Revisiting hemodynamics and blood oxygenation in a microfluidic microvasculature replica.

The complexity of microvascular circulation has led to the development of advanced imaging techniques and biomimetic models. This study developed a multifaceted microfluidic-based microdevice as an in vitro model of microvasculature to replicate important geometric and functional features of in vivo perfusion in mice. The microfluidic device consisted of a microchannel for blood perfusion, mirroring the natural hierarchical branching vascular structures found in mice. Additionally, the device incorporated a steady gradient of oxygen (O2) which diffused through the polydimethylsiloxane (PDMS) layer, allowing for dynamic blood oxygenation. The assembled multi-layered microdevice was accompanied by a dual-modal imaging system that combined laser speckle contrast imaging (LSCI) and intrinsic signal optical imaging (ISOI) to visualize full-field blood flow distributions and blood O2 profiles. By closely reproducing in vivo blood perfusion and oxygenation conditions, this microvasculature model, in conjunction with numerical simulation results, can provide quantitative information on physiologically relevant hemodynamics and key O2 transport parameters that are not directly measurable in traditional animal studies.

Microplastics shift impacts of climate change on a plant-microbe mutualism: Temperature, CO2, and tire wear particles.

Anthropogenic stressors can affect individual species and alter species interactions. Moreover, species interactions or the presence of multiple stressors can modify the stressor effects, yet most work focuses on single stressors and single species. Plant-microbe interactions are a class of species interactions on which ecosystems and agricultural systems depend, yet may be affected by multiple global change stressors. Here, we use duckweed and microbes from its microbiome to model responses of interacting plants and microbes to multiple stressors: climate change and tire wear particles. Climate change is occurring globally, and microplastic tire wear particles from roads now reach many ecosystems. We paired perpendicular gradients of temperature and carbon dioxide (CO2) treatments with factorial manipulation of leachate from tire wear particles and duckweed microbiomes. We found that tire leachate and warmer temperatures enhanced duckweed and microbial growth, but caused effects of microbes on duckweed to become negative. However, induced negative effects of microbes were less than additive with warming and leachate. Without tire leachate, we observed that higher CO2 and temperature induced positive correlations between duckweed and microbial growth, which can strengthen mutualisms. In contrast, with tire leachate, growth correlations were never positive, and shifted negative at lower CO2, again suggesting leachate disrupts this plant-microbiome mutualism. In summary, our results demonstrate that multiple interacting stressors can affect multiple interacting species, and that leachate from tire wear particles could potentially disrupt plant-microbe mutualisms.

Temperature control and intermittent time-set protocol optimization for minimizing tissue carbonization in microwave ablation.

PURPOSE: The charring tissue formation in the ablated lesion during the microwave ablation (MWA) of tumors would induce various unwanted inflammatory responses. This paper aimed to deliver appropriate thermal dose for effective ablations while preventing tissue carbonization by optimizing the treatment protocol during MWA with the set combinations of temperature control and pulsed microwave energy delivery. MATERIAL AND METHODS: The thermal phase transition of ex vivo porcine liver tissues were recorded by differential scanning calorimetry (DSC) to determine the temperature threshold during microwave output control. MWA was performed by an in-house built system with the ease of microwave output parameter adjustment and real-time temperature monitoring. The effects of continuous and pulsed microwave deliveries as well as various intermittent time-set of MWA were evaluated by measuring the dimensions of the coagulation zone and the carbonization zone. RESULTS: The DSC scans demonstrated that the ex vivo porcine liver tissues have been in a state of endothermic heat during the heating process, where the maximum absorbed heat occurred at the temperature of 105 °C ± 5 °C. The temperature control during MWA resulted in effective coagulative necrosis while preventing tissue carbonization, after setting 100 °C as the upper threshold temperature and 60 °C as the lower threshold. Both the numerical simulation and ex vivo experiments have shown that, upon the optimization of the time-set parameters in the periodic intermittent pulsed microwave output, the tissue carbonization was significantly diminished. CONCLUSION: This study developed a straight-forward anti-carbonization strategy in MWA by modulating the pulsing mode and intermittent time. The programmed protocols of intermittent pulsing MWA have demonstrated its potentials toward future expansion of MWA technology in clinical application.

