Identification of prognostic m6A modification patterns and score system in melanoma patients.

N6-methyladenosine (m6A) is the most common modification on RNAs and LncRNAs. It plays an important role in cancer stem cell differentiation, T cell differentiation, and immune homeostasis. In this study, we explored the potential roles of m6A modification of RNA in melanoma and investigated the immune cell infiltration in tumor microenvironment in diverse m6Aclusters and different m6Ascore groups. A consensus clustering algorithm determined m6A modification patterns based on 14 m6A regulators, and further explored the biological functions and the connection with TME. An m6A-related gene signature (m6Ascore) was constructed based on m6A-related genes using principal component analysis. Three m6A modification patterns were identified based on 14 m6A regulators, named as m6Aclusters A-C. The prognosis of m6Acluster A was more favorable than m6Aclusters B and C, and it was more closely associated with immune regulation. To quantify the m6A modification patterns of individual tumor, an m6Ascore was constructed, and patients were classified into high and low m6Ascore groups. The low m6Ascore group, which had a favorable prognosis, was more relevant to immunology. The expression of PD-L1 was higher and the immunophenoscore (IPS) revealed stronger response to immunotherapy in the low m6Ascore group. This study identified 3 m6A modification patterns with different immune characteristics and constructed an m6Ascore system to predict prognosis and immunogenicity of patients, which is conducive to clinical prognosis judgment and individual treatment.

Glutathione safeguards TET-dependent DNA demethylation and is critical for the acquisition of totipotency and pluripotency during preimplantation development.

During early development, both genome-wide epigenetic reprogramming and metabolic remodeling are hallmark changes of normal embryogenesis. However, little is known about their relationship and developmental functions during the preimplantation window, which is essential for the acquisition of totipotency and pluripotency. Herein, we reported that glutathione (GSH), a ubiquitous intracellular protective antioxidant that maintains mitochondrial function and redox homeostasis, plays a critical role in safeguarding postfertilization DNA demethylation and is essential for establishing developmental potential in preimplantation embryos. By profiling mitochondria-related transcriptome that coupled with different pluripotency, we found GSH is a potential marker that is tightly correlated with full pluripotency, and its beneficial effect on prompting developmental potential was functionally conformed using in vitro fertilized mouse and bovine embryos as the model. Mechanistic study based on preimplantation embryos and embryonic stem cells further revealed that GSH prompts the acquisition of totipotency and pluripotency by facilitating ten-eleven-translocation (TET)-dependent DNA demethylation, and ascorbic acid (AsA)-GSH cycle is implicated in the process. In addition, we also reported that GSH serves as an oviductal paracrine factor that supports development potential of preimplantation embryos. Thus, our results not only advance the current knowledge of functional links between epigenetic reprogramming and metabolic remodeling during preimplantation development but also provided a promising approach for improving current in vitro culture system for assisted reproductive technology.

Preparation of layering-structured magnetic fluorescent liposomes and labeling of HepG2 cells.

BACKGROUND: At present, surgical resection and chemotherapy are still the main treatments for hepatocellular carcinoma and other cancers, but the curative effect and survival rate are not ideal. OBJECTIVE: In this study, we aim to prepare a carrier with low toxicity, high biocompatibility and targeted transport for the treatment of hepatocellular carcinoma. METHODS: CdSe quantum dots (QDs) modified with oleic acid were synthesized. Then hydrophobic CdSe QDs and hydrophilic super-paramagnetic Fe3O4 particles were encapsulated into different layers of liposomes to form magnetic fluorescent liposomes (MFLs). MFLs in the aqueous would quickly drift towards the external magnet and the entire process was clearly observed with fluorescence microscope. The fluorescence spectra revealed that the fluorescence properties of MFLs were similar to that of CdSe QDs. RESULTS: QDs had an average size of 3.32 nm with good fluorescence properties. The size of MFLs was about 100 nm (transmission electron microscopy (TEM) analysis showed the average size of MFLs was about 82.8 nm and dynamic light scattering (DLS) detection showed 111.9 nm). After being cultured with MFLs for 8 h, HepG2 cells were labeled by MFLs, and good fluorescence images were obtained. MTT analysis also expressed their good biocompatibility. CONCLUSION: The prepared MFLs had multi-function and could be used as ideal drug carriers.

