An immune cell atlas reveals the dynamics of human macrophage specification during prenatal development.

Macrophages are heterogeneous and play critical roles in development and disease, but their diversity, function, and specification remain inadequately understood during human development. We generated a single-cell RNA sequencing map of the dynamics of human macrophage specification from PCW 4-26 across 19 tissues. We identified a microglia-like population and a proangiogenic population in 15 macrophage subtypes. Microglia-like cells, molecularly and morphologically similar to microglia in the CNS, are present in the fetal epidermis, testicle, and heart. They are the major immune population in the early epidermis, exhibit a polarized distribution along the dorsal-lateral-ventral axis, and interact with neural crest cells, modulating their differentiation along the melanocyte lineage. Through spatial and differentiation trajectory analysis, we also showed that proangiogenic macrophages are perivascular across fetal organs and likely yolk-sac-derived as microglia. Our study provides a comprehensive map of the heterogeneity and developmental dynamics of human macrophages and unravels their diverse functions during development.

Biological functions of miRNA-203b-3p/ZNF324 in laryngeal carcinoma.

The present study aimed to elucidate the role of MicroRNA-203b-3p (miRNA-203b-3p) in protecting the deterioration of laryngeal carcinoma through targeting ZNF324. Relative levels of miRNA-203b-3p and ZNF324 in laryngeal carcinoma tissues with different tumor node metastasis (TNM) staging and pathological grades were detected. Regulatory effects of miRNA-203b-3p on clonality, viability and 5-Ethynyl-2'- deoxyuridine (EdU)-positive ratio in M2E and TU212 cells were assessed. The binding relationship between miRNA-203b-3p and ZNF324 was evaluated by dual-luciferase reporter gene assay. The involvement of ZNF324 in cell phenotype changes of laryngeal carcinoma regulated by miRNA-203b-3p was explored by rescue experiments. MiRNA-203b-3p was downregulated in laryngeal carcinoma, especially in those with advanced TNM staging or pathological grade. Overexpression of miRNA-203b-3p reduced clonality, viability and EdU-positive ratio in M2E and TU212 cells. In addition, ZNF324 was upregulated in laryngeal carcinoma, which was negatively regulated by miRNA-203b-3p. ZNF324 was verified to be the target binding miRNA-203b-3p. Notably, overexpression of ZNF324 could partially reverse the inhibitory effects of miRNA-203b-3p on laryngeal carcinoma proliferation. MiRNA-203b-3p is downregulated in laryngeal carcinoma, which blocks laryngeal carcinoma cells to proliferate through targeting ZNF324 and thus alleviates cancer progression.

A Questionnaire-Based Ensemble Learning Model to Predict the Diagnosis of Vertigo: Model Development and Validation Study.

BACKGROUND: Questionnaires have been used in the past 2 decades to predict the diagnosis of vertigo and assist clinical decision-making. A questionnaire-based machine learning model is expected to improve the efficiency of diagnosis of vestibular disorders. OBJECTIVE: This study aims to develop and validate a questionnaire-based machine learning model that predicts the diagnosis of vertigo. METHODS: In this multicenter prospective study, patients presenting with vertigo entered a consecutive cohort at their first visit to the ENT and vertigo clinics of 7 tertiary referral centers from August 2019 to March 2021, with a follow-up period of 2 months. All participants completed a diagnostic questionnaire after eligibility screening. Patients who received only 1 final diagnosis by their treating specialists for their primary complaint were included in model development and validation. The data of patients enrolled before February 1, 2021 were used for modeling and cross-validation, while patients enrolled afterward entered external validation. RESULTS: A total of 1693 patients were enrolled, with a response rate of 96.2% (1693/1760). The median age was 51 (IQR 38-61) years, with 991 (58.5%) females; 1041 (61.5%) patients received the final diagnosis during the study period. Among them, 928 (54.8%) patients were included in model development and validation, and 113 (6.7%) patients who enrolled later were used as a test set for external validation. They were classified into 5 diagnostic categories. We compared 9 candidate machine learning methods, and the recalibrated model of light gradient boosting machine achieved the best performance, with an area under the curve of 0.937 (95% CI 0.917-0.962) in cross-validation and 0.954 (95% CI 0.944-0.967) in external validation. CONCLUSIONS: The questionnaire-based light gradient boosting machine was able to predict common vestibular disorders and assist decision-making in ENT and vertigo clinics. Further studies with a larger sample size and the participation of neurologists will help assess the generalization and robustness of this machine learning method.

miR-34a Suppresses Cell Proliferation in Laryngeal Cancer by Targeting Prominin 1.

