Objective To investigate the effects of low frequency repetitive transcranial magnetic stimulation (rTMS) applied to the supplementary motor area (SMA) of children with Tourette's syndrome (TS). Methods Thirty TS subjects less than 16 years old were treated with 1 Hz rTMS to the SMA at 110％ of the resting motor threshold (RMT) in 20 daily sessions,receiving 1200 pulses/day.Clinical assessment and physiological measures of the left and right RMT were conducted at different time points during the treatment. ResultsAfter 4 weeks of treatment,statistically significant reductions were observed in assessments with the Yale global tic severity scale (YGTSS) and in terms of clinical global impression (CGI).Symptomatic improvement was correlated with dramatic increases in both right and left RMTs. ConclusionApplication of 1 Hz rTMS to the SMA can improve the clinical symptoms of TS children.

Objective To study the effectiveness of personalized rehabilitation treatments based on threedimensional gait analysis (3DGA) for improving the walking function of children with cerebral palsy (CP).MethodsA total of 21 spastic CP children with diplegia or hemiplegia,IQ scores ＞60,and an average age of 8.5 years received 3DGA.They then received personalized rehabilitation treatment designed according to the 3DGA results.After four weeks of treatment the children accepted 3DGA again.Their gait descriptors before and after treatment were compared. ResultsAfter the personalized rehabilitation the subjects'clinical foot dorsiflexion angle,clinical popliteal fossa angle,walking velocity,stride length,step length,peak ankle dorsiflexion in prime stance,peaking ankle plantar flexion in last stance,peak back ground reaction forces (GRFs) and peak vertical GRF in stance all had improved significantly.The cadence,total support time,swing phase,initial double support time,peak knee extension in stance and the peak forward GRF were not,however,significantly different compared with before the personalized rehabilitation treatment.Conclusion Under the guidance of 3DGA,the walking function of spastic CP children improved significantly after 4 weeks of personalized rehabilitation treatment.3DGA can play an important role in formulating personalized rehabilitation protocols and guiding rehabilitation treatment for CP children.

Crimean-Congo Haemorrhagic Fever Virus(CCHFV)is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes,high fatality. The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment. In this research,the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus. Under an electron microscope,Virus-Like Particles (VLPs)with various size and morphology were observed in cytoplasmic vesicles in the infected cells.Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation,which was confirmed by Western blot analysis and immuno-electron microscopy(IEM).

Objective To investigate the characteristics and the correlative factors of constipation in Parkinson's disease (PD) patients.Methods The demographic information,clinical features and history of medications of 193 patients with idiopathic PD consulting in the outpatient department of the First Affiliated Hospital,Anhui Medical University were collected.Patients were evaluated using following scales:Unified Parkinson's Disease Rating Scale Ⅲ (UPDRS Ⅲ),Hoehn-Yahr stage,Bristol Scoring Scale,Cleveland Constipation Scoring Scale (CCS),Scale for Outcomes in PD-autonomic for Autonomic Symptoms,Simple Food Frequency Questionnaire,Hamilton Depression Scale (HAMD),Mini Mental State Examination.The patients were divided into constipation group and non-constipation group based on Rome Ⅲ Criteria for Diagnosis of Functional Constipation,and the correlative factors of constipation were compared and analyzed between the two groups.The severity of constipation and influencing factors were also compared between patients with early onset constipation (occurred before present of motor symptoms) and patients with late onset constipation (occurred after present of motor symptoms).The impacts of anti-PD medication adjustments on constipation were assessed by observing the alteration of constipation severity in 41 PD patients.Results The incidence of constipation was 56.4％ (109/193) in our cohort of PD patients,and 21.1％ (23/109) of constipation was severe according to the assessing by CCS.The spectrum of constipation symptoms included defecation straining (89.9％,98/109),poor stool output (67.9％,74/ 109),reduced stool frequency (63.3％,69/109) and dryness of stool (60.5％,66/109).The age,disease duration,scores of Hoehn-Yahr stage,UPDRS Ⅲ and HAMD,levodopa equivalent dose (LED),frequency of urination disturbance in constipation group were significantly higher than those in nonconstipation group,while the daily quantities of vegetable and water intake in constipation group were significantly lower than those in non-constipation group.Age and HAMD scores were the independent risk factors of constipation (OR=1.049,95％ CI 1.014-1.086,P=0.006;OR=1.316,95％ CI 1.185-1.461,P =0.000).Among the 109 constipation patients,the course of PD,scores of Hoehn-Yahr stage,UPDRSⅢ and LED were positively correlated with the severity of constipation (r =0.269,0.338,0.315,0.341,0.371,all P ＜ 0.05),with HAMD score being the independent risk factor of constipation severity (OR =1.175,95％ CI 1.044-1.322,P ＜ 0.05).The severity of constipation and risk factors of constipation in patients with early onset constipation were not distinct from those with late onset constipation.Conclusions The incidence of constipation increases with the increment of age,disease duration,Hoehn-Yahr stage,UPDRS Ⅲ scores,LED,HAMD scores,urination disorder severity and the decrement of daily water and vegetable intakes.The severity of constipation is positively related to the severity of motor symptoms,the daily dose of anti-PD drugs and depression levels.PD patients with early onset constipation are not distinct from the patients with late onset constipation in terms of severity and risk factors.The influences of anti-PD drugs on constipation are variable depending on the specific drug used and individual constitution.Individualized treatment regimes are proposed with respect to the management of constipation according to the specific risk factors in PD patients.

