1.Application of miRNA-34a, miRNA-34c and miRNA-135a in early diagnosis of mild cognitive impairment and Alzheimer's disease
Lifang CHEN ; Lu ZHANG ; Manfu HAN ; Junming YIN ; Shiyu HU ; Zhuoyuan LIANG
Journal of Chinese Physician 2017;19(5):683-686
Objective To investigate the clinical value of miRNA-34a,miRNA-34c,and miRNA-135a in the early diagnosis of mild cognitive impairment (MCI) and Alzheimer's disease (AD).Methods The patients were collected in the outpatient and inpatient of Neurology during Aug 2014 to May 2016.The levels of miRNA-34a,miRNA-34c,an dmiRNA-135a were detected with quantitative real-time polymerase chain reaction (qRT-PCR).Results AD patients had higher level of miRNA-34a than the controls.The levels of miRNA-34c were higher in MCI and AD patients,while lower levels of miRNA-135a compared to the controls.Conclusions The miRNA 34a and miRNA-135a were likely to became the biomarker in early diagnosis of MCI and AD.
2.Prevalence and risk factors of stroke:findings from a community in Shenzhen, China
Yanxia ZHOU ; Manfu HAN ; Lijie REN ; Feng CHI ; Xiang TANG ; Xia LONG
International Journal of Cerebrovascular Diseases 2016;24(8):716-721
Objective To explore the prevalence and risk factors of stroke in a population over 40 years in a community in Shenzhen, China. Methods The subjects investigated were a population ≥40 years old in this cross-sectional study. Cluster sampling method was used to conduct the unified questionnaires, physical examination, and laboratory examination in the community residents, and then the survey data were used for online entry analysis. Results A total of 5 308 community residents were screened, and 160 experienced stroke. The crude prevalence of stroke was 3 014. 32/100 000. The prevalence of stroke increased with age, and that in males was significantly higher than that in females (3 721. 37/100 000 vs. 2 552. 93/100 000; χ2 = 5. 923, P = 0. 015). There were significant differences in the proportions of males, obvious overweight, hypertension, diabetes mellitus, atrial fibrillation, lack of physical activity, and family history of stroke, as well as age, systolic blood pressure, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and fasting glucose level between the stroke population and the non-stroke population (all P < 0. 05). Multivariate logistic regression analysis showed that hypertension (odds ratio [OR] 1. 737, 95% confidence interval [CI] 1. 161-2. 600; P = 0. 007), diabetes (OR 1. 917, 95% CI 1. 209-3. 040; P = 0. 006), atrial fibrillation (OR 1. 699, 95% CI 1. 113-2. 592; P = 0. 014), family history of stroke (OR 1. 585, 95% CI 1. 126-2. 231; P = 0. 008), advanced age (OR 4. 645, 95% CI 1. 868-11. 551; P = 0. 001), and physical inactivity (OR 4. 921, 95% CI 3. 552-6. 187; P < 0. 001) were the independent risk factors for stroke, and lack of physical activity was an independent protective factor for stroke. The proportion of smoking in males was more than that in females in all ages(all P < 0. 05). The proportions of hypertension (P < 0. 001) and hyperlipidemia (P < 0. 001) were gradually increased with age, and physical exercise was gradually reduced with age (P = 0. 001, except for subjects > 80 years) in both males and females. The proportions of diabetes (P < 0. 001) and atrial fibrillation (P < 0. 001) in males, and obvious overweight (P = 0. 001) in females were gradually increased with age, and the proportion of smoking in males weas gradually reduced with age. The proportions of hypertension (P < 0. 001) and obvious overweight (P < 0. 001) in males were significantly higher than those in females at the age of 40 to 49 years. The proportions of hypertension (P < 0. 001), diabetes (P < 0. 001) and obvious overweight (P < 0. 001) in males were significantly higher than those in females at the age of 50 to 59 years. The proportion of hypertension in males was significantly higher than that in females at the age of 60 to 69 years (P = 0. 039). The proportions of hypertension (P = 0. 016), atrial fibrillation (P = 0. 028) and hyperlipidemia (P = 0. 023) in females were significantly higher than those in males at the age of 70 to 79 years. The proportion of obvious overweight in females was significantly higher than that in males at the age of ≥80 years (P =0. 001). Conclusions The crude prevalence of stroke is higher among the community residents. The the levels of exposure to stroke risk factors including hypertension, diabetes and atrial fibrillation are higher. It may be important to intervene on these risk factors in community residents, especially in elders and those with family history of stroke.
