Photoluminescence-Electrochemiluminescence Dual-Mode Sensor Arrays for Histidine and Its Metabolite Discrimination and Disease Identification.

Histidine (His) and its metabolite analysis is significant due to their vital roles in the diagnosis of diseases. In practical applications, simple and effective detection and discrimination of these metabolic species are still a great challenge due to their highly similar structures. Herein, photoluminescence (PL)-electrochemiluminescence (ECL) dual-mode sensor arrays consisting of a series of sensing elements were proposed for simultaneous quantitation and accurate discrimination of His and its four key metabolites (including histamine, imidazole-4-acetic acid, N-acetylhistamine, and imidazole propionate). The sensing elements of these sensor arrays were constructed by employing two solvent iridium(III) complexes ([Ir(pbz)2(DMSO)Cl] and [Ir(ppy)2(DMSO)Cl], pbz = 3-(2-pyridyl)benzoic acid, ppy = 2-phenylpyridine) with excellent PL and ECL performances as cross-responsive sensing units. Based on diverse coordination abilities of the two complexes with the imidazole group of the five targets, PL and ECL responses of each sensing unit can be enhanced to various degrees, which generate unique fingerprint patterns for the corresponding targets. Through principal component analysis, the multifarious patterns (two-, three-, and four-element sensor arrays) can be transformed into simple visualization modes, from which His and its four key metabolites can be effectively discriminated against each other. Moreover, the quantitation of an individual metabolic species at different concentrations and the recognition of the mixtures with different ratios were also accurately achieved. Notably, His and its four key metabolites in urine can also be successfully discriminated by the as-fabricated sensor arrays, and the patients with kidney diseases can be identified clearly, providing a promising way for disease diagnosis.

Repair spinal cord injury with a versatile anti-oxidant and neural regenerative nanoplatform.

Spinal cord injury (SCI) often results in motor and sensory deficits, or even paralysis. Due to the role of the cascade reaction, the effect of excessive reactive oxygen species (ROS) in the early and middle stages of SCI severely damage neurons, and most antioxidants cannot consistently eliminate ROS at non-toxic doses, which leads to a huge compromise in antioxidant treatment of SCI. Selenium nanoparticles (SeNPs) have excellent ROS scavenging bioactivity, but the toxicity control problem limits the therapeutic window. Here, we propose a synergistic therapeutic strategy of SeNPs encapsulated by ZIF-8 (SeNPs@ZIF-8) to obtain synergistic ROS scavenging activity. Three different spatial structures of SeNPs@ZIF-8 were synthesized and coated with ferrostatin-1, a ferroptosis inhibitor (FSZ NPs), to achieve enhanced anti-oxidant and anti-ferroptosis activity without toxicity. FSZ NPs promoted the maintenance of mitochondrial homeostasis, thereby regulating the expression of inflammatory factors and promoting the polarization of macrophages into M2 phenotype. In addition, the FSZ NPs presented strong abilities to promote neuronal maturation and axon growth through activating the WNT4-dependent pathways, while prevented glial scar formation. The current study demonstrates the powerful and versatile bioactive functions of FSZ NPs for SCI treatment and offers inspiration for other neural injury diseases.

Inhibitory effect of pre-gelatinized dialdehyde starch on heat-induced deterioration of sea cucumber.

BACKGROUND: This study sought to investigate the inhibitory effect of pre-gelatinized dialdehyde starch (P-DAS) on the deterioration of sea cucumber during high-temperature sterilization. RESULTS: It was found that pre-gelatinization reduced crystallinity and average molecular weight of dialdehyde starch (DAS), exposed free aldehyde groups, improved the solubility, and unified the particle sizes. According to the texture profiles of sea cucumber, the crosslinking power of P-DAS was higher than that of DAS. The results of free amino content, total soluble substance, water retention, water distribution, relaxation time and scanning electron microscopy all showed that the crosslinking effect was dose-dependent on crosslinking agent. CONCLUSION: These results have proved that large molecules such as P-DAS, when properly handled, could also efficiently enter collagen hydrogels and perform crosslinking, providing reference for the development of new protein food stabilizing agents. © 2022 Society of Chemical Industry.

