OBJECTIVES: The purinergic receptor P2X2 (P2RX2) encodes an ATP-gated ion channel permeable to Na⁺, K⁺, and especially Ca²⁺. Loss-of-function mutations in P2RX2 are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a p2rx2 knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of p2rx2 deficiency on the auditory system, providing a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies. METHODS: Two CRISPR targets (sgRNA1 and sgRNA2) spaced 47 bp apart were designed within the zebrafish p2rx2 gene. Synthesized sgRNAs and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group. RESULTS: A stable p2rx2 knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all P<0.001), indicating reduced hearing sensitivity. CONCLUSIONS We successfully generated a p2rx2 loss-of-function zebrafish model using CRISPR/Cas9 technology. p2rx2 deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotype's restriction to the otolith region suggests tissue-specific roles of p2rx2 in sensory organs. This model provides a valuable tool for elucidating the molecular mechanisms of P2RX2-related hearing loss and for screening otoprotective drugs and developing gene therapies.
Animals ; Zebrafish/genetics* ; Receptors, Purinergic P2X2/deficiency* ; CRISPR-Cas Systems/genetics* ; Gene Knockout Techniques ; Phenotype ; Zebrafish Proteins/genetics* ; Disease Models, Animal

AIM: To explore the current status, research hotspots, and trends of global uveitis research to provide a theoretical basis and references for researchers in the field of uveitis, and promote further development in this area.METHODS: Relevant literatures on uveitis were retrieved from the China National Knowledge Infrastructure(CNKI)database, Wanfang database, and Web of Science core collection database since their establishment until 24 August 2023. The country/publishing institutions, research authors, high-frequency keywords, and burst keywords were visual analyzed by using software such as GraphPad Prism 9, CiteSpace 6.2. R2, and VOSviewer.RESULTS: Research teams for uveitis have been formed in various countries globally. The top three countries in terms of publications are the United States of America(7 585 papers), the United Kingdom(2 412 papers)and Germany(1 679 papers). The top three foreign institutions in terms of publications are Harvard University, Oregon Health & Science University, and Moorfields Eye Hospital, while the top three domestic institutions are Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine, Chongqing Medical University, and Zhongshan Ophthalmic Center, Sun Yat-sen University. The analysis of high-frequency keywords and burst keywords in Chinese and English shows that research hotspots mainly focus on exploring pathogenesis and different treatment methods for uveitis. The research hotspots related to uveitis treatment are transitioning to molecular biology-related research topics, such as molecular biological signaling pathways(NF-κB signaling pathway with a strength value of 22.89), biological agents(adalimumab with a strength value of 32.21), and tumor necrosis factor(with a strength value of 48.44). Related research is also expanding to basic experiments on relevant rats.CONCLUSIONS: In recent years, the research hotspots and trends of global uveitis mainly focus on precise diagnosis, pathogenesis, and more effective treatment methods. It is important for more scholars to dedicate themselves to uveitis-related research in the future to make breakthroughs and progress in the field. More large-scale and multicenter clinical studies on uveitis can provide high-quality research evidence.

