DNA Polymerase Delta Synthesizes Both Strands during Break-Induced Replication.

Break-induced replication (BIR) is a pathway of homology-directed repair that repairs one-ended DNA breaks, such as those formed at broken replication forks or uncapped telomeres. In contrast to conventional S phase DNA synthesis, BIR proceeds by a migrating D-loop and results in conservative synthesis of the nascent strands. DNA polymerase delta (Pol δ) initiates BIR; however, it is not known whether synthesis of the invading strand switches to a different polymerase or how the complementary strand is synthesized. By using alleles of the replicative DNA polymerases that are permissive for ribonucleotide incorporation, thus generating a signature of their action in the genome that can be identified by hydrolytic end sequencing, we show that Pol δ replicates both the invading and the complementary strand during BIR. In support of this conclusion, we show that depletion of Pol δ from cells reduces BIR, whereas depletion of Pol ε has no effect.

Herbivore-induced volatiles reduce the susceptibility of neighboring tomato plants to transmission of a whitefly-borne begomovirus.

Plant viruses exist in a broader ecological community, with key components include non-vector herbivores that can impact vector abundance, behavior, and virus transmission within shared host plants. However, little is known about the effects of non-vector herbivores infestation on the virus transmission by vector insects on the neighboring plants through inter-plant airborne chemicals. In this study, we investigated how volatiles emitted from tomato plants infested with the two-spotted spider mite (Tetranychus urticae) affect the infection of Tomato yellow leaf curl virus (TYLCV) transmitted by the whitefly (Bemisia tabaci) in the neighboring plants. Exposure of neighboring tomato plants to volatiles released from T. urticae-infested tomato plants reduced subsequent herbivory as well as TYLCV transmission and infection, and JA signaling pathway was essential for generation of the inter-plant defense signals. We also demonstrated that (E)-ß-Ocimene and MeSA were two volatiles induced by T. urticae that synergistically attenuated TYLCV transmission and infection in tomato. Thus, our findings suggest that plant-plant communication via volatiles likely represents a widespread defensive mechanism that substantially contributes to plant fitness. Understanding such phenomena may help us to predict the occurrence and epidemic of multiple herbivores and viruses in the agroecosystem, ultimately to manage pest and virus outbreaks.

Pif1 family helicases promote mutation avoidance during DNA replication.

Pif1 family 5' → 3' DNA helicases are important for replication fork progression and genome stability. The budding yeast Saccharomyces cerevisiae encodes two Pif1 family helicases, Rrm3 and Pif1, both of which are multi-functional. Here we describe novel functions for Rrm3 in promoting mutation avoidance during DNA replication. We show that loss of RRM3 results in elevated spontaneous mutations made by DNA polymerases Pols ϵ and δ, which are subject to DNA mismatch repair. The absence of RRM3 also causes higher mutagenesis by the fourth B-family DNA polymerase Pol ζ. By genome-wide analysis, we show that the mutational consequences due to loss of RRM3 vary depending on the genomic locus. Rrm3 promotes the accuracy of DNA replication by Pols ϵ and δ across the genome, and it is particularly important for preventing Pol ζ-dependent mutagenesis at tRNA genes. In addition, mutation avoidance by Rrm3 depends on its helicase activity, and Pif1 serves as a backup for Rrm3 in suppressing mutagenesis. We present evidence that the sole human Pif1 family helicase in human cells likely also promotes replication fidelity, suggesting that a role for Pif1 family helicases in mutation avoidance may be evolutionarily conserved, a possible underlying mechanism for its potential tumor-suppressor function.

Association of physical activity with socio-economic status and chronic disease in older adults in China: cross-sectional findings from the survey of CLASS 2020 after the outbreak of COVID-19.

