Sensitive and specific detection of trace Al3+ ions using an upconversion nanoparticle-xylenol orange complex via the inner filter effect.

We have developed a novel fluorescence sensor based on upconversion nanoparticles (UCNPs) for the rapid and sensitive detection of trace aluminum ions (Al3+). The sensor utilizes the inner filter effect (IFE) between the UCNPs and the xylenol orange-aluminum complex (XO-Al3+), resulting in significant fluorescence quenching at 543 nm upon Al3+ binding. This quenching correlates directly with the Al3+ concentration, allowing for quantitative detection within a range of 0-30 µM and achieving an ultra-low detection limit of 0.19 µM. Selectivity of the sensor is enhanced by the incorporation of ascorbic acid, which masks interfering Fe3+ ions, thus ensuring accurate determination of Al3+ even in the presence of other metal ions. The UCNPs-XO sensor exhibits excellent stability and reproducibility, and minimal interference from commonly co-existing substances. This makes it suitable for the detection of Al3+ in various matrices, including food products and environmental water samples. Our work offers a significant advancement in Al3+ detection, with potential applications in food safety, environmental monitoring, and public health.

Wool powder assisted colorimetric sensing yarn with high sensitivity for NH3 monitoring.

Colorimetric sensors have applications in gas monitoring due to their simple and quick detection through visible color changes. However, it remains challenging to prepare colorimetric sensors with high sensitivity. Herein, this work fabricated a biomass-based colorimetric sensing yarn with high sensitivity using anthocyanins as the colorimetric dye and wool powder as an effective ammonia (NH3) adsorbent. The sensitivity of the prepared yarns was evaluated for detection limit and response time. Surprisingly, the addition of 3% wool powder greatly improved the sensitivity of the prepared yarns, with a reduction of both detection limit and responsive time from 100 ppm to 20 ppm, and 2 min to 20 s, respectively when exposed in 150 ppm NH3. The prepared yarns also showed good selectivity and reusability. An example of the practical use of colorimetric yarns was presented. This work provides a facile strategy for fabricating wearable devices for toxic gas monitoring with visual output.

Enterovibrio qingdaonensis sp. nov. and Enterovibrio gelatinilyticus sp. nov., two marine bacteria isolated from surface seawater of Qingdao offshore.

Two Gram-stain-negative, facultatively anaerobic, motile, and rod-shaped marine bacteria, designated strains ZSDZ35T and ZSDZ42T, were isolated from surface seawater of Qingdao offshore. Phylogenetic analysis of the 16S rRNA genes and whole genome data placed ZSDZ35T and ZSDZ42T within the genus Enterovibrio. Strain ZSDZ35T was most closely related to Enterovibrio nigricans DSM 22720T with 97.55% sequence similarity, whereas ZSDZ42T was most closely related to Enterovibrio calviensis DSM 14347T with 98.97% sequence similarity. Strain ZSDZ35T grew with 0-8% (w/v) NaCl (optimum 4%), at 16-37 °C (optimum 28 °C) and pH 6.0-9.0 (optimum pH 8.0); whereas strain ZSDZ42T grew with 1-7% (w/v) NaCl (5%), at 4-28 °C (8 °C) and pH 6.0-9.0 (pH 7.0). Both strains shared the same major fatty acid components of summed feature 3 (C16:1ω7c or/and C16:1ω6c), summed feature 8 (C18:1ω7c and C18:1ω6c) and C16:0, with different proportions. The DNA G + C contents of strains ZSDZ35T and ZSDZ42T were 47.2% and 46.7%, respectively. Based on the results of polyphasic analyses, ZSDZ35T and ZSDZ42T are considered to represent novel species, for which the names Enterovibrio qingdaonensis sp. nov. (type strain, ZSDZ35T = MCCC 1K06293T = KCTC 82887T) and Enterovibrio gelatinilyticus sp. nov. (type strain, ZSDZ42T = MCCC 1K06294T = KCTC 82886T) are proposed, respectively.

Air pollution's numerical, spatial, and temporal heterogeneous impacts on childhood hand, foot and mouth disease: a multi-model county-level study from China.

