Correlations of transcranial sonography of substantia nigra characteristics with MRI iron deposition on substantia nigra in patients with Parkinson disease

Objective To explore the correlations of transcranial sonography of substantia nigra(SN-TCS)characteristics with MRI iron deposition on substantia nigra in patients with Parkinson disease(PD).Methods Data of SN-TCS and craniocerebral MRI in 120 PD patients were retrospectively analyzed.The patients were divided into iron deposition positive group(positive group,n=46)and iron deposition negative group(negative group,n=74)according to quantitative susceptibility mapping(QSM)value.Then parameters of SN-TCS and MRI were compared between groups(both P<0.05),and correlation analysis were also performed.Results The proportion of high echo positive,strong echo area and QSM value of substantia nigra,as well as of hyper-substantia nigra area/midbrain area(S/M)in positive group were all higher than those in negative group(all P<0.001).No significant difference of midbrain area was found between groups(P>0.05).Strong echo area of substantia nigra and S/M based on SN-TCS were both low-medium positively correlated with substantia nigra QSM value showed on MRI(r=0.497,0.529,both P<0.001).Conclusion SN-TCS characteristics of PD patients were correlated with MRI iron deposition on substantia nigra,among which strong echo area and S/M were valuable for evaluating iron deposition on substantia nigra.

Prognosis Factors of Urinary Quality of Life in Parkinson Disease

Introduction: Quality of life in Parkinson disease is not necessary linked to motor symptoms. It’s correlated of overactive bladders and prostatic symptoms. Prognosis factors of urinary quality of life are unknown. Objectives: Our study aims to find prognosis factors of quality of life associated specifically of urinary disorders in PD. Patients and methods: We conducted a transversal, analytic and descriptive study in Physical Medicine, and Neurology departments, Fann Teaching Hospital, Dakar and included patients followed for treatment of PD. Urinary disorders, quality of life and functional autonomy were assessed respectively by Urinary Symptoms Profile (USP), Qualiven Short Form, Schwab and England Scale. Results: 38 patients presented PD, with a mean age of 60.89 ± 13.6 years and sex-ratio of 2.45. Mean duration of PD was 3.1 ± 2.9 years. Urinary disorders were found in 47.36% and dominated by incontinence (88.88%) and overactive bladders (88.88%) which were minor in more than 55%. Quality of life was impaired in 88.88% of cases. Minor forms (43.75%) were predominant and constraint dimension (50%) was the most severely altered. Prognosis factors for quality of life of urinary disorders were PD stages (0.046) and functional autonomy (0.042). Discussion and conclusion: Urinary disorders in PD are common, impaired quality of life, especially the constraint dimension, depends on stage of evolution of PD and functional autonomy of patients.

IMAGING CHARACTERISTICS OF RAT MODELS OF PARKINSON DISEASE

Imaging technologies are utilized to study the brain morphology and the functions of rat models of Parkinson disease （PD）. Magnetic resonance imaging （MRI） and magnetic resonance angiography （MRA） are used to obtain morphological imaging data. Functional imaging data, such as the spectrum and blood flow changes are obtained by proton magnetic resonance spectroscopy （1H-MRS） and CT perfusion （CTP）. Results show that PD rat models have no characteristic morphological imaging abnormalities, but exist regional cerebral blood flow （CBF） reductions and spectral changes in the striatum.

Experimental Study of Serum Substantia Nigra Neuron Autoantibody and Its Effect in Parkinson Disease Patients

To investigate the serum substantia nigra neuron autoantibody and its effect in the patients with Parkinson disease (PD), substantia nigra slices and a rat model of injection of serum from PD patients in unilateral side substantia nigra were applied. The results showed that the positive rate of substantia nigra neuron autoantibody in PD patients was significantly higher than in the healthy control group (36.67 % vs 6.67 %, P <0.01), but no significant difference was found between PD group and myasthenia gravis (MG) group (26.67 %, P >0.05). The sera from PD patients positive for substantia nigra neuron autoantibody could decrease the number of the dopaminergic neurons more seriously than those from MG and the healthy once respectively (both P <0.01). The results suggested that the immunological mechanism might partly play a role in the development of PD.

Experimental Study on the Protective Effect of Puerarin to Parkinson Disease

The protective effect of puerarin on the Parkinson disease (PD) mice with decreased estrogen level was investigated in order to develop a new potential medicine as a substitute for estrogen for preventing and treating PD. By using immunohistochemical method of avidinbiotin peroxidase complex (ABC), the distribution of the cells positive for tyrosine hydroxylase (TH) and fibres in the substantia nigra of the mouse were observed. These mice were divided into three groups randomly: group A , normal-female-mouse models; group B containing three subgroups, B1 (normal saline) , B2 (estrogen), B3 (puerarin); group C containing three sub groups, C1 (normal saline), C2 (estrogen), C3 (puerarin). By using TUNEL the numbers of apoptosis cells in every visual field was counted and the difference between the experimental group and control group was compared. The results showed the numbers of the cells positive for TH were more and the numbers of apoptosis cells were less in the normal-female-mouse models group than in the group of model made after ovar-iosteresis and the group of model made before ovariosteresis (P<0. 05), respectively. However, there was no significant difference, between the group given estrogen/puerarin and the controls, and between the group given estrogen and given puerarin. (P>0. 05). It was suggested that puerarin may have protective effect on the nigral neurons to PD. Moreover, the protective effect might serve as a surrogate of estrogen and be associated with the apoptosis.

