SPP1 promotes tumor progression in esophageal carcinoma by activating focal adhesion pathway.

Background: Recurrence and metastasis are the major obstacles affecting the therapeutic efficacy and clinical outcomes for patients with esophageal carcinoma (ESCA). Secreted phosphoprotein 1 (SPP1) is considered as a hub gene in ESCA and is negatively associated with disease-free survival (DFS) in ESCA. However, the exact roles and underlying mechanisms remain elusive. This study aims to examine the roles of SPP1 on ESCA, and elucidate the potential mechanisms. Methods: Bioinformatics were used to analyze the expression of SPP1 in ESCA tissues, and its relations with clinicopathological characteristics and clinical prognosis in patients with ESCA based on The Cancer Genome Atlas (TCGA) dataset. Loss-of-function was conducted to examine the roles of SPP1 on malignant behaviors of ESCA cells by cell counting kit-8 (CCK8), plate clone, wound healing, and transwell assays. Gene set enrichment analysis (GSEA) was conducted to screen the pathways associated with SPP1 in ESCA. Then, the enriched pathway and the underlying mechanism were elucidated by western blotting, cell adhesion, and cell spreading assays. Lastly, Y15 [a specific inhibitor of focal adhesion kinase (FAK)] was used to examine its potential to inhibit tumor growth in ESCA cells. Results: SPP1 was upregulated in ESCA tissues compared to the adjacent nontumorous tissues, which was closely associated with clinical stage, lymph node metastasis, histological subtype, and p53 mutation. A high expression of SPP1 indicated a poor clinical prognosis in patients with ESCA. The knockdown of SPP1 inhibited cell proliferative, migratory, and invasive capacities in ESCA cells. GSEA indicated that the focal adhesion pathway was closely related with SPP1 in ESCA. Further studies confirmed that the knockdown of SPP1 suppressed cell adhesion ability and reduced the expression of p-FAK and p-Erk in ESCA cells. In addition, Y15 inhibited FAK autophosphorylation and dramatically inhibited cell proliferation, migration, and invasion in ESCA cells. Conclusions: SPP1 promotes tumor progression in ESCA by activating FAK/Erk pathway, and FAK is a potential therapeutic target to overcome tumor recurrence and metastasis of ESCA.

Actin filament-associated protein 1-antisense RNA1 promotes the development and invasion of tongue squamous cell carcinoma via the AFAP1-AS1/miR-133a-5p/ZIC2 axis.

BACKGROUND: The present study aimed to explore the biological role and underlying mechanism of the long non-coding RNA actin filament-associated protein 1-antisense RNA1 (lncRNA AFAP1-AS1) in the progression of tongue squamous cell carcinoma (TSCC). METHODS: A quantitative reverse transcriptase-PCR (RT-qPCR) was conducted to assess relative levels of the miR-133a-5p, lncRNAs AFAP1-AS1 and zinc finger family member 2 (ZIC2) in TSCC cell lines and specimens, whereas ZIC2 protein levels were measured using western blotting. After modifying the levels of expression of lncRNA AFP1-AS1, miR-133a-5p and ZIC2 using lentivirus or plasmid transfection, we examined AKT/epithelial-mesenchymal transition signaling pathway alterations, in vivo carcinogenesis of TSCC in nude mice and in vitro malignant phenotypes. A dual-luciferase reporter assay was conducted to confirm the targeting relationship between ZIC2 and miR-133a-5p, as well as between miR-133a-5p and lncRNA AFAP1-AS1. Based on The Cancer Genome Atlas (TCGA) database, we additionally validated AFP1-AS1. The potential biological pathway for AFP1-AS1 was investigated using gene set enrichment analysis (GSEA). We also evaluated the clinical diagnostic capacities of AFP1-AS1 and clustered the most potential biomarkers with the Mfuzz expression pattern. Finally, we also made relevant drug predictions for AFP1-AS1. RESULTS: In TSCC cell lines and specimens, lncRNA AFAP1-AS1 was upregulated. ZIC2 was upregulated in TSCC cells as a result of lncRNA AFAP1-AS1 overexpression, which also promoted TSCC cell migration, invasion, viability, and proliferation. Via the microRNA sponge effect, it was found that lncRNA AFAP1-AS1 could upregulate ZIC2 by competitively inhibiting miR-133a-5p. Interestingly, knockdown of ZIC2 reversed the biological roles of lncRNA AFAP1-AS1 with respect to inducing malignant phenotypes in TSCC cells. In addition, in vivo overexpression of lncRNA AFAP1-AS1 triggered subcutaneous tumor growth in nude mice implanted with TSCC cells and upregulated ZIC2 in the tumors. The TCGA database findings revealed that AFAP1-AS1 was significantly upregulated in TSCC specimens and had good clinical diagnostic value. The results of GSEA showed that peroxisome proliferator-activated receptor signaling pathway was significantly correlated with low expression of AFP1-AS1. Finally, the results of drug prediction indicated that the group with high AFAP1-AS1 expression was more sensitive to docetaxel, AZD4547, AZD7762 and nilotinib. CONCLUSIONS: The upregulation of lncRNA AFAP1-AS1, which increases TSCC cell viability, migration, proliferation and invasion via the AFAP1-AS1/miR-133a-5p/ZIC2 axis, aids in the progression of TSCC.

