Hypoxia and aberrant activation of epidermal growth factor receptor (EGFR) are considered important features of various malignancies. However, whether hypoxia can directly trigger EGFR activation and its clinical implications remain unclear. In this study, we demonstrated that in oral cancer, a typical hypoxic tumor, hypoxia can induce chronic but constitutive phosphorylation of wild-type EGFR in the absence of ligands. Oral cancer cell lines exhibit different EGFR phosphorylation responses to hypoxia. In hypoxic HN4 and HN6 cells, ubiquitination-mediated endocytosis, lysosomal sorting, and degradation lead to low levels of EGFR phosphorylation. However, in CAL-27 and HN30 cells, a novel HIF-1α-induced long noncoding RNA (lncRNA), EUDAL, can compete with the E3 ligase/adaptor complex c-Cbl/Grb2 for binding to EGFR, stabilizing phosphorylated EGFR (pEGFR) and resulting in sustained activation of EGFR and its downstream STAT3/BNIP3 signaling. STAT3/BNIP3-mediated autophagy leads to antitumor drug resistance. A high EUDAL/EGFR/STAT3/autophagy pathway activation predicts poor response to chemotherapy in oral cancer patients. Collectively, hypoxia can induce noncanonical ligand-independent EGFR phosphorylation. High EUDAL expression facilitates sustained EGFR phosphorylation in hypoxic tumor cells and leads to autophagy-related drug resistance.
Humans ; ErbB Receptors/metabolism* ; Mouth Neoplasms/pathology* ; RNA, Long Noncoding/genetics* ; Drug Resistance, Neoplasm/genetics* ; Cell Line, Tumor ; Phosphorylation ; Signal Transduction ; STAT3 Transcription Factor/metabolism* ; Cell Hypoxia ; Autophagy ; Proto-Oncogene Proteins c-cbl/metabolism*

To analyze the current status and epidemiological trends of the disease burden of gastric cancer among adolescents and young adults in China, so as to provide a basis for formulating public health policies and reducing the disease burden.

Lymphoedema of lower extremities,chronic and progressive,will severely deteriorate the quality of life of patients as it progresses.Thus,early diagnosis and treatment to delay the progress of the disease is conducive to improving the prognosis of patients.At present,common techniques for the diagnosis of lower limb lymphedema,whose advantages and disadvantages vary,cannot be applied to individual case comprehensively.CEUS has the advantages of non-invasion,convenience,real-time,and good repeatability for this disease.CEUS can enhance the image of lymph in lymphatics,and has a high sensitivity to superficial lymphatics,gradually applied in lymphedema of lower limbs.This article reviews the application of CEUS in lower limb lymphedema.

Chronic aortic stenosis and regurgitation can cause left ventricular remodeling.Whether these changes are reversible and their reversibility after valve replacement are the main determinants affecting the timing and prognosis of surgery.Imaging techniques are commonly used to evaluate myocardial structure and function,in which echocardiography and enhanced CT are helpful to evaluate artificial flap function and monitor left ventricular deformation,while cardiac MR and PET/CT are helpful to identify the progression and regression of postoperative cardiac fibrosis.The combined application of these new techniques can improve clinical outcomes by early diagnosis and non-invasive detection of postoperative left ventricular reverse remodeling.This paper reviews the evaluation and application of multi-modal imaging techniques for left ventricular reverse remodeling after aortic valve replacement.

Objective To explore the role of TFF3 in lung adenocarcinoma using bioinformatics methods and in vitro cell experi-ments.Methods We analyzed the characteristic differences between patient groups with high and low TFF3 expression using the TCGA database and R package.The analysis covered various perspectives,including clinical features,immune microenvironment,tumor muta-tion burden,and immunotherapy.The effect of TFF3 on the number of NCI-H1975 cells was determined using RTCA,while the expres-sion of the PI3K/Akt signaling pathway Akt and its activated state p-Akt was detected using Western blot.Results The expression of TFF3 increased in lung adenocarcinoma tissue,and this increase was related to the pathological type of lung adenocarcinoma.However,it was not associated with the prognosis of patients.Low expression of TFF3 was associated with low tumor purity.Additionally,TFF3 showed a negative correlation with tumor mutation burden and the expression of several immune checkpoint genes.In our study,we treated NCI-H1975 cells with TFF3 and observed an increase in the total number of cells.Furthermore,the results from Western blot analysis indicated that TFF3 could activate the expression of the p-Akt gene.Conclusion TFF3 could be upregulated in specific subtypes of lung adenocarcinoma and linked with tumor immunity of lung adenocarcinoma.We observed that TFF3 can enhance the growth of lung ad-enocarcinoma cells,possibly by activating the PI3K/Akt signaling pathway.Our findings suggest that TFF3 could serve as a promising therapeutic target for individuals diagnosed with lung adenocarcinoma.