HE-YOLOv5s: Efficient Road Defect Detection Network.

In recent years, the number of traffic accidents caused by road defects has increased dramatically all over the world, and the repair and prevention of road defects is an urgent task. Researchers in different countries have proposed many models to deal with this task, but most of them are either highly accurate and slow in detection, or the accuracy is low and the detection speed is high. The accuracy and speed have achieved good results, but the generalization of the model to other datasets is poor. Given this, this paper takes YOLOv5s as a benchmark model and proposes an optimization model to solve the problem of road defect detection. First, we significantly reduce the parameters of the model by pruning the model and removing unimportant modules, propose an improved Spatial Pyramid Pooling-Fast (SPPF) module to improve the feature signature fusion ability, and finally add an attention module to focus on the key information. The activation function, sampling method, and other strategies were also replaced in this study. The test results on the Global Road Damage Detection Challenge (GRDDC) dataset show that the FPS of our proposed model is not only faster than the baseline model but also improves the MAP by 2.08%, and the size of this model is also reduced by 6.07 M.

IL-33 Released in the Liver Inhibits Tumor Growth via Promotion of CD4+ and CD8+ T Cell Responses in Hepatocellular Carcinoma.

IL-33 released by epithelial cells and immune cells functions as an alarmin and can induce both type 1 and type 2 immune responses. However, the role of IL-33 release in tumor development is still not clear. In this study, we examined the function of released IL-33 in murine hepatocellular carcinoma (HCC) models by hydrodynamically injecting either IL-33-expressing tumor cells or IL-33-expressing plasmids into the liver of tumor-bearing mice. Tumor growth was greatly inhibited by IL-33 release. This antitumor effect of IL-33 was dependent on suppression of tumorigenicity 2 (ST2) because it was diminished in ST2-/- mice. Moreover, HCC patients with high IL-33 expression have prolonged overall survival compared with the patients with low IL-33 expression. Further study showed that there were increased percentages and numbers of activated and effector CD4+ and CD8+ T cells in both spleen and liver in IL-33-expressing tumor-bearing mice. Moreover, IFN-γ production of the CD4+ and CD8+ T cells was upregulated in both spleen and liver by IL-33. The cytotoxicity of CTLs from IL-33-expressing mice was also enhanced. In vitro rIL-33 treatment could preferentially expand CD8+ T cells and promote CD4+ and CD8+ T cell activation and IFN-γ production. Depletion of CD4+ and CD8+ T cells diminished the antitumor activity of IL-33, suggesting that the antitumor function of released IL-33 was mediated by both CD4+ and CD8+ T cells. Taken together, we demonstrated in murine HCC models that IL-33 release could inhibit tumor development through its interaction with ST2 to promote antitumor CD4+ and CD8+ T cell responses.

IL-12/IL-18-preactivated donor NK cells enhance GVL effects and mitigate GvHD after allogeneic hematopoietic stem cell transplantation.

Adoptive transfer of donor NK cells has the potential of mediating graft-versus-leukemia (GVL) effect while suppressing acute graft-versus-host-disease (aGVHD) during allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, these beneficial effects are limited by the transient function of adoptively transferred NK cells. Previous studies demonstrate that cytokine-induced memory-like NK cells that are preactivated by IL-12, IL-15, and IL-18 have enhanced effector functions and long life span in vivo. Here, we investigated the effects of IL-12/18-preactivated and IL-12/15/18-preactivated donor NK cells on GVL and aGVHD in a murine model of allo-HSCT. We found that both IL-12/18- and IL-12/15/18-preactivated NK cells mediated stronger GVL effect than control NK cells mainly due to their elevated activation/cytotoxicity and sustained proliferative potential. Interestingly, we observed that although both IL-12/18- and IL-12/15/18-preactivated NK cells significantly inhibited severe aGVHD, only the IL-12/18-preactivated NK cells maintained the beneficial effect of donor NK cells on mild aGVHD. The IL-12/15/18-preactivated NK cell infusion accelerated aGVHD in the fully-mismatched mild aGVHD model. Our results demonstrated that IL-12/18-preactivated NK cells displayed sustained and enhanced GVL functions, and could mitigate aGVHD despite the severity of the disease. IL-12/18-preactivated donor NK cell infusion may be an effective and safe adoptive therapy after allo-HSCT.

