Objective Investigate the association between prefrontal cortex （PFC） function and hyperarousal in sleep deprivation. Methods EEG was used to investigate the association between PFC and hyperarousal in sleep deprivation. The data from the open-source database of A Resting-state EEG Dataset for Sleep Deprivation from Nanfang Hospital were analyzed， and the PSD analysis and the microstate analysis were used to compare brain functional activity during NS and sleep deprivation. Results The resting-state EEG data after awakening were collected from 37 participants， including 37 recordings after NS and 37 recordings after sleep deprivation for 24 hours. Compared with NS， there were increases in PSD values of all brain regions and PFC after sleep deprivation. Compared with NS， there were reductions in the duration and frequency of four microstates after sleep deprivation， while there were no significant changes in the coverage of the four states between NS and sleep deprivation. Conclusion This study shows that hyperarousal in PFC plays an important role in sleep deprivation， and good sleep can help to alleviate hyperarousal.

Objective Narcolepsy type 1 （NT1） is known to be associated with low levels of hypocretin-1 （Hcrt-1） in cerebrospinal fluid （CSF）. The standard method for Hcrt-1 measurement is radioimmunoassay （RIA） with imported reagents， but this antibody-dependent method is limited to radiation safety-certified lab， gradual radioactivity degradation， and slow turn-around time. The purpose of this study is to explore a non-radioactive， faster， and antibody independent domestic method in China for Hcrt-1 detection. Methods Repeated testing of cerebrospinal fluid from 14 clinically diagnosed NT1 patients and 10 non-narcolepsy patients was performed using liquid chromatography-tandem mass spectrometry （LC-MS/MS）technology，including the establishment and optimization of fundamental methodological procedures. The main steps involved the addition of non-radioactive isotope-labeled internal standards to the cerebrospinal fluid， followed by solid-phase extraction， mass spectrometry signal acquisition， and quantitative analysis. The results were then compared with the corresponding radioimmunoassay（RIA） findings. Results The LC-MS/MS method showed faster speed， and good linearity across a wider range of synthesized standard（5~2 500 pg/ml）， and good repeatability. Although this absolute-quantitation-based LC-MS/MS method and RIA method have different reading values in Hcrt-1 quantitation， they both can segregate NT1 group from non-NT1 group well. Conclusion Although larger cohorts are needed to set up a standard method in China，LC-MS/MS method is proved to be an easier， safer， faster， and possibly more accurate method for Hcrt-1 quantitation and detection for NT1 diagnosis.
Narcolepsy ; Radioimmunoassay

Objective:To study the distribution of species type, biotype and genotype of human Brucella isolated and identified in Nanjing. Methods:A total of 89 strains of human Brucella were collected from microbiology laboratories of three sentinel hospitals in Nanjing from 2017 to 2022. The species type was identified using biological methods and Brucella nucleic acid detection (BCSP31-PCR and AMOS-PCR). Further biotyping of Brucella melitensis isolates was conducted by serological results of A and M factors. Meanwhile, genotype analysis was performed using multiple-locus variable number tandem repeat analysis (MLVA), multilocus sequence typing (MLST) and single nucleotide polymorphism (SNP). Results:From 2017 to 2022, 89 strains of Brucella isolated and identified in Nanjing were all Brucella melitensis. Among them, Brucella melitensis biotype 3 accounted for 82.02% (73/89), and biotype 1 accounted for 17.98% (16/89). MLVA typing showed that 89 strains of Brucella melitensis belong to the "Eastern Mediterranean" cluster and could be divided into 50 MLVA genotypes; among which panel 1 had 3 genotypes, namely Type 42 (84.27%, 75/89), Type 63 (8.99%, 8/89) and Type 43 (6.74%, 6/89). The results of MLST-9 and MLST-21 were both ST8, and the core genome multilocus sequence typing (cgMLST) classified 89 strains into 11 genotypes. SNP analysis revealed a total of 4 013 SNP loci, with SNPs ranging from 0 to 409 across different strains, involving 59 SNP genotypes. Conclusions:The human Brucella strains isolated and identified in Nanjing are all Brucella melitensis, mainly biotype 3. The MLVA cluster is the "Eastern Mediterranean" cluster. The traditional MLST-9 and MLST-21 typing results are all ST8 type, while cgMLST divides all the strains into 11 genotypes with higher resolution.