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Oral mucosal melanoma (OMM), a subtype of melanoma which is commonly found in the eastern Asian populations, progresses with unclear pathogenesis, high malignancy and poor prognosis. Constructing different types of preclinical models for OMM, which simulate clinical characteristics such as tumor invasion and metastasis, assists screening and efficacy-evaluation of drugs. This would promote personalized treatments for patients with OMM. However, lack of preclinical models makes one of the critical obstacles that hinder the recognition of mucosal melanoma and block the treatment breakthrough in mucosal melanoma. In recent years, certain progress has been made in the construction and application of OMM preclinical models. Various OMM preclinical models have been successfully constructed and carried out for further research, assisting in excavating personalized treatment strategies. In this review, we will summarize the latest progress in the researches on OMM preclinical models.

Objective:To investigate the physicochemical and biological properties of different magnesium modified calcium phosphate bone cements.Methods:The different magnesium modified calcium phosphate bone cements were divided into magnesium citrate, magnesium lactate, magnesium malate, magnesium phosphate and magnesium glycinate groups, each of which was added with different magnesium agents in the proportion of 0%, 1%, 3% and 5% of the total weight of calcium phosphate bone cements. The initial and final setting time, injectability, anti-collapse performance and compressive strength of different magnesium modified calcium phosphate bone cements were tested. Furthermore, the screened bone cement extracts were used to culture with third generation osteoblasts. Bioactivity assays were performed using the Cell Proliferation and Toxicity Assay Kit (CCK-8). Alkaline phosphatase (ALP) staining and Alizarin Red S (ARS) staining were performed on osteoblasts to observe the osteogenic activity of magnesium malate modified calcium phosphate bone cements.Results:The addition of different proportions of different magnesium agents led to the shortening of the initial and final setting time of modified calcium phosphate bone cements. Moreover, the final setting time of 5% magnesium malate modified calcium phosphate bone cements was the shortest (<40 minutes), which was significantly shorter compared with other magnesium agents in the same proportion (all P<0.05). With the addition of different magnesium agents in different proportions, the injectability of bone cements was gradually increased, and the injectability of 5% magnesium malate calcium phosphate bone cements reached the highest for (87.3±1.9)%, which was significantly increased compared with other magnesium agents in the same proportion (all P<0.05). The anti-collapse performance of bone cements was decreased with the addition of different magnesium agents in different proportions. Magnesium citrate, magnesium phosphate and magnesium glycinate modified calcium phosphate bone cements could not resist the flushing of deionized water. In particular, magnesium malate modified calcium phosphate bone cements had the best anti-collapse performance, with the maximum weight loss rate for only (9.8±2.3)% after 30 minutes of deionized water flushing, which was better than the rest of the groups (all P<0.05). The compressive strength of magnesium lactate and magnesium phosphate modified calcium phosphate bone cements showed a decrease compared with original calcium phosphate bone cements, while the compressive strength of magnesium citrate and magnesium malate modified calcium phosphate bone cements was significantly increased compared with original calcium phosphate bone cements, of which 3% magnesium malate modified calcium phosphate bone cements had the greatest compressive strength of (6.2±0.2)MPa, significantly higher than the rest of the groups (all P<0.05). The sieve test yielded magnesium malate modified calcium phosphate bone cement, which had a weight loss of (27.0±0.9)% at 35 days in vitro. The release of magnesium ions was increased with increasing magnesium malate dose in the in vitro environment of magnesium malate modified calcium phosphate bone cements in different ratios. A stable magnesium ion release was achieved within 35 days.Also, the pro-proliferative and osteogenic effects of modified calcium phosphate bone cements on osteoblasts were more obvious with increase of magnesium malate dose. For 5% magnesium malate modified calcium phosphate bone cements, the cell number, ALP staining area ratio and calcium nodule area ratio were significantly increased compared with the groups in the proportion of 0% and 1% magnesium malate (all P<0.05). Conclusions:Among magnesium citrate, magnesium lactate, magnesium malate, magnesium phosphate and magnesium glycinate modified calcium phosphate bone cements, magnesium malate modified calcium phosphate bone cements have relatively suitable setting time, excellent anti-collapse performance and mechanical strength. Meanwhile, 5% magnesium malate modified calcium phosphate bone cements have better biological activity among different ratios of magnesium malate modified calcium phosphate bone cements, suggesting a potential value for clinical application.