Corrigendum: A Prognostic Model Based on Immune-Related Long Non-Coding RNAs for Patients With Cervical Cancer.

[This corrects the article DOI: 10.3389/fphar.2020.585255.].

Prospective Trial of Near-Infrared Spectroscopy for Continuous Noninvasive Monitoring of Free Fibular Flaps.

INTRODUCTION: Some free flaps develop postoperative vessel thrombosis, which influences the flap survival rate. Early discovery and identification of vascular crisis are critical to the success rate of flap salvage. The primary aims of this study were to determine the features of postoperative blood supply changes in fibular flaps with normal and abnormal blood flow, using near-infrared spectroscopy (NIRS), to monitor oxygenation and blood flow, and to characterize the probable risk factors for vascular crisis. METHODS: Sixty-three consecutive patients undergoing reconstruction of unilateral mandibular defects with free fibular flaps at the Peking University School of Stomatology were included. Patients were divided into 2 groups, A (n = 38) and B (n = 25); fibular flaps in group A underwent continuous NIRS monitoring from immediately postoperatively until 7 days postoperatively (approximately 150 hours), whereas fibular flaps and opposite mandibles in group B underwent intermittent monitoring: once every 4 hours during the first 24 hours postoperatively and once every 12 hours from 24 to 168 hours postoperatively. RESULTS: Six fibular flaps developed vascular thromboses: 4 were venous thromboses and 2 were arterial thromboses; 5 were rescued after exploration. Of 6 regional oxygen saturation (rSO2) values in the continuous monitoring group, 4 showed no significant differences at any time point compared with the intermittent monitoring group (P > 0.05). The rSO2 of the fibular flap was significantly different from that in the opposite mandible in the first 36 hours postoperatively (P < 0.05). This difference decreased over time. During the initial period of venous thrombosis, rSO2, deoxyhemoglobin, and oxygenated hemoglobin all rose slightly, then showed simultaneous rapid reduction. However, the magnitude of reduction was smaller for deoxyhemoglobin than for oxygenated hemoglobin. CONCLUSIONS: Near-infrared spectroscopy can be used for noninvasive and reliable assessment of oxygenation and blood flow in free flaps through continuous, real-time monitoring. It is also portable, inexpensive, and simple to operate. In addition, the detection depth of NIRS is up to 2.0 cm, so it can be used to monitor buried flaps with depths <2.0 cm.

Assaying of Cu2+ with near-infrared l-cysteine-capped CdSeTe/CdS quantum dots and its effect on cell imaging.

Near-infrared (NIR) core-shell CdSeTe/CdS quantum dots (QDs) modified with l-cysteine were synthesized in aqueous solution. The QDs had a special NIR-emitting spectrum, high fluorescence stability and low cytotoxicity. In addition, they exhibited an obvious fluorescence quenching when Cu2+ was present. An NIR nanosensor was prepared for rapidly, sensitively, and selectively determining Cu2+ in solution quantitatively and monitoring the changes in Cu2+ in cells with fluorescence imaging in a semiquantitative way. The linear relationship between the relative fluorescence intensity (F0 /F) and the concentration of Cu2+ from 5.12 × 10-8  M to 2.56 × 10-5  M in solution was observed using an NIR fluorescence spectrophotometer with R2 equal to 0.9958. Moreover, in the experiment with the fluorescence microscope, F0 /F versus the concentration of Cu2+ from 5.00 × 10-8  M to 7.68 × 10-6  M also showed a good linear relationship with R2 equal to 0.9817. Practical water sample ion detecting experiments had good accuracy and recovery rates. Cell experiments showed that the NIR imaging intensity of cells was inversely proportional to the concentration of copper ions, therefore NIR QDs have great potential for detection of metal ions in solution and in cells.

A Prognostic Model Based on Immune-Related Long Non-Coding RNAs for Patients With Cervical Cancer.

