Acoustic Cell Patterning for Structured Cell-Laden Hydrogel Fibers/Tubules.

Cell-laden hydrogel fibers/tubules are one of the fundamentals of tissue engineering. They have been proven as a promising method for constructing biomimetic tissues, such as muscle fibers, nerve conduits, tendon and vessels, etc. However, current hydrogel fiber/tubule production methods have limitations in ordered cell arrangements, thus impeding the biomimetic configurations. Acoustic cell patterning is a cell manipulation method that has good biocompatibility, wide tunability, and is contact-free. However, there are few studies on acoustic cell patterning for fiber production, especially on the radial figure cell arrangements, which mimic many native tissue-like cell arrangements. Here, an acoustic cell patterning system that can be used to produce hydrogel fibers/tubules with tunable cell patterns is shown. Cells can be pre-patterned in the liquid hydrogel before being extruded as cross-linked hydrogel fibers/tubules. The radial patterns can be tuned with different complexities based on the acoustic resonances. Cell viability assays after 72 h confirm good cell viability and proliferation. Considering the biocompatibility and reliability, the present method can be further used for a variety of biomimetic fabrications.

Micro/nanosystems for controllable drug delivery to the brain.

Drug delivery to the brain is crucial in the treatment for central nervous system disorders. While significant progress has been made in recent years, there are still major challenges in achieving controllable drug delivery to the brain. Unmet clinical needs arise from various factors, including controlled drug transport, handling large drug doses, methods for crossing biological barriers, the use of imaging guidance, and effective models for analyzing drug delivery. Recent advances in micro/nanosystems have shown promise in addressing some of these challenges. These include the utilization of microfluidic platforms to test and validate the drug delivery process in a controlled and biomimetic setting, the development of novel micro/nanocarriers for large drug loads across the blood-brain barrier, and the implementation of micro-intervention systems for delivering drugs through intraparenchymal or peripheral routes. In this article, we present a review of the latest developments in micro/nanosystems for controllable drug delivery to the brain. We also delve into the relevant diseases, biological barriers, and conventional methods. In addition, we discuss future prospects and the development of emerging robotic micro/nanosystems equipped with directed transportation, real-time image guidance, and closed-loop control.

Golden section criterion to achieve droplet trampoline effect on metal-based superhydrophobic surface.

Clarifying the consecutive droplet rebound mechanisms can provide scientific inspirations to regulate dynamic wettability of superhydrophobic surface, which facilitates the practical applications on efficient heat control and active anti-icing. Generally, droplet rebound behaviors are directly affected by surface structure and Weber number. Here, we report a novel "golden section" design criterion to regulate the droplet rebound number determined by the structure spacing, subverting conventional knowledge. Especially, the droplet can continuously rebound for 17 times on the metal-based surface, exhibiting an amazing phenomenon of "droplet trampoline". The droplet rebound number has been experimentally revealed to be closely related to Weber number. We propose novel quantitative formulas to predict droplet rebound number and clarify the coupling effect of the structure spacing and the Weber number on the rebound mechanisms, which can be utilized to establish the regulation criteria of rebound numbers and develop novel metal-based superhydrophobic materials.

Applications of flexible electronics related to cardiocerebral vascular system.

Ensuring accessible and high-quality healthcare worldwide requires field-deployable and affordable clinical diagnostic tools with high performance. In recent years, flexible electronics with wearable and implantable capabilities have garnered significant attention from researchers, which functioned as vital clinical diagnostic-assisted tools by real-time signal transmission from interested targets in vivo. As the most crucial and complex system of human body, cardiocerebral vascular system together with heart-brain network attracts researchers inputting profuse and indefatigable efforts on proper flexible electronics design and materials selection, trying to overcome the impassable gulf between vivid organisms and rigid inorganic units. This article reviews recent breakthroughs in flexible electronics specifically applied to cardiocerebral vascular system and heart-brain network. Relevant sensor types and working principles, electronics materials selection and treatment methods are expounded. Applications of flexible electronics related to these interested organs and systems are specially highlighted. Through precedent great working studies, we conclude their merits and point out some limitations in this emerging field, thus will help to pave the way for revolutionary flexible electronics and diagnosis assisted tools development.

