Mutations in CLCN6 as a Novel Genetic Cause of Neuronal Ceroid Lipofuscinosis.

OBJECTIVE: The aim of this study was to explore the pathogenesis of CLCN6-related disease and to assess whether its Cl-/H+-exchange activity is crucial for the biological role of ClC-6. METHODS: We performed whole-exome sequencing on a girl with development delay, intractable epilepsy, behavioral abnormities, retinal dysfunction, progressive brain atrophy, suggestive of neuronal ceroid lipofuscinoses (NCLs). We generated and analyzed the first knock-in mouse model of a patient variant (p.E200A) and compared it with a Clcn6-/- mouse model. Additional functional tests were performed with heterologous expression of mutant ClC-6. RESULTS: We identified a de novo heterozygous p.E200A variant in the proband. Expression of disease-causing ClC-6E200A or ClC-6Y553C mutants blocked autophagic flux and activated transcription factors EB (TFEB) and E3 (TFE3), leading to autophagic vesicle and cholesterol accumulation. Such alterations were absent with a transport-deficient ClC-6E267A mutant. Clcn6E200A/+ mice developed severe neurodegeneration with typical features of NCLs. Mutant ClC-6E200A, but not loss of ClC-6 in Clcn6-/- mice, increased lysosomal biogenesis by suppressing mTORC1-TFEB signaling, blocked autophagic flux through impairing lysosomal function, and increased apoptosis. Carbohydrate and lipid deposits accumulated in Clcn6E200A/+ brain, while only lipid storage was found in Clcn6-/- brain. Lysosome dysfunction, autophagy defects, and gliosis were early pathogenic events preceding neuron loss. INTERPRETATION: CLCN6 is a novel genetic cause of NCLs, highlighting the importance of considering CLCN6 mutations in the diagnostic workup for molecularly undefined forms of NCLs. Uncoupling of Cl- transport from H+ countertransport in the E200A mutant has a dominant effect on the autophagic/lysosomal pathway. ANN NEUROL 2024.

Roles of KCNA2 in Neurological Diseases: from Physiology to Pathology.

Potassium voltage-gated channel subfamily a member 2 (Kv1.2, encoded by KCNA2) is highly expressed in the central and peripheral nervous systems. Based on the patch clamp studies, gain-of function (GOF), loss-of-function (LOF), and a mixed type (GOF/LOF) variants can cause different conditions/disorders. KCNA2-related neurological diseases include epilepsy, intellectual disability (ID), attention deficit/hyperactive disorder (ADHD), autism spectrum disorder (ASD), pain as well as autoimmune and movement disorders. Currently, the molecular mechanisms for the reported variants in causing diverse disorders are unknown. Consequently, this review brings up to date the related information regarding the structure and function of Kv1.2 channel, expression patterns, neuronal localizations, and tetramerization as well as important cell and animal models. In addition, it provides updates on human genetic variants, genotype-phenotype correlations especially highlighting the deep insight into clinical prognosis of KCNA2-related developmental and epileptic encephalopathy, mechanisms, and the potential treatment targets for all KCNA2-related neurological disorders.

Exploring the nurses' experiences in recognising and managing clinical deterioration in emergency patients: A qualitative study.

BACKGROUND: Emergency Department (ED) patients are particularly at a high risk of deterioration. The frontline nurses are key players in identifying and responding to deterioration events; however, few studies have sought to explore the whole process of recognition and management of clinical deterioration by emergency nurses. OBJECTIVES: The aim of this study was to explore the experiences of emergency nurses and provide a whole picture of how they recognise and manage clinical deterioration. METHODS: A qualitative descriptive study involving 11 senior nurses and seven junior nurses was conducted in the ED of a 3000-bed tertiary general hospital using semistructured interviews. The interviews were transcribed and thematically analysed. FINDINGS: Four salient themes emerged from the data analysis. The first, 'early recognition and response', revealed the importance of vital signs assessment in recognising and responding to clinical deterioration. The second, 'information transfer', depicted the skills and difficulties of transferring information in escalations of care. The third, 'abilities, education, and training', presented the abilities that emergency nurses should have and their perspectives on training. The fourth, 'support culture', described the major role of senior nurses in collaboration with colleagues in the ED. CONCLUSIONS: This study explored the experiences of emergency nurses in recognising and managing clinical deterioration. The findings illuminate the need to support the critical role of emergency nurses, with an emphasis on their abilities and continuous interprofessional collaboration training to improve the recognition and management of clinical deterioration.