OBJECTIVE：To reappraise systematic review/Meta-analysis （SRs/MAs）of the efficacy and safety of glucagon-like peptide 1（GLP-1）receptor agonist in the treatment of type 2 diabetes mellitus （T2DM），and to provide evidence-based reference for clinical use of these drugs in the treatment of T 2DM. METHODS ：Retireved from Cochrane library ，PubMed，Embase，CBM， Wanfang database and CNKI ，systematic review/Me ta-analysis about GLP- 1 receptor agonist in the treatment of T 2DM were collected during the inception to Dec. 2019. After data extraction of literatures met inclusion and exclusion criteria ，GRADE system was used to evaluate the quality of evidence included in the study ，and the evidence of efficacy and safety outcome indexes were summarized. RESULTS ：Finally 31 literatures were included ，involving 91 outcome indexes ，and GRADE evidence quality was medium，among which 4（4.4％）were very-low-quality ，33（36.3％）were low-quality ，45（49.5％）were medium-quality ，and 9 （9.9％）were high-quality outcome indicators. The results of evidence summary showed that GLP- 1 receptor agonists were better than or similar to placebo and other oral hypoglycemic drugs ， better than dipeptidyl peptidase 4 （DPP-4） inhibitors in . reducing the level of HbA 1c；better than or similar to placebo ， JDZX2015240） better than other oral hypoglycemic agents and DPP- 4 2276299207@qq.com inhibitors in reducing the level of fasting glucose ；similar to DDP-4 inhibitors，higher than or similar to placebo ，lower than other oral hypoglycemic dru gs in the incidence of hypoglycemia；higher than other oral hypoglycemic drugs ，placebo and DPP- 4 inhibitors in the incidence of diarrhea and nausea ； higher than other oral hypoglycemic drugs and placebo in the incidence of vomiting. CONCLUSIONS ：The evidence quality of systematic review/Meta-analysis about GLP- 1 receptor agonist in the treatment of T 2DM are moderate. These drugs have good clinical efficacy in the treatment of T 2DM，but their safety are not as good as placebo or other oral hypoglycemic drugs.

Objective To investigate the affecting factors on auditory and speech performances in preschool children with unilateral cochlear implantation (CI) .Methods The clinical data of the preschool children (n=165) with unilateral cochlear implantation in the Second Xiangya hospital from January 2006 to April 2013 were collected . These children received rehabilitation according to the method recommended by the China Rehabilitation Research Center for Deaf Children ,and the data were analyzed retrospectively .The categories of auditory performance (CAP) and speech intelligibility rating (SIR) were used to assess their auditory and speech performances .The relationships between the performance and gender ,implanted age ,genotype ,inner ear malformation ,history of hearing aid were evaluated .Results Implanted ages and genotypes were associated with the auditory and speech performance of par‐ticipants (P<0 .05) ,while genders ,hearing aid experience ,and inner ear malformations(enlarged vestibular aque‐duct syndrome ,EVAS)were not significant related (P<0 .05) .Children were found to have achieved better CAP and SIR growths when CI was implanted during 1~3 years old and 2~4 years old ,respectively (P<0 .05) .The outcomes of CI recipients with GJB2 mutation were significantly better than those of the GJB2-nonrelated CI recipi‐ents (P<0 .05) .Conclusion This study provides evidence that CIs during first 1~3 years old having better auditory rehabilitation results than those of during 4~6 years old ,and CIs during 2~4 years old obtaining a better speech development in the first 12 months after operation .Deaf children with GJB2 mutation show better auditory and speech performances after CIs than those of the peers without GJB2 mutation .CIs can be effectively performed in deaf children associated with EVAs as in those without EVAS .

