The clinical and genetic studies in a family of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical features and genetic diagnostic method of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL).</p><p><b>METHODS</b>A systematic study on the clinical manifestations, neuroimaging characteristics, therapeutic measures and molecular genetics was performed. An investigation on the onset and hereditary pattern of the family was also done.</p><p><b>RESULTS</b>The main clinical features including poor memory and history of stroke were found. And no risk factors of hypertension and arteriosclerosis were found. A positive family history was confirmed. Neuroimaging examination showed multiinfarct lesions and leukoencephalopathy. All these features are in conformity with those of CADASIL. A mutation in the third and fourth exon of the NOTCH3 gene was identified in the 10 cases of 4 generations. The clinical or subclinical onset in the 10 cases was consistent with classical autosomal dominant inheritance.</p><p><b>CONCLUSION</b>The clinical and molecular genetic features of the family accord with CADASIL.</p>

Establishment of spinal muscular atrophy cell model by RNAi / 中南大学学报(医学版)

OBJECTIVE@#To establish spinal muscular atrophy (SMA) cell model by blocking the expression of SMN1 gene with shRNA.@*METHODS@#The recombinant SMN1 shRNA expression vector was constructed. SMA cell model was established by human mesenchymal stem cells(hMSCs) that the vector was transfected into were differentiated to neuron like cells (NLCs).At the same time the control groups were established that the shRNA-0 vector was transfected into and no vector was transfected into. The expression of fl-SMN and delta7-SMN mRNA was observed by RT-PCR analysis. The expression of fl-SMN protein was detected by Western blot.@*RESULTS@#The cells of all the groups were neuron like cells after being differentiated and the protein expression of NSE and NF was positive. The expression of fl-SMN and delta7-SMN mRNA and protein of NLCs in each group was upregulated (P0.05).@*CONCLUSION@#The NLCs, which recombinant SMN1 shRNA expression vector was transfected into, can be regarded as SMA cell model.

Clinical features and prognosis of cerebral venous thrombosis / 中南大学学报(医学版)

OBJECTIVE@#To explore the diagnosis,therapy and prognosis of cerebral venous thrombosis (CVT).@*METHODS@#Twenty-two CVT patients were reviewed. The onset age, clinical manifestations, imaging, treatment, and prognosis were analyzed.@*RESULTS@#Their age ranged from 15 to 58 (mean 33.0+/-8.8) years. Nine were males and 13 were females (1:1.4), 41% of whom were women of childbearing age.This disease occurred rapidly, and the relative pathogeny could be found in most patients (59%), and the hypercoagulative state was the commonest one.The clinical manifestations were variable. Most patients had symptoms and signs of intracranial hypertension(86%), accompanied with or without focal neurological dysfunction and seizures. Disorders of consciousness were found in some sever conditions.The cerebrospinal fluid (CSF) pressure was significantly increased, and the quantity of proteins or white blood cells in CSF was nearly normal.The occluded dural sinus and the clot could be visualised directly by means of magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) or digital subtraction angiogrophy (DSA).After dehydration,anticoagulation,application of adrenal cortex hormone and etilogical treatment,9 patients improved,7 nearly cured, 2 had no changes,1 had cerebral hemorrhage, and 3 died.@*CONCLUSION@#CVT should be suspected when patients show manifestation of intracranial hypertension and/or focal neurological dysfunction and seizures. MRI and MRA are efficient choices for the early diagnosis of CVT. Early diagnosis and anticoagulation with heparin are keys to good prognosis.

An experimental research on differentiation of mesenchymal stem cells derived from children with spinal muscular atrophy into neuron-like cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Spinal muscular atrophy (SMA) is an autosomal recessive neurodegenerative disease. It is characterized by selective loss of spinal cord motor neurons leading to muscle atrophy and is the result of mutation or deletion of the survival motor neuron (SMN) gene. Currently, there are no effective therapies for this disease. Stem cell therapy is a new prospect for SMA patients. This study aimed to investigate whether mesenchymal stem cells (MSCs) can be differentiated into neuron-like cells (NLCs) in SMA patients in order to provide a basis for stem cell therapy for SMA.</p><p><b>METHODS</b>SMA was definitively diagnosed using polymerase chain reaction-restriction fragment length polymerphhism (PCR-RFLP). Two children without SMN1 gene deletion were used as controls. MSCs were isolated and purified from SMA patients and controls, and induced into NLCs by bFGF and baicalin. The NLCs were identified by immunofluourescence staining with NSE and NF monoclonal antibodies.</p><p><b>RESULTS</b>SMA patients showed the deletion of SMN1 exon 7. The morphous and proliferative speed of MSCs between SMA patients and controls were similar. After 6-day induction, MSCs of the two groups displayed similar morphology to that of neurons, with long processes forming extensive networks. NSE and NF, the neuronal markers, were detected in the differentiated NLCs of the two groups.</p><p><b>CONCLUSIONS</b>SMN1 deletion appears not to affect the proliferation and differentiation of MSCs. MSCs of SMA patients can be differentiated into NLCs.</p>

