Temporal variations in absorption and translocation of heavy metal(loid)s in pak choi (Brassica rapa L.) under open-field and greenhouse cultivation.

Elucidating the absorption and translocation of heavy metal(loid)s by common vegetables across different growth environments and stages is crucial for conducting accurate environmental risk assessments and for associated control. This study investigated temporal variations in the absorption and translocation capacities of pak choi (Brassica rapa L.) for As, Cd, Cr, Cu, Pb, and Zn in polluted soils during the plant growth cycle under greenhouse and open-field cultivation modes. Results showed high root metal(loid) bioconcentration factors and root-to-shoot translocation factors for Cd (0.25 and 1.44, respectively) and Zn (0.26 and 1.01), but low values for As (0.06 and 0.88) and Pb (0.06 and 0.87). The Cd concentration in the aerial edible parts peaked during the early slow growth period, whereas other heavy metal(loid)s peaked during the later stable maturity period. Root bioconcentration and root-to-shoot translocation factors did not significantly differ between cultivation modes. However, greenhouse cultivation exhibited lower average Cd and Zn concentrations in the edible parts and cumulative uptake amounts of most metal(loid)s than open-field cultivation during the typical harvest period spanning days 60 and 90. Short-term transitioning from open-field to greenhouse cultivation may reduce health risks associated with heavy metal(loid) intake via pak choi consumption. These findings facilitate sustainable agricultural practices and food safety management.

Moderate oxidative stress promotes epithelial-mesenchymal transition in the lens epithelial cells via the TGF-ß/Smad and Wnt/ß-catenin pathways.

The epithelial-mesenchymal transition (EMT) plays a significant role in fibrosis and migration of lens epithelial cells (LECs), and eventually induces posterior capsule opacification (PCO). In the past, it was generally believed that the TGF-ß/Smad pathway regulates lens EMT. A recent study found that attenuated glutathione level promotes LECs EMT via the Wnt/ß-catenin pathway, which suggests a more complex pathogenesis of PCO. To test the hypothesis, we used the mouse cataract surgery PCO model and tested both canonical Wnt/ß-catenin and TGF-ß/Smad signaling pathways. The results showed that both TGF-ß/Smad and Wnt/ß-catenin pathways were activated during the lens capsule fibrosis. Compared with the freshly isolated posterior capsule, the expression level of phosphorylated Smad2 was highest at day3 and then slightly decreased, but the expression level of Wnt10a gradually increased from day0 to day7. It shows that these two pathways are involved in the lens epithelium's fibrotic process and may play different roles in different periods. Subsequently, we established oxidative stress-induced EMT model in primary porcine lens epithelial cells and found that both the TGF-ß/Smad and Wnt/ß-catenin pathways were activated. Further study suggests that block Wnt/ß-catenin pathway using XAV939 alone or block TGF-ß/Smad pathway using LY2109761 could partially block pLECs fibrosis, but blocking Wnt/ß-catenin and TGF-ß/Smad pathway using combined XAV939 and LY2109761 could completely block pLECs fibrosis. In conclusion, this study demonstrates that both TGF-ß/Smad and canonical Wnt/ß-catenin pathways play a significant role in regulating epithelial-mesenchymal transformation of lens epithelial cells but might be in a different stage.

Activation of liver X receptor α protects amyloid ß1-40 induced inflammatory and senescent responses in human retinal pigment epithelial cells.

OBJECTIVE: To investigate whether activation of the liver X receptors (LXRs) inhibits amyloid ß1-40 (Aß1-40) induced inflammatory and senescent responses in human retinal pigment epithelial (RPE) cells. MATERIALS AND METHODS: Confluent cultures of human primary RPE and ARPE-19 cells pretreated with 5 µΜ of TO901317 (TO90), a synthetic agonist of LXR, or vehicle were incubated with 1 µΜ of Aß1-40 or Aß40-1. The optimum concentrations of Aß1-40 and TO90 were determined by cell viability assay. Pro-inflammatory cytokines IL-6, IL-8, MCP-1 were detected by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Expression and localization of an aging protein p16INK4a (p16) were analyzed by western blotting and immunofluorescence. Expressions of LXRs and one of their target genes ATP-binding cassette transporter A1 (ABCA1) were examined by real-time PCR and western blotting. Phosphorylated transcription inhibition factor-κB-α (p-IκB-α) was assessed by western blotting. RESULTS: A negative linear relationship between the Aß1-40 concentration and the cell viability was evident, indicating Aß1-40 decreased ARPE-19 cell viability in a dose-dependent manner. Aß1-40 enhanced the expression of IL-6, IL-8, MCP-1 as well as p16 in both RPE cell lines at both mRNA and protein levels, whereas TO90 counteracted the detrimental effects. TO90 upregulated the expression of LXRα and its target gene ABCA1, but it did not affect the expression of LXRß. Meanwhile, TO90 inhibited the phosphorylation of IκB-α mediated by Aß1-40 stimulation. CONCLUSION: Activation of the LXRα-ABCA1 axis may alleviate Aß1-40 induced inflammatory and senescent responses in RPE cells. The beneficial effect appears associated with the inhibition of the NF-κB signaling pathway.

