Advances in genetics research in the pathogenesis of amyotrophic lateral sclerosis / 中南大学学报(医学版)

Amyotrophic lateral sclerosis (ALS) is a rare neurodegenerative disease affecting the upper and lower motor neurons. It is characterized by progressive muscle weakness, atrophy and ultimate death due to dysphagia and dyspnea. There are many causes of ALS, among which the genetic factors show great relevance. Imbalance of protein homeostasis in neurons, prion-like proliferation and propagation of abnormal proteins, mitochondrial dysfunction, glutamate mediated excitotoxicity, and intraneuronal substance transport disorders are recognized as the pathogenesis.The study on gene mutation related to pathogenesis will bridge the molecular and cellular research of ALS, which can deepen the understanding of the occurrence and development of ALS and the role of gene mutation in ALS, and provide new ideas and enlightenment for the treatment of ALS.

Proteomic analysis of the cerebrospinal fluid from patients with amyotrophic lateral sclerosis based on tandem mass spectrometry technique / 南方医科大学学报

OBJECTIVE@#To investigate the differentially expressed proteins in the cerebrospinal fluid (CSF) of patients with amyotrophic lateral sclerosis (ALS) at the proteomics level using tandem mass spectrometry label (TMT) technique and explore the pathogenic mechanism and related pathways of ALS.@*METHODS@#Between November, 2017 and April, 2018, 5 patients with medulla oblongata onset ALS and 5 patients with limb onset ALS were selected from the Departments of Neurology of 928 Hospital of Army Joint Logistics Support Force of PLA and Xiangya Hospital of Central South University, with 5 patients with migraine and low intracranial pressure headache serving as the healthy controls.CSF samples were obtained from all the participants, and the differentially expressed proteins in the CSF were identified using tandem mass spectrometry (TMT) technique with bioinformatics analysis.@*RESULTS@#A total of 1530 proteins were identified and quantified in the CSF samples.The expression of 48 proteins was up-regulated and 6 proteins were down-regulated in medulla oblongata onset ALS patients; 16 proteins were up-regulated and 19 were down-regulated in limb onset ALS patients.GO analysis showed that these proteins, which were distributed both within and outside the cells, were involved in cell physiological process, single organ process and biological regulation and had binding function, catalytic activity, and receptor activity.KEGG pathway analysis showed that the up-regulated proteins in the CSF from patients with medulla oblongata onset ALS participated in 3 pathways involving the lysosomes, metabolism, and measles.The down-regulated proteins in the CSF from patients with limb onset ALS participated in 7 pathways involving the complement and coagulation cascade, infection and herpes simplex infection, and all the pathways contained complement components.@*CONCLUSIONS@#The CSF samples of ALS patients with medullary onset and limb onset have differentially expressed proteins.The lysosomal pathway is involved in the occurrence and progression of ALS with medullary onset, and the immune responses are involved in the occurrence and progression of ALS with limb onset.

Clinical features of anti-N-methyl-D-aspartate receptor encephalitis and the concomitant seizure / 中南大学学报(医学版)

To investigate the clinical features, auxiliary examination and characteristics for anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis and its concomitant seizure.
 Methods: A total of 20 patients diagnosed as anti-NMDAR encephalitis were enrolled from January 2016 to September 2018 in Xiangya Hospital. The data including the clinical features, auxiliary examination, characteristics of seizure, treatment and prognosis were collected. The discharged patients were followed up for half a year.
 Results: The initial symptom in patients with anti-NMDAR encephalitis were mainly psychiatric symptom and seizure. Most of the EEG result were diffused slow waves. The mainly type of seizure in patients with anti-NMDAR encephalitis showed generalized tonic-clonic seizure. Patients occurred consciousness during the onset of the disease. MRI showed that patients with temporal lobe were more inclined to occur seizure than patients with anti-NMDAR encephalitis (P<0.05). After standardized treatment, 20 patients showed a significant improvement in modified Rankin Scale (mRS) scores and the seizure was under control within half a year. 
 Conclusion: Patients with temporal lobe affected in MRI should pay attention to the possibility of seizure occurrence. Anti-epileptic drugs and immunotherapy should be used promptly in patient with seizure. After standardized treatment, the prognosis of patients will be mostly good.