BACKGROUND: In 2021, China had a population of 264·01 million individuals over the age of 60, indicating a high prevalence of chronic diseases. Among older adults, physical inactivity (PI) is a significant risk factor for chronic diseases. However, few studies have been conducted on the correlation of physical activity (PA) with the economic status, geography and chronic disease risks in Chinese elderly. The objectives of this study were to better understand the distribution of PA among older adults in China and its relationship with economic status, geography, and chronic disease risks. METHODS: This study utilized data from the China Longitudinal Aging Social Survey (CLASS) in 2020, post-COVID-19. The study employed a stratified, multistage, probabilistic sampling approach and included 11,396 adults over the age of 59 from 28 provinces in China. Data on demographics, the duration and intensity of PA, history of diseases and personalized factors influencing PA were collected via structured interviews by researchers. In this study, we conducted a comprehensive analysis, employing a range of statistical methods including descriptive analysis, Wilcoxon rank-sum tests, Bayesian networks, and chi-square tests. RESULTS: The prevalence of PI among older adults over 59 in China is 28·82%. Significant regional differences were observed in the duration of PA at different intensities. Older adults residing in more economically developed areas were more likely to engage in moderate-to-vigorous physical activity (MVPA) and exhibited longer sedentary behavior. Economic status and urban-rural disparities consistently emerged as direct influential factors across all intensity types. Chronic disease risks were significantly lower in active older adults compared to inactive ones. Lack of social guidance, family support, and personal inclination towards sedentary behavior were the main personalized factors affecting PA among older adults, and these factors could be relatively easily modified. CONCLUSIONS: Economic status, geography, and living areas (urban and rural) significantly influenced the distribution of physical activities in China. Particularly, economic status and living areas acted as direct factors. Older adults reaching the recommended standards for PA had significantly lower chronic disease risks, highlighting the importance of improving personalized factors which are crucial for promoting PA.

An Effective Deep Learning Framework for Fall Detection: Model Development and Study Design.

BACKGROUND: Fall detection is of great significance in safeguarding human health. By monitoring the motion data, a fall detection system (FDS) can detect a fall accident. Recently, wearable sensors-based FDSs have become the mainstream of research, which can be categorized into threshold-based FDSs using experience, machine learning-based FDSs using manual feature extraction, and deep learning (DL)-based FDSs using automatic feature extraction. However, most FDSs focus on the global information of sensor data, neglecting the fact that different segments of the data contribute variably to fall detection. This shortcoming makes it challenging for FDSs to accurately distinguish between similar human motion patterns of actual falls and fall-like actions, leading to a decrease in detection accuracy. OBJECTIVE: This study aims to develop and validate a DL framework to accurately detect falls using acceleration and gyroscope data from wearable sensors. We aim to explore the essential contributing features extracted from sensor data to distinguish falls from activities of daily life. The significance of this study lies in reforming the FDS by designing a weighted feature representation using DL methods to effectively differentiate between fall events and fall-like activities. METHODS: Based on the 3-axis acceleration and gyroscope data, we proposed a new DL architecture, the dual-stream convolutional neural network self-attention (DSCS) model. Unlike previous studies, the used architecture can extract global feature information from acceleration and gyroscope data. Additionally, we incorporated a self-attention module to assign different weights to the original feature vector, enabling the model to learn the contribution effect of the sensor data and enhance classification accuracy. The proposed model was trained and tested on 2 public data sets: SisFall and MobiFall. In addition, 10 participants were recruited to carry out practical validation of the DSCS model. A total of 1700 trials were performed to test the generalization ability of the model. RESULTS: The fall detection accuracy of the DSCS model was 99.32% (recall=99.15%; precision=98.58%) and 99.65% (recall=100%; precision=98.39%) on the test sets of SisFall and MobiFall, respectively. In the ablation experiment, we compared the DSCS model with state-of-the-art machine learning and DL models. On the SisFall data set, the DSCS model achieved the second-best accuracy; on the MobiFall data set, the DSCS model achieved the best accuracy, recall, and precision. In practical validation, the accuracy of the DSCS model was 96.41% (recall=95.12%; specificity=97.55%). CONCLUSIONS: This study demonstrates that the DSCS model can significantly improve the accuracy of fall detection on 2 publicly available data sets and performs robustly in practical validation.