Evaluation of cardiovascular risk in patients with Parkinson disease under levodopa treatment

BackgroundLevodopa 是在有 Parkinson 疾病(PD ) 的病人的中间的治疗的不可缺少的选择。因为 L-dopa 治疗被显示增加浆液 homocysteine 层次，为心血管的混乱的一个著名风险因素，有在 L-dopa 治疗下面的 PD 的病人将在为未来的增加的风险心血管的事件。这研究的目的是在 levodopa treatment.MethodsThe 学习下面与 PD 在病人评估心血管的风险人口在 L-dopa 治疗下面与自发的 PD 由 65 个病人组成了。控制组包括了 32 年龄，性匹配没有认知衰落的个人。Echocardiographic 大小，浆液 homocysteine 层次和主动脉的有弹性的参数与 PD 和 controls.ResultsAs 在病人之间被作比较 L-dopa 治疗的一个期望的特征， Parkinson 组有显著地更高的 homocystein 层次(15.1 ±；3.9 µ； mol/L 对 11.5 ±；3.2 µ； mol/L， P = 0.02 ) 。当时，大动脉的膨胀性在有 PD 的病人是显著地更低的与控制相比(4.8 ±；1.5 dyn/cm 2 对 6.2 ±；1.9 dyn/cm 2, P = 0.016 ) 。另外，有 PD 的病人有更高大动脉的紧张和大动脉的僵硬索引(13.4%±；6.4% 对 7.4%±；3.6% ， P <；0.001 和 7.3 ±；1.5 对 4.9 ±；1.9， P <；0.001 分别地) 。而且，浆液 homocysteine 层次被发现断然与大动脉的僵硬索引被相关，在浆液 homocysteine 的大动脉的膨胀性和层次之间有否定关联(r = 0.674， P <；0.001；r =−； 0.602， P <；0.001，分别地) 有在 L-dopa 治疗下面的 PD 的 .ConclusionsThe 病人增加了大动脉的僵硬并且与健康个人相比损害了心脏舒张的功能。提高的浆液 homocysteine 层次可以是可能的 pathophysiological 机制。

C.elegans as a model system for Parkinson disease

Parkinson disease(PD) is characterized by the selective loss of dopaminergic neurons in the substantia nigra. Although investigation in mammalian animal models of PD has enhanced our understanding of PD, the complexity of the mammalian nervous system and our inability to visualize DA neurons in vivo restricts the advances in elucidating the molecular mechanisms of PD. Conservation between C. elegans and mammals in genomic, biosynthetic and metabolic pathways as well as the advantages of observing DA neurons morphology in vivo and the ease of transgenic and genetic manipulation make C. elegans an excellent model organism for PD.

Expression of calbindin D28K in substantia nigra of model rats with Parkinson disease

BACKGROUND： Previous researches suggested that expression level of calbindin D28K mRNA decreased in substantia nigra （SN） of model rats with Parkinson disease （PD）, and this might be related to the decrease of anti-degeneration potentials of dopaminergic neurons. OBJECTIVE： To observe expression changes of calbindin D28K in SN dopaminergic neurons during their degeneration and death in midbrain of PD model rats. DESIGN： Completely randomized grouping design SETTING： Department of Neurobiology, Xuzhou Medical College MATERIALS＂ A total of 92 healthy male SD rats, with the age of 3 months, weighing 200-250 g, were selected from Experimental Animal Center of Xuzhou Medical College [certification： SCXK （su） 2003-0003]. Calbindin D28K（CB）, tyroxine hydroxylase （TH）, ABC kit, 6-hydroxydopamine （6-OHDA） and Nissl dyes were provided by Sigma Company, and sheep serum was provided by Beijing Zhongshan Company. METHODS： The experiment was carried out in the Neurobiological Center of Xuzhou Medical College from October 2003 to October 2004. ① With lot method, rats were divided into blank control group （n=28）, experimental control group （n=-28） and experimental group （n=36）. Rats in experimental group were injected with 6-OHDA at right corpus striatum for PD modeling; rats in experimental control group were injected with saline at the same site; rats in blank control group did not give any injections.② On the 7^th, 14^th, 21^st and 28^th days, SN segments on right midbrain from every 5 rats in experimental group were fixed, embedded with paraffin and cut into successively coronary pieces. Rats in other two groups were treated with the same methods and then stained with Nissl to show neuronal form. Meanwhile, CB and TH antibodies staining with immunohistochemistry were used to show CB containing dopaminergic neurons and dopaminergic neurons, and cells were calculated and observed under optic microscope. ③ On the 14^th and 28^th days, every 4 rats in experimental group and every 4 rats in control group were selected to obtain their brains and separate SN on the injured side. Western blot was used to detect expression of calbindin D28K, protein band was scanned with imaging equipment, and data were analyzed with LabWorks software. MAIN OUTCOME MEASURES：① On the 7^th, 14^th, 21^st and 28^th days, Nissl staining results of SN neurons and immunohistochemical staining results of CB and TH antibodies; ② On the 14^th and 28^th days, Western blot results of calbindin D28K in SN neurons, RESULTS; Among 92 rats, 2 rats in experimental group died after 1 day due to 6-OHDA injection and other 90 rats were involved in the final analysis. ①Nissl staining results： On the 7^th day of 6-OHDA injection, most neuronal somas on right SN pars compacta were shown as deep pycnosis or lysis breakage; on the 14^th and 21^st days, amount of neurons was decreased remarkably; on the 28^th day, most neurons in SN pars compacta disappeared. ② Results of immunohistochemical staining： Amount of positive neurons of calbindin D28K in right SN pars compacta was not changed on the 7^th day after 6-OHDA injection; on the 14^th day, the amount was higher in experimental group than that in both control groups （P 〈 0.01） and was decreased till the 21^st day, but it was still higher than that in the two control groups （P 〈 0.05）; on the 28^th day, positive neurons of calbindin D28K nearly disappeared, and the amount was lower than that in the two control groups （P 〈 0.01）. In addition, on the 7^th day after 6-OHDA injection into corpus striatum, positive TH neurons decreased 24% in right SN, and there was significant difference from that in control groups; on the 14^th, 21^st and 28^th days, positive TH neurons decreased 37%, 46% and 64%, respectively. ③ Western blot results： On the 14＇h day after 6-OHDA injection into corpus striatum, expression of calbindin D28K in right SN was higher in experimental group than that in both control groups （P 〈 0.05）; however, on the 28^th day, the expression was lower than that in the two control groups （P 〈 0.01 ）. CONCLUSION ： During degeneration and death of dopaminergic neurons, CB expression in SN pars compacta increases firstly and decreased gradually.

