Search

Cancer-associated fibroblasts (CAFs) have been reported to play an essential role in tumor angiogenesis and progression. In this study, we aimed to investigate the impact of vocal fold leukoplakia-associated fibroblasts (VFLFs) on the angiogenesis process in vocal fold leukoplakia (VFL) and their potential secretions of proangiogenic factors.A total of 160 lesions (86 laryngeal carcinoma, 67 vocal fold leukoplakia, 7 vocal fold polyp) were detected under narrow band imaging (NBI) mode to evaluate the relationship between pathology and intraepithelial papillary capillary loop (IPCL) grades. We characterized immortalized vocal fold CAFs, VFLFs, normal fibroblasts (NFs) cell lines using immunofluorescence cytochemistry and real-time quantitative polymerase chain reaction (RT-qPCR). The effects of fibroblast conditioned media (CM) on the proliferative, migrating and tube formation capacity of human umbilical vein endothelial cells (HUVEC) were investigated using the cell counting kit-8 (CCK-8) assay, wound healing assay, transwell migration experiment and Matrigel tube formation experiment. The expression levels of proangiogenic factors in CAFs, VFLFs, and NFs were evaluated by antibody microarray and RT-qPCR.NBI images depicted that angiogenesis was abnormally activated during laryngeal tumorigenesis. Both CAF and VFLF expressed Vimentin, alpha-smooth muscle actin (α-SMA) and fibroblast activation protein (FAP). NF expressed Vimentin and α-SMA, but not FAP. The PCR results showed that mRNA expression levels of Vimentin, α-SMA and FAP in CAFs and VFLFs were significantly increased than those in NFs. CAF-CM and VFLF-CM promoted the proliferative, migrating, and tube formation ability of HUVECs. Secretome profiling of fibroblasts by antibody microarray demonstrated that VFLFs secreted significantly more vascular endothelial growth factor (VEGF), angiogenin, bFGF and HGF than NFs.Overall, we demonstrated that VEGF, angiogenin, bFGF and HGF derived from VFLFs may play crucial roles in the angiogenesis process of laryngeal premalignant and malignant lesions. This may contribute to the exploitation of novel therapeutic strategies for VFL.

More than 40% of lung cancers are lung adenocarcinoma (LUAD) worldwide. However, the prognosis of LUAD is poor for the lack of effective treatment methods. Our study identified PLK1 as a novel prognosis biomarker and treatment target for LUAD. Based on the Cancer Genome Atlas (TCGA) database, differentially expressed genes (DEGs) from 551 LUAD cases were analyzed for the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. To explore the biological pathways and the tumor-infiltrating immune cells (TICs) using gene set variation analysis (GSVA) and the CIBERSORT, as well as to analyze DEGs, a protein-protein interaction (PPI) network and Cox regression analysis were performed. Validation of DEGs was achieved through quantitative real-time PCR (qPCR) and immunoblotting. DEGs associated with the cell cycle were sorted out. Cell cycle scores were positively correlated with age, clinical stages, and metastasis and negatively correlated with overall survival of LUAD patients. PPI and Cox analyses showed that PLK1 could be a prognostic factor for LUAD patients. CIBERSORT analysis revealed a positive correlation between the transcription level of PLK1 and the function of CD8+ and activated memory CD4+ T cells, as well as a negative correlation with activated natural killer cells. Furthermore, PLK1 overexpression increased immune cytotoxicity, as measured by the cytolytic activity score, IFN- score, and IFN- level. There is a strong correlation between PLK1 and key features of TICs, indicating its potential as a promising prognostic biomarker for LUAD.

Since B7-H3 is overexpressed or amplified in many types of solid tumors with a restricted expression in the normal tissues, it has been an emerging immunotherapeutic target for solid tumors. This review will focus on the structural designs of developing chimeric antigen receptors (CARs) targeting B7-H3. The expression, receptor, and function of the B7-H3, as well as a short overview of B7-H3-targeted monoclonal antibody therapy, are discussed. Finally, a detailed summary of B7-H3 redirected CAR-T and CAR-NK cell approaches utilized in preclinical models and currently ongoing or completed clinical trials are presented. It has been demonstrated that B7-H3-targeted CAR-based cell therapies were safe in initial trials, but their efficacy was limited. Employing the local delivery routes, the introduction of novel modifications promoting CAR-T persistence, and combined treatment with other standard therapies could improve the efficacy of B7-H3-targeted CAR-T cell therapy against solid tumors.Cancers develop by avoiding being discovered and destructed by the immune system. Cancer immunotherapy is characterized as an approach to assist the immune system to recognize and fight cancer by using uses some substances. Chimeric antigen receptor (CAR) is a protein designed to recognize a target of interest on tumor cells, immune cells armed with which are able to treat human cancers. B7-H3 is a type I transmembrane protein overexpressed in various solid tumors but scarcely detected in normal tissues. Moreover, it is associated with a poor prognosis for many cancer patients. Therefore, we discussed the role of B7-H3 in solid tumors, and concluded it as a promising therapeutic target for CAR-based immunotherapy. A number of B7-H3 CARs have been developed and tested in recent years, but which CAR construct targeting B7-H3 works best remains uncertain. Up till now, the design of B7-H3 CARs is still controversial and challenging. Hence, we summarized and compared the designs and efficacies of B7-H3 CARs utilized in preclinical models and currently ongoing or completed clinical trials in this review, and aimed to provide a better reference for future B7-H3 CAR design.

PURPOSE: This study aimed to evaluate the value of long-read sequencing for preimplantation haplotype linkage analysis. METHODS: The genetic material of the three β-thalassemia mutation carrier couples was sequenced using single-molecule real-time sequencing in the 7.7-kb region of the HBB gene and a 7.4-kb region that partially overlapped with it to detect the presence of 17 common HBB gene mutations in the Chinese population and the haplotypes formed by the continuous array of single-nucleotide polymorphisms linked to these mutations. By using the same method to analyze multiple displacement amplification products of embryos from three families and comparing the results with those of the parents, it could be revealed whether the embryos carry disease-causing mutations without the need for a proband. RESULTS: The HBB gene mutations of the three couples were accurately detected, and the haplotype linked to the pathogenic site was successfully obtained without the need for a proband. A total of 68.75% (22/32) of embryos from the three families successfully underwent haplotype linkage analysis, and the results were consistent with the results of NGS-based mutation site detection. CONCLUSION: This study supports long-read sequencing as a potential tool for preimplantation haplotype linkage analysis. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10815-022-02415-1.

Ferroptosis is a novel form of cell death that is closely associated with the formation of many tumors. Our study focused on the mechanism by which long noncoding RNAs (lncRNAs) regulate ferroptosis in gastric cancer (GC) peritoneal metastasis (PM). We utilized lncRNA sequencing and protein profiling analysis to identify ferroptosis-associated lncRNAs and proteins. qRT-PCR was used to analyze the expression of BDNF-AS and FBXW7 in GC tissues and adjacent normal tissues. Chromatin isolation by RNA purification (ChIRP), RNA immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP), and coimmunoprecipitation (co-IP) assays were performed to investigate the interaction between BDNF-AS and its downstream targets. Finally, the function of BDNF-AS was validated

Mutations of the X-linked methyl-CpG-binding protein 2 (MECP2) gene in humans are responsible for most cases of Rett syndrome (RTT), an X-linked progressive neurological disorder. While genome-wide screens in clinical trials have revealed several putative RTT-associated mutations in MECP2, their causal relevance regarding the functional regulation of MeCP2 at the etiologic sites at the protein level requires more evidence. In this study, we demonstrated that MeCP2 was dynamically modified by O-linked-β-N-acetylglucosamine (O-GlcNAc) at threonine 203 (T203), an etiologic site in RTT patients. Disruption of the O-GlcNAcylation of MeCP2 specifically at T203 impaired dendrite development and spine maturation in cultured hippocampal neurons, and disrupted neuronal migration, dendritic spine morphogenesis, and caused dysfunction of synaptic transmission in the developing and juvenile mouse cerebral cortex. Mechanistically, genetic disruption of O-GlcNAcylation at T203 on MeCP2 decreased the neuronal activity-induced induction of Bdnf transcription. Our study highlights the critical role of MeCP2 T203 O-GlcNAcylation in neural development and synaptic transmission potentially via brain-derived neurotrophic factor. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12264-021-00784-8.

