Chromatographic Purification and Polishing of AAV Particles.

The production of Adeno-associated virus (AAV) vectors in the lab setting has typically involved expression in adherent cells followed by purification through ultracentrifugation in density gradients. This production method is, however, not easily scalable, presents high levels of cellular impurities that co-purify with the virus, and results in a mixture of empty and full capsids. Here we describe a detailed AAV production protocol that overcomes these limitations through AAV expression in suspension cells followed by AAV affinity purification and AAV polishing to separate empty and full capsids, resulting in high yields of ultra-pure AAV that is highly enriched in full capsids.

In situ study on articular cartilage degeneration in simulated microgravity by HOF-ATR-FTIR spectroscopy.

Fourier transform infrared spectroscopy (FTIRS) can provide rich information on the composition and content of samples, enabling the detection of subtle changes in tissue composition and structure. This study represents the first application of FTIRS to investigate cartilage under microgravity. Simulated microgravity cartilage model was firstly established by tail-suspension (TS) for 7, 14 and 21 days, which would be compared to control samples. A self-developed hollow optical fiber attenuated total reflection (HOF-ATR) probe coupled with a FTIR spectrometer was used for the spectral acquisition of cartilage samples in situ, and one-way analysis of variance (ANOVA) was employed to analyze the changes in the contents of cartilage matrix at different stages. The results indicate that cartilage degenerates in microgravity, the collagen content gradually decreases with the TS time, and the structure of collagen fibers changes. The trends of proteoglycan content and collagen integrity show an initial decrease followed by an increase, ultimately significantly decreasing. The findings provide the basis for the cartilage degeneration in microgravity with TS time, which must be of real significance for space science and health detection.

Parotid Gland Suspension Combined with Rhytidectomy to Recontour the Lower Face and Neck Enlargement Caused by Benign Parotid Hypertrophy.

BACKGROUND: Lower face and neck aging and facial shape changes are usually caused by soft tissue descent. During lower face and neck enlargement, parotid hypertrophy is an important influence factor of morphologic changes in the periauricular regions. Most plastic surgery techniques greatly improve the shape of the lower face and neck, but ignore the manipulation of the parotid gland. We aimed to resolve the lower face and neck enlargement caused by parotid hypertrophy. Thus, we performed parotid gland suspension combined with rhytidectomy to improve the lower face and neck contour. METHODS: This retrospective study recruited 22 patients who underwent parotid gland suspension combined with rhytidectomy from 2012 to 2020. The evaluation of patient outcomes was performed by selecting items from the FACE-Q scale. Surgical procedures involved parotid gland exposure and parotid fascia suspension. RESULTS: Patient-perceived age appraisal indicated a younger appearance with follow-ups for approximately 3-10 year after the surgery (mean ± standard deviation - 5.53 ± 3.67). Patients had a high satisfaction level for facial appearance (67.15 ± 16.84), lower face (74.69 ± 21.22), and contour of the parotid gland areas of the neck (65.76 ± 23.62). The lower face and neck contours were narrowed and tightened. The parotid gland area showed a remarkable improvement after surgery. CONCLUSIONS: Parotid gland suspension combined with rhytidectomy obtained an outstanding improvement. This method can achieve a better lower face and neck contour, especially in patients with benign parotid hypertrophy or lower face and neck enlargement in periauricular regions. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Engineered Shape-Morphing Transitions in Hydrogels Through Suspension Bath Printing of Temperature-Responsive Granular Hydrogel Inks.

4D printing of hydrogels is an emerging technology used to fabricate shape-morphing soft materials that are responsive to external stimuli for use in soft robotics and biomedical applications. Soft materials are technically challenging to process with current 4D printing methods, which limits the design and actuation potential of printed structures. Here, a simple multi-material 4D printing technique is developed that combines dynamic temperature-responsive granular hydrogel inks based on hyaluronic acid, whose actuation is modulated via poly(N-isopropylacrylamide) crosslinker design, with granular suspension bath printing that provides structural support during and after the printing process. Granular hydrogels are easily extruded upon jamming due to their shear-thinning properties and their porous structure enables rapid actuation kinetics (i.e., seconds). Granular suspension baths support responsive ink deposition into complex patterns due to shear-yielding to fabricate multi-material objects that can be post-crosslinked to obtain anisotropic shape transformations. Dynamic actuation is explored by varying printing patterns and bath shapes, achieving complex shape transformations such as 'S'-shaped and hemisphere structures. Furthermore, stepwise actuation is programmed into multi-material structures by using microgels with varied transition temperatures. Overall, this approach offers a simple method to fabricate programmable soft actuators with rapid kinetics and precise control over shape morphing.