[Spatiotemporal Response of Ecosystem Service Value to Land Use Change in the Lanzhou-Xining Urban Agglomeration over the Past 20 Years].

With rapid urbanization and human activities exacerbating threats to the degradation of various ecosystem services in modern urban agglomerations, the exploration of the state of ecological security at the scale of urban agglomerations is of great significance. This study considered the Lanzhou-Xining Urban Agglomeration as the research area, based on the land use data in 2000, 2005, 2010, 2015, and 2020. At the same time, the landscape ecological risk index was introduced. The land use change characteristics of the Lanzhou-Xining Urban Agglomeration were analyzed by using the land use transfer matrix, the value per unit area equivalent factor method, and the bivariate spatial autocorrelation analysis method to elucidate the impacts of the changes in the ecological risk index induced by the land use transition on the value of ecosystem services. This study analyzed the land use change characteristics of the Lanzhou-Xining Urban Agglomeration and elucidated the impacts of changes in the ecological risk index on the value of ecosystem services caused by land use transformation. The results showed that:① During the period from 2000 to 2020, the land use types of the Lanzhou-Xining Urban Agglomeration were mainly dominated by grassland, cropland, and forest land. The construction land area had expanded significantly mainly from cropland and grassland, and the six land use types had strong cross-transformation. The total area of land use change was 6 646.05 km2. ② In terms of spatial changes, the spatial pattern of ecosystem service value in the Lanzhou-Xining Urban Agglomeration had not undergone obvious transformation. However, the regional variability was significant, generally showing the distribution characteristics of high in the northwest and low in the southeast. ③From the perspective of temporal change, the value of ecosystem services in the Lanzhou-Xining Urban Agglomeration showed an upward trend, with the total flow of value increasing from 186.459 billion yuan to 192.156 billion yuan, with a total value-added of 5.697 billion yuan. ④ There was a rising trend in the overall ecological risk index of the Lanzhou-Xining Urban Agglomeration over the past 20 years. Low ecological risk areas and lower ecological risk areas dominated the ecological risk areas. There was a significant positive correlation between the value of ecosystem services and the ecological risk index. This study aimed to reveal the understanding of the impacts of land-use practices on ecosystem service values and ecological risks, to provide important references for regional ecological risk management and land-use policy formulation, and thus to promote the high-quality development of the ecological environment in the Yellow River Basin.

Seed rain composition responds to climate change in a subtropical forest.

Recent climate change has been shown to alter aspects of forest plant demography, such as growth and mortality, but less attention has been focused on how climate change alters the reproduction of plant populations through time. We hypothesized that the plant seed production would respond to climate change, and that the response would differ according to plant life form and functional traits. We tested this hypothesis by examining climate change from 2005 to 2020 and by determining the temporal trends of seed rain and seed production from plants with different life forms (e.g., herbs, vines, trees, palms) and of tree species with different statures as well as leaf, seed and wood traits during 2014-2020. We also tested the correlation between meteorological variables and time series of seed rain using cross correlation analysis. We found increasing wetness (lower vapor pressure deficit) through time but with decreasing minimum relative humidity, which is a pattern consistent with trends seen in many other parts of the world. During the study period, seed production of shrubs and relative contribution of woody vines to total seed rain decreased, while relative contribution of palms to total seed rain and tree species with more conservative leaf traits increased their contribution to total seed rain. Overall, these trends were well explained by the trends of meteorological variables and the responses of these life forms to climate change in previous studies. Additionally, the increasingly conservative leaf traits were also consistent with shifts in traits following recovery from disturbance. Our results suggest that a trait-based approach may help to unveil trends that are not readily apparent by examining seed counts alone. The compositional change found in the seed rain may indicate future shifts in forest species composition and should be incorporated into future studies of forest modelling and projections under climate change.

