Biomaterial-Based CRISPR/Cas9 Delivery Systems for Tumor Treatment.

CRISPR/Cas9 gene editing technology is characterized by high specificity and efficiency, and has been applied to the treatment of human diseases, especially tumors involving multiple genetic modifications. However, the clinical application of CRISPR/Cas9 still faces some major challenges, the most urgent of which is the development of optimized delivery vectors. Biomaterials are currently the best choice for use in CRISPR/Cas9 delivery vectors owing to their tunability, biocompatibility, and efficiency. As research on biomaterial vectors continues to progress, hope for the application of the CRISPR/Cas9 system for clinical oncology therapy builds. In this review, we first detail the CRISPR/Cas9 system and its potential applications in tumor therapy. Then, we introduce the different delivery forms and compare the physical, viral, and non-viral vectors. In addition, we analyze the characteristics of different types of biomaterial vectors. We further review recent research progress in the use of biomaterials as vectors for CRISPR/Cas9 delivery to treat specific tumors. Finally, we summarize the shortcomings and prospects of biomaterial-based CRISPR/Cas9 delivery systems.

Pulmonary inhalation for disease treatment: Basic research and clinical translations.

Pulmonary drug delivery has the advantages of being rapid, efficient, and well-targeted, with few systemic side effects. In addition, it is non-invasive and has good patient compliance, making it a highly promising drug delivery mode. However, there have been limited studies on drug delivery via pulmonary inhalation compared with oral and intravenous modes. This paper summarizes the basic research and clinical translation of pulmonary inhalation drug delivery for the treatment of diseases and provides insights into the latest advances in pulmonary drug delivery. The paper discusses the processing methods for pulmonary drug delivery, drug carriers (with a focus on various types of nanoparticles), delivery devices, and applications in pulmonary diseases and treatment of systemic diseases (e.g., COVID-19, inhaled vaccines, diagnosis of the diseases, and diabetes mellitus) with an updated summary of recent research advances. Furthermore, this paper describes the applications and recent progress in pulmonary drug delivery for lung diseases and expands the use of pulmonary drugs for other systemic diseases.

Flourishing tumor organoids: History, emerging technology, and application.

Malignant tumors are one of the leading causes of death which impose an increasingly heavy burden on all countries. Therefore, the establishment of research models that closely resemble original tumor characteristics is crucial to further understanding the mechanisms of malignant tumor development, developing safer and more effective drugs, and formulating personalized treatment plans. Recently, organoids have been widely used in tumor research owing to their advantages including preserving the structure, heterogeneity, and cellular functions of the original tumor, together with the ease of manipulation. This review describes the history and characteristics of tumor organoids and the synergistic combination of three-dimensional (3D) culture approaches for tumor organoids with emerging technologies, including tissue-engineered cell scaffolds, microfluidic devices, 3D bioprinting, rotating wall vessels, and clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 (CRISPR-Cas9). Additionally, the progress in research and the applications in basic and clinical research of tumor organoid models are summarized. This includes studies of the mechanism of tumor development, drug development and screening, precision medicine, immunotherapy, and simulation of the tumor microenvironment. Finally, the existing shortcomings of tumor organoids and possible future directions are discussed.

cdh23 affects congenital hearing loss through regulating purine metabolism.

