Objective To observe the relation between onset age and the motor complications involved Parkinaon' s disease (PD).Methods The detailed clinical information of 195 patients with idiopathic PD and good response to L-dopa were recorded and followed up.The data were calculated with SPSS statistic software.Results Although the time interval between the onset of the disease and the use of L-dopa was significantly longer in the 57 early-onset patients as compared to the 138 later-onset ones ((3.88±3.33) years vs (2.36±2.21) years, t = 3.142, P = 0.002), the time interval between the use of L-dopa and the occurrence of motor complications was not significantly longer in the early-onset group ((3.81±2.06) years vs (4.24±2.00) years, t = -0.888, P = 0.378).There was no difference in the constituent ratio of non-L-dopa use from onset between early-onset and later-onset PD groups (28.07% (16/57) vs 27.54% (38/138), χ2 = 0.006, P = 0.940).There was also no difference in the dosage of daily L-dopa use when motor complications occurred between early-onset and later-onset PD groups ((601.8± 296.7) mgvs (655.6±192.5) mg, t = -0.912, P=0.365).Seven-tenths of the patients with an onset age younger than 40 years old carried the risk of motor complications after using L-dopa for 5-years and those older than 70 years had the risk at a rate of 1/10.Conclusions Delaying the of use of L-dopa may not necessarily delay the onset of motor complications.The high incidence of motor complications among younger patients may not be related with drug dosage and the type of drug firstly chosen.Younger onset age does inerease the ineidence of motor complicatious.

OBJECTIVETo establish an allele-specific PCR method for detect screening of CYP21A2 gene mutation.

METHODSAllele-specific PCR primers and analogy primers were designed based on the sequence alignment of CYP21A2 and CYP21AP genes. Genomic DNA was extracted from blood specimens of 4 patients with 21-hydroxylase deficiency and 5 healthy controls and respectively amplified with allele-specific PCR primers and analogy primers and sequenced.

RESULTSMutations of CYP21A2 including IVS2-13A/C>G, Arg356Trp and Arg149Pro were found with the established method in all of the 4 patients but not in the healthy controls. When detected with the analogy primers set, IVS2-13A/C>G and Arg356Trp were observed in both patients and healthy controls.

CONCLUSIONThe allele-specific PCR-based method is a simple, effective and reliable method for the detection of CYP21A2 gene mutation.

Adrenal Hyperplasia, Congenital ; enzymology ; genetics ; Alleles ; Base Sequence ; DNA Mutational Analysis ; methods ; DNA Primers ; genetics ; Humans ; Molecular Sequence Data ; Mutation ; Polymerase Chain Reaction ; methods ; Steroid 21-Hydroxylase ; genetics

Objective To investigate the main cell types expressed brain-derived neurotrophic factor (BDNF) in the posterior cortical infarction area in cerebral infarction rats after early vein allograft of bone marrow mesenchymal stem cells (BM-MSCs) and the effect of BM-MSCs transplantation on their ce11 numbers and percentages.Methods (1) Fifteen SD rats were randomly divided into sham-operated group 1,ischemia control group 1,and BM-MSCs transplantation group 1 (n=5);distal middle cerebral artery occlusion (dMCA) models were used in the later two groups;1 × 106 CM-DiI labeled BM-MSCs were intravascularly transplanted into the tail vein of rats from the transplantation group 1 at one h after ischemia;all rats were sacrificed 48 h after ischemia;BM-MSCs with co-existence of CM-Dil and BDNF in the ischemia cortex areas were detected by immunofluorescence staining.(2)Fifteen SD rats were randomly divided into sham-operated group 2,ischemia control group 2,and BM-MSCs transplantation group 2 (n=5);dMCAO models were used in the later two groups;1 ×106 non-labeled BM-MSCs were intravascularly transplanted into the tail vein of rats from the transplantation group 2 at one h after ischemia;48 h after ischemia onset,all rats were sacrificed;the number of BDNF+ and CD68+ microglia cells,BDNF+ and Iba-1+ microglia cells,and BDNF+ and neuron-specific nucleoprotein (NeuN)+ neurons were measured by immunofluorescence staining.Results (1) CM-Dil red fluorescence labeled allogeneic BM-MSCs were only found in BM-MSCs transplantation group 1;the labeled cells scattered in the infarct and peri-infarct cortices;9.70％±3.47％ CM-Dil labeled BM-MSCs expressed BDNF,accounting for 13.32％±4.48％ of all BDNF+ cells in the infarct brain cortex.(2) In the brain tissues of cortex infarct area of BM-MSCs transplantation group 2,38.40％±9.04％ BDNF+ cells were Iba-1+ microglia cells,11.65％±2.76％ BDNF+ cells were CD68+ microglia cells,and 28.96％±6.99％ BDNF+ cells were NeuN+ neurons;the Iba-1+ cell numbers and Iba-1+/BDNF+ double positive cell percentages in the BM-MSCs transplantation group 2 ([92.06±36.52]/mm2 and 79.21％±12.27％) were significantly increased as compared with those in the ischemia control group 2 ([31.13±10.23] mm2 and 60.15％±28.20％,P＜0.05).Conclusion Allogeneic BM-MSCs is capable of migrating into the infarct cortex when intravenous transplantation of BM-MSCs is performed at the early stage after ischemia;the main sources of BDNF in these areas are microglias cells and neurons;these BM-MSCs increase both number and percentage of Iba-1+/BDNF+ double positive cells,which may be one of the underlying mechanisms of therapeutic effects.