Ribonucleotide incorporation into DNA during DNA replication and its consequences.

Ribonucleotides are the most abundant non-canonical nucleotides in the genome. Their vast presence and influence over genome biology is becoming increasingly appreciated. Here we review the recent progress made in understanding their genomic presence, incorporation characteristics and usefulness as biomarkers for polymerase enzymology. We also discuss ribonucleotide processing, the genetic consequences of unrepaired ribonucleotides in DNA and evidence supporting the significance of their transient presence in the nuclear genome.

Strand-specific analysis shows protein binding at replication forks and PCNA unloading from lagging strands when forks stall.

In eukaryotic cells, DNA replication proceeds with continuous synthesis of leading-strand DNA and discontinuous synthesis of lagging-strand DNA. Here we describe a method, eSPAN (enrichment and sequencing of protein-associated nascent DNA), which reveals the genome-wide association of proteins with leading and lagging strands of DNA replication forks. Using this approach in budding yeast, we confirm the strand specificities of DNA polymerases delta and epsilon and show that the PCNA clamp is enriched at lagging strands compared with leading-strand replication. Surprisingly, at stalled forks, PCNA is unloaded specifically from lagging strands. PCNA unloading depends on the Elg1-containing alternative RFC complex, ubiquitination of PCNA, and the checkpoint kinases Mec1 and Rad53. Cells deficient in PCNA unloading exhibit increased chromosome breaks. Our studies provide a tool for studying replication-related processes and reveal a mechanism whereby checkpoint kinases regulate strand-specific unloading of PCNA from stalled replication forks to maintain genome stability.

Evaluating the Effects of Cu2+ on the Development and Reproduction of Spodoptera litura (Lepidoptera: Noctuidae) Based on the Age-Stage, Two-Sex Life Table.

Copper (Cu2+) is a micronutrient that promotes the development and reproduction of organisms. However, with the rapid expansion of modern industry and agriculture, Cu2+ concentrations are increasing, which might have negative impacts on biological and ecological safety. Spodoptera litura is not only an intermittent outbreak pest but also can be used as a model organism to assess environmental and ecological risks. Therefore, the effects of the life history and population parameters of S. litura fed on artificial diets with different Cu2+ concentrations were analyzed using the age-stage, two-sex life table. Our results showed that not only the preadult survival rate but also the intrinsic rate of increase (r) and the finite rate of increase (λ) were significantly increased under exposure to low Cu2+ concentrations (2, 4, and 8 mg/kg). In addition, the population growth of S. litura was significantly faster, indicating that S. litura can adapt well to low concentrations and is likely to undergo outbreaks of damage. Whereas, in addition to a significant reduction in preadult survival rate, population growth rate, pupal weight, pupal length, adult body weight, and oviposition were also significantly reduced under exposure to high Cu2+ concentration (32 mg/kg). And when the concentration reached 64 mg/kg, the survival rate of adults was extremely low, suggesting a decrease in the adaptation of S. litura. These results can help to understand the population dynamics of S. litura and predict potential ecological risks.

Fabrication of Fresnel lens arrays by a rapid non-isothermal imprinting process.

Fresnel lens arrays are widely employed in concentrator photovoltaics, photonic devices, and integral imaging systems. In this study, a rapid non-isothermal imprinting process for Fresnel lens arrays was proposed. In this process, a heated mold with microstructures was momentarily pressed onto a thermoplastic polymer surface that was initially kept at room temperature. The microstructures of the mold can be copied completely to the polymer substrate by imprinting consecutively until a continuous surface Fresnel lens array is obtained. Different from more traditional molding processes, the substrate does not need to be heated and cooled repeatedly in the replicating process. In addition, the imprinting process is carried out at room temperature, which can greatly reduce the thermal cycle time and energy consumption. Generally speaking, the material flow and stress distribution of the substrate need to be monitored so that the microlenses with a high precision surface finish can be produced in the non-isothermal imprinting process. To verify this, the finite element method (FEM) model for the non-isothermal process was established, and the feasibility of this process was analyzed. A hexagonal continuous surface Fresnel lens array was then fabricated, and its geometrical contour and imaging performance were tested. The experimental results showed this new process could be an effective and low-cost optical fabrication technology for high-quality production of Fresnel lens arrays.

