Injectable hydrogels activated with copper sulfide nanoparticles for enhancing spatiotemporal sterilization and osteogenesis in periodontal therapy.

Developing biomaterials capable of promoting bone regeneration in bacteria-infected sites is of utmost urgency for periodontal disease therapies. Here we produce a hybrid hydrogel by integrating CuS nanoparticles (CuSNPs), which could kill bacteria through photothermal therapy (PTT) triggered by a near infrared (NIR) light, and a gelatin methacryloyl (GelMA) hydrogel, which is injectable and biocompatible. Specifically, CuSNPs were precipitated by chitosan (CS) firstly, then grafted with methacrylic anhydride (MA) to form CuSNP@CS-MA, which was photo-crosslinked with GelMA to synthesize hybrid hydrogels (GelMA/CuSNP). The hybrid hydrogels exhibited a broad-spectrum antibacterial property that could be spatiotemprorally manipulated through applying a NIR light. Their mechanical properties were adjustable by controlling the concentration of CuSNPs, enabling the hydrogels to become more adapted to the oral diseases. Meanwhile, the hybrid hydrogels showed good cytocompatibility in vitro and improved hemostasis in vivo. Moreover, they accelerated alveolar osteogenesis and vascular genesis, successfully treating periodontis in four weeks in a rat model. GelMA/CuSNP hydrogels showed a broad-spectrum sterilization ability via PTT in vitro and outstanding antibacterial property in vivo, suggesting that the hybrid hydrogels could function in the challenging, bacteria-rich, oral environment. Such injectable hybrid hydrogels, capable of achieving both facilitated osteogenesis and NIR-inducible sterilization, represent a new biomaterial for treating periodontitis.

Real-time and quantitative protein detection via polyacrylamide gel electrophoresis and online intrinsic fluorescence imaging.

The detection of intrinsic protein fluorescence is a powerful tool for studying proteins in their native state. Thanks to its label-free and stain-free feature, intrinsic fluorescence detection has been introduced to polyacrylamide gel electrophoresis (PAGE), a fundamental and ubiquitous protein analysis technique, to avoid the tedious detection process. However, the reported methods of intrinsic fluorescence detection were incompatible with online PAGE detection or standard slab gel. Here, we fulfilled online intrinsic fluorescence imaging (IFI) of the standard slab gel to develop a PAGE-IFI method for real-time and quantitative protein detection. To do so, we comprehensively investigated the arrangement of the deep-UV light source to obtain a large imaging area compatible with the standard slab gel, and then designed a semi-open gel electrophoresis apparatus (GEA) to scaffold the gel for the online UV irradiation and IFI with low background noise. Thus, we achieved real-time monitoring of the protein migration, which enabled us to determine the optimal endpoint of PAGE run to improve the sensitivity of IFI. Moreover, online IFI circumvented the broadening of protein bands to enhance the separation resolution. Because of the low background noise and the optimized endpoint, we showcased the quantitative detection of bovine serum albumin (BSA) with a limit of detection (LOD) of 20 ng. The standard slab gel provided a high sample loading volume that allowed us to attain a wide linear range of 0.03-10 µg. These results indicate that the PAGE-IFI method can be a promising alternative to conventional PAGE and can be widely used in molecular biology labs.

A facile double moving redox boundary model for visual electrophoresis titration of ascorbic acid.

In this work, we proposed a double moving redox boundary (MROB) model to realize the colorless analyte electrophoresis titration (ET) by the two steps of the redox reaction. Single MROB has been proposed for the development of ET sensing (Analyst, 2013, 138, 1137. ACS Sensor, 2019, 4, 126.), and faces great challenges in detecting the analyte without color change during redox reaction. Herein, a novel model of double-MROB electrophoresis, including its mechanisms, equations, and procedures, was developed for titration by using ascorbic acid as a model analyte. The first MROB was created with ferric iron (Fe3+) and iodide ion (I-) in which Fe3+ was reduced as Fe2+ and I- was oxidized as molecular iodine (I2) used as an indicator of visible MROB due to blue starch-iodine complex. The second boundary was then formed between the molecular iodine and model analyte of ascorbic acid. Under given conditions, there was a quantitative relationship between velocity of MROB (VMROB(ii)) and ascorbic acid concentration (CVit C) in the double-MROB system (1/VMROB(ii) = 0.6502CVit C + 4.5165, and R = 0.9939). The relevant relative standard deviation values of intraday and inter-day were less than ∼5.55% and ∼6.64%, respectively. Finally, the titration of ascorbic acid in chewable vitamin C tablets was performed by the developed method, the titration results agreed with those via the classic iodometric titration. All the results briefly demonstrated the validity of the double MROB model, in which Vit C was used as a model analyte. The developed method had potential use in quantitative analysis of redox-active species in biomedical samples.

