[Expression of cation-chloride cotransporters KCC2 and NKCC1 in brainstem of para- chlorophenylalanine-induced acute insomnia rats].

OBJECTIVE: To explore the possible roles of KCC2 and NKCC1 in the pathological mechanism of acute insomnia in rats. METHODS: A total of 18 Sprague-Dawley rats were randomly selected into model, interference and normal control groups.The expressions of KCC2 and NKCC1 in brainstem were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.The concentration of intracellular Cl(-) ([Cl(-)]i) in brainstem was detected by fluorescence probe MQAE with laser confocal microscopy. RESULTS: (1) Comparing with the control group, both KCC2 mRNA and protein expression were down-regulated in the model and interference groups (mRNA:0.196 ± 0.021 vs 0.939 ± 0.109, P < 0.05; 0.485 ± 0.026 vs 0.939 ± 0.109, P < 0.05; protein expression:0.363 ± 0.058 vs 0.967 ± 0.155, P < 0.05; 0.663 ± 0.106 vs 0.967 ± 0.155, P < 0.05).However they became up-regulated in the interference group versus the model group (mRNA: 0.485 ± 0.026 vs 0.196 ± 0.021, P < 0.05; protein expression:0.663 ± 0.106 vs 0.363 ± 0.058, P < 0.05). (2) Comparing with the control group, both NKCC1 mRNA and protein expression in the model group were slightly up-regulated.But statistical difference was insignificant (mRNA: 0.344 ± 0.026 vs 0.320 ± 0.019, P > 0.05; protein expression:0.244 ± 0.010 vs 0.230 ± 0.021, P > 0.05).There was down-regulation in the interference group versus the model and control groups (mRNA: 0.066 ± 0.031 vs 0.320 ± 0.019, P < 0.05; 0.066 ± 0.031 vs 0.344 ± 0.026, P < 0.05; protein expression:0.131 ± 0.012 vs 0.230 ± 0.021, P < 0.05; 0.131 ± 0.012 vs 0.244 ± 0.010, P < 0.05). (3) Comparing with the control group, [Cl(-)]i became up-regulated in the model group (0.0315 ± 0.0039 vs 0.0164 ± 0.0019, P < 0.05).It was down-regulated in the interference group versus the model group (0.0182 ± 0.0013 vs 0.0315 ± 0.0039, P < 0.05), but higher than control group without statistical difference (0.0182 ± 0.0013 vs 0.0164 ± 0.0019, P > 0.05). CONCLUSION: The down-regulation of KCC2 and rise of [Cl(-)]i in brainstem may participate in the pathological mechanism of acute insomnia in rats. And the mechanism of sedative-hypnotic diazepam may be operate through an up-regulation of KCC2, a down-regulation of NKCC1 and decreased [Cl(-)]i.

[Neuroprotective mechanism of hydrogen sulfide after cerebral ischemia-reperfusion in rats].

OBJECTIVE: To explore the neuroprotective mechanism of exogenous hydrogen sulfide after cerebral ischemia-reperfusion (I/R) in rats. METHODS: A total of 240 male Sprague-Dawley rats were randomly divided into 4 groups of sham-operation group (I), I/R model group (II), low-dose sodium hydrosulfide (NaHS) group (III) and high-dose NaHS group (IV) (n = 60 each). Reversible middle cerebral artery occlusion (MCAO) model was established by intraluminal suture. However, the rats of group I were operated without a blockage of middle cerebral artery. The occlusions in other groups were removed after 2 h. And at 20 min pre-removal, normal saline, 50 µmol/kg NaHS, and 100 µmol/kg NaHS were injected into the abdomens of rats in groups II, III and IV respectively. At 6 h, 24 h, 48 h and 7 d post-reperfusion, behavioral test was performed to evaluate the neurological deficiency. And triphenyltetrazolium chloride (TTC) staining was used to observe the volume of cerebral infarction. In addition, the protein expressions of tumor necrosis factor-alpha (TNF-α) and heat shock protein 20 (HSP20) in ischemic cortex were measured by immunohistochemistry and Western blot. RESULTS: Compared with I/R group, the neurological deficiency scores and volume of cerebral infarction of groups III and IV significantly decreased. Furthermore, the protein expression of HSP20 was markedly up-regulated in the groups III and IV while there was a marked down-regulation of TNF-α after reperfusion. CONCLUSION: NaHS may exert anti-necrotic and anti-inflammatory function by inducing the expression of HSP20, inhibiting the expression of TNF-α and reducing the size of cerebral infarction so as to protect neurons after I/R injury.

