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Abstract Background TCbHP (taxane + carboplatin + trastuzumab + pertuzumab) is the preferred neoadjuvant therapy regimen for human epidermal growth factor receptor 2 (HER2)-positive breast cancer. However, no consensus exists regarding whether specific populations may be exempt from carboplatin, allowing for de-escalation to the THP (taxane + trastuzumab + pertuzumab) regimen. Additionally, the optimal number of cycles for neoadjuvant THP remains unclear. We compared the efficacy and safety of neoadjuvant TCbHP and THP regimens, providing clinicians with a nuanced perspective to guide their treatment regimen selection. Methods This multicenter real-world study included patients with HER2-positive breast cancer undergoing neoadjuvant TCbHP or THP between March 2019 and February 2023. Efficacy was assessed through the pathological complete response (pCR) rate, while safety was evaluated through monitoring adverse events. Results Among 220 patients, 103 received 6 cycles of TCbHP (TCbHP×6), 83 received 6 cycles of THP (THP×6), and 34 received 4 cycles of THP (THP×4). The TCbHP×6 cohort exhibited a 66% pCR rate compared with 53% in the THP×6 cohort (P = 0.072). Subgroup analysis revealed that in patients aged ≤ 50 years, those with hormone receptor (HR)-negative status, and those with clinical stage T2, the pCR rate of the TCbHP×6 regimen was significantly higher than the THP×6 regimen (P 50, ≤60 years with HR-negative status or clinical stage T2-4. For patients in compromised general condition or lacking the specified indications, the THP×6 regimen emerges as a lower-toxicity alternative with satisfactory efficacy. To ensure treatment efficacy, a minimum of 6 cycles of neoadjuvant THP is required.

Thamnaconus septentrionali has excellent breeding characteristics, is omnivorous, easy to domesticate, is especially suitable for netting, cleans the netting, and effectively reduces labor-costs. T. septentrionali is available in various sizes and is raised in the market at 100 g. Under water temperature of 18–25 ℃, after 5–6 months of breeding, it reaches commercial size. As the Yellow Sea and Bohai Sea are cold in winter, a suitable breeding cycle for green fins is from May to November annually. Under natural conditions, the spawning period of T. septentrionali in the Yellow Sea and Bohai Sea is from early May to early June, making good use of the suitable breeding cycle of net box culture. Therefore, reproductive regulation is required to advance the reproductive period of T. septentrionali. Previous studies have primarily focused on the expression and functional analysis of individual genes. In contrast, no studies exist on the overall expression analysis of gonad-related genes based on histology. Herein, we conducted the first transcriptome sequencing analysis of the spermatophores and ovaries of T. septentrionali using an Illumina high-throughput sequencing platform. Overall, 165, 981, 523 raw reads were sequenced from the cDNA, and 161, 234, 846 clean reads were obtained after quality control. The Q20 of each sample was > 98.43%, Q30 was > 95.25%, and GC content of the sample bases was greater than 52.31%. Our findings indicate that the sequencing data are accurate, of good quality, and were used for subsequent analyses. The obtained unigenes were annotated in the NR (NCBI non-redundant protein sequences), NT (NCBI nucleotide sequences), PFAM (protein family), KOG (euKaryotic Ortholog Groups), SwissProt (a manually annotated and reviewed protein sequence database), GO (Gene Ontology), and KO (KEGG Orthology) databases, and 24, 009, 35, 057, 18, 453, 26, 971, 30, 294, 11, 420, and 21, 613 unigenes were annotated, respectively. The KEGG annotation results were divided into five major categories: organic systems, with 2, 115 genes in nine pathways; metabolism, with 1, 444 genes in 12 pathways; environmental information processing, with 1, 200 genes in three pathways; and genetic information processing, with 1, 128 genes in four pathways. Environmental information processing (1, 200 genes in three pathways), genetic information processing (1, 128 genes in four pathways), and cellular processes (1, 128 genes in four pathways). The number of genes enriched in cellular process-related pathways was 927 distributed in four related pathways. The signal transduction pathway was the most annotated KEGG pathway with 804 genes. The experiment used 24, 546 unigenes annotated in the KOG database, which were then classified into 26 categories based on their function. The largest number of unigenes were annotated in the signal transduction mechanism category (5, 411), followed by the general function prediction-only category (4, 252 unigenes), and posttranslational modification, protein folding and chaperone category (1, 713 unigenes). Unigene posttranslational modification, protein turnover, chaperones; transcription, 1, 613; and other related functions. There were 18, 954 differentially expressed genes (DEGs) in the spermatophore and ovary transcriptomes of the T. septentrionali, with 11, 265 genes up-regulated and 7, 689 down-regulated in the ovary relative to the spermatophore. The GO functional enrichment analysis revealed that DEGs were most enriched in the cellular component of biological processes, intracellular component of the cellular component subclass, and nucleic acid binding of the molecular functional subclass. GO analysis of the DEGs in males and females provided a partial list of genes associated with the reproductive process (GO: 0022414), sexual reproduction (GO: 0019953), gamete formation (GO: 0007276,), sex differentiation (GO: 0007548), and gonad development (GO: 00084060). To further characterize the specific functions of the DEGs in the spermatophores and ovaries of T. septentrionali, the enriched signaling pathways were further analyzed using the KEGG database. In total, 154 KEGG pathways were included, of which the top 30 were selected. The functional pathways with the highest number of expressed genes were closely associated with gonadal sex differentiation and development, including the insulin signaling pathway, steroid hormone biosynthesis, FoxO signaling pathway, M-TOR signaling pathway, and progesterone-mediated oocyte maturation. Among these, the FoxO signaling pathway regulatory genes are expressed in multiple processes, such as cell cycle control, apoptosis, and gluconeogenesis. The insulin signaling pathway plays an important role in regulating developmental, metabolic, and lifespan physiological processes, and the insulin signaling pathway is involved in gonadal development and maturation. Nine DEGs, bmp2, sox3, figla, hsd17b1, cyp19a, cyp17, foxl2, star, and amh, were selected for real-time fluorescence quantitative PCR (qRT-PCR) validation. The qPCR results were consistent with those of RNA-seq analysis. GO and KEGG enrichment results were analyzed and revealed that amh, cyp17, and star are imperative in male spermatogenesis of T. septentrionali; bmp2, foxl2, cyp19a, figla, and hsd17b1 are essential in female oogenesis and ovarian steroidogenesis. By comparing the transcriptome expression differences between the spermatophore and ovary of T. septentrionali, we elucidated the gene expression characteristics of the spermatophore and ovary, laying the foundation for future research on the mechanism of reproductive development of T. septentrionali and providing theoretical support for optimizing reproductive regulation techniques.

Fish skin is a multipurpose tissue with important functions—protection, perception, and hormone metabolism—in the fish body. As a byproduct of fish processing, it can be used as a material for extracting collagen, making leather, and related pathological research. However, there have been few reports on the tissue structure of fish skin. This study aimed to analyze the structural characteristics of Thamnaconus septentrionalis fish skin in order to provide a reference for the utilization of its resources. Six T. septentrionalis samples were selected and their skins were observed using frozen sectioning, skeletal, hematoxylin & eosin (H & E), Periodic acid-Schiff (PAS), Masson, Van Gieson (VG) staining, and scanning electron microscopy. The images obtained by slice scanning were analyzed using SlideViewer (3DHISTECH), ImageJ, and Photoshop software. The SEM images were analyzed using ImageJ and Photoshop software. SPSS25.0 software was used to analyze the data of epidermal and squamous layers, conical bone bulge, basal layer, and dermis thicknesses. The results showed that the histological structure of the skin comprised four parts: epidermis, scale layer, dermis, and subcutaneous tissue. The thickness of the epidermal layer was (26.81±7.48) μm. This layer was mainly composed of epithelial and basal cells but also contained a large number of mucus cells. The scale layer consisted of the spine of conical bone and a substrate. The thickness of this layer was (22.49±5.19) μm, and there were two to four rows of the spine of conical bone with different diameters, different bending degrees at the top, and a height of (257.13±10.41) μm unevenly distributed on the base plate. The average thickness of the dermis was (176.97±21.11) μm, and the dermis thickness of the head was lower than that of the middle part of the body, but higher than that of the tail, mainly composed of collagen fiber. The subcutaneous tissue layer was mainly comprised of collagen fibers and non-fibrous stroma. In this study, the histological pattern of the skin of T. septentrionalis was mapped according to its characteristics, providing valuable information for fish skin resource usage.