Objective:To investigate the effect of Asarinin on the survival time of transplanted heart after allogeneic heterotopic heart transplantation and to further verify the anti-immune rejection effect of Asarinin in spleen and peripheral blood.Methods:Using 64 Wistar rats as donors, 64 SD rats as recipients to establish the allogeneic heterotopic heart transplantation model in rats.After successful transplantation, 64 rats were use simple randomization divided into control group, cyclosporine A(CsA) group, Asarinin group and half CsA + half Asarinin group with 16 rats in each group.CsA group was given 5 mg/kg by gavage; Asarinin group was given 25 mg/kg; half dose group was given CsA 2.5 mg/kg+ Asarinin 12.5 mg/kg and the control group was given the same volume of normal saline by gavage.After administration for 1 week, half of them were used to observe the survival time.The other half of the rats were fully anesthetized with chloral hydrate, spleen and peripheral blood were taken.Half of the spleen was taken to observe the slices under the microscope.The other half of spleen was used RT-PCR to detect the relative expression of IFN-γ and IL-4.The expression of co-stimulatory molecules CD80, CD86 and CD40 in peripheral blood were detected by flow cytometry.Results:Survival time of transplanted heart was control group (8.4±0.9), CsA group (30.5±8.3), Asarinin group (16.5±4.3) and half-dose group (26.1±5.2) days.Compared with control group, survival time of heart transplantation became prolonged in all groups and the difference was statistically significant ( P<0.05). HE staining of splenic tissue showed that, as compared with control group, the injury of each group was alleviated.The relative expression of IFN-γ in spleen was control group (1.055±0.083), CsA group (0.396±0.038), Asarinin group (0.833±0.094) and half-dose group (0.862±0.104). The last three groups were lower than control group and the difference was statistically significant ( P<0.05). The relative expression of IL-4 in spleen was control group (1.429±0.234), CsA group (3.808±0.729), Asarinin group (2.209±0.306) and half-dose group (2.323±0.321). The last three groups all spiked as compared with control group and the difference was statistically significant ( P<0.05). The expressions of CD80, CD86 and CD40 in peripheral blood were control group (98.21±0.54), (85.78±0.89) and (96.36±0.66), CsA group (89.26±0.36), (56.86±2.32) and (88.11±1.61), Asarinin group (94.19±0.47), (79.01±1.12) and (87.86±1.67) and half-dose group (94.87±0.74), (80.81±0.98) and (89.71±0.97) respectively.The last three groups were lower than control group and the difference was statistically significant ( P<0.05). Conclusions:Asarinin can prolong the survival time of transplanted heart after allogeneic heterotopic heart transplantation in rats, inhibit the immune injury of spleen after allogeneic heterotopic heart transplantation in rats, decrease IFN-γ in spleen, increase IL-4 in spleen and inhibit the expression of peripheral blood costimulatory molecules CD80, CD86 and CD40.

The community structure and diversity of the gut microbiota are associated with human diseases. However, the analysis of different community structure might be influenced by experimental approaches such as the quality of DNA extraction. Therefore, evaluating the efficiency of different DNA extraction methods for specific intestinal species is a guideline for obtaining a comprehensive human gut microbial profile, which may assist the in-depth investigation into the structure of the gut microbial community. The aim of this study was to perform a comparative analysis of five different DNA extraction methods. With the aid of qPCR, the efficiency of five DNA extraction kits was evaluated in terms of the purity of the extracted DNA, the DNA concentration, and the abundance of genomic DNA extracted from specific intestinal species. The results showed that the kit Q gave the best extraction results, especially for Gram-positive bacteria such as Lactobacillus and Bifidobacterium. The average DNA concentration of the N kit was lower than that of the Q kit, but there was no significant difference between the two in terms of the purity. Compared to the other three commercial kits (M, PSP, TG), the efficiency of the N kit in extracting the genomic DNA of the specified microorganisms were the least different from those of the Q kit. In contrast, the DNA extracted by the M kit was of higher quality but of lower concentration, and was not very efficient for Gram-positive bacteria. The DNA extracted by the TG and PSP kits was inferior to the other validated kits in terms of the concentration, quality and bacterial abundance. These results provide a basis for the selection of genomic DNA extraction methods in microecological research experiments.
DNA/genetics* ; DNA, Bacterial/genetics* ; Feces/microbiology* ; Humans ; Microbiota/genetics* ; RNA, Ribosomal, 16S/genetics*