[Clinical and imaging features of adult anoxic-ischemic encephalopathy after cardiopulmonary resuscitation].

OBJECTIVE: To explore the clinical and imaging features of anoxic-ischemic encephalopathy (AIE) patients after cardiopulmonary resuscitation. METHODS: A total of 28 qualified AIE patients during the last decade from Xiangya Hospital, Central South University were recruited and analyzed retrospectively. RESULTS: The symptoms of status epilepticus, acute posthypoxic myoclonus, Lance-Adams syndrome, subarachnoid hemorrhage and cognitive deficits were observed. The abnormal findings of magnetic resonance imaging (MRI) or computed tomography (CT), involving neocortex, basal ganglia and para-ventricular white matter, were also recorded. During the early phase of disease, swollen cortex was present on MRI/CT. However, encephalatrophy appeared during the late phase. The more severe symptoms were observed, the more foci were present on MRI/CT. CONCLUSION: The etiologies of AIE patients are heterogeneous after cardiopulmonary resuscitation. The clinical symptoms and imaging studies are of prognostic significance.

The burden of headache in China: validation of diagnostic questionnaire for a population-based survey.

The objective of this study was to test the validity, in the Chinese population, of the Lifting The Burden diagnostic questionnaire for the purpose of a population-based survey of the burden of headache in China. From all regions of China, a population-based sample of 417 respondents had completed the structured questionnaire in a door-to-door survey conducted by neurologists from local hospitals calling unannounced. They were contacted for re-interview by telephone by headache specialists who were unaware of the questionnaire diagnoses. A screening question ascertained whether headache had occurred in the last year. If they had, the specialists applied their expertise and ICHD-II diagnostic criteria to make independent diagnoses which, as the gold standard, were later compared with the questionnaire diagnoses. There were 18 refusals; 399 interviews were conducted in 202 women and 197 men aged 18-65 years (mean age 44.4±12.6 years). In comparison to the specialists' diagnoses, the sensitivity, specificity, positive predictive value, negative predictive value and Cohen's kappa (95% CI) of the questionnaire for the diagnosis of migraine were 0.83, 0.99, 0.83, 0.99 and 0.82 (0.71-0.93), respectively; for the diagnosis of tension-type headache (TTH), they were 0.51, 0.99, 0.86, 0.92 and 0.59 (0.46-0.72), respectively. In conclusion, the questionnaire was accurate and reliable in diagnosing migraine (agreement level excellent), less so, but adequate, for TTH (sensitivity relatively low, false negative rate relatively high and agreement level fair to good). The non-specific features of TTH do not lend themselves well to diagnosis by questionnaire.

[The clinical and genetic studies in a family of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy].

OBJECTIVE: To investigate the clinical features and genetic diagnostic method of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). METHODS: 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. RESULTS: 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. CONCLUSION: The clinical and molecular genetic features of the family accord with CADASIL.

[Serum levels of pro-inflammatory cytokines in diabetic patients with peripheral neuropathic pain and the correlation among them].