Tumor-infiltrating immune cells have been implicated in the tumorigenesis and progression of esophageal squamous cell carcinoma (ESCC). However, the functionalities and clinical significance of immune cells remain largely unveiled. In this study, the gene expression data from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were extracted. The relative infiltrating levels were estimated by single-sample gene set enrichment analysis. Some cytotoxic immune cells were attenuated, and resting cytotoxic immune cells were accumulated in ESCC. Remarkably, we also observed that infiltrating levels of macrophage M2 and resting natural killer (NK) cells were increased in nonresponders of CRT, and T cells that had anticancer activities such as activated memory CD4 and T helper 2 (Th2) cells were significantly reduced in ESCC tissues of the nonresponders. Moreover, the high infiltrations of the resting natural killer (NK) and dendritic cell (DC) were observed to result in a shorter overall survival in ESCC. Consistently, high expression of immune checkpoint genes, CTLA4 and HAVCR2, was associated with poor prognosis. Furthermore, STAT5B, a key transcription factor, as well as its target genes, involved in the regulation of T cells, was significantly downregulated in ESCC, especially subgroup I, indicating that downregulation of STAT5B might be associated with reduced T cell-mediated anticancer activity. In conclusion, the present study significantly improved our understanding of the regulatory roles of immune cells in ESCC.

Indium, a low melting point metal, is well-known for constructing eutectic gallium-indium liquid metal. However, unlike liquid metal nanoparticles, the biomedical applications of metallic indium nanoparticles (In NPs) remain in their infancy. Herein, an ultrasound-assisted liquid-reduction synthesis strategy was developed to prepare PEGylated In NPs, which were then used as a high-performance contrast agent for enhancing multiwavelength photoacoustic imaging and second near-infrared (NIR-II) photothermal therapy of the 4T1 breast tumor. The obtained In NPs depicted remarkable optical absorption from the first near-infrared (NIR-I) to NIR-II region and a high photothermal conversion efficiency of 41.3% at 1064 nm, higher than the majority of conventional NIR-II photothermal agents. Upon injection into the tumor, the photoacoustic intensities of the tumor section post-injection were obviously increased by 2.59-, 2.62-, and 4.27-fold of those of pre-injection by using excitation wavelengths of 750, 808, and 970 nm, respectively, depicting an excellent multiwavelength contrast capability of photoacoustic imaging. In addition, efficient ablation of the 4T1 tumor was achieved through the photothermal performance of PEGylated In NPs under NIR-II laser irradiation. Importantly, as the widely used element in the clinic, In NPs were highly biocompatible in vitro and in vivo. Therefore, this work pioneered the biomedical applications of PEGylated In NPs for cancer diagnosis and treatment.

Introduction Medulloblastoma (MB) is a malignant pediatric brain tumor that arises in the cerebellar granular neurons. Sonic Hedgehog subtype of MB (SHH‐MB) is one of the major subtypes of MB in the clinic. However, the molecular mechanisms underlying MB tumorigenesis are still not fully understood. Aims Our previous work demonstrated that the receptor for activated C kinase 1 (Rack1) is essential for SHH signaling activation in granule neuron progenitors (GNPs) during cerebellar development. To investigate the potential role of Rack1 in MB development, human MB tissue array and SHH‐MB genetic mouse model were used to study the expression of function of Rack1 in MB pathogenesis. Results We found that the expression of Rack1 was significantly upregulated in the majority of human cerebellar MB tumors. Genetic ablation of Rack1 expression in SHH‐MB tumor mice could significantly inhibit MB proliferation, reduce the tumor size, and prolong the survival of tumor rescue mice. Interestingly, neither apoptosis nor autophagy levels were affected in Rack1‐deletion rescue mice compared to WT mice, but the expression of Gli1 and HDAC2 was significantly decreased suggesting the inactivation of SHH signaling pathway in rescue mice. Conclusion Our results demonstrated that Rack1 may serve as a potential candidate for the diagnostic marker and therapeutic target of MB, including SHH‐MB., Rack1 is essential for GNPs development by activation of Shh signaling. Here, we found that Rack1 was significantly upregulated in some cerebellar MB tumor samples. Genetic ablation of Rack1 in SHH‐subtype medulloblastoma (MB) tumor mice could significantly reduce tumor cell proliferation, reduce the tumor size, and prolong the survival of tumor rescue mice, indicating that Rack1 may serve as a potential novel therapeutic target for SHH‐type MB in the clinic.

Studies of saccular and utricular function in patients with obstructive sleep apnea (OSA) are rare. We noticed that some OSA patients also had positive results in vestibular function tests, and this inspired our interest in exploring the vestibular function patterns of OSA patients.To investigate otolithic organ function in severe OSA patients who lack vestibular symptoms and systemic disease.32 patients (64 ears) with severe OSA and 22 healthy controls (44 ears) were enrolled. The ocular and cervical vestibular-evoked myogenic potentials (oVEMP and cVEMP) and the caloric test were recorded and analyzed.The response rates of oVEMP (73.4%) and cVEMP (82.8%) in patients with severe OSA were significantly lower than those in controls. In oVEMP, elevated thresholds (Disappearance or impairment of VEMPs could be observed in patients with severe OSA, and reflects different degrees of impairment in the utricle and saccule.

The outbreak of novel coronavirus (COVID-19) has become a global concern that is deteriorating environmental quality and damaging human health. Though some researchers have investigated the linkage between temperature and COVID-19 transmissibility across different geographical locations and over time, yet these studies are scarce. This study aims to bridge this gap using daily temperature and COVID-19 cases (transmissibility) by employing grey incidence analysis (GIA) models (i.e., Deng’s grey incidence analysis (DGIA), the absolute degree GIA (ADGIA), the second synthetic degree GIA (SSDGIA), the conservative (maximin) model) and correlation analysis. Data on temperature are accessed from the NASA database, while the data on COVID-19 cases are collected from the official website of the government of Pakistan. Empirical results reveal the existence of linkages between temperature and COVID-19 in all Pakistani provinces. These linkages vary from a relatively stronger to a relatively weaker linkage. Based on calculated weights, the strength of linkages is ranked across provinces as follows: Gilgit Baltistan (0.715301) > Baluchistan (0.675091) > Khyber Pakhtunkhwa (0.619893) > Punjab (0.619286) > Sindh (0.601736). The disparity in the strength of linkage among provinces is explained by the discrepancy in the intensity of temperature. Besides, the diagrammatic correlation analysis shows that temperature is inversely linked to COVID-19 cases (per million persons) over time, implying that low temperatures are associated with high COVID-19 transmissibility and vice versa. This study is among the first of its kind to consider the linkages between temperature and COVID-19 transmissibility for a tropical climate country (Pakistan) using the advanced GIA models. Research findings provide an up-to-date glimpse of the outbreak and emphasize the need to raise public awareness about the devastating impacts of the COVID-19. The educational syllabus should provide information on the causes, signs, and precautions of the pandemic. Additionally, individuals should practice handwashing, social distancing, personal hygiene, mask-wearing, and the use of hand sanitizers to ensure a secure and supportive atmosphere for preventing and controlling the current pandemic.

