Rituximab potentially improves clinical outcomes of CAR-T therapy for r/r B-ALL via sensitizing leukemia cells to CAR-T-mediated cytotoxicity and reducing CAR-T exhaustion.

PURPOSE: Despite chimeric antigen receptor (CAR) T-cell therapy has achieved great advances in recent year, approximately 50% of relapsed/refractory B cell acute lymphoblastic leukemia (r/r B-ALL) patients treated with CAR-T experience relapse 6 months post CAR-T treatment. CD20 express on 30 to 50% of B-ALL, which makes CD20 Monoclonal Antibody as one of the potential therapy strategies to decrease the tumor burden and improve the efficacy of CAR-T therapy. Adding Rituximab to chemotherapy protocol had been demonstrated to improve the outcome for CD20-positive ALL. However, rare study explored the influence of Rituximab combined with CAR-T therapy. METHODS: We retrospectively analyzed 20 r/r B-ALL patients who received CAR-T therapy, all of whom had failed multiple lines of therapy. Before CAR-T infusion, we administered Rituximab to 10 patients with high CD20 expression at a dose of 375 mg/m2 for 1 day. Meanwhile, we selected 10 patients with the comparable features who underwent CAR-T treatment without Rituximab in the same period as the control group. In vitro, the surface molecule expression and killing of CAR-T post Rituximab-treated B-ALL cells co-incubated with CAR-T cells were detected by flow cytometry. RESULTS: The median follow-up of Rituximab and Control groups were 29.27 and 9.83 months. We found that adding Rituximab may confer a favorable prognosis compared with Control group. The 2-year overall survival (OS) and leukemia-free survival (LFS) rates both were longer in the Rituximab group (90% vs. 26.7%, p = 0.0342; 41.7% vs. 25%, p = 0.308). In vitro, we observed that Rituximab-treated tumour cells are more sensitive to CAR-T killing and a broad range of cytokines and chemokines were produced when Rituximab-treated Nalm-6 cells co-cultured with 19-22CAR-T cells, such as interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2). To investigate whether Rituximab has an effect on CAR-T persistence, we stimulated CAR-T cells repeatedly in vitro with Rituximab-treated Nalm-6 to evaluate the changes in CAR-T surface exhaustion molecules at different times. We found that the expression of exhaustion molecules (LAG-3, PD-1, TIM-3) on CAR-T cells were significantly lower in the Rituximab group than in the Control group. CONCLUSION: Rituximab combined with CAR-T therapy is effective for improving the long-term prognosis of B-ALL patients who have failed multiple lines of therapy. In vitro, we observed that rituximab potentially improves CAR-T efficacy by sensitizing ALL to CART-mediated cytotoxicity and reducing CAR-T exhaustion.

Preparation and Application of Apatite-TiO2 Composite Opacifier: Preventing Titanium Glaze Yellowing through Pre-Combination.

In order to enhance the degree of binding reaction of TiO2 in titanium-containing ceramic glazes and prevent the reaction of its transformation into rutile to eliminate the yellowing phenomenon of the glaze surface, an apatite-TiO2 composite opacifier (ATO) was prepared through the mechanical grinding of hydroxyapatite and anatase TiO2. The properties, opacification mechanism, and yellowing inhibition of the prepared ceramic glazes were studied. The results show that the ATO is characterized by a uniform coating of TiO2 on the surface of the apatite and the formation of close chemical bonding between the apatite and TiO2. The ceramic glaze surface when using an ATO has a white appearance and excellent opacification performance. When an ATO was used, the L*, a*, and b* values of the glaze were 89.99, -0.85, and 3.37, respectively, which were comparable to those of a ZrSiO4 glaze (L*, a*, and b* were 88.24, -0.02, and 2.29, respectively). The opacification of the glaze was slightly lower than that of the TiO2 glaze (L* value was 92.13), but the appearance changed from yellow to the white of the TiO2 glaze (b* value was 9.18). The ceramic glaze layer when using an ATO mainly consists of titanite, glass phase, and a small amount of quartz, and the opacification mechanism is the crystallization of the generated titanite. ATOs can play an active role in solving the critical problem that arises when TiO2 replaces ZrSiO4 as an opacifier.