Risk factors of death in 30 days after syncope / 中华急诊医学杂志

Objective To investigate the risk factors of death in patients with syncope. Methods Clinical data of 516 patients experienced syncope admitted from June 2010 to June 2016 were analyzed retrospectively. Factors including gender, age, history of hypertension, diabetes mellitus, hyperlipidemia, smoking history, drinking history, and etiology of syncope (cardiogenic syncope, neuroreflex syncope, orthostatic hypotension, orthostatic syncope, unexplained syncope, and syncope caused by other special diseases) were analyzed as likely risk factors of death within 30 days after syncope happened. After adding the derived variables (over 22 new factors), analyses were done to investigate independent risk factors of death for patients with syncope. Results This study included 321 male (62.2％) and 195 females (37.8％), with mean age of (62.23±19.69) years. Logistic regression analyses showed that age (OR=1.033, 95％ confidence interval (95％CI):1.008-1.058, P =0.008 8),cardiac syncope (OR=19.704,95％CI:5.894-5.875,P＜0.01) were independent risk factors of death within 30 days after syncope occurred. Multiple-variate analysis with derived variables showed that cardiac syncope (OR=11.487, 95％CI:4.938-26.721,P＜0.01),age and age derived variables (OR=1.000, 95％CI:1.000-1.000,P=0.000 8),age and cardiogenic syncope derivative variables (OR=1.033, 95％CI:1.022-1.044, P＜0.01) were independent risk factors for death within 30 days after syncope. Conclusion Age and cardiogenic syncope were independent risk factors for death within 30 days after syncope occurred. And a derivative factor of age, and interactivity between age and cardiac syncope were independent risk factors of death in patients with syncope.

Mutations of G38R and D40G cause amyotrophic lateral sclerosis by reducing Annexin A11 protein stability / 中南大学学报(医学版)

To explore the role of the mutations G38R and D40G of Annexin A11 (ANXA11) in the onset of amyotrophic lateral sclerosis (ALS).
 Methods: The plasmids expressing ANXA11 wild type protein, ANXA11 G38R protein and ANXA11 D40G protein were constructed, respectively. The recombinant plasmids were then transfected into HEK293 cells respectively followed by cycloheximide (CHX) treatment for 0, 2, 4 and 8 h. The protein expressions of ANXA11 wild type, ANXA11 G38R and ANXA11 D40G mutations were determined by Western blot. Gray analysis by Image J was performed to compare the half-life of each protein. The NSC-34 cell lines constantly expressing ANXA11 wild type protein, ANXA11 G38R protein and ANXA11 D40G protein were established. The cells were treated with NP-40 lysis buffer to examine the protein solubility by Western blot.
 Results: Both ANXA11 G38R protein and ANXA11 D40G protein showed a shorter half-life than ANXA11 wild type protein (P0.05). There was no visible insoluble substance in the NP-40 lysates for ANXA11 wild type protein, ANXA11 G38R protein and ANXA11 D40G protein.
 Conclusion: G38R and D40G mutations reduce the stability of ANXA11 protein. G38R and D40G mutations do not alter ANXA11 solubility.

Clinical features of a genetically identified spinal and bulbar muscular atrophy pedigree / 中南大学学报(医学版)

Spinal and bulbar muscular atrophy (SBMA) is a rare X-linked motor neuron disease with significant phenotypic viability. Here, we present a genetically identified SBMA family without bulbar paralysis or androgen insensitivity. All four male patients presented with progressive lower motor neuron paralysis in all limbs, with distal extremities more dominant. None of them had bulbar palsy or androgen insensitivity. A consistently mild elevated blood creatine phosphokinase (CPK) levels were detected in all patients and the EMG showed a chronic neurogenic damage. Muscle biopsy of propositus indicated a typical neurogenic amyotrophy. Genetic testing for SMA of mutation in SMN1 was negative, while for SBMA of androgen receptor showed the increased CAG repeat in exon 1, suggesting that although bulbar symptoms and androgen insensitivity are characteristic symptoms of SBMA, they are not obligatory for the diagnosis. In adult males with a chronic motor neuron syndrome without upper motor neuron signs, even in absence of the classical features of androgen insensitivity or bulbar findings, genetic testing for SBMA should be strongly considered.