Newcastle disease virus co-expressing interleukin 7 and interleukin 15 modified tumor cells as a vaccine for cancer immunotherapy.

Interleukin 15 (IL15) and IL7 are two cytokines essential for T cell development and homeostasis. In order to improve the antitumor activity by Newcastle disease virus (NDV)-modified tumor vaccine, we generated a recombinant NDV co-expressing IL15 and IL7 (LX/IL(15+7)) through incorporation of a 2A self-processing peptide into IL15 and IL7 using reverse genetics. B16 cells infected with LX/IL(15+7) expressed both IL15 and IL7 stably. The cytotoxicity assay showed that murine melanoma cells modified with LX/IL(15+7) could significantly enhance the antitumor immune response in vitro. Then, the antitumor effects of tumor vaccine modified with recombinant virus were tested in the murine tumor models. We observed strong antitumor responses induced by LX/IL(15+7)-modified tumor cells both in prophylaxis and therapeutic models. Although the tumor-infiltrating CD4+ T cells and CD8+ T cells were both increased, the antitumor activity of the tumor vaccine modified with LX/IL(15+7) was dependent on CD8+ T cells. Taken together, our data strongly indicated that tumor vaccine modified with NDV strain LX/IL(15+7) is a promising agent for cancer immunotherapy.

Brightly luminescent and color-tunable green-violet-emitting halide perovskite CH3NH3PbBr3 colloidal quantum dots: an alternative to lighting and display technology.

Organic-inorganic hybrid perovskite (CH3NH3PbX3, X = Cl, Br, or I) quantum dots have become one of the most promising materials for optoelectronic applications. We controllably synthesized CH3NH3PbBr3 quantum dots with a tunable spectrum with the emission peaks covering the range from green (523.6 nm), blue and eventually to deep violet (409.4 nm), which is wider than that of quantum dots obtained without changing the halide component. The mechanism of the blueshift was investigated. The purified quantum dots have allowed the fabrication of efficient electroluminescence devices having a simple glass/ITO/PEDOT:PSS/TFB/CH3NH3PbBr3 quantum dot/TPBi/LiF/Al structure. CH3NH3PbBr3 quantum dots with 5-30 µL n-octylamine showed an ideal color-saturated green emission with Commission Internationale de l'Eclairage color coordinates of (0.123, 0.744) and a narrow full width at half-maximum of 19-24 nm. The photoluminescence quantum yield was up to 90.2%. In addition, it is also worth noting that the chromaticity coordinates (x, y) of CH3NH3PbBr3 quantum dots with 50-100 µL n-octylamine are (0.300, 0.344), (0.305, 0.314) and (0.323, 0.318) in the white region. All these properties indicate that these MAPbBr3 quantum dots can provide effective data support for the application of white LEDs, and may potentially be used as single-component multicolor-emitting materials, which can be applied to lighting and display technology.

High quality factor photonic crystal filter at k ≈0 and its application for refractive index sensing.

We report here the refractive index (RI) sensing using the singly degenerate high quality factor (Q) modes in photonic crystal slabs (PCS) with the free-space coupled incident beam close to normal incidence. Q values of 3.2x104 and 1.8x104 were achieved for the fabricated PCS in air and aqueous solution, respectively. A spectral sensitivity (S) of 94.5 nm/RIU and a detection limit (DL) of 3x10-5 RIU were achieved with our device. Such a high-Q cavity for the singly degenerate mode close to normal incidence is very promising to achieve a lower DL for RI sensing.

Photonic crystal bandedge membrane lasers on silicon.