Induction of local immunosuppression in allogeneic cell transplantation by cell-type-specific expression of PD-L1 and CTLA4Ig.

Immune rejection has long hindered allogeneic cell transplantation therapy. Current genetic modification approaches, including direct targeting of major histocompatibility complex or constitutive expression of immune inhibitory molecules, exhibit drawbacks such as severe adverse effects or elevated tumorigenesis risks. To overcome these limitations, we introduce an innovative approach to induce cell-type-specific immune tolerance in differentiated cells. By engineering human embryonic stem cells, we ensure the exclusive production of the immune inhibitory molecules PD-L1/CTLA4Ig in differentiated cells. Using this strategy, we generated hepatocyte-like cells expressing PD-L1 and CTLA4Ig, which effectively induced local immunotolerance. This approach was evaluated in a humanized mouse model that mimics the human immune system dynamics. We thus demonstrate a robust and selective induction of immunotolerance specific to hepatocytes, improving graft survival without observed tumorigenesis. This precise immune tolerance strategy holds great promise for advancing the development of stem cell-based therapeutics in regenerative medicine.

Effects of abnormal expression of CD73 on malignant phenotype of nasopharyngeal carcinoma.

Cluster of differentiation (CD) 73, encoded by the NT5E gene, plays important enzymatic and non-enzymatic roles in cells. There is growing evidence show that CD73 is a key regulator in the development of tumor. Nasopharyngeal carcinoma (NPC) is one of the most common cancers in east and southeast Asia. It is urgent to know more about the mechanism of NPC development and find diagnostic markers for the patients. In this research, we carried out western blot, immunohistochemistry and qRT-PCR to investigate the expression level of CD73 and found that NPC tissues had higher level of CD73 than normal tissues. We also detected the relationship between its expression level with the clinicopathological features and prognosis of NPC patients. The results showed that CD73 expression was related to the clinical stages, lymph node metastasis and survival state of NPC patients. More importantly, patients with higher expression of CD73 had poorer prognosis. Then, CD73 was knocked down in NPC cells (CNE2 and CNE1), and its effects on cell proliferation and migration were investigated by CCK8, colony formation, Transwell and wound-healing assays. We found that knocking down the expression of CD73 in NPC cells could inhibit cells malignant phenotype. Collectively, CD73 plays important roles in NPC malignant behavior and might act as a novel target for the diagnosis and treatment of NPC.

A novel frequency-dependent lattice Boltzmann model with a single force term for electromagnetic wave propagation in dispersive media.

