Brain ; pathology ; Caudate Nucleus ; pathology ; Child, Preschool ; Humans ; Huntingtin Protein ; Huntington Disease ; diagnosis ; genetics ; pathology ; Magnetic Resonance Imaging ; Male ; Nerve Tissue Proteins ; genetics ; Pedigree ; Trinucleotide Repeats ; genetics

The occurrence of epilepstic seizures in patients with cerebral palsy (CP) remains a serious event and is very disruptive for children who already suffered from orther disabilties. Data from population-based studies or neurodevelopmental clinic studies showed that 8%～62% of patients with CP suffer from epilepsy. The incidence is highest in spastic tetraplegia and hemiplegia. Half of the patients were onset within the first year of age. Children with tetraplegia CP tended to have an earlier onset of epilepsy than chlidren with other CP types. Partial seizures were the most common seizure types, followed by infantile spasms and Lennox-Gastaut syndrome. Low birth weight, neonatal seizures, family history of epilepsy, lower intelligence and grey matter damage were found to be related to significantly increased risk of epilepsy. Epilepsy in children wih CP usually had poor outcome, half of them were intractable and needed polytherapy. Only 37%～65.2% of patients became seizure-free.

OBJECTIVE: To establish an HPLC-MS method for the analysis of the impurity profile of cefotaxime sodium. METHODS: Shimadzu-LCMS-IT-TOF was used with Waters XBridge Shield (RP18, 4.6 mm×250 mm, 5 μm) column. Mobile phase A was 20 mmol·L-1 ammonium acetate (pH adjusted to 6.25)-methanol (92: 8), and mobile phase B was set at 20 mmol·L-1 ammonium acetate-methanol (60: 40) (pH adjusted to 6.25).Gradient elution was performed at a flow rate of 1.0 mL·min-1. ESI source was used.Positive and negative ion scanning was conducted in the range of m/z 150-900.The heating temperature was 200℃, CDL temperature was maintained at 200℃, atomization gas flow rate was 1.5 L·min-1, dry gas pressure was 94.0 kPa, and the post-column diversion ratio was 1: 4.Some related substances in cefotaxime sodium were identified by comparing the retention time in chromatography, [M+H]+ spectrum and MS2 spectrum with those of reference substances, the others which haven't reference substances were deduced or speculated by analyzing the MS2 or MSn fragmentation with the help of a rule summarized from the MS2 fragmentation of cefotaxime sodium and the reference substances of system suitability impurities. RESULTS: Twenty-six related substances were separated and detected in the sample, all of which were identified or deduced. They were cefotaxime sodium isomeric compounds and homologs generated during the production process or degradation products. CONCLUSION: The method can be applied in the identification and qualitative analysis of the related substances of cefotaxime sodium and the quality control and optimization of the synthesis of cefotaxime sodium.

Objective To characterize the language development and articulation in children with different types of cerebral palsy. Methods The data from 76 children with cerebral palsy who underwent standardized tests of speech and language were analyzed. The incidence and abnormal pattern were compared between groups. ResultsLanguage delay and/or dysarthria were noted in 73.1% of these subjects. Children with diplegia developed similar levels both in comprehension and verbal expression, whereas those with tetraplegia and athetoid developed poorer verbal expression. The incidence of language delay in children with diplegia, tetraplegia and athetoid were 45.95%, 90% and 64.7%, respectively. Dysarthria was found in all the children with tetraplegia, athetoid and ataxia, but only 48.65% of those with diplegia. Conclusion Language disorders were common in children with cerebral palsy, especially those with athetoid and tetraplegia.

