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 sinicize the English version of the rapid eye movement sleep behavior disorder symptom severity scale (RBDSSS) and to evaluate the reliability and validity of the Chinese version of RBDSSS (RBDSSS-C) among Chinese patients with rapid eye movement sleep behavior disorder (RBD).Methods:RBDSSS-C was ultimately formed through translation, back translation and revision according to the Brislin's translation model, including patient version (RBDSSS-PT) and bedpartner version (RBDSSS-BP). A questionnaire survey was conducted among 120 RBD patients to test the reliability and validity of the RBDSSS-C, using Cronbach’s α coefficient, Spearman-Brown coefficient, Spearman correlation analysis, content validity index and factor analysis. The correlation between RBDSSS-C and RBDQ-HK was examined.Results:For the Chinese version of RBDSSS-PT, the Cronbach’s α was 0.795, the split-half reliability was 0.756, and the test-retest reliability was 0.940. Item-level content validity indices (I-CVI) ranged from 0.833 to 1.000, and the scale-level CVI (S-CVI) was 0.937.For the Chinese version of RBDSSS-BP, the Cronbach’s α was 0.712, the split-half reliability was 0.813, and test-retest reliability was 0.950, with both I-CVI and S-CVI at 1.000.The scores of Chinese version of RBDSSS-PT and RBDSSS-BP were both significantly correlated with RBDQ-HK scores ( r=0.638, P<0.001 for RBDSSS-PT; r=0.639, P<0.001 for RBDSSS-BP). Factor analysis confirmed both single-factor structure for RBDSSS-PT and RBDSSS-BP.RBDSSS-PT showed χ2/ df=3.930, CFI=0.954, TLI=0.937, and RMSEA=0.093; RBDSSS-BP showed χ2/ df=8.300, CFI=0.975, TLI=0.966, and RMSEA=0.079. These results indicated adequate model fit. Conclusion:RBDSSS-C has good reliability and validity, and can be used as a reliable and effective tool to evaluate the severity of symptoms in Chinese RBD patients.

Objective:To sinicize the English version of the rapid eye movement sleep behavior disorder symptom severity scale (RBDSSS) and to evaluate the reliability and validity of the Chinese version of RBDSSS (RBDSSS-C) among Chinese patients with rapid eye movement sleep behavior disorder (RBD).Methods:RBDSSS-C was ultimately formed through translation, back translation and revision according to the Brislin's translation model, including patient version (RBDSSS-PT) and bedpartner version (RBDSSS-BP). A questionnaire survey was conducted among 120 RBD patients to test the reliability and validity of the RBDSSS-C, using Cronbach’s α coefficient, Spearman-Brown coefficient, Spearman correlation analysis, content validity index and factor analysis. The correlation between RBDSSS-C and RBDQ-HK was examined.Results:For the Chinese version of RBDSSS-PT, the Cronbach’s α was 0.795, the split-half reliability was 0.756, and the test-retest reliability was 0.940. Item-level content validity indices (I-CVI) ranged from 0.833 to 1.000, and the scale-level CVI (S-CVI) was 0.937.For the Chinese version of RBDSSS-BP, the Cronbach’s α was 0.712, the split-half reliability was 0.813, and test-retest reliability was 0.950, with both I-CVI and S-CVI at 1.000.The scores of Chinese version of RBDSSS-PT and RBDSSS-BP were both significantly correlated with RBDQ-HK scores ( r=0.638, P<0.001 for RBDSSS-PT; r=0.639, P<0.001 for RBDSSS-BP). Factor analysis confirmed both single-factor structure for RBDSSS-PT and RBDSSS-BP.RBDSSS-PT showed χ2/ df=3.930, CFI=0.954, TLI=0.937, and RMSEA=0.093; RBDSSS-BP showed χ2/ df=8.300, CFI=0.975, TLI=0.966, and RMSEA=0.079. These results indicated adequate model fit. Conclusion:RBDSSS-C has good reliability and validity, and can be used as a reliable and effective tool to evaluate the severity of symptoms in Chinese RBD patients.

Objective To investigate the shedding of CAV2-ΔE3-CGS after immunization and the background of canine adenovirus (CAV) infection, and to establish a dual fluorescent quantitative PCR detection method for rabies virus (RV) and canine adenovirus type 2 (CAV2). Methods A dual fluorescent quantitative PCR detection method was established by designing specific primers and probes for E1 gene of CAV and G gene of RV for the detection of CAV2-ΔE3-CGS. Oral swabs, anal swabs and environmental samples of stray dogs from experimental animal farm and dog detention center were tested. Results The standard curves generated by this method were Y=-3.351 × logX + 44.895, R² = .999 and Y=-3.413 × logX + 45.192, R²=0.996, respectively. The linear relationships were good, and the minimum detection limits were both 102 copies/μL. CAV2-ΔE3-CGS was not detected in experimental animal farm. CAV was detected in dog detention center, and the positive rates were 5.88% (5/85) in oral swabs, 8.24% (7/85) in anal swabs, and 4% (1/25) in environmental samples. Conclusion The dual fluorescent quantitative PCR method can be used for the detection of CAV2-ΔE3-CGS after immunization and the investigation of CAV infection. The present study has shown that no CAV2-ΔE3-CGS has been detected after immunization and CAV infection rate of stay dogs is low in Shanghai. CAV2-ΔE3-CGS oral immunization meets requirement and is applicable.

