Identification and validation of tumor-specific T cell receptors from tumor infiltrating lymphocytes using tumor organoid co-cultures.

T cell receptor-engineered T cells (TCR-Ts) therapy is promising for cancer immunotherapy. Most studies have focused on identifying tumor-specific T cell receptors (TCRs) through predicted tumor neoantigens. However, current algorithms for predicting tumor neoantigens are unreliable and many neoantigens are derived from non-coding regions. Thus, the technological platform for identifying tumor-specific TCRs using natural antigens expressed on tumor cells is urgently needed. In this study, tumor organoids-enriched tumor infiltrating lymphocytes (oeT) were obtained by repeatedly stimulation of autologous patient-derived organoids (PDO) in vitro. The oeT cells specifically responded to autologous tumor PDO by detecting CD137 expression and the secretion of IFN-γ using enzyme-linked immunospot assay. The measurement of oeT cell-mediated killing of three-dimensional organoids was conducted using a caspase3/7 flow cytometry assay kit. Subsequently, tumor-specific T cells were isolated based on CD137 expression and their TCRs were identified through single-cell RT-PCR analysis. The specificity cytotoxicity of TCRs were confirmed by transferring to primary peripheral blood T cells. The co-culture system proved highly effective in generating CD8+ tumor-specific oeT cells. These oeT cells effectively induced IFN-γ secretion and exhibited specificity in killing autologous tumor organoids, while not eliciting a cytotoxic response against normal organoids. The analysis conducted by TCRs revealed a significant expansion of T cells within a specific subset of TCRs. Subsequently, the TCRs were cloned and transferred to peripheral blood T cells generation engineered TCR-Ts, which adequately recognized and killed tumor cell in a patient-specific manner. The co-culture system provided an approach to generate tumor-specific TCRs from tumor-infiltrating lymphocytes of patients with colorectal cancer, and tumor-specific TCRs can potentially be used for personalized TCR-T therapy.

Functional Textile Materials for Blocking COVID-19 Transmission.

The outbreak of COVID-19 provided a warning sign for society worldwide: that is, we urgently need to explore effective strategies for combating unpredictable viral pandemics. Protective textiles such as surgery masks have played an important role in the mitigation of the COVID-19 pandemic, while revealing serious challenges in terms of supply, cross-infection risk, and environmental pollution. In this context, textiles with an antivirus functionality have attracted increasing attention, and many innovative proposals with exciting commercial possibilities have been reported over the past three years. In this review, we illustrate the progress of textile filtration for pandemics and summarize the recent development of antiviral textiles for personal protective purposes by cataloging them into three classes: metal-based, carbon-based, and polymer-based materials. We focused on the preparation routes of emerging antiviral textiles, providing a forward-looking perspective on their opportunities and challenges, to evaluate their efficacy, scale up their manufacturing processes, and expand their high-volume applications. Based on this review, we conclude that ideal antiviral textiles are characterized by a high filtration efficiency, reliable antiviral effect, long storage life, and recyclability. The expected manufacturing processes should be economically feasible, scalable, and quickly responsive.

Immune landscape of viral cancers: Insights from single-cell sequencing.

Viral infections trigger a wide range of immune responses thought to drive tumorigenesis and malignant progression. Dissecting virus-induced changes in the tumor immune microenvironment (TIME) is therefore crucial to identify key leukocyte populations that may represent novel targets for cancer therapy. Single-cell sequencing approaches have now been widely applied to the analysis of various tumors, thus enabling multiomics characterization of the highly heterogeneous TIME that bulk-sequencing cannot fully elucidate. In this review, we summarized key recent findings from sequencing studies of the immune infiltrate and antitumor response in virus-associated cancers at single cell resolution. Additionally, we also reviewed recent developments in immunotherapy for virus-associated cancers. We anticipate that the strategic use of single-cell sequencing will advance our understanding of the TIME of viral cancers, leading to the development of more potent novel treatments.

Progressive Design of a Ranatuerin-2 Peptide from Amolops wuyiensis: Enhancement of Bioactivity and In Vivo Efficacy.