Introduction: The pathogenic gene CDH23 plays a pivotal role in tip links, which is indispensable for mechanoelectrical transduction in the hair cells. However, the underlying molecular mechanism and signal regulatory networks that influence deafness is still largely unknown. Methods: In this study, a congenital deafness family, whole exome sequencing revealed a new mutation in the pathogenic gene CDH23, subsequently; the mutation has been validated using Sanger sequencing method. Then CRISPR/Cas9 technology was employed to knockout zebrafish cdh23 gene. Startle response experiment was used to compare with wide-type, the response to sound stimulation between wide-type and cdh23-/-. To further illustrate the molecular mechanisms underlying congenital deafness, comparative transcriptomic profiling and multiple bioinformatics analyses were performed. Results: The YO-PRO-1 assay result showed that in cdh23 deficient embryos, the YO-PRO-1 signal in inner ear and lateral line neuromast hair cells were completely lost. Startle response experiment showed that compared with wide-type, the response to sound stimulation decreased significantly in cdh23 mutant larvae. Comparative transcriptomic showed that the candidate genes such as atp1b2b and myof could affect hearing by regulating ATP production and purine metabolism in a synergetic way with cdh23. RT-qPCR results further confirmed the transcriptomics results. Further compensatory experiment showed that ATP treated cdh23-/- embryos can partially recover the mutant phenotype. Conclusion: In conclusion, our study may shed light on deciphering the principal mechanism and provide a potential therapeutic method for congenital hearing loss under the condition of CDH23 mutation.

Effects of modified posterior nasal nerve neurectomy combined with accessory posterior nasal nerve neurectomy on controlling intractable allergic rhinitis. / 改良鼻后神经切断术联合副鼻后神经切断术治疗难治型变应性鼻炎的效果（英文）.

OBJECTIVES: A subset of intractable allergic rhinitis (iAR) patients experience severe symptoms which cannot be effectively controlled by standard drug therapy and/or antigen specific immunotherapy. In recent decades, endoscopy vidian neurectomy and posterior nasal nerve neurectomy (PNNN) were introduced as treatments of iAR that have shown to be highly successful at symptom management in a number of patients. But some patients experience relapse or suboptimal symptom control postoperation. To improve the effectiveness of PNNN to control iAR, a modified PNNN surgical approach (mPNNN) combined with accessory posterior nasal nerve neurectomy (aPNNN), which called as mPNNN-aPNNN was used. This study aims to compare the effects between mPNNN-aPNNN and PNNN on controlling the symptoms of iAR and evaluate the surgical effectiveness and safety of mPNNN-aPNNN. METHODS: The patients with iAR experienced mPNNN-aPNNN or PNNN surgery at the department of Otolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were analyzed retrospectively. The approach of PNNN, a selective resection of the posterior nasal nerve branches, was modified to the neurectomy of total branches of posterior nasal nerve at the sphenopalatine foramen, and combined the operation of aPNNN in which the accessory posterior nasal nerve at the palatine bone perpendicular plate was resect in our study. Daily Nasal Symptom Scores (DNSS), Total Rhinitis Medication Score (TRMS), and the Rhinoconjunctivitis Qualities of Life Questionnaires Scores (RQLQS) were used to evaluate the complications during the operation and after the operation at the 3rd, 6th, 12th, and 24th month postoperatively. Total Nasal Symptom Scores (TNSS) was used to assess the total effective rate and markedly effective rate of the operations. RESULTS: A total of 140 iAR patients experienced mPNNN-aPNNN or PNNN. Those with concomitant septoplasty and/or inferior turbinate reduction, and were absent during the postoperative follow-up were excluded. The final 62 patients with mPNNN-aPNNN and 34 with PNNN were enrolled. DNSS, TNSS, TRMS, and RQLQS at the postoperation were significantly improved compared with the preoperation in all patients (all P<0.001). Compared with PNNN, the postoperative DNSS, TNSS, and TRMS of mPNNN-aPNNN were obviously improved (all P<0.001). There was a persisted relief of symptoms at the postoperation in all patients with mPNNN-aPNNN. The total effective rate and markedly effective rate at the postoperative 24th month were 100% and 83.3%, respectively. Furthermore, the postoperative RQLQS decreased significantly (P<0.001). Only 5 sides of all patients (5/192, 2.6%) reported upper palate numbness during the first week after operation, with all recovered spontaneously in 1 month without treatment. No other postoperative complications occurred in mPNNN-aPNNN and PNNN. CONCLUSIONS: The surgery of mPNNN-aPNNN improve TNSS more significantly than PNNN. The operation of mPNNN-aPNNN is safe and effective to control iAR symptoms.

Functional acellular matrix for tissue repair.