BACKGROUND: While stationary links between childhood hand, foot and mouth disease (HFMD) and air pollution are known, a comprehensive study on their heterogeneous relationships (nonstationarity), jointly considering numerical, temporal and spatial dimensions, has not been reported. METHODS: Monthly HFMD incidence and air pollution data were collected at the county level from Sichuan-Chongqing, China (2009-2011), alongside meteorological and social environmental covariates. Key influential factors were identified using random forest (RF) under the stationary assumption. Factors' numerically, temporally, and spatially heterogeneous relationships with HFMD were assessed using generalized additive model (GAM) and geographically and temporally weighted regression (GTWR). RESULTS: Our findings highlighted the relatively higher stationary contributions of fine particulate matter (PM2.5) and ozone (O3) to HFMD incidence across Sichuan-Chongqing counties. We further uncovered heterogeneous impacts of PM2.5 and O3 from three nonstationary perspectives. Numerically, PM2.5 showed an inverse 'V'-shaped relationship with HFMD incidence, while O3 exhibited a complex pattern, with increased HFMD incidence at low PM2.5 and moderate O3 concentrations. Temporally, PM2.5's impact peaked in autumn and was weakest in spring, whereas O3's effect was strongest in summer. Spatially, hotspot mapping revealed high-risk clusters for PM2.5 impact across all seasons, with notable geographical variations, and for O3 in spring, summer, and autumn, concentrated in specific regions of Sichuan-Chongqing. CONCLUSIONS: This study underscores the nuanced and three-perspective heterogeneous influences of air pollution on HFMD in small areas, emphasizing the need for differentiated, localized, and time-sensitive prevention and control strategies to enhance the precision of dynamic early warnings and predictive models for HFMD and other infectious diseases, particularly in the fields of environmental and spatial epidemiology.

Genetically Encoded Fluorescence Resonance Energy Transfer Biosensor for Live-Cell Visualization of Lamin A Phosphorylation at Serine 22.

Extensive phosphorylation at serine 22 (pSer22) on lamin A is the hallmark of cell mitosis, which contributes to the breakdown of nuclear envelope. In the interphase, pSer22 lamin A exists in low abundance and is involved in mechanotransduction, virus infection, and gene expression. Numerous evidences emerge to support lamin A regulation on cell function and fate by phosphorylation. However, live-cell imaging tools for visualizing the dynamics of pSer22 lamin A are yet to be established. Herein, we developed a novel lamin A phosphorylation sensor (LAPS) based on fluorescence resonance energy transfer (FRET) with high sensitivity and specificity. We observed the dynamic lamin A phosphorylation during the cell cycle progression in single living cells: the increase of pSer22 modification when cells entered the mitosis and recovered upon the mitosis exit. Our biosensor also showed the gradual reduction of pSer22 modification during cell adhesion and in response to hypotonic environment. By applying LAPS, we captured the propagation of pSer22 modification from inside to outside of the inner nuclear membrane, which further led to the breakdown of nuclear envelope. Meanwhile, we found the synchronous phosphorylation of pSer22 lamin A and H3S10ph at mitosis entry. Inhibition of Aurora B, the responsible kinase for H3S10ph, could shorten the mitotic period without obvious effect on the pSer22 modification level of lamin A. Thus, LAPS allows the spatiotemporal visualization of the lamin A pSer22, which will be useful for elucidating the molecular mechanisms underlying cell mitosis and mechanoresponsive processes.

Exosomal PDL1 Suppresses the Anticancer Activity of CD8+ T Cells in Hepatocellular Carcinoma.

Tumor microenvironment (TME) is essential for the development and progression of hepatocellular carcinoma (HCC). Exosomes participate in constructing TME by passing biological information, but the regulatory effect of PDL1 in exosomes on anticancer activity of CD8+ T cells in HCC still needs to be further explored. In this study, high level of PDL1 was found in plasma exosomes of HCC patients, which turned out to be significantly associated with the increased number of tumor nodules, the upregulated level of serum AFP, the raised tendency of TNM stage, and the poor prognosis of HCC. The expression of CD8 may be inhibited in HCC that is characterized with high level of PDL1, and the protein level of exosomal PDL1 was determined by intracellular PDL1 abundance. High level of exosomal PDL1 inhibited the proliferation and activation of CD8+ T cells, but exhibited limited effect on the proliferation of hepatic cancer cells. Moreover, the growth of tumors formed by hepatic cancer cells Hepa1-6 in C57L mice was significantly promoted by the exosomal PDL1, which might be caused by the inhibitory effect of exosomal PDL1 on CD8+ T cells. Thus, exosomal PDL1 promotes the development and progression of HCC through inhibiting the anticancer activity of CD8+ T cells. This study provides sights for understanding the oncogenic role of PDL1 and a reasonable explanation for the low efficacy of anti-PD1/PDL1 immunotherapies in HCC.