OBJECTIVE: To investigate the change of serum levels of the pro-inflammatory cytokines: macrophage migration inhibition factor (MIF), tumor necrosis factor-alpha (TNF-alpha), and interleukin 6 (IL-6), in patients with diabetic peripheral neuropathic pain (DPNP) and their role in the pathogenesis of related diseases. METHODS: Peripheral blood samples were collected from 28 patients with diabetes mellitus (DM) without complications, and gender ratio- and age-matched 32 patients with diabetic peripheral neuropathy (DPN), 28 patients with DPNP, and 28 normal controls. Dual-antibody ELISA was used to detect the serum MIF, TNF-alpha, and IL-6 levels. RESULTS: The serum MIF, TNF-alpha, and IL-6 levels of the DM, DPN, and DPNP groups were all significantly higher than those of the control group (P < 0. 001, P < 0.05). The serum MIF and TNF-alpha levels of the DPN and DPNP groups were all significantly higher than those of the DM group (P < 0.05 or P < 0.001), while there were not significant differences in the serum MIF and TNF-alpha levels between the DPN and DPNP groups (both P > 0.05). There was not significant difference in the serum IL-6 level among the DM, DPN, and DPNP groups as well (all P > 0.05). The correlation analysis show that the levels of MIF, TNF-alpha and IL-6 each other were positively correlated (r = 0.337, P < 0.01; r = 0.216, P < 0.001; r = 0.281, P < 0.05). CONCLUSIONS: The serum pro-inflammatory cytokines MIF, TNF-alpha, and IL-6 levels play an important role in the pathogenesis of DM and DPN, but may not play an important role in the development of DPNP.

Study of the relationship between gene polymorphisms of paraoxonase 2 and stroke in a Chinese population.

BACKGROUND AND PURPOSE: The paraoxonase (PON) gene family has been demonstrated to be capable of preventing lipid peroxidation and could consequently exert antiatherosclerotic effects. Alteration of enzyme activity due to polymorphisms in the PON genes may influence the development of atheroma and thus affect stroke risk. PON2, the second member of this family, has two polymorphisms, C311S and G148A. Our study explores the relationship between the two polymorphisms of PON2 and stroke. METHODS: 120 healthy individuals, 150 patients with primary cerebral hemorrhage and 180 patients with cerebral infarction were included in this study. The genotypes of PON2 were detected with polymerase chain reaction and digested by specific restriction enzymes. RESULTS: C311S and G148A polymorphisms of PON2 exist in Chinese people, with the allele frequencies 0.273/0.727 for C/S and 0.567/0.433 for G/A. No significant differences were observed in genotype and allele frequency between stroke patients and controls (p > 0.05). CONCLUSIONS: There was no significant correlation between the two tested polymorphisms of PON2 and stroke in the Chinese population. Allele C/S and G/A might not be independent risk factors for stroke.

[Relationship between P-type Na+ -K+ -Atpase and migraine: an experimental study with rats].

OBJECTIVE: To investigate the role of P-type Na+ -K+ -ATPase in the mechanism of migraine. METHODS: nitroglycerin induced migraine Twenty Sprague-Dawley rats, 10 male and 10 female, were randomly divided into 2 groups: model group, undergoing subcutaneous injection of nitroglycerin 10 mg/kg once a week for 4 weeks so as to establish migraine model, and control group, undergoing subcutaneous injection of normal saline. Then the rats were killed with their trigeminal ganglia, trigeminocervical complex, and cortex of frontal lobe taken out. RT-PCR and Western-blotting were used to detect the mRNA and protein expression of P-type Na+ -K+ -ATPase. Immuno-precipitation assay was conducted to observe the Na+ -K+ -ATPase activity. The concentration of intracellular Ca2+ was investigated by Fluo-3/AM fluorometric method. RESULTS: The mRNA and protein expression levels and activity of P-type Na+ -K+ -ATPase in the trigeminal ganglion and trigeminocervical complex of the model group were 0.72 +/- 0.05, 0.59 +/- 0.05, and 5.21 +/- 0.51 respectively, and the mRNA and protein expression levels and activity of P-type Na+ -K+ -ATPase in the cortex of frontal lobe of the model group were 0.70 +/- 0.05, 0.60 +/- 0.05, and 3.61 +/- 0.49 respectively, all significantly lower than those of the control group (0.83 +/- 0.10, 0.67 +/- 0.06, 6.53 +/- 0.73, 0.81 +/- 0.08, 0.71 +/- 0.09, and 6.61 +/- 0.73 respectively, all P < 0.05). The concentration of intracellular Ca2 in trigeminal ganglion and trigeminocervical complex and the concentration of Ca2+ in the cortex of frontal lobe of the model group were 211,182 +/- 12,973 and 186,511 +/- 18,297 respectively, both significantly higher than those of the control group (135,243 +/- 18,105 and 143,289 25,175 respectively, both P < 0.01). CONCLUSION: P-type Na+ -K+ -ATPase may infect the pathogenesis of migraine by its expression and activity.

