Objective To investigate the expression of aquaporin-4 (AQP4) in cultured astrocytes after in vitro hypoxia induced by CoCl2. Methods After primary culture and subculture, the astrocytes were placed in a controlled atmosphere culture chamber. Both control group and hypoxia groups were established.These groups were further divided into seven sub-groups according to the different time intervals: 15, 30minutes and 1,2, 4, 6, 12 hours, respectively (6 apertures for each group). The shape of the astrocytes in each group was observed with light microscopy and transmission electron microscopy ( TEM ). All groups were examined using in situ hybridization, real time fluorescence quantitative reverse transcriptase polymerase chain reaction, immunocytochemistry and Western blot. The data was analyzed statistically with SPSS 13.0 software. Results There was significant consistency between the AQP4 mRNA and protein ( r =0. 85, P ＜0. 01 ). There was slight positive expression of AQP4 in a few astrocytes of the control groups. In the hypoxia groups, the expression of AQP4 increased within 15 minutes; the increase was most prominent between 1 and 4 hours( mRNA in hypoxia groups: 0. 26 ± 0. 04, 0. 31 ± 0. 02, 0. 36 ± 0. 04; control groups:0. 06 ±0. 01,0. 09 ±0. 01,0. 08 ±0. 01 )after hypoxia and became less between 6 and 12 hours; There was significant difference in the AQP4 expression between the hypoxia groups and control groups among different time points (t = 16. 51, 18.20, 15.26,all P＜0. 01 ). The corresponding pathological changes were cellular edema, which was most prominent between 1 and 4 hours. Under TEM, increase in size of the nucleolus and swelling of endoplasmic reticulum and mitochondria; these changes became more marked with time.Disruption of a few astrocytes was detected in the hypoxia groups at 12 hours. Conclusions The pathological change of astrocytes is cellular edema following hypoxia. There is a positive relationship between the presence and degree of cellular edema as well as the duration of hypoxia and the up-regulating of AQP4.These results imply that AQP4 expression is an important molecular mechanism of celluar edema of astrocytes.

Objective To explore the expression of vascular endothelial growth factor (VEGF)mRNA and protein in endothelia progenitor cells (EPCs) and VEGF in the culture medium and serum of patients during acute period of cerebral hemorrhage associated with hypertension (APCHH). Methods Mononuclear cells from peripheral blood of patients with APCHH ( 16 patients) and hypertension ( 16 patients) were isolated and induced to EPCs. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to assay VEGF mRNA. VEGF protein was assessed by Western blotting. The VEGF protein level in patient's serum and culture medium ( at day 7 ) were assayed using VEGF ELISA Kit and compared between APCHH group and hypertension group. Results Compared with hypertension group, VEGF mRNA (0. 186 ±0. 035 versus 0.090 ±0.031, t =8.318, P ＜0.0l ) and protein (0. 223 ± 0. 028 versus 0.169 ± 0. 022, t = 3. 744, P ＜ 0. 01 ) expression of EPCs, the concentration of VEGF protein in the supernatant (414 ±37 versus 316 ±29, t =8. 270, P ＜0. 01 ) and in serum (408 ±49versus 222 ±34, t = 12.406, P ＜0. 01 ) were all significantly increased in APCHH group. Conclusion The VEGF protein levels in serum of patients and in the culture medium, VEGF mRNA and protein expression in EPCs were all significantly increased during acute periods of cerebral hemorrhage.

Objective To explore the associations between coagulation factor Ⅶ (FⅦ)polymorphisms and its haplotype with risk of ischemic cerebrovascular diseases (ICVD) in Henan Han population. Methods Five hundred and twelve cases with ICVD as patient group and 560 healthy subjects as control were recruited in the study. The polymorphisms of R353Q, 5'F7 and IVS7 were detected by PCR-RFLP. The genotype frequency and allele gene frequency were compared between ICVD group and control group. The haplotype was analyzed by SHEsis software. Results The RQ genotype frequencies and Q allele frequencies of ICVD group were significantly lower than those of control group. The distribution of H7 allele frequencies and H6H7 genotype frequencies of FⅦ/IVS7 polymorphisms had significant difference between ICVD group and control group. Finally, the prevalence of R-P0-H6 haplotype in ICVD group(53. 3% )was higher than that in control group (47.5%, OR = 1. 219, 95% CI 1. 028-1. 446,P =0.023). Conclusions In Henan Han population, the Q allele of F Ⅶ/R353Q polymorphisms and the H7 allele of F Ⅶ/IVS7 polymorphisms may be protective genetic factors against ischemic cerebrovascular disease, and the R-P0-H6 haplotype may be a risk factor of ischemic cerebrovascular disease.