Pneumocystis pneumonia in stage IIIA lung adenocarcinoma with immune-related acute kidney injury and thoracic radiotherapy: A case report.

BACKGROUND: Immune checkpoint inhibitors (ICIs) are therapeutic agents for advanced and metastatic non-small cell lung cancer (NSCLC) with high clinical antitumor efficacy. However, immune-related adverse events occur in 20% of these patients and often requiring treatment with immunosuppressive agents, such as corticosteroids. Consequently, this may increase the risk of patients to opportunistic infections. Pneumocystis jirovecii pneumonia (PJP), a rare but serious opportunistic infection typically observed in patients with human immunodeficiency virus, can also occur in cancer patients undergoing long-term glucocorticoid treatment. CASE SUMMARY: We report a case of a 56-year-old male with squamous NSCLC treated with triplimab combined with paclitaxel, carboplatin, and radical thoracic radiation therapy. Following this regimen, he developed acute kidney injury (AKI) with elevated creatinine levels. After concurrent radical chemoradiotherapy ended, he developed a grade 3 immune-related AKI. High-dose corticosteroids were administered to treat AKI, and renal function gradually recovered. Corticosteroids were reduced to a dose of 10 mg prednisone equivalent daily eight weeks later; however, he developed severe pneumonia with spontaneous pneumothorax. Next-generation sequencing of the bronchoscopic lavage revealed PJP co-infection with herpes simplex virus 1 and cytomegalovirus. The inflammation was more severe in areas exposed to radiation. Piperacillin-tazobactam, acyclovir, sulfamethoxazole, and trimethoprim were used to control the infection. The patient recovered, and immunotherapy was terminated. CONCLUSION: PJP is rare but can occur in patients with ICI adverse events and should be differentiated from tumor progression or immune-related adverse events. Thoracic radiation may increase risk, necessitating careful monitoring and prevention.

Effects of orthogonal dual-frequency ultrasound-assisted treatment combined with bioactive coating containing Melissa officinalis L. essential oil on changes in quality, lipid, and protein of large yellow croaker (Pseudosciaena crocea) during cold storage.

How to reduce the quality loss of aquatic products during storage is a topic worth exploring. This study proposed a method combining orthogonal dual-frequency ultrasound-assisted treatment (20 kHz vertically, 40 kHz horizontally, 400 W) with bioactive coating (Melissa officinalis L. essential oil-carboxymethyl chitosan-locust bean gum) and discussed the effects of this combined treatment on the quality, lipid, and protein of large yellow croaker during cold storage (4 °C). The results showed that both ultrasound-assisted treatment (US) and bioactive coating (CMCS) significantly inhibited microbial growth and quality deterioration in the fish, with the combined treatment group (US+CMCS) showing the best effect. The shelf life of large yellow croaker in the control group (CK) was 6 d, while the shelf life for US, CMCS, and US+CMCS treatments was 12 d, 12 d, and 18 d, respectively. Additionally, the combined treatment inhibited lipid oxidation and effectively delayed the oxidative degradation of protein in the large yellow croaker during cold storage. Therefore, the method of orthogonal dual-frequency ultrasound-assisted treatment (20 kHz vertically, 40 kHz horizontally, 400 W) combined with bioactive coating (Melissa officinalis L. essential oil-carboxymethyl chitosan-locust bean gum) proposed in this study was a promising approach for the preservation of aquatic products during storage.

Scaling of cotyledon and primary leaf mass versus area in Acer platanoides seedlings under different light conditions.

Cotyledons play an important role in early seedling establishment. However, relative to primary leaves, cotyledons tend to have a different investment-on-return strategy. To detect the potential differences in the mass (M) versus area (A) scaling relationships between cotyledons and primary leaves in different light environments, a total of 75 Acer platanoides seedlings were sampled at an open site (n = 52; light availability: 74 ±â€…5 %) and a shaded site (n = 23; light availability: 4.2 ±â€…1.2 %). Reduced major axis regression protocols were used to fit the M versus A scaling relationships of primary leaves and cotyledons. The bootstrap percentile method was used to test the significance of the differences in the scaling exponents of M versus A between the two light environments. The scaling exponents of cotyledons at both two sites, as well as the primary leaves at the shade site, were greater than unity indicating 'diminishing returns', while the scaling exponent of primary leaves at the open site was smaller than unity indicating 'increasing returns'. The data collectively indicated light-dependent shifts in support investments and differences in the function of cotyledons and primary leaves. Average leaf structural traits displayed significant differences between the two light environments in accordance with the premium in enhancing photosynthetic capacity in high light and light interception in low light. Although the trait responses to light availability were similar for primary leaves and cotyledons, primary leaves were more responsive to light availability, indicating lower plasticity of cotyledons in response to light levels. These results advance our understanding of the roles of cotyledons and primary leaves in the life history of seedlings in different forest light environments.

