Pre-cooling for Reducing Pain from Local Anesthetic Injections for Hemorrhoidectomy: An Open-label, Randomized, Crossover Trial.

Objectives: Excisional hemorrhoidectomy (EH) is sometimes same-day surgery under local anesthesia (LA); however, the LA injection can be painful. We conducted an open-label, crossover, randomised controlled trial to evaluate the efficacy and safety of pre-cooling in reducing pain associated with LA injection. Methods: Patients aged ≥ 20 years undergoing bilateral EH were randomly assigned to 1 of 2 pre-cooling sequences: a cooling-first sequence and a cooling-second sequence. In the first intervention phase, 2 minutes of pre-cooling was applied before LA injection in patients randomized to the cooling-first sequence; patients in the cooling-second sequence were asked to wait for 2 minutes (without pre-cooling) before LA injection. The pre-cooling sequences and the perianal sides targeted for injection were reversed in the second intervention phase. The primary outcome was the visual analogue scale (VAS) rating for pain from LA injection, which was obtained twice for each patient. Adverse events due to pre-cooling (e.g., skin disorders) were documented. Results: Of 114 screened patients, 51 were randomized to the cooling-first (n = 26; analyzed: n = 26) or cooling-second sequence (n = 25; analyzed: n = 25). The 2-minute pre-cooling was completed by 48 patients (94%). VAS scores for LA injection pain decreased significantly with pre-cooling compared to without (difference estimate, -1.71; 95% confidence interval, -2.12 to -1.31; p< 0.001). No adverse events were reported. Conclusions: Two minutes of skin pre-cooling effectively and safely reduces LA injection pain in patients undergoing EH.

Ice recrystallization inhibition activity of pulse protein hydrolysates after immobilized metal affinity separation.

Creating molecules capable of inhibiting ice recrystallization is an active research area aiming to improve the freeze-thaw characteristics of foods and biomedical materials. Peptide mixtures have shown promise in preventing freezing-induced damage, but less is known about the relationship between their amino acid compositions and ice recrystallization inhibition (IRI) activities. In this article, we used Ni2+ immobilized metal affinity chromatography (IMAC) to fractionate pulse protein hydrolysates, created by Alcalase and trypsin, into mixtures lacking and enriched in His, and Cys residues. The aim of this study was to fractionate pulse protein hydrolysates based on their amino acid compositions and evaluate their resulting physicochemical and IRI characteristics. Ni2+ IMAC fractionation induced IRI activity in all of the evaluated soy, chickpea, and pea protein hydrolysates regardless of their amino acid composition. Ni2+ IMAC fractionation produced chemically distinct fractions of peptides, differing by their molecular weights, amino acid composition, and IRI activities. The resulting peptide mixtures' molecular weight, amino acid composition, secondary structure, and sodium ion levels were found to have no correlation with their IRI activities. Thus, we demonstrate for the first time the ability of Ni2+ IMAC fractionation to induce IRI activity in hydrolyzed pulse proteins.

EXPRESS: Feasibility of a Fiber-Dispersive Raman Spectrometer for Biomarker Detection.

Raman spectroscopy is among the top analytical techniques for ultra-low-dense organic matter, crucial to the search for life and analysis of celestial body surfaces in space exploration missions. Achieving the ultimate sensitivity in in situ Raman spectroscopy necessitates a breakthrough in detecting inelastically scattered light. Single-photon detectors (SPD) operating in photon counting mode, which can differentiate between Raman and luminescence responses, are promising candidates for the challenging scientific requirements. Since large SPD arrays are not yet commercially available, a dispersive element can be adapted to a single-pixel detector. By exploiting chromatic dispersion in optical fibers and picosecond-pulsed excitation, we delay the arrivals of different spectral components onto a single-pixel SPD. This method also separates weak Raman signals from stronger luminescence through correlated time-domain measurements. We study the impact of fiber properties and the excitation wavelength of a pulsed laser on the spectral resolution of the fiber-dispersive Raman spectrometer (FDRS). Additionally, we demonstrate the FDRS's potential for studying biomarkers and discuss its feasibility for analyzing inclusions in ice matrices.