16S full-length gene sequencing analysis of intestinal flora in breast cancer patients in Hainan Province.

Breast cancer has become the number one cancer in the world, and intestinal flora may be closely linked to it. Geographic location also has an important impact on human intestinal flora. We conducted the first study on the intestinal flora of breast cancer patients and non-breast cancer patients in a tropical region - Hainan Province in China. At the same time, Pacbio platform based on third-generation sequencing was used for the first time to conduct 16S full-length sequencing of fecal microorganism DNA. We completed the species diversity analysis and differential species analysis of the intestinal flora between the two groups, inferred their functional genetic composition and performed functional difference analysis. There were statistically significant differences in alpha diversity between the two groups in Hainan Province. By species composition difference analysis, at the phylum level, Bacteroidales (P = 0.006) and Firmicutes (P = 0.002) was differed between the two groups, and at the genus level, 17 breast cancer-related differential species such as Bacteroides were screened. According to the five grouping methods including ER level, PR level, HER2 status, Ki67 index and histological grade of breast cancer patients, 4, 1, 9, 6, 5 differential microbiota were screened out respectively, which were in total 25 (P < 0.05 for all subgroups) . The functional prediction and difference analysis revealed two functional metabolisms with significant differences between the two groups of microbes (P < 0.05). These results suggest that breast cancer is associated with changes in the composition and function of intestinal flora. These microflora and functional differences may become biomarkers or new targets for diagnosis and treatment of breast cancer.

Coupled Effect of Low Temperature and Electrolyte Immersion on the Tensile Properties of Separators in Lithium-Ion Batteries.

The performance degradation at low temperatures and frequent safety accidents have aggravated security risks and inhibited the long-term service of lithium-ion batteries (LIBs). As a key component of LIBs, the separator has a great impact on the performance and safety of the battery. In this study, tensile tests of two commercial polyolefin separators (Celgard 2325 and Celgard PE) are performed under low-temperature and immersion conditions. Four representative temperature points and dimethyl carbonate [(DMC), the common solvent in electrolytes] are selected to investigate the coupling effect on the mechanical properties of the separators. The results show that both the separators have anisotropy, but the performance of Celgard 2325 varies more significantly than that of Celgard PE along different directions. Additionally, it is found that with a decrease in the temperature, the tensile strength of the two separators increases, while the elongation decreases. Electrolyte immersion induces a softening tendency in Celgard 2325. Due to the special effect of the residual electrolyte on polyethylene fibers, Celgard PE shows the opposite result. Furthermore, the effect of low temperature is revealed by the analysis of the crystallinity and molecular structure, which can be obtained by X-ray diffraction and Raman spectroscopy, respectively. In addition, the contact angle is measured to describe the wettability variation related to low temperature. The present work provides a theoretical basis and experimental data for the application and development of separators.

Low Expression of BIRC5-206 Promotes Cancer Progression in Nasopharyngeal Carcinoma via Enhancing Expression of Stem Cell Markers.

