Deep learning on tertiary lymphoid structures in hematoxylin-eosin predicts cancer prognosis and immunotherapy response.

Tertiary lymphoid structures (TLSs) have been associated with favorable immunotherapy responses and prognosis in various cancers. Despite their significance, their quantification using multiplex immunohistochemistry (mIHC) staining of T and B lymphocytes remains labor-intensive, limiting its clinical utility. To address this challenge, we curated a dataset from matched mIHC and H&E whole-slide images (WSIs) and developed a deep learning model for automated segmentation of TLSs. The model achieved Dice coefficients of 0.91 on the internal test set and 0.866 on the external validation set, along with intersection over union (IoU) scores of 0.819 and 0.787, respectively. The TLS ratio, defined as the segmented TLS area over the total tissue area, correlated with B lymphocyte levels and the expression of CXCL13, a chemokine associated with TLS formation, in 6140 patients spanning 16 tumor types from The Cancer Genome Atlas (TCGA). The prognostic models for overall survival indicated that the inclusion of the TLS ratio with TNM staging significantly enhanced the models' discriminative ability, outperforming the traditional models that solely incorporated TNM staging, in 10 out of 15 TCGA tumor types. Furthermore, when applied to biopsied treatment-naïve tumor samples, higher TLS ratios predicted a positive immunotherapy response across multiple cohorts, including specific therapies for esophageal squamous cell carcinoma, non-small cell lung cancer, and stomach adenocarcinoma. In conclusion, our deep learning-based approach offers an automated and reproducible method for TLS segmentation and quantification, highlighting its potential in predicting immunotherapy response and informing cancer prognosis.

The complete plastome genome sequence of Cynanchum otophyllum (Asclepiadaceae), a unique medicinal species in China.

Cynanchum otophyllum Schneid is an important medicinal plant in China. In this paper, the chloroplast genome of C. otophyllum was sequenced based on high-throughput technology, and the chloroplast genome structure characteristics and phylogenetic relationship of C. otophyllum were analyzed. The results showed the complete plastome genome size of C. otophyllumis 160,874bp, including one small single copy (SSC, 19,851bp) and one large single copy (LSC, 92,009bp) regions isolated by a pair of inverted repeat regions (IRs, 24,507bp). The whole plastome genome including 84 protein encoding genes, 8 rRNA and 37 tRNA. Based on the phylogenetic topologies, C. otophyllum shows close association with additional Gomphocarpus and Asclepias genus. This study contributes to an enhanced understanding of the genetic information of C. otophyllum and provides a theoretical basis for the development of molecular markers and phylogeographic of the species, as well as for constructing the phylogenetic tree of Asclepiadaceae.

Recombinant porcine Interferon-α and Interleukin-2 fusion protein (rPoIFNα+IL-2) shows potent anti-pseudorabies virus activity in vitro and in vivo.

Pseudorabies virus (PRV) variants have been widely prevalent since 2011, leading to substantial losses to the swine industry. Although PRV can cause cross-species transmission and induce human infection, no drugs can currently prevent PRV infection. Interferons (IFNs) and interleukin-2 (IL-2) are important cytokines that mediate several biological functions including antiviral activity and immune regulation. In this study, we expressed and purified a recombinant porcine IFN-α and IL-2 fusion protein (rPoIFNα+IL-2), which did not show a cytotoxic effect on PK-15 cells. The antiviral activity was evaluated in PK-15 cells using the cytopathic effect inhibition method, and the results indicated that rPoIFNα+IL-2 can inhibit the replication of PRV, with an antiviral activity of approximately 104 U/mL. Moreover, the proliferation of peripheral blood mononuclear cells was enhanced by rPoIFNα+IL-2. Additionally, rPoIFNα+IL-2 substantially increased the expression of IFN-stimulated genes, including IFIT1, ISG15, MX1, and OAS, which are critical for antiviral activity. Furthermore, rPoIFNα+IL-2 alleviated the clinical symptoms and reduced mortality in mice infected with PRV. Simultaneously, rPoIFNα+IL-2 increased the expression levels of IFN-γ and IL-10 and inhibited the expression of IL-1ß and IL-6. Additionally, the viral DNA copies in different tissues in the rPoIFNα+IL-2-treated group were lower than those in the untreated group. These findings indicate that rPoIFNα+IL-2 may serve as an antiviral agent for the prevention and treatment of PRV infection and may expand the potential function of IFN antiviral drugs in the future.