Nondestructive and high-resolution monitoring of inflammation-type skull defects regeneration on adult zebrafish with optical coherence tomography.

Optimized animal models and effective imaging techniques are exceedingly important to study cranial defects in bone loss due to chronic inflammation. In this study, the assessment procedure on a zebrafish inflammation-type skull defects model was monitored in vivo with spectral-domain optical coherence tomography (SD-OCT), and the efficacy of etidronate disodium in bone regeneration was assessed. An acute skull defect injury model was established in adult zebrafish using a stereotaxic craniotomy device. SD-OCT imaging was performed immediately following the mechanical injury. Both SD-OCT and immunohistochemistry results demonstrated an increase in inflammation-induced skull destruction within 5 days, which was confirmed by pathological experiments.

Marker-Free Isoelectric Focusing Patterns for Identification of Meat Samples via Deep Learning.

Isoelectric focusing (IEF) is a powerful tool for resolving complex protein samples, which generates IEF patterns consisting of multiplex analyte bands. However, the interpretation of IEF patterns requires the careful selection of isoelectric point (pI) markers for profiling the pH gradient and a trivial process of pI labeling, resulting in low IEF efficiency. Here, we for the first time proposed a marker-free IEF method for the efficient and accurate classification of IEF patterns by using a convolutional neural network (CNN) model. To verify our method, we identified 21 meat samples whose IEF patterns comprised different bands of meat hemoglobin, myoglobin, and their oxygen-binding variants but no pI marker. Thanks to the high throughput and short assay time of the microstrip IEF, we efficiently collected 1449 IEF patterns to construct the data set for model training. Despite the absence of pI markers, we experimentally introduced the severe pH gradient drift into 189 IEF patterns in the data set, thereby omitting the need for profiling the pH gradient. To enhance the model robustness, we further employed data augmentation during the model training to mimic pH gradient drift. With the advantages of simple preprocessing, a rapid inference of 50 ms, and a high accuracy of 97.1%, the CNN model outperformed the traditional algorithm for simultaneously identifying meat species and cuts of meat of 105 IEF patterns, suggesting its great potential of being combined with microstrip IEF for large-scale IEF analyses of complicated protein samples.

A handheld contactless conductivity detector for monitoring the desalting of low-volume virus and cell samples.

Desalting of biosamples is crucial for analytical techniques intolerant to abundant salts. However, there is no simple tool to monitor the desalting of low-volume biosamples so far. Here we developed a handheld capacitively coupled contactless conductivity detector (hC4D) as a miniaturized device to measure the conductivity of 75 µL biosamples. Polyether-ether-ketone (PEEK) tubing was selected as the sample reservoir for sample loading via a pipette. Another pipetting of air pushed the sample solution out of the tubing to recollect the sample. Owing to the low sample consumption and easy sample recollection, hC4D is advantageous for testing expensive biosamples, such as viruses and cells. In addition, the whole process of sample injection, conductivity measurement, recollection, and calibration of conductivity can be completed within 1 min. To verify the feasibility of hC4D, we monitored the desalting progress of gel filtration (GF) of 200 µL blood samples, ultrafiltration (UF) of 300 µL virus samples, and dialysis of 7 mL cell samples. Three rounds of GF and UF completely removed the salts but led to poor sample recovery. In contrast, low concentrations of residual salts remained and better recovery was achieved after two rounds of GF and UF. We further utilized the hC4D to monitor the dialysis and tuned the salt concentration in the cell sample, such that we maintained the viability of cells in a low conductivity environment. These results indicated that hC4D is a promising tool for optimizing the desalting procedure of low-volume biosamples.

Transcriptomic landscape of GC-2spd(ts) cell in response to the downregulation of piR-1207/2107.

Spermatogenesis is a highly orchestrated dynamic developmental process, during which piRNAs play an indispensable role. Our previous studies have confirmed that the levels of piR-1207 and piR-2107 were significantly decreased in spermatozoa and seminal plasma of patients with asthenozoospermia, compared with the fertile controls. In order to explore the function of piR-1207 and piR-2107 in human spermatogenesis and their potential regulatory downstream genes, we examined the transcriptomic landscape in mouse spermatocyte cell line GC-2spd(ts) transfected with anti-piR-1207 and anti-piR-2107 by RNA sequencing. The result showed that 86 and 75 differential expression genes (DEGs) in anti-piR-1207 and anti-piR-2107 group, respectively. Among the DEGs, three genes including Pbp2, Pde3a and Cage1 were identified as potential key genes playing important roles in mediating sperm motility and morphology in anti-piR-1207 or anti-piR-2107 transfected transcriptomic response. Next, the expression levels of Pbp2, Pde3a and Cage1 were confirmed by qRT-PCR. These results showed that Pbp2, Pde3a and Cage1 may serve as the potential regulatory genes of piR-1207 or piR-2107. In summary, piR-1207 and piR-2107 might have an implication in modulating the process of spermatogenesis through regulating the expression of potential genes.