In view of their low immunogenicity, biomimetic internal environment, tissue- and organ-like physicochemical properties, and functionalization potential, decellularized extracellular matrix (dECM) materials attract considerable attention and are widely used in tissue engineering. This review describes the composition of extracellular matrices and their role in stem-cell differentiation, discusses the advantages and disadvantages of existing decellularization techniques, and presents methods for the functionalization and characterization of decellularized scaffolds. In addition, we discuss progress in the use of dECMs for cartilage, skin, nerve, and muscle repair and the transplantation or regeneration of different whole organs (e.g., kidneys, liver, uterus, lungs, and heart), summarize the shortcomings of using dECMs for tissue and organ repair after refunctionalization, and examine the corresponding future prospects. Thus, the present review helps to further systematize the application of functionalized dECMs in tissue/organ transplantation and keep researchers up to date on recent progress in dECM usage.

Preservation of superior laryngeal nerve in transoral surgery: A technology to enhance the recovery of swallowing function after surgery of hypopharyngeal carcinoma.

OBJECTIVES: We intended to preserve the internal branch of superior laryngeal nerve in transoral surgery of hypopharyngeal squamous cell carcinoma and observe swallowing function recovery. METHODS: 26 patients with hypopharyngeal squamous cell carcinoma underwent transoral surgery with the preservation of internal branch of superior laryngeal nerve. Sensation in the pharyngolaryngeal mucosa was tested by flexible laryngoscope and swallow function was evaluated by water swallow test and MD Anderson Dysphagia Inventory questionnaire after surgery. RESULTS: Surgeries were successfully performed in all patients. The internal branch of superior laryngeal nerve were preserved in all patients. Testing of mucosa sensation revealed the presence of the cough reflex in most patients. The water swallow test showed that 12 cases (46.15%) on the 1st day, 23 cases (88.46%) on the 7th day and 25 cases (96.15%) on the 14th day after operation had normal swallowing function. The mean score of MD Anderson Dysphagia Inventory was 98 on the 14th day after operation. All patients achieved an oral soft diet at a median of 3 days (range, 2-6 days), full normal oral diet at a median of 5.5 days (range, 4-10 days) and removal of the nasogastric tube at a median of 6 days (range, 5-11 days). During the two-year follow-up, 3 patients recured, 1 patient died of lung metastasis. CONCLUSIONS: Preserving of the internal branch of superior laryngeal nerve in transoral surgery is feasible, and it can help to achieve a satisfactory recovery of the swallowing function after surgery of hypopharyngeal squamous cell carcinoma.

FN1 promotes prognosis and radioresistance in head and neck squamous cell carcinoma: From radioresistant HNSCC cell line to integrated bioinformatics methods.

Background: Radioresistance in head and neck squamous cell carcinoma (HNSCC) patients means response failure to current treatment. In order to screen radioresistant biomarkers and mechanisms associated with HNSCC, differentially expressed genes (DEGs) associated with radioresistance in HNSCC were investigated. Methods: The HNSCC cell line with radioresistance, Hep2-R, was established and detected the radiosensitivity using MTT, colony formation assay and flow cytometry analysis. Clariom™ D chip was applied to compare DEGs between Hep2 and Hep2-R groups and build the differential gene expression profiles associated with radioresistance in HNSCC. Bioinformatic analysis were used to find biological functions and pathways that related to radioresistance in HNSCC, including cell adhesion, cytochrome P450 and drug metabolism. Gene Expression Omnibus (GEO) datasets were selected to verify DEGs between HNSCC radioresistant cells and tissues. The representation of DEGs were validated between HNSCC patients with complete response and post-operative radiation therapy failure. In addition, we evaluated the clinical prognosis of DEGs using The Cancer Genome Atlas (TCGA) database. Results: 2,360 DEGs (|Fold Change|>1.5, p < 0.05) were identified between Hep2 and Hep2-R, including 1,144 upregulated DEGs and 1,216 downregulated DEGs. They were further verified by HNSCC radioresistant cells and tissues in GEO. 13 radioresistant DEGs showed same difference in expression level between cells and tissues. By comparing 13 DEGs with HNSCC patients, upregulations of FN1, SOX4 and ETV5 were found identical with above results. Only FN1 was a prognostic indicator of HNSCC in TCGA. Conclusion: FN1 is the potential novel biomarker for predicting poor prognosis and radioresistance in HNSCC patients. Overexpression of FN1 plays an important role in the tumorigenesis, prognosis and radioresistance of HNSCC.