Electromagnetic wave simulation is of pivotal importance in the design and implementation of photonic nano-structures. In this study, we developed a lattice Boltzmann model with a single extended force term (LBM-SEF) to simulate the propagation of electromagnetic waves in dispersive media. By reconstructing the solution of the macroscopic Maxwell equations using the lattice Boltzmann equation, the final form only involves an equilibrium term and a non-equilibrium force term. The two terms are evaluated using the macroscopic electromagnetic variables and the dispersive effect, respectively. The LBM-SEF scheme is capable of directly tracking the evolution of macroscopic electromagnetic variables, leading to lower virtual memory requirement and facilitating the implementation of physical boundary conditions. The mathematical consistency of the LBM-SEF with the Maxwell equations was validated by using the Champman-Enskog expansion; while three practical models were used to benchmark the numerical accuracy, stability, and flexibility of the proposed method.

PCK2 inhibits lung adenocarcinoma tumor cell immune escape through oxidative stress-induced senescence as a potential therapeutic target.

Background: Our research aimed to better understand how phosphoenolpyruvate carboxykinase 2 (PCK2) is linked to survival outcomes in lung cancer patients. Methods: We confirmed PCK2 expression and its association with the outcome of lung cancer patients using The Cancer Genome Atlas (TCGA) database. PCK2 and immune cell connections were investigated using data from the Tumor IMmune Estimation Resource (TIMER) and TCGA repositories. We used the CancerSEA database to examine the links between PCK2 expression and the efficiency of lung adenocarcinomas, and a T-distributed Stochastic Neighbor Embedding (T-SNE) map was constructed to show the expression profile of PCK2 in single cells in TCGA lung adenocarcinoma samples. The potential mechanism of action was finally investigated using Gene Set Enrichment Analysis (GSEA) enrichment analysis, Gene Ontology (GO) pathway enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Results: The expression of PCK was lower in lung adenocarcinoma tumor tissues than in paracancerous tissues. Patients with lung adenocarcinoma who expressed PCK2 at high levels fared better in overall survival (OS), disease-specific survival (DSS), and progression free interval (PFI). PCK2 was positively correlated with programmed cell death 1 (PDCD1) expression, and its mutation rate in lung adenocarcinoma was 0.53%. CancerSEA research revealed that in lung adenocarcinoma, PCK2 was negatively correlated with epithelial-mesenchymal transition (EMT) and hypoxia. Gene ontology and KEGG enrichment analysis revealed PCK2-coexpressed genes influenced the onset and progression of lung adenocarcinoma by modulating the activity of DNA-binding transcriptional activators, the specificity of RNA polymerase II, the interaction between neuroactive ligands and their receptors, and the cAMP signaling pathway. The prognosis for lung adenocarcinoma was shown to vary according to whether PCK2 was involved in the response to oxidative stress-induced senescence, gene silencing, cell cycle, and other biological processes. Conclusions: An increased expression of PCK2 may be employed as a novel prognostic biomarker in patients with lung adenocarcinoma and has been shown to increase OS, DSS, and PFI. Improving the prognosis of lung adenocarcinoma by interference with PCK2 may be possible since it induces senescence through the oxidative stress response and blocks the immune escape of tumor cells. These results point to a probable target anticancer treatment development in lung adenocarcinoma.

Lipopolysaccharide Preconditioning Restricts Microglial Overactivation and Alleviates Inflammation-Induced Depressive-like Behavior in Mice.

Overactive microglia and severe neuroinflammation play crucial roles in the development of major depressive disorder. Preconditioning with lipopolysaccharide (LPS) provides protection against severe neuroinflammation. However, administering high doses of LPS to mice triggers depressive symptoms. Therefore, the optimal dose of LPS preconditioning needs to be determined by further experiments. LPS preconditioning is an effective agent in anti-inflammation and neuroprotection, but the mechanism by which LPS preconditioning acts in depression remain unclear. This study finds that the anti-inflammation mechanism of low-dose LPS preconditioning is mainly dependent on G-protein-coupled receptor 84 (GPR84). We use low-dose LPS for preconditioning and re-challenged mice or BV2 microglia with high-dose LPS. In addition, RNA-seq is used to explore underlying changes with LPS preconditioning. Low-dose LPS preconditioning reduces the expression of pro-inflammatory mediators and inhibits microglial activation, as well as suppresses the depressive-like behavior when the mice are re-challenged with high-dose LPS. Further investigation reveals that the tolerance-like response in microglia is dependent on the GPR84. Here, we show that low-dose LPS preconditioning can exert anti-inflammation effects and alleviates inflammation-induced depressive-like behavior in mice. As a potential therapeutic target for depression, LPS preconditioning needs to be given further attention regarding its effectiveness and safety.