The study explored the cross-talk between the microRNA miR-34a and prominin 1 in the development of laryngeal cancer (LC), which has an unacceptable high mortality rate. We predicted that miR-34a might target prominin 1. miR-34a and prominin 1 expression was analyzed with reverse transcription quantitative polymerase chain reaction. The correlation between miR-34a and prominin 1 was determined by linear regression analysis. miR-34a and prominin 1 overexpression and miR-34a inhibition were achieved in LC cells to analyze their relationship. The roles of miR-34a and prominin 1 in LC cell proliferation were studied with CCK-8 assay. The results showed miR-34a was downregulated, and prominin 1 was upregulated in LC. In addition, prominin 1 and miR-34a were inversely correlated across cancer tissue samples. In cancer cells, miR-34a overexpression downregulated prominin 1, while miR-34a knockdown upregulated prominin 1. Moreover, miR-34a overexpression reduced the enchaining effects of prominin 1 on LC cell proliferation. Therefore, miR-34a might suppress LC cell proliferation by targeting prominin 1.

Intranasal fluticasone furoate in pediatric allergic rhinitis: randomized controlled study.

BACKGROUND: Intranasal corticosteroids are the most efficacious anti-inflammatory medications for allergic rhinitis (AR). However, the efficacy and safety of intranasal corticosteroids in children have not yet been subject to specific research in China. The aim of this study was to investigate the efficacy and safety of fluticasone furoate nasal spray (FFNS) in a Chinese pediatric population. METHODS: In this phase 4 randomized, double-blind, placebo-controlled, multicenter study, pediatric AR patients aged 2-12 years were randomized 1:1:1, receiving either FFNS 55 µg or 110 µg or placebo. Electronic diary cards were completed to record symptoms, rescue medication use, and treatment compliance. Anterior rhinoscopy and overall response to therapy were evaluated and recorded. RESULTS: Patients treated with FFNS at either dose experienced a significantly greater reduction in daily reflective total nasal symptom score compared with placebo. This was maintained in a younger subset of patients (2-6 years). Drug-related adverse events occurred in <20% of patients in all groups. FFNS was well tolerated at both doses. CONCLUSIONS: This study demonstrates favorable efficacy and safety profiles for FFNS 55 µg or 110 µg in Chinese pediatric populations (2-12 years), supporting its use in clinical treatment for AR children, including younger children aged 2-6 years. IMPACT: The aim of this study was to investigate the efficacy and safety of intranasal fluticasone furoate in Chinese pediatric allergic rhinitis. This research not only addresses the deficiency in efficacy and safety data for intranasal corticosteroids in very young patients (aged 2-6 years) worldwide but also demonstrates that fluticasone furoate nasal spray shows a favorable benefit/risk profile at different dose levels. Our data will be of interest to the broad readership of Pediatric Research and will positively contribute to the dialog regarding the treatment of allergic rhinitis in children aged 2-6 years.

An epidemiological survey of laryngopharyngeal reflux disease at the otorhinolaryngology-head and neck surgery clinics in China.