BackgroundPrimary hepatic neuroendocrine tumors (PHNETs) are an utterly rare entity. The diagnosis of PHNETs could legitimize when an extrahepatic primary NET must always be excluded. PHNETs can achieve a high survival rate after complete surgical resection, however, most patients still have an 18% risk of recurrence within 5 years after surgery. In our case, the recurrence occurred 8 years after the first hepatectomy, which is relatively rare in the current literature. Therefore, rigorous postoperative follow-up is necessary for early detection and timely treatment of recurrent PHNETs.Case informationWe report a case of PHNET in a 24-year-old previously healthy female patient who relapsed 8 years after hepatectomy. This case focuses on the importance of diagnosis of primary and recurrent PHNETS in young patients, rare pathological types, and post-operative follow-up.ConclusionThis case report detailed the rare pathological morphology and characteristic immunohistochemical markers in our case for PHNETS, which enhanced the new understanding of the diagnosis of this entity. In addition, we also highlighted the variable duration of recurrence after treatment of PHNETs. The 8-year recurrent period in our case suggests the importance of regular examination in patients with PHNETs by following the doctor’s instructions.

Background and objective Tumor microenvironment (TME) is one of the important factors in tumorigenesis and progression, in which tumor-associated macrophages (TAMs) play an important role in non-small cell lung cancer (NSCLC) progression. However, the mechanism of TAMs in NSCLC progression remains unclear, so this study aimed to investigate the role of TAMs in NSCLC progression and to find potential therapeutic targets. Methods Gene Expression Profiling Interactive Analysis (GEPIA) database was used to analyze the expression of prostaglandin E2 receptor 4 (EP4) mRNA in NSCLC and normal lung tissues; the protein expression levels of cyclooxygenase-2 (COX-2), EP4, cluster of differentiation 86 (CD86), CD163 and CD31 were detected by immunohistochemistry (IHC) in 120 NSCLC tissues and 24 paracancerous tissues specimens. The nude mouse lung adenocarcinoma cell A549 and macrophage RAW264.7 co-transplanted tumor model was established. And the samples were collected by gavage with EP4 inhibitor E7046, and then stained with hematoxylin-eosin (HE), IHC, and immunofluorescence (IF), and then detected by Western blot for the epithelial mesenchymal transformation (EMT) of the tumor tissues of the nude mice in each group. Western blot was used to detect the expressions of EMT related protiens in each group of nude mice; full-length transcriptome sequencing was used to screen the key genes causing liver metastasis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed. Results EP4 mRNA expression level in NSCLC tissues was generally lower than that in normal lung tissues (P0.05); the main pathways enriched in the differential genes of the full-length transcriptome were the PI3K-AKT and MAPK signaling pathways. Conclusion During NSCLC development, the COX-2/PGE2/EP4 axis may promote tumor progression by inducing macrophage functional activation, and EP4 may be a potential new target for tumor immunotherapy. This study provides new perspectives and ideas for in-depth exploration of the mechanisms of NSCLC development, as well as a theoretical basis for the development of new therapeutic strategies for NSCLC.

The morphological characteristics, ecological habits, and physiological structures of fish vary rapidly in the early growth and development stage. Understanding the changes in the postembryonic early development morphology of organs or tissues could provide basic information for the classification or identification of fish and the investigation or utilization of early resources. Morphological characteristic observation is one of the most commonly used methods. The existing studies on the early development of a certain trait in the late larval and juvenile stage are unclear. Most of the digital images are side views, resulting in a lack of dorsal views, ventral views, or local images, which is not conducive to the understanding of larval and juvenile. In this study, the variations in observable, measurable, and countable traits in postembryonic early development of Thamnaconus septentrionalis were explored using optical microscopy and stereomicroscopy. The observable traits included the developmental changes of yolk sac, scales, and notochord curvature. The measurable traits included the growth changes of total length, body length, body height, and oral fissure. The countable characters included the number of fins in the second dorsal fin, anal fin, caudal fin, and the number of pterygiophore. The results showed that at (23.0±1.0) ℃, the newly hatched larvae moved up and down in the water, with a phenomenon of convergent clustering under light irradiation; the pectoral fin appeared at 1 d, and the heart rate was approximately 50 beats/min; the optic capsule was completely black, the maxillar appeared, the oral fissure extended below the eyes, the yolk sac disappeared, and the first dorsal fin base appeared at 4 d; the girdle was single and colorless and began from the front edge of the belly, the oil globule disappeared, and the pelvic fin base appeared at 7 d; the first dorsal fin base and pelvic fin base broke membrane at 8 d; the first dorsal fin and pelvic fin were significantly longer than before, transparent rounded scales were observed at 9 d; swimming ability was improved, indicated by hovering back and forth gnawing pool wall, transversally conical barbs appeared at the base of the first dorsal fin at 11 d; the swim bladder and postcleithrum on both sides of the belly were clearly visible at 12 d; the second dorsal fin and anal fin pterygiophore appeared at 14 d, the second dorsal fin pterygiophore spans seven sarcomeres and the anal fin pterygiophore spans six sarcomeres. There are 10~12 melanin masses in the second dorsal fin and 5~6 in the anal fin, the conical teeth appeared on the upper and lower jaws at 16 d; the second dorsal fin and anal fin were differentiated at 19 d; the swim bladder fills about 25% of the abdominal cavity; notochord curves upward clearly and the stalk of caudal fin came out at 20 d; the fin strips of the second dorsal fin, anal fin, and caudal fin were well-developed at 24 d; the open mouth was round and changed from swallowing to sucking at 30 d, and the rainbow cells were colorful and the body surface near the gill was the most dense; the abdominal cavity has a strong metallic texture, the hard spine of the pelvic fin was degraded, close to the girdle at 39 d; 4~5 black bands were arranged regularly on the body, and the lateral line was consistent with the vertebral column at 50 d. The R2 values of the curve equations of measurable traits were all greater than 0.90. The height of the first dorsal fin increased exponentially 4~30 d. The visual sac increased linearly from 0~11 d to 11~20 d, and the increment of 0~11 d was less than that of 11~20 d, while the increment of 21~30 d was exponential. The oral fissure in 4~13 d and 22~30 d increased linearly, while that in 13~22 d decreased linearly. The total length, body length, and body height increased linearly before the inflection point and exponentially after the inflection point, which was concentrated in 19~23 d. The number of fin pterygiophore of the second dorsal fin was 31~36, while that in the anal fin was 33~35. The number of pectoral fins and caudal fins were 15~18 and 12, respectively. In conclusion, metamorphosis occurs in the postembryonic early development of T. septentrionalis, showing an allometric growth pattern, with a slow linear growth in the rotifer-feeding stage and a fast exponential growth in the artemia-feeding stage. The changes in the first dorsal fin and pelvic fin were correlated with age in days. The changes in the second dorsal and anal fins were correlated with the total length. This study enriches the early biological theory of T. septentrionalis and provides basic data for the investigation or utilization of its seedling breeding and resources. The partial enlarged detail is helpful for scholars to further understand the larval or juvenile fish and perform deep research.

Abstract Background and Objective Tebipenem pivoxil (TP) is a carbapenem and is applied against pneumonia, otitis media, and sinusitis. This study compared the pharmacokinetics (PK) and safety of a test (T) preparation and reference (R) preparation of TP in healthy Chinese adults. Methods This study was a single-center, randomized, open, single-dose (fasting/postprandial) oral administration, two-agent, two-sequence, two-cycle, crossover bioequivalence trial. A total of 60 participants were enrolled (24 fasting and 36 postprandial). All participants were randomly assigned to the TR sequence and RT sequence. Subsequently, they switched T sequences or R sequences 7 days later. PK blood samples were collected according to the protocol, plasma TP concentration was determined by liquid chromatography-mass spectrometry, main PK parameters were calculated based on a non-compartment model, and adverse events were recorded during the test. Results In the feeding arm, the geometric mean ratio of maximum concentration (C max) was 89.84% (90% confidence interval 84.33–95.70), the geometric mean ratio of area under the plasma concentration–time curve from time 0 to last time of quantifiable concentration (AUC0–t ) was 86.80% (83.62–90.10), and the geometric mean ratio of area under the plasma concentration–time curve from time 0 to infinity time of quantifiable concentration (AUC0–∞) was 86.90% (83.73–90.20), which were within the acceptable range of bioequivalence (80–125%). In the fasting arm, the geometric mean ratio of C max was 96.07% (89.62–102.99), the geometric mean ratio of AUC0–t was 93.09% (90.47–95.78), and the geometric mean ratio of AUC0–∞ was 93.09% (90.48–95.77), which was within the acceptable range of bioequivalence (80–125%). Hence, the T preparation and R preparation of TP had bioequivalence in the fasting arm and feeding arm of the clinical trial. In addition, all adverse events were mild, and no severe adverse events were noted. Conclusion Preparations T and R of TP were bioequivalent in the fasting and postprandial groups in clinical trials, and TP was safe.