Enhanced production of recombinant coxsackievirus A16 using a serum-free HEK293A suspension culture system for bivalent enterovirus vaccine development.

Coxsackievirus A16 (CVA16) is one of the primary pathogens that causes hand, foot, and mouth disease (HFMD) in young children. In previous studies, CVA16 vaccine development has encountered several challenges, such as inefficient replication of the CVA16 virus in present culture systems, the induction of only mild neutralizing antibody titers, and neutralizing antibodies induced by certain vaccine candidates that are unable to protect against CVA16 viral challenge. In this study, we constructed a DNA-launched CVA16 infectious clone (CVA16ic) based on the genomic sequence of the CVA16 N5079 strain to minimize interference from viral quasispecies. The biochemical properties of this CVA16ic strain were similar to those of its parental strain. Serum-free HEK293A suspension cells, which produced higher virus titers than Vero cells, were demonstrated to improve CVA16 production yields. In addition, our study showed that inactivated EV-A71 antigens could enhance the immunogenicity of inactivated CVA16 mature/full particles (F-particles), suggesting that a bivalent CVA16 and EV-A71 vaccine may be an effective strategy for CVA16 vaccine development. These findings are expected to provide novel strategies and accelerate the development of bivalent HFMD vaccines.

Alcohol use disorder and fitness to drive: Discrepancies between health professionals' evaluations and objective measures of alcohol use and cognitive functioning.

Aims: In this study, we investigated if health professionals' evaluations of driving ability corresponded with measures of severity of alcohol use and measures of cognitive functions necessary for safely driving a car. Methods: A total of 90 participants from a multicentre study were included. Participants were categorised into three groups: (1) the group judged fit to drive (FIT); (2) the group judged not fit to drive (UNFIT); and (3) the group who had lost their driver's licence due to legal sanctions (LEGAL). The participants' AUDIT scores, earlier treatment episodes and results from neuropsychological tests of reaction time, attention and visuospatial ability were included in the analyses. Results: We found a significant difference in the severity of alcohol use disorder (AUD) and visuospatial abilities between the FIT and UNFIT groups. Half of the UNFIT group had at least mild visuospatial difficulties, compared to only a quarter in the FIT group. There were no group differences in reaction time or attentional measures. The LEGAL group had more severe AUD than the other groups. Conclusion: The FIT group did not perform differently from the UNFIT group on attention and reaction time measures. The UNFIT group had more visuospatial impairments, but even half of this group had normal scores. It is uncertain whether the differences between the two groups are of practical significance. The quality of health professionals' evaluations may be questioned, and the results highlight the need for more reliable and valid criteria for doing fitness to drive evaluations.

A comparative characterization analysis of electrode materials for optimal deposition on copper by advancing electrical discharge coating.

This research explores the efficiency of Electrical Discharge Coating (EDC) as an advanced surface modification technique using powder suspension, conventional electrodes, and 3D printed electrodes (3DPE), focusing on Ti6Al4V alloys. Conventional Ti electrodes resulted in 100% titanium content with a thickness of 110 µm, while the powder suspension coating was inadequate. The use of 3D printed electrodes showed promising results, with uniform deposition of 61.20 µm thickness and enhanced surface properties under 10 A current. Further analysis of 3D printed Ti6Al4V EDC revealed a higher Ti percentage of 44.20%, TiC formation of 84.17%, and enhanced microhardness with lower surface roughness. Thorough optimization of 3D printed electrode EDC has been undertaken. The study suggests the potential replacement of traditional electrodes such as powder metallurgy with 3D printed counterparts, advancing surface modification techniques and opening new avenues for materials engineering and manufacturing processes.

Development and integration of a continuous horizontal belt filter into drug production procedure.