Generation of human otic neuronal organoids using pluripotent stem cells.

Otic neurons, also known as spiral ganglion neurons (SGNs) in mammalian cochlea, transmit electrical signals from sensory hair cells to cochlear nuclei of the auditory system. SGNs are sensitive to toxic insults, vulnerable to get irreversible damaged and hardly regenerate after damage, causing persistent sensorineural hearing loss. Yet, to get authentic SGNs for research or therapeutic purpose remains challenging. Here we developed a protocol to generate human otic neuronal organoids (hONOs) from human pluripotent stem cells (hESCs), in which hESCs were step-wisely induced to SGNs of the corresponding stages according to their developmental trajectory. The hONOs were enriched for SGN-like cells at early stage, and for both neurons and astrocytes, Schwann cells or supporting cells thereafter. In these hONOs, we also determined the existence of typical Type I and Type II SGNs. Mature hONOs (at differentiation Day 60) formed neural network, featured by giant depolarizing potential (GDP)-like events and rosette-organized regions-elicited calcium traces. Electrophysiological analysis confirmed the existence of glutamate-responsive neurons in these hONOs. The otic neuronal organoids generated in this study provide an ideal model to study SGNs and related disorders, facilitating therapeutic development for sensorineural hearing loss.

Twin-Twin Transfusion Syndrome After Radiofrequency Ablation in Dizygotic Monochorionic Triamniotic Triplet Pregnancy With Sex Discordance: A Case Report.

Background: We report a case of dizygotic monochorionic triamniotic triplet pregnancy. Twin-twin transfusion syndrome was subsequently diagnosed combined with sex discordance in the two surviving fetuses after one fetus was reduced, which is extremely rare and has not been previously reported. Case Presentation: After reducing one fetus by radiofrequency ablation of a monochorionic triamniotic triplet pregnancy, twin-twin transfusion syndrome was subsequently diagnosed combined with sex discordance in the two surviving fetuses. Amniotic fluid for chromosome analysis showed normal karyotype 46, XY/46, XX of the donor and recipient fetus, and short tandem repeat (STR) analysis revealed dizygotic twins. Conclusions: Through this is an unusual case, we aim to emphasize the importance of accurate diagnosis of chorionicity and zygosity in sex discordant triplet pregnancy, which is the key to appropriate clinical management.

Continual Deletion of Spinal Microglia Reforms Astrocyte Scar Favoring Axonal Regeneration.

Astrocyte scar formation after spinal cord injury (SCI) efficiently limits the accurate damage but physically restricts the following axon regeneration. Lately, fine tuning scar formation is becoming a novel strategy to develop SCI treatment, yet how to leverage these opposite effects remains challenging. Here, utilizing an improved drug administration approach, we show that in a mouse model of spinal cord injury, continual deletion of microglia, especially upon scar formation, by pexidartinib decreases the amount of microglia-derived collagen I and reforms the astrocyte scar. The astrocytes become less compacted in the scar, which permits axon regeneration and extension. Although continual microglia deletion did not significantly improve the locomotive performance of the SCI mice, it did ameliorate their weight loss, possibly by improving their relevant health conditions. We thus identified a novel approach to regulate astrocyte scars for improved axon regeneration, which is indicative of the clinical treatment of SCI patients.

[Application of modified V-Y advancement flap for reconstruction of facial defect].

