Synovial fluid research based on SERS and SERRS for enhanced detection of biomarkers in staged osteoarthritis.

Surface-enhanced (resonance) Raman scattering (SER(R)S) can extremely enhance Raman intensity of samples, which is helpful for detecting synovial fluid (SF) that does not show Raman activity under normal conditions. In this study, SER(R)S spectra of SF from three different osteoarthritis (OA) stages were collected and analyzed for OA progress, finding that the content of collagen increased throughout the disease, while non-collagen proteins and polysaccharides decreased sharply at advanced OA stage accompanied by the increase of phospholipid. The spectral features and differences were enhanced by salting-out and centrifugation. Much more information on biomolecules at different OA stages was disclosed by using SERRS for the first time, these main trace components (ß-carotene, collagen, hyaluronic acid, nucleotide, and phospholipid) can be used as potential biomarkers. It indicates that SERRS has a more comprehensive ability to assist SERS in seeking micro(trace) biomolecules as biomarkers and facilitating accurate and efficient diagnosis and mechanism research of OA.

Long non-coding RNA HOTTIP exerts an oncogenic function by regulating HOXA13 in nasopharyngeal carcinoma.

BACKGROUND: The long non-coding RNA HOXA transcript at the distal tip (HOTTIP) and homeobox A13 (HOXA13) have been identified as oncogenes that play a pivotal role in tumorigenesis. However, their specific mechanisms of action in nasopharyngeal carcinoma (NPC) progression remain unclear. METHODS AND RESULTS: In the present study, RT-qPCR was employed to quantify RNA expression in NPC cells and tissues. Flow cytometry, MTT, CCK8 and colony formation assays were utilized to assess cell apoptosis and proliferation. Transwell assay was conducted to evaluate migration and invasion while Western blotting was performed for protein expression analysis. Our findings revealed that the expression of HOTTIP was significantly upregulated in NPC cell lines. Inhibition of HOTTIP could induce apoptosis and suppress proliferation, clonogenicity, invasion and metastasis in NPC cells. Knockdown of HOTTIP led to downregulation of HOXA13 expression, which subsequently inhibited the proliferation and metastasis in NPC cells. The inhibitory effects on cell proliferation and metastasis caused by HOTTIP silencing were rescued by HOXA13 overexpression. Additionally, there was a significant positive correlation between HOTTIP and HOXA13, which were found to be elevated in NPC tissues compared to normal tissues. CONCLUSIONS: We have determined that LncRNA HOTTIP facilitates tumorigenesis by modulating the expression of HOXA13 in NPC cells. Targeting HOTTIP/HOXA13 may be a promising therapeutic strategy for NPC.

Polysaccharides from Discarded Stems of Trollius chinensis Bunge Elicit Promising Potential in Cosmetic Industry: Characterization, Moisture Retention and Antioxidant Activity.

Unconventional polysaccharides as representative active substances from stems of Trollius chinensis Bunge (TC) were studied. Crude polysaccharides from the stems of TC (TCSP) and the petals of TC (TCPP) were extracted, and the moisture retention and antioxidation activities of both TCSP and TCPP in vitro were studied. The weight-average molar masses (Mw) of TCSP (6.07 × 105 Da) were lower than those of TCPP (9.72 × 105 Da). Glucuronic acid and xylose only existed in TCSP, and the molar ratio of galacturonic acid and mannose in TCSP was significantly higher than that in TCPP. No significant differences in moisture retention ability were found between TCSP and TCPP. The reducing capacity and dphenyl picryl hydrazinyl (DPPH) radical scavenging capacity of TCSP were slightly weaker than those of TCPP. The 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity of TCSP can be equivalent to that of TCPP. The moisture retention ability was not different between TCSP and TCPP, which are both highly homologous with traditional humectants. The antioxidation assays in vitro demonstrated that the antioxidant activity of TCSP is stronger compared to that of some plant-derived polysaccharides. The stems of TC can be a promising source of unconventional polysaccharides, which possess moisture retention and antioxidation capacities for the cosmetics industry.