Clinicians need to focus on various points in the diagnosis and treatment of insomnia.This article prescribed the treatment protocol based on the unique features,such as insomnia in the elderly,women experiencing specific physiologi-cal periods,children insomnia,insomnia in sleep-breathing disorder patients,insomnia in patients with chronic liver and kidney dysfunction.It pro-vides some reference for clinicians while they make decision on diagnosis,differentiation and treat-ment methods.

Objective To investigate the association between excessive daytime sleepiness (EDS) and cardiovascular health. Methods Data from the NHANES national representative database was used, and based on the EDS frequency questionnaire, the adults were divided into groups A，B，C, and D. The adults in group A had no daytime sleepiness (DS) group and chose the answer of "never", those in group B had mild DS and chose the answer of "rarely, sometimes (1-4 times a month)", those in group C had EDS and chose the answer of "often (5-15 times a month)", and those in group D had high-frequency EDS and chose the answer of "almost always (16-30 times a month)". The logistic regression analysis was used to estimate the degree of association of EDS with cardiovascular risk factors (hypertension, diabetes, hypercholesterolemia, and obesity) and cardiovascular event outcomes (including coronary artery disease, congestive heart failure, and stroke). Result This study included 12 786 unweighted participants from the four NHANES study periods of 2005-2008 and 2015-2018. Compared with the adults without DS, the adults with high-frequency EDS had a significantly higher proportion of the adults with a young age, female sex, a high body mass index, abdominal obesity, smoking, drinking, obstructive sleep apnea, sleep duration<6 hours,existing sleep problems, or a comorbidity of depression. The logistic regression analysis showed that high-frequency EDS was an independent risk factor for coronary artery disease (OR=1.02,95% CI 1.00-1.04,P=0.044) and stroke (OR=1.02, 95%CI 1.00-1.03,P=0.031). Conclusion High-frequency EDS is an independent risk factor for coronary artery disease and stroke. EDS should be taken seriously in clinical practice, and cardiovascular health management should be strengthened for EDS patients.

Restless legs syndrome （RLS） is a common sensory-motor disorder of the nervous system， and its mechanism remains unknown， which may be related to neuropsychiatric diseases， nutritional and metabolic diseases， kidney diseases， and pregnancy. We report a special case of RLS which was diagnosed prior to hyperthyroidism and experienced worsening of the RLS symptoms. We aim to discuss the mechanism of hyperthyroidism aggravating RLS， helping clinicians have a better understanding of the influencing factors for RLS.
Hyperthyroidism ; Comorbidity