ObjectiveTo investigate the value of diffusion tensor imaging (DTI) at high b value for unilateral middle cerebral artery (MCA) occlusive disease in patients without obvious infarct lesions on conventional MR imaging.MethodsDTI at high b value (2200 s/mm2 ) was performed using a 3.0 Tesla MR scanner in 34 patients with unilateral middle cerebral artery occlusion,who had no obvious infarct lesions on conventional MR imaging. Fractional anisotropy (FA),apparent diffusion coefficient (ADC),axial diffusivity (eigenvalue λ1) and radial diffusivity (eigenvalues λ2,λ3) were measured at the ipsilateral and contralateral corona radiata,anterior and posterior limbs of the internal capsule,cerebral peduncle and pons in all subjects.Mean ADC,FA,λ1,λ2 and λ3 values of corona radiata,anterior and posterior limbs of the internal capsule,cerebral peduncle and pons were compared between the ipsilateral and contralateral MCAterritory by t test. Results Among the 34 patients,left MCA occlusion in M1 segment occurred in 16 patients and right MCA occlusion in Ml segment occurred in 18 patients.At the ipsilateral corona radiata,mean FA,ADC,λ1,λ2 and λ3 were 0.419 ±0.032,(5.975 ±0.272) × 10 3,(5.704 ±0.365) ×10-3,(6.412 ±0.368) × 10-3 and (6.605 ±0.343) × 10-3 mm2/s,respectively.At the contralateral corona radiata,mean FA,ADC,λ1,λ2 and λ3 were 0.443 ± 0.033,(5.804 ± 0.282) × 10 -3,(5.651 ±0.350) × 10-3,(6.099 ±0.353) × 10-3 and(6.372 ±0.355) × 10-3 mm2/s,respectively.At the ipsilateral corona radiata,mean FA was significantly decreased(t =11.614,P ＜0.01),and mean ADC (t=12.421,P＜0.01),λ1(t =7.447,P＜0.01),λ2(t=10.244,P＜0.01) and λ3(t=9.890,P＜0.01) were significantly increased.At the ipsilateral anterior and posterior limb of the internal capsule,mean FA were 0.609 ±0.026 and 0.674 ±0.033,λ1 were(5.330 ±0.462) × 10 -3 and(5.171 ±0.456) ×10-3 mm2/s,respectively.At the contralateral anterior and posterior limb of the internal capsule,FA were 0.622 ±0.026 and 0.694 ±0.034,λ1 were(5.064 ± 0.448) × 10 -3 and(4.924 ± 0.365) × 10 -3 mm2/s,respectively.Mean FA was significantly decreased (t =7.823,8.013,all P ＜ 0.01) and mean λ1 was significantly increased (t =7.811,8.800,all P ＜0.01) at the ipsilateral anterior and posterior limbs of the internal capsule.There was no significant difference in ADC,λ2 and λ3 value between the ipsilateral and contralateral sides.And all the DTI parameters,including mean ADC,FA,λ1,λ2 and λ3 values,showed no statistical difference between both sides of cerebral peduncle and pons.ConclusionDTI at high b valuc can provide useful information for visualizing ischemic white matter injury in patients without obvious infarct lesions on conventional MR imaging.

Objective To analyze the comorbidities and gross motor function classifications (GMFCs) of premature children with cerebral palsy (CP) in terms of neurological subtype and gestational age in search of some relationship. Methods Children with cerebral palsy treated at the Qingdao Children's Hospital from 2006 to mid2009 received intelligence capacity tests, ophthalmological consultations, language/speech tests, brainstem auditory evoked potential measurements, electroencephalograms and GMFC evaluations. All of the children were stratified according to neurological subtype, gestational age, comorbidities and gross motor function classification. Results Of all 258 children, spastic diplegic cerebral palsy predominated (183 case, 70.9% ). 124 cases (48.1% ) had visual disorders, 121 (46.9%) had language or speech disorders and 103 (39.9%) showed mental retardation. The frequencies of individual comorbidities were distributed disproportionately between the different neurologic subtypes.GMFC levels also differed with the different CP types. The GMFC levels of diplegics were significantly better than those of the other types. The distribution of comorbidities such as visual disorders, language or speech disorders, and mental retardation was not related to gestational age or type of CP. Conclusions There is some correlation between the neurological subtype, comorbidities and the GMFC levels. But there is no significant correlation between gestation age and the severity of CP.

Objective To investigate neuroimaging and its correlation with clinical aspects of cerebral palsy (CP).Methods A retrospective study of 295 children with CP was conducted.Magnetic resonance imaging (MRI),the gross motor function classification system (GMFCS) and intelligence testing were administered,and any correlations among these measures was analysed.Results Among the 295 cases,257 presented abnormal MRIs (87.1％) due to brain maldevelopment (n =11),periventricular leukomalacia (PVL) (n =173),cortical/subcortical lesions (n =17),basal ganglia lesions (n =26),cerebellar maldevelopment (n =11) or others lesions (n =19).Thirty-nine presented with normal MRIs.About 26％ were rated at GMFCS level 1,18％ at level 2,17％ at level 3,19％ at level 4 and 20％ at level 5.Almost 82％ of the children presented with brain maldevelopment and 84.4％ with PVL-induced spastic bilateral paralysis.In 41％ of the children with cortical or subcortical lesions,induced spastic hemiplegia was observed,whereas 47％ had induced spastic bilateral paralysis.In 77％ of the children with basal ganglia lesions induced involuntary movement was observed,and all of those with maldevelopment of the cerebellum were ataxic.Most of those with spastic hemiplegic,bilateral paralysis,involuntary movement and ataxia were on GMFCS levels 1 or 2,with only 3.7％,33.5％,64.1％ and 46.2％ respectively on GMFCS level 4 or 5.Among those in whom the MRI revealed brain maldevelopment,9.1％ were on GMFCS level 1 or 2.The corresponding percentage for PVL was 43.9％,for cortical or subcortical lesions 58.8％,for basal ganglia lesions 19.2％ and for cerebellar maldevelopment 27.3％.The balance in each category were on GMFCS level 4 or 5.Epilepsy was most common in the children with brain maldevelopment (36.4％) or cortical or subcortical lesions (41.2％).Mental retardation was most common in cases of brain maldevelopment (45.5％),cortical or subcortical lesions (41.2％) or cerebellum maldevelopment (36.4％).The incidence of epilepsy and mental retardation was higher among the children on levels 4 and 5 than on levels 1 and 2.Conclusions Neuroimaging correlates significantly with the type of CP and GMFCS level.Epilepsy and mental retardation are most common in children with brain maldevelopment or lesions.The incidence of epilepsy and mental retardation is higher among children rated at GMFCS level 4 or 5 than among those on levels 1and 2.

