Objective To investigate the role of high-temperature requirement factor A2(HTRA2)in the mitochondrial unfolded protein response(mtUPR)model induced by the neurotoxin 1-methyl-4-phenylpyridinium(MPP+)and the compensatory effects of other proteases.Methods The mtUPR cell model was established by treating SH-SY5Y cells with MPP+.Cell viability was assessed using the Cell Counting Kit-8(CCK-8)assay,and mitochondrial morphological changes and intracellular reactive oxygen species(ROS)levels were examined under transmission electron microscopy and fluorescence microscope,respectively.Western blot(WB)was used to detect the expression levels of C/EBP Homologous Protein(CHOP),Heat shock protein family E member 1(HSPE1),YME1-like 1 ATPase(YME1L1),Caseinolytic mitochondrial matrix peptidase proteolytic subunit(CLPP),and Lon peptidase 1,mitochondrial(LONP1).Lentivirus-mediated knockdown(KD)of the HTRA2 gene in SH-SY5Y cells was performed.The mRNA and protein expression levels of HTRA2 were detected by quantitative real-time polymerase chain reaction and WB,respectively.After induction of mtUPR in HTRA2-KD SH-SY5Y cells with 400 μmol/L MPP?,cell viability and protein expression levels of HTRA2,YME1L1,CLPP,and LONP were evaluated using the CCK-8 assay and WB,respectively.Results mtUPR cell model was successfully established following treatment of SH-SY5Y cells with 400 μmol/L MPP+for 24 hours.Compared to the control group,there was no significant change in cell viability.Under electron microscopy,there were no remarkable alterations in the mitochondrial cristae and size in the mtUPR group.The average fluorescence intensity of ROS in the mtUPR group was 1.25±0.08 fold that of the PBS group,and the difference was statistically significant(P=0.001).The expression levels of CHOP,HSPE1,HTRA2,CLPP,and LONP were significantly higher in the mtUPR group than in the control group,which were 2.68±0.94、2.83±0.89、1.67±0.20,1.65±0.28,and 1.66±0.13 times that of control group,respectively(all P<0.05).The expression level of YME1L1 in the mtUPR group was 1.28±0.27 times that of the control group,with no statistical significance(P>0.05).After knockdown of the HTRA2 gene in SH-SY5Y cells,the expression levels of CLPP,YME1L1,and LONP1 increased to 1.46±0.79,1.41±0.12,and 1.32±0.25 times that of the empty virus group,respectively(all P<0.05).Following treatment with 400 μmol/L MPP+for 24 hours in HTRA2-KD SH-SY5Y cells,cell viability in HTRA2-KD group decreased to 88.4%±6.1%of the empty virus group(P<0.05).In the HTRA2 group,there were no significant changes in the expression levels of YME1L1 and LONP1 compared to the empty virus group(all P>0.05),while the expression level of CLPP increased to(1.88±0.62)times that of the empty virus group(P=0.033).Conclusion HTRA2 is an important mitochondrial protease in the MPP+-induced mtUPR response,and other mitochondrial proteases partially compensate for the function of HTRA2.