Parapharyngeal Space Ganglioneuroma: Clinical Experience and Review of the Literature.

Ganglioneuroma is a rare benign tumor originating in the sympathetic ganglia, composed of differentiated ganglion cells, nerve sheath cells, and nerve fibers, which tend to occur in the posterior mediastinum, adrenal gland, retroperitoneal, and other locations, occurring in the head and neck is relatively rare, and parapharyngeal space involvement is extremely rare. In our report, we present 2 adult male patients whose preoperative imaging and fine needle cytology did not confirm the diagnosis of a parapharyngeal space mass and who completely resected the tumor through a combined cervical and oral approach. Finally, pathology confirmed ganglioneuroma; we also reviewed the English articles on parapharyngeal ganglioneuroma over the past 40 years, and summarized the diagnostic and treatment characteristics of parapharyngeal ganglioneuroma in combination with our cases to improve understanding of the disease.

Data reconstruction of daily MODIS chlorophyll-a concentration and spatio-temporal variations in the Northwestern Pacific.

Sea surface chlorophyll-a concentration (Chl-a) is a key proxy for phytoplankton biomass. Spatio-temporal continuous Chl-a data are important to understand the mechanisms of chlorophyll occurrence and development and track phytoplankton changes. However, the greatest challenge in utilizing daily Chl-a data is massive missing pixels due to orbital position and cloud coverage. This study proposes the application of a spatial filling method using the machine learning-based Extreme Gradient Boosting (BST) to reconstruct missing pixels of daily MODIS Chl-a data from 2007 to 2018. The approach is applied to different trophic biogeographical subregions of the Northwestern Pacific where it has complex phytoplankton dynamics and frequent data missing. Various environmental variables are taken into consideration, including meteorological forcing, geographic and topographic features, and oceanic physical components. The BST-reconstructed Chl-a (BST Chl-a) is validated using in-situ Chl-a measurements, VIIRS and Himawari-8 Chl-a products. The results show that the BST model is highly adaptive in reconstructing Chl-a data, and it performs well in pelagic, offshore and coastal with the best performance in pelagic. BST Chl-a improves coverage without significant quality degradation compared to the original MODIS Chl-a. BST Chl-a agrees better with in-situ data than that of MODIS, with CC of 0.742, RMSE of 0.247, MAE of 0.202 and Bias of 0.089. Cross-satellite validation using VIIRS and Himawari-8 Chl-a also shows promising results with the CC of 0.861 and 0.765, respectively, suggesting the high accuracy of BST Chl-a. The inter-annual trend of BST Chl-a decreases in coastal and increases in offshore and pelagic. BST Chl-a images present similar spatial patterns to MODIS Chl-a under different missing rates, with gradual decreases from coastal to pelagic. It indicates that phytoplankton bloom patterns can be identified by daily BST Chl-a images.

Polarized Micro-Raman Spectroscopy and 2D Convolutional Neural Network Applied to Structural Analysis and Discrimination of Breast Cancer.

Raman spectroscopy has been efficiently used to recognize breast cancer tissue by detecting the characteristic changes in tissue composition in cancerization. In addition to chemical composition, the change in bio-structure may be easily obtained via polarized micro-Raman spectroscopy, aiding in identifying the cancerization process and diagnosis. In this study, a polarized Raman spectral technique is employed to obtain rich structural features and, combined with deep learning technology, to achieve discrimination of breast cancer tissue. The results reconfirm that the orientation of collagen fibers changes from parallel to vertical during breast cancerization, and there are significant structural differences between cancerous and normal tissues, which is consistent with previous reports. Optical anisotropy of collagen fibers weakens in cancer tissue, which is closely related with the tumor's progression. To distinguish breast cancer tissue, a discrimination model is established based on a two-dimensional convolutional neural network (2D-CNN), where the input is a matrix containing the Raman spectra acquired at a set of linear polarization angles varying from 0° to 360°. As a result, an average discrimination accuracy of 96.01% for test samples is achieved, better than that of the KNN classifier and 1D-CNN that are based on non-polarized Raman spectra. This study implies that polarized Raman spectroscopy combined with 2D-CNN can effectively detect changes in the structure and components of tissues, innovatively improving the identification and automatic diagnosis of breast cancer with label-free probing and analysis.