Objective:To investigate the clinical effect of modified V-Y advancement flap for reconstruction of facial skin defect. Methods:Thirty-eight patients with facial skin tumors underwent individual tumor resection according to pathological type and lesion depth. Based on the defect site and size, appropriate V-Y advancement flap was designed to reconstruct the skin defect in one stage. There were 9 cases of classic subcutaneous tissue pedicle V-Y advancement flap, 24 cases of modified subcutaneous tissue pedicle V-Y advancement flap and 5 cases of perforated V-Y advancement flap in our study. Results:Among the 38 patients, 34 cases had primary healing. Two cases developed necrosis at the edge of the flap and healed after debridement. Local infection occurred in 2 cases, which healed after short-term dressing change. Postoperative mild eyelid ectropion occurred in 2 cases and oral horn displacement in 1 case. The patients were followed up for 6-36 months postoperatively, and the function and appearance recovered well. One case had local recurrence and 3 cases had parotid lymph node metastasis, which were removed again and supplemented with radiotherapy. Conclusion:The improved design of V-Y advancement flap can enlarge the scope of facial defect reconstruction, and achieve good appearance and function.

Flexoelectricity Driven Fano Resonance in Slotted Carbon Nanotubes for Decoupled Multifunctional Sensing.

Multifunctionality, interference-free signal readout, and quantum effect are important considerations for flexible sensors equipped within a single unit towards further miniaturization. To address these criteria, we present the slotted carbon nanotube (CNT) junction features tunable Fano resonance driven by flexoelectricity, which could serve as an ideal multimodal sensory receptor. Based on extensive ab initio calculations, we find that the effective Fano factor can be used as a temperature-insensitive extrinsic variable for sensing the bending strain, and the Seebeck coefficient can be used as a strain-insensitive intrinsic variable for detecting temperature. Thus, this dual-parameter permits simultaneous sensing of temperature and strain without signal interference. We further demonstrate the applicability of this slotted junction to ultrasensitive chemical sensing which enables precise determination of donor-type, acceptor-type, and inert molecules. This is due to the enhancement or counterbalance between flexoelectric and chemical gating. Flexoelectric gating would preserve the electron-hole symmetry of the slotted junction whereas chemical gating would break it. As a proof-of-concept demonstration, the slotted CNT junction provides an excellent quantum platform for the development of multistimuli sensation in artificial intelligence at the molecular scale.

Effects of Water Immersion and Acidification on Nano-Pores in Tight Sandstones-A Case Study of the Gaotaizi Reservoir, Songliao Basin.

Hydraulic fracturing and acidification are among the most commonly used methods for stimulating the tight oil reservoirs and improving oil recovery. Therefore, examining the effects of water immersion and acidification on tight oil reservoirs is important for oilfield development plans. Core flooding testing, which analyzes the influence of core permeability variations before and after acid injection on the reservoir quality, is the conventional research method; however, it is difficult to observe the changes in minerals and pores caused by acidulation and water immersion in situ. In this study, we conduct field-emission scanning electron microscopy (FE-SEM), MAPS, the quantitative evaluation of minerals through scanning electronic microscopy (QEM-SCAN), and describe the types of pores in tight sandstone. Further, the effects of water immersion and acidification on pores in tight sandstone were studied. The results indicate that: (1) intergranular pores, intragranular dissolution pores, clay mineral intercrystalline pores, and micro-cracks were developed in the Gaotaizi tight sandstone in Songliao Basin, with the intergranular pores observed to be dominant; (2) the hydration of clay minerals induced by water injection caused plugging of pores at the nanometer- micrometer scale, and plane porosity is slightly reduced (˜0.86%); (3) acidification resulted in the dissolution of carbonate minerals, increasing the porosity of the reservoir, therefore, the increase in porosity is influenced by the carbonate mineral content. We recommend that future studies should investigate the content, type, and distribution of carbonate minerals in the operation area. During the process of reservoir stimulation, such as acidification and CO2 injection- and-production, the influence of carbonate minerals dissolution on oil production should be considered.

Exosomal lncRNA DOCK9-AS2 derived from cancer stem cell-like cells activated Wnt/ß-catenin pathway to aggravate stemness, proliferation, migration, and invasion in papillary thyroid carcinoma.