The clinical value of anal swabs for microbial detection in allogeneic haematopoietic stem cell transplantation.

The intestinal microbiota plays critical roles in allogeneic hematopoietic stem cell transplantation (allo-HSCT). Rapid and effective microbial detection methods have important guiding value for the selection of intervention strategies for allo-HSCT recipients. We evaluated the application of the anal swab test before transplantation in allo-HSCT recipients. A total of 120 patients who underwent anal swab testing before allo-HSCT were retrospectively analyzed and divided into 3 groups: sterile (aseptic growth-negative), G+ (gram-positive bacterial colonization), and G- (gram-negative bacterial colonization). On 16S rRNA sequencing, gram-negative bacteria predominated in the G- group before and after transplantation. Compared with the sterile group, the percentage of natural killer cells was higher and the percentage of T cells was lower after transplantation in the G- group at 1 month after transplantation. The percentage of CD4+ and CD4+CD8+ T cells was lower and the percentage of regulatory T cells was higher in the G- group. The plasma levels of proinflammatory cytokines (TNF-α, IFN-γ, IL-6, and IL-17A) at 2 weeks post-transplantation were lower in the G- group than in the sterile group, as was the cumulative incidence of grade III-IV acute graft-versus-host disease (GVHD). Gram-negative bacterial colonization before allo-HSCT was associated with low rates of bloodstream infections within 100 days post-transplantation and cytomegalovirus reactivation at 100 days to 2 years post-transplantation. Moreover, patients in the G- group had a higher rate of 2-year GVHD-free, relapse-free survival compared with patients in the sterile group. The detection results using anal swabs were consistent with the gram-negative or gram-positive bacteria abundance of 16S rRNA sequencing results and associated with immune homeostasis and clinical outcomes after allo-HSCT. Anal swab testing may have potential advantages as a simple and effective method for microbial detection in allo-HSCT.

Spatial distribution and potential sources of arsenic and water-soluble ions in the snow at Ili River Valley, China.

Trace elements and water-soluble ions in snow can be used as indicators to reveal natural and anthropogenic emissions. To understand the chemical composition, characteristics of snow and their potential sources in the Ili River Valley (IRV), snow samples were collected from 17 sites in the IRV from December 2018 to March 2019. Inverse distance weighting, enrichment factor (EF) analysis, and backward trajectory modelling were applied to evaluate the spatial distributions and sources of water-soluble ions and dissolved arsenic (As) in snow. The results indicate that Ca2+ and SO42- were the dominant ions, and the concentrations of As ranged from 0.09 to 0.503 µg L-1. High concentrations of As were distributed in the northwest and middle of the IRV, and the concentrations of the major ions were high in the west of the IRV. The strong correlation of As with F-, SO42-, and NO2- demonstrates that As mainly originated from coal-burning and agricultural activities. Principal component analysis showed that the ions originated from a combination of anthropogenic and crustal sources. The EFs showed that K+, SO42-, and Mg2+ were mainly influenced by human activities. Backward trajectory cluster analysis suggested that the chemical composition of snow was affected by soil dust transport from the western air mass, the unique terrain, and local anthropogenic activities. These results provide important scientific insights for atmospheric environmental management and agricultural production within the IRV.

Electrochemical Biosensor Based on Well-Dispersed Boron Nitride Colloidal Nanoparticles and DNA Aptamers for Ultrasensitive Detection of Carbendazim.