Chinese adult segmentation according to health skills and analysis of their use for smart home: a cross-sectional national survey.

BACKGROUND: Digital health has become a heated topic today and smart homes have received much attention as an important area of digital health. Smart home is a device that enables automation and remote control in a home environment via the internet. However, most of the existing studies have focused on discussing the impact of smart home on people. Only few studies have focused on relationship between health skills and use of smart home. AIMS: To analyze the health skills of Chinese adults and segment them to compare and analyze the use of smart home for each group. METHODS: We used data from 11,031 participants aged 18 and above. The population was clustered based on five health skills factors: perceived social support, family health, health literacy, media use, and chronic diseases self-behavioral management. A total of 23 smart homes were categorized into three sub-categories based on their functions: entertainment smart home, functional smart home, and health smart home. We analyzed demographic characteristics and utilization rate of smart home across different cluster. Each groups' features and the differences in their needs for smart home functions were compared and analyzed. RESULTS: As a result of the survey on health skills, three groups with different characteristics were clustered: good health skills, middle health skills, and poor health skills. The utilization rate of smart home was the highest was good health skills group (total smart home: 92.7%; entertainment smart home: 61.1%, functional smart home: 77.4%, and health smart home: 75.3%; P < 0.001). For entertainment smart home, smart TV had the highest utilization rate (good health skills: 45.7%; middle health skills: 43.5%, poor health skills: 33.4%, P < 0.001). For functional smart home, smart washing machine (good health skills: 37.7%, middle health skills: 35.11%, poor health skills: 26.5%; P < 0.001) and smart air conditioner (good health skills: 36.0%, middle health skills: 29.1%, poor health skills: 24.6%) were higher than other of this category. For health smart home, sports bracelet has the highest utilization rate (good health skills: 37.3%, middle health skills: 24.5%, poor health skills: 22.8%). CONCLUSION: People can be divided into different categories based on health skill profiles, those with good health skills had a better utilization rate of smart home. The government and smart home companies need to focus on people with poor smart home use in various ways to promote their use of smart homes for personal health management.

Complex and Multilayered Role of IL-21 Signaling during Thymic Development.

Unlike IL-7, which is known to be critical for T cell thymic development, the role of IL-21 in this process is still controversial. IL-21 has been shown to accelerate thymic recovery in mice treated with glucocorticoids and revives the peripheral T cell pool in aged animals. However, mice with a defect in IL-21 signaling exhibit normal thymic cellularity, challenging the importance of this cytokine in the thymic developmental process. Using mixed bone marrow chimeric mice, our studies describe a multilayered role for IL-21 in thymopoiesis. In this system, IL-21R-deficient cells are unable to compete with wild-type populations at different stages of the thymic development. Using a mixed bone marrow chimeric animal model, IL-21 seems to be involved as early as the double-negative 1 stage, and the cells from the knockout compartment have problems transitioning to subsequent double-negative stages. Also, similar to IL-7, IL-21 seems to be involved in the positive selection of double-positive lymphocytes and appears to play a role in the migration of single-positive T cells to the periphery. Although not as critical as IL-7, based on our studies, IL-21 plays an important complementary role in thymic T cell development, which, to date, has been underrecognized.

Toxoplasma: Immunity and Pathogenesis.

Toxoplasma gondii infection induces a strong immunity in the host. Although the response is manifested by innate response during early infection, adaptive immunity is critical for long-term protection. Amongst the adaptive immune response CD4 T cells play an important helper role for CD8 T cells which are the primary effector cells responsible for controlling the infection. Notwithstanding the induction of robust CD8 T immunity during acute infection, the parasite is not eradicated. One of the reasons for this is the functional exhaustion of CD8 T cells during latent infection. Recent studies from our laboratory have reported that primary cause of CD8 T cell exhaustion is compromised CD4 T cell help during latent toxoplasmosis. CD8 T cell dysfunctionality is preceded by CD4 exhaustion and effector immunity is severely compromised.