Image-guided simulation in comparison with laser speckle contrast imaging for full-field observation of blood flow in a microvasculature model.

The in vitro reconstruction of the microvascular network model provides a reproducible platform for hemodynamic study with great biological relevance. In the present study, microvascular models with different parametric features were designed under the guidance of Murray's law and derived from representative natural vascular network topography in vivo. Computational fluid dynamics (CFD) was used to numerically simulate blood velocity distributions inside of the designed microvasculature models. Full-field blood flow in the vascular network was visualized in vivo using a laser speckle contrast imaging (LSCI) system, from which the measured relative velocity was compared with CFD computed flow distribution. The results have shown that, in comparison with the simplified flow patterns obtained from idealized geometries, the irregular vascular topography is expected to lead to nonuniform and poor regional blood velocity distribution. The velocity distribution acquired by in vivo LSCI experiment is in good agreement with that of numerical simulation, indicating the technical feasibility of using biomimetic microchannels as a reasonable approximation of the microcirculatory flow conditions. This study provides a new paradigm that can be well suited to the study of microvascular blood flow properties and can further expand to mimic other in-vivo scenarios for accurately recapitulating the physical and hemodynamic environment of the microcirculation.

Microfluidic-enabled self-organized tumor model for in vitro cytotoxicity assessment of doxorubicin.

The advent of microfluidic technologies has enabled a better recapitulation of in vitro tumor model with higher biological relevance over conventional monolayer assays. This work built upon a microfluidic system that supported the spontaneous aggregate formation of tumoral cells under flow-induced dynamic physical forces in a confined microchamber without additional matrix materials. Our findings indicated that fluidic streams significantly modulated the biological and architectural features of human breast adenocarcinoma cell (MCF-7), human hepatocarcinoma cell (HepG2), and human cervix adenocarcinoma cell (HeLa) with cell-type-dependent variation. The microfluidic platform was further integrated with a fluorescence detection and imaging system, allowing for non-invasive monitoring of cellular accumulation and spatial distribution of a chemotherapeutic agent, doxorubicin (DOX). The cytotoxic effects of DOX of various concentrations were determined and compared in MCF-7 cells in conventional two-dimensional (2D) static and microfluidic culture conditions. Dose-dependent response to DOX was noticed in both cultures, whereas tumor micronodules grown in microfluidic devices demonstrated significantly lower sensitivity to DOX at increased concentration. Our platform owns promising potentials as a universal modality for bridging traditional 2D cell cultures and in vivo experimentation for preclinical anticancer drug screening.

Biological Responses to Climate Change and Nanoplastics Are Altered in Concert: Full-Factor Screening Reveals Effects of Multiple Stressors on Primary Producers.

While the combined presence of global climate change and nanosized plastic particle (i.e., nanoplastic) pollution is clear, the potential for interactions between climate-change-shifting environmental parameters and nanoplastics is largely unknown. Here, we aim to understand how nanoplastics will affect species in concert with climate change in freshwater ecosystems. We utilized a high-throughput full-factorial experimental system and the model photosynthetic microorganism Scenedesmus obliquus to capture the complexity of interacting environmental stressors, including CO2, temperature, light, and nanoplastics. Under a massive number of conditions (2000+), we consistently found concentration-dependent inhibition of algal growth in the presence of polystyrene nanoparticles, highlighting a threat to primary productivity in aquatic ecosystems. Our high-treatment experiment also identified crucial interactions between nanoplastics and climate change. We found that relatively low temperature and ambient CO2 exacerbated damage induced by nanoplastics, while elevated CO2 and warmer temperatures reflecting climate change scenarios somewhat attenuated nanoplastic toxicity. Further, we revealed that nanoplastics may modulate light responses, implying that risks of nanoplastic pollution may also depend on local irradiation conditions. Our study highlights the coupled impacts of nanoplastics and climate change, as well as the value of full-factorial screening in predicting biological responses to multifaceted global change.