PURPOSE: Using the Reflux Symptom Index (RSI), this nationwide study aimed to investigate the incidence, diagnostic status, risk factors, and common symptoms of adult laryngopharyngeal reflux disease (LPRD) at otorhinolaryngology-head and neck surgery (OHNS) clinics in China. METHODS: This multicenter cross-sectional survey began at the different institutions ranged from July to October 2017, and the duration was 12 months. A total of 90,440 eligible patients were finally enrolled from 72 medical institutions in China. All these patients completed the questionnaire based on RSI. In this study, LPRD was defined as RSI > 13. RESULTS: There were 9182 with LPRD among the 90,440 eligible participants (10.15%). However, only 1294 had a history of LPRD diagnosis among those with LPRD (14.09%). There were regional differences in the frequency of LPRD (P < 0.001). The proportions of patients with LPRD in males (vs. females), middle- and old-aged patients (vs. young), with current smoking history (vs. no smoking), and current drinking history (vs. no drinking) were significantly higher (all P < 0.001). Middle and old age, current smoking, and drinking history were independent predictors of LPRD (all P < 0.001, OR 1.240, 1.261, and 1.481, respectively). "Sensations of something stuck in throat or a lump in throat", "clearing throat", and "excess throat mucus or postnasal drip" were the most frequent clinical symptoms in patients with LPRD. CONCLUSIONS: LPRD has a high incidence at the OHNS clinics in China. However, the diagnostic status of this disease is not optimistic. Older age, smoking, and drinking history were risk factors for LPRD.

Correction to: Surface-enhanced Raman spectroscopy of blood serum based on gold nanoparticles for the diagnosis of the oral squamous cell carcinoma.

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Surface-enhanced Raman spectroscopy of blood serum based on gold nanoparticles for the diagnosis of the oral squamous cell carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is becoming more common across the globe. The prognosis of OSCC is largely dependent on the early detection. But the routine oral cavity examination may delay the diagnosis because the early oral malignant lesions may be clinically indistinguishable from benign or inflammatory diseases. In this study, the new diagnostic method is developed by using the surface enhanced Raman spectroscopy (SERS) to detect the serum samples from the cancer patients. METHOD: The blood serum samples were collected from the OSCC patients, MEC patients and the volunteers without OSCC or MEC. Gold nanoparticles(NPs) were then mixed in the serum samples to obtain the high quality SERS spectra. There were totally 135 spectra of OSCC, 90 spectra of mucoepidermoid carcinoma (MEC) and 145 spectra of normal control group, which were captured by SERS successfully. Compared with the normal control group, the Raman spectral differences exhibited in the spectra of OSCC and MEC groups, which were assigned to the nucleic acids, proteins and lipids. Based on these spectral differences and features, the algorithms of principal component analysis(PCA) and linear discriminant analysis (LDA) were employed to analyze and classify the Raman spectra of different groups. RESULTS: Compared with the normal groups, the major increased peaks in the OSCC and MEC groups were assigned to the molecular structures of the nucleic acids and proteins. And these different major peaks between the OSCC and MEC groups were assigned to the special molecular structures of the carotenoids and lipids. The PCA-LDA results demonstrated that OSCC could be discriminated successfully from the normal control groups with a sensitivity of 80.7% and a specificity of 84.1%. The process of the cross validation proved the results analyzed by PCA-LDA were reliable. CONCLUSION: The gold NPs were appropriate substances to capture the high-quality SERS spectra of the OSCC, MEC and normal serum samples. The results of this study confirm that SERS combined PCA-LDA had a giant capability to detect and diagnosis OSCC through the serum sample successfully.

[Airborne fine particle decreases the cell viability and induces inflammation in human bronchial epithelial cells].

