Compound heterozygous WNT10A missense variations exacerbated the tooth agenesis caused by hypohidrotic ectodermal dysplasia.

BACKGROUND: The aim of this study was to analyse the differences in the phenotypes of missing teeth between a pair of brothers with hypohidrotic ectodermal dysplasia (HED) and to investigate the underlying mechanism by comparing the mutated gene loci between the brothers with whole-exome sequencing. METHODS: The clinical data of the patients and their mother were collected, and genomic DNA was extracted from peripheral blood samples. By Whole-exome sequencing filtered for a minor allele frequency (MAF) ≤0.05 non-synonymous single-nucleotide variations and insertions/deletions variations in genes previously associated with tooth agenesis, and variations considered as potentially pathogenic were assessed by SIFT, Polyphen-2, CADD and ACMG. Sanger sequencing was performed to detect gene variations. The secondary and tertiary structures of the mutated proteins were predicted by PsiPred 4.0 and AlphaFold 2. RESULTS: Both brothers were clinically diagnosed with HED, but the younger brother had more teeth than the elder brother. An EDA variation (c.878 T > G) was identified in both brothers. Additionally, compound heterozygous variations of WNT10A (c.511C > T and c.637G > A) were identified in the elder brother. Digenic variations in EDA (c.878 T > G) and WNT10A (c.511C > T and c.637G > A) in the same patient have not been reported previously. The secondary structure of the variant WNT10A protein showed changes in the number and position of α-helices and ß-folds compared to the wild-type protein. The tertiary structure of the WNT10A variant and molecular simulation docking showed that the site and direction where WNT10A binds to FZD5 was changed. CONCLUSIONS: Compound heterozygous WNT10A missense variations may exacerbate the number of missing teeth in HED caused by EDA variation.

Dual-emissive carbon dots for dual-channel ratiometric fluorometric determination of pH and mercury ion and intracellular imaging.

Dual-emissive carbon dots (CDs) were fabricated for dual-channel ratiometric fluorometric determination of pH and mercury ion (Hg2+) and intracellular imaging. Dual-emissive CDs were synthesized by one-pot solvothermal treatment of cabbage. The CDs exhibited two distinctive fluorescence emissions at 500 and 678 nm under single excitation at 410 nm. The green emission (500 nm) had reversible linear response to pH (7.0-12.0) due to deprotonation and protonation of surface functional groups and their non-covalent interactions. On the other hand, the red emission (678 nm) had efficient and selective fluorescence response to Hg2+ by formation of non-emission complex between CDs and Hg2+. The limit of detection (LOD) and limit of quantification (LOQ) for Hg2+ were 6.25 and 20.63 nM, respectively. The CDs have been successfully applied for label-free ratiometric fluorometric determination of pH and Hg2+ in fish and human serum samples with good recoveries (92.0-108.3%). In addition, the CDs had excellent photostability, low cytotoxicity, and good biocompatibility for intracellular imaging. All in all, the system was multi-functional in determination, high in sensitivity, and excellent in selectivity, which demonstrated wide and promising applicability for biosensing and bioimaging in the future. Graphical abstract Schematic presentation of dual-emission carbon dots (CDs) synthesized by solvothermal treatment of cabbage for dual-channel determination of pH and Hg2+.

Effects of different anti-caries agents on microhardness and superficial microstructure of irradiated permanent dentin: an in vitro study.

