Lyophilized lymph nodes for improved delivery of chimeric antigen receptor T cells.

Lymph nodes are crucial organs of the adaptive immune system, orchestrating T cell priming, activation and tolerance. T cell activity and function are highly regulated by lymph nodes, which have a unique structure harbouring distinct cells that work together to detect and respond to pathogen-derived antigens. Here we show that implanted patient-derived freeze-dried lymph nodes loaded with chimeric antigen receptor T cells improve delivery to solid tumours and inhibit tumour recurrence after surgery. Chimeric antigen receptor T cells can be effectively loaded into lyophilized lymph nodes, whose unaltered meshwork and cytokine and chemokine contents promote chimeric antigen receptor T cell viability and activation. In mouse models of cell-line-derived human cervical cancer and patient-derived pancreatic cancer, delivery of chimeric antigen receptor T cells targeting mesothelin via the freeze-dried lymph nodes is more effective in preventing tumour recurrence when compared to hydrogels containing T-cell-supporting cytokines. This tissue-mediated cell delivery strategy holds promise for controlled release of various cells and therapeutics with long-term activity and augmented function.

Lymph node-biomimetic scaffold boosts CAR-T therapy against solid tumor.

The limited infiltration and persistence of chimeric antigen receptor (CAR)-T cells is primarily responsible for their treatment deficits in solid tumors. Here, we present a three-dimensional scaffold, inspired by the physiological process of T-cell proliferation in lymph nodes. This scaffold gathers the function of loading, delivery, activation and expansion for CAR-T cells to enhance their therapeutic effects on solid tumors. This porous device is made from poly(lactic-co-glycolic acid) by a microfluidic technique with the modification of T-cell stimulatory signals, including anti-CD3, anti-CD28 antibodies, as well as cytokines. This scaffold fosters a 50-fold CAR-T cell expansion in vitro and a 15-fold cell expansion in vivo. Particularly, it maintains long-lasting expansion of CAR-T cells for up to 30 days in a cervical tumor model and significantly inhibits the tumor growth. This biomimetic delivery strategy provides a versatile platform of cell delivery and activation for CAR-T cells in treating solid tumors.

Cryo-shocked tumor cells deliver CRISPR-Cas9 for lung cancer regression by synthetic lethality.

Although CRISPR-mediated genome editing holds promise for cancer therapy, inadequate tumor targeting and potential off-target side effects hamper its outcomes. In this study, we present a strategy using cryo-shocked lung tumor cells as a CRISPR-Cas9 delivery system for cyclin-dependent kinase 4 (CDK4) gene editing, which initiates synthetic lethal in KRAS-mutant non-small cell lung cancer (NSCLC). By rapidly liquid nitrogen shocking, we effectively eliminate the pathogenicity of tumor cells while preserving their structure and surface receptor activity. This delivery system enables the loaded CRISPR-Cas9 to efficiently target to lung through the capture in pulmonary capillaries and interactions with endothelial cells. In a NSCLC-bearing mouse model, the drug accumulation is increased nearly fourfold in lung, and intratumoral CDK4 expression is substantially down-regulated compared to CRISPR-Cas9 lipofectamine nanoparticles administration. Furthermore, CRISPR-Cas9 editing-mediated CDK4 ablation triggers synthetic lethal in KRAS-mutant NSCLC and prolongs the survival of mice.

Geographical patterns and determinants of insect biodiversity in China.

Insects play important roles in the maintenance of ecosystem functioning and the provision of livelihoods for millions of people. However, compared with terrestrial vertebrates and angiosperms, such as the giant panda, crested ibis, and the metasequoia, insect conservation has not attracted enough attention, and a basic understanding of the geographical biodiversity patterns for major components of insects in China is lacking. Herein, we investigated the geographical distribution of insect biodiversity across multiple dimensions (taxonomic, genetic, and phylogenetic diversity) based on the spatial distribution and molecular DNA sequencing data of insects. Our analysis included 18 orders, 360 families, 5,275 genera, and 14,115 species of insects. The results revealed that Southwestern and Southeastern China harbored higher insect biodiversity and numerous older lineages, representing a museum, whereas regions located in Northwestern China harbored lower insect biodiversity and younger lineages, serving as an evolutionary cradle. We also observed that mean annual temperature and precipitation had significantly positive effects, whereas altitude had significantly negative effects on insect biodiversity in most cases. Moreover, cultivated vegetation harbored the highest insect taxonomic and phylogenetic diversity, and needleleaf and broadleaf mixed forests harbored the highest insect genetic diversity. These results indicated that human activities may positively contribute to insect spatial diversity on a regional scale. Our study fills a knowledge gap in insect spatial diversity in China. These findings could help guide national-level conservation plans and the post-2020 biodiversity conservation framework.

