BACKGROUND: Myasthenia gravis (MG) patients with thymoma were often neglected in clinical work and delayed the therapy.OBJECTIVE: To investigate the significance of the Ryanodine receptor (RyR) antibody on the assessment of MG.DESIGN: A case analysis.SETTING: Institute of neurology in a hospital of a university.PARTICIPANTS: This experiment was carried out in the Institute of Neurology, Fudan University from June 1999 to March 2002. There were 66 MG patients with thymoma(MGT group), 98 MG patients with non-thymoma (NTMG group), 50 patients with non-myasthenia gravis(NMG) and 123 normal persons (NC group).METHODS: Sarcoplasmic reticulum(SR) abounded in RyR was extracted with differential centrifugation, in order to establish a detecting system of ELISA-RyR-RyR antibody (RyR-ab).MAIN OUTCOME MEASURES: The levels of RyR-ab in serum of researched subjects.RESULTS: Positive rate of RyR-ab in MGT group was higher than that in NTMG and NMG groups(P ＜ 0.01), moreover, the sensitivity and the specificity were 81.8% and 94.5% respectively. The positive rates of MGT groups with different thymus histology were no significant difference(P＞ 0.05). Ages, clinical scores and levels of acetylcholine receptor antibody (AchR-ab) in patients with positive RyR-ab were higher than those in patients with negative RyR-ab( P ＜ 0.01 ) in MG group. The levels of RyR-ab was positive correlated with the severities of clinical symptoms in MG patients, especially the patients in MGT group( r = 0. 626, P ＜ 0.01) . And among the different histological types of MGT, thymoma of epithelioid cells has the highest correlation coefficient ( r = 0. 592, P ＜ 0. 01).CONCLUSION: The detection of RyR-ab has better sensitivity and specificity for the diagnosis of MGT and the levels of RyR-ab is positive correlatied with the severities of MG patients.

Objective To explore the neuroprotective mechanisms of granulocyte colony stimulating factor (G-CSF) in ischemic brain injury. Methods Brain tissues were taken out from MCAO and sham operated Sprague-Dawley rats 7 days after operation. The expression of G-CSFR, GDNF, MAP2 and GFAP was measured by using immunofluorescence co-staining. Results The expression of G-CSFR and GDNF were widely distributed in the neurons in normal brain tissues, not in the astrocytes. However, in ischemic peripheral zone, part of G-CSFR and GDNF positive cells merged with GFAP positive cells, in normal brain tissues, most G-CSFR positive cells were co-expressed with GDNF. Conclusion Cerebral ischemia induces astrocytes to express G-CSFR and GDNF, suggesting that endogenous neuroprotection by cerebral ischemia may be related with the expression and production of G-CSFR and GDNF in astrocytes in ischemic peripheral zone.

Objective To investigate the role of anti-herpes ximplex virus (HSV)-1 IgM secreting cells detection assay in early diagnosis of herpes simplex encephalitis. Methods Twenty-three herpes simplex encephalitis cases and 40 control cases were included in this study. Anti-HSV-1 IgM secreting cells and anti-HSV-1 IgM were retrospectively tested in the patients' cerebrospinal fluid by enzyme-linked immunosorbent spot (ELISPOT) and enzyme-linked immunosorbent assay (ELISA), respectively. The data analysis was performed by using Fisher Exact Test. Results Using ELISPOT method for detection of 9 HSV-1 encephalitis patients' and 16 clinical control cases anti-HSV-1 IgM secreting cells within two weeks after disease onset, the sensitivity of ELISPOT for detecting anti HSV-1 IgM secreting cells in the cerebrospinal fluid was 88.9% (8/9) and the specificity was 93.8%(15/16). On the other hand, the sensitivity of ELISA for detecting anti-HSV-1 IgM in ccrebrospinal fluid was 16.6% (2/12) and the specificity was 88.2% (15/17) when using ELISA method for detection of 12 HSV-1 encephalitis patients' and 17 clinical control cases's anti-HSV-1 IgM secreting cells. The sensitivities of the two methods were statistically different (P<0.01). Conclusion Compared to ELISA, ELISPOT for detecting the anti-HSV-1 IgM secreting cells in cerehrospinal fluid is a more sensitive method for early diagnosing herpes simplex encephalitis.

Objective To survey gene expression profiles in nonlesional refractory temporal lobe epilepsy(TLE)and to further verify the difference of gene expression.thus to evaluate the possible molecular pathogenesis of this kind of epilepsy that can help to supply a new way for the diagnosis and treatment.Methods The TLE samples and control cases were studied by means of cDNA microarray consisting of 1 8 000 genes.Reverse transcription polymerase chain reaction(RT-PCR)Was performed to measure the expression alterations of SH3GL2.BTNN2A2 and KCNJ4 mRNA in temporal cortex samples from patients who had undergone temporal lobectomy surgery for intractable epilepsy.Tissue from 10 subjects who did not have epilepsy served as controls.Results The known genes differently expressed in those TLE samples involved immunity correlation factor genes,signal conduction genes,ion channel transportation genes;mitochondria function genes and SO on were identified.Among which.the expression of SH3GL2 mRNA Was significantly increased in epileptic brain(1.022±0.547)compared with the controls(0.446±0.171,t=-3.181).In TLE group(0.481±0.196),the expression of BTN2A2 mRNA was also significantly higher than that of control subjects(0.243±0.111,t=3.351).Compared with control group(O.795±0.112),the expression of KCNJ4 mRNA Was significantly decreased in TLE patients(0.438±0.178).Conclusions cDNA microarray is an efficient and high.throughout method to survey gene expression profiles in intractable temporal lobe epilepsy.The variation of those gene expressions might be a potential etiological agent for TLE that may offer a novel target for anticonvulsant therapy.