Objective To investigate the role of high-temperature requirement factor A2(HTRA2)in the mitochondrial unfolded protein response(mtUPR)model induced by the neurotoxin 1-methyl-4-phenylpyridinium(MPP+)and the compensatory effects of other proteases.Methods The mtUPR cell model was established by treating SH-SY5Y cells with MPP+.Cell viability was assessed using the Cell Counting Kit-8(CCK-8)assay,and mitochondrial morphological changes and intracellular reactive oxygen species(ROS)levels were examined under transmission electron microscopy and fluorescence microscope,respectively.Western blot(WB)was used to detect the expression levels of C/EBP Homologous Protein(CHOP),Heat shock protein family E member 1(HSPE1),YME1-like 1 ATPase(YME1L1),Caseinolytic mitochondrial matrix peptidase proteolytic subunit(CLPP),and Lon peptidase 1,mitochondrial(LONP1).Lentivirus-mediated knockdown(KD)of the HTRA2 gene in SH-SY5Y cells was performed.The mRNA and protein expression levels of HTRA2 were detected by quantitative real-time polymerase chain reaction and WB,respectively.After induction of mtUPR in HTRA2-KD SH-SY5Y cells with 400 μmol/L MPP?,cell viability and protein expression levels of HTRA2,YME1L1,CLPP,and LONP were evaluated using the CCK-8 assay and WB,respectively.Results mtUPR cell model was successfully established following treatment of SH-SY5Y cells with 400 μmol/L MPP+for 24 hours.Compared to the control group,there was no significant change in cell viability.Under electron microscopy,there were no remarkable alterations in the mitochondrial cristae and size in the mtUPR group.The average fluorescence intensity of ROS in the mtUPR group was 1.25±0.08 fold that of the PBS group,and the difference was statistically significant(P=0.001).The expression levels of CHOP,HSPE1,HTRA2,CLPP,and LONP were significantly higher in the mtUPR group than in the control group,which were 2.68±0.94、2.83±0.89、1.67±0.20,1.65±0.28,and 1.66±0.13 times that of control group,respectively(all P<0.05).The expression level of YME1L1 in the mtUPR group was 1.28±0.27 times that of the control group,with no statistical significance(P>0.05).After knockdown of the HTRA2 gene in SH-SY5Y cells,the expression levels of CLPP,YME1L1,and LONP1 increased to 1.46±0.79,1.41±0.12,and 1.32±0.25 times that of the empty virus group,respectively(all P<0.05).Following treatment with 400 μmol/L MPP+for 24 hours in HTRA2-KD SH-SY5Y cells,cell viability in HTRA2-KD group decreased to 88.4%±6.1%of the empty virus group(P<0.05).In the HTRA2 group,there were no significant changes in the expression levels of YME1L1 and LONP1 compared to the empty virus group(all P>0.05),while the expression level of CLPP increased to(1.88±0.62)times that of the empty virus group(P=0.033).Conclusion HTRA2 is an important mitochondrial protease in the MPP+-induced mtUPR response,and other mitochondrial proteases partially compensate for the function of HTRA2.

Many pathogenic genes have been identified in early-onset Parkinson′s disease, but the early-onset Parkinson′s disease with 22q11.2 deletion has not been reported in Chinese. A case of early-onset Parkinson′s disease with 22q11.2 deletion was confirmed by whole-exome sequencing-based copy number variation detection in Fujian Medical University Union Hospital. This article reports its clinical characteristics and discusses its pathogenesis, diagnosis and treatment management.

Objective:To investigate the clinical manifestations of familial amyloid multiple neuropathy (FAP) caused by Ala117Ser mutation, and to improve the clinical recognition of FAP.Methods:The clinical manifestations, electrophysiological examination, pathology and gene mutation characteristics of a case of FAP, who admitted to Fujian Medical University Union Hospital on November 25, 2014, were analyzed, and the literatures on the FAP cases caused by Ala117Ser mutation were reviewed and summarized.Results:The patient was a 59-year-old male from Fujian province. The first symptom was numbness in the extremities, followed by obvious autonomic nerve symptoms and motor disorder, and fatal cardiac dysfunction occurred in the later stage of the disease. The skin biopsy showed amyloidosis, and transthyretin gene analysis indicated the mutation of c.349G>T p.Ala117Ser. The clinical manifestations of FAP caused by Ala117Ser mutation reported in literatures are consistent with this case. And the reported FAP cases in China are concentrated in southern regions such as Fujian Province and Guangdong Province.Conclusions:Ala117Ser mutation in FAP patients is usually late onset and clinically manifested as multiple sensorimotor peripheral neuropathy, accompanied by prominent autonomic symptoms. The distribution of the patients has significant regional characteristics. Histopathological and genetic tests for the clinical diagnosis are of great significance.