In the pharmaceutical industry, filtration is traditionally carried out in batch mode. However, with the spread of continuous technologies, there is an increasing demand for robust continuous filtration strategies suitable for processing suspensions produced in continuous crystallizers. Accordingly, this study aimed to investigate a lab-scale horizontal conveyor belt filtration approach for pharmaceutical separation purposes for the first time. The newly developed continuous horizontal belt filter (CHBF) was tested under different systems (microcrystalline cellulose (MCC)/water, lactose/ethanol and acetylsalicylic acid (ASA)/water) and diverse conditions. Filtration was robust using a well-defined unimodal particle size distribution MCC in water system, where the residual moisture content varied within narrow limits of 45-52% independently from the process conditions. Besides, the residual moisture content highly depended on the applied solvent and particle size. It could be reduced to below 2% by processing the suspensions of either a volatile solvent (lactose in ethanol) or an aqueous slurry of a large particle size ASA. Finally, the CHBF was connected to a mixed suspension mixed product removal (MSMPR) or a plug flow crystallizer (PFC). The residual moisture content of the CHBF-filtered ASA product and operation characteristics (onset of steady-state) were evaluated in both continuous crystallizer-filter systems. The MSMPR-CHBF system operated with a longer startup period. The size of the in situ-produced crystals was of a similar order magnitude in both systems, resulting in a similar residual moisture content (around 20%). Overall, the tested continuous filter was robust, did not modify the crystal morphology in the examined experimental range, and could be effectively integrated with continuous crystallizers.

Salvage Reconstruction of Composite Defects of the Anterior Mandible, Floor of Mouth, and Lip.

Anterior mandible defects result in loss of support for the tongue, floor of the mouth and lower lip, resulting in impairment of airway, feeding, and speech. We treated four patients with these "Andy Gump" deformities. Reconstruction was performed with two free flaps: a fibula osteocutaneous flap for the anterior mandible and floor of the mouth, and a soft tissue free flap for the lip, chin, and anterior neck. The lower lip was suspended cranially with fascia or tendon grafts ± mini-temporalis turndown flaps. All flaps survived completely. All patients were tube feed-dependent before surgery; they all resumed an oral diet. All tracheostomies were decannulated. Lip competence was restored as evidenced by cessation of drooling. Speech improved from unintelligible to intelligible with frequent repetitions. Objective assessment was performed with the functional intraoral Glasgow scale; the mean FIGS score improved from 3.25 (range 3-4) to 11 (range 9-13). We conclude that composite anterior mandible and tongue defects have large tissue requirements that require multiple free flaps. Reconstruction leads to significant improvement in function.

Mapping Microplastic Movement: A Phase Diagram to Predict Nonbuoyant Microplastic Modes of Transport at the Particle Scale.

Microplastics pose numerous threats to aquatic environments, yet understanding their transport mechanisms remains limited. Drawing from natural sediment research provides valuable insights to address this knowledge gap. One key dimensionless number used to describe sediment transport is the transport stage, referring to the ratio between the flow shear velocity and the particle settling velocity. However, variations in physical properties, such as shape and density, raise concerns about the applicability of existing sediment transport theories to microplastics. To address this challenge, we employed a physical modeling approach, examining 24 different nonbuoyant microplastic particles in a turbulent open channel flow. Utilizing 3D particle tracking, a total of 720 trajectories were recorded and analyzed. Microplastic particles exhibited transport modes akin to natural sediments, including rolling/sliding, saltation, and suspension. The transport stage strongly correlated with these modes, as well as with the mean forward velocity and mean position in the water column. Notably, particle shape emerged as a critical factor influencing transport dynamics. Due to their lower settling velocity, fibers tended to stay closer to the water surface with lower forward velocities compared to spheres. Based on the laboratory results, a new phase diagram for microplastics is introduced analogous to an existing diagram for sediments.

Impact of seasonal changes and environmental conditions on suspended and inhalable microplastics in urban air.

Microplastics (MPs) are ubiquitous environmental pollutants extensively detected in atmospheric environments. Airborne MPs have raised concerns due to their transport and potential health risks of inhalation exposure. However, the factors influencing airborne MPs, particularly their concentrations and shapes suspended in urban air, remain unclear. We investigated MPs in total suspension particles with one-year measurements in Taipei City and identified their features using Nile Red staining combined with fluorescence microscopy and micro-Fourier transform infrared (µFTIR) spectroscopy. This study quantified the mean number concentration of total MPs as approximately 6.0 #/m³. We observed that MP abundance varied seasonally, with higher levels in the warm season than in the cold. A similar trend was noted for polymer types. Fragment-like MPs were the predominant shape, mainly found in polystyrene (PS), polyethylene (PE), and polypropylene (PP), while fibrous MPs, detected mostly as polyethylene terephthalate (PET) and polyamide (PA), were primarily observed at sizes greater than 300 µm. Both fiber and fragment-like MPs were positively associated with particle mass concentration, temperature, ultraviolet (UV) index, and wind speed, but negatively correlated with relative humidity and rainfall. Fibrous MPs were more affected by environmental factors than fragment-like MPs. Meteorological changes significantly influenced suspended MPs more than human activity within the city.