OBJECTIVE: Survivin is highly expressed in various malignant tumor cells and positively related to poor prognosis and drug resistance. This study aimed to explore the role of non-coding splice variant of Survivin, BIRC5-206 (ENST00000589892.1) in the progression of nasopharyngeal carcinoma (NPC), a malignant tumor that highly occurs in the southern region of China. METHODS: shRNA was used to knockdown BIRC5-206 mRNA level in CNE-2 and HOPNE-1 cells. Then, cell death, migration, invasion and clone formation ability of CNE-2 and HOPNE-1 cells were detected by flow cytometry, scratch-healing experiments, transwell invasion assay and clone formation assay, respectively. CD44+ and CD133+ positive cells were determined via Flow cytometry. Oct4, Nanog and SOX2 protein levels in CNE-2 and HOPNE-1 cells were measured by Western blot. RESULTS: BIRC5-206 decreased significantly in NPC cell lines. Silencing of BIRC5-206 suppressed the apoptosis, facilitated the migration, invasion and proliferation of both HONE1 and CNE-2 cells. In addition, knockdown of BIRC5-206 significantly promoted the expression cancer stem cell marker (CD44 and CD133) and pluripotency markers (Oct4, Sox2 and Nanog). CONCLUSIONS: BIRC5-206 might facilitate NPC tumor progression by inducing the transformation of NPC cells to cancer stem cells.

Improving the Luminescence and Stability of Carbon-Centered Radicals by Kinetic Isotope Effect.

The kinetic isotope effect (KIE) is beneficial to improve the performance of luminescent molecules and relevant light-emitting diodes. In this work, the influences of deuteration on the photophysical property and stability of luminescent radicals are investigated for the first time. Four deuterated radicals based on biphenylmethyl, triphenylmethyl, and deuterated carbazole were synthesized and sufficiently characterized. The deuterated radicals exhibited excellent redox stability, as well as improved thermal and photostability. The appropriate deuteration of relevant C-H bonds would effectively suppress the non-radiative process, resulting in the increase in photoluminescence quantum efficiency (PLQE). This research has demonstrated that the introduction of deuterium atoms could be an effective pathway to develop high-performance luminescent radicals.

Riboflavin protects against heart failure via SCAD-dependent DJ-1-Keap1-Nrf2 signalling pathway.

BACKGROUND AND PURPOSE: Our recent studies have shown that flavin adenine dinucleotide (FAD) exerts cardiovascular protective effects by supplementing short-chain acyl-CoA dehydrogenase (SCAD). The current study aimed to elucidate whether riboflavin (the precursor of FAD) could improve heart failure via activating SCAD and the DJ-1-Keap1-Nrf2 signalling pathway. EXPERIMENTAL APPROACH: Riboflavin treatment was given to the mouse transverse aortic constriction (TAC)-induced heart failure model. Cardiac structure and function, energy metabolism and apoptosis index were assessed, and relevant signalling proteins were analysed. The mechanisms underlying the cardioprotection by riboflavin were analysed in the cell apoptosis model induced by tert-butyl hydroperoxide (tBHP). KEY RESULTS: In vivo, riboflavin ameliorated myocardial fibrosis and energy metabolism, improved cardiac dysfunction and inhibited oxidative stress and cardiomyocyte apoptosis in TAC-induced heart failure. In vitro, riboflavin ameliorated cell apoptosis in H9C2 cardiomyocytes by decreasing reactive oxygen species (ROS). At the molecular level, riboflavin significantly restored FAD content, SCAD expression and enzymatic activity, activated DJ-1 and inhibited the Keap1-Nrf2/HO1 signalling pathway in vivo and in vitro. SCAD knockdown exaggerated the tBHP-induced DJ-1 decrease and Keap1-Nrf2/HO1 signalling pathway activation in H9C2 cardiomyocytes. The knockdown of SCAD abolished the anti-apoptotic effects of riboflavin on H9C2 cardiomyocytes. DJ-1 knockdown hindered SCAD overexpression anti-apoptotic effects and regulation on Keap1-Nrf2/HO1 signalling pathway in H9C2 cardiomyocytes. CONCLUSIONS AND IMPLICATIONS: Riboflavin exerts cardioprotective effects on heart failure by improving oxidative stress and cardiomyocyte apoptosis via FAD to stimulate SCAD and then activates the DJ-1-Keap1-Nrf2 signalling pathway.

The Ovs surface defecting of an S-scheme g-C3N4/H2Ti3O7 nanoheterostructures with accelerated spatial charge transfer.