Tetramethylpyrazine analogue T-006 promotes clearance of alpha-synuclein by enhancing proteasome activity in Parkinson disease models

OBJECTIVE To investigate the effects of T-006(tetramethylpyrazine derivative)in promotingα-Synuclein(α-Syn)degradation and evaluated the neuroprotective effects in cellular and animalα-Syn model of Parkinson disease(PD).METHODS The inducible PC12 cells overexpressingα-syn and the homozygous transgenic(Tg)mice expressing A53T humanα-syn were used to evaluate the neuroprotective effects of T-006.For cellular study,MTT,Western blotting,proteasomal activity assay and qRT-PCR were applied to analyze the pharmacological effects and underlying mecha⁃nisms.The gene knock-down and overexpression approaches were used to dissect the molecular signaling pathways.For animal study,ten-month-old homozygousα-Syn Tg mice were treated with T-006(3 mg·kg-1)daily by gavage for four weeks.The Western blotting,immunohistochemistry and behavioral tests were applied to determine the neuropatho⁃logical changes.RESULTS T-006 promoted the degradation of WT and mutantα-Syn in PC12α-Syn inducible cells via an ubiquitin-proteasome system(UPS)dependent and autophagy-lysosome pathway independent manner.The mecha⁃nism of action involved the upregulation of 20S proteasome subunit LMP7 expression,which leads to activation of the chymotrypsin-like proteasomal activity for protein degradation.Mechanistically,we demonstrated that T-006 activated PKA/Akt/mTOR pathway upstream for LMP 7 up-regulation and UPS activation.Finally,we illustrated that T-006 promoted both Triton-soluble and-insoluble forms ofα-syn and protected againstα-Syn-induced neurotoxicity in A53Tα-Syn Tg mice.CONCLUSION T-006 is a potent UPS activator which promotes the degradation of pathogenic proteinα-Syn in cellular and animal PD models.Our study thus high-lights the therapeutic potential of small molecular UPS activator like T-006 in the treatment of PD and related conditions.

A small-molecule activator of ULK1 that induces cytoprotective autophagy for Parkinson disease treatment

OBJECTIVE To discover a small-molecule activator of ULK1 for Parkinson disease treatment and exploreits potential mechanisms.METHODS Candidate ULK1 activator was found by using structure-based design and high-through put screening,then modified by chemical synthesis and screened by kinase and autophgic activities.The amino acid residues that key to the activation site of the best candidate ULK1 activator(BL-918) were determined by site-directed mutagenesis,as well as in vitro kinase assay,ADP-Glo kinase assay and surface plasmon resonance(SPR) analysis.The mechanisms of BL-918 induced cytoprotective autophagy were investigated by electron microscopy,fluorescence microscopy,Western blotting,co-immunoprecipitation assay,si RNA and GFP-LC3 plasmid transfections.The therapeutic effect of BL-918 was determined by MPTP-mouse model,including behavioral tests,the levels of dopamine and its derivatives,as well as immunofluorescence and Western blotting.The toxicity of BL-918 was assessed by blood sample analysis and hematoxylin-eosin staining.RESULTS We discovered a small molecule(BL-918) as a potent activator of ULK1 by structure-based drug design.Subsequently,some key amino acid residues(Arg18,Lys50,Asn86 and Tyr89) were found to be crucial to the binding pocket between ULK1 and BL-918,by site-directed mutagenesis.Moreover,we found that BL-918 could induce autophagy via the ULK complex in neuroblastoma SH-SY5Y cells.Intriguingly,this activator displayed a cytoprotective effect on MPP+-treated SH-SY5Y cells,as well as protected against MPTP-induced motor dysfunction and loss of dopaminergic neurons by targeting ULK1-modulated autophagy in mouse models of PD.CONCLUSION We discovered a novel ULK1 activator(BL-918) that potently activated ULK1.This activator could induce cytoprotective autophagy via the ULK1 complex in SH-SY5Y cells,and also exerted its neuroprotective effects by targeting ULK1-modulated autophagy in a MPTP-induced PD mouse model,which may serve as a candidate drug for future PD therapy.