Exosomal long non-coding RNAs (lncRNAs) are crucial factors that mediate the extracellular communication in tumor microenvironment. DOCK9 antisense RNA2 (DOCK9-AS2) is an exosomal lncRNA which has not been investigated in papillary thyroid carcinoma (PTC). Based on the result of differentially expressed lncRNAs in PTC via bioinformatics databases, we discovered that DOCK9-AS2 was upregulated in PTC, and presented elevation in plasma exosomes of PTC patients. Functionally, DOCK9-AS2 knockdown reduced proliferation, migration, invasion, epithelial-to-mesenchymal (EMT) and stemness in PTC cells. PTC-CSCs transmitted exosomal DOCK9-AS2 to improve stemness of PTC cells. Mechanistically, DOCK9-AS2 interacted with SP1 to induce catenin beta 1 (CTNNB1) transcription and sponged microRNA-1972 (miR-1972) to upregulate CTNNB1, thereby activating Wnt/ß-catenin pathway in PTC cells. In conclusion, PTC-CSCs-derived exosomal lncRNA DOCK9-AS2 activated Wnt/ß-catenin pathway to aggravate PTC progression, indicating that DOCK9-AS2 was a potential target for therapies in PTC.

Docosahexaenoic acid inhibits Ca2+ influx and downregulates CaSR by upregulating microRNA-16 in pulmonary artery smooth muscle cells.

Docosahexaenoic acid (DHA) is reported to have the potential to ameliorate pulmonary arterial hypertension (PAH), while the specific mechanism is still obscure. This study aims to investigate the function of DHA in pulmonary artery smooth muscle cells (PASMCs) and explore the underlying mechanism. In our study, DHA was used to incubate PASMCs. Cytosolic-free Ca2+ concentration ([Ca2+ ]cyt) was measured using Fluo-3 AM method. Real-time polymerase chain reaction was used to detect microRNA-16 (miR-16) and calcium-sensing receptor (CaSR) messenger RNA expression levels. CCK-8 assay, BrdU assay, and Transwell assay were employed to detect the effects of DHA on proliferation and migration of PASMCs. CaSR was confirmed as a direct target of miR-16 using dual-luciferase assay, polymerase chain reaction, and Western blot analysis. It was found that DHA significantly inhibited PASMC proliferation and migration and decreased [Ca2+ ]cyt. After transfection of miR-16 mimics, proliferation and migration ability of PASMCs were significantly inhibited, whereas opposite effects were observed after miR-16 inhibition. [Ca2+ ]cyt was also inhibited by miR-16 transfection. DHA then promoted the expression of miR-16, and the effects of DHA on PASMCs were annulled when miR-16 was inhibited. CaSR was identified as a direct target of miR-16. CaSR was inhibited directly by miR-16 and indirectly by DHA. In conclusion, DHA inhibits the proliferation and migration of PASMCs, and probably ameliorates PAH via regulating miR-16/CaSR axis.

Elevated O-GlcNAcylation Promotes Malignant Phenotypes of Hypopharyngeal Squamous Cell Carcinoma by Stabilizing Nrf2 through Regulation of the PI3K/Akt Pathway.

