Effects of NaHS on MBP and learning and memory in hippocampus of mice with spinocerebellar ataxia / 中国应用生理学杂志

To investigate the effects of exogenous NaHS on myelin basic protein (MBP) and learning and memory of hippocampal neurons in mice with spinocerebellar ataxia type 3 (SCA3) and its therapeutic significance. Twelve male normal mice were randomly selected as normal control group (NC Group), and 48 SCA3 mice were randomly selected as SCA3 model group (M Group), low dose group (NL Group, 10 μmol/kg), medium dose group (NM Group, 50μmol/kg) and high dose group (NH Group, 100 μmol/kg), 12 rats in each group. The drug treated groups were injected with NaHS intraperitoneally once a day for 4 weeks. The changes of learning and memory ability of SCA3 mice before and after the intervention of different doses of NaHS were determined by Morris water maze, the content of hydrogen sulfide (HS) in hippocampus was measured by spectrophotometry, the expression of MBP was detected by immunohistochemistry, and the morphological changes of neuron myelin sheath were observed by electron microscope. Compared with the control group, the learning and memory ability of SCA3 mice was decreased significantly (P＜0.05), and the content of HS in hippocampus was decreased (P＜0.05). After different doses of exogenous NaHS treatment, the learning and memory ability was improved in different degrees (P＜0.05), and the contents of HS and MBP in hippocampus of SCA3 mice were also improved in different degrees (P＜0.05). Exogenous NaHS may increase the contents of HS and MBP in the hippocampus of SCA3 mice, which may have a protective effect on the neurons, and then improve the learning and memory ability of SCA3 mice, and provide a new idea for the treatment of SCA3.

Morphologic evolution of recent small sub-cortical infarcts and adjacent white matter in the basal ganglia in a Chinese cohort / 中华医学杂志(英文版)

BACKGROUND@#Data on the evolution of recent small sub-cortical infarcts are limited, especially in the Chinese. Previous studies have reported a large heterogeneity in cavitation and infarct location; therefore, the present study assessed the morphology of small sub-cortical infarcts in the basal ganglia in a Chinese cohort.@*METHODS@#Patients who had experienced a recent, single, small sub-cortical infarct in the basal ganglia and received at least one follow-up magnetic resonance imaging (MRI) scan were retrospectively identified from January 2014 to June 2018. Time to follow-up imaging, baseline infarct size, vascular risk factors, and other clinical data, as well as the morphologic changes of the index infarct and surrounding white matter were recorded. Demographic, clinical and MRI characteristics were respectively compared among three groups (white matter hyper-intensitie [WMH] vs. cavitation vs. absent) and between with and without new WMH formation groups. In addition, logistic regression analyses were performed in investigating the determinate independent predictors for new WMH formation.@*RESULTS@#Seventy-eight subjects were included with a median follow-up time of 304 days (range: 124-552 days). We found a significant reduction in infarct size at follow-up: 46 of 78 (59.0%) infarctions showed some degree of cavitation, 19 of 78 (24.4%) index lesions resembled non-cavitated WMH, and 13 of 78 (16.7%) infarcts had disappeared at follow-up MRI. No factors were found to be associated with differential outcomes of the infarcts. In addition, 8 of 78 (10.3%) patients demonstrated new WMH formation surrounding the index infarct; white matter progression (odds ratio = 15.95, 95% confidence interval = 1.65-153.99; P = 0.017) was an independent risk factor of new WMH formation.@*CONCLUSIONS@#More than half of the small sub-cortical infarcts in the basal ganglia progressed to cavities, demonstrating that these infarcts can be reduced and go undetected. The presence of new WMH around the infarct may be indicative of the worsening progression of cerebral small vessel diseases. Additionally, white matter progression is an independent risk factor, which may be a potential therapeutic target.