Effects of disparity on visual discomfort caused by short-term stereoscopic viewing based on electroencephalograph analysis.

BACKGROUND: Discomfort evoked by stereoscopic depth has been widely concerned. Previous studies have proposed a comfortable disparity range and considered that disparities exceed this range would cause visual discomfort. Brain activity recordings including Electroencephalograph (EEG) monitoring enable better understanding of perceptual and cognitive processes related to stereo depth-induced visual comfort. METHODS: EEG data was collected using a stereo-visual evoked potential (VEP) test system by providing visual stimulus to subjects aged from 21 to 25 with normal stereoscopic vision. For each type of visual stimulus, data were processed using directed transfer function (DTF) and adaptive directed transfer function (ADTF) in combination with subjective feedbacks (comfort or discomfort). The topographies of information flow were constructed to compare responses stimulated by different stereoscopic depth, and to determine the difference in comfort and discomfort situations upon stimulation with same stereoscopic depth. RESULTS: Based on EEG analysis results, we found that the occipital P270 was moderately related to the disparity. Moreover, the ADTF of P270 showed that the information flows at frontal lobe and central-parietal lobe changed when stimulation with different stereoscopic depth applied. As to the stereo images with same stereoscopic depth, the DTF outflows at the temporal and temporal-parietal lobes in δ band, central and central-parietal lobes in α and θ bands, and the comparison of inflows in these three bands could be considered as discriminated indexes for matching the stereoscopic effect with viewers' comfort or discomfort state impacted by disparity. The subjective feedbacks indicated that the comfort judgments remained as a result of cumulative effect. CONCLUSIONS: This study proposed a short-term stereo-VEP experiment that shorted the duration of each stimulus in the experimental scheme to minimize the interference from other factors except the disparity. The occipital P270 had a mid-relevance to the disparity and its ADTF showed the affected areas when viewers are receiving stimulations with different disparities. DTF could be considered as discriminated indexes for matching the stereoscopic effect with viewers' comfort or discomfort state induced by disparity. This study proposed a preferable experiment to observe the single effect of disparity and provided an intuitive and easy-to-read result in a more convenient manner.

Effect of Acute Ethanol Administration on the Hippocampal Region Neural Activity Using a Microelectrode Array.

BACKGROUND: Because acute ethanol (EtOH) administration is known to influence cognitive processes by impairing hippocampal function, electrophysiological responses of the hippocampus following EtOH exposure warrant investigation. To mimic in vivo conditions, we recorded and analyzed critical firing characteristics of the neuronal population dynamically, particularly in the hippocampal region, before and after acute EtOH administration. METHODS: Microelectrodes were inserted in the hippocampus CA1 region of 21 Institute of Cancer Research mice. The mice were divided into 3 groups, including an EtOH injection group (1.5 g/kg), a saline injection group (1.5 g/kg), and a negative control group that received no injection. A data acquisition system was employed to detect the local field potentials (LFPs) and spike potentials following acute EtOH administration. Various multichannel electrophysiological signals were collected in vivo in each group at 60 minutes, from which the firing rate and wavelet entropy (WE) were analyzed further. RESULTS: Firing rates began to decline at 20 minutes postinjection and then gradually recovered from 40 to 60 minutes. In contrast, 20 minutes post-injection, WE increased maximally and then returned to normal from 40 to 60 minutes (p < 0.05). Pronounced changes in the relative energy of theta and alpha oscillations were also observed after 20 minutes of alcohol exposure and recovery occurred by 60 minutes (p < 0.05). CONCLUSIONS: A major mechanism of EtOH's action on the hippocampus is neurotransmitter blocking in the form of excitatory neuron inhibition in vivo. Changes in hippocampal spikes coincided with changes in LFPs during the entire time course of acute EtOH administration. The correlation between spikes and LFPs suggests that they jointly affect encoding in hippocampus.

