal to the deltoid insertion may improve the anatomical and functional outcomes in humeral shortening and proximal varus deformity with glenohumeral dislocation.

Hierarchical copper-based metal-organic frameworks nanosheet assemblies for electrochemical ascorbic acid sensing.

Noninvasive human health monitoring requires the development of efficient electrochemical sensors for the quantitative analysis of infinitesimal biomolecules. In this work, we reported a novel hierarchical nanosheet assemblies (HSA) of copper-based metal-organic frameworks (MOFs) as an electrochemical sensor for ascorbic acid (AA) detection. Copper 1,4-benzenedicarboxylate (CuBDC) HSA was constructed by three steps of in situ growth on stone paper, including hydrolysis, anion exchange, and heteroepitaxy growth. The monodispersed two-dimensional MOFs nanosheet units were aligned in an orderly manner and arranged into three-dimensional hierarchical assemblies. The CuBDC HSA-based AA sensor displayed a high sensitivity of 396.8 µA mM-1 cm-2 and a low detection limit of 0.1 µM. Excellent selectivity, stability and reproducibility were also obtained. Benefiting from the advantages of ultrathin nanosheets and nature-inspired hierarchy, this unique architecture facilitated reactant dispersion and maximized the accessible active sites and charge-transport capability and thus had superior catalytic ability for the electro-oxidation of ascorbic acid compared to bulk MOFs. Moreover, the CuBDC HSA sensor performed AA level detection in juice samples with acceptable accuracy and verified the feasibility for sweat AA sensing. This novel MOFs architecture holds great potential as an electrochemical sensor to detect AA for noninvasive human health monitoring in the future.

Triplanar osteotomy combined with proximal tibial transverse transport to accelerate healing of recalcitrant diabetic foot ulcers.

BACKGROUND: Management of recalcitrant diabetic foot ulcers remains challenging. Tibial transverse transport (TTT) is an effective method for enhancing the healing of foot ulcers. This retrospective study reports a novel triplanar osteotomy in the tibia and assesses the clinical outcomes of TTT for diabetic foot ulcers. METHODS: Fifty-nine patients with recalcitrant diabetic foot ulcers were divided into the TTT (32 patients) and control (27 patients) groups. In the TTT group, the patients underwent triplanar osteotomy of the proximal tibia, followed by 2 weeks of medial distraction and 2 weeks of lateral distraction. In the control group, the patients received conventional management, including debridement, revascularization, and reconstruction. Ulcer healing and healing time, amputation, recurrence, and complications were assessed at an 18-month follow-up visit. Computed tomography angiography (CTA) was used to evaluate vessel changes in the lower limbs of patients in the TTT group. RESULTS: The TTT group was superior to the control group in the healing rate (90.6% [29/32] vs. 66.7% [18/27]) and the healing time (4.6 ± 1.7 months vs. 7.4 ± 2.5 months), respectively. The proportions of amputation and recurrence in the TTT group were lower than that in the control group, without statistical difference. After triplanar osteotomy and transverse distraction, CTA demonstrated an increase in small vessels in the wound and ipsilateral limb. All patients achieved satisfactory union of the osteotomized bone fragment after removal of the external fixator. CONCLUSIONS: Triplanar osteotomy combined with proximal tibial transverse distraction accelerates wound healing and limb salvage caused by severe and recalcitrant diabetic foot ulcers. Triplanar osteotomy not only increases the bone contact area, which is beneficial for rapid bone reconstruction, but also preserves the vascularization of the bone fragment and substantially facilitates capillary angiogenesis during distraction. These results suggest that triplanar osteotomy followed by tibial transverse distraction is an effective method for treating diabetic foot ulcers.

Functional Characterization of Two RNA Methyltransferase Genes METTL3 and METTL14 Uncovers the Roles of m6A in Mediating Adaptation of Plutella xylostella to Host Plants.