Objective To establish an early diagnostic test for tuberculous meningitis (TBM) with good sensitivity and specificity. Methods Twenty-five patients with a clinical diagnosis of TBM and 49 controls, including 27 patients with other infectious diseases of central nervous system and 22 patients with noninfectious neurological diseases, were enrolled in our research. We simultaneously detected antimycobacterium bovis BCG IgG secreting cells in both cerebral spinal fluid (CSF) and peripheral blood (PBL)by enzyme linked immunospot assay(ELISPOT),repeated insertion sequence IS6110 specific for mycobacterium tuberculosis in CSF by PCR and anti-BCG IgG titre in both CSF and PBL by enzyme linked immunosorbent assay(ELISA).Results The sensitivity of ELISPOT was 84.0%,much higher than that of PCR(75.0%)and ELISA(52.3%).The specificities of the three tests were 91.8%,93.7%and 91.8%respectively.The numbers of CSF cells secreting anti-BCG IgG tested by ELISPOT were even higher in the early phase of TBM, but declined along with the disease progressing(t=-3.183,P=0.008),which allowed an early diagnosis to be made. Conclusion ELISPOT technique is proved to be the most valuable test for the early diagnosis of TBM.

Objective To investigate the correlation between survival motor neuron (SMN) gene deletion and Chinese patients with sporadic amyotrophic lateral sclerosis (SALS).Methods A total of 141SALS patients and 134 unrelated controls were recruited from the Chinese population.Polymerase chain reaction (PCR) and restriction fragment length polymorphisro (RFLP) analysis were performed to screen SMN gene deletion.Frequencies of deletion were coropared by Chi-square test.Results Four patients and 3 controls were detected to have horoozygous SMN2 deletion.The frequencies of SMN2 deletion were 2.84%(4/141) and 2.24% (3/134), respectively, which was not significantly different (χ2= 0.0001, P =1.000).No subjects were found to have homozygous SMN1 deletion.Condusion There is no correlation between SMN gene deletion and Chinese patients with SALS.

OBJECTIVETo understand the possible pathogenesis of sporadic multiple system atrophy (MSA).

METHODSThe immunoreactivity and ultrastructural features of glial cytoplasmic inclusions (GCIs) in 12 autopsy patients with MSA and 4 normal control groups were studied. All regional sections from each subject were evaluated with HE staining, Klüver-Barrera (KB), Holzer's, modified Gallyas-Braak's (GB) methods and immunohistochemical staining with alpha-synuclein and ubiquitin antibodies. Pontine white matter with abundant GCIs from case 1 was examined, using conventional electron microscopy, Gallyas-Braak's electron microscopy and immunoelectron microscopy.

RESULTSThe presence of GCIs as constantly demonstrated in all MSA patients. Strong alpha-synuclein immunoreactivity was observed in all of the ubiquitinated GCIs. However, the density of alpha-synuclein positive GCIs differed from case to case, and there was no relationship between the density of GCIs and age, sex, or MSA subtype. Ultrastructural features indicated that argyrophilic granule-associated filaments of about 25 nm in diameter were the predominant constituents of GCIs, and the anti alpha-synuclein antibody selectively labeled in these filaments. No GCIs and alpha-synuclein immunoreaction were found in control brain tissues.

CONCLUSIONSGCI was a pathognomonic change in sporadic MSA patients. Accumulation of alpha-synuclein in GCIs may occur during the early stags of MSA. Seletcive alpha-synuclein positive abnormal microtubules in GCIs therefore play an important role in the pathogenesis of MSA.

Aged ; Aged, 80 and over ; Female ; Humans ; Immunohistochemistry ; Inclusion Bodies ; ultrastructure ; Male ; Microscopy, Immunoelectron ; Middle Aged ; Multiple System Atrophy ; etiology ; metabolism ; pathology ; Nerve Tissue Proteins ; analysis ; Neuroglia ; ultrastructure ; Synucleins ; alpha-Synuclein

Objective To investigate the clinical and pathological features of Uurich congenital muscular dystrophy (UCMD). Methods The clinical aspects of 3 patients with UCMD, 2 with Duchenne muscular dystrophy (DMD) and 1 with congenital muscular dystrophy 1A (MDC1A) were analyzed. And the muscle specimens from these patients were studied using immunohistochemistry and immunofluorescence staining. Results UCMD was clinically characterized by neonatal hypotonia with proximal contracturos and distal hyperlaxity at birth or early infancy. Histochemical staining revealed muscle frber hypoplasia andinterstitium proliferation. Immunohistochemistry staining with anti-collagen Ⅵ antibody revealed complete(1/3) or partial (2/3) deficiency of collagen Ⅵ in the sarcolemma and interstitial matrix. Partial deficiency was better demonstrated by immunofluorescence staining. Conclusions The proximal contractures and distal hyperlaxity is the clinical hallmark of UCMD. Collagen Ⅵ immunolabelling can confirm the diagnosis of UCMD.