The aim of this study was to evaluate the influence of preoperative tracheotomy on oncologic outcomes of advanced stage glottic carcinoma patients, and to explore the potential reason. We retrospectively analyzed 413 consecutive advanced stage glottic carcinoma patients from January 2005 to December 2010. The correlation of preoperative tracheotomy and potential impacting factor of tumor size involving tumor diameter and tumor area with overall survival (OS) and disease-free survival (DFS) was fully assessed. Our cohort consisted of 302 (73.1%) patients with T3 and 111 (26.9%) patients with T4, and 98 (23.7%) patients received preoperative tracheotomy. The OS and DFS rates of patients receiving preoperative tracheotomy were worse than those without (5-year OS: 49.3% versus 69.8%; 5-year DFS: 45.3% versus 61.0%). The mean tumor diameter and tumor area of patients with preoperative tracheotomy were greater than those without (3.3 cm versus 2.4 cm, 8.9 cm2 versus 4.7 cm2). The optimal cutoff values of tumor diameter and tumor area for tracheotomy were 2.85 cm and 6.64 cm2. Tumor diameter and tumor area were correlated with tracheotomy intervention. Furthermore, when considering the potential effect of tumor area in multivariate model, we found that it was a significant factor in survival outcomes but variable of preoperative tracheotomy was not. This study indicates that tumor size is correlated with preoperative tracheotomy, and tracheotomy intervention may be reflection from effect of great tumor size that is a true adverse factor influencing oncologic outcomes of advanced stage glottic carcinoma patients.

It is important for the patients to reconstruct the voice phonic function by surgery after total laryngectomy in the developing countries.To investigate the clinical outcomes of voice reconstruction using an infrahyoid musculocutaneous flap for patients after total laryngectomies.Eighteen male patients recruited were laryngectomized. The infrahyoid musculocutaneous flap was designed. After total laryngectomy, the lower edge of the flap was sewed with the upper edge of the tracheostomy opening. Next, the lateral and medial edges of the flap were anastomosed to create a pronunciation tube. Finally, the remaining opening of the tube was sutured with the anterolateral wall of the hypopharynx to establish a communication with the pharyngeal cavity.A total of 17 cases of flaps were survived and only 1 necrosed. There were 17 patients without serious complications, except that 6 cases had mild irritable cough when gulping water. However, it could be relieved through blocking tracheostoma. One year after operation, all patients could more remarkably articulate clear, powerful, and consistent words. The articulatory configuration was existed under rigid laryngoscope and CT.The use of an infrahyoid myocutaneous flap is feasible for the voice restoration in the patients undergoing total laryngectomy.

The purpose of this study was to evaluate the potential predictor of tumor size on rates of overall and disease-free survival (OS and DFS) as determined by postoperative pathologic examination in patients with glottic carcinoma.Retrospective cohort study.Tertiary care university hospital.In this study, 1337 consecutive patients with glottic carcinoma who underwent surgical treatment from 2005 to 2010 were retrospectively reviewed. The influence of tumor size that was evaluated by tumor area (tumor length × tumor width) on OS and DFS outcomes was assessed by Cox regression analyses.In all, 1303 (97.5%) patients were male, and 34 (2.5%) were female, with a mean ± SD age of 60.4 ± 10 years. The 10-year OS and DFS rates were 72.9% and 69.9%, respectively. The tumor area cutoff values that best discriminated OS and DFS rates were both 1.80 cmThe results of this study indicate that patients with glottic carcinoma with a tumor area1.8 cm

Objectives Previous evidence suggested Helicobacter pylori (H. pylori) promoted tumorigenesis by inducing inflammation in gastric premalignant disease. Whether this organism affected the development of head and neck precancerous lesion remains unknown. We performed this study to assess the prognostic impact of H. pylori infection on patients with laryngeal precancerous lesion. Methods In this prospective study, 41 enrolled patients with laryngeal precancerous lesion were arranged into H. pylori-positive group (n = 16) and H. pylori-negative group (n = 25). Two poor outcomes were recurrence and malignant transformation. Prognostic impact of H. pylori infection was analyzed using the Univariate Cox proportional hazard model and Cochran–Mantel–Haenszel test. We also analyzed the prognostic value of systemic inflammation-based markers. Results During a median follow-up of 32.9 months, recurrence developed in 8 patients in the H. pylori positive group and 5 patients in the H. pylori negative group (50.0% vs. 20.0%, P = 0.026, Log-rank test). Malignant transformation developed in 4 patients in the H. pylori positive group and no patients in the H. pylori negative group (25.0% vs. 0.0%, P = 0.005, Log-rank test). H. pylori positive group had a significant higher malignant transformation rate after stratifying on the observed covariates. There was no significant difference of recurrence rate in two groups stratified with laryngopharyngeal reflux, size of lesion, and pathological type (Cochran–Mantel–Haenszel test). The basal levels of lymphocyte counts and neutrophils to lymphocytes ratio were significantly different in patients with H. pylori infection. Moreover, the receiver operating characteristic curve analysis revealed a predictive value of lymphocyte counts and neutrophils to lymphocytes ratio for poor outcomes (areas under the curve, 0.658 and 0.651, respectively). Conclusion The data demonstrate a malignant transformation-relevant role of H. pylori in laryngeal precancerous lesion. H. pylori infection combined with laryngopharyngeal reflux, size of lesion, and pathological type might relate to recurrence of laryngeal precancerous lesion. And changes of lymphocyte counts and neutrophils to lymphocytes ratio associated with H. pylori infection might deserve as predictive indicators for poor prognosis of laryngeal precancerous lesion.

The characteristics of fibroblast cells in head and neck precancerous lesion and its ability to secrete inflammatory cytokines and affect CD8

The molecular mechanism underlying gastric cancer (GC) peritoneal metastasis (PM) remains unclear. Here, we identified LINC00924 as a GC PM-related lncRNA through Microarray sequencing. LINC00924 was highly expressed in GC, and its high expression is associated with a broad range of PM. Via RNA sequencing, RNA pulldown assay, mass spectrometry, Seahorse, Lipidomics, spheroid formation and cell viability assays, we found that LINC00924 promoted fatty acid (FA) oxidation (FAO) and FA uptake, which was essential for matrix-detached GC cell survival and spheroid formation. Regarding the mechanism, LINC00924 regulated the alternative splicing (AS) of Mnk2 pre-mRNA by binding to hnRNPC. Specifically, LINC00924 enhanced the binding of hnRNPC to Mnk2 pre-mRNA at e14a, thus downregulating Mnk2a splicing and regulating the p38 MAPK/PPARα signaling pathway. Collectively, our results demonstrate that LINC00924 plays a role in promoting GC PM and could serve as a drug target.