[Establishment of spinal muscular atrophy cell model by RNAi].

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 (P<0.05), but the expression of delta7-SMN mRNA and protein in SMA model group was lower than that in the control group (P<0.05). The expression of delta7-SMN mRNA between the groups had no statistical difference (P>0.05). 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].

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.

[Relationship between single nucleotide polymorphisms of paraoxonase 2 and stroke].

OBJECTIVE: To study the relationship between single nucleotide polymorphisms of paraoxonase 2 (PON2) and stroke. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

[The clinical diagnosis and therapy of neurosyphilis].

OBJECTIVE: To study the clinical features, neuroimages, laboratory findings, therapy and prognosis of neurosyphilis. METHODS: The clinical features, neuroimages, laboratory findings and therapy of 18 patients with neurosyphilis were analyzed retrospectively. RESULTS: Interstitial neurosyphilis especially meningovascular syphilis was the most common type of which the clinical features varied widely. Treponema pallidum hemagglutination assay (TPHA) test of both serum and CSF samples might be positive, but the positive rate in CSF was lower than that in serum. MRI of syphilitic gumma was typical. Penicillin is the drug of first choice for effective treatment except syphilitic gumma, which should be treated with resectopn. The effective rate of penicillin is 93%. CONCLUSIONS: Patients with neurosyphilis may present diverse clinical symptoms at the beginning of the course, therefore it is misdiagnosed easily at the first visit. It is important to comprehend that positive syphilis antibody test in serum or CSF is specific for the diagnosis. Penicillin is the drug of first choice for effective treatment.

[Effects of short hair RNA on the expression of SMN1 gene in human mesenchymal stem cells].

OBJECTIVE: To construct a plasmid expressing short hairpin RNA (shRNA) targeting SMN1 gene in mammalian cells and to investigate the effect of shRNA on the expression of SMN1 gene in human mesenchymal stem cells (hMSCs), thus providing experimental basis for the establishment of SMA cell model. METHODS: Five shRNAs targeting SMN1 gene was designed according to the SMN1 cDNA sequence provided by GenBank, and cloned to the downstream of U6.1 promoter in the pRNAT-U6.1/Neo vector, thus forming different recombinant plasmids: pshRNA-SMN1, pshRNA-SMN2, pshRNA-SMN3, pshRNA-SMN4, and pshRNA-SMN5 expressing specific shRNAs targeting SMN1 gene. The plasmid psh-RNA-0 was used as controls. The recombinant plasmid pshRNA-SMN1 was transfected into human mesenchymal stem cells (hMSCs) by Lipofectomine (TM) 2000, and its effects on the mRNA expression and protein expression of fl-SMN were observed by RT-PCR and Western blotting. RESULTS: The successful construction of recombinant plasmid pshRNA-SMN was identified by enzyme cutting and DNA sequencing. After transfection, the expression levels of fl-SMN mRNA and protein in hMSCs of the pshRNA-SMN1-1 group and pshRNA-SMN1-4 group were significantly lower than that of the control group (both P < 0.05) with an interfering efficiency of 64.05% and 61.04% in mRNA level, and 52.97% and 61.57% in protein level respectively. However, the expression of Delta7-SMN mRNA was not effected. CONCLUSION: The constructed recombinant plasmid called pshRNA-SMN1 inhibits the expression of fl-SMN mRNA and protein in hMSCs efficiently.

[Lymphoplasmapheresis for Guillain-Barre syndrome].

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(P<0.05). The average myodynamia score after the first LPE increased, but the difference had no statistical significance (P>0.05). 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.

[An experimental research on differentiation of mesenchymal stem cells derived from children with spinal muscular atrophy into neuron-like cells].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: SMN1 deletion appears not to affect the proliferation and differentiation of MSCs. MSCs of SMA patients can be differentiated into NLCs.

Involvement of BDNF/TrkB and ERK/CREB axes in nitroglycerin-induced rat migraine and effects of estrogen on these signals in the migraine.