Fibroblast growth factor (FGF), FGF receptor (FGFR), and cyclin D1 (CCND1) DNA methylation in head and neck squamous cell carcinomas is associated with transcriptional activity, gene amplification, human papillomavirus (HPV) status, and sensitivity to tyrosine kinase inhibitors.

BACKGROUND: Dysregulation of fibroblast growth factor receptor (FGFR) signaling pathway has been observed in head and neck squamous cell carcinoma (HNSCC) and is a promising therapeutic target for selective tyrosine kinase inhibitors (TKIs). Potential predictive biomarkers for response to FGFR-targeted therapies are urgently needed. Understanding the epigenetic regulation of FGF pathway related genes, i.e. FGFRs, FGFs, and CCND1, could enlighten the way towards biomarker-selected FGFR-targeted therapies. METHODS: We performed DNA methylation analysis of the encoding genes FGFR1, FGFR2, FGFR3, FGFR4, FGF1-14, FGF16-23, and CCND1 at single CpG site resolution (840 CpG sites) employing The Cancer Genome Research Atlas (TCGA) HNSCC cohort comprising N = 530 tumor tissue and N = 50 normal adjacent tissue samples. We correlated DNA methylation to mRNA expression with regard to human papilloma virus (HPV) and gene amplification status. Moreover, we investigated the correlation of methylation with sensitivity to the selective FGFR inhibitors PD 173074 and AZD4547 in N = 40 HPV(-) HNSCC cell lines. RESULTS: We found sequence-contextually nuanced CpG methylation patterns in concordance with epigenetically regulated genes. High methylation levels were predominantly found in the promoter flank and gene body region, while low methylation levels were present in the central promoter region for most of the analyzed CpG sites. FGFRs, FGFs, and CCND1 methylation differed significantly between tumor and normal adjacent tissue and was associated with HPV and gene amplification status. CCND1 promoter methylation correlated with CCND1 amplification. For most of the analyzed CpG sites, methylation levels correlated to mRNA expression in tumor tissue. Furthermore, we found significant correlations of DNA methylation of specific CpG sites with response to the FGFR1/3-selective inhibitors PD 173074 and AZD4547, predominantly within the transcription start site of CCND1. CONCLUSIONS: Our results suggest an epigenetic regulation of CCND1, FGFRs, and FGFs via DNA methylation in HNSCC and warrants further investigation of DNA methylation as a potential predictive biomarker for response to selective FGFR inhibitors in clinical trials.

On-chip monolithic Fourier transform spectrometers assisted by cGAN spectral prediction.

Silicon photonic spatial heterodyne Fourier transform spectrometers (SH-FTSs) are attractive with chip-scale monolithic arrays of imbalanced Mach-Zehnder interferometers; however, there exist optical path difference (OPD) errors from the inevitable fabrication imperfection, which will severely distort the retrieved spectra. In this Letter, we propose that a predictive model can be created for rapid and accurate spectral recovery based on the conditional generative adversarial network (cGAN) featuring strong input-on-output supervision, instead of both complicated physical OPD modification and time-consuming iterative spectral calculation. As a demonstration, cGAN spectral prediction was performed for our previously presented dual-polarized SH-FTS with large OPD errors [Opt. Lett.44, 2923 (2019)OPLEDP0146-959210.1364/OL.44.002923]. Due to the strong noise-resistant capability, the cGAN-predicted spectra can stay reliable, even though the signal-to-noise ratio of acquired interferograms dramatically drops from 1000 to 100, implying a lower limit of detection.

Classifying breast cancer tissue by Raman spectroscopy with one-dimensional convolutional neural network.

