Effects of different addition levels of CHM-JM113 on growth performance, antioxidant capacity, organ index, and intestinal health of AA broilers.

The purpose of the present study was to investigate the effects of different levels of a Chinese herbal medicine formulation combined with JM113 (CHM-JM113) on growth performance, antioxidant capacity, organ index, and intestinal health of AA broilers. The AA broiler chicks were randomly allocated to 5 treatments as follows: a basic diet for the control group, the basic diet supplemented with 0.25% CHM-JM113, 0.5% CHM-JM113, 1% CHM-JM113 and 2% CHM-JM113 for the treatment group, respectively. The results showed that the addition of CHM-JM113 to the diet significantly reduced the mortality (p < 0.01) and improved the European Broiler Index (EBI) (p < 0.05), whereas it had no significance on growth performance of AA broilers (p > 0.05). Comparing the control group, 0.5 and 1% CHM-JM113 group significantly improved the organ index of liver, spleen and bursa (p < 0.05). In terms of intestinal morphology and structure, the addition of different levels of CHM-JM113 increased VH and VH/CD ratio, decreased CD in the small intestine compared to the control group, with 1 and 2% of the additive dose being more effective (p < 0.05). Chinese herbal medicine and probiotics as natural antioxidants also significantly increased the content of SOD in serum of 21-day-old broilers (p < 0.01), and significantly decreased the content of MDA in serum (p < 0.01). At 42 days of age, the addition of 1 and 2% CHM-JM113 significantly increased the content of SOD (p < 0.01) and significantly decreased the content of MDA in the organism (p < 0.01), accompanied by a significant increase in T-AOC and CAT content. In the study of the effect of CHM-JM113 on intestinal immunity, compared with the control group, we found that 1% or 2% CHM-JM113 had a better effect on the expression of occludin and claudin-1 in the intestinal segments of broilers (p < 0.05). For the expression of GATA-3, 0.5% CHM-JM113 may have a better effect (p < 0.05). CHM-JM113 may be used as an antibiotic alternative in broiler production.

Cell Membrane-Coated Biomimetic Nanoparticles in Cancer Treatment.

Nanoparticle-based drug delivery systems hold promise for cancer treatment by enhancing the solubility and stability of anti-tumor drugs. Nonetheless, the challenges of inadequate targeting and limited biocompatibility persist. In recent years, cell membrane nano-biomimetic drug delivery systems have emerged as a focal point of research and development, due to their exceptional traits, including precise targeting, low toxicity, and good biocompatibility. This review outlines the categorization and advantages of cell membrane bionic nano-delivery systems, provides an introduction to preparation methods, and assesses their applications in cancer treatment, including chemotherapy, gene therapy, immunotherapy, photodynamic therapy, photothermal therapy, and combination therapy. Notably, the review delves into the challenges in the application of various cell membrane bionic nano-delivery systems and identifies opportunities for future advancement. Embracing cell membrane-coated biomimetic nanoparticles presents a novel and unparalleled avenue for personalized tumor therapy.

Photoredox/Copper Dual-Catalyzed Benzylic C-H Esterification via Radical-Polar Crossover.

Herein, we report a distinctive photoredox/copper dual-catalyzed esterification of benzylic C-H bonds through the combination of photoredox-mediated hydrogen atom transfer and Cu(II)-mediated radical-polar crossover. This methodology demonstrates a high functional group tolerance (>40 examples) and moderate to good yields with structurally diverse benzylic C-H substrates. Notably, stoichiometric amounts of carboxylic acids are used as coupling partners, which allows the synthesis of structurally diverse benzylic esters and the late-stage functionalization of pharmaceuticals.

Early Prediction of Refractory Epilepsy in Children Under Artificial Intelligence Neural Network.