Antimicrobial peptides (AMPs) that exert multiple functions are considered promising candidates to combat the bacterial drug resistance crisis. Nowadays, targeted peptide modification has been widely recognised to improve biological activity and make up for deficiencies in clinical applications such as toxicity. In this study, a helix-loop peptide was isolated and identified from the skin secretion of the Wuyi torrent frog Amolops wuyiensis, namely, ranatuerin-2-AW (R2AW) (GFMDTAKNVAKNVAATLLDKLKCKITGGC). Target modifications were made to R2AW to study the structure-activity relationships and to optimise its bioactivities. Five analogues were progressively designed via residue substitution and truncation and the antibacterial and anticancer activities were evaluated. We found that the serine-substitution and cyclic-domain-deletion products showed similar antibacterial activity to the natural peptide R2AW, implying that the disulphide bridge and Rana box were dispensable for the antibacterial activity of ranatuerin-2 peptides. Notably, the cationicity- and hydrophobicity-enhanced variant, [Lys4,19, Leu20]R2AW(1-22)-NH2, exhibited significantly optimised antibacterial and anticancer activities. Additionally, it killed bacteria by membrane disruption at a highly efficient rate. Moreover, [Lys4,19, Leu20]R2AW(1-22)-NH2 exerted potential in vivo efficacy in a methicillin-resistant Staphylococcus aureus (MRSA)-infected waxworm model. Overall, this study demonstrated some rational design ideas for optimising the dual antibacterial and anticancer activities of ranatuerin-2 peptides and it proposes [Lys4,19, Leu20]R2AW(1-22)-NH2 as an appealing candidate for therapeutic development.

Interleukin-33 as an early predictor of cetuximab treatment efficacy in patients with colorectal cancer.

BACKGROUND: Cetuximab is used for colorectal cancer (CRC) treatment. However, the early biomarker of treatment efficacy of cetuximab has not been identified. METHODS: After 1 year of cetuximab treatment, patients were divided into an effective group and an ineffective group. The interleukin-33 (IL-33) level and the distribution of lymphatic cells in patients were investigated by analyzing the peripheral blood mononuclear cells via flow cytometry analysis and ELISA. The correlation between IL-33 immunomodulatory effect and cetuximab treatment efficacy was determined through experiments in vivo and in vitro. RESULTS: The IL-33 level in the peripheral blood was increased at 4 weeks after cetuximab administration of effective group, meanwhile, the osteopontin (OPN) was reduced. Whereas neither IL-33 level nor OPN level of ineffective patients changed. In the effective group, the number of natural killer (NK) and CD8+ T cells were increased. Moreover, CD137 and CD107a expression on NK cells were higher in the effective group compared to the ineffective group. In vitro cetuximab treatment also increased the number of NK and CD8+ T cells as well as CD137 and CD107a expression upon IL-33 stimulation. Moreover, the secretion of OPN was inhibited by IL-33 administration in cetuximab-treated PBMCs from the effective group patients. IL-33 upregulated the cytotoxicity of NK cells and inhibited tumor cells growth in the effective cetuximab treatment mice. CONCLUSION: Effective cetuximab treatment induced a change of IL-33 and OPN at the early stage and triggered the NK cells antitumor activity. Consequently, significantly increased IL-33 level and decreased OPN level in the peripheral blood at the early treatment are proposed as potential predictors of cetuximab treatment efficacy.

Evolution of akirin family in gene and genome levels and coexpressed patterns among family members and rel gene in croaker.

Akirins, which are highly conserved nuclear proteins, are present throughout the metazoan and regulate innate immunity, embryogenesis, myogenesis, and carcinogenesis. This study reports all akirin genes from miiuy croaker and analyzes comprehensively the akirin gene family combined with akirin genes from other species. A second nuclear localization signal (NLS) is observed in akirin2 homologues, which is not in akirin1 homologues in all teleosts and most other vertebrates. Thus, we deduced that the loss of second NLS in akirin1 homologues in teleosts likely occurred in an ancestor to all Osteichthyes after splitting with cartilaginous fish. Significantly, the akirin2(2) gene included six exons interrupted by five introns in the miiuy croaker, which may be caused by the intron insertion event as a novel evidence for the variation of akirin gene structure in some species. In addition, comparison of the genomic neighborhood genes of akirin1, akirin2(1), and akirin2(2) demonstrates a strong level of conserved synteny across the teleost classes, which further proved the deduction of Macqueen and Johnston 2009 that the produce of akirin paralogues can be attributed to whole-genome duplications and the loss of some akirin paralogues after genome duplications. Furthermore, akirin gene family members and relish gene are ubiquitously expressed across all tissues, and their expression levels are increased in three immune tissues after infection with Vibrio anguillarum. Combined with the expression patterns of LEAP-1 and LEAP-2 from miiuy croaker, an intricate network of co-regulation among family members is established. Thus, it is further proved that akirins acted in concert with the relish protein to induce the expression of a subset of downstream pathway elements in the NF-kB dependent signaling pathway.

Isolation, identification and antioxidant activity of bound phenolic compounds present in rice bran.