Assays on 3D tumor spheroids for exploring the light dosimetry of photodynamic effects under different gaseous conditions.

The multifaceted nature of photodynamic therapy (PDT) requires a throughout evaluation of a multitude of parameters when devising preclinical protocols. In this study, we constructed MCF-7 human breast tumor spheroid assays to infer PDT irradiation doses at four gradient levels for violet light at 408 nm and red light at 625 nm under normal and hypoxic oxygen conditions. The compacted three-dimensional (3D) tumor models conferred PDT resistance as compared to monolayer cultures due to heterogenous distribution of photosensitizers along with the presence of internal hypoxic region. Cell viability results indicated that the violet light was more efficient to kill cells in the spheroids under normal oxygen conditions, while cells exposed to the hypoxic microenvironment exhibited minimal PDT-induced death. The combination of 3D tumor spheroid assays and the multiparametric screening platform presented a solid framework for assessing PDT efficacy across a wide range of different physiological conditions and therapeutic regimes.

Optical-resolution photoacoustic microelastography system for elasticity mapping: Phantom study and practical application.

Elastography is a noninvasive technique for characterizing the mechanical properties of biological tissues. Conventional methods have limitations in resolution and sensitivity, hindering disease detection in clinical diagnostics. To address these issues, this study developed an optical-resolution photoacoustic microelastography (OR-PAME) system. Using an agar tissue phantom with varying agar concentrations and contrast agents, PAME evaluated elasticity distribution under compression in both lateral and axial dimensions. It indirectly measured elastic properties by correlating photoacoustic responses, temporal lags, and induced displacement. We also applied the system to the study of the distribution of elastic characteristics of the liver tissue after ablation, which confirmed the potential of OR-PAME in the study of elastic characteristics. Quantitative analysis showed greater lateral displacement in regions with reduced agar concentrations, indicating decreased stiffness. PAME also detected vertical displacement along the axial plane, validating its efficacy in elastographic imaging. By improving resolution and penetration, PAME provides superior visualization of elasticity distribution. Its methodology correlates microstructural alterations with tissue biomechanics, holding potential implications in medical diagnostics.

Multi-channel Small Animal Drug Metabolism Real-Time Monitoring Fluorescence System.

PURPOSE: The data acquisition of drug metabolism analysis requires a lot of time and animal resources. However, there are often many deviations in the results of pharmacokinetic analysis. Conventional methods cannot measure the blood drug concentration data in multiple tissues at the same time, and the data is obtained by in vitro measurement, which produces time errors, in vitro data errors, and individual differences between animals. In the analysis of pharmacokinetic parameters, it will seriously affect the pass rate of clinical trials of R&D drugs and the accuracy of the dosing schedule. To the best of our knowledge, we have not found the study of in vivo blood drug concentration using multi-channel equipment. Therefore, the purpose of this paper is to build a set of multi-organ monitoring and analysis instruments for synchronously monitoring the metabolism of drugs in various tissues of small animals, so as to obtain real in vivo data of blood drug concentration in real time. PROCEDURES: Using the fluorescence properties and laser-induced fluorescence principle of drugs, we designed six channels to monitor the changes of fluorescence-labeled drugs in their main metabolic organs, a multi-channel calibration method was proposed to improve the accuracy of the time-division multiplexing, the real-time collection of drug concentration in vivo is realized, and the drug metabolism curve in vivo can be observed. RESULTS: The instrument satisfies the collection of small doses of drugs such as microgram; the detection sensitivity can reach 10 ng/ml, and can monitor and collect the drug metabolism of multiple small animal tissues at the same time, which greatly reduces the use of animals, reduces the differences between individuals, and reduces consumption cost and improve the detection efficiency of parameters, and obtain data information that is closer to the real biology. CONCLUSION: The real-time continuous monitoring and data collection of the drug metabolism in the plasma of living small animals and the important organs such as kidney, liver, and spleen were realized. The research and development of new drugs and clinical research have higher practical value.