Effects of hydroxy methionine zinc on growth performance, immune response, antioxidant capacity, and intestinal microbiota of red claw crayfish (Procambarus clarkii).

This study aimed to evaluate the effects of varying zinc (Zn) levels on the growth performance, non-specific immune response, antioxidant capacity, and intestinal microbiota of red claw crayfish (Procambarus clarkii (P. clarkii)). Adopting hydroxy methionine zinc (Zn-MHA) as the Zn source, 180 healthy crayfish with an initial body mass of 6.50 ± 0.05 g were randomly divided into the following five groups: X1 (control group) and groups X2, X3, X4, and X5, which were fed the basal feed supplemented with Zn-MHA with 0, 15, 30, 60, and 90 mg kg-1, respectively. The results indicated that following the addition of various concentrations of Zn-MHA to the diet, the following was observed: Specific growth rate (SGR), weight gain rate (WGR), total protein (TP), total cholesterol (TC), the activities of alkaline phosphatase (AKP), phenoloxidase (PO), total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD) and catalase (CAT), the expression of CTL, GPX, and CuZn-SOD genes demonstrated a trend of rising and then declining-with a maximum value in group X4-which was significantly higher than that in group X1 (P < 0.05). Zn deposition in the intestine and hepatopancreas, the activity of GSH-PX, and the expression of GSH-PX were increased, exhibiting the highest value in group X5. The malonaldehyde (MDA) content was significantly reduced, with the lowest value in group X4, and the MDA content of the Zn-MHA addition groups were significantly lower than the control group (P < 0.05). In the analysis of the intestinal microbiota of P. clarkii, the number of operational taxonomic units in group X4 was the highest, and the richness and diversity indexes of groups X3 and X4 were significantly higher than those in group X1 (P < 0.05). Meanwhile, the dietary addition of Zn-MHA decreased and increased the relative abundance of Proteobacteria and Tenericutes, respectively. These findings indicate that supplementation of dietary Zn-MHA at an optimum dose of 60 mg kg-1 may effectively improve growth performance, immune response, antioxidant capacity, and intestinal microbiota richness and species diversity in crayfish.

Influence of Pasteurization on Maillard Reaction in Lactose-Free Milk.

In order to improve the safety and quality of lactose-free milk (LFM) Maillard reaction products (MRPs), this study used raw cow's milk as raw material and lactase hydrolysis to prepare LFM, which was heat-treated using pasteurization and then placed in storage temperatures of 4 °C, 25 °C and 37 °C to investigate the changes in the Maillard reaction (MR). The results of the orthogonal test showed that the optimal conditions for the hydrolysis of LFM are as follows: the hydrolysis temperature was 38 °C, the addition of lactase was 0.03%, and the hydrolysis time was 2.5 h. Under these conditions, the lactose hydrolysis rate reached 97.08%, and the lactose residue was only 0.15 g/100 g as determined by high-performance liquid chromatography (HPLC), complying with the standard of LFM in GB 28050-2011. The contents of furoamic acid and 5-hydroxymethylfurfural were determined by high-performance liquid chromatography, the color difference was determined by CR-400 color difference meter, and the internal fluorescence spectrum was determined by F-320 fluorescence spectrophotometer. The test results showed that the variation range of furosine in lactose-free milk after pasteurization was 44.56~136.45 mg/100g protein, the range of 5-hydroxymethylfurfural (HMF) was 12.51~16.83 mg/kg, the color difference ranges from 88.11 to 102.53 in L*, from -0.83 to -0.10 in a*, and from 1.88 to 5.47 in b*. The furosine content of LFM during storage at 4, 25, and 37 °C ranged from 44.56 to 167.85, 44.56 to 287.13, and 44.56 to 283.72 mg/100 g protein, respectively. The average daily increase in protein content was 1.18-3.93, 6.46-18.73, and 15.7-37.66 mg/100 g, respectively. The variation range of HMF was 12.51~17.61, 12.51~23.38, and 12.51~21.1 mg/kg, and the average daily increase content was 0.03~0.07, 0.47~0.68, and 0.51~0.97 mg/kg, respectively. During storage at 4 °C, the color difference of LFM ranged from 86.82 to 103.82, a* ranged from -1.17 to -0.04, and b* ranged from 1.47 to 5.70. At 25 °C, color difference L* ranges from 72.09 to 102.35, a* ranges from -1.60 to -0.03, b* ranges from 1.27 to 6.13, and at 37 °C, color difference L* ranges from 58.84 to 102.35, a* ranges from -2.65 to 1.66, and b* ranges from 0.54 to 5.99. The maximum fluorescence intensity (FI) of LFM varies from 131.13 to 173.97, 59.46 to 173.97, and 29.83 to 173.97 at 4, 25, and 37 °C. In order to reduce the effect of the Maillard reaction on LFM, it is recommended to pasteurize it at 70 °C-15 s and drink it as soon as possible during the shelf life within 4 °C.