N6-methyladenosine (m6A) is one of the major epigenetic modifications in eukaryotes. Although increasing functions of m6A have been identified in insects, its role in Plutella xylostella L. for host plant adaptation remains unclear. In the current study, we show that the m6A content of P. xylostella was relatively low in different developmental stages and tissues, with no significant differences. Two RNA methyltransferase genes, PxMETTL3 (methyltransferase-like 3) and PxMETTL14 (methyltransferase-like 14), were identified and characterized. PxMETTL3 could be transcribed into two transcripts, and PxMETTL14 had only one transcript; both of these genes were highly expressed in egg and adult stages and reproductive tissues. The CRISPR/Cas9-mediated knockout of PxMETTL3 (ΔPxMETTL3-2) or PxMETTL14 (ΔPxMETTL14-14) confirmed their function in m6A installation into RNA. Furthermore, upon transfer from an artificial diet to the host plant, the mutant strains were affected in terms of larval and pupal weight or adult emergence rate, while the wildtype (WT) strain did not exhibit any difference. In addition, the fecundity and egg hatching rate of the WT strain decreased significantly, whereas only the ΔPxMETTL14-14 mutant strain displayed significantly decreased fecundity. There seemed to be a tradeoff between the stress adaptation and reproduction in P. xylostella mediated by m6A modification. During host transfer, the expression of PxMETTL14 was consistent with the change in m6A content, which implied that PxMETTL14 could respond to host plant defense effectively, and may regulate m6A content. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the differentially expressed transcripts with changes in m6A levels revealed that the potential functions of m6A-related genes may be involved in steroid biosynthesis for larval performance and metabolic pathways for adult reproduction. Overall, our work reveals an epigenetic regulation mechanism for the rapid adaptation of P. xylostella to variations in the host environment.

Influence on emergency digit replantation and outcome assessment after COVID-19 virus nucleic acid testing normalization.

Objective: The study aims to compare the implementation and prognosis of emergency digit replantation surgery before and after normalized corona virus disease 2019 (COVID-19) nucleic acid testing for patients taking emergency operation and to explore the influence of normalized COVID-19 nucleic acid testing on replantation surgery. Method: Normalized COVID-19 nucleic acid testing for patients taking emergency operation has been carried out since 1 August 2021 at our hospital, which means each patient who needs emergency surgical treatment has to obtain either positive or negative results of COVID-19 nucleic acid before entering the operating room. This research reviewed and compared the prognosis of the injured extremity that had emergency severed digit replantation between June and September 2021, at the Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, and analyzed the impact of normalized COVID-19 nucleic acid testing on the outcome of the replanted fingers of different severity using disability of arm-shoulder-hand (DASH) and hand injury severity scoring (HISS) scoring systems. Results: A total of 54 cases with 74 severed replanted phalanges were included replanted by the research group between 1 August and 30 September 2021, without any COVID-19 suspected/confirmed case detected. Compared with previous period (1 June to 31 July, 2021), although the interval between emergency visits and emergency replantation did increase significantly after normalized COVID-19 nucleic acid testing [(3.83 ± 0.94) to (1.77 ± 0.67) h, P < 0.05], we observed no significant difference in the improvement rate of the DASH scoring of the disabled upper extremity 3-month postoperatively (P = 0.538) nor in the complication rate (P = 0.344). Moreover, there was no significant difference in the improvement rate of the DASH scoring of the disabled upper extremity 3-month postoperatively in patients with different traumatic severities before and after normalized COVID-19 nucleic acid testing (moderate P = 0.269, severe P = 0.055, major P = 0.149). Conclusion: Despite the preoperative delay, the policy of COVID-19 nucleic acid testing normalization does not have explicit influence on the short-term outcomes of emergency digit replantation surgery. With this evidence, microsurgeons could pay attention to the patients' anxiety and spend more effort in comforting them during the prolonged preoperative wait. These insights may have implications for other emergency department resource management whenever a social crisis occurs.

Upregulation of E-cadherin in bronchoalveolar lavage fluid-derived exosomes in patients with lung cancer.

BACKGROUND: Lung cancer features extremely high rates of morbidity and mortality. Bronchoalveolar lavage fluid (BALF), obtained by bronchoscopy and bronchoalveolar perfusion, can provide information on the cellular components of the lung microenvironment to assist with diagnosis and treatment of lung cancer. METHODS: BALF was performed using a flexible bronchofiberscope. Exosomes were collected by ultracentrifugation. ELISA detected the amount of E-cadherin. Transmission electron microscopic, ELISA and WB were conducted to identify the existence of the exosomes. Transwell and Wound healing assays were used to detect the ability of migration and invasion. RESULTS: We identified the existence of exosomes in BALF. Furthermore, we observed larger amounts of E-cadherin in the BALF obtained from patients with lung cancer than in the control obtained from the healthy side of pneumonia. Exosomes from lung cancer groups promoted the migration and invasion of A549 cancer cells. CONCLUSION: The exosomes from lung cancer BALF promoted the migration and invasion of A549 cancer cells by carrying E-cadherin. E-cadherin on the surface of exosomes may act through a VE-cadherin dependent mechanism and induce lung cancer metastasis.