CRISPR-based targeted haplotype-resolved assembly of a megabase region.

Constructing high-quality haplotype-resolved genome assemblies has substantially improved the ability to detect and characterize genetic variants. A targeted approach providing readily access to the rich information from haplotype-resolved genome assemblies will be appealing to groups of basic researchers and medical scientists focused on specific genomic regions. Here, using the 4.5 megabase, notoriously difficult-to-assemble major histocompatibility complex (MHC) region as an example, we demonstrated an approach to construct haplotype-resolved assembly of the targeted genomic region with the CRISPR-based enrichment. Compared to the results from haplotype-resolved genome assembly, our targeted approach achieved comparable completeness and accuracy with reduced computing complexity, sequencing cost, as well as the amount of starting materials. Moreover, using the targeted assembled personal MHC haplotypes as the reference both improves the quantification accuracy for sequencing data and enables allele-specific functional genomics analyses of the MHC region. Given its highly efficient use of resources, our approach can greatly facilitate population genetic studies of targeted regions, and may pave a new way to elucidate the molecular mechanisms in disease etiology.

Recognition of Ionic Liquids as High-Voltage Electrolytes for Supercapacitors.

The electrochemical stability of electrolytes is essential to the working potential of supercapacitors. Ionic liquids (ILs) are being considered as safe alternatives to current organic electrolytes and attracting extensive interests owing to their inflammability, widened potential windows, and superior ionic conductivity. Novel supercapacitors with IL electrolytes exhibit attractive energy density and can be utilized in various energy storage systems. Most previous studies focused on electrochemical performances, while rare attentions were devoted to energy storage process details or mechanisms. This review comprehensively summarizes the latest progress on formulated IL electrolytes for different types of supercapacitors, with an emphasis on the intrinsic understanding of the related energy storage mechanisms. Subsequently, comparisons of various IL-based liquid-state electrolytes as well as the state-of-the-art advancements in optimizing ILs electrolytes are introduced. The authors attempt to reveal the inherent correlation between the usage of IL electrolytes and the properties of supercapacitors via referenced works. Some emerging applications of ionogel electrolytes based on conventional polymers and poly(IL)s for flexible supercapacitors are also presented, including the existing problems. In addition, challenges and future perspectives of research in this field are highlighted.

[L-arginine suppresses ischemia/reperfusion induced up-regulation of endothelin-1 production in a rat model of acute myocardial injury].

OBJECTIVE: To examine the level of endothelin-1 (ET-1) in serum and its expression in myocardium tissue during the development of acute myocardial ischemia/reperfusion (I/R) injury in rat and the effects of L-arginine (L-Arg) administration on these indexes. METHODS: One hundred and ten Wistar rats were randomly divided into nine groups to receive: (1) sham surgery, (2)ischemia (I, by ligation of anterior descending coronary artery for 30 minutes), (3) I+reperfusion (R, by the removal of the ligature) for 0.5 hour, (4) I+R for 1 hour, (5) I+R for 2 hours; group (6) ~ (9) also received I/R treatment as in group 2 ~ 5 respectively but with L-Arg pretreatment. Blood and myocardium tissue samples were collected by the end of the experiment for the analysis of: serum level of creatine kinase (CK), lactate dehydrogenase [LDH, by enzyme linked immunosorbent assay (ELISA)], ET-1 (radioimmunoassay), and the tissue content of ET-1 mRNA/peptide [by reverse-transcription polymerase chain-reaction (RT-PCR) and Western blotting]. RESULTS: In comparison with the sham treated control animals, the serum levels of CK, LDH, and ET-1 were all significantly higher in the groups treated with I/R (particularly those exposed to reperfusion). The myocardial tissue content of ET-1 mRNA/peptide were also significantly increased in I/R treated groups (particularly the I+R 2 hours group) as compared to control (ET-1 mRNA: 0.775 ± 0.029 vs. 0.310 ± 0.076; ET-1 peptide: 0.773 ± 0.055 vs. 0.340 ± 0.099, both P < 0.05). The i.v. administration of L-Arg significantly suppressed the up-regulation of tissue content of ET-1 mRNA /peptide in I/R treated animals (ET-1 mRNA: 0.340 ± 0.049 vs. 0.775 ± 0.029; ET-1 peptide: 0.390 ± 0.094 vs. 0.773 ± 0.055, both P < 0.05). CONCLUSION: L-Arg may be tested during certain stage of I/R injury as a therapeutic intervention for the suppression of ET-1 up-regulation.