Objective To investigate the vein puncture assistive technology in the field of insufficient light conditions of auxiliary medical service staff quickly establish vein channel,improve the rescue efficiency and ensure the operational safety of application.Methods The night field training field,simulation of dark light intravenous therapy environment,select a project in Beijing City Corps training forces 429 soldiers as the research object,randomly divided into control group 214 cases,the use of military flashlight for intravenous infusion,215 cases in the observation group using the vein puncture assisted infusion,two group,the establishment of a single one-time successful puncture vein passage time and the incidence of adverse reactions.Results The simulation experiment in the observation group and the control group disposable puncture success rate were 95.8％,75.7％,the average puncture time was (66.38 ± 11.34)s,(89.94 ± 12.38) s,the incidence of adverse reaction were 3.88％,10.49％ The differences between the two groups were statistically significant (P ＜ 0.05).Conclusion Venipuncture assistive technology can improve the light environment,the success rate of vein puncture and strong practicability,high safety,wound treatment is suitable for various environments,fill the blank of vein puncture without light conditions,is of great significance to the security of military force.

[Objective] To observe the specific immune responses induced by the recombinant major outer membrane protein (rMOMP) vaccine against Chlamydia trachomatis E serotype in rhesus monkeys.[Methods] Six rhesus monkeys were equally divided into three groups:adjuvant and protein group vaccinated with purified rMOMP and Freund's adjuvants,adjuvant group immunized with Freund's adjuvants only,and control group immunized with phosphate buffer.All the rhesus monkeys were intramuscularly immunized in the triceps brachii for 3 times at a 2-week interval.Two weeks after the last vaccination,serum,vaginal wash and venous blood samples were collected from the rhesus monkeys,and lymphocytes were isolated from the blood samples.Enzyme linked immunosorbent assay (ELISA) was performed to determine the specific IgG antibody and interferon level in sera and secretory IgA (sIgA) level in wash samples,and methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferation of lymphocytes after stimulation with Chlaraydia trachomatis serotype E elementary bodies.Delayed hypersensitivity was observed in rhesus monkeys challenged by inactivated Chlamydia trachomatis serotype E elementary bodies.In vitro antibody neutralization assay was conducted with the serum from rhesus monkeys.Indirect immunofluorescenee was used to detect Chlamydia trachomatis in exfoliative vaginal cells from rhesus monkeys from week 1 to 10 after challenge with Chlamydia trachomatis.Data were statistically analyzed by using one-way analysis of variance and least significant difference (LSD) test with the SPSS 14.0 software.[Results] The adjuvant and protein group differed statistically from the adjuvant group and control group in the serum level of specific IgG antibody (1.718 ± 0.213 vs.0.841 ± 0.315 and 0.791 ±0.437,both P＜ 0.05),interferon ((1086 ± 121.730) ng/L vs.(409 + 53.440) ng/L and (162 ± 48.046) ng/L,both P＜ 0.05),lymphocyte proliferation index (7.012 ± 1.026 vs.4.473 ± 1.850 and 1A26 ± 1.104,both P＜0.01 ) and the diameter of nodus in delayed hypersensitivity assay ( ( 1 1 ± 2.134) mm vs.(3 ± 0.914) mm and 0,both P ＜ 0.01 ).After attack,the exfoliative cells kept positive for Chlamydia trachomatis in the adjuvant and protein group from week 1 to 5,and in the other 2 groups from week 1 to 10,but were negative in the adjuvant and protein group from week 6 to 10.[Conclusion] The rMOMP vaccine can induce a specific,protective,humoral and cellular immune response against Chlamydia tracbomatis in rhesus monkeys.