ObjectiveTo investigate the effect of Bufeitang on intestinal flora of rats with lung Qi-deficiency syndrome of chronic obstructive pulmonary disease（COPD）， and to explore the mechanism of traditional Chinese medicine in regulating intestinal flora and thus restoring the balance of lung-gut axis. MethodA total of 84 rats were randomly divided into 7 groups， including blank group， model group， fecal bacterial transplantation（FMT） group， dexamethasone group and low， medium and high dose groups of Bufeitang， 12 rats in each group. Except for the blank group， cigarette and sawdust fumigation combined with intratracheal instillation of lipopolysaccharide（LPS） were used to establish the COPD rat model with lung Qi-deficiency syndrome in all other groups. The low， medium and high dose groups of Bufeitang were intragastric administrated with Bufeitang（3.645， 7.29， 14.58 g·kg^-1）， the FMT group was given fecal bacteria liquid enema（10 mL·kg^-1）， dexamethasone group was given dexamethasone acetate tablet suspension by gavage（0.135 mg·kg^-1）， the blank group and model group were given equal amount of distilled water. Fresh feces were collected after 28 d of continuous intervention for 16S rRNA gene sequencing. Lung and colon tissues were stained with hematoxylin-eosin（HE） for pathomorphological observation， and enzyme-linked immunosorbent assay （ELISA） was performed to detect the contents of tumor necrosis factor-α（TNF-α） and interleukin-8（IL-8） in lung tissues. ResultCompared with the blank group， the model group showed severe abnormal lung tissue structure with alveolar atrophy and collapse accompanied by severe inflammatory cell infiltration. Compared with the model group， the extent of injury was significantly improved， and inflammatory cell infiltration was reduced with basically normal alveolar structure in the high dose group of Bufeitang. Compared with the blank group， the model group had severely abnormal colonic tissue structure， the epithelial cells in the mucosal layer were eroded and shed， the number of inflammatory cells increased， the submucosal layer was edematous and the gap was enlarged. Compared with the model group， the extent of damage was significantly improved in the medium and high dose groups of Bufeitang， the epithelial cells in the mucosal layer were neatly and closely arranged， with only a small amount of inflammatory cell infiltration and no significant degeneration. Compared with the blank group， the TNF-α and IL-8 levels of lung tissue in the model group were significantly increased（P<0.01）. Compared with the model group， the TNF-α and IL-8 levels of lung tissues in the low， medium and high dose groups of Bufeitang were significantly decreased（P<0.01）. Bufeitang significantly modulated the number of bacteria species as well as alpha and beta diversity of model rats， corrected the return of intestinal flora to normal abundance and diversity， and positively regulated 4 differential phyla（such as Firmicutes， Proteobacteria） and 13 differential genera（such as Turicibacter， Lactobacillus， Anaerobiospirillum， Intestinimonas） in COPD model rats with lung Qi-deficiency syndrome， and down-regulated 2 carbohydrate metabolic pathway functions， including the pentose phosphate pathway（non-oxidative branch） Ⅰ and the Calvin-Benson-Bassham cycle. ConclusionBufeitang can modulate the abundance and diversity of intestinal flora species， affect the function of metabolic pathways， repair the structure of lung and colon tissues， regulate the level of inflammatory factors， and thus improve COPD with lung Qi-deficiency syndrome. The mechanism may be related to its regulation of inflammation-related intestinal flora to restore the balance of lung-gut axis in COPD with lung Qi-deficiency syndrome.

The treatment of insomnia currently involves drug and non-drug therapies, with varying treatment outcomes. There is no consensus on the treatment option for insomnia in patients in different post-stroke phases. This article provides a detailed literature review of common insomnia treatment methods based on their application in insomnia in the acute phase, subacute phase, and recovery phase following stroke, aiming to provide guidance for the treatment of post-stroke insomnia. Currently, there is a lack of high-quality randomized controlled trials on the treatment of stroke-related insomnia. Existing research does not support the use of benzodiazepine drugs for the treatment of stroke-related insomnia. Studies suggest that sleep hygiene education and cognitive behavioral therapy for insomnia, non-benzodiazepine drugs, melatonin receptor agonists, antihistamine H1 receptor medications, certain sedative antidepressants, orexin receptor antagonists, and traditional Chinese medicine therapy can improve sleep quality in the subacute and recovery phases after stroke, facilitating the recovery of neurological function in patients. However, there is insufficient evaluation of the impact on neurological function recovery and the side effects.