Magnetic Resonance Imaging of Transplanted Neural Stem Cells in Parkinson Disease Rats

Summary： In this study we implanted magnetically labeled neural stem cells （NSCs） in PD rats and then monitored their survival and migration in the host brain by magnetic resonance imaging （MRI）. The mesencephalic NSCs were obtained from the brain of SD rats. Superparamagnetic iron oxide （SPIO） was transferred to NSCs by Lipofectamine transfection. Eighteen PD lesioned rats were selected for transplantation by evaluation of their rotational behavior in response to amphetamine and randomly assigned to 3 groups, i.e., sham group, PBS group and NSCs transplanted group, with 6 rats in each group. MR scanning was performed at 1, 2, 4, 6, 8 and 10 week（s） following transplantation. At the meantime, rotational behavior was assessed in each group. Our results showed that SPIO particles were clearly visible with Prissian blue staining in neurospheres and cells derived from NSCs. The rotational behavior of the NSCs transplanted group was remarkably improved compared with that of sham group and PBS group （P〈0.05）. In vivo MR tracking of NSCs showed that SPIO labeling led to a strong susceptibility change of signal 1 week after transplantation on T2 weighted images. And a large circular hypointense signal appeared in the transplanted area on T2＊ gradient echo images Ten weeks following transplantation, the hypointense signal on T2 weighted and T2＊ gradient echo images was still displayed. It is concluded that SPIO particles could label NSCs effectively, and MRI detection of SPIO labeled cells is a promising method and novel approach to analyzing the NSCs following transplantation in the treatment of PD.

Reassessment subacute MPTP-treated mice model of Parkinson disease

OBJECTIVE(1) To estimate the value of the subacute MPTP mouse model in aspects of behavioral performance,biochemical changes and pathological abnormalities.(2) To find effective positive drugs.METHODS Male C57 BL/6 mice were injected with MPTP(30 mg·kg^(-1)·d^(-1),ip) for 5 consecutive days.Three days before MPTP injection,the mice were orally administered selegiline(3 mg·kg^(-1)·d^(-1)),pramipexole(3 mg·kg^(-1)·d^(-1)),or medopar(100 mg·kg^(-1)·d^(-1)) for 18 d.Behavioral performance was assessed in the open field test,pole test and rotarod test.Neurotransmitters in the striatum were detected using HPLC.Protein levels were measured by Western blot.Pathological characteristics were examined by immunohistochemistry.Ultrastructure changes were observed by electron microscopy.RESULTS The subacute MPTP treatment did not induce evident motor defects despite severe injuries in the dopaminergic system.Additionally,MPTP significantly increased the α-synuclein levels and the number of astrocytes in the striatum,and destroyed the blood-brain barrier(BBB) in the substantianigra pars compacta.Both selegiline and pramipexole were able to protect the mice against MPTP injuries.CONCLUSION The subacute MPTP mouse model does not show visible motor defects;it is not enough to evaluate the validity of a candidate just based on behavioral examination,much attention should also be paid to the alterations in neurotransmitters,astrocytes,α-synuclein and the BBB.In addition,selegiline or pramipexole is a better choice than medopar as an effective positive control for the subacute MPTP model.

Metabolic aspects behind ageing and neurodegeneration reveal new ways forward for therapy in motorneurone(ALS) and Parkinson diseases

Human beings evolved to run over a relatively short evolutionary time scale, ~1 million years, 2-3 million years ago, markedly increasing metabolic rate and VO2max, compared with other primates, while increasing brain size,and lifespan. Ageing leads to precise declines in performance and metabolism(VO2max): are there links with ageing-related diseases? Glucocerebrosidase(GCase;GBA1 subtype) mutations are the most common cause of Parkinson Disease,where there is lysosomal disruption and a reciprocal feedback between glucosylceramide and a-synuclein. We have shown that GBA2 is elevated and, using metabolomics, that ceramide and glucosylceramide levels are critically modified presymptomatically and at early stage in the spinal cord of superoxide dismutase1 mutant mice(SOD1G86R, ALS model),and lipid metabolism is massively changed at end stage disease. Modification of glucosylceramide synthase(GCS), and GCase activites shows that inhibiting GCS is deleterious and inhibiting GCase is beneficial to both neuromuscular junction function in sciatic nerve crush, and also grip strength and survival in the SOD1G86R model. Ambroxol(3 mmol·L-1 in drinking water), a glucocerebrosidase chaperone, accelerated recovery of NMJ function in sciatic nerve crush, and ameliorated grip strength and survival in the SOD1G86R model. Ambroxol is phase 2 ready in ALS and starting phase 3 in Parkinson disease.

