Objective To construct the eukaryotic expression vectors of fibroblast growth factor receptor 3(FGFR3) MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN, and to detect their expressions in human chronic myeloid leukemia(CML)K562 cell line.Methods The full-length FGFR3 (fgfr3-WT)and dominant negative FGFR3 (fgfr3-DN)were amplified by polymerase chain reaction (PCR). The two genes were respectively digested with EcoRⅠand BamHⅠ,and then ligated into MSCV/puro to construct the recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN which were tranduced into K562 cells by LipofectaminTM 2000 after PCR,double digestion and DNA sequencing.The expressions of FGFR3 protein in K562 cells were detected by Western blotting and flow cytometry. Results The recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN were amplified by PCR method, and the results showed fgfr3-WT of 2 400 bp and fgfr3-DN of 1 200 bp had been successfully cloned into MSCV-puro vector. The 2 400 bp fragment was oblained after double digestion of recombinant plasmid.The sequencing results showed that the size of fgfr3-WT was 2 400 bp which was the same as the sequence from GeneBank.Fgfr3-DN of 1 200 bp was also in conformity with the expected sequence.Compared with control (K562 MSCV)group,the expression level of FGFR3-WT in MSCV/puro-fgfr3-WT transfection (K562-WT)group was increased to above 10 times.There was high expression of FGFR3-DN in MSCV/puro-FGFR3-DN transfection (K562-DN)group,but there was no expressions in control(K562 MSCV)group and K562-WT group.The flow cytometry results showed that the high expressions of FGFR3-WT were in 57.5% cells in K562-DN group and the high expressions of FGFR3-DN were in 41.5% cells in K562-DN group. Conclusion The K562 cell lines highly expressing FGFR3-WT and FGFR3-DN are constructed successfully.

Objective To identify and rename one strain stored in National Center for Medicine Culture Collections ( CMCC) and used for tracheitis vaccine production. Methods The test strain CMCC (B)29108 and the type strain DSM30007T were cultured on NA medium. Characteristics in morphology, physiology, biochemistry and fatty acid profile were compared between the two strains. Phylogenetic analysis was based on 16S rRNA and rpoB gene sequences, together with the DNA-DNA hybridization assay. Results A Comparative analysis of a partial sequence of the 16S rRNA gene sequence revealed that the CMCC( B) 29108 strain was closed to the Acinetobacter species and showed the highest similarity with the type strain Acinetobacter baumannii DSM30007T. Moreover, the CMCC(B)29108 strain was highly similar to type strain DSM30007T in morphology, physiology, biochemistry and fatty acid profile. On the phylogenetic tree based on 16S rRNA and rpoB gene of all Acinetobacter members, the CMCC(B)29108 strain steadily clustered into one independent branch only with the DSM30007 T strain with a DNA-DNA hybridization value of 100%. Conclusion The CMCC(B)29108 strain that is one of the strains used for the production of tracheitis vac-cine should be assigned to the species of Acinetobacter baumannii based on its phenotypic, phylogenetic and chemotaxonomic characteristics.

Objective: To survey the prevalence and distribution of sleep disorders in patients with Parkinson disease (PD) and to analyze the influencing factors. Methods: The prevalence and distribution of sleep disorders were surveyed with Parkinson Disease Sleep Scale (PDSS) among 206 PD patients. The association of sleep disorders with age, course of disease, cognitive function, motor function, depression, and the equivalent dose of levodopa (LED) was analyzed. Results: The overall PDSS score in 206 patients was (116.9±21.4). The three most frequent items of sleep disorders were the overall sleep quality(181/206, 87.9%), difficulty in maintaining sleep(160/206, 77.7%)and nocturnal enuresis(151/206, 73.3%); the three least frequent items were early awaking(87/206, 42.2%), urinary incontinence(56/206, 27.2%)and hallucination(44/206, 21.4%). The three items with the lowest average scores were nocturnal enuresis(6.9±3.1), difficulty in maintaining of sleep(7.1±2.7)and overall sleep quality(7.1±2.0); three items with the highest average scores were audiovisual illusion(9.3±1.8), incontinence caused by motion disability(9.0±2.1) and early awaking with upper and lower limb pain(8.7±2.1). PD patients were divided into group 1 [Hoehn-Yahr(H&Y) stage 1.0-1.5], group 2 (H&Y stage 2.0-2.5) and group 3 (H&Y stage 3.0-4.0). One-way analysis of variance or non-parametric test showed that there were significant differences in the course of disease(F=21.91, P=0.00), total score and the subscale scores of Unified Parkinson Disease Rating Scale(UPDRS, UPDRS Ⅰ-Ⅳ) (F=90.67, χ²=12.86, F=31.20, F=60.17, χ²=24.01, all P<0.01), the Hamilton Rating Scale for Depression(HAMD) scores (χ²=11.06, P=0.00), LED (F=14.93, P=0.00) and Minimum Mental State Examination(MMSE) scores (χ²=9.81, P=0.01) among three groups. There was no significant difference in age and PDSS scores among three groups. The results showed that there were significant differences in terms of restless state and nocturnal dysphoria (F=5.12, P=0.01; F=3.27, P=0.04) between group 1 and group 3. The linear regression analysis showed that the HAMD and the LED scores had the greatest influence on PDSS score (R²=0.142, 0.196). Conclusion PD patients have a variety of sleep symptoms. The treatment of large doses of dopamine and depression contribute to the occurrence of PD sleep disorders.