OBJECTIVE: To investigate the effects of airborne fine particle on cell viability and inflammation in human bronchial epithelial cells.
 Methods: Atmospheric PM2.5 samples were collected by PM2.5 sampler. PM2.5 morphology was observed by scanning electron microscope (SEM). Human bronchial epithelial cells (BEAS-2B) were treated with PM2.5 at different concentrations (0, 50, 100, 200, 400, 800 µg/mL) for 12, 24 or 48 hours, and the cell activity were evaluated by cell counting kit-8 (CCK-8). The mRNA expression levels of (granulocyte-macrophage colony stimulating factor，GM-CSF) and TNF-α were detected by quantitative real-time PCR (qRT-PCR). Western blot was used to detect the protein expressions of GM-CSF and TNF-α.
 Results: According to SEM, the shape of PM2.5 varied, and the diameter was different and mostly equal to or less than 2.5 µm. CCK-8 assay showed that different concentrations of PM2.5 exposure for 12 hours, 24 hours and 48 hours resulted in loss of cell viability of BEAS-2B cells (P<0.05). Different concentrations of PM2.5 increased the mRNA and protein expression of GM-CSF and TNF-α, and the higher concentration of PM2.5 induced higher expression, which have statistical significant difference between the groups (P<0.05).
 Conclusion: Atmospheric PM2.5 can cause inflammatory response in human bronchial epithelial cells. They can reduce cell viability, which may be related to the PM2.5 trigger and aggravation of bronchopulmonary inflammatory diseases.

Label-free blood serum detection by using surface-enhanced Raman spectroscopy and support vector machine for the preoperative diagnosis of parotid gland tumors.

BACKGROUND: It is difficult for the parotid gland neoplasms to make an accurate preoperative diagnosis due to the restriction of biopsy in the parotid gland neoplasms. The aim of this study is to apply the surface-enhanced Raman spectroscopy (SERS) method for the blood serum biochemical detection and use the support vector machine for the analysis in order to develop a simple but accurate blood serum detection for preoperative diagnosis of the parotid gland neoplasms. METHODS: The blood serums were collected from four groups: the patients with pleomorphic adenoma, the patients with Warthin's tumor, the patients with mucoepidermoid carcinoma and the volunteers without parotid gland neoplasms. Au nanoparticles (Au NPs) were mixed with the blood serum as the SERS active nanosensor to enhance the Raman scattering signals produced by the various biochemical materials and high quality SERS spectrum were obtained by using the Raman microscope system. Then the support vector machine was utilized to analyze the differences of the SERS spectrum from the blood serum of different groups and established a diagnostic model to discriminate the different groups. RESULTS: It was demonstrated that there were different intensities of SERS peaks assigned to various biochemical changes in the blood serum between the parotid gland tumor groups and normal control group. Compared with the SERS spectra of the normal serums, the intensities of peaks assigned to nucleic acids and proteins increased in the SERS spectra of the parotid gland tumor serums, which manifested the differences of the biochemical metabolites in the serum from the patients with parotid gland tumors. When the leave-one-sample-out method was used, support vector machine (SVM) played an outstanding performance in the classification of the SERS spectra with the high accuracy (84.1 % ~ 88.3 %), sensitivity (82.2 % ~ 97.4 %) and specificity (73.7 % ~ 86.7 %). Though the accuracy, sensitivity and specificity decreased in the leave-one-patient-out cross validation, the mucoepidermoid carcinoma was still easier to diagnose than other tumors. DISCUSSION: The specific molecular differences of parotid gland tumors and normal serums were significantly demonstrated through the comparison between the various SERS spectra.But compared with the serum SERS spectra reported in the other studies, some differences exist between the spectra in this study and the ones reported in the lietratures. These differences may result from the various nano-particles, the different preparation of serum and equipment parameters, and we could need a further research to find an exact explanation.Based on the SERS spectra of the serum samples, SVM have shown a giant potential to diagnose the parotid gland tumors in our preliminary study. However, different cross validaiton methods could effect the accuracy and a further study involing a great number of samples should be needed. CONCLUSIONS: This exploratory research demonstrated the great potential of SERS combined with SVM into a non-invasive clinical diagnostic method for preoperative diagnosis of parotid gland tumors. And the internal relation between the spectra and patients should be established in the further study.

Neoadjuvant therapy with chemotherapy and immune checkpoint inhibitor for laryngeal function preservation in locally advanced hypopharyngeal cancer.