Biometrics-protected optical communication enabled by deep learning-enhanced triboelectric/photonic synergistic interface.

Security is a prevailing concern in communication as conventional encryption methods are challenged by progressively powerful supercomputers. Here, we show that biometrics-protected optical communication can be constructed by synergizing triboelectric and nanophotonic technology. The synergy enables the loading of biometric information into the optical domain and the multiplexing of digital and biometric information at zero power consumption. The multiplexing process seals digital signals with a biometric envelope to avoid disrupting the original high-speed digital information and enhance the complexity of transmitted information. The system can perform demultiplexing, recover high-speed digital information, and implement deep learning to identify 15 users with around 95% accuracy, irrespective of biometric information data types (electrical, optical, or demultiplexed optical). Secure communication between users and the cloud is established after user identification for document exchange and smart home control. Through integrating triboelectric and photonics technology, our system provides a low-cost, easy-to-access, and ubiquitous solution for secure communication.

Congenital Middle Ear Malformation with Common Deafness Gene Mutation Analysis: A Review of 813 Profound Sensorineural Hearing Loss Child Patients.

Deafness gene variants play a key role in inner ear malformations. However, the relationship between congenital middle ear malformations and common deafness genes (GJB2, SLC26A4, and mtDNA) in profound sensorineural hearing loss (SNHL) child patients remains poorly investigated. Here we showed that there was no statistical significance in the total mutation frequency of the three common deafness genes in the middle ear malformation group (21.2%, 41/193) in comparison with the normal middle ear and inner ear group (21.0%, 116/553) (χ2 = 0.0061, p = 0.940). Moreover, the mutation ratio of GJB2 and SLC26A4 in the middle ear malformation group (18.7%, 36/193; 2.6%, 5/193) was not significantly different from that in the normal middle ear and inner ear group (17.7%, 98/553; 2.4%, 13/553) (χ2 = 0.084, p = 0.772; χ2 = 0.0000, p = 1.000). The mutation ratio of GJB2 235delC and GJB2 79G>A in the middle ear malformation group (8.8%, 17/193; 8.8%, 17/193) was almost the same to that in the normal middle ear and inner ear group (8.6%, 48/553; 6.7%, 37/553) (χ2 = 0.0030, p = 0.957; χ2 = 0.9556, p = 0.328). The high jugular bulb subgroup analysis also showed the same results. Our findings suggested that GJB2, SLC26A4, and mtDNA mutations might not be related to the middle ear malformations in profound SNHL child patients. Anat Rec, 303:594-599, 2020. © 2019 American Association for Anatomy.

TGF-ß1 Induces Epithelial-Mesenchymal Transition of Chronic Sinusitis with Nasal Polyps through MicroRNA-21.