Abstract Lung cancer, specifically the histological subtype lung adenocarcinoma (LUAD), has the highest global occurrence and fatality rate. Extensive research has indicated that RNA alterations encompassing m6A, m5C, and m1A contribute actively to tumorigenesis, drug resistance, and immunotherapy responses in LUAD. Nevertheless, the absence of a dependable predictive model based on m6A/m5C/m1A-associated genes hinders accurately predicting the prognosis of patients diagnosed with LUAD. In this study, we collected patient data from The Cancer Genome Atlas (TCGA) and identified genes related to m6A/m5C/m1A modifications using the GeneCards database. The “ConsensusClusterPlus” R package was used to produce molecular subtypes by utilizing genes relevant to m6A/m5C/m1A identified through differential expression and univariate Cox analyses. An independent prognostic factor was identified by constructing a prognostic signature comprising six genes (SNHG12, PABPC1, IGF2BP1, FOXM1, CBFA2T3, and CASC8). Poor overall survival and elevated expression of human leukocyte antigens and immune checkpoints were correlated with higher risk scores. We examined the associations between the sets of genes regulated by m6A/m5C/m1A and the risk model, as well as the immune cell infiltration, using algorithms such as ESTIMATE, CIBERSORT, TIMER, ssGSEA, and exclusion (TIDE). Moreover, we compared tumor stemness indices (TSIs) by considering the molecular subtypes related to m6A/m5C/m1A and risk signatures. Analyses were performed based on the risk signature, including stratification, somatic mutation analysis, nomogram construction, chemotherapeutic response prediction, and small-molecule drug prediction. In summary, we developed a prognostic signature consisting of six genes that have the potential for prognostication in patients with LUAD and the design of personalized treatments that could provide new versions of personalized management for these patients.

Abstract The tumor microenvironment and cancer-associated fibroblasts (CAFs) play crucial roles in tumor development, and their metabolic coupling remains unclear. Clinical data showed a positive correlation between PDGF-BB, CAFs, and glycolysis in the tumor microenvironment of oral tongue squamous cell carcinoma patients. In vitro, CAFs are derived from hOMF cells treated with PDGF-BB, which induces their formation and promotes aerobic glycolysis. Mitophagy increased the PDGF-BB-induced formation of CAF phenotypes and aerobic glycolysis, while autophagy inhibition blocked PDGF-BB-induced effects. Downregulation of miR-26a-5p was observed in CAFs; upregulation of miR-26a-5p inhibited the expression of mitophagy-related proteins ULKI, Parkin, PINK1, and LC3 and aerobic glycolysis in PDGF-BB-induced CAFs. PDGF-BB-induced CAFs promoted tumor cell proliferation, invasion, metastasis, NF-κB signaling pathway activation, and PDGF-BB secretion. Thus, PDGF-BB is associated with lactate-induced CAF formation and glucose metabolism reprogramming. These findings indicate potential therapeutic targets in oral tongue squamous cell carcinoma.

Background and objective Currently, a significant number of miners are involved in mining operations at the Gejiu tin mine in Yunnan. This occupational setting is associated with exposure to dust particles, heavy metals, polycyclic aromatic hydrocarbons, and radioactive radon, thereby significantly elevating the risk of lung cancer. This study aims to investigate the involvement of leptin-mediated extracellular regulated protein kinase (ERK) signaling pathway in the malignant transformation of rat alveolar type II epithelial cells induced by Yunnan tin mine dust. Methods Immortalized rat alveolar cells type II (RLE-6TN) cells were infected with Yunnan tin mine dust at a concentration of 200 μg/mL for nine consecutive generations to establish the infected cell model, which was named R200 cells. The cells were cultured normally, named as R cells. The expression of leptin receptor in both cell groups was detected using the Western blot method. The optimal concentration of leptin and mitogen-activated protein kinase kinase (MEK) inhibitor (U0126) on R200 cells was determined using the MTT method. Starting from the 20th generation, the cells in the R group were co-cultured with leptin, while the cells in the R200 group were co-cultured with the MEK inhibitor U0126. The morphological alterations of the cells in each group were visualized utilizing hematoxylin-eosin staining. Additionally, concanavalin A (ConA) was utilized to detect any morphological differences, and an anchorage-independent growth assay was conducted to assess the malignant transformation of the cells. The changes in the ERK signaling pathway in epithelial cells after the action of leptin were detected using the Western blot method. Results Both the cells in the R group and R200 group express leptin receptor OB-R. Compared to the R200 group, the concentration of leptin at 100 ng/mL shows the most significant pro-proliferation effect. The proliferation of R200 cells infected with the virus is inhibited by 30 μmol/L U0126, and a statistically significant divergence was seen when compared to the control group (P

The East Asian common octopus (Octopus sinensis), is mainly distributed along the southeast coast of China. It has become an important aquaculture species in the coastal areas of the Fujian and Guangdong province due to its excellent culture features such as high conversion rate, fast growth rate, large size, and high market price. The main culturing methods of O. sinensis are industrialized, indoor, and cage culturing. The density is high during cage culturing. If the feed is insufficient or the feeding interval too long, there are attacks and cannibalism among individuals, which adversely impacts the culturing income. Therefore we urgently need to undertake digestive physiology research to explore the digestive functions of O. sinensis. This research will provide valuable information to identify an appropriate feeding frequency and reduce the economic loss caused by cannibalism. At present, there is no relevant research on the digestive physiology of O. sinensis. In this study, the histological structure of each digestive organ of O. sinensis was observed to explore the relationship between the tissue structure and digestive function. We investigated the trypsin, amylase, and lipase activities in the anterior salivary gland, posterior salivary gland, crop, digestive gland, stomach, cecum, and intestine before and after feeding. The changes in plasma glucose, plasma triglyceride, plasma total cholesterol, plasma total protein and muscle glycogen before and after feeding were analyzed to determine the digestion and absorption duration in O. sinensis.O. sinensis is an aggressive carnivore with a highly developed digestive system. The histological results showed that the crop, stomach, cecum, and intestine of O. sinensis is composed of three layers, the mucosal, submucosal, and muscular, and the inner wall contained many folds. Both the anterior and posterior salivary glands were compound tubular glands, which were composed of oval and circular gland tubes with many secretory ducts. The crop widens and expands from the esophagus into a large lumen, and the cuticle of the mucosal layer was thin (with a thickness of approximately 2.57–5.44 μm). The stomach was spherical, the gastric cavity was biased to one side, and the inner wall contained many folds. The muscular layer of the gastric wall was the most developed layer with the cuticle approximately 99.97–383.82 μm. The cecum was double helix and the inner wall contained rich long folds and secondary folds. The submucosa at the free end of some of the long lateral folds contained some mucous glands, and the mucosal layer without any cuticle. The digestive gland was the largest proportion of the digestive system and was composed of hepatic lobules. The boundary of the hepatic lobules was difficult to distinguish. The diameter of the intestine was approximately 2.50 mm, the tube wall was thin and the mucosal layer had no cuticle. The intestinal mucosa forms wavy longitudinal folds in the mucosa, including a pair of longer folds, which occupied almost the entire intestine. The analysis of the digestive enzyme activities and nutrient metabolism showed that trypsin activity was significantly higher than that of lipase and amylase. The trypsin activity in the digestive gland reached (148.74±21.25) U/mg, and the amylase activity in the digestive gland and stomach reached (3.68±0.59) U/mg and (2.48±0.64) U/mg, respectively. The lipase activities did not vary between the different tissues. After feeding, the plasma total protein concentration reached the lowest levels of (161.50±67.51) mg/mL at 120 min, and then increased. The plasma glucose and muscle glycogen concentrations of O. sinensis reached maximums of (1.54±0.44) mmol/L and (4.75± 0.13) mg/g at 200 min, respectively. Feeding had little effect on the plasma lipid concentration in O. sinensis, the plasma cholesterol concentration did not vary significantly while feeding (P < 0.05), and there was limited variation in the plasma triglyceride concentration. The digestion process was divided into two stages. The first stage occurs in 0~60 min as food entered the crop and stomach, and the digestive gland secreted large doses of trypsin. Initially, the trypsin in the stomach mixed with the food, resulting in decreased trypsin activity. At this stage, only small amounts of nutrients are absorbed and the body needs to consume nutrients to supply energy for the digestion process. Therefore, a decrease in plasma glucose, plasma protein, and muscle glycogen content occurred. The second stage occurs in 120~300 min and involves high trypsin activity. In the progress of extracellular digestion and intracellular digestion, the trypsin activity of various digestive organs decreased gradually, and nutrients increased gradually. At 400 min, the trypsin activity, plasma glucose, plasma protein, and muscle glycogen returned to the pre-feeding levels, indicating the end of the digestion and absorption process. The whole digestion and absorption process of O. sinensis lasted approximately 400 min. Therefore, the appropriate feeding interval for O. sinensis culture is 6~7 h.