Objective To analyze the electrophysiological characteristics of patients with severe Guillain-Barré syndrome and the related factors causing poor prognosis,and evaluate the value of neuroelectrophysiology in predicting poor prognosis. Methods The clinical data of 78 GBS patients admitted to our hospital from March 2020 to May 2022 were selected,divided into mild (n=52) and severe (n=26) according to the severity of the disease,and the good group (n=45) and bad group (n=33) according to the prognosis;Compare the clinical data of different groups,compare the neuroelectrophysiological characteristics of patients in different disease groups,analyze the relevant factors causing poor prognosis,build a line chart prediction model and verify it;analyze the value of electrophysiological characteristics for poor prognosis through the subject's working characteristics (ROC) curve. Results A comparison of the clinical data of two groups of patients with different severity found that the incidence of facial paralysis,chest tightness,diarrhea,mechanical ventilation,autonomic nervous dysfunction and other autoimmune diseases in the severe group was significantly higher than that of patients in the mild group (P<0.05). By comparing the electrophysiological characteristics of the two groups,it was found that the total peroneal nerve conduction rate and tibia motor conduction rate of the severe severe group were significantly lower than that of the mild group,while the F-wave abnormality rate and dCMAP wave amplitude abnormality rate were significantly higher than that of the mild group (P<0.05);single-factor and multi-factor results showed that the pre-hospital course of hospitalization was >2 weeks,elevated cerebrospinal fluid protein,and MRC score ≤30 at hospital,mechanical ventilation,combined with other autoimmune diseases,slowing of the total peroneal nerve conduction rate,slowing down the tibial motion conduction rate,increased F-wave abnormality rate,increased amplitude abnormality of dCMAP and axial cord injury are all risk factors affecting poor prognosis (OR values are greater than 1,P<0.05);the total score of the line chart prediction model is 613 points,and the corresponding adverse prognosis probability is 74.56%,which has good distinction,accuracy and effectiveness;the ROC curve obtains the total peroneal nerve conduction rate,tibial nerve motion conduction rate,F-wave abnormality,dCMAP wave amplitude abnormality four the area,positive/negative prediction value,sensitivity and specificity under the ROC curve of the joint prediction are significantly higher than that of individual predictions,with better prediction performance. Conclusion Analyzing the electrophysiological characteristics of severe GBS patients is conducive to a clearer understanding of the severity of patients' condition. It has a very important predictive value for poor prognosis,and is worth popularizing and applying in clinic.

Objective Patients with obstructive sleep apnea (OSA) can present with excessive daytime sleepiness (EDS),insomnia,and other symptom subtypes. It remains unclear whether proteomics differs in patients with OSA with EDS and insomnia. Therefore,we examined the serum proteomics of patients with OSA with different clinical subtypes,in order to identify potential biomarkers for the classification of clinical subtypes of OSA,and provide insights into the molecular biological mechanisms underlying different subtypes of the disease. Methods We included six patients with OSA (three with EDS and three with insomnia) at the Second Affiliated Hospital of Air Force Medical University. All the patients underwent overnight polysomnography (PSG) and sleep scale evaluation,followed by blood sample collection on the next morning after PSG. Differentially expressed proteins were selected by label-free quantification,and bioinformatics analysis was performed. Results We identified a subset of 34 proteins that were differentially expressed between OSA with EDS and with insomnia. Compared with the OSA with insomnia group,the OSA with EDS group showed 20 up-regulated and 14 down-regulated serum proteins. The bioinformatics analysis revealed that the main functions of these proteins focused on DNA damage,reactive oxygen species elimination,immune regulation,and inflammatory response,and they were mainly involved in intracellular transport,secretion,and vesicle transport and other biological processes. The differentially expressed proteins were mainly localized in the cytoplasm,which might be secretory proteins. The APRT-AK1 interaction pathway might play an important role in different clinical subtypes of OSA. Conclusion We determined 34 differentially expressed proteins between OSA with EDS and OSA with insomnia,such as significantly up-regulated DDI2 and PRDX6 in the serum of patients with OSA with EDS,which can identify different clinical subtypes of OSA. In addition,the APRT-AK1 interaction pathway might play an important role in different subtypes of OSA.