A selective electrochemical biosensor was developed for detecting carbendazim (CBZ) based on well-dispersed colloidal boron nitride (BN) nanocrystals and gold nanoparticles (Au NPs). BN was synthesized by "solvent cutting" to modify a glassy carbon electrode (GCE), and Au NPs were then electrodeposited. A single-stranded oligonucleotide with methylene blue (MB) was modified to the electrode surface through gold-sulfur bonds. A double-stranded DNA was formed in the presence of an aptamer. The aptamer chain can specifically bind to the target CBZ. When the aptamer binds to CBZ, the electroactive substance MB labeled at one end of the complementary chain can effectively contact the electrode surface. Detection of CBZ is realized by simultaneously monitoring the MB signal enhancement. The CBZ concentration was determined in a wide linearity range from 0.1 ng mL-1 to 100 µg mL-1, with a low detection limit of 0.019 ng mL-1. This biosensor exhibited excellent selectivity and acceptable repeatability and was applied in cucumber, kiwifruit, and water samples with good recoveries, demonstrating that the strategy has remarkable potential and offers a good platform for CBZ detection.

A Single-Chain Variable Fragment Antibody/Chemokine Fusion Protein Targeting Human Endoglin to Enhance the Anti-Tumor Activity of Cytokine-Induced Killer Cells.

Cytokine-induced killer cell immunotherapy is an ideal candidate for adoptive cell transfer therapy. However, therapeutic approaches to enhance the anti-tumor activity of cytokine-induced killer cells remain to be explored. Here, we described the successful development of a novel antibody-chemokine fusion protein containing the anti-human Endoglin antibody in the single-chain variable fragment format and human interferon-gamma-induced protein 10 (hENG scFv/hIP-10). Its anti-Endoglin immunoreactivity and chemotactic activity against the cytokine-induced killer cells were characterized in vitro. To evaluate the anti-tumor effect in vivo, cytokine-induced killer cells were intravenously injected into human hepatocellular carcinoma-bearing nude mice, together with intratumoral administration of the fusion protein hENG scFv/hIP-10 as an enhancer. The tumor volume and survival time of the mice were monitored, whilst the tumor-infiltrating cytokine-induced killer cells, serum levels of interferon-gamma, tumor cell proliferation, apoptosis, and angiogenesis were measured. The results demonstrated that hENG scFv/hIP-10 and cytokine-induced killer cells synergistically inhibited tumor growth and prolonged survival of tumor-bearing mice. Moreover, the number of tumor-infiltrating cytokine-induced killer cells, serum levels of interferon-gamma, and tumor cell apoptosis were increased, accompanied with decreased tumor proliferation and angiogenesis. Thus, our study suggests that hENG scFv/hIP-10 could enhance the anti-tumor activity of cytokine-induced killer cells against human hepatocellular carcinoma.

Tunable Adhesive Self-Cleaning Coating with Superhydrophobicity and Photocatalytic Activity.

Superhydrophobic coatings with intelligent properties have attracted much attention because of their wide application in many fields. However, there is a limited amount of literature on superhydrophobic coatings whose wettability and adhesion can be adjusted by UV irradiation and calcination at the same time. In this study, amorphous SiO2 microspheres (A-SiO2) and nano-TiO2 particles (N-TiO2) were used to fabricate A-SiO2/N-TiO2 composites by wet grinding, and then, they were modified with polydimethylsiloxane (PDMS) and sprayed onto substrate surfaces to obtain a tunable adhesive superhydrophobic A-SiO2/N-TiO2@PDMS coating. It is worth noting that the wettability and adhesion of the coating to water droplets could be adjusted by UV irradiation and calcination. The mechanisms of the aforementioned phenomena were studied. Moreover, methyl orange solution could be degraded by the coating due to its photocatalysis. The as-prepared coating had good adaptation to different substrates and outdoor environments. Moreover, the surfaces of these coatings exhibited the same liquid repellency towards different droplets. This research provides an environmental strategy to prepare advanced self-cleaning coatings.

Electrochemical aptasensor based on Mo2C/Mo2N and gold nanoparticles for determination of chlorpyrifos.