The complete chloroplast genome and phylogenomic analysis of Camellia sinensis var. sinensis cultivar 'Liupao', a landrace from Guangxi, China.

Liupao tea is one of the well-known Chinese tea brands and a famous local specialty in Wuzhou, Guangxi, China. However, the genetic background and phylogenetic relationship of the native resource plants of Liupao tea need study, especially at the genomic level. In this study, we reported the complete chloroplast (cp) genome sequence of Camellia sinensis var. sinensis cultivar 'Liupao' (LP, Liupao tea population) and inferred its phylogenetic relationship to other tea plant variants or cultivars. The cp genome had a total length of 157,097 bp and the overall GC content was 37.3%. The cp genome contained one LSC region (86,641 bp) and one SSC region (18,276 bp), which were separated by two IR regions (26,090 bp, respectively). Moreover, the cp genomes were composed of 130 genes, including 86 protein-coding genes, 36 tRNA genes, and eight rRNA genes. The phylogenetic analysis showed that LP was closely related to C. sinensis var. pabilimba cv. 'Lingyunbaihao'. This study will provide useful information for further investigating the genetic background, evolution, and breeding of LP as well as other tea cultivars and varieties.

[Temporal and Spatial Distributions of O3 Concentration and Potential Source Area Analysis of Hexi Corridor Based on Satellite and Ground Monitoring].

Based on 2005-2020 O3 column concentration data of OMI remote sensing satellite, combined with air pollutant data from 10 nationally controlled environmental automatic monitoring stations in the Hexi Corridor and global data assimilation system meteorological data, we used Kriging interpolation, correlation analysis, and backward trajectory (HYSPLIT) models to explore the temporal and spatial distribution characteristics, meteorological factors, transmission paths, and potential sources of O3 in the Hexi Corridor. The results showed the following:① in terms of temporal distribution, O3 column concentration showed an upward trend in 2005-2010 and 2014-2020 and downward trend in 2010-2014; the maximum and minimum values were reached in 2010 and 2014 (332.31 DU and 301.00 DU), respectively, and seasonal changes showed that those in spring and winter were significantly higher than those in summer and autumn. ② In terms of spatial distribution, O3 column concentration showed a latitudinal band distribution characteristic of increasing from southwest to northeast; the high-value areas were primarily distributed in urban areas with low terrain, and the median zone was latitudinally striped with the basic alignment of the Qilian foothills. ③ The analysis of meteorological conditions revealed that temperature, wind speed, and sunshine hours were positively correlated with O3, and relative humidity was negatively correlated with O3. ④ By simulating the airflow transportation trajectory of the receiving point in Wuwei City, it was found that the direction of the O3 conveying path was relatively singular; the dominant airflow in each season was primarily in the west and northwest; and the proportions were 71.62%, 66.85%, 61.22%, and 77.78%, respectively. There were certain seasonal differences in the source areas of O3 potential contribution:the high-value areas of O3 potential sources in spring, summer, and autumn were distributed in Baiyin City and Lanzhou City, which were southeast wind sources, and the high-value areas in winter were distributed between the Badain Jaran Desert and the Tengger Desert, which was the north wind source.

Semi-interpenetrating anion exchange membranes using hydrophobic microporous linear poly(ether ketone).

In order to realise high ionic conductivity and improved chemical stability, a series of anion exchange membranes (AEMs) with semi-interpenetrating polymer network (sIPN) has been prepared via the incorporation of crosslinked poly(biphenyl N-methylpiperidine) (PBP) and spirobisindane-based intrinsically microporous poly(ether ketone) (PEK-SBI). The formation of phase separated structures as a result of the incompatibility between the hydrophilic PBP network and the hydrophobic PEK-SBI segment, has successfully promoted the hydroxide ion conductivity of AEMs. A swelling ratio (SR) as low as 12.2 % at 80 °C was recorded for the sIPN containing hydrophobic PEK-SBI as the linear polymer and crosslinked structure with a mass ratio of PBP to PEK-SBI of 90/10 (sIPN-90/10(PEK-SBI)). The sIPN-90/10(PEK-SBI) AEM achieved the highest hydroxide ion conductivity of 122.4 mS cm-1 at 80 °C and a recorded ion exchange capacity (IEC) of 2.26 meq g-1. Atomic force microscopy (AFM) and transmission electron microscopy (TEM) clearly revealed the improved phase separation structure of sIPN-90/10(PEK-SBI). N2 adsorption isotherm indicated that the Brunauer-Emmett-Teller (BET) surface area of the AEMs increased with the increase of microporous PEK-SBI content. Interestingly, the sIPN-90/10(PEK-SBI) AEM showed good alkaline stability for being able to maintain a conductivity of 94.7 % despite being soaked in a 1 M sodium hydroxide solution at 80 °C for 30 days. Meanwhile, a peak power density of 481 mW cm-2 can be achieved by the hydrogen/oxygen single cell using sIPN-90/10(PEK-SBI) as the AEM.