Purpose: To evaluate the efficacy and laryngeal function preservation of neoadjuvant treatment with chemotherapy and immune checkpoint inhibitor for locally advanced hypopharyngeal cancer (LAHPC). Methods: We retrospectively collected LAHPC patients who were diagnosed between February 2022 and June 2023. The patients received a combination of chemotherapy and immune checkpoint inhibitors as the neoadjuvant therapy. The response to treatment, laryngeal function preservation rate, and short-term survival were assessed. Results: A total of 20 patients were included. Of these patients, 17 (85.0%) had stage IVA-B disease. Ten (50%) and four (20%) patients achieved pathological complete response (PCR) and major pathological response (MPR) to the primary tumor, respectively. In addition, 6 patients had incomplete pathological response (IPR). In the neck, 19 patients had node-positive disease before treatment, and only 5 patients (26.4%) had PCR to regional lymph nodes. Pathologically positive lymph nodes were still observed in 14 (73.6%) patients. Significant downgrading on narrow-band imaging assessment in primary tumors was associated with a higher probability of PCR or MPR than those with IPR (92.9% vs. 33.3%, P=0.014). The overall rate of laryngeal preservation was 95.0%. No severe perioperative complications or perioperative death were found. All patients completed the recommended postoperative radiotherapy/chemoradiotherapy. The median follow-up period was 12.1 months. The 1-year progression-free survival and overall survival were 94.1% and 92.9%, respectively. During the follow-up period, all 19 patients who underwent laryngeal preservation surgery had their laryngeal function preserved. Conclusion: The addition of an immune checkpoint inhibitor to neoadjuvant chemotherapy effectively preserves laryngeal function without increasing complications related to surgery and postoperative radiotherapy in LAHPC.

Preliminary study on the resection of parapharyngeal and lateral skull base tumors by using transoral endoscopy with 3D visualization and navigation technologies.

OBJECTIVES: With the assistance of 3D visualization and real-time navigation technologies, the tumors in the parapharyngeal and lateral skull base should be removed through oral the approach with endoscopy. METHODS: The preoperative CT data of eight patients with parapharyngeal or lateral skull base soft tissue tumors were modeled, and the anatomical position relationship between the tumor and surrounding blood vessels and other important structures was reconstructed using 3D visualization technology, and preoperative design was performed. The intraoperative oral approach and real-time navigation guidance were adopted in the endoscopic resection of soft tissue tumors in the parapharyngeal and lateral skull base, and the clinical application value of this method was evaluated. RESULTS: The blood loss during the operation was controlled within 150 mL, and the average blood loss was approximately 125 mL. The incidence of postoperative complications was low, and patients could recover well through functional training. The oral approach did not leave any wounds nor scars on the patient's facial skin after the operation and had no effect on the patient's appearance. CONCLUSIONS: The combination of 3D visualization technology, intraoperative real-time navigation, and endoscopy provides a beautiful, safe, and minimally invasive surgical method for patients with parapharyngeal or lateral skull base tumors.

Effects of Ilex latifolia and Camellia sinensis on cholesterol and circulating immune complexes in rats fed with a high-cholesterol diet.

Hypercholesterolaemia is one of the risk factors for atherosclerosis and subsequent cardiovascular disease. Here, we investigated the effects of dietary supplementation with Ilex latifolia or green tea (Camellia sinensis) on the levels of plasma total cholesterol, high-density lipoprotein cholesterol and circulating immune complexes in Sprague Dawley rats fed with a high-cholesterol diet. We demonstrated that daily administration by gavage of I. latifolia or C. sinensis at doses of 1.0 or 2.0 g/kg body weight for 30 days resulted in a significant decrease in plasma total cholesterol levels and circulating immune complexes and an increase in high-density lipoprotein cholesterol in rats fed with a high-cholesterol diet compared with levels in the high-cholesterol diet control group. C. sinensis was more effective than I. latifolia. I. latifolia and C. sinensis could be used as food supplements to protect against the development of hypercholesterolaemia.

Progress on the Role of Estrogen and Progesterone Signaling in Mouse Embryo Implantation and Decidualization.