Novel HCN1 Mutations Associated With Epilepsy and Impacts on Neuronal Excitability.

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channel plays a critical role in regulating the resting membrane potential and integrating synaptic transmission. Variants of HCN1 have been recognized as causes of epilepsy, and mutant HCN1 channels could act with loss-of-function (LOF), loss- and gain-of-function (LOF and GOF) and gain-of-function (GOF) mechanisms. However, phenotypes and pathogenesis of HCN1-related epilepsy are still poorly understood. This study enrolled five epileptic cases carrying five different HCN1 variants: two pathogenic variants (I380F and S710Rfs*71), two likely pathogenic variants (E240G and A395G), and a paternally inherited variant (V572A). Four variants were novel. Electrophysiological experiments revealed impaired biophysical properties of the identified mutants, including current densities and activation/deactivation kinetics. Moreover, three variants exerted effects on the biophysical properties of wild-type HCN1 channels in heterozygous conditions. Immunofluorescence experiments showed that two variants reduced the protein expression of HCN1channels in neurons. Neurons expressing E240G (GOF) variant showed increased input resistance. However, the variant of I380F (LOF) increased the neuronal firing rate, thus leading to neuronal hyperexcitability. In conclusion, the present study expands the genotypic and phenotypic spectrum of patients with HCN1-related epilepsy and clarifies the underlying mechanisms. We reported five new cases including four unreported likely/pathogenic variants. We provided assessments of biophysical function for each variant, which could help patients to receive individual therapy in the future. We confirmed that HCN1 variants contributed to neuronal hyperexcitability by regulating input resistance and the action potential firing rate, and we have shown that they can affect protein expression in neurons for the first time.

Clinical Characteristics, Classification, and Management of Adult Nasopharyngolaryngeal Hemangioma.

OBJECTIVES/HYPOTHESIS: To analyze the clinical features, classification, and treatment of adult nasopharyngolaryngeal hemangioma (ANPLH). STUDY DESIGN: Retrospective study. METHODS: From February 2009 to May 2020, 101 patients with ANPLH were reviewed and analyzed. RESULTS: Symptoms of ANPLH were frequently displayed as abnormal pharyngeal sensation and functional defection. According to lesion location, ANPLH was divided into five categories including nasopharyngeal, oropharyngeal, hypopharyngeal, laryngeal, and mixed types. The mixed type constitutes the highest portion, and the nasopharyngeal type is the least in our cohort. Most lesions could resect through natural cavity under endoscopy. Patients with mixed lesions had a higher rate of postoperative recurrence and planned multiple surgeries. Acceptable but not severe intraoperative and postoperative complications occurred in our patient cohort. CONCLUSIONS: Patients with ANPLH are always symptomatic and even functional defective, which can be classified into five categories based on lesion location. For these patients, endoscopic surgery through natural cavity is recommended to remove lesions with fewer complications and favorable clinical outcomes. LEVEL OF EVIDENCE: 4 Laryngoscope, 131:2724-2728, 2021.

Endoscopy-Assisted Transoral Approach to Resect Parapharyngeal Space Tumors: A Systematic Review and Meta-Analysis.