Objective To summarize the clinical and pathological features of glycogen storage disease (GSD) type Ⅱ. Methods The clinical and pathological data of the 20 GSD type Ⅱ patients were reviewed. Results One patient with infantile-onset mainly presented hypotonia, muscle weakness, feeding difficulties, pulmonary infection and cardiomyopathy insufficiency and increase of serum creatine kinase (778 IU/L) and echographic evidence of hypertrophic cardiomyopathy were detected. Electromyography studies indicated a definite myopathy. Nineteen cases were late-onset, presenting a slowly progressive proximal myopathy with truncal involvement or with symptoms dominated by respiratory insufficiency. Not all muscles were equally affected. Increase of serum creatine kinase (208-2600 IU/L) was detected in 14 patients and normal level in 1 patient. Electromyography studies indicated a definite myopathy in 9 patients,with abnormal irritability in 1 patient and susceptible in 4 patients and myotonic discharge in 1 patient and no abnormalities in 2 patients. Echographic evidence of thickening of the interventricular septum and pulmonary hypertension were detected in 2 patients respectively. The common light microscopic feature of all case was a vacuolar myopathy with high glycogen content and acid phosphatase activity in the vacuoles. Conclusions GSD type Ⅱ often presents slowly progressive myopathy which often affect the toro and respiratory muscles.In most patients the serum creatine kinase level is elevated slightly. Muscle biopsy is of use to make the definite diagnosis of this disease.

Objective To evaluate the detection of culture filtrate protein 10 (CFP10) and 6000 early secretory antigenic target (ESAT-6) in cerebrospinal fluid to be used in diagnosing tuberculous meningitis. Methods Dot enzyme linked immunosorbent assay ( Dot ELISA) method that was improved by applying concentrated cerebrospinal fluid was used to detect CFP10 and ESAT-6 in cerebrospinal fluid to analyze small protein antigen secreted by M. tuberculosis. Cerebrospinal fluid of 111 subjects were collected,in which 58 specimens were clinically diagnosed as tuberculous meningitis and 53 as non-tuberculous.CFP10 and ESAT-6 were detected in cerebrospinal fluid using Dot ELISA method and the results were analyzed. Results The sensitivities of detecting CFP10 and ESAT-6 antigen were 93.1% and 91.4% respectively, and the specificities were 92. 5% and 94. 3% respectively. The sensitivities and specificities are generally higher compared with the other methods of detecting M. tuberculosis or materials of M. tuberculosis by acid-fast staining or mycobacterium tuberculosis culture and polymerase chain reaction.Conclusions Using Dot ELISA method to detect CFP10 and ESAT-6 in cerebrospinal fluid to diagnose tuberculous meningitis has a high sensitivity and specificity. Our study provided the evidence of detecting the specific antigen of M. tuberculosis to be used in diagnosing tuberculosis.

Objectives To assess the value of external anal sphincter electromyography (EASEMG) in evaluating the related autonomic dysfunction in Parkinson's disease ( PD), parkinsonism dominant multiple system atrophy (MSA-P) and progressive supranuclear palsy (PSP). Methods From the records of EAS-EMG collected in our lab (total 562 cases), 60 PD (male 41, female 19), 68 MSA-P (male 35,female 33) and 13 PSP (male 10, female 3) were included in the analysis in this study. Mean duration,polyphasic ratio and satellite potential occurrence rate were comparable among the groups. Mean duration prolongation were graded as normal ( ＜ 10.0 ms), mild ( 10.0-11.9 ms), moderate ( 12.0-13.9 ms)and severe ( ≥ 14.0 ms). Results Among all related autonomic symptoms, the occurrence rate of constipation, urinary incontinence, urgency and frequency in patients with MSA-P(95.8% (23/24) ,94.6% (53/56) ,87.7% ( 50/57 ), 85.7% (42/49), 76.5% ( 39/51 ) ) were higher than that of PD ( 61.5%(16/26), 62.3% (33/53), 30.6% (15/49), 46.2% (24/52), 45.7% (21/46)) and PSP (75.0%(3/4) , 62.5% (5/8), 50.0% (4/8), 42.9% (3/7), 42.9% (3/7)). The abnormal rate of EAS-EMG in PD, MSA-P and PSP were 60.0%, 94.2% and 84.6%, accordingly. Mean duration ( PD ( 12.0 ± 1.6)ms, MSA-P (15.4±3.0) ms, PSP (13.8±1.8) ms), polyphasic ratio (PD 46.2% ±19.2%, MSA-P 63.9% ± 15.8%, PSP 51.5% ± 12.1% ) and satellite potential occurrence rate ( PD 9.5% ± 8.3%,MSA-P 26.5% ± 15.9%, PSP 19.2% ± 12.5% ) varied significantly different among the groups ( F =31.724, F = 17.412, x2 =45. 335, all P ＜0.01 ). Severe mean duration prolongation was overwhelming in MSA-P (66.2% ) , compared with mild 10.3% and moderate 23.5%. The predominant prolongation degree was moderate in PSP (61.5%, mild 7.7%, severe 30.8% ), and mild in PD (36.7%, moderate 36.7% ,severe 11.7%, normal 15.0% ). Conclusions EAS-EMG could play a role in evaluating the related autonomic dysfunctions in PD, MSA-P and PSP. The EAS-EMG impairment was severe and frequent in MSA-P, mild and infrequent in PD, moderate in PSP. The spectrum of mean duration prolongation suggested the possibility of Onuf's nucleus involvement in these diseases.