Inhibition of miR-873 provides therapeutic benefit in lipopolysaccharide-induced Parkinson disease animal model

OBJECTIVE Neuroinflammation plays a critical role in neurodegenerative disorders,although the inflammation may not the initiating factor.Parkinson disease(PD)is characterized pathologically by the accumulation of alpha synuclein(α-syn)and the loss of the dopamine(DA)neurons in the substantia nigra(SN),which has been reported to be induced by the stereotaxic injection of lipopolysaccharide(LPS)to the SN region in rodents.This study is to investigate the therapeutic benefit of the inhibition of miR-873 in PD.METHODS Rats received the right-unilaterally injection with concentrated LV-sponge or LV-EGFP 3 d before LPS treatment,7 or 14 d after LPS treatment.The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 21 d after LPS injection.The regulation of miR-873 on the genes related with cholesterol transport and inflammation was assayed in SH-SY5Y cells and U251 cells.RESULTS TLR4-My D88 signaling pathway was involved the regulation of miR-873 by LPS.The luciferase assay showed that HMGCR,ABCA1 and A20 were down-stream genes of miR-873.The transfection of miR-873 decreased the cholesterol levels in cell membrane,but increased in lysosome in SH-SY5Y cells.Compared with the control SH-SY5Y cells,cholesterol levels were higher in lysosome withα-synuclein overexpression or LPS treatment.The transfection of miR-873 increased theα-syn levels in lysosome in cel s withα-synuclein overexpression.The loss of dopaminergic neuorns induced by LPS was significantly respectively decreased by 22.8%,35.6%and 57% after the inhibition of miR-873 at 3 d before LPS treatment,7 or14 d after LPS treatment.Compared with LPS-treated group,the number of the rotation of rats was decreased by 60.4%,33.5%and 13.2%after the inhibition of miR-873 at 3 d before LPS treatment,7or 14 d after LPS treatment.The inhibition of miR-873 significantly decreased accumulation ofα-syn.The m RNA levels of HMGCR,ABCA1 and A20 in SN were decreased by LPS treatment,which was attenuated by the injection of LV-sponge.CONCLUSION The selective regulation of miR-873 can protect the dopaminergic neurons from the LPS-induced damage.The inhibition of miR-873 can attenuate the relocation of cholesterol in lysosome and the accumulation ofα-syn in neurons induced by LPS via the regulation of HMGCR,ABCA1 and A20.

Risk factors for Parkinson disease and the path analysis: One-to-one paired design

BACKGROUND： Parkinson disease （PD） results from the reduce of neurotransmitter dopamine that transmits intracellular information in brain caused by some reasons, then leads to the dynamic disequilibrium with another neurotransmitter of acetylcholine which is relatively hyperactive. The main causes for PD are still unclear. OBJECTIVE： To screen out the risk factors of PD by means of univariate analysis and multivariate Logistic regression analysis, and investigate the manner of actions between various factors and PD, so as to provide clues for the etiological study of PD. DESIGN： A paired design, Logistic regression analysis, path analysis. SETTING： Department of Scientific Research, Shandong Institute of Physical Education. PARTICIPANTS： Totally 157 PD patients were selected from the Department of Neurology, Qilu Hospital of Shandong University from November 2001 to October 2002. Inclusive criteria： PD was diagnosed according to the standard set by the Fourth National Seminar on Extrapyramidal Disease, Parkinsonian syndromes caused by stroke, carbon monoxide poisoning, encephalitis, drugs, etc. were excluded. Another 157 patients treated in the same department at the same period were selected as the control group, they were the same in sex as those in the patient group, within 3 years older or younger than those in the patient group, and without PD or other extrapyramidal diseases. METHODS： （1） The general conditions were investigated in all the subjects, including general conditions, social behavioral factor, environmental factor, genetic factor, life events, and previous disease; There were 12 main variables, including educational level, family history, mental labour, contact to insecticides, living place before school-age, smoking index, drinking index, tea-drinking index, history of brain trauma, history of cardiovascular disease, history of diabetes mellitus, and history of depression. （2） SAS6.12 software and SPSS 10.0 software were used in the conditional Logistic regression analysis and path analysis respectively. MAIN OUTCOME MEASURES： The results of 12-variable univariate and multivariate analyses; Correlation between main variables and PD; Effects of the factors. RESULTS： All the subjects were involved in the analysis of results. （1） The results of Logistic regression analysis showed that family history, mental labour, insecticides, drinking index and history of depression all had significant positive correlations with PD （univariate analysis： OR=1.405- 5.429, P 〈 0.05- 0.01; multivariate analysis： OR=2.029- 6.754, P 〈 0.05- 0.01）, whereas smoking had significant negative correlations with PD [univariate analysis： odd ratio （OR）=0.765, P 〈 0.05; multivariate analysis： OR =0.489, P 〈 0.01]. （2） The path analysis showed that family history, mental labour, insecticides, smoking, drinking and history of depression had direct effects on PD occurrence [（path coefficient= - 0.218 to 0.204, P 〈 0.05 -0.01）]; Insecticides could cause PD indirectly on the basis of family history （genetic susceptibility） （path coefficient=0.946, P 〈 0.01）; Insecticides could also cause PD by drinking （path coefficient=0.165, P 〈 0.01） Drinking could cause PD indirectly on the basis of family history （path coefficient=0.043, P 〈 0.01 ）. CONCLUSION： The main risk factors of PD are family history, history of depression, drinking, mental labour and insecticides, whereas the protective factor is smoking. PD attack has genetic susceptibility, insecticides and drinking can cause PD on the basis of PD family history. The risk of PD can be decreased by reducing the occasion for contacting the environmental risk factors.