Objective:To evaluate the sleep disorders in patients with Parkinson′s disease (PD) with different onset sides, and to analyze the correlation between PD kinesia-onset side and sleep disorders.Methods:A total of 658 patients with primary PD admitted to the Special Outpatient Department of Parkinson′s disease in Fujian Medical University Union Hospital from January 2015 to March 2021 were collected. According to the onset side of motor symptoms, they were divided into the left group (313 cases) and the right group (345 cases). The medical history collection and physical examination were conducted to evaluate the motor symptoms, non-motor symptoms [Non-Motor Symptom Scale (NMSS)], depression state and cognitive function of the patients. Parkinson′s Disease Sleep Sclale-2 (PDSS-2) and the Rapid Eye Movement Sleep Behavior Disorder Screening Questionnaire (RBDSQ) were used to evaluate and analyze their sleep status, and comparisons were made between groups. Binary multivariate Logistic regression analysis was used to access the risk factors associated with sleep disorders in Parkinson′s disease.Results:The scores of daytime fatigue [2.00(0, 4.00)] and unexplained limb pain [4.00(0, 4.00)] in NMSS assessment of PD patients in the left onset group were significantly higher than those in the right onset group [1.00(0, 3.00), Z=-2.545, P=0.001; 2.00(0, 4.00), Z=-2.797, P=0.005]. There was no significant difference in the total score of PDSS-2 between the two groups, but there were significant differences in limb restlessness, periodic limb activity, muscle spasm and early drowsiness between the two groups. In the evaluation of rapid eye movement sleep behavior disorder (RBD), the total score of RBDSQ in the left onset group [2.00(0, 4.00)] was significantly higher than that in the right onset group [1.00(0, 3.00), Z=-4.363, P<0.001]. The incidence of dream content, nocturnal behavior, nocturnal exercise, self-injury and bed partner in dream, abnormal behavior at night, nighttime awakening, dream memory and sleep disorder in the left onset group was also higher than that in the right onset group. In addition, binary multivariate Logistic regression showed that PD-related sleep disorders were associated with onset of advanced age ( OR=1.037, 95% CI 1.018-1.057, P<0.001), course of disease ( OR=1.014, 95% CI 1.010-1.018, P<0.001) and onset of abnormal postural gait ( OR=1.505,95% CI 1.058-2.141, P=0.023). RBD in patients with PD was associated with left onset ( OR=2.215,95% CI 1.395-3.515, P=0.001), advanced age onset ( OR=1.045,95% CI 1.019-1.072, P=0.001) and course of disease ( OR=1.014,95% CI 1.009-1.019, P<0.001). Conclusions:PD patients with left onset are more likely to have sleep disorders such as limb restlessness, periodic limb activity, muscle spasm and early drowsiness. At the same time, the incidence and severity of RBD in patients with left onset of PD are significantly higher than those of patients with right onset of PD. The onset side of motor symptoms of PD is an important factor affecting sleep disorders, and the onset of left side may be a risk factor for PD patients with RBD.

Objective:To explore the value of magnetic resonance diffusion kurtosis imaging (DKI) in the early diagnosis of Parkinson′s disease (PD).Methods:Thirty patients with early PD who were admitted to the Fujian Medical University Union Hospital From December 2016 to September 2017 were selected as case group, and 20 healthy persons in the same period were selected as healthy control group. 3.0 T GE magnetic resonance DKI was used to analyze olfactory related brain regions of the case group and the control group, the statistical differences were compared between the two groups in fractional anisotropy (FA), mean diffusion (MD), mean kurtosis (MK) values of olfactory cortex, and the correlation between MK values of olfactory cortex of the case group and age, disease course, Hoehn-Yahr (H-Y) stage, olfactory test, cognitive function evaluation was analyzed. Receiver operating characteristic (ROC) curve was used to determine the best diagnostic cutoff value of olfactory cortical MK in PD.Results:The left amygdala MK between the PD group (0.595±0.037) and the control group (0.647±0.091) showed statistically significant difference ( t=-2.183, P=0.037). ROC curve was drawn according to the MK value of the left amygdala of the PD group: the best diagnostic cutoff value was 0.597, the sensitivity was 65.0%, and the specificity was 52.2%. There was a statistically significant difference between the PD group and the control group in Montreal Cognitive Assessment (MoCA; 21.03±3.71 vs 24.25±1.65, t=-3.636, P=0.001) and Frontal Assessment Battery (FAB) scores (13.93±1.36 vs 15.00±1.25, t=-2.086, P=0.042), and there was negative correlation between MoCA, FAB scores and H-Y stage ( r=-0.548, P=0.007; r=-0.465, P=0.025). There was a positive correlation between MK value of the right direct gyrus of the PD group and MoCA evaluation results ( r=0.447, P=0.032). Conclusions:The left amygdala DKI can be used as a biomarker of PD in the early stage, which is helpful for the early diagnosis of PD. MoCA and FAB scales can be used as tools to monitor the progress of PD cognitive impairment. PD cognitive dysfunction may be related to impairment of frontal lobe function.