The bound phenolic compounds in rice bran were released and extracted with ethyl acetate based on alkaline digestion. An investigation of the chemical constituents of EtOAc extract has led to the isolation of a new compound, para-hydroxy methyl benzoate glucoside (8), together with nine known compounds, cycloeucalenol cis-ferulate (1), cycloeucalenol trans-ferulate (2), trans-ferulic acid (3), trans-ferulic acid methyl ester (4), cis-ferulic acid (5), cis-ferulic acid methyl ester (6), methyl caffeate (7), vanillic aldehyde (9) and para-hydroxy benzaldehyde (10). The structures of these compounds were determined using a combination of spectroscopic methods and chemical analysis. Among the compounds isolated, compound 3, 5 and 7 exhibited strong DPPH and ABTS(+) radical scavenging activities, followed by compounds 4 and 6. Compound 1 and 2 showed potent DPPH and ABTS(+) radical scavenging activities, compound 8 displayed moderate antioxidant activity against ABTS(+) radical, whereas compound 9 and 10 showed weak antioxidant activity.

New phenolic compounds from Coreopsis tinctoria Nutt. and their antioxidant and angiotensin i-converting enzyme inhibitory activities.

Three new phenolic compounds, coretinphenol (1), coretincone (2), and coretinphencone (3), were isolated from the buds of Coreopsis tinctoria Nutt., together with nine known compounds, including butein (4), okanin (5), isoliquiritigenin (6), maritimetin (7), taxifolin (8), isookanin (9), marein (10), sachalinoside B (11), and 2-phenylethyl-ß-d-glucoside (12). The chemical structures of these compounds were elucidated by extensive spectroscopic analysis and on the basis of their chemical reactivity. This work represents the first recorded example of the isolation of compounds 1­3, 6, 7, 9, 11, and 12 from C. tinctoria. Compounds 5­9 showed strong diphenyl(2,4,6-trinitrophenyl)iminoazanium (DPPH) radical-scavenging activity, with IC50 values of 3.35 ± 0.45, 9.6 ± 2.32, 4.12 ± 0.21, 6.2 ± 0.43, and 7.9 ± 0.53 µM, respectively. Compounds 2 and 8 exhibited angiotensin I-converting enzyme inhibitory activity, with IC50 values of 228 ± 4.47 and 145.67 ± 3.45 µM, respectively. The activities of phenolic compounds isolated from C. tinctoria support the medicinal use of this plant in the prevention of cardiovascular diseases.

Characterization, evolution and functional analysis of the liver-expressed antimicrobial peptide 2 (LEAP-2) gene in miiuy croaker.

As an evolutionarily older defense strategy, the innate immune is the dominant immune system and provides a first line of antimicrobial host defense in teleost. Liver-expressed antimicrobial peptide-2 (LEAP-2) is a critical molecule of the innate immune system and plays a very important role in resistance against bacterial infections. We reported comprehensive analysis and characterization of LEAP-2 gene from miiuy croaker (Miichthys miiuy) in here. The complete cDNA of miiuy croaker LEAP-2 consists 2360 bp, including a 5' terminal untranslated region (UTR) of 170 bp, an open reading frame (ORF) of 312 bp, and a 3'-UTR of 1878 bp. Interestingly, two polyadenylation signals (AATTAAA) which may involve the stability, translation efficiency, or localization of an mRNA in a tissue were found in 3'-UTR. Genomic DNA of miiuy croaker LEAP-2 includes three exons and two introns, which is similar to LEAP-2 genes in other mammals and fish. The deduced 103 amino acids consist of signal peptide, prodomain and mature peptide. Four highly conserved cysteine residues involved two disulfide bridges in mature peptide. Real-time PCR results showed that LEAP-2 was ubiquitously expressed in all tissues and the expression level was highest in liver. Significantly, the expression levels were increased after infection with Vibrio anguillarum in liver and spleen. The antimicrobial activity analysis result of LEAP-2 in vitro indicated that LEAP-2 of miiuy croaker was effective in controlling Aeromonas hydrophila. In addition, we performed evolutionary analysis in order to estimate the selective constraints on the LEAP-2 gene. The result indicated that no positive selection exists in LEAP-2 gene sequences, which may be on account of irreplaceable function constrains. Meanwhile, we compared the structure of LEAP-2 with that of another Liver-expressed antimicrobial peptide (LEAP-1, also named HAMP), and found the LEAP-2 from miiuy croaker comprises of α-helix, ß-sheet, and ß-turn while the LEAP-1 of miiuy croaker only contains ß-sheet.

Two new noroleanane-type triterpene saponins from the methanol extract of Salicornia herbacea.