The Influence of Gut Microbiota on the Fecundity of Henosepilachna vigintioctopunctata (Coleoptera: Coccinellidae).

The gut microbiota of insects usually plays an important role in the development and reproduction of their hosts. The fecundity of Henosepilachna vigintioctopunctata (Fabricius) varies greatly when they develop on different host plants. Whether and how the gut microbiota regulates the fecundity of H. vigintioctopunctata was unknown. To address this question, we used 16S rRNA sequencing to analyze the gut microbiomes of H. vigintioctopunctata adults fed on two host plant species (Solanum nigrum and Solanum melongena) and one artificial diet. The development of the ovaries and testes was also examined. Our results revealed that the diversity and abundance of gut microorganisms varied significantly in insects reared on different diets. The gut microbiota of H. vigintioctopunctata raised on the two host plants was similar, with Proteobacteria being the dominant phylum in both groups, whereas Firmicutes was the dominant phylum in the group reared on the artificial diet. The predominant microbiota in the S. nigrum group were Acinetobacter soli and Acinetobacter ursingii (Acinetobacter, Moraxellaceae); Moraxella osloensis (Enhydrobacter, Moraxellaceae); and Empedobacter brevis (Empedobacter, Weeksellaceae). The microbiota in this group are associated with high lipid metabolism. In addition, the beetles' ovaries and testes were more highly developed in the S. nigrum group than in the other two groups. These findings provide valuable information for elucidating the complex roles the gut microbiota play in the fecundity of H. vigintioctopunctata, and may also contribute to developing future novel control strategies involving this economically important pest.

The Mechanism of Facultative Intracellular Parasitism of Brucella.

Brucellosis is a highly prevalent zoonotic disease characterized by abortion and reproductive dysfunction in pregnant animals. Although the mortality rate of Brucellosis is low, it is harmful to human health, and also seriously affects the development of animal husbandry, tourism and international trade. Brucellosis is caused by Brucella, which is a facultative intracellular parasitic bacteria. It mainly forms Brucella-containing vacuoles (BCV) in the host cell to avoid the combination with lysosome (Lys), so as to avoid the elimination of it by the host immune system. Brucella not only has the ability to resist the phagocytic bactericidal effect, but also can make the host cells form a microenvironment which is conducive to its survival, reproduction and replication, and survive in the host cells for a long time, which eventually leads to the formation of chronic persistent infection. Brucella can proliferate and replicate in cells, evade host immune response and induce persistent infection, which are difficult problems in the treatment and prevention of Brucellosis. Therefore, the paper provides a preliminary overview of the facultative intracellular parasitic and immune escape mechanisms of Brucella, which provides a theoretical basis for the later study on the pathogenesis of Brucella.

The VirB System Plays a Crucial Role in Brucella Intracellular Infection.

Brucellosis is a highly prevalent zoonotic disease caused by Brucella. Brucella spp. are gram-negative facultative intracellular parasitic bacteria. Its intracellular survival and replication depend on a functional virB system, an operon encoded by VirB1-VirB12. Type IV secretion system (T4SS) encoded by the virB operon is an important virulence factor of Brucella. It can subvert cellular pathway and induce host immune response by secreting effectors, which promotes Brucella replication in host cells and induce persistent infection. Therefore, this paper summarizes the function and significance of the VirB system, focusing on the structure of the VirB system where VirB T4SS mediates biogenesis of the endoplasmic reticulum (ER)-derived replicative Brucella-containing vacuole (rBCV), the effectors of T4SS and the cellular pathways it subverts, which will help better understand the pathogenic mechanism of Brucella and provide new ideas for clinical vaccine research and development.