We report here the design and experimental demonstration of optically pumped photonic crystal bandedge membrane lasers on silicon-on-insulator (SOI) and on bulk silicon (Si) substrates, based on heterogeneously integrated InGaAsP multi-quantum-well membrane layers transfer printed onto patterned photonic crystal cavities. Single-mode lasing under room-temperature operation was observed at 1542 nm, with excellent side mode suppression ratio greater than 31.5 dB, for the laser built on SOI substrate. For the laser built on bulk Si substrate, single-mode lasing was also achieved at 1452 nm with much lower thermal resistance, as compared to that of the laser built on SOI substrates. Such improved thermal characteristics are favorable for lasers operating potentially at higher temperatures and higher power.

Optical Refractive Index Sensing Based on High-Q Bound States in the Continuum in Free-Space Coupled Photonic Crystal Slabs.

High sensitivity (S) and high quality factor (Q) are desirable to achieve low detection limit in label-free optical sensors. In this paper, we theoretically demonstrate that single-layer and coupled bi-layer photonic crystal slabs (PCS) possess simultaneously high S and high Q near the bound states in the continuum (BIC). We theoretically achieved S > 800 nm/RIU and Q > 107 in refractive index sensing in the 1400-1600 nm telecom optical wavelength bands. We experimentally demonstrated an S of 94 nm/RIU and a Q of 1.2 × 104, with a detection limit of 6 × 10-5 refractive index unit. These sensor designs can find applications in biochemical sensing, environmental monitoring, and healthcare.

Secreted IL-1α promotes T-cell activation and expansion of CD11b(+) Gr1(+) cells in carbon tetrachloride-induced liver injury in mice.

Interleukin-1α is mainly expressed on the cell membrane, but can also be secreted during inflammation. The roles of secreted and membrane IL-1α in acute liver inflammation are still not known. Here, we examined the functions of secreted and membrane IL-1α in a mouse model of carbon tetrachloride-induced acute liver injury. We show that secreted IL-1α aggravates liver damage and membrane IL-1α slightly protects mice from liver injury. Further studies showed that secreted IL-1α promotes T-cell activation. It also increased the expansion of CD11b(+) Gr1(+) myeloid cells, which may serve as a negative regulator of acute liver inflammation. Moreover, secreted IL-1α induced IL-6 production from hepatocytes. IL-6 neutralization reduced the proliferation of CD11b(+) Gr1(+) myeloid cells in vivo. CCL2 and CXCL5 expression was increased by secreted IL-1α in vitro and in vivo. Antagonists of the chemokine receptors for CCL2 and CXCL5 significantly reduced the migration of CD11b(+) Gr1(+) myeloid cells. These results demonstrate that secreted and membrane IL-1α play different roles in acute liver injury. Secreted IL-1α could promote T-cell activation and the recruitment and expansion of CD11b(+) Gr1(+) myeloid cells through induction of CCL2, CXCL5, and IL-6. The controlled release of IL-1α could be a critical regulator during acute liver inflammation.

[Study of low radiation exposure dose and low contrast medium dose in coronary CT angiography with High-pitch spiral acquisition mode of dual source CT].