Abstract Aims This study aims to explore the role of exosomes from cancer-associated fibroblasts (CAFs) induced by PDGF-BB in promoting the malignancy of oral squamous cell carcinoma (OSCC) and provide new insight into the mechanism of OSCC progression and its treatment. Main methods Exosomes were extracted from human oral mucosa fibroblasts (hOMFs) and CAFs. Differentially expressed miRNAs of exosomes between hOMFs and CAFs were analysed using high-throughput sequencing and self-programmed R software. Cal-27, a human tongue squamous carcinoma cell line, was treated with exosomes. Differentially expressed miRNAs between clinical cancer tissues and adjacent tissues and between hOMF and CAF exosomes were verified by qRT‒PCR. The effect of miR-3529-3p on Cal-27 cells was clarified by overexpressing or knocking down miR-3529-3p in Cal-27 cells. Sample expression and differentially expressed miRNA expression were compared between cancer and paracarcinoma tissues. Key findings We found that exosomes from CAFs (CAF-Exos) were internalized by tongue squamous carcinoma cells and promoted their proliferation, migration, invasion, and antiapoptotic effects. MiR-3529-3p was a significant differentially expressed miRNA between CAF-Exos and exosomes from hOMFs (hOMF-Exos). The overexpression of miR-3529-3p promoted proliferation, migration, and invasion and inhibited apoptosis of Cal-27 cells. Significance This study explores the role of PDGF-BB-induced CAFs in promoting malignancy in OSCC. This study will provide new insight into the mechanism of OSCC progression and its treatment.

Background: Both CellSearch and CellCollector have been accepted as the proper devices to capture CTC by domestic approval department. However, there is little article about the comparison between these two devices around the world. Herein, we conducted the real-world study to compare with these two devices and to re-verify the efficacy of CTC counts. Methods: Patients who meet the following points should be included in the analysis. 1. Female, aged 18 years or older; 2. Eastern Cooperative Oncology Group (ECOG) score 0–2; 3. With at least one measurable tumor lesion; 4. Clear immunohistochemistry result; 5. Accept at least one CTC test. Patients were excluded in the analysis if they had a history of malignant tumors, incomplete follow-up information. Results: 536 metastatic breast cancer patients who had been detected for CTC at least once by CellSearch or CellCollector were included in the analysis. CellCollector in vivo CTC detection technology has a higher detection rate than the CellSearch system (69.2% vs 57.4%, P = 0.009). However, the proportion of CTC≥5 detected by CellSearch was higher than CellCollector (37.4% vs 16.3%, P

The acute toxicity of cadmium (Cd2+) to juvenile Thamnaconus septentrionalis was determined using a hydrostatic biological assay. According to the pre-experiment results, four Cd2+ concentration gradient test groups (8.19, 9.18, 10.30, and 11.56 mg/L) and one blank control group were selected for the acute toxicity test using the equilogarithmic spacing method. The mortality, LC50, and safe concentration of Cd2+ in juvenile fish at 24 h, 48 h, 72 h, and 96 h were calculated. Based on the experimental results of acute toxicity of Cd2+ to juvenile Thamnaconus septentrionalis, the Cd2+ concentration gradients were set as 1.84 mg/L, 2.76 mg/L, 3.68 mg/L and 4.60 mg/L at the safe concentration multiple. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione antioxidant enzyme (GSH-PX), and the content of malondialdehyde (MDA) in liver and gill tissues were observed at 6 h, 12 h, 24 h, 48 h, 72 h and 96 h. The results showed that as Cd2+ concentration increased, the acute toxicity gradually increased, the LC50 at 24 h, 48 h, 72 h and 96 h was 11.47 mg/L, 10.82 mg/L, 9.84 mg/L, and 9.19 mg/L, respectively, and the safe concentration of Cd2+ on juvenile filefish was 0.92 mg/L at 96 h. Within 6 h, SOD activity in each concentration group was significantly higher than that in the control group (P < 0.01); SOD activity first decreased and then increased within 6–24 h; however, from 24 h to 96 h, SOD activity of all concentration groups showed a downward trend. SOD activity in the 1.84 mg/L and 2.76 mg/L groups was always higher than that in the control group within 96 h. The enzyme activities of the 3.68 mg/L and 4.60 mg/L groups decreased to below that of the control group from 6 h to 12 h, and increased at 24 h, after which it showed a constant downward trend and remained lower than that of the control group. From 6–12 h, MDA content first decreased and then increased. At 24 h, the enzyme content in each treatment group decreased, but was still higher than that in the control group, and the difference was significant (P < 0.05). From 12 h to 48 h, the 1.84 mg/L and 2.76 mg/L groups showed a trend of first decreasing and then increasing, whereas within the 24 h to 96 h period, the MDA content in the 3.68 mg/L and 4.60 mg/L groups kept increasing, and was significantly higher than in the control group (P < 0.01). At 6 h, CAT activity in all treatment groups was significantly increased when compared to that of the control group; however, CAT activity in the 3.68 mg/L and 4.60 mg/L groups was significantly inhibited at 48–96 h. Overall, the activity of GSH-PX in each treatment group showed a decreasing, increasing, and then decreasing trend within 6–96 h. The activity of GSH-PX in the 1.84 mg/L group was higher than that in the control group within 96 h. When compared with the control group, the activity of the 2.76 mg/L and 3.68 mg/L groups decreased from 24 h to 96 h, and the longer the stress time, the lower the activity. In the highest concentration group (4.60 mg/L), the enzyme activity first increased and then decreased, and the GSH-PX activity decreased to 300.12 U/mg prot at 96 h, which was significantly different from that in the control group (P < 0.01). In the 1.84 mg/L group, there was no significant change in liver tissue within 72 h, but in the 1.84 mg/L group at 96 h and the 2.76 mg/L group at 24 h, the cell volume was slightly increased, cells had an irregular shape, and some cell membrane boundaries were blurred. At 48–96 h in the 2.76 mg/L group, and 24–48 h in the 3.68 mg/L and 4.60 mg/L groups, the liver cells were disordered and scattered, and the structure of liver cells was mostly incomplete. After 72–96 h in the 3.68 mg/L group and 72 h in the 4.60 mg/L group, the abnormality of liver cells was significantly aggravated, including cell hypertrophy, large area disintegration, cytoplasm overflow, and cell cavitation. At 96 h in the 4.60 mg/L group, the nucleus shrank, intracellular material gathered, and a large irregular blank appeared. For 72 h in the 1.84 mg/L group and 24 h in the 2.76 mg/L group, there was no significant change in gill tissue structure, whereas after 96 h in the 1.84 mg/L group, 48 h in the 2.76 mg/L group, and 24 h in the 4.60 mg/L group, the gill microplates were curved and club-like with mucus secreted around them, and the epithelial cells were enlarged, showed edema and partial hyperplasia, and the distance between the adjacent lamella became smaller. At 72 h in the 2.76 mg/L group, and at 24–48 h in the 3.68 mg/L and 4.60 mg/L groups, a small number of adjacent gill lamellae would fuse and form a small epithelial cell plate, with a large amount of mucus around them, and the length of gill lamellae was significantly shortened. In the 2.76 mg/L group at 96 h and the 3.68 mg/L group at 72 h, most of the basal lamellae had fused together, the epithelial cells of the gill lamellae were swollen and showed hyperplasia, and exfoliated epithelial cells were scattered around. At 96 h in the 3.68 mg/L group and 72–96 h in the 4.60 mg/L group, adjacent gill lamellae had adhered and fused with each other without a free end, and a large number of cells were necrotic and exfoliated.

Abstract Background Cancer-associated fibroblasts (CAFs) have significant tumor regulatory functions, and CAFs-derived exosomes (CAFs-Exo) released from CAFs play an important role in the progression of oral squamous cell carcinoma (OSCC). However, a lack of comprehensive molecular biological analysis leaves the regulatory mechanisms of CAFs-Exo in OSCC unclear. Methods We used platelet derived growth factor-BB (PDGF-BB) to induce the transformation of human oral mucosa fibroblast (hOMF) into CAFs, and extracted exosomes from the supernatant of CAFs and hOMF. We validated the effect of CAFs-Exo on tumor progression by exosomes co-culture with Cal-27 and tumor-forming in nude mice. The cellular and exosomal transcriptomes were sequenced, and immune regulatory genes were screened and validated using mRNA-miRNA interaction network analysis in combination with publicly available databases. Results The results showed that CAFs-Exo had a stronger ability to promote OSCC proliferation and was associated with immunosuppression. We discovered that the presence of immune-related genes in CAFs-Exo may regulate the expression of PIGR, CD81, UACA, and PTTG1IP in Cal-27 by analyzing CAFs-Exo sequencing data and publicly available TCGA data. This may account for the ability of CAFs-Exo to exert immunomodulation and promote OSCC proliferation. Conclusions CAFs-Exo was found to be involved in tumor immune regulation through hsa-miR-139-5p, ACTR2 and EIF6, while PIGR, CD81, UACA and PTTG1IP may be potentially effective targets for the treatment of OSCC in the future.