Risk factors analysis of sudden death in patients suspected with pulmonary thromboembolism in emergency room / 中华危重病急救医学

Objective To explore the correlative factors of sudden death in patients suspected with pulmonary thromboembolism (PTE) in emergency room (ER).Methods A retrospective analysis was conducted.The clinical data of 12 patients with sudden death suspected with PTE (sudden death group) in ER of the Air Force General Hospital from January 2011 to June 2014 were analyzed.The non-sudden death group included 35 patients during the same time period who were diagnosed with PTE based on findings of CT pulmonary arteriography (CTPA) and showed no sudden death in ER.Factors,including sex,age,previous operation,tumor,syncope,dyspnea,bilateral or unilateral edema of lower extremity,heart rate (HR),white blood cell count (WBC),D-dimer,arterial partial pressure of oxygen (PaO2),arterial partial pressure of carbon dioxide (PaCO2) and typical clinical manifestation of electrocardiogram (SⅠTⅢQⅢ),were compared between the two groups.The potential predictors of sudden death of PTE were analyzed by logistic regression analysis.Results Young age (years old:51.3±15.5 vs.62.3±14.4),lower PaO2 [mmHg (1 mmHg =0.133 kPa):49.9± 12.3 vs.62.7± 10.2],higher HR (bpm:122.0± 19.5 vs.89.1 ± 18.5) and higher WBC (× 109/L:13.8 ± 6.9 vs.7.2 ± 2.5) were found in sudden death group as compared with those in non-sudden death group (P ＜ 0.05 or P ＜ 0.01).There was no significant differences in D-dimer level and PaCO2 between sudden death group and non-sudden death group [D-dimer (pg/L):986 (891,3 230) vs.2089 (598,3 397),PaCO2 (mmHg):33.0 (28.6,43.4)vs.36.5 (32.9,41.0),both P ＞ 0.05].The syncope,antineoplaston treatment or tumor metastasis within 6 months,operation in previous 4 months,bilateral asymmetrical edema in sudden death group were more than those of the non-sudden death group,and chest pain was less (P ＜ 0.05 or P ＜ 0.01).Difference in gender,dyspnea and typical SⅠTⅢQⅢ in electrocardiogram were not significant between the two groups (all P ＞ 0.05).It was shown by multiple logistic regression analysis that higher HR [odds ratio (OR) =1.124,95％ confidence interval (95％CI) =1.024-1.235,P =0.014] and higher WBC (OR =1.347,95％CI =1.043-1.738,P =0.022) were identified as independent risk factors of sudden death for PTE.Conclusions Gender,dyspnea,typical S Ⅰ TⅢQⅢ in electrocardiogram,PaCO2 and D-dimer seem unrelated to sudden death of patients with PTE.Young age,chest pain,syncope,bilateral asymmetrical edema,antineoplaston treatment or tumor metastasis within 6 months,operation in previous 4 months and low PaO2 were potential predictors of sudden death according to the univariate analysis.Higher WBC and higher HR are independent risk factors of sudden death for PTE patients.

Expression and Epigenetic Regulation of BRCA1 in Chemosensitive and Chemoresistant Ovarian Cancer / 中国医科大学学报

Objective To investigate the expression of BRCA1 in chemosensitive and chemoresistant ovarian cancer specimens,so as to provide a novel insight into the epigenetic mechanism involved in BRCA1 transcription. Methods Serous ovarian cancer patients(10 chemosensitive and 10 chemoresistant cancer)were enrolled for the study. BRCA1 levels was analyzed by real?time quantitative PCR. The methylation levels of BRCA1 core promoter(sites 1?4)was determined by pyrosequencing. Regression analysis was used to examine the possible relationship between BRCA1 levels and the methylation levels of sites 1?4 in ovarian cancer specimens. Results Compared to chemosensitive ovarian cancer tissues,BRCA1 levels were increased,but the methylation levels of BRCA1 core promoter(sites 1?4)were decreased in chemoresistant ovarian cancer tissues. How?ever,it is interesting to note that only a significant inverse correlation was observed between BRCA1 levels and the methylated levels of site 4 (r=-0.612,P<0.05). Conclusion Our findings imply that the methylation levels of site 4 in the core promoter of BRCA1 may be widely involved in the regulation of BRCA1 expression and chemosensitivity in ovarian cancer.

