Autologous Nanobody-Derived Fratricide-Resistant CD7-CAR T-cell Therapy for Patients with Relapsed and Refractory T-cell Acute Lymphoblastic Leukemia/Lymphoma.

PURPOSE: Since CD7 may represent a potent target for T-lymphoblastic leukemia/lymphoma (T-ALL/LBL) immunotherapy, this study aimed to investigate safety and efficacy of autologous CD7-chimeric antigen receptor (CAR) T cells in patients with relapsed and refractory (R/R) T-ALL/LBL, as well as its manufacturing feasibility. PATIENTS AND METHODS: Preclinical phase was conducted in NPG mice injected with Luc+ GFP+CCRF-CEM cells. Open-label phase I clinical trial (NCT04004637) enrolled patients with R/R CD7-positive T-ALL/LBL who received autologous CD7-CAR T-cell infusion. Primary endpoint was safety; secondary endpoints included efficacy and pharmacokinetic and pharmacodynamic parameters. RESULTS: CD7 blockade strategy was developed using tandem CD7 nanobody VHH6 coupled with an endoplasmic reticulum/Golgi-retention motif peptide to intracellularly fasten CD7 molecules. In preclinical phase CD7 blockade CAR T cells prevented fratricide and exerted potent cytolytic activity, significantly relieving leukemia progression and prolonged the median survival of mice. In clinical phase, the complete remission (CR) rate was 87.5% (7/8) 3 months after CAR T-cell infusion; 1 patient with leukemia achieved minimal residual disease-negative CR and 1 patient with lymphoma achieved CR for more than 12 months. Majority of patients (87.5%) only had grade 1 or 2 cytokine release syndrome with no T-cell hypoplasia or any neurologic toxicities observed. The median maximum concentration of CAR T cells was 857.2 cells/µL at approximately 12 days and remained detectable up to 270 days. CONCLUSIONS: Autologous nanobody-derived fratricide-resistant CD7-CAR T cells demonstrated a promising and durable antitumor response in R/R T-ALL/LBL with tolerable toxicity, warranting further studies in highly aggressive CD7-positive malignancies.

Cu-doped g-C3N4 catalyst with stable Cu0 and Cu+ for enhanced amoxicillin degradation by heterogeneous electro-Fenton process at neutral pH.

The development of new heterogeneous Cu-based solid catalysts for hydroxyl radical (∙OH) generation plays a crucial role in degradation of pollutants at neutral pH circumstance. In this work, a Cu-doped graphitic carbon nitride (g-C3N4) complex was synthesized in one-step pyrolysis process using copper chloride dihydrate and dicyandiamide as precursors. The results reveal that after Cu doping, the bulk structure of g-C3N4 was destroyed with fragmentary morphology formation. Besides, Cu0 and Cu+ were successfully embedded in g-C3N4 sheet. Moreover, amoxicillin (AMX) removal by heterogeneous electro-Fenton process was performed to evaluate the catalytic activity of the Cu-doped g-C3N4. 99.1% AMX removal efficiency was obtained after 60 min electrolysis under neutral pH condition when the current density was 12 mA cm2 and the catalyst dosage was 0.3 g L-1. Both Cu0 and Cu+ were stably retained in the Cu-doped g-C3N4 catalyst and AMX removal efficiency reached 91.1%, even after 5 cycles, manifesting the remarkable stability of Cu-doped g-C3N4. Also, Cu-doped g-C3N4 possessed excellent catalytic activities for AMX removal in various waterbodies. According to the catalytic mechanism analysis, the ∙OH was proved to be the primary reactive species for AMX removal in heterogeneous electro-Fenton process. Based on the identification of sixteen different intermediate products, the possible degradation pathways were proposed. This work provides a simple method to synthesize a Cu-based solid catalyst containing stable Cu0 and Cu + for degradation of pollutants in wastewater.

[Killing Effect of A CD7 Chimeric Antigen Receptor-Modified NK-92MI Cell Line on CD7-Positive Hematological Malignant Cells].