Abstract. Background:. Endocrine therapy (ET) and ET-based regimens are the preferred first-line treatment options for hormone receptor (HR)-positive and human epidermal growth factor receptor 2 (HER2)-negative metastatic breast cancer (HR+/HER2– MBC), while chemotherapy (CT) is commonly used in clinical practice. The aim of this study was to investigate the efficacy and clinical outcome of ET and CT as first-line treatment in Chinese patients with HR+/HER2– MBC. Methods:. Patients diagnosed with HR+/HER2–MBC between January 1st, 1996 and September 30th, 2018 were screened from the Chinese Society of Clinical Oncology Breast Cancer database. The initial and maintenance first-line treatment, progression-free survival (PFS), and overall survival (OS) were analyzed. Results:. Among the 1877 included patients, 1215 (64.7%) received CT and 662 (35.3%) received ET as initial first-line treatment. There were no statistically significant differences in PFS and OS between patients receiving ET and CT as initial first-line treatment in the total population (PFS: 12.0 vs. 11.0 months, P = 0.22; OS: 54.0 vs. 49.0 months, P =0.09) and propensity score matched population. For patients without disease progression after at least 3 months of initial therapy, maintenance ET following initial CT (CT-ET cohort, n = 449) and continuous schedule of ET (ET cohort, n = 527) had longer PFS than continuous schedule of CT (CT cohort, n = 406) in the total population (CT-ET cohort vs. CT cohort: 17.0 vs. 8.5 months; P

The greenfin horse-faced filefish (Thamnaconus septentrionalis) and rockfish (Sebastes schlegeli) occupy important positions in the offshore net fishery in Shandong Province. Interest in their mariculture has been developing rapidly in recent years as candidates for submerged cage open-sea aquaculture. With the development of breeding techniques and the expansion of large- scale farming, fish disease may become a serious constraint that limits sustainable aquaculture and leads to great economic losses. Epidemiological investigation is the basis of disease control and should be carried out throughout the culture process. In this study, we describe the diseases of T. septentrionalis and S. schlegeli caused by Aeromonas salmonicida subsp. masoucida. In November 2018, an outbreak of T. septentrionalis disease was observed in a farm located in Penglai, Shandong Province, and an outbreak of S. schlegeli disease occurred in the same farm in April 2019, with daily mortalities of 0.4%~1% and about 1%, respectively. The main symptoms in the diseased fish were ulcers, redness, swelling, and bleeding in the mouth. Most diseased fish in the ponds showed "red mouth". No parasites were observed by the naked eye or light microscope. From the liver, spleen, and kidney of all the diseased fish, many homogeneous colonies were observed after three days incubation on TSA and 2216E agar plates. All strains had the same shape, color, and size, and the 16S rRNA genes of all strains were the same, with high identity with A. salmonicida. The virulence of the isolates was tested experimentally via injection with T. septentrionalis (infected by 2018TS-1) and S. schlegeli (infected by 2019SS-1) in the laboratory to calculate the median lethal dose (LD50). The results showed that the LD50 of 2018TS-1 to T. septentrionalis was 1.78×105 CFU/fish, and that of 2019SS-1 to S. schlegeli was 0.89×105 CFU/fish. The dead fish of the experimentally infected group showed ulcers and red mouth, the same symptoms as in naturally infected fish. Dominant colonies isolated from experimentally infected fish were all identified as A. salmonicida by 16S rRNA gene sequencing, which indicated that 2018TS-1 and 2019SS-1 were the pathogens of T. septentrionalis and S. schlegeli, respectively. Bacterial identification was carried out by 16S rRNA gene analysis and Biolog Gen Ⅲ characterization. The 16S rRNA gene sequences of 2018TS-1 and 2019SS-1 (Gene Bank: OK258319 and OK258320) isolated from T. septentrionalis and S. schlegeli were analyzed using MEGA5, and the phylogenetic tree derived from 16S rRNA gene sequences clustered the isolates with A. salmonicida. Among the Biolog Gen Ⅲ tests, 31 produced positive reactions or weak positive reactions for both strains (Dextrin, D-Maltose, D-Trehalose, D-Cellobiose, Sucrose, β-Methyl-D-Glucoside, D-Salicin, α-D-Glucose, D-Mannose, D-Fructose, D-Mannitol, Glycerol, Gelatin, Glycyl-L-Proline, L-Arginine, D-Gluconic Acid, Methyl Pyruvate, L-Malic Acid, Bromo-Succinic Acid, Tween 40, α-Keto-Butyric Acid, Acetoacetic Acid, Propionic Acid, Acetic Acid, pH 6, 1% NaCl, 1% Sodium Lactate, L-Aspartic Acid, L-Glutamic Acid, L-Histidine, and L-Serine), and two weak positive reactions for 2019SS-1, while the others were negative. According to the Biolog database, both strains were identified as A. salmonicida. Based on the molecular analysis of 16S rRNA genes and Biolog Gen Ⅲ phenotype results, the isolates were identified as A. salmonicida. The vapA gene, which encodes the outer membrane protein (A-layer protein) and causes the auto-aggregation of bacteria, is a conserved gene with some variation region in A. salmonicida. vapA gene typing is an effective and important method for classifying the molecular types and subspecies of this fish. vapA gene typing was also used in this study to identify subspecies of strains isolated from T. septentrionalis and S. schlegeli. The vapA gene sequences of 2018TS-1 and 2019SS-1 (Gene Bank: OK300094 and OK300095) were analyzed using MEGA5 with type strains obtained from Gene Bank. The phylogenetic tree derived from the vapA gene sequences clustered 2018TS-1 and 2019SS-1 with type strain ATCC 27013, indicating that the strains isolated from T. septentrionalis and S. schlegeli belonged to A. salmonicida subsp. masoucida, similar to the A-layer type Ⅶ strains, which are all from the northeast Asian and Canadian coasts in the Pacific Ocean. Based on the experimental infection, 16S rRNA sequence analysis, Biolog Gen Ⅲ characterization, and vapA gene typing, we confirmed that A. salmonicida subsp. masoucida is the pathogen of T. septentrionalis and S. schlegeli, and the cause of these two diseases on the farm. This is the first report of T. septentrionalis and S. schlegeli infected by A. salmonicida in industrial aquaculture, as well as the first report of a disease of T. septentrionalis in culture. It has been reported that A. salmonicida subsp. masoucida can infect Atlantic salmon (Salmo salar), turbot (Scophthalmus maximus), sablefish (Anoplopoma fimbria), and tongue sole (Cynoglossus semilaevis) cultured in Shandong Province. In this study, we expanded the host list of A. salmonicida subsp. masoucida to include two new species in aquaculture, T. septentrionalis and S. schlegeli, on the same farm, indicating that A. salmonicida subsp. masoucida may translate and adapt to a new host in a short period. Considering the increasing host and economic losses caused by A. salmonicida in fish culture, the prevention of A. salmonicida subsp. masoucida should be an important objective for mariculture in the future.

Background Oral squamous cell carcinoma (OSCC) is a devastating disease most often associated with tobacco consumption that induces a field of mutations from which a tumor arises. Identification of ways to prevent the emergence of cancer in high-risk patients is an ultimate goal for combatting all types of cancer, including OSCC.Methods Our study employs a mouse model of tongue carcinogenesis induced by tobacco carcinogen mimetic, 4-nitroquinoline 1-oxide (4NQO), to establish tongue dysplasia and OSCC. We use conventional histology, immunohistochemistry, multispectral imaging, mass cytometry, novel cell lines, pharmaceutical inhibition of PI3Kγ, T-cell suppression assays and mouse transplant models in our functional experimentation.Results In our study, we identify Ly6G+ granulocytes as the most abundant immune cell type in a model of tongue carcinogenesis induced by tobacco carcinogen mimetic 4NQO. Targeting Ly6G+ granulocytes with a pharmacologic inhibitor of PI3Kγ, an isoform of PI3K exclusively expressed by myeloid cells, resulted in reduced tongue dysplasia severity, and reduced rates of OSCC. Importantly, we performed functional assays with the Ly6G+ granulocytes induced in cell line models of 4NQO carcinogenesis to demonstrate that these granulocytes have increased polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) activity against T-cell proliferation and these PMN-MDSCs play a functional role in promoting tumor formation by inhibiting tumor regression in a PI3Kγ-dependent manner.Conclusions Overall, our data suggest that recruitment of PMN-MDSCs to sites of dysplasia is critical to immune suppression of CD8 T cells, thereby permitting malignancy, and PI3Kγ inhibitors are one mechanism to reduce PMN-MDSC recruitment, immunosuppression and tumorigenesis in OSCC.