Habitat-specific conservation priorities of multidimensional diversity patterns of amphibians in China effectively contribute to the '3030' target.

Amphibia is the most threatened animal group among all land vertebrates in the context of anthropogenic global change. Filling the conservation gaps for this taxonomic group could help achieve the ambitious target of covering 30 % of the land by 2030 ('3030' target) set by the 15-th meeting of the Conference of the Parties (COP15). In this study, we compiled the most up-to-date occurrence records and corresponding species-specific traits and phylogenies of amphibians in China (particularly those newly described in the past decade) to explore the spatial distribution patterns of multidimensional diversity (including taxonomic, functional, and phylogenetic) for different species groups (including all, endemic and threatened). Additionally, a new conservation gap index (CGI) was proposed and applied to the analysis of multi-objective conservation strategies. The results showed that the spatial distribution of taxonomic, functional and phylogenetic diversity of amphibians in China is markedly geographically diverse, with common hotspots for all three concentrated in the humid mountainous regions of southern China. The CGI, which is independent of arbitrary threshold selection and grid cell size, showed that the conservation gap for amphibians in China is largest in biomes such as tropical and subtropical moist broadleaf forests and temperate broadleaf and mixed forests. The multi-objective conservation analysis revealed that the Yangtze River basin, Pearl River basin and Southeast Basin in China have pivotal roles in achieving the '3030' target due to their high taxonomic, phylogenetic and functional diversity, relatively high proportion of threatened and endemic species, and low coverage of existing nature reserves. Notably, sustainable management of less-protected habitats, including farmlands and grasslands, can reduce the area requirement of strict protection for reaching the '3030' conservation goal. This study provides practical strategies for guiding amphibian conservation by systematically integrating multidimensional biodiversity information, habitat features and the spatial distributions of the existing natural reserves.

Adoptive Treg therapy with metabolic intervention via perforated microneedles ameliorates psoriasis syndrome.

Regulatory T (Treg) cells underlie multiple autoimmune disorders and potentialize an anti-inflammation treatment with adoptive cell therapy. However, systemic delivery of cellular therapeutics often lacks tissue targeting and accumulation for localized autoimmune diseases. Besides, the instability and plasticity of Treg cells also induce phenotype transition and functional loss, impeding clinical translation. Here, we developed a perforated microneedle (PMN) with favorable mechanical performance and a spacious encapsulation cavity to support cell survival, as well as tunable channels to facilitate cell migration for local Treg therapy of psoriasis. In addition, the enzyme-degradable microneedle matrix could release fatty acid in the hyperinflammatory area of psoriasis, enhancing the Treg suppressive functions via the fatty acid oxidation (FAO)-mediated metabolic intervention. Treg cells administered through PMN substantially ameliorated psoriasis syndrome with the assistance of fatty acid-mediated metabolic intervention in a psoriasis mouse model. This tailorable PMN could offer a transformative platform for local cell therapy to treat a variety of diseases.

Engineered bacteria for augmented in situ tumor vaccination.

In situ tumor vaccination has aroused tremendous interest with its capability for eliciting strong and systemic antitumor immune responses. Unlike traditional cancer vaccines, in situ tumor vaccination avoids the laborious process of tumor antigen identification and can modulate tumor immunosuppressive microenvironment at the same time. In recent years, bacteria have been used as both efficient tumor-targeted delivery vehicles and potent adjuvants. Regarding the rapid development in this area, in this review, we summarize recent advances in the application of bacteria for in situ cancer vaccination. We illustrate the mechanisms of bacteria as both efficient tumor immunogenic cell death inducers and tumor-targeted delivery platforms. Then we comprehensively review the engineering strategies for designing bacteria-based in situ vaccination, including chemical modification, nanotechnology, and genetic engineering. The current dilemma and future directions are discussed at the end of this review.