BACKGROUND: To compare different anti-caries agents on microhardness and micromorphology of irradiated permanent dentin in vitro, and try to find the most effective agent to prevent radiation-dentin-destruction. METHODS: A total of 120 dentin samples were prepared from 60 human teeth and randomly divided into 8 groups (n = 15), [ (1)] blank control [2]; irradiation control [3]; irradiation+ fluoride [4]; irradiation+ casein phosphate polypeptide-amorphous calcium phosphate (CPP-ACP) [5]; irradiation+ CPP-ACP+ fluoride [6]; irradiation+ infiltration resin [7]; irradiation+ infiltration resin+ fluoride [8]; irradiation+ infiltration resin+ CPP-ACP. Seven samples of each groups were chosen randomly for microhardness test and eight for scanning electron microscope observation. RESULTS: A decrease of microhardness (P < 0.05) and an obvious morphological change were presented on dentin surface after radiotherapy. After applications of anti-caries agents, the morphological destructions were effectively restored. The infiltration resin plus fluoride group (56.00 ± 4.02 Kg/mm2), infiltration resin plus CPP-ACP group (56.05 ± 3.69 Kg/mm2), infiltration resin group (54.70 ± 4.42Kg/mm2) and CPP-ACP plus fluoride group (53.84 ± 6.23Kg/mm2) had the highest dentin microhardness value after radiotherapy, and no statistically significant difference were found between them. CONCLUSIONS: Infiltration resin, CPP-ACP, fluoride and their pairwise combination can effectively prevent radiation-dentin-destruction. Among them, infiltration resin with CPP-ACP, infiltration resin with fluoride, CPP-ACP with fluoride, and infiltration resin have the most protective effects on irradiation-dentin-destructions.

Engineering chimeric antigen receptor-T cells for cancer treatment.

Intratumor heterogeneity of tumor clones and an immunosuppressive microenvironment in cancer ecosystems contribute to inherent difficulties for tumor treatment. Recently, chimeric antigen receptor (CAR) T-cell therapy has been successfully applied in the treatment of B-cell malignancies, underscoring its great potential in antitumor therapy. However, functional challenges of CAR-T cell therapy, especially in solid tumors, remain. Here, we describe cancer-immunity phenotypes from a clonal-stromal-immune perspective and elucidate mechanisms of T-cell exhaustion that contribute to tumor immune evasion. Then we assess the functional challenges of CAR-T cell therapy, including cell trafficking and infiltration, targeted-recognition and killing of tumor cells, T-cell proliferation and persistence, immunosuppressive microenvironment and self-control regulation. Finally, we delineate tumor precision informatics and advancements in engineered CAR-T cells to counteract inherent challenges of the CAR-T cell therapy, either alone or in combination with traditional therapeutics, and highlight the therapeutic potential of this approach in future tumor precision treatment.

Conservation analysis and pathogenicity prediction of mutant genes of ectodysplasin a.

BACKGROUND: Hypohidrotic ectodermal dysplasia (HED) is a common recessive X-linked hereditary disease that affects the development of ectoderm. Gene mutations of ectodysplasin A (EDA) play key roles in process of this disease. In our preliminary study, three unknown mutation sites (c.878 T > G, c.663-697del and c.587-615del) were detected from the pedigrees of HED. METHODS: Conservation analysis of the related homologous proteins in 3 unknown EDA gene mutation sites was conducted using the University of California Santa Cruz (UCSC) Genome Browser database. SIFT and PolyPhen-2, the online gene function prediction software, were utilized to predict the pathogenicity of point mutation of c.878 T > G. RESULTS: All three unknown mutation sites were located in the highly-conserved region of EDA and possessed strong amino acid conservation among different species. In addition, the results of the pathogenicity prediction of point mutation of c.878 T > G by SIFT (P = 0.00) and PolyPhen-2 (S = 0.997) demonstrated that the mutation site had considerable pathogenicity theoretically. CONCLUSIONS: The EDA mutations of c.878 T > G, c.663-697del and c.587-615del may be responsible for the pathogenesis of HED in their pedigrees.

Meisoindigo, but not its core chemical structure indirubin, inhibits zebrafish interstitial leukocyte chemotactic migration.