Objective: Cancer stemness and M2 macrophages are intimately linked to the prognosis of lung adenocarcinoma (LUAD). For this reason, this investigation sought to identify the key genes relevant to cancer stemness and M2 macrophages, explore the relationship between these genes and clinical characteristics, and determine the potential mechanism. Methods: LUAD transcriptomic data was analyzed from The Cancer Genome Atlas (TCGA) as well as the Gene Expression Omnibus databases. Differential expression analysis was performed to discern abnormally expressed genes between LUAD and control samples in TCGA cohort. The Cell type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT) algorithm was applied to determine varyingly infiltrated immune cells in LUAD compared with the control samples in TCGA cohort. Weighted correlation network analysis (WGCNA) was performed to identify genes associated with mRNA expression-based stemness index (mRNAsi) and M2 macrophages. Least absolute shrinkage and selection operator (LASSO), RandomForest (RF) and support vector machine-recursive feature elimination (SVM-RFE) machine learning methods were conducted to detect gene signatures. Global survival evaluation (Kaplan-Meier curve) was applied to investigate the relationship between gene signatures and the survival time of LUAD patients. Receiver operating characteristic (ROC) curves were produced to define biomarkers relevant to diagnosis. Gene Set Enrichment Analysis (GSEA) was performed to probe the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to diagnostic biomarkers. The public single-cell dataset of LUAD (GSE123902) was used to investigate the expression differences of diagnostic biomarkers in various cell types in LUAD. Real-time quantitative PCR (qRT-PCR) was performed to confirm key genes in lung adenocarcinoma cells. Results: A total of 5,410 differentialy expressed genes (DEGs) as well as 15 differentially infiltrated immune cells were identified between LUAD and control sepcimens in TCGA cohort. Thirty-seven DEGs were associated with both M2 macrophages and mRNAsi according to the WGCNA analysis. Sixteen common gene signatures were obtained using three diverse algorithms. CBFA2T3, DENND3 and FCAMR were correlated to overall and disease-free survival of LUAD patients. ROC curves revealed that CBFA2T3 and DENND3 expression accurately classified LUAD and control samples. The results of single cell related analysis showed that two diagnostic biomarkers expressions were differed between the different tissue sources in M2-like macrophages. QRT-PCR demonstrated the mRNA expressions of CBFA2T3 and DENND3 were upregulated in lung adenocarcinoma cells A549 and H2122. Conclusion: Our study identified CBFA2T3 and DENND3 as key genes associated with mRNAsi and M2 macrophages in LUAD and investigated the potential molecular mechanisms underlying this relationship.

Laminin Subunit Beta 3 (LAMB3) is a transcription factor and participates in the coding of laminin. It plays an important role in cell proliferation, adhesion, and transfer by regulating various target genes and signaling pathways. However, the role of LAMB3 in human pan-cancer immunology and prognosis is still poorly understood. The TCGA, GTEx, CCLE, and HPA databases were utilized for the analysis of LAMB mRNA and protein expression. The expression of LAMB3 in various immune and molecular subtypes of human cancer was examined using the TISIDB database. The prognostic significance of LAMB3 in various cancers and clinical subtypes was investigated using Kaplan-Meier and Cox regression analysis. The relationship between LAMB3 expression, various immune cell infiltration, immune checkpoints, tumor mutational load, microsatellite instability, and DNA methylation was examined using the TCGA database. Clinical samples of four lung cancer cell lines and eight lung cancer cases were collected to confirm the expression of mRNA in lung cancer. In 17 cancers, the mRNA for LAMB3 is expressed differently and has good diagnostic and prognostic value in 22 cancers. Cox regression and Nomogram analysis show that LAMB3 is an independent risk factor for 15 cancers. LAMB3 is implicated in a variety of tumorigenesis and immune-related signaling pathways, according to GO, KEGG, and GSEA results. LAMB3 expression was associated with TMB in 33 cancer types and MSI in 32 cancer types, while in lung cancer LAMB3 expression was strongly associated with immune cell infiltration and negatively correlated with all seven methylated CpG islands. Cellular experiments demonstrated that LAMB3 promotes malignant behavior of tumor cells. Preliminary mechanistic exploration revealed its close association with PD-L1, CTLA4, cell stemness gene CD133 and β-catenin-related signaling pathways. Based on these findings, it appears that LAMB3 could be a potential therapeutic target for immunotherapy and tumor prognosis. Our findings reveal an important role for LAMB3 in different cancer types.

Objective: We are building a clinical decision support system (CSCO AI) for breast cancer patients to improve the efficiency of clinical decision-making. We aimed to assess cancer treatment regimens given by CSCO AI and different levels of clinicians. Methods: 400 breast cancer patients were screened from the CSCO database. Clinicians with similar levels were randomly assigned one of the volumes (200 cases). CSCO AI was asked to assess all cases. Three reviewers were independently asked to evaluate the regimens from clinicians and CSCO AI. Regimens were masked before evaluation. The primary outcome was the proportion of high-level conformity (HLC). Results: The overall concordance between clinicians and CSCO AI was 73.9% (3621/4900). It was 78.8% (2757/3500) in the early-stage, higher than that in the metastatic stage (61.7% [864/1400], p

The catalytic performance of metal‒organic frameworks can be tuned by introducing defects in their structure. Here, the authors introduce defects and impregnate ruthenium nanoparticles in cationic metal-organic frameworks, which enables enhanced recyclability and catalytic performance in D-glucose hydrogenation.

Abstract Background While immune checkpoint inhibitors (ICI) were approved for head and neck squamous cell carcinomas (HNSCCs), the response rate remains relatively low. Mechanisms underlying ICI unresponsiveness versus sensitivity are not fully understood. Method To better delineate differential responses to ICI treatment, we employed mouse SCC models, termed KPPA tumors that were caused by deleting p53 and hyperactivating PIK3CA, two most frequently mutated genes in human HNSCCs. We transplanted two KPPA tumor lines (TAb2 versus TCh3) into C57BL/6 recipients and examined the immune tumor microenvironment using flow cytometry. Furthermore, we employed single-cell RNA sequencing to identify the difference in tumor infiltrating lymphocytes (TILs). Results We found that different KPPA tumors exhibited heterogeneous immune profiles pre-existing treatment that dictated their sensitivity or unresponsiveness to anti-PD-L1. Unresponsive TAb2 tumors were highly enriched with functional tumor-associated macrophages (TAMs), especially M2-TAMs. In contrast, sensitive TCh3 tumors contained more CD8 TILs with better effector functions. TAb2 tumor cells drastically expanded F4/80+ TAMs from bone marrow precursors, requiring CSF1 and VEGF. Consistently, a higher combined expression of VEGF-C and CSF1 predicts worse survival in PIK3CAAmp/TP53Mutated HNSCC patients. Unresponsive TAb2 tumors upregulated distinct signaling pathways that correlate with aggressive tumor phenotypes. While anti-PD-L1 did not affect the TME of TAb2 tumors, it significantly increased the number of CD8 TILs in TCh3 tumors. Conclusions We uncovered tumor-intrinsic differences that may underlie the differential responses to ICI by establishing and employing two SCC tumor lines, TAb2 vs. TCh3, both of which harbor TP53 deletion and PIK3CA hyperactivation. Our study indicates the limitation of stratifying cancers according to their genetic alterations and suggests that evaluating HNSCC tumor-intrinsic cues along with immune profiles in the TME may help better predict ICI responses. Our experimental models may provide a platform for pinpointing tumor-intrinsic differences underlying an immunosuppressive TME in HNSCCs and for testing combined immunotherapies targeting either tumor-specific or TAM-specific players to improve ICI efficacy.

Background and objective The previous study has indicated that lung cancer has a high incidence and mortality in China, and has caused a large economic burden. The purpose of this study was to analyze the incidence and economic burden of lung cancer by analyzing the information on the home page of discharge history of lung cancer patients in Hebei Tumor Hospital, and to provide scientific basis for the prevention and treatment of lung cancer. Methods The information of all of the discharges, new cases, surgical patients, age, gender, length of stay and hospitalization cost of lung cancer patients in Hebei Tumor Hospital from January 2012 to December 2019 were retrieved based on the medical record management system, and the incidence trend, gender and age distribution as well as the economic burden of the disease were statistically described. Results The number of new cases of lung cancer increased year by year, from 2,235 cases in 2012 to 5,012 cases in 2019. The number of males always outnumbered females, but the gender ratio decreased year by year, from 2.25 in 2012 to 1.56 in 2019. Among new cases of lung cancer, the proportion of surgical treatment increased year by year, from 28.14% in 2012 to 44.83% in 2019. Except for 2012, the proportion of surgical operations in female patients was higher than that in male patients from 2013 to 2019. The proportion of surgical operations in male and female patients was 23.52% and 28.07% in 2013, and 36.14% and 58.37% in 2019, respectively. The median age at the onset of lung cancer has increased year by year, from 61 years old in 2012 to 63 years old in 2019. The median age of onset in all lung cancer patients was higher in males than in females. The number of new lung cancer patients and surgical patients both showed an increasing trend with the increase of age, and both reached the maximum value in the age group of 60-69 years old. With the increase of age, the number of patients gradually decreased. The median length of hospital stay for all discharged lung cancer patients or surgical patients decreased year by year, from 10 d and 19 d in 2012 to 8 d and 17 d in 2019, respectively, while the median hospitalization cost increased year by year. It increased from 10,611.46 yuan and 38,750.13 yuan in 2012 to 17,187.15 yuan and 84,030.16 yuan in 2019, respectively. Conclusion Lung cancer is still one of the main cancers endangering the health of Chinese residents. The incidence of lung cancer is increasing year by year, and the distribution of gender and age has certain characteristics. In order to reduce the number of cases and the economic burden, effective prevention and control measures should be formulated and medical reform should be strengthened.

