OBJECTIVE To investigate the effects and potential mechanism of persimmon leaf （PL） extract against ferroptosis induced by H2O2 in IEC-6 cells. METHODS Using IEC-6 cells as object， the effects of ferroptosis inhibitor ferrostatin-1 on IEC-6 cell viability induced by H2O2 were investigated； IEC-6 cells were divided into control group， H2O2 group， H2O2+PL 25 μg/mL group and H2O2+PL 50 μg/mL group. The levels of oxidant stress indexes [content of malondialdehyde （MDA）， activity of superoxide dismutase （SOD）， and levels of reactive oxygen species （ROS）]， mitochondrial membrane potential （MMP） as well as mRNA and protein expressions of nuclear factor-erythroid-2 related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， NADPH/quinone oxidoreductase-1 （NQO-1）， cystine/glutamate anti-porter （xCT）， glutathione peroxidase 4 （GPX4） and ferritin heavy chain （FTH） were detected. RESULTS Ferroptosis inhibitor ferrostatin-1 could significantly increase the survival rate of H2O2-induced cells （P＜ 0.01）. Compared with the control group， MDA content， ROS level， mRNA expressions of Nrf2 and NQO-1 as well as protein expressions of Nrf2 and HO-1 were increased or up-regulated significantly， while SOD activity， MMP， mRNA expressions of xCT， GPX4 and FTH as well as protein expressions of GPX4 and FTH were decreased or down-regulated significantly （P＜0.01 or P＜0.05）. Compared with the H2O2 group， oxidative stress Δ indexes of H2O2+PL 25， 50 μg/mL groups were reversed to different extents， MMP level was increased significantly， as well as mRNA and protein expressions of Nrf2， HO-1， NQO-1，xCT， GPX4 and FTH were up-regulated to different extents；there were statistical significances in some indexes between groups （P＜0.01 or P＜0.05）. CONCLUSIONS PL extract can alleviate mitochondrial membrane damage and abnormal accumulation of ROS caused by H2O2， which may be related to the inhibition of ferroptosis by activating the Nrf2/HO-1 signaling pathway.

Corpus Striatum ; Dopaminergic Neurons ; Humans ; Mesencephalon ; Parkinson Disease/drug therapy* ; Pars Compacta ; Substantia Nigra

Objective:To investigate the predictive value of neutrophil-lymphocyte ratio (NLR) for Trousseau’s syndrome (TS) in patients with acute multiple cerebral infarctions (AMCI).Methods:The patients with AMCI in Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine from July 2013 to March 2022 were retrospectively enrolled. The demographic and baseline clinical data of patients with TS and those without TS were compared. Multivariate logistic regression analysis was used to determine the independent influencing factors of TS-AMCI, and receiver operating characteristic (ROC) curve was used to evaluate the predictive value of NLR for TS-AMCI. Results:A total of 59 patients with AMCI were enrolled, including 43 males and 16 females, aged 64.9±14.0 years. There were 16 patients in the TS-AMCI group and 43 in the non-TS-AMCI group. The proportions of patients with diabetes mellitus, hypertension and previous stroke or transient ischemic attack in the TS-AMCI group were significantly lower than those in the non-TS-AMCI group (all P<0.05), while the proportion of patients with ischemic heart disease were significantly higher than that in the non-TS-AMCI group ( P<0.05). The proportion of patients with bilateral infarction in the TS-AMCI group was significantly higher than that in the non-TS-AMCI group ( P<0.001). The D-dimer, NLR, white blood cell count, neutrophil count, monocyte count, percentage of neutrophils, total cholesterol and low-density lipoprotein cholesterol in the TS-AMCI group were significantly higher than those in the non-TS-AMCI group (all P<0.001), while the lymphocyte count, lymphocyte percentage, red blood cell count, hemoglobin and hematocrit were significantly lower than those in the non-TS-AMCI group (all P<0.001). Multivariate logistic regression analysis showed that high NLR was an independent predictor of TS-AMCI (odds ratio [ OR] 2.897, 95% confidence interval [ CI] 1.270-6.527; P=0.011), while high hemoglobin was independently negatively correlated with TS-AMCI ( OR 0.839, 95% CI 0.723-0.975; P=0.022). ROC curve analysis showed that the area under the curve of NLR for predicting TS-AMCI was 0.929 (95% CI 0.831-0.979; P<0.001). When the NLR cutoff value was 4.01, the corresponding Youden index was 0.744. At this time, the sensitivity and specificity were 100% and 74.42% respectively. Conclusion:NLR has high predictive value for TS-AMCI.