Dopamine transporter distribution in patients with Parkinson disease of different stages detected using single-photon emission computed tomography brain imaging

BACKGROUND： Literatures have reported that the density changes of dopamine transporter is negatively correlated with the severity degree and grading of disease condition of Parkinson disease （PD）. However, the distribution ofdopamine transporter in each nucleus of corpora striatum at each period is still unclear. OBJECTIVE： To observe the radioactive uptake distribution of dopamine transporter in bilateral corpora striata of patients with different stages of PD using single photon emission computed tomography （SPECT）, and make a comparison with healthy controls. DESIGN： Case-control analysis. SETTING： Department of Imageology, Second Hospital Affiliated to Guangzhou University of Chinese Medicine. PARTICIPANTS： Thirty patients with PD admitted to Second Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine between January and December 2005 were recruited. The involved patients, 19 male and 11 female, were aged from 36 to 80 years and with disease course of 2.5 months to 10 years. They all met the clinical diagnosis criteria of Britain Parkinson＇s disease Association Think Tank; Following Hoehn-Yahr grading： grade Ⅰ ： unilateral morbidity; grade Ⅱ： bilateral morbidity, but without balance disorder; grade Ⅲ： bilateral morbidity, accompanied with early posture balance disorder; grade Ⅳ： severe morbidity, needs more help; grade V ： without help, only in bed or wheelchair. There were 11 patients with mild PD （grade Ⅰ - Ⅱ ）, 9 patients with moderate PD （grade Ⅲ） and 10 patients with advanced PD （grade Ⅳ - Ⅴ ）. Meanwhile, 6 healthy persons were selected as normal controls. Informed consents were obtained from all the subjects. METHODS： Twenty-four hours after withdrawal of PD drugs, 30 patients with PD and 6 healthy controls took kalium perchloricum 400 mg orally. After lying down for 30 minutes, all the subjects were intravenously injected with 740 MBq 99Tc m-TRODAT-1 （Jiangsu Institute of Atomic Medicine, Batch No. 20040310） at elbow part. Following injection, image was collected using scanner for single photon emission computed tomography （ADAC Company, USA）. Matrix was 64 × 64, each detecting head revolved 180 ° , 1 frame/60 s. Sixty-four frames were collected with double detecting heads, 50 K/frame. Faultages with clearest image of corpora striatum were selected. Regions of interest （ROI） of caudate nucleus, anterior and posterior putamen and thalamic region in bilateral corpora striata were radioactively counted, and mean value of radioactive counting of ROI was used as the mean value ofpixel in each region of bilateral corpora striata. MAIN OUTCOME MEASURES： Comparison of radioactive uptake in each region of brain between healthy persons and patients with PD. RESULTS： Thirty patients with PD and six healthy persons who received body examination participated in the final result. Comparison of radioactive uptake in each region between healthy persons and patients with PD： ①In the healthy persons, high-density radioactive uptake was found in bilateral corpora striata; Structures of caudate nucleus, anterior and posterior putamen, and thalamus were clear with eudipleural radioactive distribution, and the background of peripheral brain tissue was very low. ②Radioactive intakes in opposite anterior and posterior putamen of patients with mild PD were significantly inferior to those in homolateral ones（70.45±3.35, 87.64±2.65, t =15.82, P 〈 0.05）. Structures of bilateral caudate nucleus and thalamus were clear with eudipleural radioactive distribution （P 〉 0.05）. ③Radioactive intakes in anterior and posterior putamen and thalamus of patients with moderate PD were significantly reduced as compared with healthy persons. There were significant differences in mean radioactive counting of ROI between patients with PD and healthy persons （t =5.20, P 〈 0.05： t =3.95, P 〈 0.05）; The structure of opposite caudate nucleus was not very clear, radioactive distribution of opposite caudate nucleus was significantly reduced as compared with homolateral one （81.11±4.25, 104.56±3.64, t = 14.65, P 〈 0.05）. ④As for patients with advanced PD, the structure of bilateral corpora striata was not clear, radioactive intake was significantly reduced and peripheral background was heightened, even higher than the distribution of the whole corpora striatum. CONCLUSION： SPECT DAT imaging of brain can show the distribution of radioactive uptake in each region of bilateral corpora striata of patients with different stages of PD, which is helpful to diagnose and evaluate the severity of PD.