Semiconductor photocatalysis was a rising star in the sustainable transformation of solar energy for environmental problems governance. Herein, an S-scheme g-C3N4/H2Ti3O7 heterostructure was constructed and applied to tetracycline hydrochloride (TCH) destruction. The g-C3N4/H2Ti3O7 composite has a superior photocatalytic property to degrade TCH in contrast with bare g-C3N4 and H2Ti3O7. The 20% g-C3N4/H2Ti3O7 (CNHTO20) composite exhibited the optimum photocatalytic performance, and the degradation efficiency of 20 mg/L TCH reached 87.37% within 3 h (K = 0.572 min-1). The affluent active sites of the g-C3N4 nanosheet and effective interfacial charge separation of the S-scheme pathway facilitated the excellent performance. Moreover, the ample oxygen vacancies (Ovs) act as the electron mediator, not only reducing the band gap energy by producing the formation of defect levels, but also broadening the photo response range and promoting the interfacial charge transfer. The coordination complexes formed between TCH molecules and Ti (IV) ions in CNHTO20 composites induce strong visible light absorption through ligand-metal charge transfer (LMCT). The Ti4+/Ti3+ metal cycle in CNHTO20 was conducive to the separation of the photogenerated electron-hole pairs on the heterojunction interface as well. The ESR characterization and trapping experiments certified that the dominant substances were OH, O2- and h+. The AQY calculated by the COD removal rate was 0.16%. Conclusively, the S-scheme heterojunction between H2Ti3O7 and g-C3N4 enabled the CNHTO photocatalyst with high redox ability and boosted photocatalytic performance accordingly. This study may shed some enlightenment on the construction of heterojunctions and the realistic treatment of wastewater.

Manipulation with sound and vibration: A review on the micromanipulation system based on sub-MHz acoustic waves.

Manipulation of micro-objects have been playing an essential role in biochemical analysis or clinical diagnostics. Among the diverse technologies for micromanipulation, acoustic methods show the advantages of good biocompatibility, wide tunability, a label-free and contactless manner. Thus, acoustic micromanipulations have been widely exploited in micro-analysis systems. In this article, we reviewed the acoustic micromanipulation systems that were actuated by sub-MHz acoustic waves. In contrast to the high-frequency range, the acoustic microsystems operating at sub-MHz acoustic frequency are more accessible, whose acoustic sources are at low cost and even available from daily acoustic devices (e.g. buzzers, speakers, piezoelectric plates). The broad availability, with the addition of the advantages of acoustic micromanipulation, make sub-MHz microsystems promising for a variety of biomedical applications. Here, we review recent progresses in sub-MHz acoustic micromanipulation technologies, focusing on their applications in biomedical fields. These technologies are based on the basic acoustic phenomenon, such as cavitation, acoustic radiation force, and acoustic streaming. And categorized by their applications, we introduce these systems for mixing, pumping and droplet generation, separation and enrichment, patterning, rotation, propulsion and actuation. The diverse applications of these systems hold great promise for a wide range of enhancements in biomedicines and attract increasing interest for further investigation.

Full Regional Creep Displacement Map of Articular Cartilage Based on Nanoindentation Array.

The elucidation of the mechanisms underlying articular cartilage lesions poses a formidable challenge in the field of cartilage repair. Despite significant strides in cartilage mechanics research, the region-dependent creep properties of articular cartilage remain elusive. In this study, we employ depth-sensing indentation tests to experimentally determine the creep properties of four distinct regions of articular cartilage, thereby unveiling a region-dependent full map of creep parameters. The measured creep displacement-time response curves indicate that the creep properties of the articular cartilage exhibit a clear regional correlation. Accordingly, the full regional creep map of articular cartilage is experimentally constructed for the first time. The correlation between the microstructures and the creep properties of cartilage in different regions is revealed. A three-parameter model is established to describe the creep velocity-displacement response of cartilage. Raman spectra reveal that the proteoglycan content is positively correlated with creep properties. The Raman shift directly indicates diverse residual stresses in different microregions. The obtained data facilitate a deep understanding of the potential creep dependent damage mechanism of cartilage and the further development of artificial cartilage materials.