[Application of nano-sized TiO2 photocatalysis to air purification and sterilization].

OBJECTIVE: To develop and evaluate the efficiency of air purification and sterilization instrument based on nano-sized TiO(2) photocatalytic technique. METHODS: The nano-sized TiO(2) photocatalytic air purification and sterilization instrument was designed and a sample had been prepared. The sterilization efficiencies for E.coli and Klebsiella by the nano-sized TiO(2) photocatalytic instrument and ultraviolet (UV) were measured in closed labs. The on-site efficiency of the instrument was evaluated, too. RESULTS: The nano-sized TiO(2) photocatalytic air purification and sterilization instrument was composed of five units: rough filter, nano-sized TiO(2) photocatalytic unit, activated carbon fiber filter, negative ion generator, and programmed control unit. The E.coli killing rates by the nano-sized TiO(2) photocatalytic instrument were 76.0%, 81.8%, 77.5%, and 80.7% at 30, 60, 90, and 120 minutes, respectively. There was no significant difference between the E.coli killing rates of the instrument and UV (P > 0.05), except the 120 minutes timepoint. The Klebsiella killing rates by the instrument were 78.4%, 79.5%, 67.3%, and 58.5% at 30, 60, 90, and 120 minutes, respectively. The Klebsiella killing efficiencies of the instrument at 30 and 60 minutes were better than that of UV (P < 0.01). There was no significant difference between the Klebsiella killing efficiencies of the instrument and UV (P > 0.05). CONCLUSION: The air sterilization efficiency of the nano-sized TiO(2) photocatalytic instrument should be equivalent or better as compared with the UV. This instrument might be used for the air purification and sterilization of the public locations.

[Multi-center phase II clinical trial of humanized anti-epidermal factor receptor monoclonal antibody h-R3 combined with radiotherapy for locoregionally advanced nasopharyngeal carcinoma].

OBJECTIVE: To evaluate the efficacy and safty of the humanized anti-epidermal factor receptor monoclonal antibody h-R3 in combination with radiotherapy for locoregionally advanced nasopharyngeal carcinoma. METHODS: Totally, 137 patients from 7 medical center around China were randomly divided into combined therapy group or control group. There was no difference in Karnofsky performance score between two groups. All patients in both groups received radical conventionally fractionated radiotherapy to the total dose of D(T) 70-76 Gy. For the combined therapy group, h-R3 was added at a dose of 100 mg i.v. weekly for 8 weeks started at the beginning of radiotherapy. RESULTS: Of the 137 eligilbe patients, 70 were in the combined therapy group treated by h-R3 plus radiotherapy and 67 in the control group by radiotherapy alone. The intent-to-treat (ITT) population consisted of 130 patients, while the per-protocol (PP) population was composed of 126 patients. The efficacy was assessed respectively at three point of time: the end of treatment, the 5th- and 17th-week after treatment. The complete response (CR) of the combined therapy group was significantly higher than that of the control group in both ITT and PP (ITT: 65.63%, 87.50%, 90.63% versus 27.27%, 42.42%, 51.52%; PP: 67.21%, 90.16%, 93.44% versus 27.69%, 43.08%, 52.31%; P < 0.05, respectively). The most common h-R3-related adverse reactions were fever (4.3%), hypotension (2.9%), nausea (1.4%), dizziness (2.9%) and rash (1.4%), which could be reversible if treated properly. Radiotherapy combined with 100 mg h-R3 i. v. weekly was tolerable and did not aggravate the side effects of radiation. The quality of life in the combined therapy group was comparable to that in the control group. CONCLUSION: This phase 1 multicenter clinical trial shows that h-R3 in combination with radiotherapy is effective and well-tolerated for the treatment of locoregionally advanced nasopharyngeal carcinoma.