Objective To investigate the clinicopathologic features of the brain tissue from multilobar resection or hemispherectomy for refractory epilepsy. Methods The clinical and pathologic findings of 46 cases seen at Xuanwu hospital from 2005 to 2009 were reviewed retrospectively. Results The mean age of seizure onset and disease duration were 3.9 years and 10.2 years, respectively. There were 33 cases of hemispherectomy and 13 cases of multilobar resection. Temporal lobe abnormality was seen in all cases. The pathologic subgroups were as follows: ulegyria (31/46), malformation of cortical development (MCD, 7/46 ) and infection (8/46). Microscopic examination of ulegyria showed cortical architectural disturbances, neuronal loss, reactive gliosis and appearance of corpora amylacea. We also noted deposition of hemosiderin (13 cases), calcification (9 cases) and island-like neurons (5 cases). All ulegyria cases were accompanied by varying degree of cortical dysplasia, and hippocampus sclerosis were identified in 7 cases. MCD comprised of 5 cases of focal cortical dysplasia ( FCD), including 3 cases of FCDⅠB, 1 case of FCDⅡA and 1 case of FCDⅠA, 1 case of polymicrogyria and 1 case of porencephaly. Among 8 infection eases, there were 5 cases of Rasmussen encephalitis ( RE), l case of cysticercosis, 1 case of tuberculous meningitis and l case of Cytomegalovirus encephalitis. Conclusions The most common pathological category of specimens from hemispherectomy or multilobar resection is ulegyria with obvious temporal lobe abnormality. This is followed by MCD ( with FCDⅠB as the main type) and central nervous system infection (RE as the most frequent abnormality).

Objective The quantitative pharmacoelectroencephalography ( QPEEG ) of many antiepileptic drugs (AEDs), such as carbamazepine, valproic acid, phenobarbital and topiramate but not levetiracetam (LEV), have been studied. This study is to investigate the effect of LEV on QPEEG. Methods One dose of LEV at l g was administrated to 12 healthy adults (6 males, average age at 26 years, average height at l.67 m). The EEG samples (of 180 seconds each) were obtained prior to and at regular intervals ( 1, 1.5, 2, 3, 4, 6, 7, 8, 12, 24 hours) after administration of LEV. The EEG activity was processed with the power spectral analysis and separated into different frequency bands. The absolute powers of both occipital and frontal lobes were calculated through 30 seconds epochs without artifacts for each recording. The statistical difference between baseline pre-drug control and each post-drug assessment was evaluated by the Wilcoxon matched-paired rank test. Results The power of α1-band and β-band increased bilaterally over both frontal and occipital lobes after the administration of LEV. There was no change of α2-band over bilateral frontal lobes, but increased in double peaks shape with the low point at the 6 hours after the administration. The power of α1-band showed the significant change after the administration at the 1.5 hours (18.8950, Z= -3.059, P=0.002) in the left front, 3 hours (18.6150, Z= -2.981, P =0.003)in the right front, 1.5, 2, 3 and 4 hours (61.0233, 53.9425, 47.6192 and 51.8250 respectively, Z =-3.059, -3.059, -2.903 and -3.059, all P ＜ 0.01 ) in the left occipital, and 1, 1.5, 2 and 3 hours (53.5358, 56.8092, 50.3500 and 47.1733 respectively, Z = -2.903, -3.059, -3.059 and -2.981,all P ＜ 0.01 ) in the right occipital. The power of α2-band showed the significant change after the administration at the 3 hours (73.5450, Z = - 3.059, P = 0.002) in the left occipital, and 1, 3 hours (80. 6808 and 87. 1750, Z = -2.903 and - 3.059, P = 0.004 and 0.002 respectively ) in the right occipital. The power of β-band showed the significant change after the administration at the 3 hours (3.8000, Z = -3.059, P = 0.002) in the left front, 1.5, 2 and 3 hours (4.0408, 4.3217 and 4. 1050,Z= -2.903, -3.059 and -3.061, all P＜0.01) in the right frontal, 3 hours (9.1408, Z= -3.059,P =0.002) in the left occipital, and 1.5, 3 hours (8.9267 and 9.3033, Z = -2.981 and -2.981, both P = 0.003) in the right occipital. Conclusions LEV can change the background activity of QPEEG. The changes are different from those of the other AEDs.