Objective:To investigate the characteristics of white matter lesions(WML)found by magnetic resonance imaging(MRI)and the relationship with clinical features in patients with Parkinson's disease(PD).Methods:This was a retrospective study by using a method of MRI T2WI-FLAIR.The WML in 87 PD patients were evaluated by using the Fazekas scale and Scheltens scale.Patients were divided into the early PD group[n=47, Hoehn-Yahr(H-Y)stage 1.0-2.0] vs.the middle-advanced PD group(n=40, H-Y stage 2.5-4.0), the non-depressed PD group(n=71) vs. the depressed PD group(n=16), the non-anxions PD group(n=62) vs.the anxions PD group(n=25). An ordinal regression model was used to investigate the correlations of WML with gender, age, Mini-Mental State Examination(MMSE)score, Unified Parkinson's disease Rating Scale-Ⅲ score(UPDRS-Ⅲ), Hamilton Rating Scale for Depression score(HAMD)and Hamilton Rating Scale for Anxiety score(HAMA). Results:Compared with the early PD group, the middle-advanced PD group showed that the WML were increased in lobe of brain(5.30±4.85 vs. 3.43±3.13, P<0.05), especially in the occipital lobe(0.48±0.99 vs. 0.11±0.31, P<0.05). There was no significant difference in the WML between the non-depressed/anxions and the depressed/anxions PD group.After being evaluated by the Scheltens scale, WML in periventricular hyperintensities(PVH)regions( OR=1.13, P<0.01), in brain lobe( OR=1.10, P<0.01)and in basal ganglia regions( OR=1.15, P<0.01)were correlated with age.WML in the brain besides the PV region were correlated with MMSE score( OR=0.68, P<0.01), especially in posterior horns( OR=0.60, P<0.01)and lateral ventricles( OR=0.68, P<0.05). WML in temporal lobe was correlated with MMSE score( OR=0.68, P<0.05). WML in brain lobe was correlated with H-Y stages( OR=2.10, P<0.05), especially in the occipital lobe( OR=3.33, P<0.05). WML in parietal lobe was associated with HAMD score( OR=1.13, P<0.05). WML in basal ganglia regions was related to diabetes( OR=6.34, P<0.05), especially in the putamen( OR=6.86, P<0.01). After being evaluated by the Fazekas scale, WML in PVH region( OR=1.16, P<0.01)and deep white matter hyperintensities( OR=1.13, P<0.01)were correlated with age.WML in PVH region were associated with MMSE score( OR=0.65, P<0.01). WML scores in PD patients had no correlation with gender, hypertension, coronary heart disease, hyperlipemia, UPDRS-Ⅲ score and HAMA score. Conclusions:The WML is present in PD patients, and it is correlated with age, diabetes, severity of disease, depression and cognitive function.