Fiber optic-based integrated system for in vivo multiscale pharmacokinetic monitoring.

This paper presents the development of a fiber-optic-based fluorescence detection system for multi-scale monitoring of drug distribution in living animals. The integrated system utilized dual laser sources at the wavelengths of 488 nm and 650 nm and three photomultiplier channels for multi-color fluorescence detection. The emission spectra of fluorescent substances were tracked using the time-resolved fluorescence spectroscopy module to continuously monitor their blood kinetics. The fiber bundle, consisting of 30,000 optic filaments, was designed for wide-field mesoscopic imaging of the drug's interactions within organs. The inclusion of a gradient refractive index (GRIN) lens within the setup enabled fluorescence confocal laser scanning microscopy to visualize the drug distribution at the cellular level. The system performance was verified by imaging hepatic and renal tissues in mice using cadmium telluride quantum dots (CdTe QDs) and R3. By acquiring multi-level images and real-time data, our integrated system underscores its potential as a potent tool for drug assessment, specifically within the realms of pharmacokinetic and pharmacodynamic investigations.

Molecular detection and characterization of Coxiella burnetii in aborted samples of livestock in China.

Coxiella burnetii is the causative agent of zoonotic Q fever. Animals are the natural reservoirs of C. burnetii, and domestic livestock represent the major sources of human infection. C. burnetii infection in pregnant females may causes abortion during late pregnancy, whereby massive shedding of C. burnetii with abortion products becomes aerosolized and persists in the environment. Therefore, monitoring and surveillance of this infection in livestock is important for the prevention of the C. burnetii transmission. Previous serological surveys have shown that C. burnetii infection is endemic in livestock in China. However, few data are available on the diagnosis of C. burnetii as a cause of abortion by molecular methods in livestock. To get a better understanding of the impact of C. burnetii infection on domestic livestock in China, a real-time PCR investigation was carried out on collected samples from different domestic livestock suffering abortion during 2021-2023. A total of 338 samples collected from eight herds of five livestock species were elected. The results showed that 223 (66 %) of the collected samples were positive for C. burnetii DNA using real-time PCR. For the aborted samples, 82 % (128/15) of sheep, 81 % (34/42) of goats, 44 % (15/34) of cattle, 69 % (18/26) of camels, and 50 % (17/34) of donkeys were positive for C. burnetii. Besides, 44 % (8/18) and 4 % (1/25) of asymptomatic individuals of sheep and donkey were also positive for C. burnetii. In addition, the positive samples were further confirmed by amplification and sequencing of the C. burnetii-specific isocitrate dehydrogenase (icd) gene. Phylogenetic analysis based on specific gene fragments of icd genes revealed that the obtained sequences in this study were clustered into two different groups associated with different origin of hosts and geographic regions. This is the first report confirming that C. burnetii exists in aborted samples of sheep, goats, cattle, donkeys and camels in China. Further studies are needed to fully elucidate the epidemiology of this pathogen in livestock as well as the potential risks to public health.

Exploring the neural correlates of fat taste perception and discrimination: Insights from electroencephalogram analysis.

Understanding neural pathways and cognitive processes involved in the transformation of dietary fats into sensory experiences has profound implications for nutritional well-being. This study presents an efficient approach to comprehending the neural perception of fat taste using electroencephalogram (EEG). Through the examination of neural responses to different types of fatty acids (FAs) in 45 participants, we discerned distinct neural activation patterns associated with saturated versus unsaturated fatty acids. The spectrum analysis of averaged EEG signals revealed notable variations in δ and α-frequency bands across FA types. The topographical distribution and source localization results suggested that the brain encodes fat taste with specific activation timings in primary and secondary gustatory cortices. Saturated FAs elicited higher activation in cortical associated with emotion and reward processing. This electrophysiological evidence enhances our understanding of fundamental mechanisms behind fat perception, which is helpful for guiding strategies to manage hedonic eating and promote balanced fat consumption.