OBJECTIVE: To evaluate the feasibility of low radiation exposure and low contrast medium volume for coronary CT angiography with High- pitch spiral acquisition mode of dual source CT. METHODS: 135 patients whose BMI <23 kg/m² and heart rates <65 bpm selected from 291 patients diagnosed of suspected CHD at our institution from September 2013 to February 2014 were randomly divided into 3 groups before CCTA, and there were 45 patients in each group. 80 kV , Iodixanol (320 mgI/ml) and sinogram affirmed iterative reconstruction (SAFIRE) were used in A group. 80 kV , Iopamidol (370 mgI/ml) and SAFIRE were used in B group. 100 kV, Iodixanol and filtered back projection (FBP) were used in C group. Two radiologists assessed image quality with 5-piont scale subjectively and double-blind. Independent-Sample Test was used to analyze statistical significance of image quality including signal to noise ratio(SNR) and contrast to noise ratio (CNR) between A and B group or between A and C group. At the same time, Contrast medium dose statistical significance between A and B group and mean effective Radiation dose (ED)statistical significance between A and C were analyzed by the same way. RESULTS: There were no significant difference of image quality including SNR and CNR of aortic root (AO), left main coronary artery (LM), left anterior descending artery (LAD), circumflex coronary artery (CX) and right coronary artery (RCA) Between A and B group (P = non-significant for all comparison), whereas Iodine in taken of A group decreased 14% (17 600 mg vs 20 350 mg). There were no significant difference of image quality including SNR and CNR of AO, LM, LAD, CX and RCA Between A and C group (P = non-significant for all comparison), whereas mean ED of A group decreased 50% (0.41 ± 0.05 mSv vs 0.79 ± 0.15 mSv). CONCLUSION: The double low dose application which use High-pitch spiral mode, 80 kV, SAFIRE, and Iodixanol (320 mgI/ml) can be used in those patients whose BMI <23 kg/m² and heart rates <65 bpm to reduce the burden of radiation and contrast medium significantly, without compromising the image quality.

B7-H3 enhances colorectal cancer progression by regulating HB-EGF via HIF-1α.

Background: B7-H3 (or CD276) represents an important costimulatory molecule expressed in many malignant solid tumors, including colorectal cancer (CRC). The receptor of B7-H3 is not known, and the intracellular function of B7-H3 remains obscure. Herein, we report that B7-H3 upregulated the epidermal growth factor heparin-binding epidermal growth factor (HB-EGF), likely by regulating hypoxia-inducible factor 1α (HIF-1α) and thereby promoting the progression of CRC. Methods: Lentiviral transfection was performed on CRC cells to establish stable low-B7-H3 expression cells. A mechanistic analysis with an Agilent human gene expression profiling chip was conducted on them. Clinical data and specimens were collected to detect the connection between B7-H3 and HB-EGF in CRC. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to detect the messenger RNA (mRNA) level of B7-H3, HB-EGF, and HIF-1α. Chromatin immunoprecipitation (ChIP) quantitative real-time PCR was conducted. The protein level of HIF-1α and the phosphatidylinositide 3-kinases (PI3K)-protein kinase B (AKT) pathway were detected by western blot. HIF-1α was recovered by lentiviral transfection, and the HB-EGF mRNA levels, proliferation, invasion, and angiogenesis ability were detected. Results: B7-H3 promoted tumor progression through HB-EGF and the PI3K-AKT pathway. As B7-H3 was downregulated, HB-EGF levels were significantly reduced simultaneously, a growth trend that was shown by both CRC cell lines and cancer tissues. In addition, B7-H3 and HB-EGF had significant associations with tumor-node-metastasis (TNM) stage and lymph node metastasis in 50 CRC patients. The binding ability of HIF-1α to the HB-EGF promoter region was significantly decreased in the shB7-H3 RKO group. Western blot revealed that PI3K, AKT, and mammalian target of rapamycin (mTOR) protein amounts and p-AKT and p-mTOR phosphorylation were also downregulated in shB7-H3 RKO cells, suggesting that B7-H3 may regulate HIF-1α via PI3K-AKT signaling. After recovery of the HIF-1α level by lentiviral transfection, the HB-EGF mRNA levels, proliferation, invasion, and angiogenesis in CRC cells recovered as well. Conclusions: B7-H3 may transmit intracellular signals through PI3K-AKT-mTOR-HIF-1α signaling, upregulating HB-EGF. As the final transcription factor of the pathway, HIF-1α regulates the transcription of the HB-EGF gene, thereby promoting HB-EGF expression, which eventually mediates cell proliferation, invasion, and angiogenesis and promotes the progression of CRC.

Determination of mine fault activation degree and the division of tectonic stress hazard zones.