OBJECTIVES: By comparing the endoscopy-assisted transoral approach (EATA) with external approaches (EAs) for the resection of parapharyngeal space tumors (PPSTs), we determined whether the EATA has advantages in terms of operation time, intraoperative bleeding volume, postoperative hospitalization, drainage volume, and complications. At the same time, we summarized the surgical indications for the EATA. METHODS: Systematic literature retrieval was performed in the PubMed, Web of Science, Embase, CNKI, Wanfang, and CQVIP databases up to February 2020. We calculated the mean difference (MD) with a 95% confidence interval (CI) for continuous outcomes and pooled odds ratio (OR) with 95% CI for dichotomous outcomes. The measured outcomes were operative time, bleeding volume, postoperative hospitalization, drainage volume, and complications. RESULTS: Seven studies involving 318 patients were eligible. Of these patients, 145 patients underwent EATA and 173 patients underwent EA surgery. All the former tumors were benign and located medial or anteromedial to the carotid sheath except for the unrecorded tumors. Compared with EAs, the EATA significantly shortened the operation time (MD = -5.56 min, 95% CI: -9.58 to -1.55), shrank the bleeding volume (MD = -89.02 ml, 95% CI: -126.16 to -51.88), shortened the postoperative hospitalization (MD = -2.44 days, 95% CI: -3.37 to -1.51), reduced the drainage volume (MD = -32.97 ml, 95% CI: -36.24 to -29.70), and lowered the incidence of complications (OR = 0.30, 95% CI: 0.16 to 0.59). CONCLUSION: As for PPSTs, with an appropriate and precise patient selection, the EATA is a safe, effective, minimally invasive, and aesthetic surgical modality. Laryngoscope, 131:2246-2253, 2021.

Oxytocin mediates neuroprotection against hypoxic-ischemic injury in hippocampal CA1 neuron of neonatal rats.

Neonatal hypoxic-ischemic encephalopathy (NHIE) is one of the most prevalent causes of death during the perinatal period. The lack of exposure to oxytocin is associated with NHIE-mediated severe brain injury. However, the underlying mechanism is not fully understood. This study combined immunohistochemistry with electrophysiological recordings of hippocampal CA1 neurons to investigate the role of oxytocin in an in vitro model of hypoxic-ischemic (HI) injury (oxygen and glucose deprivation, OGD) in postnatal day 7-10 rats. Immunohistochemical analysis showed that oxytocin largely reduced the relative intensity of TOPRO-3 staining following OGD in the hippocampal CA1 region. Whole-cell patch-clamp recording revealed that the OGD-induced onset time of anoxic depolarization (AD) was significantly delayed by oxytocin. This protective effect of oxytocin was blocked by pretreatment with [d(CH2)51, Tyr (Me)2, Thr4, Orn8, des-Gly-NH29] vasotocin (dVOT, an oxytocin receptor antagonist) or bicuculline (a GABAA receptor antagonist). Interestingly, oxytocin enhanced inhibitory postsynaptic currents in CA1 pyramidal neurons, which were abolished by tetrodotoxin or dVOT. In contrast, oxytocin had no effect on excitatory postsynaptic currents but induced an inward current in 86% of the pyramidal neurons tested. Taken together, these results demonstrate that oxytocin receptor signaling plays a critical role in attenuating neonatal neural death by facilitating GABAergic transmission, which may help to regulate the excitatory-inhibitory balance in local neuronal networks in NHIE patients.

Home-based psychological nursing interventions for improvement of sleep quality and psychological health in patients with hypopharyngeal carcinoma undergoing surgical resections: a randomized trial.