Hibernating/Awakening Nanomotors Promote Highly Efficient Cryopreservation by Limiting Ice Crystals.

The disruptions caused by ice crystal formation during the cryopreservation of cells and tissues can cause cell and tissue damage. Thus, preventing such damage during cryopreservation is an important but challenging goal. Here, a hibernating/awakening nanomotor with magnesium/palladium covering one side of a silica platform (Mg@Pd@SiO2) is proposed. This nanomotor is used in the cultivation of live NCM460 cells to demonstrate a new method to actively limit ice crystal formation and enable highly efficient cryopreservation. Cooling Mg@Pd@SiO2 in solution releases Mg2+/H2 and promotes the adsorption of H2 at multiple Pd binding sites on the cell surface to inhibit ice crystal formation and cell/tissue damage; additionally, the Pd adsorbs and stores H2 to form a hibernating nanomotor. During laser-mediated heating, the hibernating nanomotor is activated (awakened) and releases H2, which further suppresses recrystallization and decreases cell/tissue damage. These hibernating/awakening nanomotors have great potential for promoting highly efficient cryopreservation by inhibiting ice crystal formation.

Isolated ipsilateral ptosis associated with ventral midbrain infarction: a case report and literature review.

Documented cases of ipsilateral ptosis caused by midbrain infarction remain rare. Herein, we present a patient with isolated ipsilateral ptosis that was initially considered to be a consequence of myasthenia gravis but was subsequently attributed to ventral midbrain infarction. We also discuss the possible underlying mechanisms; ipsilateral ptosis in our patient was attributed to selective damage of the levator palpebral muscle branch of the oculomotor nerve. The patient was started on aspirin (200 mg once daily) and atorvastatin (40 mg once daily). Improvement in ptosis occurred from day 5 of admission, and the patient was subsequently discharged. Ptosis disappeared 1 month after onset. This report describes an extremely rare case of ventral midbrain infarction presenting with isolated ipsilateral ptosis. Careful examination, including magnetic resonance imaging, is essential in such patients, especially in those with multiple cerebrovascular risk factors.

Study on the key parameters of ice particle air jet ejector structure.

Existing ice particle jet surface treatment technology is prone to ice particle adhesion during application, significantly affecting surface treatment efficiency. Based on the basic structure of the jet pump, the ice particle air jet surface treatment technology is proposed for the instant preparation and utilization of ice particles, solving the problem of ice particle adhesion and clogging. To achieve efficient utilization of ice particles and high-speed jetting, an integrated jet structure for ice particle ejection and acceleration was developed. The influence of the working nozzle position (Ld), expansion ratio (n), and acceleration nozzle diameter ratio (Dn) length-to-diameter ratio (Ln) on the ice particle ejection and acceleration was systematically studied. The structural parameters of the ejector were determined using the impact kinetic energy of ice particles as the comprehensive evaluation index, and the surface treatment test was conducted to verify the results. The study shows that under 2 MPa air pressure, the ejector nozzle parameters of n = 1.5, Dn = 4.0, Ld = 4, and Ln = 0 mm can effectively eject and accelerate the ice particles. The aluminum alloy plate depainting test obtained a larger paint removal radius and resulted in a smoother aluminum alloy plate surface, reducing the surface roughness from 3.194 ± 0.489 µm to 1.156 ± 0.136 µm. The immediate preparation and utilization of ice particles solved the problems of adhesion and storage in the engineering application of ice particle air jet technology, providing a feasible technical method in the field of material surface treatment.

Historical Southern Hemisphere biomass burning variability inferred from ice core carbon monoxide records.