Objective:To detect the expression level of RAB39B gene and the effect of RAB39B on autophagy and α-synuclein, and then investigate the role of RAB39B gene mutation c.536dupA in the pathogenesis of Parkinson′s disease.Methods:Based on the novel RAB39B gene c.536 dupamutation identified in the previous work, the recombinant expression plasmid (pcDNA3.1-HA-RAB39B-536) of RAB39B gene with this mutation and wild-type recombinant expression plasmid (pcDNA3.1-HA-RAB39B) of RAB39B gene were constructed, and the recombinant expression plasmid was transfected into N2a cells with liposome as experimental group. The control group was made up with N2a cells transfected with plasmid pcDNA3.1-HA-RAB39B. Real-time polymerase chain reaction, Western blotting, immunofluorescence and immunoprecipitation techniques were used to detect the expression level of mutant RAB39B gene and the effects of RAB39B on autophagy and α-synuclein.Results:In the N2a cell model, the transcription level of mutant RAB39B was about twice that of wild type RAB39B, while the protein level of mutant RAB39B (0.30±0.00) was significantly lower than that of wild type (1.50±0.25, t=8.313, P<0.05). After adding proteasome inhibitor MG132, the protein level of mutant RAB39B increased (0.70±0.10, t=6.925, P<0.05); the level of microtubule-associated protein 1 light chain 3 BⅡ/Ⅰ of mutant RAB39B (3.11±0.30) was significantly lower than that of wild type (7.03±0.20, t=18.831, P<0.05); overexpression of wild type and mutant RAB39B did not affect the level of endogenous α-synuclein; overexpression of wild-type RAB39B resulted in elevated level of exogenous wild-type (p.A53T; from 0.60±0.11 to 1.25±0.08, t=8.254, P<0.05) and mutant (from 0.55±0.08 to 1.15±0.08, t=9.293, P<0.05) α-synuclein. Conclusions:The stability of the RAB39B protein decreased with the appearance of c.536 dupA mutation, the mutant protein may be degraded through the ubiquitin-proteasome pathway, and this mutation may affect the autophagy level of cells. RAB39B protein may interact with α-synuclein in vivo and may be involved in the maintenance of the stable level of α-synuclein.

Objective:To report the clinical, electrophysiological and genetic features in a Chinese family with distal hereditary motor neuropathy type V (dHMN-V) and screen the pathogenic mutant gene.Methods:A family with the history of inherited peripheral neuropathy was recruited in the First Affiliated Hospital of Zhengzhou University in July 2017. The clinical features and electrophysiological data were investigated. Genetic testing on well-established genes associated with hereditary peripheral neuropathy was conducted by targeted high throughput sequencing and the candidate variant was screened in the family and normal controls.Results:There were four affected individuals in the family. The proband, a 25-year-old male, was characterized by weakness and atrophy in the distal extremities primarily affected the upper extremities without sensory impairment. Electrophysiological study showed chronic neurogenic pattern in the upper and lower limb muscles. The motor conduction showed reduced velocity and compound muscle action potential amplitude, while the sensory conduction studies results were normal. The grandfather, a maternal uncle and a cousin of the proband exhibited similar clinical manifestations and electrophysiological abnormality. Genetic testing revealed a heterozygous mutation, c.880G>A(p.G294R), in the GARS gene in the proband. Proband′s mother and two other affected individuals carried the mutation which was confirmed by Sanger sequencing. The mutation site was not found in the unaffected members from the family and 300 unrelated normal controls. The variant is a novel mutation which has not been reported in dbSNP, ExAC and 1000 Genomes Project databases. Conclusion:The results suggest that the novel c.880G>A(p.G294R) mutation of the GARS gene is responsible for the Chinese patients with dHMN-V, and the findings broaden the mutational spectrum of GARS gene.