OBJECTIVES: To characterize the epithelial-mesenchymal transition (EMT) in chronic rhinosinusitis with nasal polyps (CRSwNP) and to investigate the mechanism by which microRNA-21 (miR-21) regulates EMT in CRSwNP. METHOD: (1) Tissue experiments: Mucosa tissues were collected from 13 patients with CRSwNP and 12 patients with CRS without nasal polyps (CRSsNP), as well as 11 patients without CRS (controls). Protein localization and quantification were achieved by immunofluorescence staining and Western blotting, involving the epithelial marker protein E-cadherin and the mesenchymal marker proteins α-smooth muscle actin (α-SMA), fibronectin, and vimentin. Quantitative RT-PCR was used to detect the relative expression levels of miR-21 and TGF-ß1 mRNAs. (2) Cellular experiments: Primary human nasal epithelial cells (PHNECs) treated with TGF-ß1, or TGF-ß1 with miR-21 inhibitor, or miR-21 mimics alone were observed for morphology changes under a phase-contrast microscope. The expression levels of epithelial/mesenchymal marker proteins were determined as aforementioned. PTEN and phosphorylated Akt were detected by Western blotting. RESULTS: (1) Tissue experiments: Compared with the CRSsNP and control groups, the expression of E-cadherin was downregulated in the CRSwNP group, whereas the expression of TGF-ß1, α-SMA, fibronectin, and vimentin was upregulated. The expression levels of miR-21 and TGF-ß1 mRNAs in CRSwNP were significantly higher than those in CRSsNP and controls. (2) Cellular experiments: TGF-ß1 induced EMT-like transformation in PHNECs, featured by changes in cell morphology and upregulation of mesenchymal proteins and miR-21. The miR-21 inhibitor, as well as the Akt-specific -inhibitor, suppressed TGF-ß1-induced EMT. Mechanically, downregulation of miR-21 resulted in increased PTEN and decreased Akt phosphorylation. Furthermore, overexpression of miR-21 had the opposite effects. CONCLUSIONS: Our findings suggest that the TGF-ß1-miR-21-PTEN-Akt axis may contribute to the pathogenesis of CRSwNP. miR-21 might be a reliable target for treating nasal polyp genesis through EMT suppression. Moreover, miR-21 inhibitors could be a novel class of antipolyp drug that modulates PTEN expression and Akt activation. In addition, further investigation regarding the reason underlying miR-21 overexpression in CRSwNP could provide a molecular target for novel treatment strategies for nasal polyposis.

Anti-neutrophil cytoplasmic antibody-associated hypertrophic cranial pachymeningitis and otitis media: a review of literature.

BACKGROUND AND OBJECTIVE: It has been recognized that anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides may lead to hypertrophic pachymeningitis (HP) or intractable otitis media (OM). To our knowledge, few cases of coexistent ANCA-related HP and OM have been described previously. To increase awareness of this disease, we reviewed the literature describing patients with HP and intractable OM in a population with AAV to guide clinical decision making for otolaryngologists. METHODS:  PubMed was searched with the following terms: ANCA-associated vasculitis, otitis media, and hypertrophic pachymeningitis. Only patients with concomitant AAV, OM and HP were considered and included in this review. RESULTS: A total of 243 articles were reviewed, and of these, 6 met inclusion criteria. Headache, cranial polyneuropathy, and intractable OM with effusion or granulation were common. Serum MPO-ANCA positivity was most common in Asian patients. Almost all patients had dural mater thickening on gadolinium-enhanced magnetic resonance imaging of the brain. Corticosteroids plus an immunosuppressant was more effective and most patients had improved hearing after treatment, but approximately 50% of subjects had disease relapse. CONCLUSION: In this review, we summarized the current knowledge on the clinical features, diagnosis, treatment, and pathogenesis of this disease. We should carefully detect the potential cases of ANCA-related HP and OM in patients with intractable OM, HP, or AAV, and make the optimal treatment plan to avoid long-term neurological complications and irreversible hearing loss. Furthermore, due to an increased possibility of relapse, close follow-up, including a hearing test, ANCA titers, imaging examination, and detection of toxic and side effects of immunosuppressive therapy, are necessary.

Intelligence development of pre-lingual deaf children with unilateral cochlear implantation.