Macrophages are one of the most important players in the tumor microenvironment. But the contribution of macrophages to lung adenocarcinoma (LUAD) is still controversial. The current study aimed to display an immune landscape to clarify the function of macrophages and detect prognostic hub genes in LUAD. The transcriptome data were adopted to screen differently expressed genes (DEGs) in The Cancer Genome Atlas database (TCGA). The cell type identification by estimating relative subsets of RNA transcripts (CIBERSORT) algorithm was used to reveal the immune landscape. Weighted gene co-expression network analysis (WGCNA) analysis was performed to identify the hub module associated with macrophages. Function Enrichment analysis was conducted on hub module genes. Moreover, univariate and multivariate Cox regression analyses were performed to identify prognostic hub genes. Kaplan-Meier (KM) and Time-dependent receiver operating characteristic (ROC) curves were plotted to assess the prognostic capacity of the four prognostic hub genes. The GES1196959 dataset from the Gene Expression Omnibus (GEO) database was downloaded to verify the differential expression of the 4 prognostic hub genes.

BackgroundContinuous theta burst stimulation (cTBS) is a specific paradigm of repetitive transcranial magnetic stimulation (rTMS) with an inhibitory effect on cortical excitability for up to 60 min after less than 1 min of stimulation. The right posterior superior temporal gyrus (pSTG), homotopic to Wernicke’s area in the left hemisphere, may be a potential stimulation target based on its critical role in semantic processing. The objective of this study was to explore whether cTBS over the right pSTG can promote language improvements in aphasic patients and the underlying mechanism.MethodsA total of 34 subjects with aphasia were randomly assigned to undergo 15 sessions of either 40-s inhibitory cTBS over the right pSTG (the cTBS group) or sham stimulation (the sham group), followed by 30 min of speech and language therapy. Subjects underwent resting-state functional magnetic resonance imaging (rs-fMRI), and the aphasia quotient (AQ) of the Chinese version of the Western Aphasia Battery (WAB) was calculated before and after the intervention. This randomized controlled trial was registered in the Chinese Clinical Trial Registry (No. ChiCTR210052962).ResultsAfter treatment, the language performance of the cTBS group was higher than that of the sham group in terms of the WAB-AQ score (p = 0.010) and the WAB scores for auditory comprehension (p = 0.022) and repetition (p = 0.035). The fractional amplitude of low-frequency fluctuations (fALFF) was significantly decreased in the pars triangularis of the inferior frontal gyrus (IFG), right middle frontal gyrus, right thalamus, and left cerebellar crus I. Clusters in the left orbitofrontal cortex exhibited increased fALFF. The change in WAB comprehension scores were significantly correlated with the change in the fALFF of the right IFG pars triangularis in both groups. Greatly increased functional connectivity was observed between the right pars triangularis and left paracingulate gyrus and between the right pSTG and right angular gyrus and the posterior cingulate gyrus with pre-and post-treatment between the two groups.ConclusionOur findings indicate that cTBS of the right pSTG may improve language production by suppressing intrinsic activity of the right fronto-thalamic-cerebellar circuit and enhancing the involvement of the right temporoparietal region.

Offshore cephalopods usually experience a short and extreme senescence process at the end of their lives. Unique behaviors such as brooding and fasting occur during this period. The cause of senescent cephalopod death is still unknown. Excessive energy consumption under prolonged starvation is one possible reason for death. A comparison analysis between normal starvation and senescence may provide valuable information to deepen our understanding of the senescence process of cephalopods. In this study, a comparison of behavior, histology and expression of a candidate senescence-specific gene (Imaginal morphogenesis protein-Late 2, ImpL2) of female Octopus sinensis under normal starvation and senescence was conducted. The females in the starvation group had similar behaviors, histology and ImpL2 expression level to the control group. The females in the senescence group exhibited typical behaviors and symptoms of senescent octopus, including fasting and unhealed lesions on the skin. Moreover, swimming and grasping ability was weakened in senescent individuals. The muscle tissue in the mantle and arm of senescent females showed a histolytic appearance. The secretory cells and their microvilli in the digestive gland gradually degraded and disappeared during senescence. The expression level of ImpL2 gene significantly changes in multiple organs of senescent females. The results showed that O. sinensis underwent different physiological and molecular processes during starvation and senescence. Starvation alone could not lead to senescent symptoms in the octopus. Tissue degeneration in the mantle, arm and digestive gland would increase the death risk of the octopus.

The complete mitochondrial genome of the East Asian common octopus (Octopus sinensis) was determined and analyzed in this work. The circular mitogenome of O. sinensis is 15,737 bp in length with 21.53% GC content, which contains two ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs), 13 protein-coding genes (PCGs) and a non-coding region (D-loop). The analysis of the SNPs revealed 4 conservative SNPs for COI gene of O. sinensis compared with Octopus vulgaris. Phylogenetic analysis suggested that O. sinensis is closely related to O. vulgaris. This sequence data would play an important role in the investigation of phylogenetic relationship and taxonomy of the class Cephalopoda.

Abstract. Increasing numbers of targeted drugs are used in hormone receptor (HR)-positive metastatic breast cancer (MBC) to overcome or delay resistance to endocrine therapy. This study will systemically review the progress made in endocrine therapy combined with targeted therapy in the treatment of HR-positive MBC. From the “AI (aromatase inhibitor) era” represented by aromatase inhibitors, we have gradually entered the “post-AI era” represented by fulvestrant. Under the guidance of research on the molecular mechanism of endocrine therapy resistance, the “combination of endocrine therapy and targeted therapy” era is approaching. The development of drugs that target endocrine therapy resistance has concentrated on cyclin-dependent kinase 4/6 inhibitors, histone deacetylase inhibitors, and inhibitors of drug targets in the phosphatidylinositol 3 kinase-protein kinase B-mammalian target of rapamycin (PI3K-AKT-mTOR) pathway, providing new strategies for HR-positive MBC. Exploring biomarkers to guide the more precise use of targeted drugs in endocrine therapy for MBC is the focus of current and future research.

Objective: Lung adenocarcinoma (LUAD) is a common malignant tumor with a poor prognosis. The present study aimed to screen the key genes involved in LUAD development and prognosis. Methods: The transcriptome data for 515 LUAD and 347 normal samples were downloaded from The Cancer Genome Atlas and Genotype Tissue Expression databases. The weighted gene co-expression network and differentially expressed genes were used to identify the central regulatory genes for the development of LUAD. Univariate Cox, LASSO, and multivariate Cox regression analyses were utilized to identify prognosis-related genes. Results: The top 10 central regulatory genes of LUAD included IL6, PECAM1, CDH5, VWF, THBS1, CAV1, TEK , HGF, SPP1, and ENG . Genes that have an impact on survival included PECAM1, HGF, SPP1, and ENG . The favorable prognosis genes included KDF1, ZNF691, DNASE2B, and ELAPOR1, while unfavorable prognosis genes included RPL22, ENO1, PCSK9, SNX7, and LCE5A. The areas under the receiver operating characteristic curves of the risk score model in the training and testing datasets were .78 and .758, respectively. Conclusion: Bioinformatics methods were used to identify genes involved in the development and prognosis of LUAD, which will provide a basis for further research on the treatment and prognosis of LUAD.

Despite advances in modern control theory and artificial intelligence technology, current methods for tuning proportional-integral-derivative (PID) controller parameters based on the traditional particle swarm optimization (PSO) algorithm do not meet the requirements for controlling an unmanned surface vessel (USV) propulsion motor. To overcome the disadvantages of the PSO algorithm, such as low precision and easily falling into a local optimum, the beetle antennae search (BAS) algorithm can be introduced into the PSO algorithm by replacing particles with beetles, and effectively prevents the PSO algorithm from easily falling into the local optimum. At the same time, the BAS algorithm will no longer be limited to single objective parameterization. Herein, we propose a PID parameter optimization method based on the hybrid BAS-PSO algorithm for a USV propulsion motor. The PID parameter optimization of propulsion motor effectively becomes a beetle foraging problem with group optimization. Numerical results show that the method can effectively solve the problems of PSO and greatly improve convergence speed. Compared with the genetic algorithm and standard PSO algorithm, the BAS-PSO algorithm is superior for PID parameter tuning and can improve performance of USV propulsion system.