BACKGROUND AND PURPOSE: O-GlcNAcylation is a significant protein posttranslational modification with O-linked ß-N-acetylglucosamine (GlcNAc) for intracellular signaling. Elevated O-GlcNAcylation contributes to cell proliferation, cell migration, cell apoptosis and signal transduction in various cancers. However, the expression level and functional role of O-GlcNAcylation in Hypopharyngeal Squamous Cell Carcinoma (HSCC) is not clearly elucidated. Nuclear factor erythroid-2-related factor 2 (Nrf2) is a master transcriptional factor that has been found to be aberrantly activated in HSCC. Here, we provide a molecular rationale between O-GlcNAcylation and Nrf2 in HSCC patients. METHODS: The protein levels of O-GlcNAcylation and Nrf2 in HSCC tissues were detected by immunohistochemistry technique and western blot analysis. Then, O-GlcNAcylation knockdown HSCC cells were applied in this study. Cell proliferation was detected by CCK8, colony-forming analysis, and cell cycle assays. Cell migration and invasion ability was evaluated by transwell assays. Cell apoptosis was measured by TUNEL analysis. RESULTS: O-GlcNAcylation was obviously up-regulated in HSCC tissues, which correlated with tumor size and lymph node metastasis. In addition, the protein level of Nrf2 was found to positively correlate with the expression of O-GlcNAcylation both in vivo and in vitro. Knockdown of O-GlcNAcylation significantly inhibited HSCC cell growth, suppressed cell migration, and promoted cell apoptosis, whereas overexpression of Nrf2 reversed these phenotypes. Mechanismly, the upregulation of O-GlcNAcylation promoted the phosphorylation of Akt, leading to the stabilization of Nrf2; this could be attenuated by inhibition of the PI3K/Akt signaling pathway. CONCLUSION: Here, we provide a molecular association between O-GlcNAcylation and Nrf2 in HSCC patients, thus providing valuable therapeutic targets for the disease.

Para-aortic lymphadenectomy did not improve overall survival among women with type I endometrial cancer.

OBJECTIVE: To compare outcomes and prognosis among women with type I endometrial cancer undergoing hysterectomy and bilateral salpingo-oophorectomy (H-BSO) with or without systematic pelvic lymphadenectomy (PLD) or para-aortic lymphadenectomy (PALD). METHODS: Retrospective review of women postoperatively diagnosed with type I endometrial cancer who underwent H-BSO at a university hospital in Chengdu, China (January 2010 to June 2012). Women were divided into no lymphadenectomy (PLD-/PALD-), systematic pelvic lymphadenectomy (PLD+/PALD-), or combined pelvic and para-aortic lymphadenectomy (PLD+/PALD+) groups. Follow-up was by telephone. Postoperative outcomes and prognosis were compared and risk factors were analyzed. RESULTS: In total, 333 women met the inclusion criteria: 121 underwent PLD+/PALD-, 166 underwent PLD+/PALD+, and 46 underwent PLD-/PALD-. There were no differences in pre-operative characteristics among the groups (all P>0.05). The PLD+/PALD+ group had a higher laparotomy rate (P=0.001), the PLD-/PALD- group had shorter operation time (P=0.001) and lower blood loss (P<0.001). There were no differences between the PLD+/PALD- and PLD+/PALD+ groups. Overall, 291 women had sufficient follow-up data; there was no difference in overall survival, and PALD was not a predictor of survival. CONCLUSION: Postoperative outcomes were similar among all surgical groups; a survival benefit of PALD was not demonstrated.

Transcriptome analysis of tongue cancer based on high­throughput sequencing.

Tongue cancer is one of the most common types of cancer, but its molecular etiology and pathogenesis remain unclear. The aim of the present study was to elucidate the pathogenesis of tongue cancer and investigate novel potential diagnostic and therapeutic targets. Four matched pairs of tongue cancer and paracancerous tissues were collected for RNA sequencing (RNA­Seq), and the differentially expressed genes were analyzed. The RNA­Seq data of tongue cancer tissues were further analyzed using bioinformatics and reverse transcription­quantitative PCR analysis. The sequenced reads were quantified and qualified in accordance with the analysis demands. The transcriptomes of the tongue cancer tissues and paired paracancerous tissues were analyzed, and 1,700 upregulated and 2,249 downregulated genes were identified. Gene Ontology analysis uncovered a significant enrichment in the terms associated with extracellular matrix (ECM) organization, cell adhesion and collagen catabolic processes. Kyoto Encyclopedia of Genes and Genomes analysis demonstrated that these differentially expressed genes were mainly enriched in the focal adhesion pathway, ECM­receptor interaction pathway, phosphoinositide 3­kinase (PI3K)­Akt pathway, and cell adhesion molecules. Comprehensive analyses of the gene tree and pathway network revealed that the majority of cell cycle genes were upregulated, while the majority of the genes associated with intracellular response, cell adhesion and cell differentiation were downregulated. The ECM­receptor interaction, focal adhesion kinase (FAK) and PI3K­Akt pathways were closely associated with one another and held key positions in differential signaling pathways. The ECM­receptor, FAK and PI3K­Akt signaling pathways were found to synergistically promote tongue cancer occurrence and progression, and may serve as potential diagnostic and therapeutic targets for this type of cancer.