Objective To analyze the endoscopic features of patients with the lower esophageal sphincter (LES) and crural diaphragm (CD) separation and explore its significance in hiatus hernia (HH) diagnosis.Methods From January 2011 to June 2012,the data of patients underwent esophagus high resolution manometry (HRM) examination were retrospectively analyzed and of which the patients with LES-CD separation were selected.Patients who received digestive tracts operation or lack of complete endoscopic date were excluded.A total of 93 cases were enrolled for analysis.According to Chicago Criteria,the cases were divided into three groups.There were 21 cases in type Ⅰ group (LES-CD separation＜1 cm),37 cases in type Ⅱ group (LES-CD separation≥l cm and ＜2 cm),35 cases in type Ⅲ group (LES-CD separation≥2 cm).The differences among groups in endoscopic features were analyzed.HRM were taken as golden standard,the sensitivity and specificity of endoscopy in HH diagnosis were analyzed.Measurement data were expressed as x ± s,one-way ANOVA was performed for multi-groups comparisons,LSD method or Dunnett T3 test for pairwise comparisons and chi-square test or Fisher's exact test for two groups' comparisons.Results There were no significant differences in the distance between esophagus gastric junction (EGJ) and foreteeth of the three groups (P＞0.05).Distances between EGJ and diaphragmatic HH pressure impression in type Ⅲ group,type Ⅱ group and type Ⅰ group were (3.57±0.78) cm,(1.89±0.81) cm and (1.14±0.67) cm,respectively and the differences were significant (LSDt=9.26,11.44,3.57; all P＜0.05).The percentage of continuously-open cardia and showing pressure impression of the gastric pouch of type Ⅲ group was higher than that of type Ⅱ group and the percentage of continuously-open cardia and showing pressure impression of gastric poach of type Ⅱ group was higher than that of type Ⅰ group (80.0％,40.5％ and 4.8％,x2 =11.64,29.76 and 8.59; 91.4％,27.0％ and 4.8％,x2 =30.69,40.73 and 4.32' all P＜0.05).The occurrence rates of the His angle blunting and sac of type Ⅲ group were higher than those of typeⅡ group and type Ⅰ group (74.3％ and 77.1％,24.3％ and 24.3％,4.8％ and 4.8％,x2 =17.97 and 25.41,both P＜0.05),there was no significant difference between type Ⅱ group and type Ⅰ group (P＞0.05).The rates of esophagitis of type Ⅲ group and type Ⅱ group were higher than that of type Ⅰ group (71.4％,59.5％ and 14.3％,x2 =17.14 and 11.15,both P＜0.05),there was no significant difference between type Ⅲ group and type Ⅱ group (P＞0.05).There was one case of Barrett's esophagus in each group.The sensitivity and specificity of endoscopy in HH diagnosis were 91.4％ and 81.0％,respectively.Conclusions The distance between EGJ and hiatal pressure impression,continuously-open cardia,the His angle blunting,hiatal pressure impression of the gastric pouch and hernia sac were important indicators of HH endoscopic features.The sensitivity of endosocopy is higher than the spcificity of endosocopy in HH diagnosis.

Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.

AIM:To explore the mechanisms underlying contraction induced by extracelluar acidosis (pHex6.8) in rat isolated coronary artery (RCA).METHODS:Using the microvessel tension recorder system, the effects of acid-base transporters on RCA contraction induced by pHex6.8 were explored by applying the selective pharmacological inhibitors of Na+-H+ exchanger 1 (NHE-1) and Na+-HCO-3 cotransporter (NBC), HOE-642 and S0859, respectively.The effects of chloride channel on RCA contraction induced by pHex6.8 were explored by applying the inhibitors of chloride channel (NPPB and NFA), and by replacing the extracellular NaCl with equimolar NaBr.RESULTS:pHex6.8 augmented the resting tension of RCA, and the maximum contraction was (3.90±0.95) mN.HOE-642 at 30 μmol/L and S0859 at 100 μmol/L both inhibited the contraction of RCA induced by pHex6.8 (P<0.01).NPPB and NFA both inhibited the contraction of RCA induced by pHex6.8 or KCl (60 mmol/L) in a concentration-dependent manner.NPPB and NFA (100 μmol/L) both inhibited the contraction of RCA induced by U46619 (1 μmol/L).Replacing the extracellular NaCl with equimolar NaBr almost completely inhibited RCA contraction induced by pHex6.8 (P<0.01), but had no obvious effect on the contraction induced by KCl (60 mmol/L) or U46619 (1 μmol/L).CONCLUSION:Extracellular acidosis-induced contraction in RCA may be related to the activated NHE-1 and NBC, and it may be also related to the enhanced chloride transport across the membrane.