Photothermal-enhanced chemodynamic therapy (CDT) has been attracting increasing attention for effective tumour treatment. Nevertheless, even though Mn-based nanostructures are promising CDT agents, their photothermal conversion capacities are not good enough for an ideal combination therapy. In this work, a bifunctional Bi

Circulating tumor cells (CTCs) are important precursors of colorectal cancer (CRC) metastasis. The epithelial-mesenchymal transition (EMT) process facilitates CTC invasion by allowing these cells to evade antimetastatic checkpoints to mediate distant metastasis. However, the specific molecular mechanism of tumor EMT remains largely unknown. Based on our previous research on the YAP1 pathway, we further studied the upstream molecule small nucleolar RNA host gene 16 (SNHG16), whose expression was correlated with advanced TNM stage, distant metastasis, and poor prognosis in CRC patients. Furthermore, loss- and gain-of-function assays revealed that SNHG16 promoted CRC colony formation, proliferation, migration, invasion, EMT, mesenchymal-like CTC generation, and liver metastasis through YAP1. Mechanistically, SNHG16 acted as a miRNA sponge to sequester miR-195-5p on Ago2, thereby protecting YAP1 from repression. Moreover, YAP1 bound TEA domain transcription factor 1 (TEAD1) to form a YAP1/TEAD1 complex, which in turn bound two sites in the promoter of SNHG16 and regulate SNHG16 transcription. Finally

OBJECTIVE: This study investigated the clinical indications, prevention and management of common complications of TORS resection of parapharyngeal space tumors. METHODS: The clinical data of 23 patients with parapharyngeal space tumors treated with TORS in the Eye & ENT Hospital of Fudan University from July 2020 to February 2022 were retrospectively analyzed. The surgical methods were divided into simple TORS surgery and TORS combined endoscopic or cervical surgery. The surgical incision can be directly sutured or partially sutured. If the surgical cavity is below the oropharyngeal plane and there is a risk of airway compression, preventive tracheotomy would be performed. No drainage(15 cases) or transnasal negative pressure drainage tube(8 cases) was placed in the operation cavity. In 17 cases, tumors were located between the top of nasopharynx and the lower boundary of oropharynx and these patients underwent simple TORS surgery; one case received combined cervical and endoscopic surgery, as the tumor was located near the skull base with unclear boundary with the deep lobe of parotid gland; five cases underwent combined endoscopic surgery, as the tumor reached the upper part of the nasopharynx or the outer part of the parapharyngeal space. RESULTS: One case underwent emergency hemostasis for postoperative bleeding, and three cases underwent puncture and drainage for postoperative effusion. No complication occurred such as postoperative infection and airway obstruction. The healing grade of surgical incision and surgical cavity in transnasal drainage group was significantly better than that in non-drainage group. CONCLUSION: TORS operation is a safe and minimally invasive method in treating parapharyngeal space tumors with appropriate size and location. Postoperative effusion is a common complication after TORS. In case that surgical cavity extends to the deep parapharyngeal space or the lower part of the surgical cavity is beneath the surgical incision, the placement of transnasal negative pressure drainage tube after operation can improve postoperative recovery and reduce the incidence of complications such as effusion and infection.

At present, the dual pressure of rural labor outflow and population aging in China makes the problems of the rural elderly population increasingly prominent, and its health problem is particularly prominent. Based on the 2014 China elderly population health survey data (CLHLS), this paper finds that the physical health status of the rural elderly has a significant positive impact on their loneliness; that is, the rural elderly with poor health status are more likely to feel lonely. At the same time, the age of the elderly has a significant positive impact on their loneliness. On the contrary, gender, personality, family income and intergenerational support of the elderly have a negative impact on their loneliness. Chronic diseases such as hypertension and diabetes have no significant effect on the loneliness of the elderly in rural areas, but there is a “severe disease effect”; that is, when chronic diseases develop into serious diseases or acute serious diseases, it can negatively impact the elderly psychologically and produce or deepen their sense of loneliness. Based on the above conclusions, this paper further puts forward relevant policy suggestions from three aspects: constructing a disease prevention and control system for the rural elderly, improving the care and service system for the rural elderly, reshaping rural filial piety culture, and creating a good atmosphere of “respecting, loving and respecting parents” in rural areas.

The study develops a theoretical framework of how irrigation and drainage infrastructure and rural transportation infrastructure influence poverty. Using panel data on 31 provinces in China from 2002 to 2017, this paper estimates basic and continuous difference-in-differences (DID) models to investigate the preliminary impact of irrigation and drainage infrastructure and rural transportation infrastructure on poverty and further explores the influence mechanisms of these rural infrastructures on poverty by using the mediating effect model. The results show that irrigation and drainage facilities infrastructure can directly reduce poverty. On the one hand, rural transportation infrastructure directly leads to rural hollowing out and aggravates rural poverty; on the other hand, it indirectly promotes poverty reduction by stimulating economic growth. Overall, the positive and negative effects of rural transportation infrastructure on poverty offset each other.

The pathology of vocal fold leukoplakia (VFL) was hard to predict before surgery.To investigate the correlation of intraepithelial papillary capillary loops (IPCLs) with the malignant transformation in VFL.The IPCLs in 90 patients were classified according to 2019 Ni classification. Type I-III represented to non- to moderate dysplasia in pathology being viewed as 'benign leukoplasia', type IV-VI represented to severe dysplasia/cancerForty-one cases were classified as type I-III, the consistency of type I-III with pathology was 61.0%, with 100.0% of type I, 47.1% of type II and 30.0% of type III. 49 cases were classified as type IV-VI, all were 'malignant leukoplasia', the consistency of type IV-VI and pathology was 100.0%. The accuracy of type I-III, IV-VI to predict pathology were 61.0, 100.0%. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value of Ni classification corresponding to 'benign/malignant leukoplasia' was 75.8, 100.0, 82.2, 100.0, 60.0%, respectively.IPCLs of type IV-VI in vocal cord leukoplasia had a close correlation with pathology. The degree of dysplasia was underestimated in type II and type III.

Despite attracting increasing attention in clinic, non-invasive high-intensity focused ultrasound (HIFU) surgery still commonly suffers from tumor recurrence and even matastasis due to the generation of thermo-resistance in non-apoptotic tumor cells and adverse therapy-induced inflammation with enhanced secretion of growth factors in irradiated region. In this work, inspired by the intrinsic property that the expression of thermo-resistant heat shock proteins (HSPs) is highly dependent with adenosine triphosphate (ATP), dual-functionalized diclofenac (DC) with anti-inflammation and glycolysis-inhibition abilities was successfully co-encapsulated with phase-change dl-menthol (DLM) in poly(lactic-co-glycolic acid) nanoparticles (DC/DLM@PLGA NPs) to realize improved HIFU surgery without causing adverse inflammation. Both in vitro and in vivo studies demonstrated the great potential of DC/DLM@PLGA NPs for serving as an efficient synergistic agent for HIFU surgery, which can not only amplify HIFU ablation efficacy through DLM vaporization-induced energy deposition but also simultaneously sensitize tumor cells to hyperthermia by glycolysis inhibition as well as diminished inflammation. Thus, our study provides an efficient strategy for simultaneously improving the curative efficiency and diminishing the harmful inflammatory responses of clinical HIFU surgery. Graphical Abstract

Peritoneal metastasis (PM) is one of the main causes of a poor prognosis in patients with advanced gastric cancer (GC). lncRNAs have been confirmed to play a very crucial role in the occurrence, development, and metastasis of many human cancers, including gastric cancer. However, the mechanism of lncRNA in PM of GC is rarely studied. We explored the mechanism of PM of GC through lncRNA gene sequencing and protein profiling analysis to detect PM-associated lncRNAs and proteins. A quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to identify the mRNA expression of SEMA3B-AS1 and BGN in GC tissues and adjacent normal tissues. The biological function of SEMA3B-AS1 in the PM of GC was identified through gain- and loss-of-function assays. Chromatin isolation by RNA purification (ChIRP), RNA immunoprecipitation (RIP), RNA pull-down, luciferase reporter, and coimmunoprecipitation (co-IP) assays was carried out to demonstrate the potential mechanism between SEMA3B-AS1 and its downstream genes, including HMGB1, FBXW7, and BGN. Finally, the biological function of SEMA3B-AS1 was demonstrated in animal experiments. The mRNA expression level of SEMA3B-AS1 was downregulated in GC and PM tissues compared to normal stomach tissues; however, BGN was highly expressed at the mRNA level. SEMA3B-AS1 was closely related to PM and the overall survival (OS) of GC patients. Functionally, the overexpression of SEMA3B-AS1 was related to GC progression, PM, and prognosis. Mechanistically, SEMA3B-AS1 could combine with HMGB1 to regulate the transcription of FBXW7, thus facilitating the ubiquitination of BGN. In conclusion, our study demonstrated that the SEMA3B-AS1/HMGB1/FBXW7 axis plays an inhibitory role in the PM of GC by regulating BGN protein ubiquitination. It also provides a new biological marker for the diagnosis and treatment of the PM of GC.