CONTEXT: Inflammatory disease is a big threat to human health. Leukocyte chemotactic migration is required for efficient inflammatory response. Inhibition of leukocyte chemotactic migration to the inflammatory site has been shown to provide therapeutic targets for treating inflammatory diseases. OBJECTIVE: Our study was designed to discover effective and safe compounds that can inhibit leukocyte chemotactic migration, thus providing possible novel therapeutic strategy for treating inflammatory diseases. MATERIALS AND METHODS: In this study, we used transgenic zebrafish model (Tg:zlyz-EGFP line) to visualize the process of leukocyte chemotactic migration. Then, we used this model to screen the hit compound and evaluate its biological activity on leukocyte chemotactic migration. Furthermore, western blot analysis was performed to evaluate the effect of the hit compound on the AKT or ERK-mediated pathway, which plays an important role in leukocyte chemotactic migration. RESULTS: In this study, using zebrafish-based chemical screening, we identified that the hit compound meisoindigo (25 µM, 50 µM, 75 µM) can significantly inhibit zebrafish leukocyte chemotactic migration in a dose-dependent manner (p = 0.01, p = 0.0006, p < 0.0001). Also, we found that meisoindigo did not affect the process of leukocyte reverse migration (p = 0.43). Furthermore, our results unexpectedly showed that indirubin, the core structure of meisoindigo, had no significant effect on zebrafish leukocyte chemotactic migration (p = 0.6001). Additionally, our results revealed that meisoindigo exerts no effect on the Akt or Erk-mediated signalling pathway. DISCUSSION AND CONCLUSION: Our results suggest that meisoindigo, but not indirubin, is effective for inhibiting leukocyte chemotactic migration, thus providing a potential therapeutic agent for treating inflammatory diseases.

Bmp2 deletion causes an amelogenesis imperfecta phenotype via regulating enamel gene expression.

Although Bmp2 is essential for tooth formation, the role of Bmp2 during enamel formation remains unknown in vivo. In this study, the role of Bmp2 in regulation of enamel formation was investigated by the Bmp2 conditional knock out (Bmp2 cKO) mice. Teeth of Bmp2 cKO mice displayed severe and profound phenotypes with asymmetric and misshaped incisors as well as abrasion of incisors and molars. Scanning electron microscopy analysis showed that the enamel layer was hypoplastic and enamel lacked a typical prismatic pattern. Teeth from null mice were much more brittle as tested by shear and compressive moduli. Expression of enamel matrix protein genes, amelogenin, enamelin, and enamel-processing proteases, Mmp-20 and Klk4 was reduced in the Bmp2 cKO teeth as reflected in a reduced enamel formation. Exogenous Bmp2 up-regulated those gene expressions in mouse enamel organ epithelial cells. This result for the first time indicates Bmp2 signaling is essential for proper enamel development and mineralization in vivo.

Association Between TP53 Gene Codon 72 Polymorphism and Acute Myeloid Leukemia Susceptibility: Evidence Based on a Meta-Analysis.

BACKGROUND: Many studies have reported that the p53 codon 72 polymorphism is associated with acute myeloid leukemia (AML) susceptibility; however, the conclusions are inconsistent. Therefore, we performed this meta-analysis to obtain a more precise result. MATERIAL AND METHODS: We searched PubMed to identify relevant studies, and 6 published case-control studies were retrieved, including 924 AML patients and 3832 controls. Odds ratio (OR) with corresponding 95% confidence interval (95%CI) was applied to assess the association between p53 codon 72 polymorphism and AML susceptibility. The meta-analysis was performed with Comprehensive Meta-Analysis software, version 2.2. RESULTS: Overall, no significant association between p53 codon 72 polymorphism and AML susceptibility was found in this meta-analysis (Pro vs. Arg: OR=0.94, 95%CI=0.81-1.10; Pro/Pro vs. Arg/Arg: OR=0.93, 95%CI=0.71-1.22; Arg/Pro vs. Arg/Arg: OR=0.79, 95%CI=0.55-1.13; (Pro/Pro + Arg/Pro) vs. Arg/Arg: OR=0.84, 95%CI=0.62-1.13; Pro/Pro vs. (Arg/Arg + Arg/Pro): OR=1.06, 95%CI=0.83-1.35). Similar results were also found in stratified analysis according to ethnicity and source of controls. CONCLUSIONS: Our meta-analysis demonstrates that p53 codon 72 polymorphism may not be a risk factor for AML, which should be verified in future studies.