Adenosine 2A receptor contributes to the facilitation of post-infectious irritable bowel syndrome by γδ T cells via the PKA/CREB/NF-κB signaling pathway.

BACKGROUND: Immunological dysfunction-induced low-grade inflammation is regarded as one of the predominant pathogenetic mechanisms in post-infectious irritable bowel syndrome (PI-IBS). γδ T cells play a crucial role in innate and adaptive immunity. Adenosine receptors expressed on the surface of γδ T cells participate in intestinal inflammation and immunity regulation. AIM: To investigate the role of γδ T cell regulated by adenosine 2A receptor (A2AR) in PI-IBS. METHODS: The PI-IBS mouse model has been established with Trichinella spiralis (T. spiralis) infection. The intestinal A2AR and A2AR in γδ T cells were detected by immunohistochemistry, and the inflammatory cytokines were measured by western blot. The role of A2AR on the isolated γδ T cells, including proliferation, apoptosis, and cytokine production, were evaluated in vitro. Their A2AR expression was measured by western blot and reverse transcription polymerase chain reaction (RT-PCR). The animals were administered with A2AR agonist, or A2AR antagonist. Besides, γδ T cells were also injected back into the animals, and the parameters described above were examined, as well as the clinical features. Furthermore, the A2AR-associated signaling pathway molecules were assessed by western blot and RT-PCR. RESULTS: PI-IBS mice exhibited elevated ATP content and A2AR expression (P < 0.05), and suppression of A2AR enhanced PI-IBS clinical characteristics, indicated by the abdominal withdrawal reflex and colon transportation test. PI-IBS was associated with an increase in intestinal T cells, and cytokine levels of interleukin-1 (IL-1), IL-6, IL-17A, and interferon-α (IFN-α). Also, γδ T cells expressed A2AR in vitro and generated IL-1, IL-6, IL-17A, and IFN-α, which can be controlled by A2AR agonist and antagonist. Mechanistic studies demonstrated that the A2AR antagonist improved the function of γδ T cells through the PKA/CREB/NF-κB signaling pathway. CONCLUSION: Our results revealed that A2AR contributes to the facilitation of PI-IBS by regulating the function of γδ T cells via the PKA/CREB/NF-κB signaling pathway.

Research on emergency material demand based on urgency and satisfaction under public health emergencies.

In recent years, the frequent occurrence of public health emergencies has had a significant impact on people's life. The study of emergency logistics has also attracted scholars' attention. Therefore, matching emergency materials' supply and demand quickly, which meets urgency and satisfaction, is the purpose of this paper. This paper used the Metabolism Grey Model (1,1) (GM (1,1)) and the material demand prediction model to predict the number of infections and material demand. Besides, we established a bi-objective optimization model by constructing a profit and loss matrix and a comprehensive utility perception matrix. The results show that the method is helpful in matching the optimal supply and demand decision quickly on the basis of meeting urgency and satisfaction. The method is helpful in improving the fairness of emergency material distribution, which could better protect people's livelihoods.

Short-chain acyl-CoA dehydrogenase is a potential target for the treatment of vascular remodelling.