This article conducts a comprehensive study on the activation characteristics of faults in the mine and analyzes the distribution patterns of the original rock stress field. Through quantitative research and analysis, we determine the partitioning characteristics of tectonic stress in the mine field under the dual effects of fault activation and original rock stress. The study also reveals the significant impact of different fault activation characteristics and different tectonic stress partitions on the stability of roadway surrounding rock. Using the Mohr-Coulomb strength criterion as a foundation, we investigate the mechanisms of fault activation and establish a mathematical model for fuzzy comprehensive evaluation. This model enables us to determine the strength level of fault activation in coal seam 9 of the Limin coal mine and construct a geological structure model. It has realized the transformation of fault activation degree from qualitative evaluation to quantitative evaluation. The stress state analysis software is used to draw the division of tectonic stress dangerous areas under the synergistic effect of fault activation and original rock stress. We then analyze the impact on the stability of roadway surrounding rock in these different hazardous areas. Utilizing the fuzzy comprehensive evaluation method, we take into account the impact of faults on the distribution characteristics of stress fields and the stability of roadway surrounding rock. This approach enables us to more accurately and comprehensively determine the hazardous areas of tectonic stress in the mine field under the dual effects of faults and original rock stress.

Application of dental pulp stem cells for bone regeneration.

Bone defects resulting from severe trauma, tumors, inflammation, and other factors are increasingly prevalent. Stem cell-based therapies have emerged as a promising alternative. Dental pulp stem cells (DPSCs), sourced from dental pulp, have garnered significant attention owing to their ready accessibility and minimal collection-associated risks. Ongoing investigations into DPSCs have revealed their potential to undergo osteogenic differentiation and their capacity to secrete a diverse array of ontogenetic components, such as extracellular vesicles and cell lysates. This comprehensive review article aims to provide an in-depth analysis of DPSCs and their secretory components, emphasizing extraction techniques and utilization while elucidating the intricate mechanisms governing bone regeneration. Furthermore, we explore the merits and demerits of cell and cell-free therapeutic modalities, as well as discuss the potential prospects, opportunities, and inherent challenges associated with DPSC therapy and cell-free therapies in the context of bone regeneration.

IL-37 dampens immunosuppressive functions of MDSCs via metabolic reprogramming in the tumor microenvironment.

Interleukin-37 (IL-37) has been shown to inhibit tumor growth in various cancer types. However, the immune regulatory function of IL-37 in the tumor microenvironment is unclear. Here, we established a human leukocyte antigen-I (HLA-I)-matched humanized patient-derived xenograft hepatocellular carcinoma (HCC) model and three murine orthotopic HCC models to study the function of IL-37 in the tumor microenvironment. We found that IL-37 inhibited HCC growth and promoted T cell activation. Further study revealed that IL-37 impaired the immunosuppressive capacity of myeloid-derived suppressor cells (MDSCs). Pretreatment of MDSCs with IL-37 before adoptive transfer attenuated their tumor-promoting function in HCC tumor-bearing mice. Moreover, IL-37 promoted both glycolysis and oxidative phosphorylation in MDSCs, resulting in the upregulation of ATP release, which impaired the immunosuppressive capacity of MDSCs. Collectively, we demonstrated that IL-37 inhibited tumor development through dampening MDSCs' immunosuppressive capacity in the tumor microenvironment via metabolic reprogramming, making it a promising target for future cancer immunotherapy.

Mine pressure behavior law of isolated island working face under extremely close goaf in shallow coal seam.