Nine-hole Peg Test and Ten-meter Walk Test for Evaluating Functional Loss in Chinese Charcot-Marie-Tooth Disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The 9-hole peg test (9-HPT) and 10-meter walk test (10-MWT) are commonly used to test finger motor function and walking ability. The aim of this present study was to investigate the efficacy of these tests for evaluating functional loss in Chinese Charcot-Marie-Tooth (CMT) disease.</p><p><b>METHODS</b>Thirty-four Chinese CMT patients (CMT group) from August 2015 to December 2016 were evaluated with 9-HPT, 10-MWT, CMT disease examination score, overall neuropathy limitation scale (ONLS), functional disability score, and Berg Balance Scale (BBS). Thirty-five age- and gender-matched healthy controls (control group) were also included in the study. Student's nonpaired or paired t-test were performed to compare data between two independent or related groups, respectively. The Pearson test was used to examine the correlations between recorded parameters.</p><p><b>RESULTS</b>The mean 9-HPT completion time in the dominant hand of CMT patients was significantly slower than that in the healthy controls (29.60 ± 11.89 s vs. 19.58 ± 3.45 s; t = -4.728, P < 0.001). Women with CMT completed the 9-HPT significantly faster than men with CMT (dominant hand: 24.74 ± 7.93 s vs. 33.01 ± 13.14 s, t = 2.097, P = 0.044). The gait speed of the average self-selected velocity and the average fast-velocity assessed using 10-MWT for CMT patients were significantly slower than those in the control group (1.03 ± 0.18 m/s vs. 1.44 ± 0.17 m/s, t = 9.333, P < 0.001; 1.31 ± 0.30 m/s vs. 1.91 ± 0.25 m/s, t = 8.853, P < 0.001, respectively). There was no difference in gait speed between men and women. Both 9-HPT and 10-MWT were significantly correlated with the ONLS, functional disability score, and BBS (P < 0.05 for all).</p><p><b>CONCLUSION</b>The 9-HPT and 10-MWT might be useful for functional assessment in Chinese patients with CMT.</p>

Endothelial Progenitor Cells' Classification and Application in Neurological Diseases

The therapeutic effects of endothelial progenitor cells (EPCs) on ischemic stroke have been extensively studied in recent years. However, the differences in early EPCs and endothelial outgrowth cells (EOCs) are still unclear. Clarifications of their respective properties and specific functioning characteristics contribute to better applications of EPCs in ischemic diseases. In this review, we discuss cellular origin, isolation, culture, surface markers of early EPCs and EOCs and relevant applications in neurological diseases. We conclude that EOCs possess all haracteristics of true endothelial progenitors and have potent advantages in EPC-based therapies for ischemic diseases. A number of preclinical and clinical applications of EPCs in neurological diseases are under study. More studies are needed to determine the specific characteristics of EPCs and the relevant mechanisms of EPCs for neurological diseases.

Analyses of exercise-induced muscle damage-specific microRNA expression and molecular target of sarcolemmal damage in rats / 生理学报

In the present study, we were to screen the specific microRNA (miRNA) of exercise-induced muscle damage (EIMD) and assess the EIMD-specific miRNAs-regulated target of sarcolemmal damage in rats. Twenty-four male Sprague-Dawley (SD) rats were randomly divided into 3 groups, which included sedentary (C), 24 h post-exercise (E24) and 48 h post-exercise (E48) groups. Rat EIMD model was established by an acute eccentric exercise, i.e., a downhill running treatment at -16º gradient. EIMD characteristics were verified by Evans blue dye staining, differentially expressed miRNAs were detected by microarray assay, EIMD-specific miRNAs expressions were further validated by real-time quantitative RT-PCR (RT-qPCR), and targets of the miRNAs were predicted based on mRNA expressions of associated proteins and related pathway core molecules of sarcolemmal damage. Two EIMD-specific expressed miRNAs, including miR-206-3p and miR-139-3p, were found in the study. There was a significantly negative correlation (P < 0.05) between miR-206-3p expression and dystrophin (r = -0.68), utrophin (r = -0.64), JNK (r = -0.62) or ERK1 (r = -0.68) respectively, but no correlation was found between miR-139-3p and these biomolecules. The results suggest that: i) the expression profile of miRNAs in rat is significantly affected by EIMD, ii) miR-206-3p and miR-139-3p are the EIMD-specific miRNAs, and iii) miR-206-3p may control sarcolemmal damage by regulating dystrophin, utrophin, JNK and ERK1.