Embryo implantation and decidualization are key steps in establishing a successful pregnancy. Defects in embryo implantation and decidualization can cause a series of adverse chain reactions which can contribute to harmful pregnancy outcomes, such as embryo growth retardation, preeclampsia, miscarriage, premature birth, and so on. Approximately 75% of failed pregnancies are considered to be due to embryo implantation failure or defects. Decidualization, characterized by proliferation and differentiation of uterine stromal cells, is one of the essential conditions for blastocyst implantation, placental formation, and maintenance of pregnancy and is indispensable for the establishment of pregnancy in many species. Embryo implantation and decidualization are closely regulated by estrogen and progesterone secreted by the ovaries. Many cellular events and molecular signaling network pathways are involved in this process. This article reviews the recent advances in the molecular mechanisms of estrogen- and progesterone-regulating uterine receptivity establishment, blastocyst implantation, and decidualization, in order to better understand the underlying molecular mechanisms of hormonal regulation of embryo implantation and to develop new strategies for preventing or treating embryo implantation defects and improving the pregnancy rate of women.

Multicenter Randomized Double-Blind Phase III Trial of Donafenib in Progressive Radioactive Iodine-Refractory Differentiated Thyroid Cancer.

PURPOSE: The phase II/III study of donafenib was initiated when there was no available treatment indicated for Chinese patients with progressive radioactive iodine-refractory differentiated thyroid cancer (RAIR-DTC). Donafenib, an oral tyrosine kinase inhibitor (TKI), showed good efficacy and tolerability in the phase II study. We aimed to further evaluate the antitumor activity and safety of donafenib in Chinese patients with RAIR-DTC. PATIENTS AND METHODS: This multicenter, double-blind, placebo-controlled, phase III study enrolled 191 patients with progressive RAIR-DTC and randomized in a ratio of 2:1 to donafenib (300 mg twice daily, n = 128) or matched placebo (n = 63). An open-label donafenib treatment period was allowed upon disease progression. The primary endpoint was progression-free survival (PFS) assessed by the independent review committee. The second endpoints include objective response rate (ORR), disease control rate (DCR), safety, etc. RESULTS: Donafenib demonstrated prolonged median PFS over placebo [12.9 vs. 6.4 months; hazard ratio (HR), 0.39; 95% confidence interval (CI), 0.25-0.61; P < 0.0001] in Chinese patients with RAIR-DTC. Improved ORR (23.3% vs. 1.7%; P = 0.0002) and DCR (93.3% vs. 79.3%; P = 0.0044) were observed in the donafenib group over placebo. For donafenib, the most common grade ≥ 3 treatment-related adverse events (AE) included hypertension (13.3%) and hand-foot syndrome (12.5%), 42.2% underwent dose reduction or interruption, and 6.3% experienced discontinuation. CONCLUSIONS: Donafenib was well-tolerated and demonstrated clinical benefit in terms of improved PFS, ORR, and DCR in patients with RAIR-DTC. The results suggest that donafenib could be a new treatment option for patients with RAIR-DTC.

miR-491-5p inhibits Emilin 1 to promote fibroblasts proliferation and fibrosis in gluteal muscle contracture via TGF-Beta1/Smad2 pathway.

Gluteal muscle contracture (GMC) is a chronic fibrotic disease of gluteal muscles due to multiple etiologies. Emilin 1 plays a determinant role in fibers formation, but its role in the progression of GMC remains unclear. The present study was aimed to search for the predictive role and regulatory mechanism of Emilin 1 on GMC. Here, Protein and mRNA expression of Emilin 1 were decreased in GMC tissues compared to normal muscle tissues. Using the anslysis of target prediction, Emilin 1 was observed to be a potential downstream sponge of miR-491-5p. In comparison to Emilin 1, miR-491-5p showed a aberrant elevation in GMC tissues, which was further proven to have a negative correlation with Emilin 1. The direct binding of miR-491-5p to Emilin 1 mRNA was confirmed by luciferase reporter gene assay, and miR-491-5p mimics inhibited, while miR-491-5p inhibitor promoted the protein expression and secretion of Emilin 1 in contraction bands (CB) fibroblasts. Additionally, miR-491-5p mimics promoted the expression of cyclin-dependent kinase 2 and cyclin D1 and the proliferation of CB fibroblasts, which could be reversed by Emilin 1 overexpression. Mechanistically, miR-491-5p mimics possibly activated transforming growth factor beta1 (TGF-beta1)/Smad3 signal cascade via binding to 3'-untranslated region of Emilin 1 mRNA, thereby promoting the progression of fibrosis of CB fibroblasts. Collectively, miR-491-5p inhibited Emilin 1 expression, and subsequently promoted CB fibroblasts proliferation and fibrosis via activating TGF-beta1/Smad3 signal axis. MiR-491-5p might be a potentially effective biomarker for predicting GMC, providing a novel therapeutic strategy for GMC.