OBJECTIVE: To explore the clinical characteristics and therapies for esophageal perforation complicated with lethal massive hemorrhage caused by esophageal foreign body. METHOD: To retrospective analysis the treatment of massive hemorrhage at the carotid artery or aorta caused by esophageal foreign body in forty seven patients, Foreign body characters, surgical approaches, and postsurgical management were summarized. RESULT: Among 24 patients with cervical esophageal foreign body, the object was removed either by esophagoscopy or through lateral cervical incision. After controlling carotid artery hemorrhage and repairing Fistula of artery from cervical incision, 19 patients survived. For the 23 patients with thoracic esophageal foreign body accompanied with aorta hemorrhea, thoracotomy was performed to remove the foreign body and repair the aortic fistula. Only 3 of these 23 patients recovered from the emergent surgery, other 20 patients died. CONCLUSION For the patients with esophageal foreign body inducing large vessel impingement, the most reliable therapeutic method is surgical repairing of arterial perforation and extraction of the foreign body via cervical or thoracic incision. Carotid ligation should be considered in patients with recurrent carotid hemorrhage. For the patient with mediastinitis, esophageal exclusion is recommended to prevent infection and to promote healing of aortic perforation after aortic fistula repairing.
Adult ; Esophageal Perforation ; etiology ; surgery ; Esophagus ; Female ; Follow-Up Studies ; Foreign Bodies ; complications ; Hemorrhage ; etiology ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVE: To screen and identify the frequency and characteristic of mutation in stereocilium-related gene Taperin of Chinese prelingual nonsyndromic hearing impairment with DNA microarray combined with PCR. METHOD: One hundred and thirty-four patients of prelingual nonsyndromic deafness and one hundred health individuals in China were investigated in this study. Genomic DNA was extracted from the patients and was subjected to DNA microarray to screen mutations in 4 most common genes. The samples that carried none of the common mutant alleles were subjected to PCR and sequenced to detect mutations in Taperin gene. RESULT: Ninteen out of one hundred and thirty-four patients of prelingual nonsyndromic deafness were detected carring common deafness gene with DNA microarray. Taperin gene were detected in one hundred and fifteen patients with PCR. A187S was detected in Taperin as hetrozygous state in 2 patients and their unaffected members of their family. It occurred at the evolutionary conservation of the amino acids of taperin according to alignment analysis. Two polymorphism, 157C>T and 318C>T, were found in the patients and the control group. CONCLUSION A novel Taperin mutation, A187S was detected in Chinese patients with prelingual nonsyndromic hearing loss, which may be relevant to hearing loss. Two polymorphism, 157C>T and 318C>T, were found in Chinese in our research. The carrier frequency for Taperin mutation is about 1.74% of prelingual nonsyndromic deafness in Chinese patients.
Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; China ; Deafness ; genetics ; Female ; Humans ; Male ; Mutation ; Proteins ; genetics ; Young Adult

OBJECTIVE: To study the correlation of CD44 with epithelial-mesenchymal transition(EMT) and metastasis in nasopharyngeal cancer cells, and explore the possible mechanism of CD44 regulates EMT and metastasis in nasopharyngeal cancer cells. METHOD: The CD44 and EMT-associated proteins in 5-8F and 6-10B nasopharyngeal cancer cell lines were assayed by Western blotting. The erasion trace test was performed to observe the migratory ability of 5-8F and 6-10B nasopharyngeal cancer cells. Using lipid-mediated DNA transfection technique, the low metastatic nasopharyngeal cancer cells 6-10B were transfected in vitro with plasmid which contained CD44 gene, and then new nasopharyngeal cancer cells were obtained. The CD44 and EMT-associated proteins in 6-10B, empty vector transfected and CD44-transfected cells were assayed by Western blotting. The erasion trace test was performed to observe the alteration of migratory ability of nasopharyngeal cancer cells before and after CD44 transfection. RESULT: The expression of CD44 and EMT-associated protein MMP-9 in 5-8F was higher than that in 6-10B, but EMT-associated protein E-Cadherin in 5-8F was lower than that in 6-10B. The migratory ability of 5-8F was higher than that of 6-10B. The expression of CD44 and MMP-9 were significantly higher in the CD44-transfected nasopharyngeal cancer cells than in the control groups. Compared with control groups, the migratory ability of CD44-transfected nasopharyngeal cancer cells was significantly increased. CONCLUSION CD44 positively regulates the metastatic ability of nasopharyngeal cancer cells, which is relevant to the process of EMT.
Carcinoma ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Humans ; Hyaluronan Receptors ; genetics ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Transfection