OBJECTIVE: The present study aims to test whether deaf children with unilateral cochlear implantation (CI) have higher intelligence quotients (IQ). We also try to find out the predictive factors of intelligence development in deaf children with CI. METHODS: Totally, 186 children were enrolled into this study. They were divided into 3 groups: CI group (N = 66), hearing loss group (N = 54) and normal hearing group (N = 66). All children took the Hiskey-Nebraska Test of Learning Aptitude to assess the IQ. After that, we used Deafness gene chip, Categories of Auditory Performance (CAP) and Speech Intelligibility Rating (SIR) methods to evaluate the genotype, auditory and speech performance, respectively. RESULTS: At baseline, the average IQ of hearing loss group (HL), CI group, normal hearing (NH) group were 98.3 ± 9.23, 100.03 ± 12.13 and 109.89 ± 10.56, while NH group scored higher significantly than HL and CI groups (p < 0.05). After 12 months, the average IQ of HL group, CI group, NH group were99.54 ± 9.38,111.85 ± 15.38, and 112.08 ± 8.51, respectively. No significant difference between the IQ of the CI and NH groups was found (p > 0.05). The growth of SIR was positive correlated with the growth of IQ (r = 0.247, p = 0.046), while no significant correlation were found between IQ growth and other possible factors, i.e. gender, age of CI, use of hearing aid, genotype, implant device type, inner ear malformation and CAP growth (p > 0.05). CONCLUSIONS: Our study suggests that CI potentially improves the intelligence development in deaf children. Speech performance growth is significantly correlated with IQ growth of CI children. Deaf children accepted CI before 6 years can achieve a satisfying and undifferentiated short-term (12 months) development of intelligence.

Progressive hearing loss and degeneration of hair cell stereocilia in taperin gene knockout mice.

The TPRN gene encodes taperin, which is prominently present at the taper region of hair cell stereocilia. Mutations in TPRN have been reported to cause autosomal recessive nonsyndromic deafness 79(DFNB 79). To investigate the role of taperin in pathogenesis of hearing loss, we generated TPRN knockout mice using TALEN technique. Sanger sequencing confirmed an 11 bp deletion at nucleotide 177-187 in exon 1 of TPRN, which results in a truncated form of taperin protein. Heterozygous TPRN+/- mice showed apparently normal auditory phenotypes to their wide-type (WT) littermates. Homozygous TPRN-/- mice exhibited progressive sensorineural hearing loss as reflected by auditory brainstem response to both click and tone burst stimuli at postnatal days 15 (P15), 30 (P30), and 60 (P60). Alex Fluor-594 phalloidin labeling showed no obvious difference in hair cell numbers in the cochlea between TPRN-/- mice and WT mice under light microscope. However, scanning electronic microscopy revealed progressive degeneration of inner hair cell stereocilia, from apparently normal at postnatal days 3 (P3) to scattered absence at P15 and further to substantial loss at P30. The outer hair cell stereocilia also showed progressive degeneration, though much less severe, Collectively, we conclude that taperin plays an important role in maintenance of hair cell stereocilia. Establishment of TPRN knockout mice enables further investigation into the function of this gene.

Imaging assessment of profound sensorineural deafness with inner ear anatomical abnormalities.

OBJECTIVE: To explore the value of a combined computed tomography (CT) and magnetic resonance imaging (MRI) in evaluating profound sensorineural deafness patients before cochlear implant (CI) surgery. METHODS: A retrospective analysis of 1012 cases of profound sensorineural deafness that received CI was performed. RESULTS: A total of 96 cases were diagnosed with inner ear abnormalities including large vestibular aqueduct syndrome (LVAS, n = 61), Michel deformity (n = 3), cochlear incomplete partition I (n = 2), cochlear incomplete partition II (n = 6), cochlear hypoplasia with vestibular malformation (n = 3), cochlear ossification (n = 3), bilateral internal auditory canal obstruction (n = 5) and internal auditory canal stenosis (n = 2). CONCLUSION: High resolution CT (HRCT) can display bony structures while MRI can image the membranous labyrinth in preoperative evaluation for cochlear implantation. The combination of these two modalities provides reliable anatomical information regarding the bony and membranous labyrinths, as well as the auditory nerve.

Prevalence of mutations in GJB2, SLC26A4, and mtDNA in children with severe or profound sensorineural hearing loss in southwestern China.