The oligodontia phenotype in a X-linked hypohidrotic ectodermal dysplasia patient with a novel EVC2 variant.

Objectives: To analyse the pathogenic genes in a patient with hypohidrotic ectodermal dysplasia (HED) and explore the relationship between pathogenic genes and the oligodontia phenotype. Methods: Clinical data and peripheral blood were collected from a patient with HED. Pathogenic genes were analysed by whole-exon sequencing (WES) and verified by Singer sequencing. The secondary and tertiary structures of the variant proteins were predicted to analyse their toxicity. Results: The patient exhibited a severe oligodontia phenotype, wherein only two deciduous canines were left in the upper jaw. WES revealed a hemizygous EDA variant c.466C > T p.(Arg156Cys) and a novel heterozygous EVC2 variant c.1772T > C p.(Leu591Ser). Prediction of the secondary and tertiary structures of the EDA variant p.(Arg156Cys) and EVC2 variant p.(Leu591Ser) indicated impaired function of both molecules. Conclusion: The patient demonstrated a more severe oligodontia phenotype when compared with the other patients caused by the EDA variant c.466C > T. Since Evc2 is a positive regulator of the Sonic Hedgehog (Shh) signal pathway, we speculated that the EVC2 variant p.(Leu591Ser) may play a synergistic role in the oligodontia phenotype of HED, thereby exacerbating the oligodontia phenotype. Knowledge of oligodontia caused by multiple gene variants is of great significance for understanding individual differences in oligodontia phenotypes.

Boosting electrocatalytic performance via electronic structure regulation for acidic oxygen evolution.

High-purity hydrogen produced by water electrolysis has become a sustainable energy carrier. Due to the corrosive environments and strong oxidizing working conditions, the main challenge faced by acidic water oxidation is the decrease in the activity and stability of anodic electrocatalysts. To address this issue, efficient strategies have been developed to design electrocatalysts toward acidic OER with excellent intrinsic performance. Electronic structure modification achieved through defect engineering, doping, alloying, atomic arrangement, surface reconstruction, and constructing metal-support interactions provides an effective means to boost OER. Based on introducing OER mechanism commonly present in acidic environments, this review comprehensively summarizes the effective strategies for regulating the electronic structure to boost the activity and stability of catalytic materials. Finally, several promising research directions are discussed to inspire the design and synthesis of high-performance acidic OER electrocatalysts.

Susceptibility artifacts induced by crowns of different materials with prepared teeth and titanium implants in magnetic resonance imaging.

This study aimed to investigate the artifacts induced by crowns composed of different materials with prepared teeth and titanium implants. Resin, metal-ceramic, ceramic and zirconia crowns were fabricated and placed onto the prepared teeth on a human cadaver head or titanium implants with prosthesis abutments on a dry human mandible. The samples were scanned on a 1.5 T MRI apparatus, and artifact areas were defined as the signal intensity and signal loss adjacent to the prosthesis and measured by a threshold tool with ImageJ2x. Data were analyzed using SPSS 22.0. Resin, ceramic, zirconia, and precious metal-ceramic crowns barely produced artifacts on the cadaver skull (p > 0.999). By contrast, pure Ti and nonprecious metal-ceramic crowns created significant artifacts (p < 0.001). The average artifacts reduction of double Au-Pt and Ag-Pd metal-ceramic crowns combined with titanium implants and abutments was 79.49 mm2 (p < 0.001) and 74.17 mm2 (p < 0.001) respectively, while artifact areas were increased in double Co-Cr and Ni-Cr metal-ceramic crowns by 150.10 mm2 (p < 0.001) and 175.50 mm2 (p < 0.001) respectively. Zirconia, ceramic and precious metal-ceramic crowns induce less MRI artifacts after tooth preparation while precious metal-ceramic crowns alleviate artifacts in combination with titanium implants.