Biomass burning plays an important role in climate-forcing and atmospheric chemistry. The drivers of fire activity over the past two centuries, however, are hotly debated and fueled by poor constraints on the magnitude and trends of preindustrial fire regimes. As a powerful tracer of biomass burning, reconstructions of paleoatmospheric carbon monoxide (CO) can provide valuable information on the evolution of fire activity across the preindustrial to industrial transition. Here too, however, significant disagreements between existing CO records currently allow for opposing fire histories. In this study, we reconstruct a continuous record of Antarctic ice core CO between 1821 and 1995 CE to overlap with direct atmospheric observations. Our record indicates that the Southern Hemisphere CO burden ([CO]) increased by 50% from a preindustrial mixing ratio of ca. 35 ppb to ca. 53 ppb by 1995 CE with more variability than allowed for by state-of-the-art chemistry-climate models, suggesting that historic CO dynamics have been not fully accounted for. Using a 6-troposphere box model, a 40 to 50% decrease in Southern Hemisphere biomass-burning emissions, coincident with unprecedented rates of early 20th century anthropogenic land-use change, is identified as a strong candidate for this mismatch.

Development of a Modular Cryo-Transfer Station for the Side-Entry Transmission Electron Microscope†.

Cryo-transfer stations are essential tools in the field of cryo-electron microscopy, enabling the safe transfer of frozen vitreous samples between different stages of the workflow. However, existing cryo-transfer stations are typically configured for only the two most popular sample holder geometries and are not commercially available for all electron microscopes. Additionally, they are expensive and difficult to customize, which limits their accessibility and adaptability for research laboratories. Here, we present a new modular cryo-transfer station that addresses these limitations. The station is composed entirely of 3D-printed and off the shelf parts, allowing it to be reconfigured to a fit variety of microscopes and experimental protocols. We describe the design and construction of the station and report on the results of testing the cryo-transfer station, including its ability to maintain cryogenic temperatures and transfer frozen vitreous samples as demonstrated by vibrational spectroscopy. Our findings demonstrate that the cryo-transfer station performs comparably to existing commercial models, while offering greater accessibility and customizability. The design for the station is open source to encourage other groups to replicate and build on this development. We hope that this project will increase access to cryo-transfer stations for researchers in a variety of disciplines with nonstandard equipment.

Assessing the risk of climate maladaptation for Canadian polar bears.

The Arctic is warming four times faster than the rest of the world, threatening the persistence of many Arctic species. It is uncertain if Arctic wildlife will have sufficient time to adapt to such rapidly warming environments. We used genetic forecasting to measure the risk of maladaptation to warming temperatures and sea ice loss in polar bears (Ursus maritimus) sampled across the Canadian Arctic. We found evidence for local adaptation to sea ice conditions and temperature. Forecasting of genome-environment mismatches for predicted climate scenarios suggested that polar bears in the Canadian high Arctic had the greatest risk of becoming maladapted to climate warming. While Canadian high Arctic bears may be the most likely to become maladapted, all polar bears face potentially negative outcomes to climate change. Given the importance of the sea ice habitat to polar bears, we expect that maladaptation to future warming is already widespread across Canada.

Accumulation of Environmental Radioactivity on the Surface of a High Arctic Ice Cap (Flade Isblink, NE Greenland).

Under climatic warming, glaciers are becoming a secondary source of atmospheric contaminants originally released into the environment decades ago. This phenomenon has been well-documented for glaciers near emission sources. However, less is known about polar ice sheets and ice caps. Radionuclides are one of the contaminants that can be remobilised through ice melting and accumulate in cryoconite material on the surface of glaciers. To understand the cycling of radionuclides in polar glacial contexts, we evaluate the radioactivity of cryoconite samples from Flade Isblink, a High Arctic ice cap in northeast Greenland. The measured radioactivity is among the highest reported across the High Arctic and the highest from Greenland. The high variability observed among the samples is explained by considering the different macroscopic features of single cryoconite deposits. The radioactivity source is compatible with the stratospheric reservoir established during atmospheric nuclear tests and with weapons-grade fissile fuel, likely originating from Novaya Zemlya proving grounds. This study shows that the ability of cryoconite to accumulate radioactivity in remote areas is undisputed, highlighting the need for a deeper understanding of the remobilisation of radioactive species in polar glacial contexts.