AIM: To study the distribution characteristics of common mutations in the GJB2, SLC26A4, and mtDNA genes in children with severe or profound sensorineural hearing loss (SNHL) in southwestern China. MATERIALS AND METHODS: A total of 1,164 individuals were recruited to screen for the common GJB2, SLC26A4, and mtDNA mutations by microarrays. Subsequencing for the coding region of the GJB2 gene in the samples without the GJB2 hotspot mutations as well as subsequencing for the exon 1 of the TRMU gene in those samples with the mtDNA hotspot mutations was performed by Sanger sequencing. All mutations were analyzed in association with medical imaging. RESULTS: In this study, 28.43% of all subjects carried mutations. The mutation frequencies in the GJB2, SLC26A4, and mtDNA genes were 17.27%, 7.04%, and 4.12%, respectively. No TRMU mutation was found in the study. The frequency of the mtDNA mutations in the multiethnic minorities was six times that in the Han (11.23% vs. 1.91%; p approaches 0.000) and in the urban group was one-third of that in the suburban group(1.49% vs. 4.47%; p=0.047). The frequency of the GJB2 mutations in urban and suburban groups was 23.38% and 15.99%, respectively (p=0.012). The enlarged vestibular aqueduct (EVA) was the most common inner ear malformation and ∼79.10% of EVA cases were associated with the SLC26A4 mutations. CONCLUSIONS: More than one-fourth of children with severe or profound SNHL carried the common deafness mutations. The proportions of ethnic minorities and urban subjects could impact the frequency of the GJB2 and mtDNA mutations. The SLC26A4 hotspot mutations are prevalent and correlate strongly with EVA.

A review of the management and prognosis of thyroid carcinoma with tracheal invasion.

The objective of the study was to explore the surgical approaches, treatment significance and prognosis of thyroid carcinoma with tracheal invasion. We retrospectively reviewed 48 patients with tracheal invasion by papillary thyroid carcinoma, follicular thyroid carcinoma and medullary thyroid carcinoma by means of clinical data ranging from 1993 to 2011. The patients were classified into three groups in terms of the depth and extent of tracheal invasion by the tumors, i.e., group A of 18 patients with localized tracheal invasion; group B of 21 patients with intraluminal tracheal invasion, and group C of nine patients with extensive invasion of the trachea, larynx, esophagus and/or mediastinum. Of these patients, 18 received radical tumorectomy and segmental resection of the outer layer of the tracheal wall; 21 had radical tumorectomy, circumferential sleeve trachea resection or segmental tracheal resection plus tracheal repair, and the remaining nine patients underwent radical tumorectomy, segmental trachea resection and esophagolaryngectomy. 46 patients took I(131) oral solution and/or had external radiotherapy postoperatively. A survival analysis was done using Kaplan-Meier Estimator for cumulative survival probability together with Log-Rank test, and Cox Regression Model was used for multivariate analysis. (1) In group A of 18 patients, 10 took I(131)oral solution and 7 received radiotherapy after surgery. The overall 5 and 10-year survival rates were 88.93 and 77.78 %, respectively; (2) In group B of 21 patients, 15 took I(131) oral solution and 7 received radiotherapy after surgery. The overall 5 and 10-year survival rates were 90.47 and 61.87 %, respectively; (3) In group C of 9 patients, 7 received radiotherapy after surgery. The overall 5 and 10-year survival rates were 77.78 and 22.22 %, respectively. Whether they received postsurgical I(131) treatment or radiotherapy, there was a statistical difference between the 5-year survival rates and the 10-year survival rates in all of these three groups (P value in each group is <0.05). In the treatment of thyroid carcinoma with tracheal invasion, radical tumorectomy plus tracheal repair, segmental tracheal resection or circumferential sleeve trachea resection could lengthen the survival time. Radical tumorectomy could enhance the probability of survival of patients with thyroid carcinoma that had extensively invaded the larynx, esophagus and/or mediastinum. Postsurgical I(131) treatment and radiotherapy enhanced the probability of survival.