Comparison of canal transportation and centering ability of manual K-files and reciprocating files in glide path preparation: a micro-computed tomography study of constricted canals.

BACKGROUND: Optimum Glide Path (OGP) is a new reciprocating motion aiming to perform efficient glide path preparation in constricted canals. The aim of this study was to investigate and compare manual and OGP movement in terms of canal transportation and centering ability in glide path preparation of constricted canals. METHODS: Thirty constricted mesial root canals of mandibular molars, with initial apical size no larger than ISO#8, were selected and negotiated with #6-#8 K-files under the microscope. Canals were randomly divided into two experimental groups: Group 1 (MAN, n = 15): Glide path was established by using #10-#15 stainless steel K-files manually; Group 2 (OGP, n = 15): #10-#15 Mechanical Glide Path super-files were used with OGP motion (OGP 90°, 300 rpm). Each instrument was used to prepare only 2 canals (as in one mesial root). Canals were scanned before and after glide path preparation with micro-computed tomography (micro-CT) to evaluate root canal transportation and centering ratio at 1, 3 and 5 mm levels from the root apex. File distortions and separations were recorded. Paired t-test was used to statistically evaluate the data (P < .05). RESULTS: Group 2 showed a significantly lower transportation value than group 1 at 1-mm and 3-mm levels (P < .05), however the difference at 5-mm level was not significant. There was no significant difference regarding the centering ratio between the groups. Six #10 K-files were severely distorted in group 1, while no file separation or distortion was found in group 2. CONCLUSIONS: OGP motion performed significantly less canal transportation (apical 3 mm) and file distortion during glide path establishment in constricted canals comparing to manual motion, while the centering ability between the two was similar. CLINICAL RELEVANCE: OGP reciprocating motion provides a safer and efficient clinical approach compared to traditional manual motion in glide path establishment with small files in constricted canals.

Effects of Pre-Diapause Temperature and Body Weight on the Diapause Intensity of the Overwintering Generation of Bactrocera minax (Diptera: Tephritidae).

The Chinese citrus fruit fly, Bactrocera minax (Enderlein), is an economically important pest of citrus. The fly has an obligatory pupal diapause in soil from November to March. However, techniques for predicting or determining the emergence of the adult have, thus far, not been well documented. In this study, we investigated the effects of different pre-diapause temperatures (8, 12, 16, and 20°C) and pupal body weight (five groups according to pupal weight: G-58, 55.0-61.0 mg; G-68, 65-71 mg; G-78, 75-81 mg; G-88, 85-91 mg; G-95, 92-98 mg) on pupal period (the indicator of diapause intensity). The pupal period of B. minax larvae pupated at 8°C was 193.41 d, which was significantly shorter than that of larvae incubated at higher temperatures, suggesting that there was a lower diapause intensity for larvae pupated at lower pre-diapause temperatures. There were also significant differences in the pupal periods at different pupal body weights. The pupal period of G-58 was significantly shorter than that of the heavier groups (G-88 and G-95), and the pupal period increased with increasing pupal body weight in the five groups. Moreover, the pupal period of B. minax significantly and positively correlated to pupal body weight. These findings demonstrate that the pre-diapause temperature and pupal body weight are suitable indicators for predicting the pupal period of overwintering individuals, and the results of this study will contribute to the development of new and effective strategies for predicting the occurrence and population dynamics of B. minax adult.

Chemical Composition and Attractant Activity of Volatiles from Rhus potaninii to The Spring Aphid Kaburagia rhusicola.

Rhus potaninii Maxim, a type of sumac, is an economically important tree widely cultivated in mountainous areas of western and central China. A gall, called the bellied gallnut, induced by the aphid, Kaburagia rhusicola Takagi, is important in the food, medical, and chemical industries in China. Volatiles from R. potaninii were found to attract K. rhusicola, but little is known about them. The chemical composition of these volatiles was investigated using GC-MS analysis and Y-tube olfactometer methods. Twenty-five compounds accounting for 55.3% of the volatiles were identified, with the highest proportion of 1-(4-ethylphenyl)ethanone (11.8%), followed by 1-(4-hydroxy-3-methylphenyl)ethanone (11.2%) and p-cymen-7-ol (7.1%). These findings provide a theoretical basis for the preparation of attractants and could eventually lead to increased bellied gallnut yield.