Hypofractionated radiotherapy combined with bevacizumab plus low-dose ifosfamide, carboplatin, and etoposide as second-line chemoradiotherapy for progressing spinal diffuse midline glioma, H3K27-altered: illustrative case.

BACKGROUND: Spinal cord diffuse midline glioma (DMG) is a relatively rare disease with a poor prognosis and no effective treatment. OBSERVATIONS: A 45-year-old man presented with rapidly progressive paraplegia in both lower extremities, along with bladder and bowel disturbance. Spinal magnetic resonance imaging (MRI) showed a heterogeneously contrast-enhanced mass at the T1-4 levels. A biopsy via T1-4 decompressive laminectomy with expansive duraplasty was performed. The histopathological diagnosis was DMG, H3K27-altered, World Health Organization grade 4. Radiation plus concomitant temozolomide was started; however, follow-up MRI showed tumor progression. Additional hypofractionated radiotherapy (HFRT; 24 Gy/5 fractions) was performed, with bevacizumab (BEV) plus low-dose ifosfamide-carboplatin-etoposide (ICE) as second-line treatment. One month later, MRI showed tumor regression with significant improvement in the peritumoral edema. The chemotherapy regimen was repeated every 4-6 weeks, and the patient remained stable. After 13 courses of chemotherapy, the size of the spinal DMG increased markedly, with dissemination to the temporal lobe. The patient died approximately 21 months after the initial diagnosis. LESSONS: Spinal DMG is a malignant tumor with a poor prognosis. However, treatment with additional HFRT combined with BEV plus low-dose ICE may inhibit tumor progression to prolong the progression-free period and survival. https://thejns.org/doi/10.3171/CASE2464.

Ice Nucleation Mechanisms on Platinum Surfaces in PEM Fuel Cells: Effects of Surface Morphology and Wettability.

Understanding the ice nucleation mechanism in the catalyst layers (CLs) of proton exchange membrane (PEM) fuel cells and inhibiting icing by designing the CLs can optimize the cold start strategies, which can enhance the performance of PEM fuel cells. Herein, mitigating the structural matching and templating effects by adjusting the surface morphology and wettability can inhibit icing on the platinum (Pt) catalyst surface effectively. The Pt(211) surface can inhibit icing because the atomic spacing of (211) crystalline surface is much larger than the characteristic distance of ice crystal, thereby mitigating the structural matching effects. A water overlayer on the Pt surface induced by the strong attraction of Pt can act as a template for ice layers and plays an important role in the icing process. Buckling of water overlayer due to the larger atomic spacing of (211) crystalline surface mitigates the templating effect and inhibits icing. Moreover, the water overlayer on the hydrophobic Pt(211) surface with fewer water molecules also mitigates the templating effect, which makes ice nucleation more difficult than homogeneous nucleation. These findings reveal the ice nucleation mechanisms on the Pt catalyst surface from the molecular level and are valuable for catalyst designs to inhibit icing in CL.

The effect of a combined cooling intervention on cognitive function in the heat during an intermittent running protocol.