Objective To investigate the prevalence of depression in Parkinson's disease ( PD) patients, analyze the clinical features of depression in PD patients , and evaluate its impact on quality of life. Methods One hundred and ninety-five PD patients and 63 normol controls were recruited in this study.The detailed clinical information was documented.Unified Parkinson's Disease Rating Scale and Hoehn-Yahr stage were used to evaluate the severity of motor function impairment in PD patients.Hamilton Depression Rating Scale (HAMD) and Hamilton Anxiety Rating Scale were employed to assess the severity of depression and anxiety in PD patients.The 39-item Parkinson's Disease Questionnaire was applied to assess the quality of life.The cross-sectional data were calculated with SPSS 21.0 statistic software, and P ＜0.05 was considered statistically significant.Results The average score of HAMD was 8.74 ±5.51 in 195 PD patients.Depressive symptoms were found in 54.4%of the PD patients ( mild depression 48.7% and moderate depression 5.6%).Depression significantly impaired the quality of life in PD.Compared with PD without depression, PD with depression earned more scores in anxiety factor (4 (2, 5) vs 1(0, 2), Z＝ -8.69, P＝0.00), blocker factor (2 (1, 3) vs 0(0, 1), Z＝-7.95, P＝0.00), cognitive factor (1 (0, 2) vs 0(0, 0), Z＝-7.01, P＝0.00), sleep factor (2(1, 3) vs 0(0, 1), Z＝-6.42, P＝0.00) and despair factor (2 (1, 3) vs 1 (0, 1), Z＝-7.16, P＝0.00).There was no significant difference in day and night change (0(0, 0) vs 0(0, 0), Z＝-0.19, P＝0.85) and body weight (0(0, 0) vs 0(0, 0), Z＝-1.28, P＝0.20) between these two groups.The PD with depression obtained higher scores in total quality of life (30(22, 44) vs 14 (5, 24), Z＝-7.03, P＝0.00), motor function (6 (2, 13) vs 1 (0, 5), Z＝-3.67, P＝0.00), daily life ability (4 (1, 8) vs 1 (0, 4), Z＝-2.81, P＝0.01) , emotional health (5 (2, 11) vs 0 (0, 2), Z＝-5.82, P＝0.00), humiliation (2 (0, 5) vs 0 (0, 1), Z＝-3.10, P＝0.00), social support (0 (0, 1) vs 0 (0, 0), Z＝-2.86, P＝0.00), recognition function (4 (2, 6) vs 2 (0, 4), Z＝-2.87, P＝0.00), sociability(1(0, 3) vs 0(0, 1), Z＝-3.25, P＝0.00), and body pain (3 (1, 6) vs 1 (0, 2), Z＝-3.91, P＝0.00) than patients without depression.Conclusions Incidence of depression ( mainly mild ) in PD patients is high. Depressive symptoms significantly affect the quality of life of PD patients.

Objective:To investigate the grey matter alterations of Parkinson′s disease (PD) patients with and without sleep disorders, and to explore the relationship between different sleep-related problems and clinical variables as well as grey matter volume (GMV) in PD.Methods:Forty-six PD patients and 38 healthy controls (HCs) were recruited from January 2018 to December 2021 in the Department of Neurology, Beijing Hospital. PD patients were divided into PD with sleep disorders (PD-S, n=26) and PD without sleep disorders (PD-nS, n=20) subgroups (cutoff points of 82 for Parkinson′s Disease Sleep Scale or less than 5 for each item was considered as an indicator of substantial sleep disorder). The Mini-Mental State Examination (MMSE), the third part of the Unified Parkinson′s Disease Rating Scale (UPDRS-Ⅲ), Hamilton Rating Scale for Anxiety (HAMA), Hamilton Rating Scale for Depression (HAMD), Non-Motor Symptoms Questionnaire (NMSQ), and Parkinson′s Disease Questionnaire-39 (PDQ-39) were used to evaluate cognitive function, motor symptoms, anxious and depressive symptoms, non-motor symptoms, and the quality of life of the patients. Optimized voxel-based morphometry was applied to the magnetic resonance imaging brain images in all participants,and multiple linear regression analysis was used to test the correlation between GMV and sleep quality in patients with PD. Results:Compared with the HCs, PD-nS patients showed decreased GMV in bilateral limbic lobe, parahippocampal gyrus, amygdala, cingulate gyrus, hippocampus, right cerebellum, bilateral frontotemporal lobe, bilateral occipital lobe and the left parietal lobe. PD-S group exhibited reduced GMV in bilateral limbic lobe, parahippocampal