Liver-specific overexpression of Gab2 accelerates hepatocellular carcinoma progression by activating immunosuppression of myeloid-derived suppressor cells.

GRB2-associated-binding protein 2 (Gab2) deletion has a preventive effect of on chronic liver inflammation and hepatocellular carcinoma. This study was aimed to elaborate Gab2-initiated immunoregulation during hepatocarcinogenesis. Compared to wild-type group, liver-specific overexpression of Gab2 mice (L-Gab2) displayed early hepatocarcinogenesis after 5-month diethylnitrosamine (DEN) induction, and accelerated tumor growth after 9-month DEN challenge. More myeloid-derived suppressor cells (MDSCs) were observed in DEN-challenged L-Gab2 mice than that in DEN-treated wild-type mice. Additionally, MDSCs activation-induced tumor angiogenesis capability and immunosuppression function were exceedingly activated in DEN-exposed L-Gab2 mice, which reflected in the increased platelet endothelial cell adhesion molecule (PECAM) and vascular endothelial growth factor (VEGF), and the decreased cytotoxic T lymphocytes. Mechanistically, DEN-challenged L-Gab2 mice produced more IL-6, and IL-6 depletion significantly deprived Gab2-overexpression-mediated tumor-promotion phenomena, accompanied by the impairment of MDSCs-initiated immunosuppression function. MDSCs isolated from IL-6-depleted L-Gab2 mice or inactivating MDSCs partly restored the immune function of cytotoxic T cells. Of note, MDSCs gene signatures had a significant association with the increased Gab2 or IL6 in hepatoma specimens. Collectively, L-Gab2 mice accelerated hepatoma progression possibly through activating IL-6-initiated the activation of MDSCs. This study provides a novel insights for exploring the role of Gab2 in autoimmune tolerance during hepatocarcinogenesis.

L22 ribosomal protein is involved in dynamin-related protein 1-mediated gastric carcinoma progression.

Mitochondrial fission depends on dynamin-related protein 1 (Drp1) guanosine triphosphatase activity. Although there is some association between Drp1 and gastric cancer, the detailed mechanism remains largely unknown. In this study, the elevation of Drp1 was observed in human gastric carcinoma specimens including gastric mixed adenocarcinoma tissues, gastric intestinal-type adenocarcinoma tissues, and human gastric cancer cells compared to normal control, but not in diffuse gastric adenocarcinoma tissues. Gastric cancer patients with high Drp1 harbored advanced pathological stages and poor progression-free survival probability compared to those with low Drp1. Mdivi-1-mediated inactivation of Drp1 robustly inhibited cell viability and tumor growth but conversely induced cell apoptotic events in vitro and in vivo. Based on the Encyclopedia of RNA Interactomes Starbase, L22 ribosomal protein (RPL22) was recognized as the potential downstream oncogene of Drp1. Clinically, the significant correlation of Drp1 and RPL22 was also verified. Mechanistically, Drp1 inactivation did not affect the accumulation of RPL22 in gastric carcinoma. However, the intracellular distribution of RPL22 had an endonuclear location in Drp1-inactivated tumors. Of note, Drp1 inactivation notably reduced the expression of cytoplasmic RPL22 and increased its nuclear level in gastric cancer cells. Collectively, Drp1 had high levels in human gastric carcinoma specimens and could serve as a potential diagnostic and prognostic biomarker in gastric carcinoma. The Drp1 inactivation-mediated anti-proliferative and pro-apoptosis effects on gastric cancer were possibly associated with nuclear import of RPL22. This knowledge may provide new therapeutic tools for treating gastric carcinoma via targeting mitochondria-related ribosome pathway.