Nerve growth factor for the treatment of spinocerebellar ataxia type 3: an open-label study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Spinocerebellar ataxia type 3 (SCA3) is the most common subtype of SCA worldwide, and runs a slowly progressive and unremitting disease course. There is currently no curable treatment available. Growing evidence has suggested that nerve growth factor (NGF) may have therapeutic effects in neurodegenerative diseases, and possibly also in SCA3. The objective of this study was to test the efficacy of NGF in SCA3 patients.</p><p><b>METHODS</b>We performed an open-label prospective study in genetically confirmed adult (>18 years old) SCA3 patients. NGF was administered by intramuscular injection (18 μg once daily) for 28 days consecutively. All the patients were evaluated at baseline and 2 and 4 weeks after treatment using the Chinese version of the scale for assessment and rating of ataxia (SARA).</p><p><b>RESULTS</b>Twenty-one SCA3 patients (10 men and 11 women, mean age 39.14 ± 7.81 years, mean disease duration 4.14 ± 1.90 years, mean CAG repeats number 77.57 ± 2.27) were enrolled. After 28 days of NGF treatment, the mean total SARA score decreased significantly from a baseline of 8.48 ± 2.40 to 6.30 ± 1.87 (P < 0.001). Subsections SARA scores also showed significant improvements in stance (P = 0.003), speech (P = 0.023), finger chase (P = 0.015), fast alternating hand movements (P = 0.009), and heel-shin slide (P = 0.001).</p><p><b>CONCLUSIONS</b>Our preliminary data suggest that NGF may be effective in treating patients with SCA3.</p>

Reliability and validity of the Chinese version of the Scale for Assessment and Rating of Ataxia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The Scale for the Assessment and Rating of Ataxia (SARA) was shown to be a reliable and valid measurement for patients with spinocerebellar ataxia (SCA). The Brazilian version and the Japanese version of SARA were favorable for good reliability and validity. This study aimed to translate SARA into Chinese and test its reliability and validity in measurement of cerebellar ataxia.</p><p><b>METHODS</b>SARA was translated into Chinese. A total 39 patients with degeneration cerebellar ataxia were evaluated independently by two neurologists with the Chinese version of SARA. Then the patients were evaluated by one of above neurologists with International Cooperative Ataxia Rating Scale (ICARS). The statistical analyses were performed using SPSS 17.0 for Windows.</p><p><b>RESULTS</b>The Cronbach's alpha coefficient of the Chinese version of SARA was 0.78, which represents a good internal consistence. The correlation coefficient of the Chinese version of SARA scores between the two evaluators was 0.86, illustrating that the inter-rater reliability of Chinese version of SARA was good. The correlation coefficient between the Chinese version of SARA and ICARS was 0.91, illustrating that the criterion validity of Chinese version of SARA was not bad.</p><p><b>CONCLUSIONS</b>The Chinese version of SARA is reliable and effective for the assessment of degeneration cerebellar ataxia. Compared with ICARS, the evaluation of Chinese version of SARA is more objective, the assessment time is shortened, and the maneuverability is better.</p>

Change in gastrocnemius dystrophin and metabolic enzymes and increase in high-speed exhaustive time induced by hypoxic training in rats / 生理学报