All-in-one device for mapping the interactive effects of photodynamic therapy dosimetry in tumor gaseous microenvironment.

Photodynamic therapy (PDT) elicits cell death, vascular damage, or/and anti-tumor host immune response upon activating the administered photosensitive drug by an appropriate light source. Because PDT is heavily dependent on tissue oxygen (O2) in essence, the concentration-dependent impact of O2 on tailoring cellular response to PDT remains an in-depth investigation. As a multifaceted modality, optimal combinations of photosensitizer (PS) concentration, light dose, and O2 delivery are critical to achieve ideal therapeutic outcomes. We herein present a fully integrated all-in-one device for the in vitro assessment of PDT efficacy synchronizing the quantitative control of three PDT disciplines simultaneously, aiming at 1) identifying the influence of varying gaseous microenvironments on PDT; and 2) determining the contribution of each PDT factor and estimating the strength of their synergic effect. The gas-gradient-generating unit for contactless headspace O2 delivery and spatial light control filtering layer in our device could either work as a stand-alone module or combine to screen a range of experimental PDT parameters. By sweeping a total of 128 conditions over four 5-aminolevulinic acid (5-ALA) concentrations, four light dosages, and eight O2 levels in one single experiment, we determined the main effects of the three key PDT agents and highlighted the interactive effect between 5-ALA and light after full-factorial statistical analysis. Our device is not only a versatile tool for predicting PDT efficacy during the translational study but also provides valuable multidimensional information for the interrelation between key PDT factors, which may expedite clinical PDT dosimetry and furnish new insights for the fundamental understanding of photobiological processes.

Quantitative proteomics analysis on the meat quality of processed pale, soft, and exudative (PSE)-like broiler pectoralis major by different heating methods.

This study aims to assess and compare the influences of different heating methods on the quality characteristics of pale, soft, and exudative (PSE)-like and normal (NOR) pectoralis major through quantitative proteomic analysis. A total of 632 proteins were identified, and there were 84, 89, 50, and 43 differentially abundant proteins (DAPs) between processed PSE and NOR samples after four thermal treatments, including boiling (BO), steaming (ST), roasting (RO), and microwaving (MV), respectively, where moist heating conditions led to more different protein abundance. Processed PSE muscles resulted in significant changes in structural proteins related to myofibrillar and connective tissue, which could be associated with their structural instability and degraded quality. Collagen, tropomyosin, myoglobin, and hemoglobin could be potential indicators of PSE muscles color stability and variation during thermal processing. The quantitative proteomic analysis will help correlate molecular changes with processed meat quality towards future optimization of PSE poultry meat processing.

Insights into brain perceptions of the different taste qualities and hedonic valence of food via scalp electroencephalogram.

Investigating brain activity is essential for exploring taste-experience related cues. The paper aimed to explore implicit (unconscious) emotional or physiological responses related to taste experiences using scalp electroencephalogram (EEG). We performed implicit measures of tastants of differing perceptual types (bitter, salty, sour and sweet) and intensities (low, medium, and high). The results showed that subjects were partially sensitive to different sensory intensities, i.e., for high intensities, taste stimuli could induce activation of different rhythm signals in the brain, with α and θ bands possibly being more sensitive to different taste types. Furthermore, the neural representations and corresponding sensory qualities (e.g., "sweet: pleasant" or "bitter: unpleasant") of different tastes could be discriminated at 250-1,500 ms after stimulus onset, and different tastes exhibited distinct temporal dynamic differences. Source localization indicated that different taste types activate brain areas associated with emotional eating, reward processing, and motivated tendencies, etc. Overall, our findings reveal a larger sophisticated taste map that accounted for the diversity of taste types in the human brain and assesses the emotion, reward, and motivated behavior represented by different tastes. This study provided basic insights and a perceptual foundation for the relationship between taste experience-related decisions and the prediction of brain activity.