BACKGROUND: The primary aim of this study was to determine the efficacy of a psychological nursing intervention for improving sleep quality and alleviating anxiety and depression among hypopharyngeal carcinoma patients undergoing surgical resections. METHODS: A total of 140 hypopharyngeal carcinoma patients undergoing surgical resections in our hospital from January 2017 to December 2020 were included and randomly assigned to a standard nursing group or psychological nursing group, with 70 cases per group. Sleep quality was assessed using the Pittsburgh sleep quality index (PSQI). The severity of anxiety symptoms and depressive disorders were evaluated using Zung's self-rating anxiety scale (SAS) and self-rating depression scale (SDS). The functional assessment of cancer therapy-general scale (FACT-G) tool was used to evaluate the quality of life. Fear of progression (FoP) was also scored. RESULTS: Participants with hypopharyngeal carcinoma in the psychological nursing group demonstrated greater improvements in the primary outcome of global sleep quality (P=0.002) at postintervention compared to the participants in the standard nursing group. The psychological nursing group participants showed remarkably greater improvements in the subscale scores of global sleep quality characteristics (secondary outcomes) including daytime dysfunction (P<0.001), subjective sleep quality (P=0.029), and sleep latency (P=0.006) at postintervention compared to those receiving standard nursing. Psychological nursing group participants had lower SAS and SDS scores compared to those in the standard nursing group (P<0.001). The results revealed that participants who received psychological nursing intervention exhibited higher scores of all FACT-G subscales and lower scores of FoP compared to participants receiving standard nursing intervention (both P<0.001). The global sleep quality scores shared positive associations with scores of SAS, SDS, and FoP (r=0.518, r=0.572, and r=0.395, respectively, P<0.001). CONCLUSIONS: These data provide evidence that home-based psychological nursing intervention could improve sleep quality and alleviate anxiety and depression among hypopharyngeal carcinoma patients undergoing surgical resections. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2100051463.

[Killing effect improved by fusion gene HRE1.Egr-1. yCDglyTK on gene-radio therapy of nasopharyngeal cancer in vitro].

OBJECTIVE: To construct hypoxia/radiation inducible promotor HRE1.Egr-1, and to observe its promotive effect on the expression of yCDglyTK gene in nasopharyngeal cancer HNE-1 cells and the anti-tumor effect of yCDglyTK and to lay an experimental foundation for further exploration of new gene-radio therapy of nasopharyngeal cancer. METHODS: pcDNA3.1(-)HRE1.Egr-1.yCDglyTK was constructed by gene recombination technique. Stable yCDglyTK-expressing HNE-1 cells were generated by transfecting the recombinant plasmid into the target cells with liposome. The expression of yCDglyTK was detected by Western blot in 4 groups: a normoxia group, a radiation group, a hypoxia group, and a hypoxia and radiation group. The killing effect of 5-FC in different circumstances was determined by MTT. RESULTS: The expression of yCDglyTK/5-FC gene in all the groups was significantly different(P<0.01),especially in the hypoxia and radiation group. The killing effect of 5-FC on HNE1 cells varied under different conditions, especially in the hypoxia and radiation group. CONCLUSION: Hypoxia and radiation can induce the activity of fusion promoter HRE1.Egr-1, and obviously promote the anti-tumor effect of yCDglyTK/5-FC system, suggesting that yCDglyTK may be a candidate suicide gene for gene-radio therapy of NPC.

[In vitro experimental study of killing Hep-2 cells of laryngeal cancer with suicide fusion gene CDglyTK].

OBJECTIVE: To study the killing effect of suicide gene CDglyTK combined with GCV or 5-FC on the human laryngeal carcinoma Hep-2 cell line in vitro. METHOD: Constructed plasmid pcDNA3.1 (-) CMV. CDglyTK was verified by enzyme digestion of Xho I /Hind III and automatic sequence analysis, then it was introduced into Hep-2 cells by electroporation to yield cells expressing CDglyTK stably after selecting with G418(400 ng/L) for 14 da. The expression of CDglyTK mRNA in transfected Hep-2 cells was tested by RT-PCR. Compared with Hep-2 cells transferred with pcDNA3.1(-), in vitro chemosensitivity of CDglyTK-expressing Hep-2 cells to 5-FC, GCV or 5-FC + GCV was detected by MTT assay. RESULT: The recombinant plasmid contained full-length coding region sequence of CD and TK gene. A anticipated 707 bp fragment was amplified from total RNA of CDglyTK-expressing Hep-2 cells by RT-PCR and a fusion protein of 59 000 was detected in cell extract from transfected Hep-2 cells. In vitro study growth of CDglyTK-positive Hep-2 cells were inhibited by 5-FC, GCV or 5-FC + GCV respectively, and the antitumour effect of 5-FC + GCV is superior to 5-FC or GCV. CONCLUSION: CDglyTK may be a candidate for treating human laryngeal cancer.