gyrus, amygdala, right cerebellum, bilateral frontotemporal lobe and bilateral parietal-occipital lobe, compared to the HCs. Compared with PD-nS, PD-S patients revealed higher depressive (HAMD score: 12.19±5.59 vs 6.95±3.19, t=-4.01, P<0.001), anxious (HAMA score: 12.04±5.32 vs 7.25±4.68, t=-3.18, P=0.003), and non-motor symptoms scores (NMSQ score: 12.92±5.18 vs 9.90±4.10, t=-2.14, P=0.038), poorer quality of life (PDQ-39 score: 35.31±22.01 vs 22.40±9.00, t=-2.71, P=0.010), and reduced GMV in the left insula, frontal, and parietal lobe ( P<0.001, uncorrected, cluster>100). There was a marked relationship between sleep quality and the reduced GMV of the right medial temporal gyrus (β=0.006, 95% CI 0.002-0.010, P=0.003), left middle frontal gyrus (β=0.006, 95% CI 0.002-0.010, P=0.002), the right cerebellum (β=0.014, 95% CI 0.005-0.023, P=0.003), and the right medial occipital gyrus (β=0.017, 95% CI 0.011-0.024, P<0.001). Significant grey matter changes were associated with nocturnal restlessness, mainly within the left limbic lobe, bilateral occipital lobe, the right cerebellum, and parietal lobe (β=0.008, 95% CI 0.006-0.010, P<0.001). Furthermore, nocturia in PD was related to certain grey matter atrophy, including bilateral limbic lobe, the right inferior parietal gyrus, and bilateral frontal lobe (β=0.010, 95% CI 0.008-0.013, P<0.001). The symptom of daytime dozing was correlated with GMV reduction in the right occipital lobe, the left temporal lobe (β=0.014, 95% CI 0.010-0.019, P<0.001). There were also several compensatory brain regions, including bilateral frontal lobe, the left limbic lobe and cingulate ( P<0.001, uncorrected, cluster>60). Conclusions:Sleep disturbance is common in PD, which is related to the anxious and depressive symptoms, non-motor symptoms, and the quality of life. PD patients with different sleep disorders show grey matter alterations in severeal brain regions, which are associated with sleep quality, nocturnal restlessness, psychosis, and daytime dozing.

Objective:To investigate the characteristics and clinical related factors of Parkinson′s disease (PD) patients with subjective cognitive decline (SCD).Methods:Ninety-nine PD patients with normal cognitive function enrolled in Beijing Hospital from January to December 2018 were collected for the study. Patients with PD were divided into groups with ( n=57) and without ( n=42) SCD using the first question in Part 1 of the Unified Parkinson′s Disease Rating Scale (UPDRS). All patients were assessed by Montreal Cognitive Assessment (MoCA), modified Hoehn-Yahr grading, UPDRS, Hamilton Rating Scale for Depression (HAMD), Hamilton Rating Scale for Anxiety (HAMA), Parkinson′s Disease Sleep Scale, Ability of Daily Living Scale and 39-item Parkinson′s Disease Questionnaire (PDQ-39). Levodopa equivalent dose conversion was performed for patients taking anti-PD drugs. Patients′ self-reported years of formal education were collected. Results:The proportion of PD with SCD in this group was 57.58% (57/99). There were statistically significant differences in MoCA [28.00 (27.00, 29.00) vs 28.00 (27.00, 29.00) ,Z=-2.28, P=0.023], HAMD [6.00 (5.00, 8.50) vs 5.00 (2.00, 8.00), Z=-2.23, P=0.026], HAMA [7.00 (6.00, 11.00) vs 6.00 (3.00, 8.25) , Z=-2.70, P=0.007], PDQ-39-emotional health [2.00 (0, 5.00) vs 1.00 (0, 3.00), Z=-2.03, P=0.042] and PDQ-39-cognitive scores [4.00 (2.00, 5.00) vs 2.00 (0, 4.00), Z=-3.42, P=0.001] between PD with and without SCD groups. SCD was correlated with MoCA ( r=-0.23, P=0.022), HAMD ( r=0.23, P=0.025) and HAMA ( r=0.27, P=0.006) scores to varying degrees. When controlling for HAMD and HAMA scores, the correlation between SCD and MoCA scores ( r′=-0.18, P=0.084) was no longer existed. Conclusions:SCD is common in PD patients with normal cognitive function and is associated with poorer cognitive performance and more severe symptoms of depression and anxiety. In this group of patients, the relationship between SCD and affective symptoms may be greater than that of objective overall cognitive function, which is worthy of further studies.