Objectives: The study is performed to analyze the relationship between immune-related long non-coding RNAs (lncRNAs) and the prognosis of cervical cancer patients. We constructed a prognostic model and explored the immune characteristics of different risk groups. Methods: We downloaded the gene expression profiles and clinical data of 227 patients from The Cancer Genome Atlas database and extracted immune-related lncRNAs. Cox regression analysis was used to pick out the predictive lncRNAs. The risk score of each patient was calculated based on the expression level of lncRNAs and regression coefficient (ß), and a prognostic model was constructed. The overall survival (OS) of different risk groups was analyzed and compared by the Kaplan-Meier method. To analyze the distribution of immune-related genes in each group, principal component analysis and Gene set enrichment analysis were carried out. Estimation of STromal and Immune cells in MAlignant Tumors using Expression data was performed to explore the immune microenvironment. Results: Patients were divided into training set and validation set. Five immune-related lncRNAs (H1FX-AS1, AL441992.1, USP30-AS1, AP001527.2, and AL031123.2) were selected for the construction of the prognostic model. Patients in the training set were divided into high-risk group with longer OS and low-risk group with shorter OS (p = 0.004); meanwhile, similar result were found in validation set (p = 0.013), combination set (p < 0.001) and patients with different tumor stages. This model was further confirmed in 56 cervical cancer tissues by Q-PCR. The distribution of immune-related genes was significantly different in each group. In addition, the immune score and the programmed death-ligand 1 expression of the low-risk group was higher. Conclusions: The prognostic model based on immune-related lncRNAs could predict the prognosis and immune status of cervical cancer patients which is conducive to clinical prognosis judgment and individual treatment.

Real-time measurement of a probabilistic-shaped 200Gb/s DP-16QAM transceiver.

Continuous real-time measurements are demonstrated from a 200Gb/s format configurable CFP transceiver that uses dual-polarization probabilistic-shaped 16QAM (DP-PS16QAM) modulation. Placed in a 50GHz coherent DWDM transmission system, DP-PS16QAM achieves a back-to-back 1.8dB OSNR gain over uniform DP-16QAM. It also transports over 1940km with EDFA-only amplification, thus doubling propagation distance of uniform DP-16QAM. Furthermore, a 1Tb/s super-channel consisting of five 200Gb/s DP-PS16QAM sub-carriers is placed in a 200GHz grid, and it achieves over 1600km transmission and 5b/s/Hz SE with a raw SE at 6.86b/s/Hz.

USP10 regulates Musashi-2 stability via deubiquitination and promotes tumour proliferation in colon cancer.

Recent studies have demonstrated that ubiquitin-specific protease 10 (USP10) plays a catalytic role in tumour suppression mainly by deubiquitinating its target proteins to enhance their stabilities. However, we found that USP10 could interact with and regulate the expression of oncogenic factor Musashi-2 (MSI2). We investigated whether USP10 positively regulates the expression of MSI2 by deubiquitination and confirmed the type of polyubiquitin chain that is linked to MSI2. We also explored the role of USP10 in regulating the proliferation of colon cancer through different experiments. This study provides a completely new perspective in understanding the role of USP10 in deubiquitination. In the future, USP10 may serve as a target for colon cancer treatment.

Improvement of self-cleaning waterborne polyurethane-acrylate with cationic TiO2/reduced graphene oxide.

UV curable waterborne polyurethane acrylate (WPUA) with surfactant-modified TiO2/reduced graphene oxide (TiO2/rGO) nanocomposites were prepared and analyzed to improve their mechanical performance and self-cleaning ability. TiO2/rGO nanocomposites were prepared by a simple hydrothermal method with nano-TiO2 and graphene oxide, and modified with cationic surfactant (CTAB) to obtain a cationic TiO2/rGO (C-TiO2/rGO). Then, the obtained C-TiO2/rGO was incorporated into anionic waterborne polyurethane acrylate by in situ fabrication to obtain a composite emulsion (C-TiO2/rGO-WPUA). The results of Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM) showed that CTAB was successfully intercalated into TiO2/rGO, and TiO2 nanoparticles were evenly distributed on graphene sheets with good dispersibility. Compared to UV-cured neat WPUA and C-TiO2/rGO-WPUA, the mechanical properties and thermal stability of the composites were significantly improved. When the content of C-TiO2/rGO was 0.5%, the UV-cured composites had overall excellent performance. In particular, the WPUA composites exhibited good self-cleaning ability in photocatalysis. The photocatalytic degradation rate of methyl orange in 0.5% C-TiO2/rGO-WPUA reached 88.3% under 6 h visible light irradiation.