[The Levels of Inflammatory Regulatory Factors in Acute Leukemia Complicated by Infection and Its Clinical Significance].

OBJECTIVE: To explore the expression levels and clinical significance of helper T cell 1/helper T cell 2 (Th1/Th2) cytokine and interleukin-6 (IL-6) in patients with acute leukemia (AL) complicated by infection. METHODS: 68 patients with AL complicated by infection admitted to Wuhan Fifth Hospital from May 2017 to January 2020 were enrolled as study group, 50 AL patients without infection were enrolled as AL group, and 30 healthy volunteers checked in physical examination center were enrolled as healthy control group. The levels of serum tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10), and peripheral blood Th1/Th2 cells subsets were measured and compared among the three groups. The serum IL-6, IL-10, TNF-α and Th1/Th2 were compared between the patients with mild to moderate infection (n=52) and septic shock (n=16). The relationship between IL-6, IL-10, TNF-α, Th1/Th2 and AL infection was analyzed. RESULTS: The levels of IL-6, IL-10 , TNF-α, and the proportion of Th2 of the patients in study group and AL group were significantly higher than those in healthy control group (P<0.001), while the proportion of Th1 and Th1/Th2 were significantly lower than those in healthy control group (P<0.001). The levels of IL-6, IL-10 and TNF-α, and the proportion of Th2 the patients in study group were significantly higher than those in AL group (P<0.001), while the proportion of Th1 and Th1/Th2 were significantly lower than those in AL group (P<0.001). The serum IL-6, IL-10 and TNF-α level of the patients in septic shock group were significantly higher than those in mild-to-moderate infection group (P<0.001), while Th1/Th2 was lower than those in mild-to-moderate infection group (P<0.001). The results of ROC curve analysis showed that the area under the ROC curve (AUC) values of IL-6, IL-10, TNF-α and Th1/Th2 alone for the diagnosis of septic shock were 0.779, 0.761, 0.724 and 0.718, which were lower than that their combination (0.910) (P<0.05). CONCLUSION: The levels of serum IL-6, IL-10 and TNF-α are high in patients with AL complicated infection and septic shock, while Th1/Th2 cell subsets is low. The combined detection of serum IL-6, IL-10, TNF-α and Th1/Th2 is a good diagnostic value for predicting the occurrence of severe septic shock.

Effects of different anti-caries procedures on microhardness and micromorphology of irradiated permanent enamel.

To explore the effects of infiltration resin, casein phosphate polypeptide-amorphous calcium phosphate (CPP-ACP) and fluoride on microhardness and micromorphology of irradiated enamel. Sixty human permanent teeth were mesiodistally sectioned, yielding 120 enamel samples, which randomly divided into 8 groups: G1: blank control; G2: irradiation control; G3: irradiation+fluoride; G4: irradiation+CPP-ACP; G5: irradiation+CPP-ACP+fluoride; G6: irradiation+infiltration resin; G7: irradiation+infiltration resin+fluoride; G8: irradiation+infiltration resin+CPP-ACP. A progressive improvement was observed on the superficial morphology of enamel treated with different anti-caries procedures. The order of microhardness values after irradiation from the highest to the lowest was as follows: G1>G8~G7>G6>G5>G4~G3>G2. CPP-ACP, infiltration resin and fluoride can effectively restore the direct destruction of enamel caused by irradiation and promote the occurrence of remineralization. Infiltration resin and its combined effects with fluoride or CPP-ACP have the most potential anti-caries agent to resist radiation-caries.

[Periodontal status in patients with oral submucous fibrosis].