Objective The purpose of study was to evaluate the safety and effectiveness of theblood-letting and herbal-cupping therapy for lumbar spinal stenosis.Methods A multi-center prospective case series was performed.The LSS patients meeting the inclusion criteria received 8 treatments as a course and 4 courses in total.The primary outcomes were the symptom severity and physical function scale ofthe Swiss Spinal Stenosis Measurement (SSM,total score 0-5 for each domain).The secondary outcomes were thethe 12-item short form health survey (SF-12,total score 0-100),and Oswestry disability index (ODI,total score 0-100) at time of baseline,completion of last treatment of each course.The minimal clinically important differences (MCIDs) were calculated for estimating the percentage of improvement in the population.The adverse events were reported at any time of the intra-and post-operation.This was a phrase analysis of the studyat seven months.Results Forty-eight patientswere included,with 64.6％ (31/48) of LSS showing neurogenic claudication (walking distance ≤200 m).The average age was 63.1 ± 11.7 years,19 (39.6％) female,and the average BMI was 25.3 ± 3.3 kg/m2.The scores of symptom severity scale of SSM were 2.8 ± 0.6,2.6 ± 0.7,2.3 ± 0.6,1.9 ± 0.2 at baseline,1st,2nd,3rd course,and the scores of physical function scale were 2.5 ± 0.8,2.4 ± 0.7,2.1 ± 0.5,1.8 ± 0.3,and all the changes between baseline and each course showed significant improvement.The patient satisfaction of SSM,ODI and SF-12 showed significantimprovements after the 1st,2nd,3rd course (P＜0.05).The SF-12 subgroup physical composite scores after 3rd course and mental composite score after 1st showed no significant improvement.The minimal clinically important difference for the “SymptomSeverity scale” in the SSM was achieved withimprovement of 18.8％,40.6％,83.3％ in the LSS patient population after 1st,2nd,3rd course;and the "physical function scale" in SSM was achieved withimprovement of 22.9％,31.3％,50.0％.A total of 15 patients felt pain when they were micro-punctured with little blood at first time,but the symptom wereimmediately relieved without any treatment.Conelusions The Blood-letting and herbal-cupping therapy could benefit patients with lumbar spinal stenosis after third course of treatment in the fields of symptom relief and quality of life with no severe adverse event.However,this was a phrase analysis,so more evidence of this study and large comparative researches should be warranted in future.

Determining effective traditional Chinese medicine (TCM) treatments for specific disease conditions or particular patient groups is a difficult issue that necessitates investigation because of the complicated personalized manifestations in real-world patients and the individualized combination therapies prescribed in clinical settings. In this study, a multistage analysis method that integrates propensity case matching, complex network analysis, and herb set enrichment analysis was proposed to identify effective herb prescriptions for particular diseases (e.g., insomnia). First, propensity case matching was applied to match clinical cases. Then, core network extraction and herb set enrichment were combined to detect core effective herb prescriptions. Effectiveness-based mutual information was used to detect strong herb-symptom relationships. This method was applied on a TCM clinical data set with 955 patients collected from well-designed observational studies. Results revealed that groups of herb prescriptions with higher effectiveness rates (76.9% vs. 42.8% for matched samples; 94.2% vs. 84.9% for all samples) compared with the original prescriptions were found. Particular patient groups with symptom manifestations were also identified to help investigate the indications of the effective herb prescriptions.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Child ; China ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Propensity Score ; Sleep Initiation and Maintenance Disorders ; drug therapy ; Young Adult

Objective To investigate correlation between the rs12122341 polymorphism and ischemic stroke and its major subtypes in Chinese Han population.Methods The patients with ischemic stroke and matched healthy controls in Chinese Han population were enrolled.The rs12122341 genotype was detected by the improved multiple ligase detection reaction (iMLDR).Results A total of 776 patients with ischemic stroke (415 large artery atherosclerotic stroke and 361 small artery occlusive stroke) and 776 healthy controls were enrolled.Genotyping showed that only rs12122341 CC and CG genotypes were detected in all subjects,and no GG genotype was detected.There was no significant difference in frequencies of allele and genotype between the patient group and the control group.Multivariate logistic regression analysis showed that there were no significant correlations between rs12122341 polymorphism and ischemic stroke (odds ratio [OR] 1.482,95％ confidence interval [CI]0.641-3.421;P =0.447),large artery atherosclerotic stroke (OR 1.972,95％ CI 0.655-6.034;P=0.227),and small arterial occlusive stroke (OR 1.632,95％ CI 0.437-6.262;P =1.000).Conclusions There is no significant correlation between the rs12122341 polymorphism and risk of ischemic stroke and its major subtypes in Chinese Han population.