MicroRNA-143-5p modulates pulmonary artery smooth muscle cells functions in hypoxic pulmonary hypertension through targeting HIF-1α.

This paper explores the potential mechanism of microRNA-143-5p regulation effects on pulmonary artery smooth muscle cells (PASMCs) functions in hypoxic pulmonary hypertension (HPH) via targeting HIF-1a, which may offer a new idea for HPH therapy. PASMCs were transfected with mimics control/miR-143-5p mimics or inhibitor control/miR-143-5p inhibitor. We used Western blotting and RT-qPCR to detect the protein and mRNA expressions, CCK-8 assay to detect cellular viability, Annexin V-FITC/PI staining and caspase- 3/cleaved caspase-3 protein to evaluate cellular apoptosis, transwell migration experiment for cellular migration measurement and Dual luciferase reporter gene assay to prove the target of miR-143-5p. Cells under hypoxic condition presented the decreased protein and mRNA expressions of α-smooth muscle actin (SM-α-actin), Myocardin, smooth muscle myosin heavy chain (SMMHC), and smooth muscle-22α (SM22α), Calponin1 and Hypoxia-inducible factor-1α(HIF-1α), the increased cell viability and miR-143-5p level; Overexpression of miR-143-5p obviously reduced vascular smooth muscle-specific contraction marker protein levels and cellular apoptosis, increased cellular migration of PASMCs with hypoxia stimulation; Low-expression of miR-143-5p caused the opposite changes, while co-transfected with Si HIF-1 α blocked the beneficial effects of miR-143-5p inhibition on PASMCs under hypoxia. MicroRNA-143-5p can promote the phenotype conversion, proliferation and migration of pulmonary artery smooth muscle cells under hypoxic condition through direct targeting of HIF-1α.

Inhibition of JNK ameliorates depressive-like behaviors and reduces the activation of pro-inflammatory cytokines and the phosphorylation of glucocorticoid receptors at serine 246 induced by neuroinflammation.

Depression is associated with immune dysregulation and the aberrant activity of the hypothalamic-pituitary-adrenal (HPA) axis. However, the neurobiological molecular mechanisms underlying these associations remain unclear. c-Jun amino-terminal kinase (JNK), an important modulator in inflammation and stress responses, is often critically implicated in the development of central nervous system diseases. However, whether and how JNK mediates neuroinflammation-induced depression remains largely unknown. In this study, we investigated the role of JNK in depressive-like behaviors induced by central lipopolysaccharide (LPS) infusion. The results showed that LPS infusion led to depressive-like behaviors, accompanied by increased proinflammatory cytokine expression, increased JNK activation, and upregulated glucocorticoid receptor (GR) phosphorylation at serine 246 (pGR-Ser246) in the habenula (Hb), amygdala (Amyg) and medial prefrontal cortex (mPFC). Treatment with SP600125, a known JNK inhibitor, prevented the LPS-induced hyper-activation of JNK and alleviated depressive-like behaviors. Moreover, LPS-induced increases in the expression levels of TNF-α, IL-1ß and pGR-Ser246 in these brain regions were reduced when the rats were treated with SP600125. Our results show, for the first time, that JNK activities in the Hb, Amyg, and mPFC are involved in the modulation of neuroinflammation-induced depression and participate in the regulation of the expression of proinflammatory cytokines and GR phosphorylation, which are pathological factors associated with depression. Our findings provide new insights into the mechanism of neuroinflammation-associated depression and suggest that the JNK pathway may be a potential target for treating inflammation-related depression.