OBJECTIVE: To investigate the periodontal status in patients with oral submucous fibrosis (OSF), and to provide reference for the treatment and prophylaxis in patients with OSF and betel chewers. METHODS: Fifty samples clinically and pathologically diagnosed as OSF patients were selected as the OSF group, another 50 age-matched healthy volunteers in the similar living condition were compared with the OSF patients and non-betel nut chewers were classified as the control group. The 5 periodontal clinical parameters were collected and recorded, including plaque index, periodontal probing depth, clinical attachment loss, gingival index, and tooth count of bleeding of probing. RESULTS: There was a significant difference in plaque index (PLI) between the OSF group (2.14+/-0.64) and the control group (1.7+/-0.89) (P<0.01). Periodontal probing depth (PD) was (1.98+/-0.70) mm in the control group, and (5.57+/-2.39) mm in the OSF group, with significant difference in PD (P<0.01). There was no significant difference in clinical attachment loss, gingival index, and tooth count of bleeding on probing between the 2 groups (P>0.05). CONCLUSION: OSF patients tend to accumulate plaque, and have deep periodontal pocket, periodontal inflammation or severe periodontal damage.

Early Caries Preventive Effects of Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP) Compared with Conventional Fluorides: A Meta-analysis.

PURPOSE: To assess the current evidence regarding the early caries preventive effects of CPP-ACP compared with fluorides. MATERIALS AND METHODS: An electronic search from PubMed, Cochrane Library, EMBASE, Web of Science and ScienceDirect database with a complementary gray literature search for randomised controlled human clinical trials were carried out. No language restrictions were applied. RESULTS: A total of 395 participants in 10 studies of the 600 selected studies were carefully reviewed and selected for inclusion in this meta-analysis. The pooling data of the remineralisation scores in vivo showed a weighted mean difference (WMD) in favor of CPP-ACP as compared to fluorides (WMD: -2.47; 95% confidence interval [CI]: [-3.62, -1.32]; p < 0.0001). The DMFS/dmfs (decayed, missing, filled surfaces) index and the Enamel Decalcification Index (EDI) did not differ significantly between CPP-ACP and fluorides. No serious side effects associated with CPP-ACP and fluoride agents were found. CONCLUSION: CPP-ACP may offer a safe and efficient alternative to fluorides with less mineral content loss (laser fluorescence values) and more remineralisation of early caries lesions. In view of the inherent limitations of the included researches, high-quality, well-designed randomised controlled trials are still needed. CPP-ACP has potential utility in promoting enamel remineralisation of early caries lesions compared with fluoride.

[Analysis of the risk factors of radiation-induced caries in patients with head and neck cancer].

OBJECTIVE: This study aimed to evaluate the risk factors of radiation-induced caries by using a multiple linear regression equation and to provide the basis for the effective prevention of radioactive caries. METHODS: A total of 166 patients with head and neck cancer who underwent radiotherapy were selected as subjects. The number of decayed, missing or filled surfaces were recorded. Questionnaire contents included age, sex, radiation dose, and radiotherapy techniques. Multiple stepwise regression analyses were performed to identify the risk factors of radiation-induced caries. RESULTS: Multiple stepwise regression analyses indicated that the main risk factors of radiation-induced caries were plaque index, radiotherapy techniques, time after radiotherapy, and radiotherapy dose. CONCLUSIONS: The awareness of dental care and caries treatment should be improved to reduce the occurrence of radiation-induced caries in patients with head and neck cancer. In addition, intensity modulated radiation therapy should be employed to decrease the radiation exposure dose received by teeth.

[Research Progress on the Role of Pyroptosis in the Pathogenesis of Myelodysplastic Syndrome -Review].