Objective To analyze the clinical manifestations and genetic mutations in 3 pedigrees with hereditary spastic paraplegia (HSP).Methods Three pedigrees diagnosed as having HSP in our hospital from January 2014 to November 2015,were chosen;the clinical manifestations,electrophysiology and imaging features of the patients in these three families were analyzed.Genomic DNA was extracted from peripheral venous blood,and the targeted gene capturing was employed to identify the disease-causing genes of these patients.Results The patients from the first family was familiar HSP,and the main clinical features were progressive lower limbs weakness and abnormal gait without cognitive impairment;the patients from the second family were familiar HSP and those from the third family were HSP without family history,and the main clinical features of the two pedigrees were slowly progressive spastic paraplegia and cognitive impairment.In addition,thin corpus callosum was visible in MR imaging of family three.Genetic testing showed the first family presented with a known mutation c.715C＞T ofA TL1 exon 7 and the loci co-segregated in the family.The second family presented with novel compound heterozygous mutations in the SPG11 gene:c.3099_3103delGTTTG mutation of exon 17and c3817 3818insTGA mutation of exon 22;novel compound heterozygous mutations in the SPG11 gene in the third family were detected as follows:c.6194C ＞G mutation of exon 32 and c.5121+1C＞T splicing mutation ofintro 29.Conclusions Four novel mutations in SPG11 gene and one known mutation in A TL1 gene are found,which enriches the known HSP mutation types.Targeted gene capture is an efficient and rapid tool for identifying the causation of some complex and genetically heterogeneous neurodegenerative diseases.

Objective To look for more serum biomarkers supporting the diagnosis of multiple system atrophy ( MSA) and providing more evidence for early treatment.Methods All patients and healthy controls were enrolled from January 2011 to March 2015 in the First Affiliated Hospital of Zhengzhou University.Demographic features and biochemical examination results were collected.The t test was used to compare the lipid levels between MSA patients and controls.LSD-t test was used to compare the lipid levels among subtypes of MSA patients.Multivariate Logistic regression analysis was conducted to analyze the influencing factors.The relevance between lipid levels and onset age, disease duration and Hoehn & Yahr stage was calculated by Spearman correlation coefficients.Results Participants included 195 MSA patients and 195 age-and gender-matched controls with no neurological diseases.The levels of total cholesterol ((4.33 ±0.90) mmol/L), triglyceride ((1.27 ±0.71) mmol/L), low-density lipoprotein (LDL;(2.70 ±0.76) mmol/L) were significantly lower in patients than in controls ((4.52 ±0.85), (1.47 ± 0.86), (2.85 ±0.71) mmol/L ,t=2.056,2.528 and 2.149 respectively, all P<0.05).The levels of total cholesterol ((4.28 ±0.96) mmol/L) and triglyceride ((1.20 ±0.64) mmol/L) were significantly lower in MSA-P patients than in control group ((4.52 ±0.85), (1.47 ±0.86) mmol/L;LSD-t=1.983, 2.566, both P<0.05).After adjusting for age, gender and histories, the odds ratio ( OR) was 0.31 (95%CI 0.15-0.65, P =0.002 ) for MSA patients in the highest quartile of triglyceride and 0.38 (95%CI 0.17 -0.83,P=0.016) for those in the highest quartile of high-density lipoprotein (HDL), compared with the lowest quartiles.And HDL level was in a significantly positive correlation with onset age (r=0.15, P=0.039).Conclusion Our data suggest that triglyceride and HDL may be associated with the prevalence of MSA, and the lower levels of HDL, the earlier onset of MSA.