Fission yeast Pxd1 promotes proper DNA repair by activating Rad16XPF and inhibiting Dna2.

Structure-specific nucleases play crucial roles in many DNA repair pathways. They must be precisely controlled to ensure optimal repair outcomes; however, mechanisms of their regulation are not fully understood. Here, we report a fission yeast protein, Pxd1, that binds to and regulates two structure-specific nucleases: Rad16XPF-Swi10ERCC1 and Dna2-Cdc24. Strikingly, Pxd1 influences the activities of these two nucleases in opposite ways: It activates the 3' endonuclease activity of Rad16-Swi10 but inhibits the RPA-mediated activation of the 5' endonuclease activity of Dna2. Pxd1 is required for Rad16-Swi10 to function in single-strand annealing, mating-type switching, and the removal of Top1-DNA adducts. Meanwhile, Pxd1 attenuates DNA end resection mediated by the Rqh1-Dna2 pathway. Disabling the Dna2-inhibitory activity of Pxd1 results in enhanced use of a break-distal repeat sequence in single-strand annealing and a greater loss of genetic information. We propose that Pxd1 promotes proper DNA repair by differentially regulating two structure-specific nucleases.

Excessive homework, inadequate sleep, physical inactivity and screen viewing time are major contributors to high paediatric obesity.

AIM: This study examined the relationships between energy balance-related behaviours (EBRBs) outside school hours and obesity in Chinese primary school students. We also explored the influence of gender on those relationships. METHODS: The study sample was a cross-sectional cohort of 5032 Chinese children who were enrolled in grades 1-6 in primary schools in five Chinese cities and whose mean ages ranged from seven years and three months to 11.9 years. The children's parents completed a survey on their child's height, weight and EBRBs outside school hours. RESULTS: The response rate was 97%, and the reported rates of overweight and obesity were 13.6% and 13.8%, respectively. The obesity rates were higher in boys and lower grade children. Most EBRBs varied between boys and girls and with increased grade levels. The amount of time spent on academic-related activities, screen viewing, outdoor activities and sleep was mostly associated with obesity on weekdays and varied by gender. CONCLUSION: Rate of obesity was alarmingly high in the primary school Chinese children in this cohort, especially in younger children. Excessive time spent on academic-related activities outside school hours, inadequate sleep, physical inactivity and higher levels of screen viewing were major contributors to obesity in these Chinese children.

Mapping genomic hotspots of DNA damage by a single-strand-DNA-compatible and strand-specific ChIP-seq method.

Spontaneous DNA damage may occur nonrandomly in the genome, especially when genome maintenance mechanisms are undermined. We developed single-strand DNA (ssDNA)-associated protein immunoprecipitation followed by sequencing (SPI-seq) to map genomic hotspots of DNA damage. We demonstrated this method with Rad52, a homologous recombination repair protein, which binds to ssDNA formed at DNA lesions. SPI-seq faithfully detected, in fission yeast, Rad52 enrichment at artificially induced double-strand breaks (DSBs) as well as endogenously programmed DSBs for mating-type switching. Applying Rad52 SPI-seq to fission yeast mutants defective in DNA helicase Pfh1 or histone H3K56 deacetylase Hst4, led to global views of DNA lesion hotspots emerging in these mutants. We also found serendipitously that histone dosage aberration can activate retrotransposon Tf2 and cause the accumulation of a Tf2 cDNA species bound by Rad52. SPI-seq should be widely applicable for mapping sites of DNA damage and uncovering the causes of genome instability.

Longer rest intervals do not attenuate the superior effects of accumulated exercise on arterial stiffness.