Cell delivery devices for cancer immunotherapy.

Adoptive cell therapy (ACT) that leverages allogeneic or autologous immune cells holds vast promise in targeted cancer therapy. Despite the tremendous success of ACT in treating hematopoietic malignancies, its efficacy is limited in eradicating solid tumors via intravenous infusion of immune cells. With the extending technology of cancer immunotherapy, novel delivery strategies have been explored to improve the therapeutic potency of adoptively transferred cells for solid tumor treatment by innovating the administration route, maintaining the cell viability, and normalizing the tumor microenvironment. In this review, a variety of devices for cell delivery are summarized. Perspectives and challenges of cell delivery devices for cancer immunotherapy are also discussed.

Half-millennium evidence suggests that extinction debts of global vertebrates started in the Second Industrial Revolution.

Extinction debt describes the time-lagged process of species extinction, which usually requires dozens to hundreds of years to be paid off. However, due to the lack of long-term habitat data, it is indeterminate how strong the signal of extinction debts is at the global scale and when the debts started. Here, by compiling the geographical distributions of 6120 reptiles, 6047 amphibians, and 4278 mammals and correlating them with annual forest cover data from 1500 to 1992, we show that the beginning of the Second Industrial Revolution (the mid-19th century) was the earliest signal of cumulative extinction debts for global forest-dwelling vertebrate groups. More importantly, the impact of global protected areas on mitigating accumulated vertebrate extinction debt is not as immediate as that of mitigating reduced forest cover but rather suffers from pronounced time-lag effects. As the disequilibrium of vertebrate richness and forested habitat is currently taking place, preventive actions should be taken to promote a well-balanced status among forest restoration, protected areas, and biodiversity conservation to slow the accumulating debts for global forest-dwelling vertebrates.

Climatic Variability Caused by Topographic Barrier Prevents the Northward Spread of Invasive Ageratina adenophora.

Ageratina adenophora (Spreng.) R.M.King & H.Rob. is one of the most threatening invasive alien plants in China. Since its initial invasion into Yunnan in the 1940s, it spread rapidly northward to southern Mount Nyba in Sichuan, which lies on the eastern edge of the Qinghai-Tibet Plateau. During fieldwork, we found an interesting phenomenon: A. adenophora failed to expand northward across Mount Nyba, even after the opening of the 10 km tunnel, which could have served as a potential corridor for its spread. In this work, to explore the key factors influencing its distribution and spread patterns, we used a combination of ensemble species distribution models with the MigClim model. We found that the temperature annual range (TAR), precipitation of driest month (PDM), highway density (HW), and wind speed (WS) were the most predominant factors affecting its distribution. The north of Mount Nyba is not suitable for A. adenophora survival due to higher TAR. The spatial-temporal dynamic invasion simulation using MigClim further illustrated that the northward invasion of A. adenophora was stopped by Mount Nyba. Overall, Mount Nyba may act as a topographic barrier that causes environmental differences between its south and north sides, preventing the northward invasion of A. adenophora. However, other suitable habitats on the northern side of the mountain still face challenges because A. adenophora is likely to invade via other routes. Therefore, long-term monitoring is needed to prevent human-induced long-distance spread events.

Evaluation of the Carbon Sink Capacity of the Proposed Kunlun Mountain National Park.

National parks, as an important type of nature protected areas, are the cornerstone that can effectively maintain biodiversity and mitigate global climate change. At present, China is making every effort to build a nature-protection system, with national parks as the main body, and this approach considers China's urgent goals of obtaining carbon neutrality and mitigating climate change. It is of great significance to the national carbon-neutralization strategy to accurately predict the carbon sink capacity of national park ecosystems under the background of global change. To evaluate and predict the dynamics of the carbon sink capacity of national parks under climate change and different management measures, we combined remote-sensing observations, model simulations and scenario analyses to simulate the change in the carbon sink capacity of the proposed Kunlun Mountain National Park ecosystem over the past two decades (2000-2020) and the change in the carbon sink capacity under different zoning controls and various climate change scenarios from 2020 to 2060. Our results show that the carbon sink capacity of the proposed Kunlun Mountain National Park area is increasing. Simultaneously, the carbon sink capacity will be improved with the implementation of park management and control measures; which will be increased by 2.04% to 2.13% by 2060 in the research area under multiple climate change scenarios. The research results provide a scientific basis for the establishment and final boundary determination of the proposed Kunlun Mountain National Park.

Leveraging biomaterials for enhancing T cell immunotherapy.