Two-dimensional Mo2C/Mo2N composites were synthesized by high temperature ball milling and used as support materials for fabricating a chlorpyrifos (CPF) aptasensor. Gold nanoparticles (Au NPs) were electrodeposited on the surface of a Mo2C/Mo2N-modified electrode to connect with the ferrocene (Fc) probe via Au-S bonds. The Fc probe can hybridize with the aptamer probe to form a double-stranded structure. The addition of CPF made the double strands melt and the Fc probe approached the surface of the electrode, thereby resulting in amplification of the electrochemical response. The current response of the aptasensor for detecting CPF in solutions linearly varied from 0 to 400 ng mL-1 (with a maximum at 0.98 V vs. Ag/AgCl). The Au NPs/Mo2C/Mo2N composites exhibited satisfactory electrochemical behavior due to their excellent electrical conductivity and large surface area. This ultrasensitive aptasensor showed a low limit of detection of 0.036 ng mL-1. It was applied to determine CPF in real samples with acceptable recoveries from 94.7 to 116.7%, and the relative standard deviation was from 2.57 to 7.08%.Graphical abstract Schematic diagram of the manufacturing process of the aptasensor. Electrochemical aptasensor based on Mo2C/Mo2N/Au NP composites show excellent performance in detecting CPF.

Recombinant Endoglin-Single-Chain Variable Fragment/ Induced Protein 10 Fusion Protein Potently Boosts the Anti-Tumor Efficacy of Adoptively Transferred TRP2-Specific CD8+ CD28+ Cytotoxic T Lymphocytes in Mice.

In this research, we studied the therapeutic efficacy of a newly designed fusion protein containing Endoglin single-chain variable fragment and IP10 (Endoglin-scFv/IP10), together with our recently generated TRP2-specific CD8+ CD28+ CTLs (CD8+ CD28+ CTLs) in controlling melanoma growth in mice. The recombinant Endoglin-scFv/IP10 was expressed in E. coli, purified by affinity chromatography, and characterized in vitro for its chemotactic movement and immunoreactivity with endoglin-expressing cells. In vivo, melanoma xenografts were established in mice (C57BL/6) using B16F10 cells. After that, mice were treated with intravenous injections of vehicle (PBS), Endoglin-scFv/IP10 alone, CD8+ CD28+ CTLs alone, or Endoglin-scFv/IP10+ CD8+ CD28+ CTLs. The therapeutic efficacy was assessed by monitoring tumor growth, mouse survival and cellular biomarkers. Endoglin-scFv/IP10 fusion protein combined with CD8+ CD28+ CTLs observed a reduction in tumor growth, resulting in improved survival. On the cellular level, the combination treatment dramatically reduced the number of systemic and tumor associated myeloid-derived suppressor cells or regulatory T cells, increased tumor-responsive interferon-γ-producing lymphocytes and tumor-associated CD8+ CXCR3+ T cells, and inhibited proliferation and angiogenesis but stimulated apoptosis within melanoma tissue. This study demonstrates the therapeutic potential of Endoglin-scFv/IP10 fusion protein in combination with CD8+ CD28+ CTLs in melanoma treatment.

Single-Domain Antibody-Based TCR-Like CAR-T: A Potential Cancer Therapy.

Retargeting the antigen-binding specificity of T cells to intracellular antigens that are degraded and presented on the tumor surface by engineering chimeric antigen receptor (CAR), also named TCR-like antibody CAR-T, remains limited. With the exception of the commercialized CD19 CAR-T for hematological malignancies and other CAR-T therapies aiming mostly at extracellular antigens achieving great success, the rareness and scarcity of TCR-like CAR-T therapies might be due to their current status and limitations. This review provides the probable optimized initiatives for improving TCR-like CAR-T reprogramming and discusses single-domain antibodies administered as an alternative to conventional scFvs and secreted by CAR-T cells, which might be of great value to the development of CAR-T immunotherapies for intracellular antigens.