Despite optimal cognitive function being essential for performance, there is a lack of research on the effectiveness of combined cooling interventions on team sport athlete's cognitive function when exercising in the heat. In a randomised, crossover design, 12 unacclimatised men (age: 22.3 ± 3.0 years, body mass: 73.4 ± 5.1 kg, height: 181.0 ± 5.3 cm and V ˙ O 2 $\dot{\mathrm{V}}{\mathrm{O}}_{2}$ max: 51.2 ± 9.5 mL/kg/min) participated in a control (CON) and combined cooling trial (ice slurry and ice collar; COOL). A battery of cognitive tests were completed prior to, during (at half-time) and following a 90-min intermittent running protocol in the heat (33°C, 50% relative humidity (RH)). Perceptual and physiological measures were taken throughout the protocol. In CON, response times were quicker on the Stroop task complex level (p = 0.002) and the visual search test complex level at full-time (p = 0.014) compared to COOL. During COOL, response times were quicker at half-time on the Stroop task complex level (p = 0.024) compared to CON. Lower rectal temperatures were seen during COOL (CON: 37.44 ± 0.65°C and COOL: 37.28 ± 0.68°C) as well as lower skin, neck and forehead temperatures (main effect of trial, all p < 0.05). Lower ratings of thermal sensation and perceived exertion and enhanced thermal comfort were recorded during COOL (main effect of trial, all p < 0.05). Whilst minimal differences in cognitive function were found when using the combined cooling intervention, the findings highlight a practical and effective strategy to improving many physiological and perceptual responses to intermittent exercise in the heat.

Sustainable Manufacturing of Multifunctional Fluorine-Free Superslippery Flexible Surfaces.

Surface contamination and friction result in significant energy losses with widespread environmental impact. In the present work, we developed fluorine-free super-slippery surfaces employing environmentally friendly and simple biofuel-based flame treatment of polydimethylsiloxane (PDMS). Through a unique combination of processing parameters, highly transparent (>90%) and flexible films were engineered with omniphobic, anti-icing, and ultra-low friction properties. The processed films showed an extremely low tilting angle (<5°) and contact angle hysteresis (<4°) against different liquids, even under subzero temperatures. The coefficient of friction (COF) reduced to 0.01 after processing compared to ∼1 for PDMS. Extremely low ice adhesion of <20 kPa and enhanced freezing time ensured anti-icing behavior. The exceptional multidimensional traits were derived from the extremely stable silicone lubricant layer ensured by the hierarchically structured wrinkles. Wind tunnel tests showed that an air drag velocity of less than 0.5 m/s was sufficient to initiate droplet motion, highlighting low interfacial friction that leads to an anti-staining nature. Sustaining the self-cleaning and anti-staining characteristics, the processed surface showed utmost durability under different harsh conditions. The super-slippery surfaces with multifunctional characteristics fabricated through a sustainable route can be effectively used for various engineering and industrial applications.

Relationships between sprint skating performance and insole plantar forces in national-level hockey athletes.

BACKGROUND: Sprint skating is essential for competitive success in hockey. Previous studies have highlighted various measures of lower-body strength and power as key factors influencing sprint performance. However, while these studies have indicated a significant association between the ability to exert greater force and impulse into the ice surface, and the capacity to achieve faster sprint skating speeds, the direct relationship between these factors remains largely inferred. RESEARCH QUESTION: What are the relationships between insole plantar force variables, sprint skating performance, and their association with physical capacity measures, in national-level male hockey athletes? METHODS: Athletes (n=13) performed 25 m sprint skating trials with insole force sensors and completed: ankle dorsiflexion and hip abduction range-of-motion (ROM), countermovement jump (CMJ), seated single-leg jump, and 10-5 repeated-hop test (RHT) assessments. RESULTS: Relationships were assessed using Kendall's Tau rank correlations (τ), with significant relationships identified between mean relative weight acceptance impulse [WAI] and 0-5 m (τ=0.47) and total distance (τ=0.46) times. Additionally, significant associations were observed between mean relative WAI and: CMJ relative eccentric deceleration impulse (τ=0.44), CMJ eccentric peak velocity (τ=-0.46) and RHT concentric impulse (τ=-0.56). Finally, significant relationships were identified between mean relative PI for all strides and the 10-20 m split, with peak velocity (PV) (τ=-0.58 to -0.73); and between ankle dorsiflexion ROM and PV (τ=-0.57). SIGNIFICANCE: Athletes with faster initial acceleration and overall sprint performance times demonstrated lower relative WAI during their strides and employed a jump strategy that optimized concentric impulse with a rapid eccentric phase. To attain a high PV the athletes appeared to require a stride that maximized glide and minimized vertical force, with greater ankle dorsiflexion ROM potentially facilitating this.