OBJECTIVE: To investigate the clinical manifestation, treatment and prognosis of extramedullary plasmacytoma(EMP) in the upper airway, and to improve the diagnosis and outcome of EMP treatment. METHOD: Clinical data of 26 EMP cases were reviewed retrospectively, and then compared with multiple myeloma(MM) patients presenting with lesions in upper airway. RESULT: Of 26 cases, 9 cases with the tumors occurred in nasal cavities, 7 in nasal sinuses, 6 in pharyngeal, 4 in throat, mainly manifesting with local masses and relevant symptoms. The manifestations of clinical, endoscopy findings and pathologic results in EMP patients were not distinguishable from the lesions of MM patients, while MM patients often accompanied by other findings, such as anemia and bone damage. Involvement of neck lymph nodes was more common in MM patients than in EMP patients. Ten patients were treated with surgery, and 16 patients with surgery and radiotherapy. Of the seven EMP patients with involvement of neck lymph nodes, four patients received additional chemotherapy besides surgery and radiotherapy, and no local relapse and MM happened in them, while of the three patients only received surgery and radiotherapy, one local relapse were found and one progressed to MM. CONCLUSION The diagnosis of EMPs mainly depends on pathological results. The judgment of pathologists and application of molecular biology technology are vital for the diagnosis of EMP in upper airway, and MM must be excluded very carefully in the diagnosis of EMP. Surgery combined with radiotherapy is the main treatment for EMP in the upper airway, and the prognosis is good but the follow-up should be taken. Besides surgery and radiotherapy, chemotherapy is beneficial for the EMP patients accompanied with lesions in neck lymph nodes.
Adult ; Aged ; Female ; Humans ; Lymph Nodes ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; pathology ; therapy ; Nasal Cavity ; pathology ; Plasmacytoma ; diagnosis ; pathology ; therapy ; Prognosis ; Respiratory Tract Neoplasms ; diagnosis ; pathology ; therapy ; Retrospective Studies ; Young Adult

OBJECTIVE: To establish the kanamycin-induced deafness model in SD rats, and to investigate the expression and significance of transmembrane protease, serine 3 (TMPRSS3) in the cochlea following kanamycin ototoxicity. METHODS: A total of 40 male SD rats were randomly divided into 4 groups. The experimental rats received intramuscular kanamycin sulfate for 3, 7, and 14 consecutive days, and the control group were treated with normal saline for 14 days. Auditory brainstem responses (ABR) were obtained before and after the kanamycin administration. The expression of TMPRSS3 in the cochlea was identified and detected by immunohistochemistry and Western blot. RESULTS: Kanamycin-induced deafness model in the SD rats was successfully established. ABR thresholds were increased and the expression of TMPRSS3 in the cochlea was reduced after the kanamycin injection (P<0.01). CONCLUSION TMPRSS3 may play an important role in normal cochlea function and involve in the process of aminoglycoside antibiotics induced deafness.
Animals ; Anti-Bacterial Agents ; toxicity ; Cochlea ; drug effects ; metabolism ; Deafness ; chemically induced ; metabolism ; Disease Models, Animal ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Kanamycin ; toxicity ; Male ; Membrane Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serine Endopeptidases ; metabolism

OBJECTIVE: To observe the expression of proteinase transmembrane protease, serine 3 (TMPRSS3) in mouse cochlea, and to investigate the significance of TMPRSS3 in the inner ear. METHODS: The protein expression of TMPRSS3 in C57/BL mouse cochlea was identified and detected by immunohistochemistry and immunofluorescence. Different cochlear tissues, such as spiral ganglion neurons, corti organ, stria vascularis and so on, were separated to detect the gene expression of TMPRSS3 by real-time fluorescence quantitative polymerase chain reaction (qPCR). The cochlear tissues with different ages were collected and the expression of TMPRSS3 mRNA was detected by qPCR. RESULTS: TMPRSS3 was mainly expressed in the spiral ganglion neurons, and there was TMPRSS3 mRNA in the cochlea in groups with different age. The expression level of TMPRSS3 mRNA was much weaker. CONCLUSION The distribution of TMPRSS3 was observed in many regions of the mouse cochlea, but mainly in the spiral ganglion neurons. This indicates that TMPRSS3 may be involved in the physiological functional regulation of the spiral ganglion neurons.
Animals ; Cochlea ; metabolism ; Female ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mice, Inbred C57BL ; RNA, Messenger ; genetics ; metabolism ; Serine Proteases ; genetics ; metabolism