OBJECTIVE: To investigate the killing effect of NK-92MI cells modified by chimeric antigen receptor (CD7-CAR) and specifically targeting CD7 to CD7+ hematological malignant cells. METHODS: Three types of hematological malignant tumor cells, including 5 cases of CD7+ acute T-lymphoblastic leukemia (T-ALL), 10 cases of acute myeloid leukemia (AML) and 6 cases of T-cell lymphoma were collected, centrifuged, cultured and used to detect the expression levels of tumor cell surface targets; 7-AAD, CD56-APC, CD3-FITC, IgG Fc-PE flow cytometry were used to detected the transfection efficiency of NK-92MI and CD7-CAR-NK-92MI cells, killing efficiencies of CD7-CAR-NK-92MI cells to CD7+ hematological tumor cells in vitro were determined by flow cytometry using PE Annexin V Apoptosis Detection Kit. Secretion differences of NK-92MI and CD7-CAR-NK-92MI cytokines interleukin (IL)-2, interferon (IFN)-γ, and granzyme B detection were estimated by using CBA kit. RESULTS: The killing efficiencies of CD7-CAR-modified NK-92MI cells to CD7+ T-ALL, AML, T-cell lymphoma tumor cells were significantly higher than those of NK-92MI cells without genetical modification. The difference showed statistically significant (P＜0.05). The level of IFN-γ and granzyme B were significantly increased among cytokines secreted by CD7-CAR-modified NK-92MI cells as compared with those of NK-92MI cells without genetical modification (P＜0.05) . CONCLUSION: CD7-CAR-modified NK-92MI cells have significantly improved killing efficiency against CD7+ T-ALL, AML and T lymphoma cells, and shows specific targeting effects, which provides a clinical basis for the treatment of CD7+ hematological malignancies.

A hybrid prediction model for forecasting wind energy resources.

Wind energy is important to the transformation and development of global energy, because it is clean and renewable. However, the productivity of wind power is low due to its volatility, randomness, and uncertainty. Therefore, a new hybrid prediction model based on combined Elman-radial basis function (RBF) and Lorenz disturbance is proposed, which can promote the productivity of wind power by better predicting wind speed, firstly, applying the variational mode decomposition (VMD) algorithm to original nonstationary wind speed data to obtain several relatively stationary intrinsic mode functions (IMF), so as to fully exploit its potential characteristics. Meanwhile, the sample entropy is introduced to determine the decomposition number K. Afterwards, different IMF components with different characteristics are used for training and prediction: Elman neural network with sensitivity to historical state data is used for wind speed trend components; RBF with strong nonlinear mapping capability is adopted for other stochastic modal components. Next, the first-step prediction values can be obtained by reconstructing the predicted results of the respective IMF components. Finally, the Lorenz equation is introduced in view of the effects of atmospheric disturbances on wind fluctuations, which can be used to revise the first-step prediction results to obtain more realistic prediction results. By experimenting with the real data from two different wind farms and comparing with other predictive models, we found that (1) VMD can solve the problem of modal aliasing in empirical mode decomposition, to obtain a better decomposition result; (2) the combined prediction method of Elman and RBF is used for modal components, that is, different algorithms are adopted for different components, which have better prediction effects; (3) in this research, the results of the proposed combination prediction model is more accurate by comparison with the other neural network models. This research work will help the power system to rationally formulate wind farm control strategies, enhance the self-regulation of wind farm, and further promote global energy innovation.

Phytochemical and comparative transcriptome analyses reveal different regulatory mechanisms in the terpenoid biosynthesis pathways between Matricaria recutita L. and Chamaemelum nobile L.

BACKGROUND: Matricaria recutita (German chamomile) and Chamaemelum nobile (Roman chamomile) belong to the botanical family Asteraceae. These two herbs are not only morphologically distinguishable, but their secondary metabolites - especially the essential oils present in flowers are also different, especially the terpenoids. The aim of this project was to preliminarily identify regulatory mechanisms in the terpenoid biosynthetic pathways that differ between German and Roman chamomile by performing comparative transcriptomic and metabolomic analyses. RESULTS: We determined the content of essential oils in disk florets and ray florets in these two chamomile species, and found that the terpenoid content in flowers of German chamomile is greater than that of Roman chamomile. In addition, a comparative RNA-seq analysis of German and Roman chamomile showed that 54% of genes shared > 75% sequence identity between the two species. In particular, more highly expressed DEGs (differentially expressed genes) and TF (transcription factor) genes, different regulation of CYPs (cytochrome P450 enzymes), and rapid evolution of downstream genes in the terpenoid biosynthetic pathway of German chamomile could be the main reasons to explain the differences in the types and levels of terpenoid compounds in these two species. In addition, a phylogenetic tree constructed from single copy genes showed that German chamomile and Roman chamomile are closely related to Chrysanthemum nankingense. CONCLUSION: This work provides the first insights into terpenoid biosynthesis in two species of chamomile. The candidate unigenes related to terpenoid biosynthesis will be important in molecular breeding approaches to modulate the essential oil composition of Matricaria recutita and Chamaemelum nobile.

Fabrication of multi-walled carbon nanotubes and carbon black co-modified graphite felt cathode for amoxicillin removal by electrochemical advanced oxidation processes under mild pH condition.