OBJECTIVES: Short-chain acyl-CoA dehydrogenase (SCAD), a key enzyme in the fatty acid oxidation process, is not only involved in ATP synthesis but also regulates the production of mitochondrial reactive oxygen species (ROS) and nitric oxide synthesis. The purpose of this study was to investigate the possible role of SCAD in hypertension-associated vascular remodelling. METHODS: In-vivo experiments were performed on spontaneously hypertensive rats (SHRs, ages of 4 weeks to 20 months) and SCAD knockout mice. The aorta sections of hypertensive patients were used for measurement of SCAD expression. In-vitro experiments with t-butylhydroperoxide (tBHP), SCAD siRNA, adenovirus-SCAD (MOI 90) or shear stress (4, 15 dynes/cm 2 ) were performed using human umbilical vein endothelial cells (HUVECs). RESULTS: Compared with age-matched Wistar rats, aortic SCAD expression decreased gradually in SHRs with age. In addition, aerobic exercise training for 8 weeks could significantly increase SCAD expression and enzyme activity in the aortas of SHRs while decreasing vascular remodelling in SHRs. SCAD knockout mice also exhibited aggravated vascular remodelling and cardiovascular dysfunction. Likewise, SCAD expression was also decreased in tBHP-induced endothelial cell apoptosis models and the aortas of hypertensive patients. SCAD siRNA caused HUVEC apoptosis in vitro , whereas adenovirus-mediated SCAD overexpression (Ad-SCAD) protected against HUVEC apoptosis. Furthermore, SCAD expression was decreased in HUVECs exposed to low shear stress (4 dynes/cm 2 ) and increased in HUVECs exposed to 15 dynes/cm 2 compared with those under static conditions. CONCLUSION: SCAD is a negative regulator of vascular remodelling and may represent a novel therapeutic target for vascular remodelling.

Construction of an S-scheme TiOF2/HTiOF3 heterostructures with abundant OVs and OH groups: Performance, kinetics and mechanism insight.

Developing efficient photocatalysts is of crucial significance for the development of photocatalysis techniques. In this work, an S-scheme alkaline-washed TiOF2/HTiOF3(OHTOF) heterostructures with abundant Oxygen vacancies (Ovs) and OH groups was successfully constructed and used to remedy antibiotic wastewater under simulated sunlight. The generation of HTiOF3 was induced by g-C3N4 regulation. The results displayed that OHTOF15 composite possessed the best photocatalytic performance, which could degrade 94.2% tetracyclinehydrochloride (TCH) at a rate speed constant of 1.077 min-1 in 2.5 h. The after-alkali-washing process increased the concentration of OH groups and Ovs defects, and greatly enlarged the surface area. The abundant Ovs and OH groups were conducive to the formation of free radicals' and the transport of charge carriers. Compared with the pristine TiOF2, the absorption sidebands of OHTOF series were greatly red-shifted, which indicated that the increase of OH groups and the etching of the morphology of OHTOF further enhanced its visible-light harvesting ability. Furthermore, the metal cycle of the variable state of Ti4+/Ti3+ in OHTOF15 compensated for the charge balance and promoted the efficient separation of the carriers. Additionally, the apparent quantum efficiency (AQE) of the TCH photodegradation system based on Chemical Oxygen Demand (COD) removal efficiency was calculated to be 0.32%. It was confirmed that the electron transport path in TiOF2/HTiOF3 nanocomposites system followed the S-scheme type, which increased the charge carriers' separation rate and maintained a strong redox capacity. This work could provide some enlightenment for the construction of the semiconducting heterojunction and controllable surface defects engineering.

Biomolecular actuators for genetically selective acoustic manipulation of cells.

The ability to physically manipulate specific cells is critical for the fields of biomedicine, synthetic biology, and living materials. Ultrasound has the ability to manipulate cells with high spatiotemporal precision via acoustic radiation force (ARF). However, because most cells have similar acoustic properties, this capability is disconnected from cellular genetic programs. Here, we show that gas vesicles (GVs)-a unique class of gas-filled protein nanostructures-can serve as genetically encodable actuators for selective acoustic manipulation. Because of their lower density and higher compressibility relative to water, GVs experience strong ARF with opposite polarity to most other materials. When expressed inside cells, GVs invert the cells' acoustic contrast and amplify the magnitude of their ARF, allowing the cells to be selectively manipulated with sound waves based on their genotype. GVs provide a direct link between gene expression and acoustomechanical actuation, opening a paradigm for selective cellular control in a broad range of contexts.