Relationship between single nucleotide polymorphisms of paraoxonase 2 and stroke / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the relationship between single nucleotide polymorphisms of paraoxonase 2 (PON2) and stroke.</p><p><b>METHODS</b>Objects examined comprised of three groups: 120 healthy people, 150 patients with cerebral hemorrhage, 180 patients with cerebral infarction. The PON2 genotypes were determined with PCR and digested by specific restriction enzymes.</p><p><b>RESULTS</b>C311S and G148A polymorphisms of PON2 gene existed among population of Chinese Hunan area, with the allele frequencies 0.23/0.77 for C/S and 0.57/0.43 for G/A in the control group. There was no significant difference of genotype and allele frequency between stroke patients and controls (P>0.05).</p><p><b>CONCLUSION</b>C311S polymorphism of PON2 has no significant correlation with stroke in Han people of Chinese Hunan area and allele C/S is not an independent risk factor for stroke,neither is G148A.</p>

Lymphoplasmapheresis for Guillain-Barre syndrome / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the clinical therapeutic effect and security of lymphoplasmapheresis (LPE) for Guillain-Barre syndrome (GBS).@*METHODS@#Sixty-six GBS patients were randomly divided into 2 groups: the therapy group (33 patients) were treated with LPE in addition to the medical treatment; the control group (33 patients) only accepted the medical treatment. The therapeutic effect was evaluated with the initial recovery time of myodynamia and the myodynamia score difference, and the side effect of the therapy group was observed.@*RESULTS@#The therapy group were treated with LPE for 48 times,1.5 times per person. The average initial recovery time was quicker in the therapy group compared with that in the control group [(6.45+/-3.01) vs (8.36+/-3.83) days]. The difference of limb myodynamia score between pre-treatment and post-treatment in the therapy group was more than that in the control group. The improved number in the therapy group was more than that in the control group, but the ineffective number in the therapy group was not as many as that in the control group. The total effective rate in the therapy group was higher than that in the control group (51.5% vs 27.7%); the average hospital day in the therapy group was shorter than that in the control group [(19.42+/-7.25) vs (24.00+/-8.64) days]; and the difference had statistical significance(P0.05). After the second and the third LPE, the average myodynamia score continued to rise, and the difference had statistical significance (P<0.05). The incidence rate of side effects in the therapy group was 12.5%. Urticaria and hypotension were the major side effects, but they were light and could be relieved by symptomatic treatment.@*CONCLUSION@#The therapeutic effect of LPE is definite, and the side effect is scarce. LPE is safe and effective, and it is worth of generalization and applying in clinical practice.

Lecithin-cholesterol acyltransferase gene 608C/T polymorphism associated with atherosclerotic cerebral infarction / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the distribution of lecithin-cholesterol acyltransferase gene (LCAT) 608C/T polymorphism in Chinese Han population and the relationship of the polymorphism association with the occurrence of atherosclerotic cerebral infarction.</p><p><b>METHODS</b>The lecithin:cholesterol acyltransferase gene 608C/T polymorphism is identified by polymerase chain reaction (PCR), single-strand conformation polymorphism (SSCP)and restriction fragment length polymorphism (RFLP) in 150 patients with ACI and 122 healthy controls matching age and sex.</p><p><b>RESULTS</b>The distribution of LCAT 608C/T gene polymorphism was in accordance with Hardy-Weinberg equilibrium. The CT genotype frequency (14.0%) and T allele frequency (7.0%) in ACI group were significantly higher than those in control group (P<0.05). The concentration of high density lipoprotein cholesterol (HDL-C) in 608CC subgroups were significantly higher than those in 608CT subgroups both in ACI group and in control group (P<0.05).</p><p><b>CONCLUSION</b>The LCAT 608C/T polymorphism is possibly a predisposing factor in ACI happening of Chinese Han population. T allele frequency is possibly concerned with the metabolism of HDL-C.</p>