Loss of microglial autophagy causes Parkinson disease-like symptom in mice

Microglia activation induced neuroinflammation closely relates with the development of Parkinson disease. Autophagy regulates many biological processes,but the role in microglial activation and the development of Parkinson disease is not clear. In this study,we show that loss of microglial Atg5 cause neuroinflammation and motor and cognitive learning impairment in mice,with accumulation of α-synuclein and decrease of dopamine levels in the striatum. Inhibition of autophagy aggravates the activation of NLRP3 inflammasome via PDE10a-cAMP signaling in microglia. Furthermore,the downstream cytokine IL-1 increases Mif levels in a transcriptional dependent manner. Interestingly,Mif levels are significantly elevated in Parkinson disease patients. Taken together,our results reveal a protective role of autophagy in microglial activation-driven Parkinson disease,thus providing a potential targets for the clinical treatment.

Establishing motor disorder mouse models of Parkinson disease Comparison of 6-hydroxydompamine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine

At present, pathogenesis and mechanism of Parkinson disease （PD） are still unclear. Animal models of PD are essential tools in studies on etiology and therapy and should mimic the chronic pathological process, histological characteristics and motor behavior dysfunction. In recent years, transgenic mice have been widely utilized to study the mechanism of PD, and it has become imperative that a PD mouse model of motor behavioral dysfunction be established. OBJECTIVE： To compare the behavioral and histochemical characters of two neurotoxic mice model induced with 6-hydroxydopamine （6-OHDA） or 1-methyl-4-phenyl-1, 2, 3, 6 -tetrahydropyridine （MPTP）, and a better method to mimic Parkinson disease will be found out. DESIGN： Parallel experiment. SETTING： Laboratory of Molecular Genetics, Department of Medical Genetics, Shanghai Jiao Tohg University. MATERIALS： Sixty 129Sv/C57BL6J male wild mice, SPF grade, 8 - 12 weeks old, weighing 20 - 25 g, were provided by Experimental Animal Center, Shanghai Jiao Tong University. All the surgery operation was performed according to the rules of Shanghai Jiaotong University Animal Committee. METHODS： The experiment was carried out in the Laboratory of Molecular Genetics （National Key Laboratory）, Department of Medical Genetics, Shanghai Jiao Ttong University from March to August 2006. ①Thirty-two male mice were randomly divided into control group and drug treatment group with 16 mice in each group. Surgery was carried out and 6-OHDA was administrated to substantia nigra pars compacta （SNpc） and nigra-striatum pathway according to the different parameters with intoxication apparatus. Saline was injected to the other 16 mice according to the same paradigm. 1 mg/kg apomorphine was injected intraperitoneally 2 weeks later after surgery to induce the imbalanced rotation behavior for 40 minutes. ②Twenty-eight mice were randomly divided into 4 groups with 7 in each group, including low-dose, moderate-dose, high-dose groups and negative control group. Then, mice in the drug treatment group were injected intraperitoneally with 5, 10 and 15 mg/kg MPTP for 9 successive days. In addition, mice in the control group were injected with the same volume of saline for 9 days. Pole test and stride length test were utilized to detect coordinative behavioral dysfunction. Mice were sacrificed 20 days after MPTP treatment, and histochemical staining of tyrosine hydroxynase （TH） was used to observe the loss of dopaminergic neuron in SNpc. MAIN OUTCOME MEASURES： ① Success ratio of each model establishment method; ② inducible asymmetric cycle behavior test 2 weeks after 6-OHDA injection; ③behavioral dysfunction in pole test and stride length, morphological changes in brain tissue. RESULTS： Totally sixty mice were used in this experiment and 3 mice were excluded because of the hypersensitivity or the clumsy reaction in motor behavioral detection before MPTP treatment, therefore, data was analyzed with the rest 57 mice. ① Lethal ratio： Three out of 16 mice died in striatum injection group and 5 out of 16 mice died in nigro-striatal pathway group. No mouse died in MPTP treatment groups. ② Locomotor behavior： No dysfunction of locomotor was found in 6-OHDA treatment groups. However, several motor behavioral dysfunction were start to present at the 4th day of MPTP injection. ③ Asymmetric cycle behavior： No asymmetric cycle was induced successfully two weeks after 6-OHDA surgery. Mice show hypersensitive behavior 10 minutes after apomorphine injection, which lasted for about 20 minutes. ④ Pole test： From the 4^th day of MPTP treatment, mice started to display coordinate dysfunction, such as climbing down along the pole in spiral, moving slowly with hesitation. Some mice could not grab the pole and slide down along the pole at 4th day post injection. Comparing with 0 dose control group, all the threedrug treatment groups show significant different dysfunction from the 4th day to the 20th day post injection （P 〈 0.01）. ⑤ stride length test： Mice＇s stride length decreased, when treated with MPTP, and the mice in the high dose group displayed obviously. ⑥ Dopaminergic neuron stained with TH in nigra pars compacta： The results indicated that administrated MPTP （from low dose to high dose） by intraperitoneal cause chronic lesions on the dopaminergic neuron in the SNpc. CONCLUSION： PD mice models induced with 6-OHDA show high mortality ratio and no asymmetric cycle was found after apomorphine injection. However, injection of MPTP intraperitoneally can simulate the chronic pathway of PD, typical histological changes are found and stable motor behavioral dysfunctions are displayed.