3.Functional anatomy of the brain in depression patients with cognitive dysfunction based on functional magnetic resonance imaging
Jun LI ; Lijie REN ; Manfu HAN ; Zhuqing ZHOU ; Yuezhi LI
Chinese Journal of Neuromedicine 2014;13(2):142-145
Objective To investigate the characteristics of functional anatomy of the brain in depression patients with cognitive dysfunction and its pathological basis.Methods Twenty-four patients with first episode of depression,admitted to our hospital from May 2008 to December 2009,and 24 healthy controls,collected at the same time period,were chosen in our study; they were all assessed by Hamilton depression (HAMD) scale and Wisconsin test; fractional anisotropy (FA) values of the various parts of white matter were measured by diffusion tensor imaging (DTI),and functional MRI was performed when the testers were conducting N-back task; the related indicators and imaging results were statistically analyzed.Results The brain areas in the depression group having significant difference as compared with those in the healthy controls (P<0.05) included the bilateral frontal lobes (the leg bilateral frontal lobe,t=-5.884; the right bilateral frontal lobe,t=-4.517),anterior cingulate cortex (t=-14.402) and corpus callosum (t=-3.701).In patients of depression group,the left frontal FA values were negatively correlated with the disease duration (r=-0.555),and FA values of left bilateral frontal lobe and anterior cingulate cortex were negatively correlated to perseverative responses errors (Rpe) and mean response times (mRT,P<0.05); the disease duration was positively correlated to Rpe and mRT (P<0.05).The activating reduced brain areas of the depression group when they were conducting N-back task included bilateral frontal gyrus,left middle frontal gyrus,inferior fiontal gyrus and superior parietal lobule.Conclusion The depression patients have significant cognitive impairment; the damage of frontal white matter fiber and abnormalities of gray matter may be the pathological basis.
4.The characteristics of cognitive impairment in depression: a combined functional magnetic resonance and event-related potential study
Jun LI ; Lijie REN ; Manfu HAN ; Yuezhi LI ; Zhuqing ZHOU
Chinese Journal of Behavioral Medicine and Brain Science 2013;22(11):985-988
Objective To investigate the characteristics and relative pathogenesis of cognitive impairment in people with depression.Methods 24 people with depression and 24 healthy controls were evaluated respectively with HAMD scale,the WCST test,N-BACK task P300 and fMRI.Results (1) The WCST scores,N-back reaction (MRT),the P300 incubation period in depression group were significantly different from those in control group(P300 wave amplitude(4.12± 1.51) μV vs (6.42± 1.73) μV ; P300 latency(392.02±23.60) ms vs (309.43± 21.39) ms,t=4.922,P<0.01 ; t=12.726,P<0.01).(2) The illness course had positive correlation with Rep(r=0.596,P<0.01) and mRT(r=0.518,P<0.01).The P300 latency had positive correcation with Rpe(r=0.929,P< 0.01) and mRT(r=0.939,P<0.01).(3)Compared with control group,the decreased activation area in patients with depression were as follows:bilateral frontal gyrus,left middle frontal gyrus,inferior frontal gyrus and superior parietal lobule.Conclusion The depressive patients exist cognitive impairment mainly in frontal lobe.The longer with the illness,the wose with the impairment.P300 incubation period is a sensitive indicator of the frontal executive function.