Inhibition of Obtusifolin on retinal pigment epithelial cell growth under hypoxia.

AIM: To explore the effect of Obtusifolin on retinal pigment epithelial cell growth under hypoxia. METHODS: In vitro chemical hypoxia model of ARPE-19 cells was established using cobalt chloride (CoCl2). Cell viability was tested by cell counting kit-8 (CCK-8) assay. Western blot and real-time quantitative polymerase chain reaction were applied to detect proteins and mRNAs respectively. Flow cytometry was used to examine the cell cycle. Secretion of vascular endothelial growth factor (VEGF) was tested by using enzyme linked immunosorbent assay (ELISA). RESULTS: Under the chemical hypoxia model established by CoCl2, hypoxia inducible factor-1α (HIF-1α) mRNA and protein levels was up-regulated. Cell viability was increased and the proportion of S phase was higher. Obtusifolin could reduce cell viability under hypoxic conditions and arrest cells in G1 phase. Obtusifolin reduced the expression of Cyclin D1 and proliferating cell nuclear antigen (PCNA) in the hypoxic environment and increased the expression of p53 and p21. The levels of VEGF, VEGFR2 and eNOS proteins and mRNA were significantly increased under hypoxia while Obtusifolin inhibited the increasing. CONCLUSION: Obtusifolin can inhibit cell growth under hypoxic conditions and down-regulate HIF-1/VEGF/eNOS secretions in ARPE-19 cells.

New Development in Traditional Chinese Medicine: Symbolism-Digit Therapy as a Special Naturopathic Treatment.

Traditional Chinese medicine (TCM) grew out of traditional Chinese culture. For example, the eight-diagram symbol is composed of the Yang and the Yin. According to oriental philosophy, everything in the universe has a two-sided property, namely an image (or symbol) and a number (or digit). This paper introduces the new concept and historical background of symbolism-digit therapy (SDT), which is a natural therapy for the treatment of various kinds of diseases. SDT is of TCM heritage, which can be traced back to the ancient publications such as Yi Jing, and this heritage has been incorporated into modern development and practices. The successful treatments using SDT presented in this paper use formulas and/or prescriptions in accordance with TCM. All materials presented in this paper come from first-hand clinical observations, which are supported by TCM theories. Effects of SDT treatments are straightforward and worthy of broader and deeper investigation. SDT and other relevant therapies motivate the further exploration of the essence of TCM to improve the understanding of TCM principles.

Intestine-derived Clostridium leptum induces murine tolerogenic dendritic cells and regulatory T cells in vitro.

Patients with autoimmune and allergic diseases frequently present with reduced numbers and functionally impaired regulatory T cells (Tregs) and/or tolerogenic dendritic cells (tDCs). tDC-mediated regulation of Treg proliferation (numbers) and activation is crucial to establishing and maintaining an appropriate level of immune tolerance. Colonic colonization of Clostridium spp. is associated with accumulation of Tregs, which inhibits development of inflammatory lesions. To investigate whether infection with the Clostridium leptum sp. can specifically induce Tregs and/or tDCs bone marrow-derived dendritic cells were cultured in the presence or absence of C. leptum then co-cultured with CD4(+)CD25(-) T cells or not. Changes in tDC numbers, Treg numbers, percentages of T cell subsets, and expression of cytokines related to Tregs (IL-10 and transforming growth factor-beta (TGF-ß1)), DCs (IL-12p40 and IL-6) and effector T cells (IFN-γ, IL-4, IL-5, IL-13, and IL-17A) were measured. In the co-culture system, C. leptum-stimulated tDCs were able to increase the percentage and total number of Tregs attenuate activation of T helper cells (Th1, Th2, and Th17), and decrease the amount of secreted IL-4, IL-5, IL-13, IFN-γ and IL-17A. Thus, C. leptum exposure can induce the tDC-mediated stimulation of Tregs while disrupting the immune inflammatory response mediated by Th1, Th2 and Th17 cells.