In order to realize the early prediction of refractory epilepsy in children, data preprocessing technology was used to improve the data quality, and the detection model of refractory epilepsy in children based on convolutional neural network (CNN) was established. Then, the data in the epilepsy electroencephalography (EEG) signal public data set was used for model training and the diagnosis of refractory epilepsy in children. Moreover, back propagation neural network (BPNN), support vector machine (SVM), XGBoost, gradient boosting decision tree (GBDT), AdaBoost algorithm were introduced for comparison. The results showed that the early prediction accuracy of BP, SVM, XGBoost, GBDT, AdaBoost, and the algorithm in this study for refractory epilepsy in children were 0.745, 0.778, 0.885, 0.846, 0.874, and 0.941, respectively. The sensitivities were 0.81, 0.826, 0.822, 0.84, 0.859, and 0.918, respectively. The specificities were 0.683, 0.696, 0.743, 0.792, 0.84, and 0.905, respectively. The accuracy was 0.707, 0.732, 0.765, 0.802, 0.839, and 0.881, respectively. The recall rates were 0.69, 0.716, 0.753, 0.784, 0.813, and 0.877, respectively. F1 scores were 0.698, 0.724, 0.759, 0.793, 0.826, and 0.879, respectively. Through the comparisons of the above six indicators, the algorithm proposed in this study was significantly higher than other algorithms, suggesting that the proposed algorithm was more accurate in early prediction of refractory epilepsy in children. Analysis of the EEG characteristics and magnetic resonance imaging (MRI) images of refractory epilepsy in children suggested that the MRI images of patients' brains under this algorithm had obvious characteristics. The reason for the prediction error of the algorithm was that the duration of epilepsy was too short or the EEG of the patient didn't change notably during the epileptic seizure. In summary, the prediction method of refractory epilepsy in children based on CNN was accurate, which had broad adoption prospects in assisting clinicians in the examination and diagnosis of refractory epilepsy in children.

Exosomes containing miR-122-5p secreted by LPS-induced neutrophils regulate the apoptosis and permeability of brain microvascular endothelial cells by targeting OCLN.

OBJECTIVE: To explore the effect of exosomes containing miR-122-5p secreted by lipopolysaccharide (LPS)-induced neutrophils on the apoptosis and permeability of brain microvascular endothelial cells (BMECs). METHODS: Neutrophils in blood were isolated, purified and identified. LPS-induced neutrophils were co-cultured with BMECs. Untreated or LPS-induced neutrophil exosomes were isolated and identified with a transmission electron microscope. miR-122-5p expressions in the exosomes were detected by real-time quantitative polymerase chain reaction, and then the exosomes were co-cultured with BMECs. Bioinformatics analysis was performed to predict the downstream target gene of miR-122-5p, and OCLN was selected as the subject. Dual luciferase reporter assay was carried out to verify the interactive relationship between OCLN and miR-122-5p. LPS and miR-122-5p were used to treat neutrophils, and then exosomes were collected. Exosome or OCLN was embedded in BMECs. The proliferation, colony forming ability and apoptosis of BMECs were detected by cholecystokinin octopeptide, clone formation assay and flow cytometry, respectively. Corresponding kits were used to detect the activities of reactive oxygen species, superoxide dismutase, malondialdehyde and catalase. Vascular endothelial growth factor and tight junction proteins (ZO-1 and Claudin-5) expressions were measured by Western blot for cell permeability evaluation. RESULTS: miR-122-5p had an increased expression in LPS-induced neutrophil exosomes and could promote oxidative stress, apoptosis and permeability increase of BMECs and the inhibition of BMECs proliferation and colony formation (P<0.05). miR-122-5p targeted the binding with OCLN and down-regulated OCLN expression. OCLN overexpression partly decreased the malignant effect of miR-122-5p on BMECs (P<0.05). CONCLUSION: LPS can induce neutrophils to secrete exosomes containing miR-122-5p. The down-regulation of OLCN expression can aggravate BMECs injury.

MicroRNA miR-126 attenuates brain injury in septic rats via NF-κB signaling pathway.

The purpose of this study was to investigate the impact and mechanism of microRNA miR-126 on brain injury induced by blood-brain barrier (BBB) damage in septic rats. We used cecal ligation and perforation (CLP) to create a rat model of sepsis. The experimental rats were randomly divided into Control group, CLP group, CLP + miR-NC group, CLP + miR-126 group and CLP + miR-126 + NF-κB pathway agonist (PMA) group. MiR-126 expressed in the brain tissue of CLP rats was down-regulated by qRT-PCR. Upregulation of miR-126 in CLP rats could improve brain injury and BBB marker protein level, reduce brain water content, Evans blue extravasation, inflammation, and excessive oxidative stress. This could also result in an inhibition of NF-κB signaling pathway activity. In conclusion, miR-126 overexpression can prevent brain injury caused by BBB damage via the inhibition of NF-κB signaling pathway activity.

Genomic Feature of a Rare Case of Mix Small-Cell and Large-Cell Neuroendocrine Lung Carcinoma: A Case Report.