Objective To provide scientific basis for evaluating the efficiency of nursing service and reasonable allocation of human resources in blood collection by analyzing the time variation of visiting time in blood collection center. Methods Through the retrospective analysis of 2016 in different seasons and different blood centers working day and different time visits, obtains the variation of visits time;using a queuing analysis to evaluate different visits under the condition of nursing service efficiency and puts forward the scheme of optimal allocation of human resources. Results The blood center visits within one year of the first quarter of the lowest, the second quarter increased significantly, reaching the peak at the third quarter, the fourth quarter is relatively reduced; the work on Monday, Thursday, two or three high, five less;morning visits in different time was significantly higher than that in the afternoon, the morning peak on the 8:00-9:00 and 14:00-15:00 concentration in the afternoon peak; the situation of human resource allocation at this stage of blood centers, visits the peak season of nurse service intensity and low efficiency, the patients waiting for a long time, visits the low peak season, the nurse service intensity was low, idle for a long time, resulting in a waste of resources, quantitative configuration after the nurse service reasonable intensity, high work efficiency, effective treatment time of patients increased. Conclusions According to the law of time variation, the nursing service efficiency can be scientifically analyzed and the nursing manpower should be quantified to provide a scientific basis for hospital management.

Objective To investigate the influence of maternal high-protein diet during pregnancy and lactation on the growth of male rat offspring.Methods Female Wistar rats were mated with male rats and the pregnant ones were randomly assigned into three groups (n=10):Group Ⅰ,Group Ⅱ and Group Ⅲ.Three isocaloric purified diets,which were Diet Ⅰ (protein,14.0％;carbohydrate,69.3％;fat,16.7％),Diet Ⅱ (protein,24.0％;carbohydrate,59.3％;fat,16.7％) and Diet Ⅲ (protein,34.0％;carbohydrate,49.3％;fat,16.7％),were offered ad libitum to the rats in the three groups,respectively.On the 3rd day after birth,only three male rat offspring of each litter were kept.All male rat offspring at the age of 21 days were weaned onto the same normal diets until 77-day-old.Food intake (FI),body weight (BW),body length (BL) and visceral fat mass (VFM) of them were recorded.Blood lipids levels on the 21st,49th and 77th days after birth were detected.Weight gain (WG),food efficiency (FE) and percentage of VFM (VFM ％) were calculated to assess the growth of male rat offspring.One-way ANOVA,LSD or Tamhane T2 test was used for statistical analysis.Results (1) On the 3rd day after birth,male rat offspring in Group Ⅱ and Group Ⅲ had higher fast BW than those in Group Ⅰ [(9.77±0.81),(9.58±0.54) and (9.17±0.68) g;F=3.222,P=0.026].On the 7th,14th and 21st days after birth,male rat offspring in Group Ⅱ [(17.59± 1.24),(39.51 ±2.68) and (67.77±4.22) g] had higher BW than those in Group Ⅰ [(15.96±1.17),(35.35±4.11) and (63.43±3.98) g] and Group Ⅲ [(16.52±1.05),(37.06±3.27) and (64.43±3.81) g] (F=23.684,17.070 and 84.195,all P＜0.001).Male rat offspring's BL of Group Ⅱ were longer than those of Group Ⅰ on the 7th and 14th days after birth[(7.53±0.29) vs (7.53± 0.29) cm,(10.38 ±0.24) vs (9.99± 0.30) em,both P＜0.05].BL of Group Ⅲ was longer than that of Group Ⅰ,but shorter than that of Group Ⅲ on the 14th day after birth[(10.22 ± 0.25) vs (9.99± 0.30) cm,(10.22 ± 0.25) vs (10.38±0.24) cm,both P＜0.05].Male rat offspring's average WG from the 3rd to the 7th and the 7th to the 14th day after birth were higher in Group Ⅱ [(7.70±0.41) and (22.08±1.20) g] and Group Ⅲ [(7.00±0.40) and (20.75± 1.72) g] than in Group Ⅰ [(6.73±0.55) and (19.68± 1.73) g] (F=86.925 and 38.876,both P＜0.001),and the differences between Group Ⅱ and Group Ⅲ were statistically significant (both P＜0.05).(2) No significant differences in WG,FI and FE was observed among the three groups (all P＞0.05).Male rat offspring's BL on the 49th day after birth was longer in Group Ⅱ than in Group Ⅰ and Group Ⅲ [(22.03±0.26),(21.57±0.43) and (21.77±0.33) cm,F=3.222,P=0.026).VFM ％ of Group Ⅱ (3.87±0.32 and 5.13±0.32) and Group Ⅲ (3.90±0.27 and 5.15±0.33) on the 49th and 77th days after birth were higher than those of Group Ⅰ (3.50±0.34 and 4.68±0.38) (F=3.631 and 3.611,both P＜0.05).(3) Triglyceride (TG) level was higher in Group Ⅰ than in Group Ⅱ and Group Ⅲ on the 21st day after birth [(1.12±0.13),(0.89±0.10) and (0.97±0.12) mmol/L,F=7.283,P=0.004].However,Group Ⅱ and Group Ⅲ had a higher level of TG than Group Ⅰ on the 77th day after birth[(2.64±0.37),(2.43±0.32) and (1.90±0.21) mmol/L,F=12.321,P＜0.001].Conclusion Maternal high-protein diet can increase the birth weight of male rat offspring to a certain extent,which is influenced by carbohydrate content.Moreover,male rat offspring of dams fed with high-protein diet during pregnancy and lactation will have increased visceral fat accumulation and serum TG level during adulthood.