Bacterial dynamics in the progression of caries to apical periodontitis in primary teeth of children with severe early childhood caries.

Background: Early childhood caries (ECC) are a prevalent chronic disease in young children. However, there has been limited research on the microbiota in different tissue levels of the same tooth in children with ECC. This study aimed to investigate the dynamic changes in bacterial diversity during the progression of Severe Early Childhood Caries (S-ECC) within the same tooth, from the tooth surface to the root canal, by collecting tissue samples from different areas of the affected tooth. Methods: Twenty primary teeth with periapical periodontitis were selected from 20 children aged 3-5 years, with 100 samples collected from the different layers: uncavitated buccal enamel surface without white spot lesion (surface), the outermost layer of the dentin carious lesion (superficial), the inner layer of carious dentin (deep), necrotic pulp tissue (pulp), and root exudate (exudate). The taxonomy of each OTU representative sequence was analyzed against the 16S rRNA database. Comparisons of alpha diversity between groups were performed. The number of shared and unique genera between groups counted. Beta diversity was contrasted to evaluate differences in bacterial community composition, and the relationships between the microbiota and samples were analyzed. The heatmap analysis of the 30 most abundant genera was used, which highlighted their relative distribution and abundance. The significantly abundant taxa (phylum to genera) of bacteria among the different groups were identified. The differences of relative abundance between bacterial genera among the five groups were analyzed. Significant Spearman correlations were noted, and visualization of the co-occurrence network was conducted. Results: Bacterial 16S rRNA gene sequencing showed that most genera were present in all layers, with the number of shared genera increasing as the disease advanced. The bacterial communities and core genera in the co-occurrence network changed with progression to severe ECC. Conclusion: An increase in both the quantity and complexity of bacterial interactions was observed. This study emphasized the importance of paying attention to the relationship between microbial species rather than just checking changes in bacterial species structure when investigating the role of bacteria in disease progression.

Evolution and driving mechanism of multiple ecosystem services in resource-based region of Northern China.

Understanding the spatiotemporal characteristics and comprehensive service capabilities of various ecosystem services is crucial for maintaining regional ecosystem security, and clarifying the driving mechanisms of ecosystem services plays a guarantee for achieving regional sustainable development. Based on the ecological issues of Shanxi Province (SXP) in China, an assessment system covering eight targeted ecosystem services were constructed to quantitatively analyze the spatio-temporal patterns and contribution rates of driving factors. The water conservation, sand fixation, environment purification and habitat quality in the Loess Hills of western SXP have improved, with significant increase in hotspots. The comprehensive service capacity of ecosystem services in the Fenwei Basin (central SXP) has deteriorated, and the coldspots have expanded. The water conservation, carbon storage, habitat quality and recreation culture in the Yanshan-Taihang Mountains (eastern SXP) were enhanced, while product supply, carbon storage and environment purification were deteriorated. Land use is the dominant influencing factor on product supply. Water and soil conservation, sand fixation, and environment purification are dominated by climate factors. Carbon storage, habitat quality and recreation culture are influenced by underlying surface conditions. The current study provided a research paradigm, which will help the government with appropriate management policies to ensure the effectiveness of ecological protection and restoration, and offers insights for facilitating ecological sustainability and economic transformation in resource-based regions worldwide.

Corrigendum: Tribological properties of MAO ceramic coatings with annulus array texture on disposable surgical gloves.

[This corrects the article DOI: 10.3389/fbioe.2024.1397050.].

Responsive circularly polarized ultralong room temperature phosphorescence materials with easy-to-scale and chiral-sensing performance.