BACKGROUND: Cases of both of small- (SCLC) and large-cell neuroendocrine lung carcinoma (LCNEC) were rarely reported. Although typical cases are morphologically distinct, the distinction between LCNEC and SCLC is still controversial, with some LCNECs showing close morphologies with SCLC. Here, we reported on a patient who had tumor with a mix of SCLC and LCNEC and uncovered these components' histological and genomic features. CASE PRESENTATION: A 59-year-old man was diagnosed with lung cancer and had resection surgery in our hospital. The H&E and immunohistochemistry staining revealed that the tumor had 30%-35% LCNEC and 65%-70% SCLC cells. The whole-exome sequencing (WES) identified no potentially actionable alteration in the tumor sample but found five alterations all with allele frequency over 90%, including TP53 p.R273H, MYH8 p.Q1814K, SLC17A6 p.W505L, PTPN5 p.M40I, and RB1 p.L267X. The genomic results supported that these two different components shared a similar dominant clonal origin. Furthermore, fluorescence in situ hybridization analysis revealed that the LCNECs have a higher copy number of MET than the SCLC component while without notable difference in the copy number of HER2 and TP53. Chemotherapy with pemetrexed and carboplatin was administrated for two cycles after the surgery. Although the chest CT showed remission in the lung, he was diagnosed with bone metastasis in 1 year later. Then, he received chemotherapy with etoposide and carboplatin but had severe side effect, leading to the discontinuation of the regime. Unfortunately, he returned to the local hospital with supportive care and died shortly after. CONCLUSION: Based on these observations, we proposed that LCNEC and SCLC components in this patient may have a common clonal origin with dual mutations in TP53 and RB1, while the chromosome instability may cause multiple independent conversion that leads to LCNEC or SCLC morphologies.

WITHDRAWN: Analysis of Risk Factors for Prognosis and Infection of Child with Refractory Epilepsy Via Artificial Intelligence Neural Network Image Information.

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EMX2 is epigenetically silenced and suppresses epithelial­mesenchymal transition in human esophageal adenocarcinoma.

Esophageal adenocarcinoma (EAC) is an aggressive and challenging disease to treat, with an overall five­year survival rate of <20%. Early malignant cell dissemination contributes to this poor prognosis. Epithelial­mesenchymal transition (EMT) induces the invasion and metastasis of carcinoma cells. Empty spiracles homeobox 2 (EMX2) is a homeodomain­containing transcription factor, which is associated with numerous cancer types, and has been demonstrated to regulate EMT. In the present study, 48 pairs of EAC and adjacent normal tissues were analyzed. The results revealed that EMX2 was downregulated in EAC tissues, and its expression was negatively correlated with the DNA hypermethylation of its promoter. Additionally, the OE19 and OE33 EAC cell lines were treated with the DNA methyltransferase inhibitor 5­aza­2'­deoxycytidine, and the results indicated that EMX2 expression was increased. Overexpressing EMX2 in EAC cell lines enhanced the expression of apoptotic markers, inhibited cell migration and invasion, led to the upregulation of E­cadherin and the downregulation of mesenchymal markers, and suppressed AKT, mTOR and S6K phosphorylation. Furthermore, EMX2 overexpression sensitized EAC cells to cisplatin. These results demonstrated that EMX2 inhibited the AKT/mTOR/S6K signaling pathway and decreased EMT. However, the downregulation of EMX2 was revealed to be associated with EMT in EAC, indicating that EMX2 may be a potential target for the management of EAC.

A model for the impact of FFPE section thickness on gene copy number measurement by FISH.

Fluorescent in situ hybridization (FISH) assays to detect gene amplification such as HER2 or MET in tumors are used for prognosis evaluation and selection of targeted therapies. Although FISH guidelines recommended 4~6 µm FFPE sections, many laboratories use 2~3 µm sections, which is a common practice for H&E staining and immunohistochemistry. A former study concluded that section thickness did not affect FISH results. We found, however, that thinner FFPE sections may lead to false negative results for gene amplification. A mathematic model was constructed and cell-line based controls with known gene copy number were prepared, and the model had a reasonable fit with the experimental data. The model revealed that even when counting the apparently full-sized nuclear images, many of them have partial volumes, which leads to under-estimation of gene copy number. Therefore, improperly thinner sections are prone to give false negative results, and thicker sections give a better approximation to the true value. The discrepancy between this and the former study was discussed. In summary, the model applies generally to FISH/ISH detection of gene copy number, and section thickness is an important parameter to control for precision medicine research, assay development, clinical trials and daily practice in pathology laboratory.

Solanum nigrum polysaccharide inhibits tumor growth in H22-bearing mice through regulation of caspase-3 and bcl-2.