Ice formation and its elimination in cryopreservation of oocytes.

Damage from ice and potential toxicity of ice-inhibiting cryoprotective agents (CPAs) are key issues in assisted reproduction of humans, domestic and research animals, and endangered species using cryopreserved oocytes and embryos. The nature of ice formed in bovine oocytes (similar in size to oocytes of humans and most other mammals) after rapid cooling and during rapid warming was examined using synchrotron-based time-resolved x-ray diffraction. Using cooling rates, warming rates and CPA concentrations of current practice, oocytes show no ice after cooling but always develop large ice fractions-consistent with crystallization of most free water-during warming, so most ice-related damage must occur during warming. The detailed behavior of ice at warming depended on the nature of ice formed during cooling. Increasing cooling rates allows oocytes soaked as in current practice to remain essentially ice free during both cooling and warming. Much larger convective warming rates are demonstrated and will allow routine ice-free cryopreservation with smaller CPA concentrations. These results clarify the roles of cooling, warming, and CPA concentration in generating ice in oocytes and establish the structure and grain size of ice formed. Ice formation can be eliminated as a factor affecting post-warming oocyte viability and development in many species, improving outcomes and allowing other deleterious effects of the cryopreservation cycle to be independently studied.

Identification of novel Tet(X6)-Tet(X2) recombinant variant in Elizabethkingia meningoseptica from a bullfrog farm and downstream river in China.

Introduction: The dissemination of strains producing tetracyclines monooxygenase Tet(X) from breeding farms to the natural environment poses a potential threat to public health. Methods: Antimicrobial susceptibility testing and WGS were performed to identify resistance phenotypes and genotypes. Cloning experiments, sequence alignment, and homology modeling were used to characterize the function and formation mechanisms of the recombinant variant. The mobilization potential of Tet(X) was assessed by collinearity analysis, conjugation experiments, and phylogenetic analysis. Results: Three tet(X)-producing Elizabethkingia meningoseptica strains were isolated from bullfrog breeding ponds, the sewage outlet, and downstream river in Zhejiang Province, China. These strains carry a novel Tet(X) variant, differing from Tet(X6) by seven residues, and possess the ability to degrade tetracyclines. Interestingly, the novel Tet(X) is a recombinant variant formed by homologous recombination of Tet(X6) and the C-terminal of Tet(X2). Further analysis revealed that Tet(X6) formed several Tet(X) variants, including Tet(X5), through homologous recombination. The novel tet(X) gene is located on a circularizable integrative and conjugative element (ICEEmeChn3), with ISwz1 participating in the recombination of its multi-drug resistance region, potentially facilitating the mobilization and recombination of tet(X) in early hosts. These three strains were clonally transmitted and shared a close genetic relationship (SNP < 62) with a clinically-sourced strain isolated from the same province. Discussion: To our knowledge, this is the first report of homologous recombination between Tet(X) variants with differing activities. These clonal strains provide evidence of the transmission of tet(X)-positive strains from aquaculture sewage to the natural environment, highlighting the need to strengthen the monitoring and management of this emerging farming model.

Characterizing offshore polar ocean soundscapes using ecoacoustic intensity and diversity metrics.

Polar offshore environments are considered the last pristine soundscapes, but accelerating climate change and increasing human activity threaten their integrity. In order to assess the acoustic state of polar oceans, there is the need to investigate their soundscape characteristics more holistically. We apply a set of 14 ecoacoustic metrics (EAMs) to identify which metrics are best suited to reflect the characteristics of disturbed and naturally intact polar offshore soundscapes. We used two soundscape datasets: (i) the Arctic eastern Fram Strait (FS), which is already impacted by anthropogenic noise, and (ii) the quasi-pristine Antarctic Weddell Sea (WS). Our results show that EAMs when applied in concert can be used to quantitatively assess soundscape variability, enabling the appraisal of marine soundscapes over broad spatiotemporal scales. The tested set of EAMs was able to show that the eastern FS, which is virtually free from sea ice, lacks seasonal soundscape dynamics and exhibits low acoustic complexity owing to year-round wind-mediated sounds and anthropogenic noise. By contrast, the WS exhibits pronounced seasonal soundscape dynamics with greater soundscape heterogeneity driven in large part by the vocal activity of marine mammal communities, whose composition in turn varies with the prevailing seasonal sea ice conditions.