Endoscopic keel placement is difficult for patients with poor laryngeal exposures.To introduce a novel extra-endo laryngeal puncture technique for endoscopic keel placement to manage anterior glottic webs.Patients with anterior glottic webs and difficult laryngeal exposure in our hospital during 2018 to 2020 were recruited in this study. After the incision of the web, a novel puncture needle modified from a maxillary sinus puncture needle was inserted into endolarynx, and could pull the suture passing along a keel from inside out. The suture was then fixated on the neck, and the keel was placed for 4 weeks. Patients were followed up for at least 1 year.Eight patients including three men and five women were included in this study. All the patients tolerated the silicone keel well, and 7 of 8 (87.5%) patients did not have web relapses and reached remarkable voice improvements during follow-up. None of the cases needed a tracheotomy due to the dyspnea postoperatively.The novel puncture technique is an easy and effective approach to facilitate endoscopic keel placement for the treatment of anterior glottic webs and other surgical procedures requiring endolaryngeal suturing.

Introduction: There is still no general method for discriminating between benign and malignant leukoplakia and identifying vocal fold leukoplakia. Objective: To evaluate the reliability of a morphological classification and the correlation between morphological types and pathological grades of vocal fold leukoplakia. Methods: A total of 375 patients with vocal fold leukoplakia between 2009 and 2015 were retrospectively reviewed. Two observers divided the vocal fold leukoplakia into flat and smooth, elevated and smooth, and rough type on the basis of morphological appearance. The inter-observer reliability was evaluated and the results of classification from both observers were compared with final pathological grades. Clinical characteristics between low risk and high risk group were also analyzed. Results: The percentage inter-observer agreement of the morphological classification was 78.7% (κ = 0.615, p

To evaluate the predictive value of preoperative peripheral inflammatory markers in patients with vocal fold leukoplakia. A retrospective study was performed of the patients diagnosed with vocal fold leukoplakia and who accepted carbon dioxide (CO2) laser resection in our center in the last 10 years. We calculated the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and monocyte-to-lymphocyte ratio (MLR) after collecting and analyzing the clinical, histopathological and laboratory data. The potential relation between blood indexes and clinical events as recurrence or canceration was evaluated. A total of 589 patients were involved, including 300 cases without recurrence (group A), 198 with recurrence but not canceration (group B) and 91 transformed into squamous cancer (group C). Baseline analysis of NLR, PLR, and MLR showed no difference among the three groups before the first surgery. But all the indexes significantly elevated in groups B (P

Background: Laryngeal carcinoma should be treated with the intent of organ-sparing, and supracricoid partial laryngectomy with cricohyoidoepiglottopexy (CHEP) might be an important option.A...

Background: The effect of surgical margins on local recurrence in T1 glottic squamous cell carcinoma (GSCC) is unclear.Objectives: To investigate the association between surgical margins and local ...

To investigate the anatomy of the infratemporal fossa (ITF) and to discuss the practicality of endoscopic transvestibular surgery for an ITF tumor. Five fresh cadaveric specimens (10 sides) with vascular silicone injection were prepared for endoscopic anatomy. A transvestibular vertical incision was made along the ramus of the mandible, and pivotal nerves, arteries, and muscles were exposed to sculpt the anatomic landmarks of the ITF. The endoscopic transvestibular approach exposed the detailed structure of the ITF. The buccinator muscle and the adjoining superior pharyngeal constrictor muscle shaped the paramedian border of the ITF, while the medial pterygoid muscle (MPM) and the lateral pterygoid muscle formed the lateral border. The ITF was delimited by the skull base in the upper margin, and it was proximal to the parapharyngeal space in the inferior part. The inferior alveolar nerve was the first reference point, and the maxillary artery and the lateral pterygoid muscle were also the landmarks of the ITF. The lingual nerve, the eustachian tube (ET), and the middle meningeal artery were also located in the posterior part of the ITF. The endoscopic transvestibular approach provides a feasible and facile corridor to the ITF. With accurate hemostasis, this approach may provide another option for accessing the ITF for removal of tumors.

Migrants workers are important participants in and contributors to economic and social construction, but they still face the reality of being marginalized. Based on data from the China Migrants Dynamic Survey in 2018, this paper systematically investigated the impact of public health services on the multidimensional poverty of migrant workers. The research found that, first, the current mean of the multidimensional poverty deprivation value of migrant workers is 0.1806, which is one dimension of poverty that exists on average. In addition, migrant workers do not have high access to public health services. The proportions of migrant workers who have not established residents’ health files and who have not received public health education are 74.22% and 29.92%, respectively. Second, public health services can significantly alleviate the multidimensional poverty of migrant workers. After mitigating the potential endogeneity problem by the IV-2SLS method and conducting robustness tests by the PSM method, the conclusion is still robust. Further research found that the impact of public health services on the multidimensional poverty alleviation of migrant workers is heterogeneous. The improvement of public health services has the greatest effect on the multidimensional poverty alleviation of the new generation of migrant female workers in the western region. The research in this paper helps to examine and clarify the policy significance of public health services for the multidimensional poverty alleviation of migrant workers and provides empirical evidence for the use of public health services to tackle the poverty problem.

Environmental emergencies are associated with great harm, sudden onset and high publicity and cause serious damage to a broad range of ecological environmental, human health and social properties in a short period of time. Environmental decentralization can help local governments strengthen their independence and make better use of the advantages of environmental information to curb the occurrence of environmental emergencies, but regional corruption significantly weakens its effectiveness. Therefore, based on panel data of China's 30 provincial administrative regions between 2005 and 2016, this paper applies the panel threshold model to explore the relationship between environmental emergencies, environmental decentralization, and regional corruption. The results indicate that, first, environmental decentralization, environmental administrative decentralization, environmental monitoring decentralization, and environmental supervision decentralization all have a negative influence on environmental emergencies; second, as the degree of corruption in a region increases, the effect of environmental decentralization on restraining environmental emergencies decreases; and finally, there is heterogeneity in the relationship between environmental emergencies and environmental decentralization in various regions. Environmental decentralization in the eastern and western regions negatively affects environmental emergencies, while there is a positive relationship between the two in the central region.

Background Approximately 5.0–24.2% of colorectal cancers (CRCs) have inactivating mutations in SMAD4, making it one of the frequently mutated genes in CRC. We thus carried out a comprehensive system review and meta-analysis investigating the prognostic significance and clinicopathological features of SMAD4 gene mutation in CRC patients. Methods A detailed literature search was conducted in PubMed, Web of Science and Embase databases to study the relationship between SMAD4 mutations and the demographic and clinicopathological characteristics in CRC patients. The hazard ratios (HRs) with 95% confidence intervals (CI) were used to evaluate the effect of SMAD4 mutations on overall survival (OS) and progression-free survival (PFS)/recurrence-free survival (RFS). Results Ten studies enrolling 4394 patients were eligible for inclusion. Data on OS were available from 5 studies and data on PFS/RFS were available from 3 studies. Comparing SMAD4-mutated CRC patients with SMAD4 wild-type CRC patients, the summary HR for OS was 1.46 (95% CI 1.28–1.67, P = 0.001), the summary HR for PFS/RFS was 1.59 (95% CI 1.14–2.22, P = 0.006). In terms of clinicopathology parameters, 9 studies have data that can be extracted, SMAD4 mutations were associated with tumor location (odds ratio [OR] = 1.15, colon/rectum, 95% CI 1.01–1.31, P = 0.042), TNM stage (OR = 1.28, stage IV/I–III, 95% CI 1.03–1.58, P = 0.025), lymph node metastasis (OR = 1.42, N1 + N2/N0, 95% CI 1.20–1.67, P P P = 0.001). No connection was found with age, gender, tumor grade, microsatellite instability status and b-viral oncogene homolog B1 mutation status. Besides, publication bias was not observed in any study. Conclusions This meta-analysis suggests that SMAD4 mutation was associated with OS, PFS/RFS, and clinicopathological parameters, including tumor site, disease stage, RAS status, lymph node metastasis and mucinous differentiation. Our meta-analysis indicated that SMAD4 mutations could predict the poor prognosis and aggressive clinicopathological characteristics of CRC. More large-sample cohort studies are needed to confirm this conclusion. Since SMAD4 mutations are closely related to RAS mutations, their relationship warrants further investigation.