Objective To investigate the feature of the morphology changes in the upper airway in patients with acute cerebral infarction and to find a new method to prevent and cure cerebral infarction.Methods Sixty-six patients with cerebral infarction confirmed by brain MRI or CT scan(within 3 weeks of onset) were recruited.The patients were examined by upper airway MRI scan and polysomnography (PSG).Then the patients were divided into obstructive sleep apnea hypopnca syndrome(OSAHS)group and non-OSAHS group.In addition.16 patients showing OSAHS but without stroke history(OSAHS nonstroke group)were included in the study.The sagittal and horizontal lengths of the nasopharynx,palatopharynx,glossopharynx and hypopharynx were measured and their closs-sectional areas were calculated.The length,thickness and cross-sectional area of the palate were also measured.Statistic analysis of each data among the groups was performed using SPSS software.Results Among 66 cases with acute cerebral infarction,75.8 % (50/66)were diagnosed with OSAHS.The anteropesterior diameer,left and right diameters and smallest section area in upper airway were all smaller in the OSAHS group with acute cerebral infaretion than those in the non-OSAHS group and OSAHS non-stroke group.The narrowest segments in upper airway were nasopharynx and ompharynx.which are caused by shortened left and right diameters.The area of the soft palate in the OSAHS-stroke group was significant bigger((452.2±99.6)mm2)than that in non-OSAHS group((350.0±69.4)mm2,t:4.575,P<0.05).The lowest SO2 in OSAHS-stroke group(68.9 % ±10.5 % )was the lowest among three groups.The more severe the airway constriction was.the higher the apnea-hypopnea index(AHI)was and the lower the lowest SO2 was.Conclusion Patients withl stroke show higher incidence of OSAHS and present more severe multilevel upper airway constriction.Upper airway constriction may be the new target of early treatment for better prognosis of cerebral infarction.

Objective To report the clinical and pathological features of the sensory neuropathy caused by a combined therapy of telbivudine and pegylated interferon α-2a in 2 patients with hepatitis B virus infection Methods Two male patients aged 48(case 1)and 20(case 2),who suffered from hepatitis B virus infection.were given telbivudine and pegylated interferon α-2a.After 4 months treatment,both patients developed numbness and pain in the lower limbs.The physical examination showed decreased pain sensation in distal extremities.Hypahidrosis appeared in distal extremities.The nails were pale changed in fingers and toes in cage 1.Case 2 presented mild weakness in the proximal muscle of lower limbs and the tendon reflex was decreased in both lower limbs.His 8erunl creatine kinase level was mild elevated.The electromyography examination and sural nerve biopsies were performed on both patients.Results Electromyography examination showed significant decrease of amplitude of sensory nerve action potentials and mild decrease of sensory nerve conduction velocities in both patients.The amplitude of motor nerve action potentials was also decreased in case 2.Light microscope examination revealed middle reduction of myelinated fibers,wallerian degeneration of myelinated fibers and small clusbers of regenerated fibers in sural nerve.Electro microscopy examination revealed the loss of unmyehnated nerve fibers.After the combined therapy was stopped and vitamin B,CoQ10 and L-camitine were administered,the patients recovered gradually.Conclusions Combined therapy of telbivudine and pegylated interferon α-2a may cause sensory neuropathy with electrophsiological and pathological abnormalities of axonal lesions.The sensory neuropathy induced by the combined therapy may be reversible.