5.Depression tensor imaging study of depression patients mood processing disorders
Lijie REN ; Jun LI ; Yuezhi LI ; Manfu HAN ; Yufeng SHAO ; Zhuqing ZHOU
Chinese Journal of Behavioral Medicine and Brain Science 2013;22(9):807-809
Objective To investigate the characteristics of the depression during emotional processing.Methods 24 participants with first episode of depression and healthy controls were assessed with HAMD scale,using DTI to dectect values of white matter FA,and using fMRI with pictures of emotional stimuli;thus results related with imagings were produced.The results were statistically analyzed.Results The brain areas indicating FA values deference with statistial significance in depression patients compared with the control ones included:left and right frontal lobe (left frontal lobe depression group 0.324 ± 0.090,control group 0.467 ± 0.072,P < 0.01),corpus callosum knee (depression group 0.614 ±0.146,control group 0.734 ±0.063,P<0.01),anterior cingulate gyrus (depression group 0.222 ±0.035,control group 0.343 ±0.021,P<0.01) ;the fronal FA values in depression grop were negatively correlated with the duration of bilateral frontal white matter (r =-0.555,P < 0.01).The activation of emotional brain regions stimulated by pictures includes frontal cortex-subcortical reticular system,and the hypothalamus and limbic system.There was a significant difference between two groups.Conclusion There may be abnormal emotional processing dystunction in patients with depression.It may be the pathological basis to have frontal white matter fiber tracts broken.
6.Phenotype in 6 patients with mitochondrial DNA G13513A mutation
Zhaoxia WANG ; Danhua ZHAO ; Xiaokun QI ; Manfu HAN ; Liqun FENG ; Yun YUAN
Chinese Journal of Neurology 2011;44(5):322-326
Objective To report 6 Chinese patients with mitochondrial encephalomyopathy caused by mitochondrial DNA(mtDNA)G13513A mutation and discuss the mitochondrial phenotype associated with this mutation based on the data of our patient series as well as the reports by others.Methods Direct sequencing of polymerase chain reaction(PCR)products or PCR-RFLP analysis Was performed to screen mtDNA G13513A mutation in 35 cases with mitoehondrial encephalomyopathy.who carried no mtDNA common mutations(1arge 8eale deletion,A3243G,T3271 C,A8344G,or T8993G/C).The clinical features,MRI changes were retrospectively collected and analyzed.Published studies of all patients with mtDNA G13513A mutation were also reviewed.Results Six patients were identified carrying mtDNA G13513A mutation.All patients presented stroke-like episodes with hemianopsia.hemiparesis or hemiparesthesia.Three adult patients presented clinical and radiological features of adult-onset mitochondrial myopathy,encephalopathy,lactic acidosis,and stroke-like episodes(MELAS),including stroke-like episodes,epilepsy,headache,short stature,sensorineural deafness,multifocal lesions on parietal,occipital and temporal lobes on cranial MRI scans.Three iuvenile.onset patients presented the clinical and brain MRI features of MELAS-Leigh syndrome(LS)overlap syndrome.In addition to the stroke-like episodes,they also showed brain stem lesions with dysarthria,ataxia,and ophthalmopJegia. Brain MRI revealed asymmetrical lesions in the cortex of the oecipital and temporal lobes,as well as symmetrical lesions in the bilateral basal ganglia and brainstem.Muslce biopsy showed ragged redfibem in 5 patients.The infant-onset LS or Leigh-like syndrome with mtDNA G135 13A was described in the English literature.Conclusions mtDNA G13513A mutation is a common pathogenic mutmion for mitochondrial encephalomyopathy,which can result in Leigh syndrome,MELAS-LS overlap syndrome and adult MELAS.The onset of various phenotypes is relatively age-dependent.
7.Correlation between fractional anisotropy values of diffusion tensor imaging and working memory impairment in patients with depression
Lijie REN ; Bingxun LU ; Mingxiang WU ; Manfu HAN ; Jing LI
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(4):325-327
Objective To investigate whether there are correlations between working memory impairmentand fractional anisotropy values and to explore the neuropathology underlying that the patients of depression suffered from memory impairment.Methods Thirty depression patients and 30 healthy controls group-matched by age,educational level were conducted the study.Mean correct reaction time(mRT)was recorded when they performed a One-Back Working Memory Task and fractional anisotropy(FA)values was recorded when they performed the diffusion tensor imaging.Statistics analysis was done respectively for mRT and FA values between two groups.Results Relative to mean correct reaction time((612.45±54.08)ms)of controls,the mean correct reaction time ((720.25±57.02)ms)of patients with depression was much longer(P<0.05)and the patients with depression had a lower FA values in the white matter of both frontal lobe,anterior cingulate gyrus,supramarginal gyrus,splenium of corpus callosum(P<0.05),and the FA values in the white matter of both frontal lobe were significantly negative correlated with mRT(r=-0.604,P<0.05).Conclusion The impairment of white matter region may be one of the neuropathology underlying that the depression patients suffer from memory impairment.