Since human beings have a long tradition of coexistence with pandemics, which may profoundly impact them, adopting preventive measures is crucial for humankind’s survival. This study explores the intention-based critical factors affecting the willingness of individuals to adopt pandemic prevention. To this end, a representative sample of 931 Pakistanis filled in an online questionnaire. However, only 828 questionnaires were found to be complete and valid for path modeling analysis. The core findings are as follows: Firstly, peer groups’ beliefs, self-efficacy, perceived risk, pandemic knowledge, ease of pandemic prevention adoption, and risk-averse behavior are revealed as driving forces of the individuals’ willingness to adopt pandemic prevention. Contrastingly, a lack of trust in political will and mythical attitude towards pandemics are uncovered as inhibitors. Nevertheless, moral values depict a neutral role. Secondly, the peer groups’ beliefs are highest ranked, followed by the lack of trust in political will and a mythical attitude towards pandemic prevention. Finally, moral values are determined as the lowest-ranked critical factor. Based on these results, the government should promote awareness campaigns on lethality and fatality of the pandemic at both centralized and decentralized levels to win people’s trust at the grass-roots level and overcome the mythical attitude of individuals at all societal levels. Besides, access to personal protective gears should be made feasible since an easier pandemic prevention adoption would increase the individuals’ willingness to adopt such preventative measures.

Aversive memories are long-lasting and prone to have adverse effects on our emotional wellbeing and mental health. Yet, how to remedy the maladaptive effects of aversive memories remains elusive. Using memory reactivation and emotional updating manipulations, we investigated how positive and neutral emotion updates aversive memories for reconsolidation in humans. We found that positive updating after reactivation was equivalent to neutral updating in altering true memories of the aversive story, but introduced more false memory. Moreover, an additional 12 hours of sleep reconsolidation did not further enlarge true memory differences, but attenuated the effect of reactivation and updating on false memory. Interestingly, the neutral rather than the positive updating reduced the emotional arousal of the aversive memory 24 hours later. Our findings provide novel insights into real-world therapeutic applications regarding how updating with positive and neutral emotion may reshape aversive memories, especially when taking wake- and sleep-filled reconsolidation into account.

Background Human bone marrow-derived mesenchymal stem cells (hBMSCs) have chondrocyte differentiation potential and are considered to be a cell source for cell-transplantation-mediated repair of cartilage defects, including those associated with osteoarthritis (OA). However, chondrocyte hypertrophic differentiation is a major obstacle for the application of hBMSCs in articular cartilage defect treatment. We have previously shown that microRNA-27b (miR-27b) inhibits hypertrophy of chondrocytes from rat knee cartilage. In this study, we investigated the role of miR-27b in chondrocyte hypertrophic differentiation of hBMSCs. Methods Chondrogenic marker and microRNA expression in hBMSC chondrogenic pellets were evaluated using RT-qPCR and immunohistochemistry. The hBMSCs were transfected with miR-27b before inducing differentiation. Gene and protein expression levels were analyzed using RT-qPCR and western blot. Coimmunoprecipitation was used to confirm interaction between CBFB and RUNX2. Luciferase reporter assays were used to demonstrate that CBFB is a miR-27b target. Chondrogenic differentiation was evaluated in hBMSCs treated with shRNA targeting CBFB. Chondrogenic hBMSC pellets overexpressing miR-27b were implanted into cartilage lesions in model rats; therapeutic effects were assessed based on histology and immunohistochemistry. Results The hBMSCs showed typical MSC differentiation potentials. During chondrogenic differentiation, collagen 2 and 10 (COL2 and COL10), SOX9, and RUNX2 expression was upregulated. Expression of miR-140, miR-143, and miR-181a increased over time, whereas miR-27b and miR-221 were downregulated. Cartilage derived from hBMSC and overexpressing miR-27b exhibited higher expression of COL2 and SOX9, but lower expression of COL10, RUNX2, and CBFB than did the control cartilage. CBFB and RUNX2 formed a complex, and CBFB was identified as a novel miR-27b target. CBFB knockdown by shRNA during hBMSC chondrogenic differentiation led to significantly increased COL2 and SOX9 expression and decreased COL10 expression. Finally, miR-27b-overexpressing hBMSC chondrogenic pellets had better hyaline cartilage morphology and reduced expression of hypertrophic markers and tend to increase repair efficacy in vivo. Conclusion MiR-27b plays an important role in preventing hypertrophic chondrogenesis of hBMSCs by targeting CBFB and is essential for maintaining a hyaline cartilage state. This study provides new insights into the mechanism of hBMSC chondrocyte differentiation and will aid in the development of strategies for treating cartilage injury based on hBMSC transplantation.

Background The association between pathological tumor size and predictive value in patients with supraglottic carcinoma who receive surgery is poorly understood. To gain further insight into this association, the influence of postoperative tumor size assessed via pathological evaluation on survival outcomes in patients with supraglottic carcinoma was fully analyzed. Materials and methods This study collected data from 375 consecutive supraglottic carcinoma patients who underwent primary surgical treatment between 2005 and 2010. The parameters of tumor size, including tumor diameter, tumor area, and tumor volume, were used to analyze the prognostic abilities of overall survival (OS) and disease-free survival (DFS) in supraglottic carcinoma. Results Twelve women and 363 men were included, with a mean age of 61.2 years. The 5-year OS and DFS rates were 65.4% and 56.2%, respectively. The mean tumor diameter, tumor area, and tumor volume of all patients was 3.1 cm, 8.6 cm2, and 9.1 cm3, respectively. Tumors with higher pT stages had a larger tumor diameter, tumor area, and tumor volume. These three factors were significantly correlated with pT stage, and tumor volume was the strongest factor contributing to pT stage. Patients with a larger tumor diameter, tumor area, and tumor volume had worse OS and DFS. Tumor area and tumor volume were independent prognostic factors of OS and DFS in supraglottic carcinoma patients. Conclusions Patients with a larger tumor size have inferior survival outcomes, and tumor area and tumor volume are independent predictive parameters of survival in supraglottic carcinoma patients who are treated with primary surgery.