Circularly polarized room temperature phosphorescence materials represent a state-of-the-art frontier of optical materials and exhibit promising applications in various fields. Herein, we fabricate a series of full-color circularly polarized room temperature phosphorescence materials, based on anionic cellulose derivatives and achiral luminophores. The ionic achiral substituents promote the spontaneous formation of chiral helical structure of cellulose derivatives via the electrostatic repulsion effect. There are multiple interactions between anionic cellulose derivatives and the doped luminophores, thus the chirality is transferred to luminophores and the non-radiative transition is inhibited. The resultant materials can be easily processed into large-scale film and flexible 3D objects with repeatable folding and curling properties. In addition, their phosphorescence performance shows to be excitation-dependence, time-dependence, visible-light excitation, and multi-responsiveness to humidity, temperature as well as pH value. Importantly, they recognize many enantiomers in an instrument-free visual mode, including amino acids, hydroxyl acids, organic phosphate and hydrobenzoin. These results provide insights into design of advanced optical materials which can be applied in multilevel information handling and chiral sensing.

Modeling tuberculosis transmission flow in China, 2010-2012.

BACKGROUND: China has the third largest number of TB cases in the world, and the average annual floating population in China is more than 200 million, the increasing floating population across regions has a tremendous potential for spreading infectious diseases, however, the role of increasing massive floating population in tuberculosis transmission is yet unclear in China. METHODS: 29,667 tuberculosis flow data were derived from the new smear-positive pulmonary tuberculosis cases in China. Spatial variation of TB transmission was measured by geodetector q-statistic and spatial interaction model was used to model the tuberculosis flow and the regional socioeconomic factors. RESULTS: Tuberculosis transmission flow presented spatial heterogeneity. The Pearl River Delta in southern China and the Yangtze River Delta along China's east coast presented as the largest destination and concentration areas of tuberculosis inflows. Socioeconomic factors were determinants of tuberculosis flow. Some impact factors showed different spatial associations with tuberculosis transmission flow. A 10% increase in per capita GDP was associated with 10.2% in 2010 or 2.1% in 2012 decrease in tuberculosis outflows from the provinces of origin, and 1.2% in 2010 or 0.5% increase in tuberculosis inflows to the destinations and 18.9% increase in intraprovincial flow in 2012. Per capita net income of rural households and per capita disposable income of urban households were positively associated with tuberculosis flows. A 10% increase in per capita net income corresponded to 14.0% in 2010 or 3.6% in 2012 increase in outflows from the origin, 44.2% in 2010 or 12.8% increase in inflows to the destinations and 47.9% increase in intraprovincial flows in 2012. Tuberculosis incidence had positive impacts on tuberculosis flows. A 10% increase in the number of tuberculosis cases corresponded to 2.2% in 2010 or 1.1% in 2012 increase in tuberculosis inflows to the destinations, 5.2% in 2010 or 2.0% in 2012 increase in outflows from the origins, 11.5% in 2010 or 2.2% in 2012 increase in intraprovincial flows. CONCLUSIONS: Tuberculosis flows had clear spatial stratified heterogeneity and spatial autocorrelation, regional socio-economic characteristics had diverse and statistically significant effects on tuberculosis flows in the origin and destination, and income factor played an important role among the determinants.

Toward industry 5.0: Challenges and enablers of intelligent manufacturing technology implementation under the perspective of sustainability.

The advancement of intelligent manufacturing technology in the era of Industry 5.0 has propelled the intelligence and automation of manufacturing production, while also exerting a significant impact on sustainable development of the manufacturing industry. However, the challenges and enablers faced by the transformation of intelligent manufacturing technology in the context of sustainable development of Industry 5.0 are still unclear. Based on literature review and expert opinions, this study uses the Likert scale to determine the challenges and enablers of the implementation of intelligent manufacturing technology in social, environmental and economic sustainability. The fuzzy-DEMETAL and AISM are used to analyze the logical relationship and hierarchical relationship between the above factors, and the MICMAC matrix is used to determine the key influencing factors. The research conclusions show that the most important challenges affecting the implementation of intelligent manufacturing technology are cost and funding, and the most important enabler is social benefits and public service improved. This research will provide insights for industry practitioners and decision makers in the management and decision-making process of implementing the transformation and upgrading of manufacturing intelligent manufacturing, thereby enhancing the sustainability of manufacturing development.

DualStreamFoveaNet: A Dual Stream Fusion Architecture with Anatomical Awareness for Robust Fovea Localization.