Research progress in mechanism of curcumin in treatment of Parkinson disease

Curcumin(Cur)is an important bioactive component of polyphenols in the rhizomes of Curcuma longa L.,Tulipa gesneriana L.and other Curcuma plants.It has a wide range of pharmacological effects such as anti-tumor,anti-atherosclerosis,anti-inflammatory,and neuroprotection.Parkinson disease(PD)is a neurodegenerative disease that often occurs in the elderly.Its main pathological characteristics are the characteristic loss of substantia nigra dopaminergic neurons,the decrease of dopamine content in the striatum,and the formation of Lewy bodies.At present,the main methods of clinical treatment of PD include drug therapy and surgical operation,but due to its complicated pathogenesis,they can only play a role in relieving,but cannot be completely cured.Modern pharmacological studies have shown that Cur has certain effects in the treatment of PD.①Anti-oxidative stress:oxidative stress is closely related to the degeneration of dopaminergic neurons.Studies have found that Cur can increase the activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),reduce malondialdehyde(MDA)content,thereby reducing oxidative stress damage and protecting dopaminergic neuron.②Reduce inflammation in brain tissue:neuroinflammation plays an important role in the development of PD.Reducing the level of inflammatory factors can have a certain therapeutic effect on PD.Studies have shown that high-dose Cur can reduce the levels of interleukin-6(IL-6),IL^(-1)β,and tumor necrosis factor-α(TNF-α)in brain tissue,reduce inflammation,inhibit further neuronal damage,improve learning and memory,and exert neuroprotective effects.③Activation of autophagy:the abnormal accumulation ofα-Synuclein(α-Syn)in Lewy bodies is closely related to PD,and autophagy dysfunction leads toα-Syn clearance obstacles and an important factor of abnormal aggregation.Cur can increase the expression of microtubule-associated protein 1 light chain 3(LC3-Ⅱ)and lysosome-associated membrane protein 2A(LAMP2A),and reduce the protein and mRNA expression ofα-Syn.It can be seen that Cur promotes the elimination ofα-Syn and protects neurons from damage by activating autophagy.④Inhibition of mitochondrial dysfunction:mitochondria plays a central regulatory role in the process of cell apoptosis,and mitochondrial dysfunction is related to reactive oxygen species,energy and mitochondrial membrane potential,which may cause substantia nigra striatal neuropathy.Experiments have shown that Cur can reduce the active oxygen content in PC12 cells induced by MPP+,maintain the normal membrane potential of mitochondria,thereby stabilizing mitochondrial function and inhibiting PC12 cell apoptosis.This study summarized the action mechanism of Cur in the treatment of PD,and clarified the basis of its pharmacodynamics, providing a reference for the clinical research and new drug develop ment research of Cur in the treatment of PD.

Urolithin A protects dopaminergic neurons in experimental models of Parkinson disease by promoting mitochondrial biogenesis through SIRT1/PGC-1αsignaling pathway

OBJECTIVE Mitochondrial dys⁃function contributes to the pathogenesis of neuro⁃degenerative diseases such as Parkinson dis⁃ease(PD).Therapeutic strategies targeting mito⁃chondrial dysfunction hold considerable promise for the treatment of PD.Urolithin A(UA)is a gut metabolite produced from ellagic acid-containing foods such as pomegranates,berries,and wal⁃nuts.Recent reports have highlighted the protec⁃tive role of UA in several neurological disorders including Alzheimer disease and ischemic stroke.However,the potential role of UA in PD has not been characterized.In this study,the role of UA in 6-OHDA-induced neurotoxicity in cell cultures and mouse model of PD was investi⁃gated.METHODS In vitro,PC12 cells were exposed to 6-OHDA in the presence or absence of UA.For in vivo study,C57BL/6 mice were ste⁃reotactic injected with 6-OHDA to induce experi⁃mental PD model.UA(10 mg·kg-1)was intraperi⁃toneal injected for 7 d before surgery.RESULTS UA protected against 6-OHDA cytotoxicity and apoptosis in PC12 cells.Prior administration of UA to 6-OHDA lesioned mice ameliorated both motor deficits and nigral-straital dopaminergic neurotoxicity.Moreover,UA attenuated 6-OHDA-induced mitochondrial dysfunction in PC12 cells accompanied by enhanced mitochondrial biogen⁃esis.Mechanically,the neuroprotective effects of UA were mediated by SIRT1-PGC-1αsignaling-mediated mitochondrial biogenesis.CONCLU⁃SION These data provide new insights into the novel role of UA in promoting mitochondria bio⁃genesis and suggest that UA may have potential therapeutic applications for PD.