8.Mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes/Leigh overlap syndrome caused by mutation of mitochondrial DNA G13513A
Manfu HAN ; Runtao BAI ; Hongye FENG ; Weiyi TAO ; Zhaoxia WANG ; Yun YUAN
Chinese Journal of Neurology 2009;42(4):248-252
Objective To describe the chnical, neuroimagine, pathological and genetic features in a case with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS)/Leigh overlap syndrome.Methods The ease was a 22-year-old woman with recurrent headache, loss of visual acuity and general seizures over 11 years.MRI demonstrated symmetrical high T2-weighted signals in occipital and parietal lobes, in the late stage of the disease, the above imagine changes on MRJ were also shown in the bilateral basal ganglion and brainstem.She died of status epilepticus at age of 22.Brain autopsy and mitochondrial DNA (mtDNA) analysis were performed in the patient.Results The main neuropathological findings were muhifocal and lamilar spongiform in the cortex of the whole brain, the basal ganglion and middle brain.Gliosis, macrophagie reaction and capillary endothelial proliferation were observed in these areas.All 6 layers of the cortex and subcortical white matter in occipital and parietal lobes were severely affected.GI3513A mutation was found in the gene of mitochondria encoded NADH dehydrogenase subunit 5 (MTNDS).Conclusions MELAS/Leigh overlap syndrome presents the symptoms predominantly affecting the cerebral cortex.Neuroimagines suggested that the lesion initially involves the cerebral cortex and in the late stage implicates the basal ganglion and the brainstem, possibly caused by pathological changes of spongiform with capillary proliferation in these areas.
9.Glial cell-derived neurotrophic factors combined with transforming growth factor-beta 1 for in vitro differentiation of neural stem cells from rat spinal cord
Mingyong GAO ; Jiande XIAO ; Zhenyu LI ; Hongyin YAN ; Runtao BAI ; Manfu HAN
Chinese Journal of Tissue Engineering Research 2007;11(24):4856-4860
BACKGROUND: Glial cell-derived neurotrophic factor (GDNF) and transforming growth factor-beta 1 (TGF-β1)co-subordinate to TGF-β family. Both of them play very important roles in the development and differentiation of central and peripheral nervous system, and regulation of cell cycle in mammals.OBJECTIVE: To observe the differentiation of spinal cord-derived neural stem cells(NSCs) induced by GDNF combined with TGF-β1, and make a comparison of differentiation results with GDNF or TGF-β1 culture fluid.DESIGN: Controlled observation.SETTING: Central Laboratory, Shenzhen Hospital Affiliated to Southern Medical University.MATERIALS: Ten SD rats of clean grade, which were at conception for 16 days, were provided by the Experimental Animal Center, Tongji Medical College, Huazhong University of Science and Technplogy. Main reagents and materials:DMEM/F12,B27(GIBCO); basic fibroblast growth factor (bFGF), GDNF; TGF-β1(PeproTech);fetal bovine serum (FBS,Hyclone); nestin multiple antibody (Boster, Wuhan); glial fibrillary acidic protein (GFAP) multiple antibody; neurofilament protein (NF-200) monoclonal antibody (Sigma).METHODS: This experiment was carried out in the Central Laboratory, Shenzhen Hospital Affiliated to Southern Medcial University between October 2005 and September 2006. Under the aseptic condition, rat fetus was isolated for isolation and culture of spinal cord-derived neural stem cells. In this study, five groups were divided: basal medium group, control group, bFGF group, TGF-β1 group, GDNF+ TGF-β1 group. In the basal medium group, DMEM/F12 containing penicillin,streptomycin, amphotericin (AMPH) B and 0.02 volume fraction of B27 annex solution. At 1 week after primary culture, rat spinal cord-derived NSC clones proliferated in vitro stably were harvested. In the control group, 0.1 volume fraction of FBS was added into basal medium. In the later three groups, induced medium was exchanged, i.e. 20 μg/L bFGF, 2 μg/L TGF-β1, and 10 μg/L GDNF+2 μg/L TGF-β1 were added into the basal medium, respectively. ①The differentiation of spinal cord-derived NSCs induced by different factors were observed under the optical microscope. ②The expressions of neurons and astrocytes were detected by immunocytochemical staining labeling. ③ The differentiated cells were counted by sorting technique by means of fluorescence excitation flow cytometer, and the percentage of NSCs differentiating