OBJECTIVE: The objective of this study was to investigate the effect of Solanum nigrum polysaccharides (SNPs) on tumor growth in H22 hepatocarcinoma cells bearing mice and explore the mechanism by focusing on the regulation of the expression of caspase-3 and bcl-2. MATERIALS AND METHODS: Totally, 50 mice bearing with H22 cells were randomly divided into five groups: Model group, cyclophosphamide group (CTX, 30 mg/kg), SNP groups with low, medium, and high doses of SNP (30, 60, and 120 mg/kg). Twenty-four hours after inoculation of H22 cells, CTX or SNP were given by gavage once a day for 10 days. The growth of tumor was observed. The tumor inhibition rate, indexes of the spleen and thymus were calculated. The immunohistochemical method was used for the determination of caspase-3 and bcl-2 expression in the tumor tissue. RESULTS: SNP (30, 60, and 120 mg/kg) reduced the average tumor weight compared with that in model group in a dose-dependent manner, and the tumor inhibition rates were 37.73%, 38.24%, and 42.60%, respectively. In addition, SNP dose dependently increased the index of the thymus compared with that of the CTX group. Immunohistochemistry results showed that the protein expression of caspase-3 in SNP groups was higher, but the expression of bcl-2 was lower than that in model group in a dose-dependent manner. CONCLUSION: SNP inhibited the growth of tumor in H22-bearing mice and protected the immune organ. The mechanism underlying the inhibition of tumor might be related to the upregulation of caspase-3 and downregulation of bcl-2.

DR5 mAb-conjugated, DTIC-loaded immuno-nanoparticles effectively and specifically kill malignant melanoma cells in vivo.

We combined chemo- and immunotherapies by constructing dual therapeutic function immuno-nanoparticles (NPs) consisting of death receptor 5 monoclonal antibody (DR5 mAb)-conjugated nanoparticles loaded with dacarbazine (DTIC) (DTIC-NPs-DR5 mAb). We determined the in vivo targeting specificity of DTIC-NPs-DR5 mAb by evaluating distribution in tumor-bearing nude mice using a real-time imaging system. Therapeutic efficacy was assessed in terms of its effect on tumor volume, survival time, histomorphology, microvessel density (MVD), and apoptotic index (AI). Systemic toxicity was evaluated by measuring white blood cells (WBC) counts, alanine aminotransferase (ALT) levels, and creatinine clearance (CR).In vivo and ex vivo imaging indicates that DR5 mAb modification enhanced the accumulation of NPs within the xenograft tumor. DTIC-NPs-DR5 mAb inhibited tumor growth more effectively than DTIC or DR5 mAb alone, indicating that combining DTIC and DR5 mAb through pharmaceutical engineering achieves a better therapeutic effect. Moreover, the toxicity of DTIC-NPs-DR5 mAb was much lower than that of DTIC, implying that DR5 mAb targeting reduces nonspecific uptake of DTIC into normal tissue and thus decreases toxic side effects. These results demonstrate that DTIC-NPs-DR5 mAb is a safe and effective nanoparticle formulation with the potential to improve the efficacy and specificity of melanoma treatment.

Peptide-Mediated Tumor Targeting by a Degradable Nano Gene Delivery Vector Based on Pluronic-Modified Polyethylenimine.

Polyethylenimine (PEI) is considered to be a promising non-viral gene delivery vector. To solve the toxicity versus efficacy and tumor-targeting challenges of PEI used as gene delivery vector, we constructed a novel non-viral vector DR5-TAT-modified Pluronic-PEI (Pluronic-PEI-DR5-TAT), which was based on the attachment of low-molecular-weight polyethylenimine (LMW-PEI) to the amphiphilic polymer Pluronic to prepare Pluronic-modified LMW-PEI (Pluronic-PEI). This was then conjugated to a multifunctional peptide containing a cell-penetrating peptide (TAT) and a synthetic peptide that would bind to DR5-a receptor that is overexpressed in cancer cells. The vector showed controlled degradation, favorable DNA condensation and protection performance. The Pluronic-PEI-DR5-TAT/DNA complexes at an N/P ratio of 15:1 were spherical nanoparticles of 122 ± 11.6 nm and a zeta potential of about 22 ± 2.8 mV. In vitro biological characterization results indicated that Pluronic-PEI-DR5-TAT/DNA complexes had a higher specificity for the DR5 receptor and were taken up more efficiently by tumor cells than normal cells, compared to complexes formed with PEI 25 kDa or Pluronic-PEI. Thus, the novel complexes showed much lower cytotoxicity to normal cells and higher gene transfection efficiency in tumor cells than that exhibited by PEI 25 kDa and Pluronic-PEI. In summary, our novel, degradable non-viral tumor-targeting vector is a promising candidate for use in gene therapy.