As the most common cancer in women, breast cancer is becoming lethal worldwide. However, the current breast diagnosis technologies are not enough to meet the requirements in clinic due to some shortages of early-stage insensitiveness, time consumption and relying on the doctor's experience, etc. It's necessary to develop a creative method for the automatical diagnosis of breast cancer. Therefore, Raman spectroscopy and one-dimensional convolutional neural network (1D-CNN) algorithm were combined together for the first time to classify the healthy and cancerous breast tissues in this study. First, a number of Raman spectra were collected from breast samples of 20 patients for spectral analysis. Then, a 1D-CNN model was developed and trained for classification. In addition, the Fisher Discrimination Analysis (FDA) and Support Vector Machine (SVM) classifiers were trained and tested with the same spectral data for comparison. The best classification performance, namely the overall diagnostic accuracy of 92%, the sensitivity of 98% and the specificity of 86%, has been achieved by using 1D-CNN model. This study proves that 1D-CNN combined with Raman spectroscopy can classify breast tissues effectively and automatically and lay the foundation for automated cancer diagnosis in the future.

FANCD2 knockdown with shRNA interference enhances the ionizing radiation sensitivity of nasopharyngeal carcinoma CNE-2 cells.

Fanconi anemia complementation group D2 (FANCD2) has been associated with the sensitivity of tumor cells to DNA crosslinking damaging agents in certain solid tumors. However, its role in nasopharyngeal carcinoma (NPC) is still unclear. In the present study, the role of FANCD2 in the response of NPC CNE-2 cells to radiation was investigated. A CNE-2 cell model with stable FANCD2 silencing was constructed by lentiviral transfection. Fluorescence quantitative PCR and western blotting were used to evaluate FANCD2 expression in CNE-2 cells. The biological impact of FANCD2 silencing on the response of CNE-2 cells to radiation was investigated in vitro and in vivo. The microarray technology, western blotting, and immunohistochemistry were used to analyze the proteins involved in related pathways after irradiation to investigate the underlying mechanism. Lentivirus-mediated shRNA interference stably silenced the FANCD2 gene in CNE-2 cells. In vitro, in the FANCD2-silenced group, cell proliferation was significantly inhibited, apoptosis was increased, and the cell cycle was arrested at the G2/M phase after irradiation. In vivo, FANCD2 silencing slowed tumor growth, as the volume and weight of the xenograft tumors were significantly decreased. Both in vitro and in vivo, the differentially expressed genes NUPR1, FLI1, and FGF21 were downregulated in the FANCD2-silenced group. Our results show that FANCD2 silencing affected the sensitivity of CNE-2 cells to ionizing radiation by regulating cell proliferation, apoptosis, and cell cycle distribution. The mechanism might be associated with changes in NUPR1, FLI1, and FGF21 protein expression due to the FANCD2 silencing. This study provides a promising target for NPC radiotherapy.

Effect of Blocking the OX40/OX40L Signaling Pathway by siRNA Interference on Animal Experimental Study of Allergic Rhinitis.

BACKGROUND: The identification of new approaches and intervention targets for the treatment of AR is urgently needed. We aimed to investigate the effect of blocking the OX40/OX40L signaling pathway by small interfering RNA (siRNA) on ovalbumin (OVA)-induced AR in a mouse model. METHODS: After establishment of the AR model, the mice were interfered by siRNA-OX40L (experimental group), siRNA-C (negative control group), or PBS (control group). Nose scratching, sneezing and nasal discharge were observed. OX40L mRNA and protein and the IL-5, TNF-α, regulatory T cell (Treg) -specific marker Foxp3, and eosinophil (EOS) levels were analyzed. RESULTS: The numbers of nose scratching and sneezing were significantly lower in the siRNA-OX40L-treated group (p <0.05). After the intervention of siRNA-OX40L, OX40L mRNA and protein levels were significantly inhibited (p <0.05), but the Foxp3 level was significantly increased in the experimental group (p <0.05). The IL-5 and TNF-α levels were significantly lower in the experimental group (p <0.05), and the reduction was more evident for the Th2-type cytokine IL-5 than for the Th1-type cytokine TNF-α. Few or no EOSs were found in the nasal mucosal epithelium of the experimental group (p <0.05), whereas EOS infiltration was significant in the other two groups. CONCLUSIONS: Blockage of the OX40/OX40L signaling pathway with siRNA-OX40L interference can inhibit allergic reactions and relieve allergic symptoms in AR mice. The underlying mechanism may be related to correcting Th2 immune deviation, inducing immune tolerance, and promoting Treg production.