Quantitative analysis of SMN gene copies in spinal muscular atrophy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the genetic basis in the patients with clinical diagnosis of spinal muscular atrophy(SMA) but without survival motor neuron telomeric copy (SMN-T) deletion; the relationship between the SMN-C (centromeric) copies and the phenotype; and the distribution of SMN-C and SMN-T copies in the SMA patients, the carriers and the controls.</p><p><b>METHODS</b>Quantitative PCR analysis of SMN-T and SMN-C copies were carried out in 45 patients, 25 consanguineous and 33 control individuals. The patients were identified by clinical manifestation and muscular pathology. Two internal standards of SMN-T and cystic fibrosis transmembrane conductance regulator (CFTR) were constructed. Nonradioactive and nonfluorescence-labelling competitive PCR were used. The numbers of SMN-T and SMN-C copies were determined by calculating the ratios of SMN-T/CFTR and SMN-C/CFTR.</p><p><b>RESULTS</b>Quantitation of SMN-T gene copies in SMA patients revealed that nine cases of type I-III were homozygously deleted. Two cases of type III had only one copy and four cases of type III had two copies. SMA IV and other type cases had two copies. Nine cases of consanguineous individuals had one copy, but other 16 had two copies. All of the normal individuals had two copies. Analysis of SMN-C copies showed that SMA I had < or = 2 copies, II-III had < or = 3 copies, SMA IV and others had 0-3 copies, the consanguineous individuals and normal individuals had 0-3 copies.</p><p><b>CONCLUSION</b>The number of copies determined by PCR quantitative assay of SMN-T is in accordance with the result of PCR qualitative assay of homozygous deletion. Quantitative assay of the number of copies can find out the cases and the carriers of heterozygous deletion. The SMA phenotype is related to the number of copies of SMN-C; the smaller the number of copies the patient has, the severer the patient's phenotype will be. The pathogenesis of SMA IV and other types of SMA may not relate to SMN gene.</p>

Preliminary linkage analysis of a Chinese family with benign familial infantile convulsion / 中华儿科杂志

<p><b>OBJECTIVE</b>Benign familial infantile convulsions (BFIC) is a recently recognized autosomal dominant inherited disorder. This epileptic syndrome typically begins between 3 and 12 months of age with clusters of partial seizures in most cases and carries a good prognosis. So far, three loci have been linked to chromosome 19q12.1-13.1, chromosome 2q24 and chromosome 16p12-q12. The authors performed linkage analysis on this pedigree.</p><p><b>METHODS</b>A four-generation Chinese family was investigated. The total number of members was 32 in this family and two neurologists in Xiangya Hospital gave systemic physical examinations and interictal neurological examinations to nineteen members of this family. Venous blood samples were taken for genetic analysis. DNA was extracted from peripheral blood leukocytes using phenol-chloroform method. Seventeen microsatellite markers spanning the critical regions on chromosomes 19q12-13.1, 2q24, and 16p12-q12 were genotyped. These markers included D19S49, D19S250, D19S414, D19S416 and D19S245 for the 19q region, D2S2380, D2S399, D2S111, D2S2195, D2S2330 and D2S2345 for the 2q region, D16S401, D16S3131, D16S3093, D16S517, D16S3120 and D16S415 for the 16p-q region. The DNA from each sample was amplified for the 17 markers. After polymerase chain reactions (PCR), PCR products of chromosome 19 with markers D19S49, D19S250, D19S414, D19S416 and D19S245 were subjected to electrophoresis on 8% denatured polyacrylamide gel for at least 2 hours and 20 minutes. Then the length of the PCR products was judged in the Strategene Eagle Eye II automated gel image analyzer. For the markers from chromosome 2 and 16, PCR products were scanned at ABI 377 autosequencer. The data of PCR products were analyzed using the software Genescan v3.1, Genetyper v2.1 (Applied Biosystem, CA. USA) and GenoDB v1.0. After Mendelian checking, the eligible genotyping data were used for linkage analysis. LOD scores were calculated by using MLINK program of LINKAGE v5.1, under an assumption of autosomal dominant inheritance and the estimated penetrance was 0.9. The allele frequencies of each marker were assumed to be equal and the disease-allele frequencies were designated to be 1/10,000. The LOD scores were calculated at combination rate (theta) 0.0, 0.1, 0.2, 0.3, and 0.4.</p><p><b>RESULTS</b>Among the 17 selected microsatellite markers, which cover the previously reported regions, seven markers' data (D16S3131, D16S517, D16S3120, D16S3093, D2S2380, D19S250 and D19S414) were omitted due to failed genotyping, low genetic heterogeneity, or failure to pass Mendelian checking. Omission of these markers was to ensure the reliability of our raw data. The two-point LOD scores were below zero for all the markers and the maximum LOD scores at theta = 0.0 were less than -2 for markers D19S49, D19S416, D19S245, D16S401, D16S415, D2S399, D2S111, D2S2195, D2S2330 and D2S2345. Thus, the linkage result showed no evidence that the disease locus is linked to any of these selected markers, which excludes the previously reported candidate regions found in other ethnic families.</p><p><b>CONCLUSION</b>There is no evidence that this Chinese family was linked to one of the following loci: 19q12.1-13.1, 16p12-q12 and 2q24. The results indicated that BFIC showed genetic heterogeneity and the Chinese BFIC families might be mapped on another new locus.</p>