Two new noroleanane-type triterpene saponins, Salbige A (1) and Salbige B (2), have been isolated from the aerial parts of Salicornia herbacea together with five other known compounds, including echinocystic acid (3), gypsogenin (4), pheophorbide a (5), (13(2)S)-hydroxy-pheophorbide a (6) and (13(2)S)-hydro-pheophorbide-lactone a (7). The chemical structures of these compounds were elucidated by extensive spectroscopic analysis and on the basis of their chemical reactivity. This work represents the first recorded example of the isolation of these compounds from S. herbacea. Compounds 1 and 2 exhibited potent antiproliferative activities and high levels of selectivity towards A549 cancer cells, with IC50 values of 52.35 and 79.39 µM, respectively, whereas compound 5 showed high levels of inhibitory activity against A549 and HepG2 cancer cells with IC50 values of 6.15 and 17.56 µM, respectively. None of these compounds exhibited antioxidant activities except for compound 7, which showed weak antioxidant activity.

Pentadecyl ferulate, a potent antioxidant and antiproliferative agent from the halophyte Salicornia herbacea.

An investigation of the chemical constituents of Salicornia herbacea has led to the isolation of one new natural product, pentadecyl ferulate (6), together with 11 known compounds, including phytol (1), stearolic acid (2), γ-linolenic acid (3), (3Z,6Z,9Z)-tricosa-3,6,9-triene (4), linoleic acid (5), stigmasterol (7), ergosterol (8), dioctyl phthalate (9), dibutyl phthalate (10), vanillic aldehyde (11), and scopoletin (12). The chemical structures of these materials were elucidated mainly by spectroscopic analysis. This work represents the first recorded example of the isolation of compounds 1, 2, 3, 4, 9, 10, and 11 from S. herbacea. The antioxidant experiments revealed that compound 6 possessed strong hydroxy radical and superoxide anion scavenging activities and was the principle antioxidant ingredient in the ethyl acetate extract. The antiproliferative results exhibited that compound 1 selectively inhibited HepG2 cells, whereas compounds 3 and 6 showed potent antiproliferative activities against HepG2 and A549 cells.

Long-term follow up of very low-dose LINAC based stereotactic radiotherapy in temporal lobe epilepsy.

OBJECTIVE: Several prospective multicenter studies have demonstrated high safety efficacy for mesial temporal lobe epilepsy (TLE) with Gamma Knife radiosurgery using 24 Gy at the margin. In this paper we reported the long-term outcome of 7 patients with TLE treated with very low-dose linear accelerator (LINAC) based fractionated stereotactic radiotherapy (FSRT). We also discussed the feasibility of LINAC for TLE. METHODS: To record the treatment process, outcomes and side effects for 7 patients with TLE treated with low-dose LINAC, whose marginal dose of all cases was 12 Gy at the 85% isodose line, and post-FSRT seizure frequencies were compared with those of patients pre-FSRT. One special case (case 6) was reported in detail. RESULTS: Reduction of seizure frequency post-FSRT was 50% in 2 cases, 30% in 1 case, and 0% in 2 cases, and seizure frequency increased more than 100% in 2 cases. No patient was seizure free at the last follow up. 2 cases presented transitory complications and 2 cases showed an obvious drop in IQ, memory decline and permanent neurologic complications, including partial aphasia and mild hemiplegia in 1 case, progressive ataxia and cognition decline in another case. CONCLUSION: Outcome of TLE treated by very low-dose LINAC based FSRT was poor because of mild reduction in seizure attack frequency and serious complications.

[Therapeutic effect of three dimensional conformal radiotherapy in patients aged 65 years and older with non-small cell lung cancer].

BACKGROUND: Three dimensional conformal radiotherapy is one of the important treatment methods for patients aged 65 and older with inoperable non-small cell lung cancer (NSCLC). The aim of this study is to evaluate the efficacy of three dimensional conformal radiotherapy in elderly patients with inoperable NSCLC. METHODS: Thirty-one patients aged 65 and older with stage II and III NSCLC were treated by three dimensional conformal radiotherapy. Clinical effects, survival rates and pulmonary toxicity were analyzed. RESULTS: The overall response rate (CR+PR) was 87.1% (27/31). The 1, 2 and 3-year survival rates were 83.9% (26/31), 51.6% (16/31), and 25.8% (8/31) respectively. The relation of treatment efficacy and tumor size was observed. Acute radiation pneumonitis occurred in 3.2% (1/31) of the patients. Radiation pulmonary fibrosis occurred in 19.4% (6/31) of the patients. CONCLUSIONS: Three dimensional conformal radiotherapy is a safe and effective treatment for elderly patients with inoperable NSCLC and has a high survival rate.