Objective To evaluate the efficacy of gait training and assessment system of Lokomat automatic robot (Lokomat robot) in patients with Parkinson's disease (PD).Methods Based on Hoehn-Yahr scale, 30 PD patients ranging from stage 2.5 to 3 were included and randomly assigned to Lokomat robot group ( n＝15) and control group ( n＝15).Lokomat robot system was employed in the training session of the Lokomat robot group, whereas patients in the control group were trained under auditory and visual guidance.Each training session lasted for 20 minutes, and repeated three days per week.Three motor assessments were performed before and after the four weeks training , including timed up and go test (TUGT), Unified Parkinson's Disease Rating Scale Ⅲ(UPDRS-Ⅲ) and three-dimensional gait analysis. Repeated measure analysis was performed under general linear mode , using SPSS 20.0.Results Gender, age, height and age of onset were matched in the Lokomat robot and the control groups .Scores of UPDRS-Ⅲ(Lokomat robot group , 23.46 ±2.72 vs 15.87 ±2.07; control group, 23.73 ±1.98 vs 18.07 ±0.80) and results of TUGT (Lokomat robot group, (15.42 ±5.59) vs (10.06 ±4.88) min; control group, (15.75 ± 4.67) vs (12.98 ±3.24) min) showed statistically significant differences before and after the gait training (UPDRS-Ⅲ, F＝258.598, P＝0.000; TUGT, F＝64.998, P＝0.000), and between the two groups (UPDRS-Ⅲ, F＝5.492, P＝0.026; TUGT, F＝6.522, P＝0.016).The step length (Lokomat robot group, (40.00 ±7.05) vs (52.70 ±7.62) cm; control group, (39.16 ±4.52) vs (46.72 ±7.29) cm), stride length (Lokomat robot group, (76.03 ±12.50) vs (90.60 ±12.46) cm; control group, (77.25 ± 8.07 ) vs (88.21 ±8.17) cm), walking pace ( Lokomat robot group, (67.16 ±12.79) vs (83.72 ± 10.96) m/min; control group, (65.35 ±11.56) vs (77.18 ±10.60) m/min), and total supporting phase (Lokomat robot group, 62.31% ±3.32% vs 56.05% ±3.98%; control group, 62.52% ±3.73% vs 57.96%±3.51%) showed significant improvement after training ( step length, F＝90.866, P＝0.000;stride length, F＝218.152, P＝0.000; walking pace, F＝172.236, P＝0.000; total supporting phase , F＝197.945, P＝0.000).Meanwhile, these improvements were more significant in the Lokomat robot group than the control group ( step length, F＝5.853, P＝0.022; stride length, F＝4.346, P＝0.046;walking pace, F＝4.904, P＝0.035; total supporting phase, F＝4.845, P＝0.036).No significant difference in step frequency was found before and after gait training.Conclusion Both gait trainings improved walking ability in PD patients , and Lokomat robot system guided training showed more obvious improvement than the traditional training under hearing and visual cue.