Endophyte-based fungal elicitors for enhanced production of valepotriates and sesquiterpenoids in leaf cell suspension cultures of Valeriana jatamansi Jones.

AIMS: The immense therapeutic value of Valeriana jatamansi is attributed to the presence of bioactive secondary metabolites (valepotriates and sesquiterpenoids). Its over-exploitation in wild habitats resulted in extensive depletion, necessitating alternative approaches to produce its therapeutic metabolites. This study sought to assess the ability of endophytes of V. jatamansi to boost the biosynthesis of secondary metabolites in the leaf-cell suspension (LCS) culture of V. jatamansi. METHODS AND RESULTS: A total of 11 fungal endophytes were isolated from the rhizomes of V. jatamansi. Isolated endophytes were found to belong to phylum Ascomycota, Basidiomycota, and Mucoromycota. Supplementation of extracts of endophyte Phaeosphaeriaceae sp. VRzFB, Mucor griseocyanus VRzFD, Penicillium raistrickii VRzFK, and Penicillium sajarovii VRzFL in the LCS culture of V. jatamansi increased the fresh cell biomass by 19.6%-39.1% and dry cell biomass by 23.4%-37.8%. Most of the endophytes' extract could increase the content of valepotriates (26.5%-76.5% valtrate and 40.5%-77.9% acevaltrate) and sesquiterpenoids (19.9%-61.1% hydroxyl valerenic acid) in LCS culture. However, only two endophytes, Irpex lacteus VRzFI and Fusarium oxysporum VRzFF, could increase the sesquiterpenoids acetoxy valerenic acid (36.9%-55.3%). In contrast, some endophytes' extracts caused negative or no significant effect on the cell biomass and targeted metabolites. Increased secondary metabolites were corroborated with increased expression of iridoid biosynthesis genes in LCS culture. Production of H2O2 and lipid peroxidation was also varied with different endophytes indicating the modulation of cellular oxidative stress due to endophyte elicitors. CONCLUSIONS: The results suggest the distinct effect of different fungal endophytes-extract on LCS culture, and endophytes can serve as biotic elicitors for increasing the secondary metabolite production in plant in vitro systems.

Generational Diet-Induced Obesity Remodels the Omental Adipose Proteome in Female Mice.

Obesity, a complex condition that involves genetic, environmental, and behavioral factors, is a non-infectious pandemic that affects over 650 million adults worldwide with a rapidly growing prevalence. A major contributor is the consumption of high-fat diets, an increasingly common feature of modern diets. Maternal obesity results in an increased risk of offspring developing obesity and related health problems; however, the impact of maternal diet on the adipose tissue composition of offspring has not been evaluated. Here, we designed a generational diet-induced obesity study in female C57BL/6 mice that included maternal cohorts and their female offspring fed either a control diet (10% fat) or a high-fat diet (45% fat) and examined the visceral adipose proteome. Solubilizing proteins from adipose tissue is challenging due to the need for high concentrations of detergents; however, the use of a detergent-compatible sample preparation strategy based on suspension trapping (S-Trap) enabled label-free quantitative bottom-up analysis of the adipose proteome. We identified differentially expressed proteins related to lipid metabolism, inflammatory disease, immune response, and cancer, providing valuable molecular-level insight into how maternal obesity impacts the health of offspring. Data are available via ProteomeXchange with the identifier PXD042092.

Development and evaluation of amine-functionalized ß-cyclodextrin grafted starch as a natural flocculant for turbidity removal in water treatment.