Expression of FANCD2 is associated with prognosis in patients with nasopharyngeal carcinoma.

The relationship between Fanconi anemia complementation group D2 (FANCD2) and early diagnosis, pathogenesis, recurrence, and prognosis in patients with nasopharyngeal carcinoma (NPC) was investigated in a retrospective case-control study. The clinicopathological data of patients with NPC were collected. The results showed that FANCD2 was significantly higher in poorly differentiated squamous cell carcinoma than in moderately and well differentiated carcinoma. FANCD2 was significantly lower in recurrent NPC tissues than in NPC tissues before treatment. FANCD2 was markedly higher in T1-2, stage I-II NPC tissues with a duration of disease shorter than 6 months than in T3-4, stage III-IV NPC tissues with a duration of disease longer than 6 months. Moreover, compared with patients with cervical lymph node metastases, FANCD2 was elevated in tissues from patients without cervical lymph node metastases. Furthermore, the NPC patients in the high-FANCD2-expression group exhibited a higher recurrence rate than the patients in the low-FANCD2-expression group. Finally, the disease-free survival rate of the high-expression group was significantly lower than it was in the low-expression group. Therefore, FANCD2 is associated with the occurrence, differentiation, and cervical lymph node metastasis of NPC. With the development of NPC, FANCD2 is down-regulated. FANCD2 may be a molecular marker for the early diagnosis and prognosis of NPC.

Silencing of FANCD2 enhances the radiosensitivity of metastatic cervical lymph node-derived head and neck squamous cell carcinoma HSC-4 cells.

Fanconi anemia complementation group D2 (FANCD2) is involved in the key steps of the Fanconi anemia (FA) pathway, which plays a role in the repair of DNA crosslink damage. However, the role of FANCD2 during radiotherapy for head and neck squamous cell carcinoma (HNSCC) is unclear. In this study, the HNSCC cell line HSC-4 was used. Western blotting was used to evaluate the expression of the FANCD2 in HSC-4 cells. We investigated the impact of FANCD2 on the radiosensitivity of HSC-4 cells in vitro and in vivo. TUNEL, western blotting and immunohistochemistry were used to analyze the apoptosis and proteins involved in apoptosis-related pathways after radiotherapy to investigate the relevant mechanism. The present study showed that shRNA interference could effectively and stably silence FANCD2 expression in HSC-4 cells. In vitro, the silencing of FANCD2 inhibited cell proliferation, decreased the survival rate, increased apoptosis and induced S phase arrest in HSC-4 cells after radiotherapy. In vivo, the silencing of FANCD2 could prolong the tumor-forming time and slow tumor growth. In addition, the tumor volume was significantly reduced, the weight was deceased, and the tumor inhibition rate was increased after radiotherapy. TUNEL showed that the silencing of FANCD2 significantly increased apoptosis in HSC-4 cells induced by radiotherapy. Both in vitro and in vivo esperiments revealed that the expression of the Bax and p-p38 proteins in HSC-4 cells, in which FANCD2 had been silenced, was increased after radiotherapy, whereas the expression of the p38 and Bcl2 proteins was decreased. Our results suggested that the silencing of FANCD2 enhanced the radiosensitivity of HSC-4 cells, and its mechanism involves the activation of the p38 MAPK signaling pathway and the regulation of the expression of Bax and Bcl2 proteins. This study provides a novel candidate target for HNSCC therapy.