Objective To investigate the diagnostic value of quantitative detection of α-synuclein and DJ-1 protein in saliva for Parkinson Disease. Methods Twenty seven patients diagnosed with primary Parkinson's disease and 27 healthy controls were studied.The clinical data of all subjects were collected.Each participant received a disease evaluation including Hohn-Yahr stage, unified Parkinson Disease Rating Scale (UPDRS)-Ⅱ / Ⅲ, 12 Item odor identification test from Sniffin'Sticks (SS-12), Montreal cognitive assessment (MoCA), Mini Mental State Examination (MMSE).α-syn and DJ-1 protein in saliva were examined by using enzyme linked immunosorbent assay (ELISA). The statistical analysis was used to test the difference in these two protein levels between patient and control groups and the correlation with age, gender and course of disease. Results There were significant changes in mean concentration of salivary α-syn (1269.02±16.09 pg/mL、1350.51±25.79 pg/mL,P=0.010) and DJ-1 protein (6.07±3.23 ng/mL、8.43±4.33 ng/mL,P=0.027) between patient and control groups. The sensitivity of α-syn and DJ-1 protein levels in PD diagnosis was 55.56% and 77.8%,and the specificity was 89.19% and 55.6%.The area under the ROC curve in finding the PD of α-syn and DJ-1 was 0.671 and 0.649, respectively. In Parkinson disease group, age, gender, UPDRS-Ⅱ / Ⅲ, Hohn-Yahr stage, SS-12, MMSE and MoCA of Parkinson's disease group were not related to the concentration of α-syn and DJ-1 protein in saliva (P>0.05). Conclusion The detection of α-syn and DJ-1 protein levels in saliva may be an auxiliary tool for diagnosis of PD.

Objective To explore the incidence of olfactory dysfunction in patients with Parkinson disease and the characteristic as well as its possible influencing factors. Methods The SS-12 was used to evaluate the olfactory function of 106 patients with Parkinson's disease and 110 healthy volunteers. The data was then compared between the two groups to investigate the correlation of olfactory function with age, gender, education, smoking, disease duration, Hohn-Yahr stage, UPDRSⅢscores, the dosage of levodopao and olfactory scores. Results Mean identification scores were significantly lower in patients(5.97 ± 2.27)than in controls(8.04 ± 2.00)(t=7.108, P=7.108). Parkinson's disease group did worse than the control group in identifying some odors including peppermint, bananas,liquorice,coffee,pineap?ple,rose and fish (P<0.05). Receiver Operating Characteristic curves revealed that 7.5 was the best olfactory dysfunc?tion diagnostic boundary value with 67.3%sensitivity and 73.6%specificity. Thus, the incidence of olfactory dysfunc?tion in Parkinson's disease was 73.6%. Correlation analysis showed that gender(rs=0.243, P=0.012), education(rs=0.208, P=0.032) and smoking(rs=0.279, P=0.279) were associated with odor identification ability, while age, disease du?ration, Hohn-Yahr stage, UPDRSⅢscores and the dosage of levodopa were not related with odor identification abilityin Parkinson disease (P>0.05). Conclusion Olfactory dysfunction occurs in Parkinson disease with an hign incidence rate. Olfactory function has nothing to do with disease duration, Hohn-Yahr stage, UPDRSⅢscores and the dosage of levodopa in Parkinson disease.

Objective To systematically assess the efficacy of memantine on moderate to severe Alzheimer's disease (AD).Methods With the evaluation method of the Cochrane system,searches were made in the Cochrane Library,MEDLINE,Embase,Forest Laboratories,CNKI,Wanfang Data,and VIP Data up to February 2013 for double blind,randomized,and placebo-controlled trials (RCTs) evaluating the efficacy of memantine for moderate to severe AD.A meta-analysis of included clinical trials was conducted using the Revman 5.2 software to evaluate the efficacy of memantine on overall clinical status,cognitive function activities of daily living,and behavioral and psychological disturbances.Results A total of 8 RCTs were included (2 527 patients with moderate to severe AD).Results of the meta-analysis showed that,for patients with moderate to severe AD,memantine had better efficacy than placebo on overall clinical status,cognitive function,and activities of daily living (MD=-0.24,95％CI:0.340.15;SMD=-0.26,95％CI:-0.340.18;SMD=-0.13,95％CI:-0.21-0.05),but there was no significant difference in efficacy on behavioral and psychological function between memantine and placebo (P =0.08).Analysis of subgroups showed that memantine had better efficacy than placebo on cognitive function in moderate AD patients (SMD =-0.22,95％CI:-0.37 0.06) and on overall clinical status,cognitive function,and activities of daily living in severe AD patients (MD-0.29,95％CI:-0.40 0.18;SMD=-0.31,95％CI:0.46-0.15;SMD=-0.16,95％ CI:-0.25 0.06;MD=-3.13,95％ CI:-4.88-1.39;respectively).Conclusions Memantine has efficacy on overall clinical status,cognitive function and activities of daily living in patients with moderate to severe AD,especially in patients with severe AD.