In order to study the mining pressure characteristics of the shallow buried coal seam with the same silo working face under the very close mining void zone and the overlying coal rock body, the theoretical analysis is used to determine whether the open-cutting eye bearing layer belongs to the mining under the very close mining void zone or not, based on the numerical simulation of FLAC3D and on-site measurement of the working resistance at the end of the cycle of the working face's hydraulic bracket, It is proposed to divide the mining stress of the working face based on the advancing length of the working face, that is, the high-pressure zone, the transition zone and the low-pressure zone. The results of the study show that: FLAC3D software was used to analyze the stress intensity of the "C" island working face when it was mined back to 50 m, 100 m, 150 m, and 180 m (one time "square"), and the simulation results were imported into the Origin software, which was used to analyze the stress intensity of the working face. The simulation results were imported into Origin software, and the influence range of mining stress was divided into four areas: high-stress area, stress transition area, low-stress area, and "square" stress concentration area. According to the on-site measurement of the working resistance at the end of hydraulic support cycle, the initial pressure step of the working face under the overlying coal rock body is 48.9-55.7 m, with the peak value of 38 MPa, the cycle pressure step is 9.0-23.3 m, with the peak value of 36 MPa, and the dynamic load factor of the working face is 1.14-1.16; relative to the overlying coal rock body, the average decrease of the cycle pressure step is nearly 10% and the average increase of dynamic load factor is 1.14-1.16; compared with that under the overlying coal rock body, the average decrease of the cycle pressure step is nearly 10% and the average increase of dynamic load factor is 1.14-1.16. Compared with the overlying coal rock body, the average decrease of the cycle pressure step under the overlying mining zone is nearly 10%, the average increase of the dynamic load factor is 20%, and there is no obvious regularity and periodicity in the direction of strike, and the working face is divided into three parts along the direction of strike: high-pressure zone, transition zone, and low-pressure zone. Therefore, in the process of mining under the overlying coal rock body, we should strengthen the roadway peripheral rock support and roof and floor management, which is of guiding significance to the mining of similar working faces and mine safety production.

Oxidized inositol stabilizes rehydrated sea cucumbers against non-enzymatic deterioration.

Rehydrated sea cucumber (RSC) is a popular seafood in northeast Asia with high deep-processing potential. Yet the development of value-added RSC product is limited by non-enzymatic decrosslinking of RSC collagen hydrogel system, which could potentially be solved by crosslinking enhancement. In this work, NaIO4 oxidized inositol (NOXIN), a novel crosslinker, was synthesized and its capacity in RSC stabilization was evaluated with texture profile analysis, free amino group content, total soluble protein, water activity, water withholding capacity and scanning electron microscope (SEM). The results show that NOXIN could stabilize RSC in a dose dependent manner by crosslinking the amino groups on collagen, thus enhances the mechanical strength and water holding capacity, retains water activity, and suppresses protein degradation of the hydrogel system, although overdosing compromises the stabilizing effect. The results of this study have not only validated the capacity of NOXIN as protein crosslinker, but also provided reference to the development of new crosslinkers.

IL12/18/21 Preactivation Enhances the Antitumor Efficacy of Expanded γδT Cells and Overcomes Resistance to Anti-PD-L1 Treatment.

γδT cells are promising candidates for cellular immunotherapy due to their immune regulation through cytokine production and MHC-independent direct cytotoxicity against a broad spectrum of tumors. However, current γδT cell-based cancer immunotherapy has limited efficacy, and novel strategies are needed to improve clinical outcomes. Here, we report that cytokine pretreatment with IL12/18, IL12/15/18, IL12/18/21, and IL12/15/18/21 effectively enhanced the activation and cytotoxicity of in vitro-expanded murine and human γδT cells. However, only adoptive transfer of IL12/18/21 preactivated γδT cells significantly inhibited tumor growth in a murine melanoma model and a hepatocellular carcinoma model. Both IL12/18/21 preactivated antibody-expanded and zoledronate-expanded human γδT cells effectively controlled tumor growth in a humanized mouse model. IL12/18/21 preactivation promoted γδT cell proliferation and cytokine production in vivo and enhanced IFNγ production and activation of endogenous CD8+ T cells in a cell-cell contact- and ICAM-1-dependent manner. Furthermore, the adoptive transfer of IL12/18/21 preactivated γδT cells could overcome the resistance to anti-PD-L1 therapy, and the combination therapy had a synergistic effect on the therapeutic outcomes. Moreover, the enhanced antitumor function of adoptively transferred IL12/18/21 preactivated γδT cells was largely diminished in the absence of endogenous CD8+ T cells when administered alone or in combination with anti-PD-L1, suggesting a CD8+ T cell-dependent mechanism. Taken together, IL12/18/21 preactivation can promote γδT cell antitumor function and overcome the resistance to checkpoint blockade therapy, indicating an effective combinational cancer immunotherapeutic strategy.