Neutrophil extracellular traps and complications of liver transplantation.

Many end-stage liver disease etiologies are attributed to robust inflammatory cell recruitment. Neutrophils play an important role in inflammatory infiltration and neutrophil phagocytosis, oxidative burst, and degranulation. It has also been suggested that neutrophils may release neutrophil extracellular traps (NETs) to kill pathogens. It has been proven that neutrophil infiltration within the liver contributes to an inflammatory microenvironment and immune cell activation. Growing evidence implies that NETs are involved in the progression of numerous complications of liver transplantation, including ischemia-reperfusion injury, acute rejection, thrombosis, and hepatocellular carcinoma recurrence. NETs are discussed in this comprehensive review, focusing on their effects on liver transplantation complications. Furthermore, we discuss NETs as potential targets for liver transplantation therapy.

Case report: A case of severe acute hepatitis of unknown origin.

According to analyses of etiology, clinical features, diagnostic methods, and treatment strategies by summarizing a case of unexplained acute hepatitis recently experienced, we are aiming to provide some information to enrich the clinical experience in diagnosis and treatment of severe acute hepatitis of unknown etiology in young children. A boy, aged 10 years and 6 months old, was admitted to the hospital due to acute abdominal pain, jaundice, and exceptionally high levels of ALT and AST. A range of measures, including patient history, physical examination, and routine laboratory testing, were performed. Furthermore, strategies such as trio-based next-generation sequencing (Trio-NGS) and liver biopsy, as well as metagenomic NGS (mNGS) of blood and liver samples were also performed. In summary, this case was an acute severe non-A-E hepatitis that is a probable case with hepatitis of unknown origin. Immunohistochemical analysis showed an immune injury in liver tissues. Torque teno virus (TTV) sequences were detected by mNGS assay. As for treatment strategies, in addition to general treatment, this patient also underwent plasmapheresis and methylprednisolone treatment due to disease deterioration. The patient's liver function was improved afterward and discharged after one month of treatment. Taken together, this work reported the clinical feature and treatment of severe acute hepatitis with non-A-E hepatitis in detail. The potential mechanism of liver damage might be due to an immune attack in which TTV might play a role as a co-factor.

Elevated YKL-40 serum levels may contribute to wet age-related macular degeneration via the ERK1/2 pathway.

Choroidal neovascularization (CNV) is a key characteristic of wet age-related macular degeneration (AMD) that can lead to severe vision loss in the elderly. Anti-VEGF therapy is currently the premier strategy for wet AMD, but it has limited efficacy. Previous studies have shown that chitinase-3-like-1 (YKL-40) can promote microangiogenesis and inflammation, but its effect on CNV formation has not yet been studied. Here, we investigated the potential role of YKL-40 in wet AMD and the underlying mechanism(s). We report that the serum expression of YKL-40 in wet AMD patients was significantly higher than that in control patients and was positively correlated with VEGF expression, indicating that YKL-40 may participate in the development of wet AMD. In addition, YKL-40 and VEGF expression levels were observed to be increased and the ERK1/2 pathway activated in the neuroretinal (NR) and RPE/choroid tissues of mice with laser-induced CNV. The YKL-40 and phosphorylated protein levels of the ERK1/2 pathway were decreased after intravitreal injection with an anti-YKL-40 antibody, suggesting that anti-YKL-40 could inhibit the activation of the ERK1/2 pathway. These results indicate that YKL-40 may serve as a novel target for the diagnosis and treatment of wet AMD.

Interaction of imidacloprid with hemoglobin by fluorescence and circular dichroism.