The dynamic roles of T cells in the immune system to recognize and destroy the infected or mutated cells render T cell therapy a prospective treatment for a variety of diseases including cancer, autoimmune diseases, and allograft rejection. However, the clinical applications of T cell therapy remain unsatisfactory due to the tedious manufacturing process, off-target cytotoxicity, poor cell persistence, and associated adverse effects. To this end, various biomaterials have been introduced to enhance T cell therapy by facilitating proliferation, enhancing local enrichment, prolonging retention, and alleviating side effects. This review highlights the design strategies of biomaterials developed for T cell expansion, enrichment, and delivery as well as their corresponding therapeutic effects. The prospects of biomaterials for enhancing T cell immunotherapy are also discussed in this review.

Urbanization increases stochasticity and reduces the ecological stability of microbial communities in amphibian hosts.

Urbanization not only profoundly alters landscape profiles, ecosystems and vertebrate faunal diversity but also disturbs microbial communities by increasing stochasticity, vulnerability, biotic homogenization, etc. However, because of the buffering effect of host species, microbial communities are expected to be influenced by both host species and urbanization stresses. Therefore, the impacts of urbanization on animals' microbial symbionts could be more complex and uncertain. In this study, we quantified the urbanization degree of sampling sites and surveyed the gut and skin microbes of three amphibian host species in different sites in urban parks and nearby villages of Chengdu, Southwest China. Furthermore, a co-occurrence network analysis, the phylogenetic normalized stochasticity ratio and Sloan neutral community models were applied to infer the impact of urbanization on symbiotic microbial communities. For the three host species, urbanization increased the diversity of symbiotic microbes and the number of keystone microbial taxa. However, the negative effects of such increased diversification were evident, as the community stochasticity and co-occurrence network structure vulnerability also increased, while the network structure complexity and stability were reduced. Finally, the community stochasticity had positive associations with the network vulnerability, implying that the existence of many transient symbiotic rare microbial taxa in urban parks makes the symbiotic microbial community structure more fragile. Conclusively, urbanization increased the symbiotic microbial diversity at the cost of community stability; the results provide a new perspective for better understanding the complex triangulated environment-host-microbe relationship.

Species richness and endemism pattern of Fagaceae in Southwest China and their environmental interpretation.

How to accurately model species macro-richness patterns and endemism centers is a key focus of biodiversity conservation efforts and a hot biogeographical topic. Southwest China is one of regions with high Fagaceae species richness, the species diversity patterns and driving mechanisms are unclear. In this study, the distribution pattern of species richness (SR), weighted endemism (WE), and corrected weighted endemism (CWE) indices were estimated based on 7258 occurrence points of 161 Fagaceae species in Southwest China using both occurrence-to grid method and species distribution model (SDM). We used the spatial autoregressive (SAR) model to analyze the relationship between diversity indices and environmental factors. Overall, the three SDM-simulated diversity indices were more continuous in values than that of the occurrence-to grid method, though the distributions of those indices obtained by the two methods were similar. The areas with high SR value were mainly distributed in the south edge of Yunnan, north Guangxi and southwest Guangxi (62-89 species). The maximum of WE concentrated in south Yunnan and west Guangxi (1.77-5.02). The highest CWE (0.07-0.17) was found in southeast Tibet, Qinling-Daba Mountains, southwest Guangxi, and southeast Yunnan. The SAR models showed significant effect of precipita-tion in the driest month, standard deviations of seasonal temperature, altitude range and soil organic carbon content on SR. The effects of precipitation in the driest month, standard deviations of seaso-nal temperature, potential evaporation and altitude range on the WE were significant. The precipitation in the driest month, standard deviations of seasonal temperature, historical temperature change, coefficient of variation of enhanced vegetation index and altitude variation had significant effects on the CWE. The R2 of SAR model for SR, WE and CWE was 0.857, 0.733, 0.593, respectively, being higher than that of ordinary least squares (OLS) (R2=0.689, 0.425, 0.422). In conclusion, water availability, climate seasonality, habitat heterogeneity, historical climate change and soil condition were the most important factors limiting the distribution of SR and WE of Fagaceae in Southwest China. The SR and WE centers of Fagaceae were located in south and southeast Yunnan, southwest Guangxi, west Guangxi, Qinling-Daba Mountains, and southeast Tibet, where should be adequately protected.

Energy-water and seasonal variations in climate underlie the spatial distribution patterns of gymnosperm species richness in China.