[Prevalence and causes of blindness and moderate and severe visual impairment among adults aged 50 years or above in Longyao County of Hebei Province:the China Nine-Province survey].

OBJECTIVE: To investigate the prevalence and causes of blindness and moderate and severe visual impairment among adults aged ≥ 50 years in Longyao County, Hebei Province, China. METHODS: It was a population-based cross-section study.Geographically defined cluster sampling was used in randomly selecting 5527 individuals aged ≥ 50 years in Longyao County. The survey was preceded by a pilot study where operational methods were refined and quality assurance evaluation was carried out. All participants were enumerated through village registers followed door-to-door visits.Eligible individuals were invited to receive visual acuity measurement and eye examination. Prevalence of blindness and moderate and severe visual impairment was calculated according to different age, gender or education. And the reasons of blindness were analyzed.Statistical analyses were performed using Stata/SE Statistical Software, release 9.0. Chi-square test was used to investigate the association of age, gender and education with presenting and best corrected visual acuity. RESULTS: Five thousands five hundreds and twenty-seven individuals were enumerated and 5051 persons were examined, the response rate was 91.39%. Based on the criteria of World Health Organization visual impairment classification in 1973, the prevalence of blindness and moderate and severe visual impairment defined as best corrected visual acuity was 1.05% (53/5051) and 3.46% (175/5051) respectively. The prevalence of blindness and moderate and severe visual impairment defined as presenting visual acuity was 1.48% (75/5051) and 7.94% (401/5051) respectively. The prevalence of blindness and moderate and severe visual impairment was higher in aged (trend χ(2) = 897.27, P = 0.000) , female (χ(2) = 30.32, P = 0.000), illiterate (trend χ(2) = 83.20, P = 0.000) persons. Cataract was still the first leading cause of blindness. Un-corrected refractive error also was the main cause of visual impairment. CONCLUSIONS: The prevalence of blindness and moderate and severe visual impairment is relatively lower among China Nine Province Survey. The first leading cause of blindness and visual impairment is still cataract.

Silencing IL-23 expression by a small hairpin RNA protects against asthma in mice.

To determine the impact of IL-23 knockdown by RNA interference on the development and severity of ovalbumin (OVA)-induced asthmatic inflammation, and the potential mechanisms in mice, the IL-23-specific RNAi-expressing pSRZsi-IL-23p19 plasmid was constructed and inhaled into OVA-sensitized mice before each challenge, as compared with that of control mice treated with alum or budesonide. Inhalation of the pSRZsi-IL-23p19, significantly reduced the levels of OVA-challenge induced IL-23 in the lung tissues by nearly 75%, determined by RT-PCR. In addition, knockdown of IL-23 expression dramatically reduced the numbers of eosinophils and neutrophils in BALF and mitigated inflammation in the lungs of asthmatic mice. Furthermore, knockdown of IL-23 expression significantly decreased the levels of serum IgE, IL-23, IL-17, and IL-4, but not IFNgamma, and its anti-inflammatory effects were similar to or better than that of treatment with budesonide in asthmatic mice. Our data support the notion that IL-23 and associated Th17 responses contribute to the pathogenic process of bronchial asthma. Knockdown of IL-23 by RNAi effectively inhibits asthmatic inflammation, which is associated with mitigating the production of IL-17 and IL-4 in asthmatic mice.

Molecular dynamics simulations reveal the disparity in specific recognition of GCC-box by AtERFs transcription factors super family in Arabidopsis.

Arabidopsis ethylene responsive element binding factors (AtERFs) form a transcription factor super family. While the functions of most AtERFs are unknown, a number of AtERFs appear to be involved in regulation of stress-related genes through their DNA binding domains (DBD), namely ERF domains, which recognize a consensus motif GCC-box at the regulatory region. In this study, molecular dynamics simulations were performed on the four ERF domain-GCC-box complexes, AtERF1, AtERF4, AtEBP and CFBF1, to determine disparity in specific binding to the GCC-box by the AtERFs. Our results suggested that three amino acid residues Arg29, Glu39 and Arg41, played a vital role in direct readout of DNA. The position of the consensus sequence GCCGCC has an intrinsic disparity on binding with ERF domains. The third C, fourth G and the last C in the GCC motif was compulsory for recognition by ERF domains. Our results provide structural evidence for a sequence-dependent recognition mechanism for AtERFs.