Recently, biopolymers have been used as coagulants/flocculants due to their biodegradability, low cost, and renewability. In this study, an environmentally friendly amine-functionalized starch-based flocculant was successfully prepared. Initially, ß-cyclodextrin was grafted onto the starch backbone to increase the number of hydroxyl groups, and this composite was named CD-starch. Subsequently, in order to introduce cationic properties and enhance effective flocculation, CD-starch was modified using amine functional groups. The surface functional groups were engineered by introducing different amine to CD-starch ratios (0.5:1, 1:1, 2:1 w/w), named A-CD-starch 0.5, 1 and 2, respectively. Following the characterization of the synthesized substrate, its performance in the flocculation process of a kaolin suspension was investigated. The effects of different parameters, including pH, flocculant dosage, and initial turbidity on wastewater turbidity removal, was investigated. The results showed that a higher ratio of amine to CD-starch leads to a better amination reaction due to the greater availability of nitrogen for alkylation. Jar experiments showed that for initial turbidities of 50, 150 and 300 NTU, the appropriate doses of flocculant were 0.070, 0.085 and 0.130 mg/mL, respectively. For these initial turbidities, the maximum turbidity removal was achieved 80.1 %, 92 %, and 97.8 %, respectively. This work provides an innovative natural flocculant based on starch which can effectively treat turbid wastewaters.

Management of the Nasal Valve in Facial Paralysis.

Facial paralysis significantly impacts the form and function of patients. Assessment of the face in zones is important to ensure no functional area of the face is neglected. Nasal valve compromise in patients with facial paralysis, for example, is often overlooked yet should be addressed to correct nasal obstruction. In flaccid facial paralysis, inferomedial displacement of the alar base and lateral nasal sidewall insufficiency contribute to nasal valve compromise. For surgical candidates, static suspension of the nasal valve in a superolateral vector is an ideal technique to address the etiology of nasal obstruction in patients with facial paralysis.

Edible structuring agent shaped via interfacial precipitation on solid template: Crosslinked starch colloidosome.

Water-permeable hollow starch particles alter the rheological behavior of their granular suspensions. However, their thin shells can rupture limiting applications. In this study, we used amaranth starch as building blocks (1 µm) to craft a crosslinked superstructure. Pickering emulsions were used as the templates where starch coated the droplets. Emulsions were heated at 75 °C to induce interpenetration of the polymers followed by precipitation in ethanol to trigger colloidal fusion. Particles were then crosslinked by sodium tri-metaphosphate; hollow particles formed after the interior template was removed by hexane. When canola oil was used, the particles ruptured at pH 11.5 due to the repulsion between the strands. In contrast, palm oil, emulsified at 50 °C, formed a rigid core after cooling, locked the starch at the surface and retained the structure. The crosslinked colloidosomes were larger (89 µm) and exhibited higher viscosity, and stronger stability. Larger particles (>100 µm) were produced using higher templating volume. Gentle centrifugation to harvest the particles kept the shells intact. The hollow structure exhibited jamming transition above 10 w/w%, which could serve as a super-thickener. This work demonstrates that microarchitecture plays a critical role in shaping material functionality.

Numerical study on heat transfer and pressure performance of different suspension nozzles.

The drying process of the lithium battery pole pieces makes extensive use of the suspension nozzle. It is of great significance to study the heat transfer and pressure steady-state characteristics of the suspension nozzle and to select the appropriate nozzle structure for the production of pole pieces. Based on the SST k - ω turbulence model, this article numerically simulates the impact jet process of suspension nozzles with slits, injection holes, and effusion holes. There is a qualitative and quantitative analysis of the distribution of their velocity field, temperature field, local Nusselt number, average Nusselt number, local pressure coefficient, and average pressure coefficient, and the comprehensive performance index of the nozzle is proposed. The results show that when the weight factor of heat transfer performance α is less than 21.61% and the weight factor of pressure performance ß is more than 78.39%, the comprehensive performance of the traditional suspension nozzle with double slits is the best. As the α is increasing, the ß is decreasing. The comprehensive performance of the suspension nozzle with effusion holes is the best. The turbulent intermittence, interaction between neighbouring jets, and edge effects affect the heat transfer and pressure uniformity of the suspension nozzle.

Effectiveness of Amikacin liposome inhalation suspension for refractory Mycobacterium avium complex pulmonary disease at 6 months post initiation.