OBJECTIVE: The aim of this study was to evaluate the predicting role of tumor volume as evaluated by postoperative pathologic examination on overall survival(OS) and disease free survival(DFS) in patients with local advanced glottic carcinoma. METHODS: In this study, the records of 406 consecutive patients with local advanced glottic carcinoma(T3-T4 stages) who underwent surgery ± chemoradiotherapy from January 2005 to December 2010 were retrospectively reviewed and followed up. The demographic characteristics, disease staging, and pathologic tumor volume were analyzed. The optimal cutoff values of tumor volume for OS and DFS were obtained by using receiver operating characteristic(ROC) curves. The association of tumor volume with T stages were assessed by using Logistic regression model, and the relationship between tumor volume and OS and DFS rates were evaluated by using Cox regression models. RESULTS: The 5-and 10-year OS rates were 62.9% and 55.4%, respectively. The 5-and 10-year DFS rates were 55.5% and 50.8%, respectively. The mean tumor volume was(5.1±6.7) cm(3), T4 stage patients had higher tumor volume than those of patients with T3 stage(P < 0.001). The factor of tumor volume was correlated with T stages by using Logistic regression analyses(OR = 13.81, 95%CI: 6.03-31.59, P < 0.001). The optimal cutoff values of tumor volume that were both at 3 cm(3) for OS and DFS rates were obtained by using ROC curve plots. The OS and DFS rates of glottic carcinoma patients with tumor volume ≤ 3 cm(3) were better when compared with those of patients with tumor volume>3 cm(3)(P < 0.001). Upon multivariate analyses, tumor volume was strongly correlated with poorer OS and DFS rates and remained independent prognostic factors for both the OS and DFS of patients with glottic carcinoma(OS: HR = 1.59, 95%CI: 1.09-2.32, P＝0.017; DFS: HR = 1.54, 95%CI: 1.08-2.20, P = 0.016). CONCLUSION: This study demonstrates that tumor volume is correlated with T stages, and this factor is an independent predictive factor of survival outcomes in patients with local advanced glottic carcinoma.

Primary subglottic carcinoma is rare and surgery is the most common therapeutic strategy for Chinese patients with this disease. To retrospectively evaluate surgically treated primary subglottic carcinoma treated with surgery. Patients with primary subglottic carcinoma who initially underwent surgery from 2005-2010 were grouped by surgical procedures with or without laryngeal function preservation and reviewed. Of 1815 patients with laryngeal cancer, 23 had a subglottic origin. Of these, 21 initially underwent surgery; 12 had 'early' (stage I/II) disease, and nine had 'advanced' (stage III/IV) disease. The actuarial 5-year OS was 73.9% [95% confidence interval (54.1% ∼ 93.7%)] for patients with squamous cell carcinoma. Among patients with early disease, the 5-year OS and DFS were 80% for partial laryngectomy and 71.4% for total laryngectomy. Patients with advanced disease underwent total laryngectomy, and the 5-year OS and DFS were 62.5%. Satisfactory oncologic outcomes can be achieved with initial surgery. Patients with early disease who underwent partial laryngectomy had a comparable prognosis to those who underwent total laryngectomy; deglutition and speech function were maintained.

Background: Preimplantation genetic testing for monogenic disease (PGT-M) has become an effective method for providing couples with the opportunity of a pregnancy with a baby free of spinal muscular atrophy (SMA). Multiple displacement amplification (MDA) overcomes the innate dilemma of very limited genetic material available for PGT-M. Objective: To evaluate the use of MDA, combined with haplotype analysis and mutation amplification, in PGT-M for families with SMA. Methods: MDA was used to amplify the whole genome from single blastomeres or trophectoderm (TE) cells. Exon 7 of the survival motor neuron gene 1 (SMN1) and eleven STRs markers flanking the SMN1 gene were incorporated into singleplex polymerase chain reaction (PCR) assays on MDA products. Results: Sixteen cycles (19 ovum pick-up cycles) of PGT-M were initiated in 12 couples. A total of 141 embryos were tested: 90 embryos were biopsied at the cleavage stage and 51 embryos were biopsied at the blastocyst stage. MDA was successful on 94.44% (85/90) of the single blastomeres and on 92.16% (47/51) of the TE cells. And the PCR efficiency were 98.4% (561/570) and 100% (182/182), respectively. In addition, the average allele drop-out (ADO) rates were 13.3% (60/392) and 9.8% (11/112), respectively. The results for SMN1 exon 7 were all matched with haplotype analysis, which allowed an accurate diagnosis of 93.62% (132/141) embryos. Twelve families had unaffected embryos available for transfer and a total of 38 embryos were transferred in 20 embryo transfer cycles. Eight transfers were successful, resulting in a clinical pregnancy rate of 40% (8/20) and an implantation rate of 28.95% (11/38). Finally, 11 healthy babies were born. Among them, 5 SMA carriers were singleton live births and 3 SMA carriers had twin births. Conclusion: Careful handling during the MDA procedure can improve subsequent PCR efficiency and reduce the ADO rate. We suggest that this protocol is reliable for increasing the accuracy of the PGT-M for SMA.

Objective: To investigate the association between the cytokeratin (CK)-1 single-nucleotide polymorphism (SNP), the protein level of CK-1 and the risk of vocal leukoplakia and laryngeal squamous cell carcinoma (LSCC). Methods: In this case-control study, 155 patients with vocal leukoplakia, 323 patients with LSCC, and 266 healthy controls were genotyped for the CK-1 (SNP RS14024) gene using pyrosequencing. The protein expression level of CK-1 was analyzed in vocal leukoplakia, LSCC, and vocal polyp patients by immunohistochemistry (IHC). Results: Of the CK-1 RS14024 polymorphism, the heterozygote AG and homozygote GG genotype exhibited a significantly increased risk of LSCC (AG: OR = 2.16, p = 0.014; GG: OR = 2.15, p = 0.018) compared to normal controls. A higher protein expression level of CK-1 was detected in patients with LSCC compared to vocal leukoplakia and polyps (both p < 0.001), and a significant increasing trend of CK-1 protein expression level from mild-moderate dysplasia to moderate-severe dysplasia in vocal leukoplakia patients was also observed (p = 0.006). Conclusions: This study demonstrates that the CK-1 SNP and high protein expression levels are associated with vocal leukoplakia and LSCC and promote the transformation from vocal leukoplakia to LSCC in a Chinese Han population.

Preoperative tracheotomy is an effective option that secures upper airway patency in laryngeal carcinoma patients suffering from upper airway obstruction, but the influence of this treatment on oncologic outcomes of laryngeal carcinoma remains controversial. The purpose of this study was to determine the impact of preoperative tracheotomy on overall survival in supraglottic carcinoma patients with tumor obstruction of the upper airway, and explore the potential causes.This retrospective study collected 243 consecutive patients with advanced stage supraglottic carcinoma from 2005 to 2010. Preoperative tracheotomy in the management of upper airway obstruction in patients with supraglottic carcinoma was analyzed.The mean age was 60.9 years at diagnosis, with men accounting for 98.4% of all patients. Thirty nine (16.0%) patients presenting with tumor obstruction of the upper airway required preoperative tracheotomy. T4 stage patients had higher rate of tracheotomy than those of patients with T3 stage (36.8% vs 12.2%). Patients with upper airway obstruction presented with greater tumor area compared with patients without (13.7 cmAdvanced stage supraglottic carcinoma patients with upper airway obstruction undergoing preoperative tracheotomy experienced worse overall survival. Advanced T stage and greater tumor size were associated with upper airway obstruction, indicating that the negative influence of tumor obstruction on survival may be cause by these two preoperative variables. Therefore, preoperative tracheotomy acts only as an alternative procedure, and is not a prognostic agent.

An excessive RF power requirement is one of the main obstacles in the clinical translation of EPR imaging. The radio frequency (RF) pulses used in EPR imaging to excite electron spins must be very short to match their fast relaxation. With traditional pulse schemes and ninety degree flip angles, this can lead to either unsafe specific absorption rate (SAR) levels or unfeasibly long repetition times. In spectroscopy experiments, it has been shown that stochastic excitation and correlation detection can reduce the power while maintaining sensitivity but have yet to be applied to imaging experiments. Stochastic excitation is implemented using a pseudo-random phase modulation of the input stimulus. Using a crossed coil resonator assembly comprised of an outer saddle coil and an inner surface coil, it was possible to obtain a minimum isolation of ∼50 dB across a 12 MHz bandwidth. An incident peak RF power of 5 mW was used to excite the system. The low background signal obtained from this resonator allowed us to generate images with 32 dB (>1000:1) signal-to-noise ratio (SNR) while exciting with a traditional pulse sequence in a phantom containing the solid paramagnetic probe NMP-TCNQ (N-methyl pyridinium tetracyanoquinodimethane). Using two different stochastic excitation schemes, we were able to achieve a greater than 4-fold increase in SNR at the same peak power and number of averages, compared to single pulse excitation. This procedure allowed imaging at significantly lower RF power levels than used in conventional EPR imaging system configurations. Similar techniques may enable clinical applications for EPR imaging by facilitating the use of larger RF coils while maintaining a safe SAR level.