Restless legs syndrome is a common motor-sensory disease of the nervous system. The main manifestations are an urge to move the legs, usually accompanied by or thought to be caused by uncomfortable and unpleasant sensations in the legs. These symptoms may begin or worsen during periods of rest or inactivity such as lying down or sitting and be partially or totally relieved by movement, such as walking or stretching, at least as long as the activity continues. This article will systematically review the pathogenesis, clinical manifestations, diagnostic criteria, differential diagnosis and treatment of restless legs syndrome.

Parkinson's disease(PD)and restless legs syndrome(RLS)are relatively common movement disorders.There has been much debate over whether an etiological link exists between these two diseases and whether they share common pathophysiological mechanisms.PD and RLS may co-occur and respond well to dopaminergic agents, suggesting there is underlying dopamine dysfunction in both conditions.Despite the overlapping clinical features, the mechanisms underlying idiopathic RLS and RLS associated with PD may differ.In this article, we review studies related to the epidemiology, pathophysiology and genetics of PD with concurrent RLS, in order to provide evidence for exploring the link between RLS and PD.

Objective: To investigate the clinical outcomes of selective major aortopulmonary collaterals(MAPCAs) unifocalization and report histopathological findings in patients with pulmonary atresia with ventricular septal defect and major aortopulmonary collaterals(PA/VSD/ MAPCAs). Methods: The study enrolled 6 MAPCAs/VSD/PA patients with age ranged from 6 to 96 months and body weight ranged from 5.0 to 23.0 kg. These patients underwent selective MAPCAs unifocalization and primary repairs. Preoperative cardiac catheter, selective arteriography, cardiac CTA and intraoperative pathology were performed to identify different function, anatomic distribution and histopathology of MAPCAs. Results: 6 MAPCAs/VSD/PA patients underwent selective MAPCAs unifocalization and primary repair. No death occurred after operation and at follow-up which lasted for 1 to 20 months. Preoperative cardiac catheter, selective arteriography and intraoperative histopathology demonstrated distribution of functional MAPCAs similar to native pulmonary artery arborization and participating in arterial gas exchange. Functional MAPCAs were classified into elastic arteries according to histopathology. Conclusion There are two histological type of MAPCAs which play different roles. Selective unifocalization to functional MAPCAs which are classified into elastic arteries like native pulmonary artery is a safe and effective treatment approach for PA/VSD/MAPCAs.

Objective　To analyze the use of drugs andinfluencing factors of objective sleep disturbance among chronic insomnia patientsand to provide some normalization reference for clinical drug use. Methods　357 patients with definite chronic insomnia were selected. All patients were assessed with the Symptom Checklist （SCL-90），Hamilton Anxiety Scale （HAMA），Hamilton Depression Scale （HAMD），Pittsburgh sleep quality index （PSQI），Epworth sleepiness scale （ESS）. Polysomnography was used to evaluate objective sleep quality. Results　（1）The use of sedative and hypnotic drugs and other drugs that may affect sleep in the past 1 month before the first visit to the Sleep Center of our hospital was 53.50% for no drugs，46.5% for drugs and 18.21% for BzRAs among them. （2） The ESS scores of pharmacologic-therapy group was significantly lower than those in no-drugs group （Z=-2.463，P=0.014）. The total time sleep（Z=-3.521，P<0.001），Sleep efficiency（Z=-3.164，P=0.002），R%（Z=-3.044，P=0.003）were both significantly lower than those in no-drugs group. （3）The multivariate logistic regression results showed male （OR=1.938，95%CI 1.071～3.505，P=0.029），highly educated（OR=2.222，95%CI 1.196～4.130，P=0.012），anxiety state（OR=2.029，95%CI 1.018～4.043，P=0.044） and medication history（OR=1.974，95%CI 1.091～3.573，P=0.025） were the factors influencing SL>30 min. Age is the common factors for WASO>30 min（OR=1.052，95%CI 1.023～1.082，P<0.001），WASO>60 min（OR=1.068，95%CI 1.046～1.091，P<0.001） and SL>30 min and/or WASO>60 min （OR=1.068，95%CI 1.046～1.091，P<0.001）. Conclusion　Some patients have a high percentage of sleep-related medications before their first visit in our center，Most used was benzodiazepine receptor agonists. Gender，education，anxiety state and medication historywere the factors influencingsleep latency. Age is the common factors for difficulty maintaining sleep.

Objective To assess the epidemiology and clinical characteristics of sleep disorders in a single center in northwest China.Methods Using the International Classification of Sleep Disorders, 3rd Edition, all consecutive patients which were suspected as sleep disorders in Tangdu Hospital between January 2007 and December 2016, were included retrospectively.Results The average age of 8 037 patients was (46.59±15.83) years with male-female ratio 1∶1.24.Chronic insomnia was found in 3 848 (47.9%) patients, obstructive sleep apnea was found in 2 648 (32.9%) patients, narcolepsy was found in 294 (3.7%) patients, Kleine-Levin syndrome was found in 11 (0.1%) patients, circadian rhythm sleep-wake disorders were found in 14 (0.2%) patients, rapid eye movement behavior disorder was found in 193 (2.4%) patients, restless legs syndrome was found in 139 (1.7%) patients, periodic limb movement disorder was found in 109 (1.4%) patients, and other possible sleep disorders were found in 478 (5.9%) patients, respectively.Chronic insomnia and obstructive sleep apnea combided with somatic diseases.Conclusions Patients diagnosed by polysomnography in our single center suggested consultation rate of sleep disorders was increasing in past ten years, of which chronic insomnia and obstructive sleep apnea were dominant and comorbided with somatic diseases.