Imidacloprid belongs to a major new class of insecticides, called neonicotinoids, which are accounting for 11-15% of the total insecticide market. The binding characteristics of insecticide imidacloprid with hemoglobin (Hb) have been studied by employing different spectroscopic techniques. The results proved the formation of complex between imidacloprid and Hb. Hydrophobic interaction and hydrogen bond dominated in the association reaction. Hydrophobic probe 8-anilino-1-naphthalenesulfonic acid (ANS) competitive experiments indicated that the binding of imidacloprid to Hb primarily took place in hydrophobic regions. The distance between Hb donor and acceptor imidacloprid was 4.88 nm as derived from Förster's theory. Alternations of Hb secondary structure in the presence of imidacloprid were confirmed by synchronous fluorescence, circular dichroism (CD) and three-dimensional fluorescence spectra. This study enriches our understanding of toxic effect of imidacloprid to the physiologically important protein Hb.

Increased serum levels of soluble CD146 and vascular endothelial growth factor receptor 2 in patients with exudative age-related macular degeneration.

AIM: To investigate serum levels of soluble CD146 (sCD146) and vascular endothelial growth factor receptor 2 (VEGFR2) in patients with age-related macular degeneration (AMD). METHODS: Eighty-eight patients with exudative AMD and 45 sex- and age-matched healthy controls were enrolled in this study conducted in China. Serum samples was obtained from the patients with exudative AMD and from the controls. Serum sCD146 and VEGFR2 protein levels were measured using an enzyme-linked immunosorbent assay. RESULTS: We found that serum sCD146 and VEGFR2 protein levels were significantly higher in the patients with exudative AMD group than in the controls (t=3.859, P<0.001 and t=3.829, P<0.001, respectively). Serum sCD146 levels were significantly higher in patients with classic choroidal neovascularization (CNV) than in those with occult CNV (t=9.899, P<0.001). There was a significant difference in the trend for exudative AMD in the highest versus lowest quartile of circulating sCD146 levels (χ 2=10.29, P=0.001). The receiver operating characteristic curve analysis showed that the area under the curve was 0.696 for sCD146 (95%CI: 0.601-0.791) with an optimum diagnostic cut-off value of 157.16 ng/mL, a sensitivity of 55.7%, and a specificity of 82.2%. CONCLUSION: The serum sCD146 level increases and may be a biomarker for exudative AMD.

Testing the competition-colonization trade-off and its correlations with functional trait variations among subtropical tree species.

The competition-colonization trade-off, by which species can partition spatial niches, is a potentially important mechanism allowing the maintenance of species diversity in plant communities. We examined whether there was evidence for this trade-off among tree species in a subtropical forest and how it correlated with eight functional traits. We developed and estimated a metric for colonization ability that incorporates both fecundity and seed dispersal based on seed trap data and the sizes and distributions of adult trees. Competitive ability was estimated as survival probability under high crowding conditions based on neighborhood models. Although we found no significant relationship between colonization and competitive abilities, there was a significant negative correlation between long distance dispersal ability and competitive ability at the 5 cm size class. Colonizers had traits associated with faster growth, such as large leaves and low leaf lamina density, whereas competitors had traits associated with higher survival, such as dense wood. Our results imply that any trade-off between competition and colonization may be more determined by dispersal ability than by fecundity, suggesting that seed dispersal is an important contributor to diversity maintenance. Future work should test how competitive ability covaries with the components of colonization ability, as we did here.

Elastic-Plastic Transformation of Polyelectrolyte Complex Hydrogels from Chitosan and Sodium Hyaluronate.

Hydrogels formed by polyelectrolyte complexation (PEC) of oppositely charged biopolymers, free of any chemical additives, are promising biomaterials. In this work, the mechanical behavior of hydrogels consisting of positively charged chitosan and negatively charged sodium hyaluronate (HA) at balanced charge composition is investigated. These hydrogels exhibit strong tensile strain and strain rate dependence. They are elastic-like, independent of the strain rate at small strain, but exhibit plastic-like behavior above the yield point by showing a monotonous decrease of the stress. The cyclic tensile test demonstrates that these hydrogels exhibit small and quickly recoverable hysteresis in the elastic-like region, but large and partially recoverable hysteresis above the yield point. The stress relaxation experiment shows a plateau in the reduced stress followed by an abrupt fracture, and the time-to-failure decreases exponentially with increasing applied step-strain. Such elastic-to-plastic-like transformation of the biopolymer PEC gels is quite different from the behaviors of PEC hydrogels formed by oppositely charged vinyl-type synthetic polyelectrolytes due to the difference in flexibility, charge density, and ionic bond strength of these polymers.

Different relationships between temporal phylogenetic turnover and phylogenetic similarity and in two forests were detected by a new null model.