BACKGROUND: Amikacin  liposome inhalation suspension (ALIS) improved sputum culture conversion rate at 6 months for patients with refractory Mycobacterium avium complex pulmonary disease (MAC-PD) in an international phase 3 trial. Patient characteristics and chest high-resolution CT (HRCT) findings associated with ALIS effectiveness are poorly documented. OBJECTIVE: We aimed to clarify ALIS effectiveness for refractory MAC-PD at 6 months, elucidating associated patient characteristics and chest CT findings. METHODS: We reviewed medical records of 12 patients with refractory MAC-PD for whom ALIS treatment was initiated at Toho University Omori Medical Center from November 2021 through September 2022. All patients demonstrated treatment persistence for at least 3 months. They were divided into culture conversion and non-conversion groups using sputum culture conversion status after 6-month ALIS treatment initiation. Clinical and radiological characteristics were compared. RESULTS: Seven of the 12 patients (58.3%) achieved sputum culture conversion within 6 months. The culture conversion group had shorter pre-ALIS initiation treatment duration [21 months (16-25) vs. 62 months (32-69); p = 0.045]; lower cavitary lesion incidence on HRCT (28.6% vs. 100%; p = 0.028); and fewer clarithromycin (CLA)-resistant strains [0/7 (0%) vs. 3/5 (60%); p = 0.045]. Chest HRCT findings improved in 4 of 7 (57.1%) and 1 of 5 (20%) patients in the culture conversion and non-conversion groups, respectively. CONCLUSION: ALIS facilitated sputum culture conversion within 6 months in 58.3% of patients with refractory MAC-PD. Sputum culture conversion was significantly more frequent for CLA-susceptible strains and patients with fewer cavitary lesions. Improved CT findings after ALIS did not always correspond to sputum culture conversion.

Fed-batch strategies for intensified rVSV vector production in high cell density cultures of suspension HEK293 cells.

Vesicular stomatitis virus (VSV) has been increasingly demonstrated as a promising viral vector platform. As the interest over this modality for vaccine and gene therapy applications increases, the need for intensified processes to produce these vectors emerge. In this study, we develop fed-batch-based operations to intensify the production of a recombinant VSV-based vaccine candidate (rVSV-SARS-CoV-2) in suspension cultures of HEK293 cells. A feeding strategy, in which a commercial concentrated medium was added to cultures based on cell growth through a fixed cell specific feeding rate (CSFR), was applied for the development of two different processes using Ambr250 modular bioreactors. Cultures operated in hybrid fed-batch/perfusion (FB/P) or fed-batch (FB) were able to sustain infections performed at 8.0 × 106 cells/mL, respectively resulting in 3.9 and 5.0-fold increase in total yield (YT) and 1.7 and 5.6-fold increase in volumetric productivity (VP) when compared with a batch reference. A maximum viral titer of 4.5 × 1010 TCID50/mL was reached, which is comparable or higher than other processes for VSV production in different cell lines. Overall, our study reports efficient fed-batch options to intensify the production of a rVSV-based vaccine candidate in suspension HEK293 cells.

Finite Element Modeling and Calibration of a Three-Span Continuous Suspension Bridge Based on Loop Adjustment and Temperature Correction.

Precise finite element modeling is critically important for the construction and maintenance of long-span suspension bridges. During the process of modeling, shape-finding and model calibration directly impact the accuracy and reliability. Scholars have provided numerous alternative proposals for the shape-finding of main cables in suspension bridges from both theoretical and finite element analysis perspectives. However, it is difficult to apply these solutions to suspension bridges with special components. Seeking a viable solution for such suspension bridges holds practical significance. The Nanjing Qixiashan Yangtze River Bridge is the first three-span suspension bridge in China. To maintain the configuration of the main cable, the suspension bridge is equipped with specialized suspenders near the anchors, referred to as displacement-limiting suspenders. It is the first suspension bridge in China to use displacement-limiting suspenders and their anchorage system. Taking the suspension bridge as a research background, this paper introduces a refined finite element modeling approach considering the effect of geometric nonlinearity. Firstly, based on the loop adjustment and temperature correction, the shape-finding and force assessment of the main cables are carried out. On this basis, a nonlinear finite element model of the bridge was established and calibrated, taking into account factors such as pylon settlement and cable saddle precession. Finally, the static and dynamic characteristics of the suspension bridge were thoroughly investigated. This study aims to provide a reference for the design, construction and operation of the three-span continuous suspension bridge.