Cartilage oligomeric matrix protein is a prognostic factor and biomarker of colon cancer and promotes cell proliferation by activating the Akt pathway.

PURPOSE: Recent studies have determined that cartilage oligomeric matrix protein (COMP) plays a vital role in carcinogenesis. We sought to clarify the role of COMP in colon cancer. METHODS: We investigated gene expression data from The Cancer Genome Atlas (TCGA) dataset. Tissue microarrays (TMA) containing paired samples from 253 patients with colon cancer were subjected to immunostaining. COMP levels in serum of colon cancer patients and healthy donors were measured with ELISA. We established COMP-knockout cells using the CRISPR/Cas9 system and COMP-overexpressing cells using lentiviral vectors to detect the effects of COMP on colon cancer cells using Cell Counting Kit-8 (CCK8), colony formation, apoptosis detection kit, and tumorigenesis assays in nude mice. RESULTS: The analysis of TCGA dataset and the results of the TMA suggested that COMP expression levels were significantly higher in cancer tissues than in adjacent normal tissues. Moreover, high COMP expression was correlated with the poor outcome of colon cancer patients. COMP levels in the sera of preoperative patients with colon cancer were much higher than those in healthy donors and were significantly reduced after colectomy. Colon cancer cells without COMP were defective with respect to the ability to proliferate, colony formation, the ability to resist 5-Fluorouracil-induced apoptosis and the growth of xenograft tumors in mice. Contrasting results were observed in COMP overexpressed cells. COMP promoted colon cancer cell proliferation partially through the activation of PI3K/ Akt/ mTOR/ p70S6K pathway. CONCLUSIONS: COMP may be a novel prognostic indicator and biomarker and also a potential therapeutic target for colon cancer.

L-noradrenaline functionalized near-infrared fluorescence CdSeTe probe for the determination of urea and bioimaging of HepG2 Cells.

In this paper, near-infrared (NIR) light emitting L-noradrenaline functionalized CdSeTe QDs (NA-CdSeTe) were synthesized and applied to the biosensing of urea. When the pH value of NA-CdSeTe solution was adjusted from neutral to alkalinity, the noradrenaline on the surface of QDs would turn to quinone, which triggered the fluorescence quenching of NA-CdSeTe probe due to the charge transfer interactions between QDs and the proximal quinone. Based on the fact that the hydrolysis of urea in the presence of urease would release OH- and slightly change the pH value of the solution, the fluorescence intensity of NA-CdSeTe QDs could be linked to the enzymatic degradation of urea. The novel urea-biosensing system could effectively determinate urea in the dynamic concentration range from 0.057 to 13mmol/L. Furthermore, NA-CdSeTe probe could serve as an "optical window" for the bioimaging of urea with high selectivity in the serum samples and HepG2 cells. The urea-bioimaging system could effectively probe urea in the dynamic concentration range from 0.2 to 5mmol/L. This NIR probe was excellent candidate not only for its sensitive with urea but also its real-time bioimaging. We expected that this NIR probe based strategy could pave the way for developing simple, no enzyme immobilization required and good sensitive method related detections for further medical applications.

[A novel technique to preserve the alveolar ridge width following tooth extraction in the maxillary frontal area].