PURPOSE: To examine and compare the acute effects of moderate-intensity continuous and accumulated exercise in three bouts with different intervals on arterial stiffness. METHOD: Nineteen healthy young males (mean age = 24.7 years) were randomized to no-exercise control (CON), continuous exercise (CE, 30-min cycling), accumulated exercise with 10-min intervals (AE10, 3 × 10-min cycling, 10-min interval), and accumulated exercise with 60-min intervals (AE60, 3 × 10-min cycling, 60-min interval) trial in balanced self-control crossover design. The intensity in all the exercise trials was set at 50% heart rate reserve. Cardio-ankle vascular index (CAVI), an index of arterial stiffness, was measured at baseline (BL), immediately after (0 min) and 60 min after the completion of the exercise. RESULTS: CAVI remained stable (6.8 ± 0.1, 6.8 ± 0.2, 6.9 ± 0.1 at BL, 0 and 60 min, respectively) in CON trial. Immediately after exercise, CAVI in CE, AE10 and AE60 trials all decreased significantly to similar degree compared to CON trial (P < 0.05 for CE, AE10 and AE60 vs. CON). Though CAVI in CE trial returned to baseline level after 60 min of recovery, CAVI in both AE10 and AE60 trials remained significantly low compared to CON trial (P < 0.01 for AE10 and AE60 vs. CON). CONCLUSION: When the total duration and relative intensity were matched, the effects of accumulated exercise in three bouts were superior to continuous exercise. Elongation of intervals between bouts did not attenuate the superior effects of accumulated exercise on arterial stiffness. TRIAL REGISTRATION: ChiCTR-OTRCC-14005229.

A policy-driven multifaceted approach for early childhood physical fitness promotion: impacts on body composition and physical fitness in young Chinese children.

BACKGROUND: The prevalence of obesity increased while certain measures of physical fitness deteriorated in preschool children in China over the past decade. This study tested the effectiveness of a multifaceted intervention that integrated childcare center, families, and community to promote healthy growth and physical fitness in preschool Chinese children. METHODS: This 12-month study was conducted using a quasi-experimental pretest/posttest design with comparison group. The participants were 357 children (mean age = 4.5 year) enrolled in three grade levels in two childcare centers in Beijing, China. The intervention included: 1) childcare center intervention (physical activity policy changes, teacher training, physical education curriculum and food services training), 2) family intervention (parent education, internet website for support, and family events), and 3) community intervention (playground renovation and community health promotion events). The study outcome measures included body composition (percent body fat, fat mass, and muscle mass), Body Mass Index (BMI) and BMI z-score and physical fitness scores in 20-meter agility run (20M-AR), broad jump for distance (BJ), timed 10-jumps, tennis ball throwing (TBT), sit and reach (SR), balance beam walk (BBW), 20-meter crawl (20M-C)), 30-meter sprint (30M-S)) from a norm referenced test. Measures of process evaluation included monitoring of children's physical activity (activity time and intensity) and food preparation records, and fidelity of intervention protocol implementation. RESULTS: Children in the intervention center significantly lowered their body fat percent (-1.2%, p < 0.0001), fat mass (-0.55 kg, p <0.0001), and body weight (0.36 kg, p <0.02) and increased muscle mass (0.48 kg, p <0.0001), compared to children in the control center. They also improved all measures of physical fitness except timed 10-jumps (20M-AR: -0.74 seconds, p < 0.0001; BJ: 8.09 cm, p < 0.0001; TBT: 0.52 meters, p < 0.006; SR: 0.88 cm, p < 0.03; BBW: -2.02 seconds, p <0.0001; 30M-S: -0.45 seconds, p < 0.02; 20M-C: -3.36 seconds, p < 0.0001). Process evaluation data showed that the intervention protocol was implemented with high fidelity. CONCLUSIONS: The study demonstrated that a policy-driven multi-faceted intervention can improve preschool children's body composition and physical fitness. Program efficacy should be tested in a randomized trial. TRIAL REGISTRATION: ChiCTR-ONRC-14004143.