T3 supraglottic laryngeal carcinoma (LC) is a common advanced laryngeal cancer.This study was conducted to assess the clinical results of pathological T3 (pT3) supraglottic LC patients who were amenable to laryngectomy treated with primary surgery and postoperative therapy.Retrospective review of 202 pT3 cases of supraglottic laryngeal squamous cell carcinoma.The five-year cancer specific survival (CSS) rate was 63.7% and the overall survival rate (OS) was 62.8%. For T3 supraglottic patients who underwent total laryngectomy, the five-year disease-free survival (DFS) was 51.8%, and the CSS was 62.5%. For patients who underwent partial laryngectomy, the five-year DFS was 72.2%, and the CSS was 79.0%. High lymph node and stage status are predictors of mortality for these patients. No difference was found in the DFS and CSS rates between patients with negative margins and those with positive margins following postoperative radiotherapy and chemotherapy.Surgical treatment of T3 supraglottic LC patients achieved satisfactory results. Postoperative radiotherapy and chemotherapy are an effective method of treatment for T3 supraglottic LC patients, especially for those with a positive margin.

Background To investigate the therapeutic mechanism of the BRD4 inhibitor JQ1 in SACC-83 cells and explore strategies to enhance its therapeutic potential. Material and methods SACC-83 cells were used in the experiment. Immunohistochemistry was used to assess BRD4 expression in SACC tissues and corresponding adjacent non-tumor tissues. Cell viability and proliferation were evaluated using the Cell Counting Kit-8 assay. Flow cytometry was used to quantitate apoptosis. Levels of cleaved caspase-3, BRD4, c-Myc, pEGFR (γ-1173), and EGFR were determined by quantitative real-time PCR and Western blot. To study the role of EGFR in JQ1 resistance, we generated EGFR knockdown SACC-83 cells by siRNA transfection. Results Our study revealed that BRD4 was overexpressed and could be a treatment target in SACC. The BRD4 inhibitor JQ1 markedly inhibited c-Myc expression in SACC-83 cells, which produced modest therapeutic effects. Nevertheless, the EGFR pathway was strongly activated following JQ1 treatment, which led to JQ1 resistance. Combined JQ1 and PI3K inhibitor treatment effectively increased the therapeutic potential by inhibiting the EGFR and c-Myc signaling pathways in SACC-83 cells. Moreover, EGFR knockdown sensitized SACC-83 cells to JQ1. Conclusion These data demonstrate that EGFR and c-Myc signaling synergistically drive SACC progression. The JQ1 and PI3K inhibitor combination exhibited a strong synergistic effect by suppressing c-Myc and EGFR in SACC-83 cells, identifying a novel rational combinatorial treatment. Moreover, EGFR expression influences the sensitivity of SACC-83 cells to JQ1, which is useful for planning treatment.

Inhibiting BRD4 has emerged as a promising anticancer strategy, and inhibitors such as JQ1 can suppress cell growth in oral squamous cell carcinoma (OSCC). However, the mechanism through which JQ1 exerts its anticancer activity has not been reported. Moreover, JQ1 does not markedly inhibit proliferation and increase apoptosis in OSCC when used as a monotherapy. Herein, we explore the mechanism of JQ1 in OSCC and probe ways to increase its therapeutic potential. In this study, we used two cell lines, Cal27, and Scc25. We found that BRD4 was highly expressed in OSCC tissues when compared with adjacent non-tumor tissues, and JQ1 worked through the EGFR-mediated signaling pathway in tumor cells. Furthermore, we demonstrated that JQ1 induced an increased treatment effect in vitro and in vivo when combined with a PI3K inhibitor. Interestingly, subsequent mechanistic analyses indicated that further suppressing EGFR and BRD4 expression was instrumental to this functional synergism. Moreover, we found that upregulating EGFR expression by EGF stimulation protected cells treated with JQ1 from apoptosis, while knockdown of EGFR before addition of JQ1 successfully mimicked the combination treatment results. In summary, our findings revealed that JQ1 can act by inhibiting the EGFR-mediated signaling pathway, and EGFR expression influences the sensitivity of OSCC to JQ1. Regarding clinical use, this study demonstrates that BRD4 is a novel therapeutic target and EGFR can be used as a biomarker to identify the most appropriate anti-BRD4 treatment strategy in OSCC.

PURPOSE: This paper aims to investigate the feasibility of performing pre-implantation genetic diagnosis (PGD) and pre-implantation genetic screening (PGS) simultaneously by a universal strategy without the requirement of genotyping relevant affected family members or lengthy preliminary work on linkage analysis. METHODS: By utilizing a universal Mutated Allele Revealed by Sequencing with Aneuploidy and Linkage Analyses (MARSALA) strategy based on low depth whole genome sequencing (~3x), not involving specific primers' design nor the enrichment of SNP markers for haplotype construction. Single-sperm cells and trephectoderm cells from in vitro fertilized embryos from a couple carrying HBB mutations were genotyped. Haplotypes of paternal alleles were constructed and investigated in embryos, and the chromosome copy number profiles were simultaneously analyzed. RESULTS: The universal MARSALA strategy allows the selection of a euploid embryo free of disease mutations for in uterus transfer and successful pregnancy. A follow-up amniocentesis was performed at 17 weeks of gestation to confirm the PGD/PGS results. CONCLUSION: We present the first successful PGD procedure based on genotyping multiple single-sperm cells to obtain SNP linkage information. Our improved PGD/PGS procedure does not require genotyping the proband or relevant family members and therefore can be applicable to a wider population of patients when conducting PGD for monogenic disorders.

It has been demonstrated that mesenchymal stem cells (MSCs) have potent immunosuppressive capacities. But it is controversial whether differentiated mature stromal cells (SCs) share the immunosuppressive capacities. A previous study examined the ability of SCs from different human tissue sites to inhibit the proliferation of lymphocytes. The results are all positive but the mechanism isn't clear, and few mouse data have been published on this topic. Using an efficient mixed cell culture assay, our in vitro data show that the anti-proliferative ability of murine MSCs on lymphocytes is shared by mature murine SCs, i.e. chondrocytes and fibroblasts. Though conflicting results have been published, our results suggest that nitric oxide and IFN-γ are critical to the immunosuppressive effect. We also demonstrate that murine MSCs cultivated in chondrogenic differentiation medium still possess the anti-proliferative capacities on lymphocytes in vitro.

Astrocytes control many processes of the nervous system in health and disease, and respond to injury quickly. Astrocytes produce neuroprotective factors in the injured brain to clear cellular debris and to orchestrate neurorestorative processes that are beneficial for neurological recovery after traumatic brain injury (TBI). However, astrocytes also become dysregulated and produce cytotoxic mediators that hinder CNS repair by induction of neuronal dysfunction and cell death. Hence, we discuss the potential role of astrocytes in neuropathological processes such as neuroinflammation, neurogenesis, synaptogenesis and blood-brain barrier repair after TBI. Thus, an improved understanding of the dual role of astrocytes may advance our knowledge of post-brain injury recovery, and provide opportunities for the development of novel therapeutic strategies for TBI.