Some standards can no longer meet the requirements of the development of the situation,since they have been used for quite a long time.The enterprise standard is the revision and improvement of the existing standard system,which can effectively make up for the shortcomings of the current standards.Therefore,based on national standard of wheat flour,combined with the document of the special wheat flour standard of Guangdong province,the problems existing in the enterprise standard of special wheat flour,such as incorrect expression and text irrational physical and chemical indicators,wrong normative reference,lack of product quality indicators,and the relatively simple standard category of the special wheat flour for the enterprise filing,are discussed.Some suggestions are proposed to provide some valuable references for the revision of enterprise standard.

In this study, a new method was developed for the comprehensive quality evaluation (QE) of Artemisia capillaris Thunb. (A. capillaris, named Yinchenhao in Chinese), which is one of the most commonly used herbal medicines (HMs). First, fingerprints of 31 batch samples of A. capillaris were determined by HPLC, the reference fingerprint was established, and the common peaks were assigned. Second, the components of common peaks in the HPLC fingerprints were identified by ultrafast liquid chromatography- (UFLC-) Q-TOF-MS/MS. Finally, the contents of the components unambiguously confirmed by reference substances were determined, and the correlation between the contents of chlorogenic acid and the contents of others was analyzed. The results showed that there were 20 common peaks in the HPLC fingerprints of 31 batch samples. The components of these 20 common peaks were identified as ten organic acids, eight flavonoids, and two others. Among nine organic acids such as 1-caffeoylquinic acid, neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid, three flavonoids such as rutin, hyperoside, and isoquercetin, and one other p-hydroxyacetophenone, a total of 13 ones were unambiguously identified by comparison with reference substances; one caffeoylquinic acid glucoside and one flavone di-C-glucoside were detected in A. capillaris for the first time. There were some differences in the contents of 13 components in different samples; chlorogenic acid could be regarded as the quality marker of A. capillaris. The current established method in this study can be used for the comprehensive QE of A. capillaris and can also provide reference for the QE of the other HMs.

Background: The addition of anti-human epidermal growth factor receptor 2 (HER2)-targeted drugs, such as trastuzumab, lapatinib, and trastuzumab emtansine (T-DM1), to chemotherapy significantly improved prognosis of HER2-positive breast cancer patients. However, it was confused that metastatic patients vary in the response of targeted drug. Therefore, methods of accurately predicting drug response were really needed. To overcome the spatial and temporal limitations of biopsies, we aimed to develop a more sensitive and less invasive method of detecting mutations associated with anti-HER2 therapeutic response through circulating-free DNA (cfDNA). Methods: From March 6, 2014 to December 10, 2014, 24 plasma samples from 20 patients with HER2-positive metastatic breast cancer who received systemic therapy were eligible. We used a panel for detection of hot-spot mutations from 50 oncogenes and tumor suppressor genes, and then used targeted next-generation sequencing (NGS) to identify somatic mutation of these samples in those 50 genes. Samples taken before their first trastuzumab administration and subsequently proven with clinical benefit were grouped into sensitive group. The others were collected after disease progression of the trastuzumab-based therapy and were grouped into the resistant group. Results: A total of 486 single-nucleotide variants from 46 genes were detected. Of these 46 genes, phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), proto-oncogene c-Kit (KIT), and tumor protein p53 (TP53) were the most common mutated genes. Seven genes, including epidermal growth factor receptor (EGFR), G protein subunit alpha S (GNAS), HRas proto-oncogene (HRAS), mutL homolog 1 (MLH1), cadherin 1 (CDH1), neuroblastoma RAS viral oncogene homolog (NRAS), and NOTCH1, that only occurred m utations in the resistant group were associated with the resistance of targeted therapy. In addition, we detected a HER2 S855I mutation in two patients who had persistent benefits from anti-HER2 therapy. Conclusion: Targeted NGS of cfDNA has potential clinical utility to detect biomarkers from HER2-targeted therapies.

The first complete mitochondrial genome of the wolf-eel (Anarrhichthys ocellatus) was determined and analyzed in this work. It had a circular mapping molecular with the length of 16,506 bp and contained 12S and 16S rRNAs, 22 tRNAs, 13 protein-coding genes, and a 851 bp D-loop in the typical arrangement of the vertebrate consensus. Phylogenetic analysis of the mitochondrial genome sequences of 47 representative species within the order Perciformes suggests that A. ocellatus is closely related to the species in the family Anarhichadidae. Anarrhichthys ocellatus mitogenome can contribute to our understanding of the phylogeny and evolution of this species.

Automatic detection system has been widely used in pavement detection. This paper describes the principle of shooting width adjustment and method of adjusting the width of the image acquisition. This system increased width of pavement video acquisition system from maximum width of 5 meters at present to full face pavement 12 meters.It is proved through testing sections that it achieved good image effect and met the design requirements.

Understanding how genetic variants impact molecular phenotypes is a key goal of functional genomics, currently hindered by reliance on a single haploid reference genome. Here, we present the EN-TEx resource of 1,635 open-access datasets from four donors (∼30 tissues × ∼15 assays). The datasets are mapped to matched, diploid genomes with long-read phasing and structural variants, instantiating a catalog of >1 million allele-specific loci. These loci exhibit coordinated activity along haplotypes and are less conserved than corresponding, non-allele-specific ones. Surprisingly, a deep-learning transformer model can predict the allele-specific activity based only on local nucleotide-sequence context, highlighting the importance of transcription-factor-binding motifs particularly sensitive to variants. Furthermore, combining EN-TEx with existing genome annotations reveals strong associations between allele-specific and GWAS loci. It also enables models for transferring known eQTLs to difficult-to-profile tissues (e.g., from skin to heart). Overall, EN-TEx provides rich data and generalizable models for more accurate personal functional genomics.

Fms-like tyrosine kinase 3 ligand (Flt3L) is known as the primary differentiation and survival factor for dendritic cells (DCs). Furthermore, Flt3L is involved in the homeostatic feedback loop between DCs and regulatory T cell (Treg). We have previously shown that Flt3L accumulates in the synovial fluid in rheumatoid arthritis (RA) and that local exposure to Flt3L aggravates arthritis in mice, suggesting a possible involvement in RA pathogenesis. In the present study we investigated the role of Flt3L on DC populations, Tregs as well as inflammatory responses in experimental antigen-induced arthritis. Arthritis was induced in mBSA-immunized mice by local knee injection of mBSA and Flt3L was provided by daily intraperitoneal injections. Flow cytometry analysis of spleen and lymph nodes revealed an increased formation of DCs and subsequently Tregs in mice treated with Flt3L. Flt3L-treatment was also associated with a reduced production of mBSA specific antibodies and reduced levels of the pro-inflammatory cytokines IL-6 and TNF-α. Morphological evaluation of mBSA injected joints revealed reduced joint destruction in Flt3L treated mice. The role of DCs in mBSA arthritis was further challenged in an adoptive transfer experiment. Transfer of DCs in combination with T-cells from mBSA immunized mice, predisposed naïve recipients for arthritis and production of mBSA specific antibodies. We provide experimental evidence that Flt3L has potent immunoregulatory properties. Flt3L facilitates formation of Treg cells and by this mechanism reduces severity of antigen-induced arthritis in mice. We suggest that high systemic levels of Flt3L have potential to modulate autoreactivity and autoimmunity.

BackgroundTwo major forms of inherited J-wave syndrome (JWS) are recognized: early repolarization syndrome (ERS) and Brugada syndrome (BrS).ObjectivesThis study sought to assess the distinct features between patients with ERS and BrS carrying pathogenic variants in SCN5A.MethodsClinical evaluation and next-generation sequencing were performed in 262 probands with BrS and 104 with ERS. Nav1.5 and Kv4.3 channels were studied with the use of patch-clamp techniques. A computational model was used to investigate the protein structure.ResultsThe SCN5A+ yield in ERS was significantly lower than in BrS (9.62% vs 22.90%; P = 0.004). Patients diagnosed with ERS displayed shorter QRS and QTc than patients with BrS. More than 2 pathogenic SCN5A variants were found in 5 probands. These patients displayed longer PR intervals and QRS duration and experienced more major arrhythmia events (MAE) compared with those carrying only a single pathogenic variant. SCN5A-L1412F, detected in a fever-induced ERS patient, led to total loss of function, destabilized the Nav1.5 structure, and showed a dominant-negative effect, which was accentuated during a febrile state. ERS-related SCN5A-G452C did not alter the inward sodium current (INa) when SCN5A was expressed alone, but when coexpressed with KCND3 it reduced peak INa by 44.52% and increased the transient outward potassium current (Ito) by 106.81%.ConclusionsThese findings point to SCN5A as a major susceptibility gene in ERS as much as it is in BrS, whereas the lower SCN5A+ ratio in ERS indicates the difference in underlying electrophysiology. These findings also identify the first case of fever-induced ERS and demonstrate a critical role of Ito in JWS and a higher risk for MAE in JWS probands carrying multiple pathogenic variants in SCN5A.