Migraine is a highly prevalent headache disorder, especially in women. Brain-derived neurotrophic factor (BDNF) and its receptor tropomyosin receptor kinases (TrkB), as well as extracellular signal-regulated kinase (ERK) and its downstream target c-AMP-responsive element binding protein (CREB) are strongly associated with the transmission of nociceptive information. However, the involvement of these substances in migraine has rarely been examined. In the present study, intraperitoneal injection of nitroglycerin (NTC) successfully induced rat migraine attack, as evidenced by behavioral testing. The location and abundance of these substances in the migraine model were determined by immunohistochemistry, real-time polymerase chain reaction (RT-PCR), western blot and enzyme-linked immunosorbant assays (ELISA). Results showed that BDNF, TrkB, phosphor(p)-ERK and p-CREB were up-regulated in the brain neurons of both male and female rats with NTG-induced migraine compared to non-migraine control, whereas their expression levels were decreased in headache-free intervals of the migraine compared to migraine attacks. Estrogen is an important contributor to migraine. Female ovariectomized rats showed significant reduction in the expression of BDNF, TrkB, p-CREB and p-ERK in both attacks and intervals of NTG-induced migraine, relative to rats that have their ovaries. But, intraperitoneal administration of exogenous estrogen recovered their expression in ovariectomized rats. Collectively, this study unveiled a positive correlation of BDNF/TrkB and ERK/CREB axes in NTG-induced migraine and promoting effects of estrogen on their signals in the migraine. These findings contribute to further understanding the pathogenesis of migraine in the molecular basis.

[Lecithin-cholesterol acyltransferase gene 608C/T polymorphism associated with atherosclerotic cerebral infarction].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

[The expression of SMN2 gene mRNA in neuron-like cells derived from patients with spinal muscular atrophy].

OBJECTIVE: To detect the difference of SMN2 mRNA expression between the neuron-like cells (NLCs) derived from patients with spinal muscular atrophy (SMA). METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to diagnose the patients with SMA and the controls. Mesenchymal stem cells (MSCs) were induced into neuron-like cells which become the models of neurons. The transcripts of SMN2 were testified with RT-PCR combined with sequencing. RESULTS: Two bands (266 bp and 212 bp) were found in the gel picture of RT-PCR and the band of 266 bp was the full length transcript (fl-SMN mRNA). The band of 212 bp was testified by sequencing to be deletion of the exon 7, which was the production of alternative splicing (skipping exon 7, SMNDelta7 mRNA). The expression of fl-SMN mRNA accounted for 23.2% of the total SMN mRNA in the SMA patients, being much lower than the rate in the controls (82.0%). In contrast, 76.8% of SMN gene transcripts was SMNDelta7 mRNA in the SMA patients, being much higher than the rate of 18% in the controls. CONCLUSIONS: Alternative splicing exists in SMN2 gene transcription, that is, exon 7 is skipped during the processing of SMN2 mRNA in the NLCs of SMA patients, which is one of the reasons of the full-length SMN protein lacking.

[Human mesenchymal stem cells differentiate into neuron-like cells and show SMN protein expression].

OBJECTIVES: To investigate the mRNA and protein expression of SMN gene in neuron-like cells by inducing MSC to differentiate into neuron-like cells. METHODS: Human MSC (hMSC) were isolated and purified. Human MSC were treated with basic fibroblast growth factor (bFGF) before inducing hMSC to differentiate into neuron-like cells with dimethylsulfoxide (DMSO) and butylated hydroxyanisole (BHA). The expression of NSE, NF, and SMN protein in neuron-like cells were detected by immunohistochemistry. The expression of SMN mRNA were detected with reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Human MSC progressively assumed neuronal morphological characteristics after being induced for 3 hours. Cell bodies had long process contacting with each other. The neuronal marker of NSE and NF was positive in neuron-like cells. Both hMSC and neuron-like cells expressed the mRNA of SMN gene. Compared with hMSC the latter expressed highly the mRNA of SMN gene and it was significance (P < 0.01). SMN protein was only expressed in neuron-like cells. CONCLUSIONS: Human MSC are able to differentiate into neuron-like cells with expressing the neuronal marker. Neuron-like cells can express highly the mRNA of SMN gene and express SMN protein.