In Parkinson's disease (PD),long-term use oflevodopa-induced dyskinesia (LID) significantly affects the therapeutic efficacy of levodopa and quality of life of patients.Current existing therapies are not ideal,which is related to that mechanism of LID has not been fully clarified.Recently,more and more studies have confirmed that the glutamatergic system is closely related to the dopaminergic system,and the role of glutamate receptors in LID is increasingly prominent,especially,metabolic glutamate receptor (mGluR)4,mGluR 5 and partial ionotropic glutamate receptors are current research hotspots in the mechanism and clinical observation of drugs of LID.Therefore,this review summarizes the changes and roles of various glutamate receptors in LID and related clinical studies,hoping to provide new ideas for the diagnosis and treatment of LID in PD patients.

OBJECTIVETo identify potential mutation of the colony stimulating factor 1 receptor gene (CSF1R) in a large Chinese family affected with hereditary diffuse leukoencephalopathy with spheroids (HDLS) and analyze the genotype-phenotype correlation.

METHODSThe proband was evaluated physically and radiologically to ascertain the HDLS phenotype. Genomic DNA was extracted from peripheral blood samples from family members. The coding region of the CSF1R gene was amplified with PCR and subjected to direct DNA sequencing.

RESULTSThere were 9 affected members (5 alive) in this five-generation family (1 member had died during the follow-up). A missense mutation c.2563C>A (p.P855T) of the CSF1R gene has been identified in the proband. The same mutation was identified in 3 affected and 1 unaffected members of the family.

CONCLUSIONThe family was consistent with autosomal dominant inheritance. CSF1R gene mutation is also a disease-causing mutation in Chinese patients.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Female ; Genes, Dominant ; Humans ; Leukoencephalopathies ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Pedigree ; Receptor, Macrophage Colony-Stimulating Factor ; genetics

Objective To investigate whether human bone marrow mesenchymal stem cells (hBC-MSCs) have the immune-suppression ability on the proliferation of peripheral blood mononuclear cells (PBMCs) and their pro-inflammatory cytokine production in rats,to explore what kinds of human cytokines are required for the induction of hBM-MSCs to become immune-suppressive,and to observe the effect of intravenous delivery of hBM-MSCs on tumor necrosis factor (TNF)-α transcription and expression in the core infarct areas of rats after cerebral ischemia.Methods The fetal-originated hBC-MSCs and rat PBMCs were extracted;the rat PBMCs were activated by adding 10 μg/mL concanavalin A (ConA).(1) The first experiment was divided into hBM-MSCs+PBMCs group,hBM-MSCs+PBMCs+ConA group,PBMCs group and hBM-MSCs group;CCK-8 assay was employed to detect the proliferation of these cells.(2) The second experiment was divided into hBM-MSCs+PBMCs+ConA group,PBMCs+ConA group;ELISA was used to detect the TNF-α,interferon-γ (IFN-γ) and intedeukin (IL)-10 expressions.(3) The third experiment was divided into hBM-MSCs+PBMCs (IFN-γ+IL-1α) group,hBM-MSCs+PBMCs (IFN-γ+IL-1β) group,hBM-MSCs+PBMCs (IFN-γ+TNF-α) group and hBM-MSCs+PBMCs group;CCK-8 assay was used to detect the proliferation of these cells.(4) Thirty SD rats were randomly divided sham-operated group,control group (giving normal saline after ischemia) and hBM-MSCs group (giving hBM-MSCs after ischemia,n=10);on the third d of ischemia,the TNF-α mRNA and protein expressions at the infarct areas was detected by real time PCR and Western blotting,respectively.Results (1) The optical density (OD) in the hBM-MSCs+PBMCs group was significantly increased as compared with that in the PBMCs group and hBM-MSCs group (P＜0.05);OD in the hBM-MSCs+PBMCs group was significantly increased as compared with that in the hBM-MSCs+PBMCs+ConA group (P＜0.05).(2) The TNF-α,IFN-γand IL-10 levels in the PBMCs+ConA group were (1030±196) pg/mL,(2880±250) pg/mL and (330±45) pg/mL;the TNF-α and IFN-γlevels in hBM-MSCs+PBMCs+ConA group were (160±10) pg/mL and (240±55) pg/mL,which were significantly lower than those in the PBMCs+ConA group (P＜0.05);the IL-10 level in hBM-MSCs+PBMCs+ConA group was (750±110) pg/mL,which was significantly higher than that in the PBMCs+ConA group (P＜0.05).(3) The OD in the hBM-MSCs+PBMCs(IFN-γ+IL-1α)group,hBM-MSCs+PBMCs (IFN-γ+IL-1 β) group and hBM-MSCs+PBMCs (IFN-γ+TNF-α) group was significantly decreased as compared with that in the hBM-MSCs+PBMCs group (P＜0.05).(4) The TNF-α mRNA expression in the sham-operated group,control group and hBM-MSCs group was 0.490±0.128,2.369±0.788 and 1.002±0.408;the TNF-α protein expression in the sham-operated group,control group and hBM-MSCs group was 0.144±0.028,0.314±0.029,0.240±0.029;the TNF-α protein and mRNA expressions in the hBM-MSCs group were significantly decreased as compared with those in the control group (P＜0.05).Conclusions The allogeneic transplantation of hBC-MSCs is competent in suppressing the inflammation of rats in vitro and in vivo.Furthermore,this immune-suppression ability is not innate,but cytokine stimulation dependent.The immune-suppression ability of hBM-MSCs on rat PBMCs are at least partly responsible for the therapeutic effect of hBM-MSCs transplantation into the rat models,such as ischemia stroke.