One-Step Solution-Immersion Process of Hydrophobic Octyl Graphene Oxide-Modified Nickel Foam for Highly Efficient Oil-Water Separation.

Hydrophobic foam was prepared by immersing nickel foam in a dispersion of octyl group-grafted graphene oxide (GO) and used to clean up oil-water mixtures. Octyl graphene oxide (C8-GO) was synthesized using GO and triethoxyoctylsilane by a solvothermal method. The resulting coverage of the foam was characterized, and the surface morphology of the foam was also investigated. The static water contact angle (SWCA) was measured to evaluate the change in wettability of the developed foam. The pristine smooth microstructure of the nickel foam became rough after being covered with the C8-GO nanosheets. The SWCA of C8-GO nickel foam (C8-GO NF) surface was approximately 147°. The C8-GO NF can float on the surface of the water in contrast to the easy-sinking unmodified foam, demonstrating good hydrophobicity. Furthermore, the C8-GO NF showed outstanding performance in organic compound adsorption and excellent efficiency in oil-water separation. The reusability and durability of the obtained foam are evaluated to highlight its usability in more complicated scenarios. The use of C8-GO NF was proved to be a promising strategy for oil-water separation under harsh circumstance.

The diagnosis and prognosis values of WNT mRNA expression in colon adenocarcinoma.

WNT family genes have participated in the progression and development of many cancers, however, the association between colon adenocarcinoma (COAD) and WNTs have been rarely reported. This study investigated the significance of WNT genes expression in COAD from the standpoint of diagnosis and prognosis. The RNA-sequencing dataset of COAD was downloaded from The Cancer Genome Atlas and University of California, Santa Cruz Xena browser. The biology functions of WNT genes were investigated by biological analysis. Biological analysis of WNT family genes indicated that WNT genes were noticeably enriched in the complex process of WNT signaling pathway. The Pearson correlation analysis suggested WNT1 and WNT9B had a strong correlation. And receiver operating characteristic curves suggested that most of the genes could serve as a significant diagnostic makers in COAD (P < .05), especially WNT2 and WNT7B had high diagnostic values that the area under curve were 0.997 (95% confidence interval [0.994-1.000]) and 0.961 (95%CI [0.939-0.983]), respectively. And our multivariate survival analysis suggested the downregulated of WNT10B (P < .05) showed a favor prognosis in COAD overall survival. And the risk score model predicted that the upregulated expression of WNT10B might increase the risk of death. The very study we had conducted suggested that WNT genes had a certain connection with the diagnosis and prognosis of COAD. The messenger RNA expression of WNT2 and WNT7B might become potentially diagnostic biomarkers, and WNT10B might serve as an independent prognosis indicator for COAD.

Fabricated Electrochemical Sensory Platform Based on the Boron Nitride Ternary Nanocomposite Film Electrode for Paraquat Detection.

Hexagonal boron nitride (BN), an effective diffusion material for mass transport, was functionalized with molybdenum disulfide (MoS2) and Au nanoparticles (Au NPs). Then, the working electrodes with developed nanomaterials were applied to construct an electrochemical paraquat sensor. BN was prepared using a solid-state synthesis method combined with solvent-cutting. The electrochemical properties of the BN/MoS2/Au NP-based glassy carbon electrode (GCE) were investigated using differential pulse voltammetry and cyclic voltammetry. An excellent response signal to paraquat was found from 0.1 to 100 µM with a limit of detection of 0.074 µM, and it had acceptable reproducibility (relative standard deviation = 2.99%, n = 5) and good anti-interference ability. The modified GCE showed superior performance owing to the synergistic effects among all three given nanomaterials. With the proposed method, paraquat in grass samples from an orchard was then investigated. The results of the electrochemical analysis agreed with those of experiments and obtained a 96.28% confidence level via high-performance liquid chromatography, exhibiting relatively high stability. Therefore, the fabricated sensor can be a candidate for the determination of paraquat.