BACKGROUND: Ecologists have been monitoring community dynamics with the purpose of understanding the rates and causes of community change. However, there is a lack of monitoring of community dynamics from the perspective of phylogeny. METHODS/PRINCIPLE FINDINGS: We attempted to understand temporal phylogenetic turnover in a 50 ha tropical forest (Barro Colorado Island, BCI) and a 20 ha subtropical forest (Dinghushan in southern China, DHS). To obtain temporal phylogenetic turnover under random conditions, two null models were used. The first shuffled names of species that are widely used in community phylogenetic analyses. The second simulated demographic processes with careful consideration on the variation in dispersal ability among species and the variations in mortality both among species and among size classes. With the two models, we tested the relationships between temporal phylogenetic turnover and phylogenetic similarity at different spatial scales in the two forests. Results were more consistent with previous findings using the second null model suggesting that the second null model is more appropriate for our purposes. With the second null model, a significantly positive relationship was detected between phylogenetic turnover and phylogenetic similarity in BCI at a 10 m×10 m scale, potentially indicating phylogenetic density dependence. This relationship in DHS was significantly negative at three of five spatial scales. This could indicate abiotic filtering processes for community assembly. Using variation partitioning, we found phylogenetic similarity contributed to variation in temporal phylogenetic turnover in the DHS plot but not in BCI plot. CONCLUSIONS/SIGNIFICANCE: The mechanisms for community assembly in BCI and DHS vary from phylogenetic perspective. Only the second null model detected this difference indicating the importance of choosing a proper null model.

Unimodal tree size distributions possibly result from relatively strong conservatism in intermediate size classes.

Tree size distributions have long been of interest to ecologists and foresters because they reflect fundamental demographic processes. Previous studies have assumed that size distributions are often associated with population trends or with the degree of shade tolerance. We tested these associations for 31 tree species in a 20 ha plot in a Dinghushan south subtropical forest in China. These species varied widely in growth form and shade-tolerance. We used 2005 and 2010 census data from that plot. We found that 23 species had reversed J shaped size distributions, and eight species had unimodal size distributions in 2005. On average, modal species had lower recruitment rates than reversed J species, while showing no significant difference in mortality rates, per capita population growth rates or shade-tolerance. We compared the observed size distributions with the equilibrium distributions projected from observed size-dependent growth and mortality. We found that observed distributions generally had the same shape as predicted equilibrium distributions in both unimodal and reversed J species, but there were statistically significant, important quantitative differences between observed and projected equilibrium size distributions in most species, suggesting that these populations are not at equilibrium and that this forest is changing over time. Almost all modal species had U-shaped size-dependent mortality and/or growth functions, with turning points of both mortality and growth at intermediate size classes close to the peak in the size distribution. These results show that modal size distributions do not necessarily indicate either population decline or shade-intolerance. Instead, the modal species in our study were characterized by a life history strategy of relatively strong conservatism in an intermediate size class, leading to very low growth and mortality in that size class, and thus to a peak in the size distribution at intermediate sizes.

Endoscopic screening and determination of p53 and proliferating cell nuclear antigen in esophageal multistage carcinogenesis: a comparative study between high- and low-risk populations in Henan, northern China.

The objective of this study was to characterize the histologic changes from endoscopic screening for early esophageal cancer (EC) on subjects at high-incidence area (HIA) and low-incidence area (LIA) in Henan, China, and to further compare the changes in p53 and proliferating cell nuclear antigen (PCNA) in the multistage of human esophageal carcinogenesis from these two populations. The detection rate of basal cell hyperplasia (BCH) and dysplasia (DYS) was higher in the subjects from HIA than in those from LIA. Out of the 1568 symptom-free subjects examined at HIA, 10 (0.6%) cases with early squamous cell carcinoma (SCC) were identified. Immunoreactivity of p53 and PCNA was observed in cell nuclei of esophageal biopsies and surgically resected esophageal cancer specimens both in HIA and LIA. With the lesions progressed from normal epithelium to BCH to DYS to SCC, the positive-immunostaining cells expanded from basal layer to superficial layer, and the number of positive cells/mm2 for p53 and PCNA increased, and was significantly higher in HIA than in LIA among the similar morphological lesions (P < 0.01). The number of p53 positive cells/mm2 in SCC from HIA was almost fivefold higher than SCC from LIA (P < 0.01). The remarkable difference was also observed between HIA and LIA in DYS and BCH. The present results indicate that p53 protein accumulation is an important early biomarker for identifying high-risk subjects for EC.