Changes of protein kinase-like endoplasmic reficulum kinase and glucose-regulated protein 78 expression in rats after focal ischemic preconditioning / 中华神经科杂志

ObjectiveTo investigate the effect of focal ischemic preconditioning (IPC) on the expression of protein kinase-like endoplasmic reticulum kinase ( PERK ) and glucose-regulated protein 78 (GRP78) mRNA and protein after focal cerebral ischemia/reperfusion (I/R) in rats.MethodsAll 120 male SD rats were randomly divided into three groups: sham-operation group,middle cerebral artery occlusion (MCAO) group and brain ischemia preconditioning (BIP) group.Each group was further divided into 4 subgroups according to 12 h,1,2 and 3 d after I/R.The IPC models were made in order to measure the expression of PERK,GRP78 mRNA and protein by in situ hybridization and Western blot,and the apoptosis rate of neuron by flow cytometry. Results ①The expression of PERK mRNA increased and reached the peak at 12 h,then decreased continuously after 1 d.BIP could decrease its expression.The expression of PERK protein increased at 12 h and reached the peak at 24 h,then decreased continuously after 2 d.BIP could decrease its expression.②The expression both of GRP78 mRNA and its protein all increased and reached the peak at 12 h,then decreased continuously.BIP could increase their expression (mRNA:12 h: 136.70±9.53,F=32.265; 24 h:147.54 ±9.97,F=54.920; 2 d:158.16 ±9.44,F=45.374; 3d: 165.85±10.26,F=16.493,P＜0.05; protein:12 h: 1.319±0.116,F=5.619,P＜0.05; 24 h: 1.226±0.108,F=33.742,P＜0.01; 2 d:1.183 ±0.112,F =46.556,P ＜0.01; 3 d:1.115± 0.098,F =11.730,P＜0.05).③The rate of apoptosis neuron of rats in MCAO increased markedly at 12 h after reperfusion,and reached the peak at 1 d,then decreased continuously.BIP could decrease the rate of apoptosis neuron. Conclusion BIP can protect neurons through inhibiting the expression of PERK and inducing the expression of GRP78 after endoplasmic reticulum stress in rats.

The expression of XIAP, Smac, HtrA2 and XAF1 in the rat hippocampus following status epilepticus / 中华神经科杂志

Objective To investigate the expression of XIAP, Smac, HtrA2 and XAF1 in the hippocampus following SE in rats and to explore the pathophysiological mechanisms of expression of XIAP and its negative regulators after SE. Methods The lithium-pilocapine model of status epilepticus was established in SD rat. XIAP, Smac, HtrA2, XAF1 and activated caspase-3 protein were examined using immunohistochemistry. Western blot was used to detect the protein levels of XIAP, Smac, HtrA2 and activated easpase-3. Results XIAP immunoreactivity diffusely distributed within the neuron after SE. Compared with the control group, the expression of CA3 XIAP protein in the SE group was increased gradually since 2 hours (0.5503±0.0172 vs 0.1507±0.0165, t=115.87, P<0.01), peaking at 8 hours (0.6221±0.0238 vs 0.1507±0.0165, t=136.69, P<0.01). The expression of CA3 Smac, HtrA2, XAF1 and activated caspase-3 protein were increased generally following SE. Western blot analysis showed a significant increase in Stoat, HtrA2, activated caspase-3 protein levels from 2 to 72 hours following SE, but no significant differences were seen in XIAP protein levels between the control group and the SE group. Conclusions The XIAP, Smac, HtrA2 and XAF1 are involved in the regulation of neuronal apoptosis and implicated in pathophysiological mechanisms of neuronal damage after SE.

Exparimental study on relationship between GAT-1 and neuron injury in focal cerebral ischemia/infusion model / 临床神经病学杂志

Objective To study the relationship between GAT-1 and neuron injury in focal cerebral ischemia/infusion model.Methods Focal cerebral ischemia/reperfusion models were established in rats by thread embolic, then the brain tissues were taken at 3 h, 6 h,12 h, 24 h and 3 d after cerebral ischemia/reperfusion.The number of GAT-1 positive neurons were calculated and the average optical density were detected by immunohistochemical technique in different time points after cerebral ischemia/reperfusion, compared with the normal group and the sham operated group.Results The expression of GAT-1 was up-regulation obviously at 3 h after cerebral ischemia/reperfusion( P