The aim of the present study was to explore the changes and roles of dystrophin and membrane permeability in hypoxic training. Seventy-two 8-week-old Sprague Dawley (SD) rats were randomly divided into 4 groups, normoxic non-train (NC), normoxic train (NT), hypoxic non-train (HC), and hypoxic train (HT) groups. The rats of each group were randomly divided into three subgroups, non-exhaustive, low-speed exhaustive test and high-speed exhaustive test subgroups. Rats in hypoxia groups lived and were trained in a condition of 12.7% oxygen concentration (equal to the 4 300 m altitude). NT and HT groups received 4 weeks of training exercise. Then the rats in all non-exhaustive subgroups were sacrificed, and gastrocnemii were sampled for the measurements of lactate dehydrogenase (LDH), succinatedehydrogenase (SDH), malate dehydrogenase (MDH) activities. Moreover, serum LDH activity was analyzed. Low-speed exhaustive test and high-speed exhaustive test subgroups received exhaustive tests with 20 (71% VO2max) and 30 m/min speed (86% VO2max), respectively, and their exhaustive times were recorded. The results showed that, compared with normoxic groups, the weights in hypoxia groups exhibited slower increase. The level of dystrophin in HT group without exhaustion test didn't change significantly. The muscle MDH activities were markedly affected by the different oxygen concentration, training and their interaction (P < 0.05), whereas the muscle LDH activities were only affected by the different oxygen concentration (P < 0.05). Serum LDH activities were affected by the interaction of the different oxygen concentration and training (P < 0.05), showing decreased muscle LDH and increased blood LDH activities. The exhaustion time were markedly affected by the different test speed, training and their interaction (P < 0.05), and also affected by the interaction of the different oxygen concentration and training (P < 0.05), but didn't affected by oxygen concentration. The exhaustive time of HT high-speed exhaustive test subgroup was more than NT high-speed exhaustive test subgroup in 30 m/min exhaustion test. Compared with NT high-speed exhaustive test subgroup, HT high-speed exhaustive test subgroup had an earlier fatigue in the test, but had a rapid recovery. These results suggested that hypoxic training can effectively increase the rats' high-speed exhaustive time. The mechanism may be related to an increase in serum LDH caused by the increased membrane permeability after hypoxic training.

Sex-dependent association of phosphodiesterase 4D gene polymorphisms with ischemic stroke in Henan Han population / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recent evidence has implicated the gene for phosphodiesterase 4D (PDE4D) as susceptibility gene for ischemic stroke (IS) in Icelandic population. However, there are few reports on the associations between PDE4D gene polymorphisms and IS in Chinese individuals. The present study aimed to investigate the possible association of genetic polymorphisms in PDE4D gene with IS in Henan Han population.</p><p><b>METHODS</b>A total of 400 patients with IS and 400 matched controls were examined using a case-control design. Two single nucleotide polymorphism (SNPs) (rs918592 and rs2910829) in PDE4D gene were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Odds ratios (OR) and 95% confidence intervals (95%CI) were calculated to test the association between the genetic factors and IS. Genetic parameter and association studies were carried out with SPSS 16.0.</p><p><b>RESULTS</b>Among the two SNPs tested, the rs918592 was significantly associated with IS (OR: 1.351, 95%CI: 1.110 - 1.645), especially in male patients (OR: 1.427, 95%CI: 1.105 - 1.844). Haplotype analysis showed that A-T was associated with an increased risk of the IS (OR: 2.114, 95%CI: 2.005 - 2.230) while G-T was associated with decreased risk of IS (OR: 0.419, 95%CI: 0.302 - 0.583). Protecting effect of haplotype G-T was also significant in males (OR: 0.264, 95%CI: 0.162 - 0.431).</p><p><b>CONCLUSIONS</b>The present study demonstrated a strong association of rs918592 with IS. Haplotype A-T increased the risk of IS while haplotype G-T had a protective effect in Henan Han population. The association was sex-dependent with male patients showing stronger effect.</p>

Construction of a recombinant BAC-HSV-1 strain HF with a GFP reporter gene and characterization of its infectious progeny virus / 病毒学报