Pyroptosis is a novel type of programmed cell death, which is closely related with the pathogenesis of myelodysplastic syndromes (MDS). The recent studies showed that all of S100A9/TLR4, S100A9/CD33 and Nox/ROS signaling pathways can activate oxygen-sensitivity NLRP3 inflammasome and then induce the pyroptosis of hematopoeitic stem cells (HSC) / hematopeitic pregenitor cells (HPC), resulting in ineffective hematopoiesis in patients with MDS. Further studies on the role and molecular mechanism of pyroptosis in the pathogenesis of MDS will provide the potential opportunity for the diagnosis and treatment of MDS. Here, the recent advances in the role and mechnism of pyroptosis in the pathogenesis of MDS are reviewed.

[CRISPR/Cas9 System and Its Advances in Gene Therapy of Hematologic Diseases--Review].

In recent years, with the development of gene editing technology, the site-specific genome can be modified. The curability of genetic disease may be achieved by the use of gene editing techniques. As the simplicity, high specificity and economical efficiency, much attention has been paid to the CRISPR/Cas9 system, which was been widely used in research of molecular biology and other fields of life science. In this review, the mechanism for CR1SPR/Cas9 system and the progress of gene therapy, such as for hemophilia, betathalassaemia and chronic myeloid leukemia were summarized briefly.

High-throughput sequencing of the immune repertoire in oncology: Applications for clinical diagnosis, monitoring, and immunotherapies.

The diagnostic, monitoring and therapeutic options for cancers currently remain limited. These limitations represent a large threat to human health. Adaptive immunity, which is dependent on diverse repertoires of B cell receptors (BCRs) and T cell receptors (TCRs), plays a critical role in the anti-tumor immune response. Modulation and surveillance of adaptive immunity has become a powerful weapon to combat cancers. Recently, the high-throughput sequencing of immune repertoire (HTS-IR) technology, which provides a robust tool for deep sequencing repertoires of BCRs or TCRs, has been applied in the development of tumor biomarkers and immunotherapeutics for cancers. This review will first provide an overview of the advancement of HTS-IR technology at the population-cell and single-cell levels. It will then provide a current summary of the applications of HTS-IR technology in the diagnosis and monitoring of minimal residual disease (MRD), focusing on immune reconstitution after the treatment of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in B/T-cell malignancies, and the precise detection of tumor-infiltrating lymphocytes (TILs) in non-B/T-cell malignancies. Finally, current advances of HTS-IR technology in cancer immunotherapeutic applications, such as therapeutic antibodies, CAR-T cell based-adoptive immunotherapies, and neoantigen-specific TCR-T cell-based adoptive immunotherapies, will be introduced.

Antioxidant profiling of vine tea (Ampelopsis grossedentata): Off-line coupling heart-cutting HSCCC with HPLC-DAD-QTOF-MS/MS.

Vine tea with strong antioxidant activity is commonly consumed as healthy tea/beverage. However, detailed information about its antioxidants is incomplete. Here, off-line hyphenation of heart-cutting high-speed countercurrent chromatography (HSCCC) with high performance liquid chromatography-diode array detector-quadrupole time-of-flight tandem mass spectrometry (HPLC-DAD-QTOF-MS/MS) were described for systematic profiling antioxidants in vine tea. At first, antioxidants were rapidly screened by 1,1-diphenyl-2-picryl-hydrazyl radical-high performance liquid chromatography (DPPH-HPLC). Subsequently, stepwise HSCCC using petroleum ether-ethyl acetate-methanol-water (4:9:4:9, v/v/v/v) and (4:9:5:8, v/v/v/v) as solvent systems was optimized to fractionate and enrich antioxidants from ethyl acetate fraction of vine tea. Finally, heart-cutting mode was used to collect five interesting HSCCC fractions for HPLC-DAD-QTOF-MS/MS analysis. Desirable orthogonality between HSCCC and HPLC led to identification of fifteen antioxidant flavonoids, while four minor flavonoids were first reported in vine tea. Results showed that the developed system is efficient to comprehensively explore antioxidants from complex natural herbs.