Size-dependent neurotoxicity of micro- and nanoplastics in flowing condition based on an in vitro microfluidic study.

With the widespread presence of plastic wastes, knowledge about the potential environmental risks and bioavailability of micro- or nanoplastics fragmented from large analogs is of utmost importance. As the particle size matters in mediating endocytic mechanism and particle internalization, we first studied the effects of polystyrene microparticles (PS-MPs, 1 µm) and polystyrene nanoparticles (PS-NPs, 100 nm) of two different sizes at varying concentrations of 5, 25 and 75 µg/mL on the mouse hippocampal neuronal HT22 cells. The in vitro study showed efficient cellular uptake of PS-MPs and PS-NPs of both sizes. The adverse effects of cellular metabolic activity as reflective of excess Reactive Oxygen Species (ROS) and cell cycle S phase arresting were observed especially at the greater concentration of smaller-sized PS particles, consequently leading to mild cytotoxicity. We further evaluated the dynamic particle-cell interaction with a continuous supply of PS particles using a microfluidic device. By recapitulating the in vivo mechanical microenvironments while allowing homogeneous distribution of PS particles, the dynamic exposure to PS particles of both sizes under flowing conditions resulted in much lesser viability of neural cells than the traditional static exposure. As the flowing dynamics may avoid the gravitational settling of particles and allow more efficient cellular uptake, the size distribution, together with the exposure configurations, contributed significantly to the determination of the PS particle cytotoxicity. The on-chip investigation and a better understanding of particle translocation mechanisms would offer very much to the risk assessment of PS particles on human health.

Impacts of Catch-Up Immunization program with the 13-Valent pneumococcal Conjugate vaccine in Taiwan: Focus on age-stratified differences and high-risk population (2001-2015).

BACKGROUND: Taiwan commenced a national catch-up immunization program with a 13-valent pneumococcal conjugate vaccine (PCV13) in 2013 for children aged 2-5 years old and in 2014 for children aged 1-5 years old. However, real-world nationwide evidence of both the direct protection and indirect protection of all-cause pneumonia and pneumococcal pneumonia has been scarce, especially among high-risk populations, defined as patients with chronic diseases or immunosuppression. The aim of this study was to examine the impact of the national PCV13 catch-up program on all-cause pneumonia and pneumococcal pneumonia among overall and high-risk populations using interrupted time series analysis. METHODS: Using the National Health Insurance Research Database (NHIRD) from January 2001 to December 2015, we assessed the impact of this catch-up program by interrupted time-series analyses age-stratified (0-1, 2-4, 5-9, 10-17, 18-34, 35-49, 50-64, 65 + years old) incidence of pneumococcal pneumonia and all-cause pneumonia (100,000 person-quarter) among the overall and high-risk populations. RESULTS: The impact of this program was most profound on the incidence of pneumococcal pneumonia in children aged 2-4 years old (level change -10.56 per 100,000 person-quarters, p = 0.04; trend change -2.93, p less than 0.01). Indirect protection among unvaccinated children (0-1 years old: trend change -1.19, p = 0.01; 5-9 years old: trend change -1.04, p = 0.03; 10-17 years old: level change -1.42 per 100,000 person-quarters, p = 0.03) was also found. The incidence of all-cause pneumonia also decreased in children aged 2-4 (level change -234.91 per 100,000 person-quarter, p = 0.058) and 5-9 years old (level change -173.96 per 100,000 person-quarter, p = 0.0424). However, we did not find a significant impact among most high-risk populations. CONCLUSIONS: Our study suggests that the introduction of this catch-up program with PCV13 was associated with significant declines in the incidence of pneumococcal pneumonia and all-cause pneumonia in vaccinated children, and indirect protection from the program was also found in unvaccinated children.