Pharmacokinetics and bioavailability of cefquinome in healthy ducks.

OBJECTIVE: To determine pharmacokinetics and bioavailability of cefquinome administered IV, IM, or PO to healthy ducks. ANIMALS: Thirty-six 2-month-old Muscovy ducks. PROCEDURES: Ducks were randomly assigned to 3 groups of 12 birds each for a single IV, IM, or PO administration at a dose of 5 mg/kg. Blood samples were collected before and at various intervals after each administration. Cefquinome concentration was determined by use of high-performance liquid chromatography at 268 nm with a UV detector, and pharmacokinetics were analyzed. RESULTS: The disposition of cefquinome following IV or IM administration was best described by a 2-compartment model. After IV administration, mean ± SD elimination halflife was 1.57 ± 0.06 hours, clearance value was 0.22 ± 0.02 L/kg·h, and apparent volume of distribution at steady state was 0.41 ± 0.04 L/kg. After IM administration, elimination half-life was 1.79 ± 0.13 hours, peak concentration time was 0.38 ± 0.06 hours, peak drug concentration was 9.38 ± 1.61 µg/mL, and absolute mean ± SD bioavailability was 93.28 ± 13.89%. No cefquinome was detected in plasma after PO administration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that cefquinome was absorbed quickly and had excellent bioavailability after IM administration, but absorption after PO administration was poor.

Estrogen receptor α is involved in the regulation of ITGA8 methylation in estrogen receptor-positive breast cancer.

BACKGROUND: Integrin subunit α 8 (ITGA8) methylation has been associated with the development of several cancers, but its contribution to breast cancer remains unclear. The present study aimed to investigate the methylation status of ITGA8, and the underlying regulatory mechanisms of ITGA8 methylation in breast cancer. METHODS: ITGA8 expression was investigated using the Gene Expression Profiling Interactive Analysis 2 (GEPIA2) database and the Breast Cancer Gene-Expression Miner v.4.4 (bc-GenExMiner v4.4). The association between ITGA8 expression levels and the survival rate of breast cancer patients was evaluated using The Cancer Genome Atlas (TCGA) database and Gene Expression-based Outcome for Breast Cancer Online (GOBO): Gene Set Analysis. Methylation-specific PCR (MSP) was used to detect the methylation of ITGA8. Protein level of ITGA8 was determined by Western blot analysis. RESULTS: ITGA8 was expressed at low levels in human breast cancer cells compared to non-tumorigenic breast cells and breast tissue, and was upregulated in estrogen receptor (ER)-positive tissue compared with ER-negative tissue (P<0.01). ITGA8 gene expression was negatively associated with breast tumor stage and survival rate in all breast cancer patients. However, ER-positive patients with low ITGA8 expression showed poorer distant metastasis-free survival (DMFS) and recurrence-free survival (RFS) rates than patients with high ITGA8 expression. This was not observed in the ER-negative population. Mechanistically speaking, hypermethylation of ITGA8 was discovered in ER-positive breast cancer cells. Administration of the methylation inhibitor, 5-aza-2'-deoxycytidine (5-aza-dC), significantly elevated protein expression of ITGA8 in ER-positive breast cancer cells compared to ER-negative cells. The positive association between ITGA8 status and methylation was also observed in clinical tissue specimens. When treated with 17-beta-estradiol, an antagonist of ERα, 5-aza-dC-induced upregulation of ITGA8 in ER-positive breast cancer cells was no longer observed. CONCLUSIONS: Low ITGA8 expression in ER-positive breast cancer might be caused by the hypermethylation of ITGA8, a process dependent on ERα. Our findings provide an important foundation for investigations into ITGA8-targeted treatment strategies for ER-positive breast cancer.