SPOT-RASTR-A cryo-EM specimen preparation technique that overcomes problems with preferred orientation and the air/water interface.

In cryogenic electron microscopy (cryo-EM), specimen preparation remains a bottleneck despite recent advancements. Classical plunge freezing methods often result in issues like aggregation and preferred orientations at the air/water interface. Many alternative methods have been proposed, but there remains a lack a universal solution, and multiple techniques are often required for challenging samples. Here, we demonstrate the use of lipid nanotubes with nickel NTA headgroups as a platform for cryo-EM sample preparation. His-tagged specimens of interest are added to the tubules, and they can be frozen by conventional plunge freezing. We show that the nanotubes protect samples from the air/water interface and promote a wider range of orientations. The reconstruction of average subtracted tubular regions (RASTR) method allows for the removal of the nanotubule signal from the cryo-EM images resulting in isolated images of specimens of interest. Testing with ß-galactosidase validates the method's ability to capture particles at lower concentrations, overcome preferred orientations, and achieve near-atomic resolution reconstructions. Since the nanotubules can be identified and targeted automatically at low magnification, the method enables fully automated data collection. Furthermore, the particles on the tubes can be automatically identified and centered using 2D classification enabling particle picking without requiring prior information. Altogether, our approach that we call specimen preparation on a tube RASTR holds promise for overcoming air-water interface and preferred orientation challenges and offers the potential for fully automated cryo-EM data collection and structure determination.

In-situ aircraft observations of aerosol and cloud microphysical characteristics of mixed-phase clouds over the North China Plain.

Aerosol-cloud interactions play a vital role in climate change. This study leverages observations from the King-350 aircraft over the North China Plain on November 29, 2019, to examine aerosol and cloud microphysical characteristics of mixed-phase clouds. Through detailed vertical and spectral distributions, we investigate aerosol, cloud droplet, and ice crystal distributions in seeded clouds (SC) and natural precipitating clouds (NPC) within the same cloud system. From the vertical profile, SC and NPC have similar vertical distributions of aerosol and cloud droplets, with over 95 % of aerosols concentrated below 1600 m near the ground. Cloud droplets are more evenly distributed within the two clouds, cloud droplet number concentrations (Nc) in SC were three orders of magnitude higher than in NPC. Ice water content (IWC) and ice crystal number concentration (Ni) show distinct layer preferences-accumulating predominantly in SC's top layer and NPC's middle layer. From spectral distribution, a smaller proportion of cloud droplets (40-50 µm in diameter, the same hereafter) in SC compared to NPC. Rimed ice crystals and globular graupel (1325-1550 µm in diameter) were in SC, while plate and irregular ice crystals (300-450 µm) were in NPC with an order of magnitude higher than in SC. These microphysical differences highlight the complexity of cloud seeding efficacy, which varies based on cloud conditions and microphysical properties. In the first seeding, Ni increased by 1-2 orders of magnitude (125-300 µm) in the high Nc (Nc > 1.11 × 105 L-1) region. Seeding in low Nc (Nc < 1.11 × 105 L-1) regions was hard to be effective, especially in low Nc and low liquid water content (LWC) (LWC < 0.122 g/m3) regions. In the second seeding, ice crystals (125-250 µm) produced by the first seeding enhance the seeding efficiency. The responded regions were more sensitive to subsequent seeding, resulting in stronger reactions or longer duration.