Objective:To construct an automatic recognition system for PD patients with freezing of gait (FOG) based on mobile phone videos by recording the gait videos of PD patients with FOG.Methods:Forty-nine PD patients with FOG, admitted to our hospital from December 2020 to May 2021, were chosen in our study. Their clinical data were collected. The processes of these patients accepted "3-meter-round trip" and "3-meter-round trip through narrow (0.6 m)" were recorded and 87 valid gait videos were extracted. Position signals of key points in the video were extracted, and featured data were extracted after signal preprocessing. From the featured data, action recognition model, straight FOG recognition model and turn FOG recognition model were established respectively, and finally end-to-end FOG recognition model was formed. Leave-one-subject-out (LOSO) method was used to evaluate the performance of the above models.Results:A total of 22 066 non-FOG window samples and 3815 FOG window samples were obtained from 87 valid videos, which constituted the training sample pool of this study. LOSO method showed that the motion recognition model enjoyed 83.27% sensitivity, 91.38% specificity, and 89.28% accuracy; the straight FOG recognition model enjoyed 57.69% sensitivity and 88.12% specificity; the turn FOG recognition model enjoyed 61.54% sensitivity and specificity 98.72%; and the end-to-end FOG recognition model enjoyed 85.71% sensitivity and 75.73% specificity.Conclusion:The automatic recognition system for PD patients with FOG based on mobile phone videos has relatively high sensitivity and specificity, which can realize remote assessment and is convenient for screening and follow-up of PD patients with FOG.

Objective To evaluate the diagnostic value of lymph node fine-needle aspiration (FNA)Epstein-Barr virus (EBV)-DNA concentration detection in nasopharyngeal carcinoma (NPC) cervical lymph node metastasis.Methods From August to December 2016,36 cases of NPC and 9 cases of other tumors (not correlated with EBV infection) were enrolled in this study at the Sun Yat-sen University Cancer Center.All patients received magnetic resonance images (MRI),plasma and cervical lymph node FNA EBV-DNA detection.Results The median concentration of EBV-DNA in FNA fluid (1.39 × 105 copies/ml) in cervical lymph node metastasis was significantly higher than that in plasma (2.00 × 103 copies/ml),with a significant difference (x2 =16.723,P =0.004).The diagnosis sensitivity,specificity,accuracy of the lymph node FNA fluid of EBV-DNA were 86.2％ (25/29),71.4％ (10/14) and 81.4％ (35/43) respectively,which were better than those of MRI [72.4％ (21/29),50.0％ (7/14) and 65.1％ (28/43) respectively] and plasma EBV-DNA [55.2％ (16/29),71.4％ (10/14) and 60.5％ (26/43) respectively].The area under the curve (AUC) of level Ⅰ b cervical lymph node metastasis was calculated,and FNA fluid EBV-DNA (AUC =0.688)was better than MRI (AUC =0.583),with a significant difference (Z =2.476,P =0.008).The EBV-DNA concentration in FNA fluid in cervical lymph node metastasis of patients with other tumors (no correlated with EBV infection) was 0 copy/ml.Conclusion FNA fluid EBV-DNA may improve the diagnostic sensitivity of cervical lymph node metastasis in nasopharyngeal carcinoma,and help to explore the clinical target volume neck nodes at level Ⅰ b cervical lymph node in radiotherapy.