Studying the pattern of species richness is crucial in understanding the diversity and distribution of organisms in the earth. Climate and human influences are the major driving factors that directly influence the large-scale distributions of plant species, including gymnosperms. Understanding how gymnosperms respond to climate, topography, and human-induced changes is useful in predicting the impacts of global change. Here, we attempt to evaluate how climatic and human-induced processes could affect the spatial richness patterns of gymnosperms in China. Initially, we divided a map of the country into grid cells of 50 × 50 km2 spatial resolution and plotted the geographical coordinate distribution occurrence of 236 native gymnosperm taxa. The gymnosperm taxa were separated into three response variables: (a) all species, (b) endemic species, and (c) nonendemic species, based on their distribution. The species richness patterns of these response variables to four predictor sets were also evaluated: (a) energy-water, (b) climatic seasonality, (c) habitat heterogeneity, and (d) human influences. We performed generalized linear models (GLMs) and variation partitioning analyses to determine the effect of predictors on spatial richness patterns. The results showed that the distribution pattern of species richness was highest in the southwestern mountainous area and Taiwan in China. We found a significant relationship between the predictor variable set and species richness pattern. Further, our findings provide evidence that climatic seasonality is the most important factor in explaining distinct fractions of variations in the species richness patterns of all studied response variables. Moreover, it was found that energy-water was the best predictor set to determine the richness pattern of all species and endemic species, while habitat heterogeneity has a better influence on nonendemic species. Therefore, we conclude that with the current climate fluctuations as a result of climate change and increasing human activities, gymnosperms might face a high risk of extinction.

Interfacing DNA with nanoparticles: Surface science and its applications in biosensing.

The knowledge on the mechanisms of DNA interfacing with nanoparticles holds great potential for the design, assembly and usage of DNA in biological applications. A wave of understanding and exploitation of the mechanisms in DNA-nanoparticles interfacial phenomenon has raised. Although some previous reviews have been reported, systematic and detailed reviews are rare. To achieve a better understanding of the mechanisms in the interaction between DNA and nanoparticles, here, we summarized the recent progresses on the fundamental principles regarding the DNA-nanoparticle interactions and their applications in biosensing. Special focus was put on inorganic nanoparticles such as metal nanoparticles, carbon-based materials, metal oxides and quantum dots. For each material, the surface properties, the interfacing mechanisms, and the kinetics and spatial control of DNA adsorption were summarized and discussed. We also highlighted some of the recent technologies based on DNA-NPs interactions for biomolecules detection. Finally, the challenges and future directions were discussed and proposed. This review provides a systematic understanding of the mechanisms in the interaction of DNA-nanoparticles, which, in turn, can inspire new insights for designing biosensors with improved properties.

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The complexity and uncertainty of forest regeneration is crucial for predicting forest ecosystem dynamics. A natural regeneration model of pine-oak forests in Qinling Mountains was constructed with competition, climate and topography factors using Bayesian statistics and global sensitivity analysis (GSA). The alternative models were based on Poisson, negative binomial (NB), zero-inflated Poisson (ZIP), and zero-inflated negative binomial (ZINB) models. According to the uncertainty of model parameter transfer, the analysis results were quantified, and the dominant factors of small probability events affecting forest regeneration were explained. The results showed that the ZINB model was the best one in the simulation of Pinus tabuliformis and Quercus aliena var. acuteserrata. Stand basal area, light interception, slope location and minimum temperature during growing season were the most critical factors affecting natural regeneration of P. tabuliformis, while stand basal area, cosine of aspect interacted with the natural logarithm of elevation, annual mean temperature, and precipitation of the warmest quarter were the most critical factors for Q. aliena var. acuteserrata. The contributions of various factors to the predictive uncertainty were: competition factor (25%) < climate factor (29%) < topography factor (46%) for the simulation of P. tabuliformis regeneration, and climate factor (12%) < competition factor (24%) < topography factor (64%) for the simulation of Q. aliena var. acuteserrata regeneration. The natural regeneration quantity of P. tabuliformis was positively correlated with mean annual temperature and minimum precipitation during growing season, and negatively correlated with the mean temperature in the driest quarter. The natural regeneration quantity of Q. aliena var. acuteserrata was positively correlated with mean annual temperature, minimum precipitation during growing season, precipitation of the warmest quarter, and negatively correlated with mean temperature of the driest quarter. The ZINB model based on Bayesian methods could effectively quantify the major factors driving forest regeneration and interpret the uncertainty propagated from parameters, which was useful for predicting forest regeneration.