Hydrogen peroxide (H2O2) electrogenerated via two-electron oxygen reduction reaction at cathode plays an important role in electrochemical advanced oxidation processes for organic pollutants removal from wastewater. Herein, multi-walled carbon nanotubes and carbon black co-modified graphite felt electrode (MWCNTs-CB/GF) was prepared as an efficient cathode for H2O2 electrogeneration and amoxicillin removal by anodic oxidation with hydrogen peroxide (AO-H2O2) and electro-Fenton (EF) under mild pH condition. Besides, the physicochemical and electrochemical properties of MWCNTs-CB/GF were characterized by scanning electron microscopy, N2 adsorption and desorption experiment, contact angle measurement, X-ray photoelectron spectroscopy, and linear sweep voltammetry. Compared with GF, MWCNTs-CB/GF showed a higher H2O2 generation of 309.0 mg L-1 with a current efficiency of 60.9% (after 120 min) and more effective amoxicillin removal efficiencies of 97.5% (after 120 min) and 98.7% (after 30 min) in AO-H2O2 and EF (with 0.5 mM Fe2+) processes, under the condition of current density 12 mA cm-2 and initial pH 5.5. Meanwhile, the TOC removal efficiency was 45.2% during EF process after 120 min. Anodic oxidation, H2O2 oxidation, and methanol capture indicated that ∙OH generated via electro-activation reaction at MWCNTs-CB/GF and Fenton reaction in solution played the dominant role in amoxicillin removal. Moreover, the TOC removal was associated with ∙OH generated during Fenton reaction in the solution. The major intermediates of AMX degradation by EF process were identified using LC-MS and the possible degradation pathways were proposed containing of ß-lactam ring opening, hydroxylation reaction, decarboxylation reaction, methyl groups in the thiazolidine ring oxidation reaction, bond cleavage, and rearrangement processes. All of the above results proved that MWCNTs-CB/GF was an excellent cathode for AMX degradation under mild pH condition.

Immune-enhancing effects of polysaccharides extracted from Lilium lancifolium Thunb.

Lilium lancifolium Thunb. is a Chinese traditional plant with various health benefits. In this study, we purified and characterized the water-soluble polysaccharide fraction (LLP-1A) of L. lancifolium. We also investigated the in vitro immune-enhancing activity of LLP-1A in macrophages and the underlying molecular mechanism. Results showed that LLP-1A was mainly composed of mannose and glucose at a molar ratio of 1.77:1, and its molecular weight was approximately 78.61kDa. The Fourier transform-infrared spectra of LLP-1A also revealed typical polysaccharide characteristics: the presence of uronic acid, pyranose rings and ß-glycosidic bonds. With regard to its effects on macrophages, LLP-1A enhanced phagocytic activity and induced the NO production in a dose-dependent manner. Further, it induced expression of the cytokines interleukin-6, monocyte chemotactic protein 1, tumor necrosis factor-α and interleukin-1ß. With regard to the molecular mechanism, LLP-1A increased protein expression of Toll-like receptor 4 and phosphorylation of the inhibitor of nuclear factor kappa-B kinase, inhibitor of NF-κB, and nuclear factor-kappa B in RAW 264.7 cells. Therefore, the data suggest that LLP-1A significantly upregulated the expression of immune reactive cytokines in RAW 264.7 macrophages through the TLR4-mediated NF-κB signal pathway. Thus, LLP-1A may have immunomodulatory functions that may prove beneficial for the treatment of immune-related diseases.

In vitro characterization of a (E)-ß-farnesene synthase from Matricaria recutita L. and its up-regulation by methyl jasmonate.

(E)-ß-farnesene is a sesquiterpene semiochemical that is used extensively by both plants and animals for communication. This acyclic olefin is found in the essential oil of chamomile (Matricaria recutita) and was demonstrated that it could attract natural enemies to reduce cabbage aphids in the Chinese cabbage fields. However, little is known regarding the sequence and function of (E)-ß-farnesene synthase in M. recutita. In this study, we reported a new full-length cDNA encoding (E)-ß-farnesene synthase from M. recutita (Mr-ßFS). The cDNA of Mr-ßFS consisted of 2010bp including 1725bp of coding sequence encoding a protein of 574 amino acids with a molecular weight of 67kDa. The deduced amino acid sequence exhibits a considerably higher homology to ßFS from Artemisia annua (about 92% identity) than to ßFSs from other plants (about 20-40% identity). The recombinant enzyme, produced in Escherichia coli, catalyzed the formation of a single product, (E)-ß-farnesene, from farnesyl diphosphate. Real-time quantitative PCR (qRT-PCR) analysis showed that Mr-ßFS expression was highest in leaves and lowest in disk florets. The treatment of M. recutita with methyl jasmonate (MeJA) significantly enhanced the transcriptional level of ßFS gene and the content of (E)-ß-farnesene in M. recutita. The transcriptional level of ßFS gene was approximately 11.5-fold higher than the control sample and the (E)-ß-farnesene emission level ranged from approximately from 0.082 to 0.695µg/g after 24h induction. Our results laid a solid foundation for later improving crop aphid resistance by transgenic technology and provided an important basic data for the regulation of valuable products from M. recutita.