into neurons and astrocytes were detected under the different induction environments.MAIN OUTCOME MEASURES: ① Morphological feature of cell differentiation in each group. ② Immunohistochemical detection of NSCs in each group. ③ The percentage of NSCs differentiating into neurons and astrocytes in each group.RESULTS: ① Cell morphology during differentiation: At the early stage of differentiation, lots of cells creeped to all the directions, and one week later, most of the migrated cells adhered to the wall entirely. Neuron-like cells, astrocyte-like cells and oligodendrocyte-like cells could be identified in the low-density cell region. ②Immunohistochemical detection results: A lot of GFAP- positive astrocytes were found in the control group and TGF-β1 group; Many differentiated neurons and NF-200 staining positive were found in the bFGF group and GDNF+ TGF-β1 group. ③Percentage of stained neuron and astrocyte: at one week of induction, the percentage of stained neurons was higher in the GDNF+ TGF-β1 group than in the control group, bFGF group and TGF-β1 group (x2=24.15,19.56,25.32,P < 0.05-0.01), and the percentage of stained astrocytes was lower in the GDNF+ TGF-β1 group than in the control group, bFGF group and TGF-β1 group (x2=24.45,23.79,P < 0.01 ).CONCLUSION: The combined in vitro induction of GDNF and TGF-β1 contributes to the neuronal differentiation of spinal cord-derived NSCs, indicating that both of them have synergistic effect.
10.Effect of acrylamide on the degeneration and regeneration of rat myelinated fiber after sciatic nerve crush injury
Qiuyue HE ; Manfu HAN ; Mingli RAO
Chinese Journal of Tissue Engineering Research 2005;9(42):183-185
BACKGROUND:Ola rats is a kind of rats with genovariation, who displays Wallerian degeneration after peripheral neuroaxonal damage that is slower than that normal 6J rats, thereby additional damage factor may help fully understand the property of Ola rats.OBJECTIVE: To observe the effect of acrylamide on the degeneration and regeneration of sciatic nerve medullated fibers following crush injury of C57BL/Ola (Ola) rat and C57BL/6J (6J) rat.DESIGN: Randomized controlled experiment.SETTING: Department of Neurology,Second People's hospital of Shenzhen; Department of Neurology of First hospital of Jilin University MATERIALS: This experiment was carried out in the neurological department in the University of Occupational and Environmental, Japan from January to June 1996. Twelve adult Ola rats and 6J rats were adopted and evenly randomly divided into experimental group and comparison group.METHODS: Rats were subjected to general anaesthesia, and then the proximal section of sciatic nerve was exposed and frustrated with hemostatic forceps for 10 s before suture. Rats in the experimental group were given intraperitoneal injection of acrylamide in a total dosage of 350 mg, which replaced by the same volume of physiological saline in comparison group.At 14 days after sciatic nerve torsion injury, all rats were anaesthetized again and the distal section of sciatic nerve was obtained and cut into slices, meanwhile the cross sectional area, the density and size frequency distribution of medullated fibers, as well as the number of medullated fibers in each nerve were determined.MAIN OUTCOME MEASURES: The density and size frequency distribution of sciatic nerve medullated fibers, as well as the number, the maximum diameter and the mean diameter of medullated fibers in two group of 0la rats and 6J rats.RESULTS: Totally 12 Ola rats and 6J rats entered the result analysis.① No Ola rat displayed Wallerian degeneration; But medullated fiber degeneration and following neonatal small diameter medullated fibers could be observed in 6J rats. ②In the experimental group, the total density of sciatic nerve medullated fibers in 6J rats was lower than that of Ola rat (P < 0.05) ;with the total number of medullated fibers in 6J rats also less than that of Ola rat (P < 0.01 ), which predominated by obviously reduced big diameter fibers (P < 0.01); The mean diameter of medullated fiber in 6J rats was also obviously smaller than that of 0la rat (P < 0.01 ).CONCLUSION: The Wallerian degeneration is extremely slow in Ola rat after torsion injury, which cannot be affected by acrylamide; while acrylamide has obvious inhibition on the axonal neogenesis in 6J rat after torsion injury.

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