Objective To investigate the characteristics and clinicopathology of v-raf murine sarcoma viral oncogene homolog Bl(BRAF)V600E mutations in papillary thyroid carcinoma(PTC)in Air Force flight personnel.Methods Data of cases and test results of BRAF V600E mutation were collected from Air Force aviators pathologically diagnosed with PTC.A univariate analysis of the relationship between BRAF V600E mutations and clinicopathologic features was performed.Results The overall rate of BRAF V600E mutations among 55 PTC flight crew members was 70.91％.The univariate analysis showed that the number of lymph node metastases in the BRAF V600E mutated group was larger than in the BRAF V600E unmutated group,and the proportion of BRAF V600E mutations in flight crews at intermediate risk of recurrence was higher than that in those at low risk of recurrence(P＜0.05).The presence or absence of BRAF V600E mutations did not affect the results of medical evaluation of PTC in flight personnel.Conclusion The rate of PTC BRAF V600E mutations in Air Force flight crews is similar to that of the general Chinese population.BRAF V600E mutations are associated with an increased number of lymph node metastases and risk of recurrence,and follow-up is recommended for flight personnel with PTC,especially those with BRAF V600E mutations.

Objective:To investigate the incidence of various non-motor symptoms (NMS) in early stage of Parkinson′s disease (PD) patients and the differences between the body-first and brain-first subtypes.Methods:A total of 121 patients with PD (Hoehn-Yahr stage 1-2) were recruited from PD Clinic, Department of Neurology, Beijing Hospital from January 2012 to January 2015. The general information and clinical features of the patients were collected. The minimal diagnostic criteria of parasomnias described in the International Classification of Sleep Disorders-Revised were used to diagnose rapid eye movement sleep behavior disorder (RBD).According to the sequence of RBD and motor symptoms, the patients were divided into 2 groups: body-first subtype and brain-first subtype. NMS was evaluated by the Non-Motor Symptom Questionnaire (NMSQuest). The clinical features and the incidence of various NMS were compared between the 2 groups. The Unified Parkinson′s Disease Rating Scale (UPDRS) was used to evaluate the severity of the disease, and its third part (UPDRS-Ⅲ) was used to evaluate the motor function of the patients. Hamilton Rating Scale for Depression (HAMD) and Hamilton Rating Scale for Anxiety (HAMA) were used to evaluate the depression and anxiety status of the patients. The sleep status of patients was assessed by Parkinson′s Disease Sleep Scale (PDSS). The quality of life of the patients was assessed by 39-item Parkinson′s Disease Questionnaire (PDQ-39).Results:Of all the patients, 49.59% (60/121) had the body-first subtype and 50.41% (61/121) had the brain-first subtype of PD. There was no significant difference in UPDRS-Ⅲ score between the 2 groups. The average number of NMS in all PD patients was 10.97±4.88. Body-first subtype patients had higher NMS incidence than brain-first subtype in difficulty in swallowing [46.7% (28/60) vs 23.0% (14/61), χ 2=7.507, P=0.006], nausea and vomiting [16.7% (10/60) vs 3.3% (2/61), χ 2=6.069, P=0.014], constipation [85.0% (51/60) vs 55.7% (34/61), χ 2=12.393, P<0.001], fecal incontinence [8.3% (5/60) vs 0 (0/61), χ 2=5.302, P=0.021], difficulty in remembering recent events [58.3% (35/60) vs 32.8% (20/61), χ 2=7.962, P=0.005], loss of interest [43.3% (26/60) vs 24.6% (15/61), χ 2=4.743, P=0.029], inattention [45.0% (27/60) vs 19.7% (12/61), χ 2=8.884, P=0.003], depression [55.0% (33/60) vs 34.4% (21/61), χ 2=5.181, P=0.023], intense vivid dreams [73.3% (44/60) vs 39.3% (24/61), χ 2=14.196, P<0.001] and restless legs [53.3% (32/60) vs 27.9% (17/61), χ 2=8.140, P=0.004]. The differences were significant. Body-first subtype and NMSQuest ( r=-0.489, P<0.001), UPDRS ( r=-0.189, P=0.038), HAMD ( r=-0.231, P=0.011), HAMA ( r=-0.298, P=0.001) and PDQ-39 scores ( r=-0.276, P=0.002) were negatively correlated. Body-first subtype and PDSS score was positively correlated. NMSQuest (Δ R2=0.265, P<0.001) was the main determinant of PDQ-39 score. Conclusions:PD patients are accompanied by various NMS, which is a major factor affecting the quality of life. Compared with brain-first subtype, body-first subtype might have more NMS burden and higher incidence rate in most NMS in early PD patients.