To construct the plasmid of BAC-HSV-1 with GFP reporter gene and research the biological property of its infectious progeny virus. We constructed the plasmid C223-UL43-left-arms-UL47-right-arms which carried the homologous sequences of HSV-1. Liposome embedding method was used to transfect HSV-1 genome and the plasmid C223-UL43-left-arms-UL47-right-arms linearized by Mlu I digestion into Vero cells. After the successful homologous recombination in the eukaryotic cells, the recombinant BAC-HSV-1 with GFP reporter gene was generated. Then, the positive CPE were taken by plaque purification and by hirt extraction during the moment of the circularization of HSV-1 DNA, and the plasmid of BAC -HSV-1 was acquired. Electroporation was used to transfect the BAC -HSV-1 into DH10B, and then the single colonies of interest were confirmed both by MluI digestion and PCR. Experimental group and the control group cells were given BAC-HSV-1 plasmid and HSV-1 genomic DNA respectively to produce the BAC-HSV-1 and HSV-1 progeny virions. Vero cells were inoculated with the progeny virions at MOI = 0.1 and then a TCID50 assay was performed to determine the titers of virons in the two groups at 48 hours post inoculation. The plasmid BAC-HSV-1 was successfully constructed by the restriction enzyme analysis and the PCR. The titers of progeny virions were calculated by the TCID50 assay. No significant difference in the titers of virions between two groups was observed (P > 0.05). The infectious BAC-HSV-1 shuttle virus/plasmid between eukaryotic and prokaryotic cells was successfully constructed.

Construction of the HSV-1 strain HF amplicon and study on its unversal function between different HSV serotypes / 病毒学报

The study aimed to construct the amplicon vector of HSV-1 strain HF and explore its universal package function between different serotypes of HSV. OriS and pac elements were obtained by enzyme digestion from the Plasmid BAC-HSV-1 strain HF and sequenced. With red fluorescence (DsRed) as a reporter gene, the amplicon vector of HSV-1 strain HF was constructed based on pSilencer2.0-U6. The amplicon vector was transfected into Vero cells by lipofectamine 2000, then packaged by HSV-1 strain HF and HSV-2 strain HG52 as helper virus separately. The supernatant was collected after cytopathic effect. Red fluorescence was observed in Vero cells reinfected by the supernatant. In this study,the amplicon vector of HSV-1 strain HF was successfully constructed and it could be packaged by HSV-1 strain HF and HSV-2 strainHG52.

SiRNA targeting ICP4 attenuates HSV-1 replication / 病毒学报

HSV-1, a neurotropic virus, always leads to severe nervous symptoms. It is hard to completely eradicate the latent viruses after conventional therapy so that recurrence is inevitable. ICP is a key regulator for HSV replication and transcription that determines the cytolytic infection or latent state. In search of new anti-virus strategy targeting HSV-1ICP4, two pairs of siRNA were designed, and a recombinant eukaryotic lentiviral expression plasmid pLKO-puro(r)-hU6-siRNA was constructed. Vero cells were transfected with the designed siRNAs by Lipofectamine 2000 and four stable monoclonal cell lines were established by puromycin screening method. The ICP4 expression at mRNA level was detected with real-time PCR, and the HSV-1 replication was measured with TCID50 assay. SiRNA was shown as an effective way to inhibit the expression of ICP4 in monoclonal cell lines. Meanwhile, HSV-1 replication was significantly inhibited when ICP4 was shut down by siRNA. We conclude that siRNA targeting ICP4 attenuates HSV-1 replication. Further more, multi-site siRNAs show stronger inhibitory effect on viral replication, which may be an effective and feasible approach for biological anti-viral drugs.