[Two cases of Castleman disease in the neck].

To investigate the clinical manifestation, clinicolpathological diagnosis and treatment of Castleman disease (CD) in the neck. The data of 2 patients with CD in the neck were reviewed retrospectively, together with literature summary, in order to analyze its clinical manifestation, pathological characteristics, clinical and pathological types, imaging characteristics and treatment. Two cases presented as solitary, painless neck masses and met the criteria of localized CD, and also had histological evidence of the hyaline vascular variant. Complete surgical resection was the first choice of treatment. One patient was alive without evidence of recurrence for ten years. The another patient had masses recurred in the neck one month after operation and followed to undergo curative chemotherapy, no recurrence in one-year follow-up. CD in the neck is a rare disease. The diagnosis for the disease is mainly based on pathological examination. Surgical resection is the main treatment, if necessary, need combination with radiotherapy and chemotherapy.

Whole-exome sequencing to decipher the genetic heterogeneity of hearing loss in a Chinese family with deaf by deaf mating.

Inherited deafness has been shown to have high genetic heterogeneity. For many decades, linkage analysis and candidate gene approaches have been the main tools to elucidate the genetics of hearing loss. However, this associated study design is costly, time-consuming, and unsuitable for small families. This is mainly due to the inadequate numbers of available affected individuals, locus heterogeneity, and assortative mating. Exome sequencing has now become technically feasible and a cost-effective method for detection of disease variants underlying Mendelian disorders due to the recent advances in next-generation sequencing (NGS) technologies. In the present study, we have combined both the Deafness Gene Mutation Detection Array and exome sequencing to identify deafness causative variants in a large Chinese composite family with deaf by deaf mating. The simultaneous screening of the 9 common deafness mutations using the allele-specific PCR based universal array, resulted in the identification of the 1555A>G in the mitochondrial DNA (mtDNA) 12S rRNA in affected individuals in one branch of the family. We then subjected the mutation-negative cases to exome sequencing and identified novel causative variants in the MYH14 and WFS1 genes. This report confirms the effective use of a NGS technique to detect pathogenic mutations in affected individuals who were not candidates for classical genetic studies.

Clinical manifestation and management of primary malignant tumors of the cervical trachea.

To explore clinical manifestation and therapies of primary malignant tumors of the cervical trachea, we retrospectively reviewed 31 patients with primary cervical tracheal malignant tumors diagnosed in the last 15 years by means of clinical manifestation, fiberoptic endoscopy, CT scanning and histopathological examinations. All of them were hospitalized and treated at the Second Xiangya Hospital, Central South University. Of them, 4 underwent emergent tracheotomy under local anesthesia, 9 were inserted with a laryngeal mask airway, 18 underwent tumorectomy under general anesthesia with endotracheal intubation, and of them 11 had tracheotomy during surgery. Of those 31 patients, tracheal malignant tumors in 9 cases were resected via laryngeal and retrograde tracheal incisions under endoscope; the tumors in 13 cases were excised via sleeve trachea resection and end-to-end anastomosis; those in 8 were removed by tracheofissure, and the tumor in 1 case was not excised surgically. Among the 30 resected patients, 20 patients received both radiotherapy and chemotherapy; 6 received radiotherapy only, and 4 did not receive any adjuvant therapies. During follow-up between 2 and 11 years, among 31 patients, there was no recurrence in 24 cases. Among the 7 deceased patients, 1 displayed multiple tracheal chondrosarcoma, 4 displayed adenoid cystic carcinoma, and 2 displayed squamous cell carcinoma. Emergency lower tracheotomy is necessary only when patients with tracheal, malignant tumors are in a critical condition. Sleeve trachea resection is the optimal therapy for tracheal malignant tumors. However, in the treatment of tracheal malignant tumors adjacent to the larynx or the involved trachea is over 6 cm in length, other surgeries shall be performed. Postoperative adjuvant radiotherapy and chemotherapy can achieve the same therapeutic effect as sleeve trachea resection.