The gene expression patterns of genetic P77PMC epilepsy-prone rats' hippocampus / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To find out the differentially expressed genes in the hippocampus of the rats with genetic epilepsy so as to lay a foundation for exploring the pathogenesis of epilepsy by means of cDNA array technology.</p><p><b>METHODS</b>Gene expression patterns in the hippocampus of the genetic epilepsy-prone P77PMC rats and normal Wistar rats were established using the alpha-32P-labeled cDNA probes hybridized with the Atlas Rat cDNA Expression Array, and then were analyzed by an image analysis instrument to get the differentially expressed genes.</p><p><b>RESULTS</b>Fifteen genes were found having differential expression patterns in hippocampus between the P77PMC rats and the Wistar rats, while there may be many other differentially expressed genes left undiscovered due to having no appropriate image analysis software. And among the fifteen genes, the expression levels of twelve genes in the P77PMC rats were higher than those in the Wistar rats, while the expression levels of the other three genes were lower. The results of reverse transcription-polymerase chain reaction(RT-PCR) have demonstrated the reliability of cDNA arrays method.</p><p><b>CONCLUSION</b>cDNA array is a powerful tool for identifying differential expression genes of epilepsy on large scales. There are several differentially expressed genes in hippocampus of the P77PMC rats and the Wistar rats. All these identified genes could play potentially important roles in the pathogenesis of epilepsy.</p>

Advance in researches on survival of motor neurons protein and its interaction proteins / 中华医学遗传学杂志

Survival of motor neurons(SMN) protein is the product of spinal muscular atrophy(SMA) gene. Now the function researching of SMN protein has become hotspot field to discuss the pathogenic mechanism of SMA. The construction, distribution and function of SMN protein are reviewed in this paper.

Clinical observation on the efficacy enhancing and toxicity attenuating effect of nuzhen yangyin granule to the anti-parkinsonism therapy mainly with Medopa / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe and assess the efficacy enhancing and toxicity attenuating effect of Nuzhen Yangyin Granule (NYG) to the anti-parkinsonism (paralysis agitans) therapy with Medopa and Artane.</p><p><b>METHODS</b>Adopting the randomized double-blinded method, the effect of adding NYG to 30 patients with Parkinsonism in the treated group, who already received anti-Parkinsonism treatment but showing decreased response to Medopa and Artane and with obvious adverse reaction, was observed and controlled by 30 patients treated by adding placebo.</p><p><b>RESULTS</b>The total effective rate in the treated group and the control group was 86.7% and 56.7% respectively, the total syndrome improving rate was 90% and 56.7% respectively and the toxicity attenuating rate 90% and 43.3% respectively, comparison between the two groups showed significant difference (P < 0.05 or P < 0.01). NYG also showed markedly effective in reducing the adverse reactions of Medopa and Artane on digestive, neuro-psychiatric and cardiovascular system.</p><p><b>CONCLUSION</b>NYG has obvious efficacy enhancing and toxicity attenuating effects caused by the anti-Parkinsonism treatment with Medopa and Artane.</p>