Sound-Speed Modifying Acoustic Metasurfaces for Acoustic Holography.

Acoustic metasurfaces offer unique capabilities to steer and direct acoustic fields, though these are generally composed of complex 3D structures, complicating their fabrication and applicability to higher frequencies. Here, an ultrathin metasurface approach is demonstrated, wherein planarized micropillars in a discretized phase array are utilized. This subwavelength metasurface is easily produced via a single-step etching process and is suitable for megahertz-scale applications. The flexibility of this approach is further demonstrated in the production of complex acoustic patterns via acoustic holography. This metasurface approach, with models used to predict their behavior, has broad potential in applications where robust, high-frequency acoustic manipulation is required, including microfluidics, cell/tissue engineering, and medical ultrasound.

Association of Alcohol Drinking and Helicobacter pylori Infection : A Meta-analysis.

BACKGROUND: The association between drinking and Helicobacter pylori infection was not clear in the literature. Owing to mixed and inconclusive results, a meta-analysis was conducted to summarize and clarify this association systematically. METHODS: Based on a comprehensive search of PubMed, Embase, and Web of Science databases, studies investigating the association between drinking and H. pylori infection were retrieved. We evaluated the strength of this relationship using odds ratios (ORs) with 95% confidence intervals. Sensitivity analysis was also conducted. RESULTS: A total of 24 individual studies were included in this meta-analysis. The risk of H. pylori infection was significantly lower in alcohol drinkers than nondrinkers (OR=0.83). People who drink wine (OR=0.90) or mixed types of alcoholic beverages (OR=0.78) had a lower risk of infection compared with those who drink beer. Among people aged 40 years or older, alcohol drinkers had a lower risk of H. pylori infection than nondrinkers (OR=0.68). Among people less than 40 years of age, alcohol drinking was not associated with H. pylori infection risk. Data showed that women were at a lower risk of H. pylori infection than men (OR=0.86). CONCLUSIONS: This meta-analysis suggests that the risk of H. pylori infection among alcohol drinkers is lower than that of nondrinkers. Drinking wine and mixed types of alcohol are better at reducing H. pylori infection than drinking beer. Nonetheless, we discourage reducing H. pylori infection through drinking, which increases the risk of other diseases.

Temperature-Shift-Induced Mechanical Property Evolution of Lithium-Ion Battery Separator Using Cyclic Nanoindentation.

The evolution of mechanical properties of separators affected by temperature shifts is imperative for the performance of the lithium-ion battery. The flexible film characteristics hinder the evaluation of the micromechanical properties of separators. In the present study, considering the susceptibility of separators to temperature fluctuations, the temperature distribution of the battery during the discharging process at subzero temperature is obtained. Three sets of separator samples subjected to various temperature shifts are prepared. Through multicycle depth-sensitive nanoindentation, the temperature-dependent weakening of elastic modulus and hardness of separators is experimentally verified. Moreover, the variation trends of elastic modulus, hardness, and hysteresis response of the separator specimens in terms of temperature are investigated via extracting from the multicycle loading-unloading nanoindentation responses. The temperature-dependent variations in the elastic modulus of the separator were investigated by following heating, cooling, and thermostatic processes. Meanwhile, the indentation tests also verify that the effect of temperature shifts on the hardness exhibits an attenuation trend when heating or cooling is followed by a thermostatic process. The variation analysis of nanoindentation hardness as a function of temperature shifts shows typical size effects dependent on the nanoindentation depth. The temperature-induced residual stress and elemental distribution are also analyzed through characterization using X-ray diffraction and energy-dispersive X-ray spectroscopy, respectively. The obtained evolution law of temperature shift-induced mechanical properties of a separator could facilitate the optimal design of the separators and provide the supporting data to enhance the safety performance of lithium-ion batteries.