OBJECTIVE: To introduce a novel ridge preservation technique with micro-titanium plate avoiding the use of bone grafting materials, and evaluate the potential horizontal bone preservation effect of this new technique, applied on single maxillary central incisors after tooth extraction for future implant restoration. METHODS: Nine patients (six women and three men), mean age (26.0 ± 5.7) years(from 18 to 34 years) referred to the Department of Oral Implantology, Peking University School and Hospital of Stomotology, were selected and diagnosed with unsalvageable single middle incisor with fine general conditions, no signs of acute local inflammation, no ongoing or previous periodontitis, healthy neighboring teeth and intact buccal bone walls. Tooth extraction, delayed implant placement and implant-supported single crown restoration were selected as treatment plan. The teeth were extracted atraumatically with local anesthesia, followed by a reflection of a minor flap to exposed 2-3 mm of the buccal bone plate. After that, a micro-titanium plate was trimmed and bended to fit the convexity of the labial bone and fixed by two mini pins with intent to support the labial soft tissue. The flap was then repositioned over the micro titanium plate and secured with two single sutures. No bone grafting materials or releasing incisions were needed. The sockets were left to heal without any intention of primary wound closure. Cone-beam computed tomographic (CBCT) scans were obtained before and four months after tooth extraction. Horizontal ridge widths were measured with CBCT software, and the preservation effects were calculated and recorded by the percentage of horizontal ridge alteration. RESULTS: The nine extraction sockets were healed uneventfully. The average socket width before extraction was (7.51 ± 0.48) mm (6.92-7.82 mm). The average alveolar ridge labial-palatal width at the control point of the edentulous area was (6.81±0.44) mm (6.04-7.38 mm) 4 months after tooth extraction, the mean percentage of ridge width preserved was 90.87% ± 2.91% (87.28%-95.60%). CONCLUSION: This novel ridge preservation method by the usage of a micro-titanium plate did not interfere with the natural socket healing process, and at the same time, largely preserved the width of alveolar ridge without any bone grafting procedures. Long term results remain to be seen.

Evaluation of foot perfusion after fibula flap surgery.

Fibula flap is widely used in reconstruction work, whereas the low extremity blood supply alteration was unclear. This study would observe the blood oxygen saturation change of foot after harvesting fibula flap. The regional tissue oxygen saturation (rSO2) of the lateral side and inner side of feet was measured using near-infrared spectroscopy oxygen monitoring system (NIRS) before operation and in postoperative day 1 to 7, and the foot of the control side was measured as control. The rSO2 of the donor side foot decreased less than 5% (P < 0.05) in the first 8 hours after operation and recovered to the level of the control side later. NIRS is ideal for measuring rSO2 of donor side foot after fibula flap. The rSO2 of the donor side foot decreased less than 5% after fibula flap harvesting.

[Anatomic study of the hypoglossal nerve in hypoglossal-facial nerve anastomosis].

OBJECTIVE: To determine the optimal position of hypoglossal nerve in hypoglossal-facial nerve anastomosis and the eligibility of hypoglossal-facial nerve anastomosis with the cervical loop. METHODS: The cervical course and adjacent structures of the hypoglossal nerve were observed on 21 adult cadavers. The hypoglossal nerve and facial nerve were taken from 3 fresh specimens, and the number of the fasciculus and the cross-sectional area of the nerve were measured. RESULTS: The facial nerve trunk were monofascicular with a cross-sectional area of 5.1-/+0.2 (range 4.6-5.7) mm(2). The number of the fasciculus and the cross-sectional areas of the nerve trunk and the fasciculus were 1.6-/+0.8 (range 1-4) mm(2) , 7.5-/+0.7 mm(2) (range 6.8-8.0) mm(2), and 4.7-/+0.6 (4.1-5.5) mm(2), respectively, at the proximal segment of the hypoglossal nerve, 3.6-/+0.5 (1-5) mm(2) , 5.6-/+0.5 (4.9-6.1) mm(2) , and 1.6-/+0.4 (0.9-2.2) mm(2) at the distal segment, and 2.4-/+0.8 (1-3) mm(2), 1.1-/+0.7 (0.6-2.2) mm(2), and 0.5-/+0.3 (0.3-1.2) mm(2) at the cervical loop. CONCLUSION: The cervical loop is inadequate for facial nerve anastomosis and the proximal segment is large enough to allow partial harvesting of the hypoglossal nerve for neurotisation of the facial nerve.