OBJECTIVETo explore the clinical features and genetic mutation in a family affected with non-syndrome X-linked intellectual disability (NS-XLID) using whole exome sequencing (WES).

METHODSMultiplex ligation-dependent probe amplification (MLPA) was applied to screen potential mutations of Fragile X syndrome (FXS). Whole exome sequencing (WES) and Sanger sequencing were screen for pathological mutations.

RESULTSFXS was excluded by MLPA analysis. WES has discovered in the proband an ARX gene mutation c.88G>T, which was confirmed by Sanger sequencing. Combining his clinical phenotype with information from the OMIM database, it was inferred that the ARX mutation probably underlies the NS-XLID in the proband. The same mutation was found in his mother and two uncles but not in his father and sister.

CONCLUSIONWES is capable of revealing the mutation underlying NS-XLID and can facilitate genetic counseling for the affected families.

Objective:To investigate the effect of conventional treatment plus long-term transcranial direct current stimulation (tDCS) on consciousness recovery in patients with minimally conscious state (MCS) after traumatic brain injury (TBI).Methods:A retrospective cohort study was conducted to analyze the clinical data of 60 patients with MCS after TBI admitted to Zhejiang Armed Police Corps Hospital from January 2019 to December 2020, including 38 males and 22 females, aged 19-60 years [(45.7±11.4)years]. Course of disease was 3-6 months [(4.6±0.9)months]. Of all, 30 patients received internal medicine, hyperbaric oxygen, rehabilitation and other conventional treatment (conventional treatment group), and 30 patients received tDCS stimulation on the basis of conventional treatment (tDCS treatment group). The tDCS stimulation contained 4 cycles for 28 days with each cycle lasting for 7 days (stimulation for 5 days, rest for 2 days). Coma recovery scale-revised (CRS-R) total score, brainstem auditory evoked potential (BAEP) score and clinical effictive rate (significantly effective+effective) were compared between the two groups before treatment, during 4 cycles of treatment and at 6 months and 12 months after treatment. Complications induced by tDCS were also evaluated.Results:There was no significant difference in CRS-R total score and BAEP score between the two groups before treatment (all P>0.05). CRS-R total score and BAEP score in tDCS treatment group were significantly higher than those in conventional treatment group during 4 cycles of treatment and at 6 months and 12 months after treatment (all P<0.05). CRS-R total score and BAEP score in both groups gradually increased during 4 cycles of treatment and at 6 months and 12 months after treatment (all P<0.05). The clinical effective rate in tDCS treatment group was 73% (22/30) when compared to 57% (17/30) in conventional treatment group ( P<0.05). In tDCS treatment group, 10 patients had local reversible slight redness at the cathodal position, while no other serious adverse effects, such as local burns, ulceration, exudation or epilepsy. Conclusion:Compared with conventional treatment, conventional treatment plus long-term tDCS can be more effective in improving the state of consciousness without serious adverse effects for MCS patients after TBI.