Changes of dystrophin and desmin in rat gastrocnemius under micro-damage induced by hypoxia / 生理学报

To explore the changes and regulation mechanism of dystropin and desmin under muscle injury without mechanic stress, 40 male Sprague-Dawley rats were randomly divided into 5 groups, which included normoxia control and hypoxia groups for 1, 2, 4 and 7 d with 10% O2. Two rats from each group were examined for sarcolemma integrity using Evans blue dye (EBD) and EBD-positive fiber typing by metachromatic dye-ATPase method. The rest six rats from each group were analyzed for the changes of protein content and gene expression using Western blot, RT-PCR and fluorescence assays. The results showed that the EBD-positive muscle fibers, mainly type IIA and type IIB, appeared at 1 d after hypoxia exposure. Both the ratio of EBD-positive cell and the mean fluorescence density were significantly higher in hypoxia groups than those in control group (P<0.05). The contents of dystrophin and desmin fluctuated after hypoxia exposure, increased at 1 d, decreased at 2 d, increased dramatically again at 4 d, and returned to a normal level at 7 d. Consistently, the gene expression began to increase significantly after 2 d. The total activity of calpain was significantly higher in hypoxia groups at 1, 4 and 7 d. Significantly higher levels of HSP70 and HSP90 were also observed at 4 and 7 d, respectively (P<0.05). These results suggest that the mechanical stress is not the only cause of damage of sarcolemma membrane integrity. In contrast to eccentric contraction, hypoxia-induced muscle damage is not accompanied by the loss of dystrophin and desmin. The types of muscle fibers recruited by motor units and the activities of calpain may be important in hypoxia-induced damage of sarcolemma membrane integrity.

Effects of estrogen on P-Tau, ChAT and nerve growth factor protein expressions in the brain tissue of rats with Alzheimer's disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To examine the effect of estrogen on the expressions of phosphorylated Tau (P-Tau), ChAT and nerve growth factor (NGF) protein in the brain tissue of rat models of Alzheimer disease (AD).</p><p><b>METHODS</b>Rat models of AD were established by injecting Aβ1-42 protein fragments in the right lateral ventricle. Two weeks later, 17β-estradiol tablets were implanted subcutaneously at the neck of the rats and maintained for 30 days. The pathological changes in the rats' brain neurons and alterations in the expressions of P-Tau, ChAT and NGF proteins were observed using HE staining and immunohistochemistry, respectively.</p><p><b>RESULTS</b>In the AD rats, neurofibrillary tangles occurred in the brain tissue, and estrogen treatment significantly reduced the formation of neurofibrillary tangles. Estrogen treatment also resulted in lowered P-Tau expression and increased ChAT and NGF protein expressions in comparison with those in the AD model rats.</p><p><b>CONCLUSION</b>Estrogen can up-regulate ChAT and NGF and down-regulate tau protein expression, thus producing obvious therapeutic effect on AD in rats.</p>

Analysis for clinical and genetic characteristics of a sporadic FFI case / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To report and study a case of sporadic family fatal insomnia (SFFI) on its.</p><p><b>METHODS</b>Investigate clinical characteristics and family disease history of a suspect FFI patient. His clinical characteristic was analyzed, he and his 14 family members genomic DNA was extracted by standard techniques from their and blood detected with polymerase chain reaction (PCR) method and DNA sequencing to find out his prion protein (PrP) gene mutation. The patient's CSF was detected with Western-Blot method for 14-3-3 brain protein.</p><p><b>RESULTS</b>The patient was diagnosed as an sporadic FFI by his developed sleep disturbance and changes in sleep-awake rhythm, motor abnormalities, mental disorder, dementia, autonomic dysfunction; his family history; his 14-3-3 brain protein-positive (CSF) and analysis results of his PrP gene (codon point mutation D178N and methionine homozygosity at position 129M/M). Suggesting that in the future to identify CJD and FFI patients, screening should focus on clinical symptoms and laboratory results. The PrP gene of 14 family members did not appear Mutation, and there is no person suffering from the same disease.</p><p><b>CONCLUSIONS</b>The case was diagnosed as a sporadic familial fatal insomnia. Analysis of suspicious patients' genomic DNA for PrP gene mutation might be very important for FFI diagnosis because there exist many difficulties in clinical laboratory evaluation. This patient might be the first SFFI patient reported in China and the case finding might have momentousness in clinical and basical study.</p>