Accurate fovea localization is essential for analyzing retinal diseases to prevent irreversible vision loss. While current deep learning-based methods outperform traditional ones, they still face challenges such as the lack of local anatomical landmarks around the fovea, the inability to robustly handle diseased retinal images, and the variations in image conditions. In this paper, we propose a novel transformer-based architecture called DualStreamFoveaNet (DSFN) for multi-cue fusion. This architecture explicitly incorporates long-range connections and global features using retina and vessel distributions for robust fovea localization. We introduce a spatial attention mechanism in the dual-stream encoder to extract and fuse self-learned anatomical information, focusing more on features distributed along blood vessels and significantly reducing computational costs by decreasing token numbers. Our extensive experiments show that the proposed architecture achieves state-of-the-art performance on two public datasets and one large-scale private dataset. Furthermore, we demonstrate that the DSFN is more robust on both normal and diseased retina images and has better generalization capacity in cross-dataset experiments.

Climate factors dominate the elevational variation in grassland plant resource utilization strategies.

Specific leaf area (SLA) and leaf dry matter content (LDMC) are key leaf functional traits often used to reflect plant resource utilization strategies and predict plant responses to environmental changes. In general, grassland plants at different elevations exhibit varying survival strategies. However, it remains unclear how grassland plants adapt to changes in elevation and their driving factors. To address this issue, we utilized SLA and LDMC data of grassland plants from 223 study sites at different elevations in China, along with climate and soil data, to investigate variations in resource utilization strategies of grassland plants along different elevational gradients and their dominant influencing factors employing linear mixed-effects models, variance partitioning method, piecewise Structural Equation Modeling, etc. The results show that with increasing elevation, SLA significantly decreases, and LDMC significantly increases (P < 0.001). This indicates different resource utilization strategies of grassland plants across elevation gradients, transitioning from a "faster investment-return" at lower elevations to a "slower investment-return" at higher elevations. Across different elevation gradients, climatic factors are the main factors affecting grassland plant resource utilization strategies, with soil nutrient factors also playing a non-negligible coordinating role. Among these, mean annual precipitation and hottest month mean temperature are key climatic factors influencing SLA of grassland plants, explaining 28.94% and 23.88% of SLA variation, respectively. The key factors affecting LDMC of grassland plants are mainly hottest month mean temperature and soil phosphorus content, with relative importance of 24.24% and 20.27%, respectively. Additionally, the direct effect of elevation on grassland plant resource utilization strategies is greater than its indirect effect (through influencing climatic and soil nutrient factors). These findings emphasize the substantive impact of elevation on grassland plant resource utilization strategies and have important ecological value for grassland management and protection under global change.

The "Leafing Intensity Premium" Hypothesis and the Scaling Relationships of the Functional Traits of Bamboo Species.

The "leafing intensity premium" hypothesis proposes that leaf size results from natural selection acting on different leafing intensities, i.e., the number of leaves per unit shoot volume or mass. The scaling relationships among various above-ground functional traits in the context of this hypothesis are important for understanding plant growth and ecology. Yet, they have not been sufficiently studied. In this study, we selected four bamboo species of the genus Indocalamus Nakai and measured the total leaf fresh mass per culm, total non-leaf above-ground fresh mass, total number of leaves per culm, and above-ground culm height of 90 culms from each species. These data were used to calculate leafing intensity (i.e., the total number of leaves per culm divided by the total non-leaf above-ground fresh mass) and mean leaf fresh mass per culm (i.e., the total leaf fresh mass per culm divided by the total number of leaves per culm). Reduced major axis regression protocols were then used to determine the scaling relationships among the various above-ground functional traits and leafing intensity. Among the four species, three exhibited an isometric (one-to-one) relationship between the total leaf fresh mass per culm and the total non-leaf above-ground fresh mass, whereas one species (Indocalamus pumilus) exhibited an allometric (not one-to-one) relationship. A negative isometric relationship was found between the mean leaf fresh mass per culm and the leafing intensity for one species (Indocalamus pedalis), whereas three negative allometric relationships between mean leaf fresh mass per culm and leafing intensity were observed for the other three species and the pooled data. An exploration of the alternative definitions of "leafing intensity" showed that the total number of leaves per culm divided by the above-ground culm height is superior because it facilitates the non-destructive calculation of leafing intensity for Indocalamus species. These results not only confirm the leafing intensity premium hypothesis for bamboo species but also highlight the interconnected scaling relationships among different functional traits, thereby contributing to our understanding of the ecological and evolutionary significance of leaf size variation and biomass investment strategies.