ObjectiveTo observe the effect of early rehabilitation on patients with severe closed traumatic brain injury in intensive care unite (ICU).Methods118 patients with severe closed traumatic brain injury were divided randomly into the treatment group and control group with 59 cases in each group. The patients of control group recieved respiration rehabilitation, bedsore prevention and urinary system nursing; while the patients of treatment group added with motor and cognitive rehabilitation. Scores of Glasgow coma scale (GCS), Fugl-Mayer assessment (FMA), mini-mental status examination (MMSE), disability rating scale (DRS) and activities of daily living (ADL) of all patients were recorded before and after treatment.ResultsThe scores of GCS, FMA (upper limbs), ADL and DRS of the treatment group patients were significantly difference with that of the control group after treatment (P<0.05). The MMSE scores of two groups had no significantly difference after treatment (P>0.05).ConclusionEarly rehabilitation on the patients of severe closed traumatic brain injury in ICU can improve brain function and reduce the disability rating scale. The improvement of cognitive ability is not obviously in a shot period.

Objective: To establish a method using non-ionic detergent for extracting lipid rafts. Methods: Because lipid rafts can resist solubilization by non-ionic detergents under 4℃, we use non-ionic detergent (Triton X-100) to treat with epicyte fractions, the non-raft membrane would be solubilized. Then we utilize sucrose gradient centrifugation, preparations enriched in lipid rafts could be obtained.caveolin-1 was used as markers of lipid-raft structures. Results:A white light-scattering band under light illumination located at the interface between 15%-20% sucrose was detectable, and a brown stripe which comparative molecular quantity is 24 000 was identified by Western-Blot analysis. Conclusion: The method using non-ionic detergent is simple and useful for extracting lipid rafts, extracting lipid rafts would be prerequisite in studying the function of lipid rafts.

In this study we implanted magnetically labeled neural stem cells (NSCs) in PD rats and then monitored their survival and migration in the host brain by magnetic resonance imaging (MRI). The mesencephalic NSCs were obtained from the brain of SD rats. Superparamagnetic iron oxide (SPIO) was transferred to NSCs by Lipofectamine transfection. Eighteen PD lesioned rats were selected for transplantation by evaluation of their rotational behavior in response to amphetamine and randomly assigned to 3 groups, i.e., sham group, PBS group and NSCs transplanted group, with 6 rats in each group. MR scanning was performed at 1, 2, 4, 6, 8 and 10 week(s) following transplantation. At the meantime, rotational behavior was assessed in each group. Our results showed that SPIO particles were clearly visible with Prissian blue staining in neurospheres and cells derived from NSCs. The rotational behavior of the NSCs transplanted group was remarkably improved compared with that of sham group and PBS group (P < 0.05). In vivo MR tracking of NSCs showed that SPIO labeling led to a strong susceptibility change of signal 1 week after transplantation on T2 weighted images. And a large circular hypointense signal appeared in the transplanted area on T2* gradient echo images. Ten weeks following transplantation, the hypointense signal on T2 weighted and T2* gradient echo images was still displayed. It is concluded that SPIO particles could label NSCs effectively, and MRI detection of SPIO labeled cells is a promising method and novel approach to analyzing the NSCs following transplantation in the treatment of PD.

AIM: To study effect of endogenous carbon monoxide on intracellular calcium concentration and explore the mechanism in brain protection of endogenous CO in focal cerebral ischemia in rats. METHODS: SD rats were divided into three groups randomly, which including hemin, ZnPP group and saline group as control. Respectively saline, hemin, ZnPP were injected intra-peritoneally twelve hours before middle cerebral artery was occluded. Twenty four hours after MCAO model was set up, the concentration of carbon monoxide in blood and intracellular calcium in neural cells was examined. RESULTS: Contrast to saline group, the concentration of CO in blood rose up while intracellular calcium in occluded side decreased in hemin group; the concentration of CO in blood went down while intracellular calcium in occluded side rose up in ZnPP group, there was significant difference among them (P0.05). CONCLUSIONS: It may be one of mechanisms on brain protection in ischemic cerebral tissue that carbon monoxide affected intracellular calcium concentration of neural cells by regulating Ca~(2+)-K~+ channel on cell membrane as a messenger gaseous molecular and neurotransmitter. [

ObjectiveTo study the anatomical abnormalities of basal ganglia and research their influence on depression status in patients with post stroke depression (PSD)by diffusion tensor imaging (DTI) of MRI.MethodsPatients with basal ganglia infarction were recruited,and divided into groups of PSD and non depression control group by Hamilton Depression Rating Scale (HAMD) assessment. All the patients were evaluated with National Institute of Health Stroke Scale ( NIHSS). And the patients were checked by DTI sequence.Fractional anisotropy (FA),average diffusion coefficient (ADC) values and the number of nerve fiber were measured in bilateral caudatum,pallidum,putamen and thalamus.ResultsThe score of NIHSS (6.29 ± 3.45 ) was significantly higher in PSD group than that in non-depression group (3.95 ± 1.90 ;t =2.219,P =0.036). No significant difference was found between the two groups for the DTI data of the basal ganglia nuclei ( t =0.056-1.618,all P ＞ 0.05 ). Compared with contralateral construction (0.40 ± 0.02 ),the FA value decreased in the left putamen ( 0.37 ± 0.03 ) in the PSD group ( t =2.243,P =0.045 ).By Spearman correlations analysis,the HAMD score was positively correlated with NIHSS score ( r =0.464,P =0.017 ),and negatively correlated with the FA values of left pallidum (r=-0.563,P=0.005),right pallidum (r=-0.416,P=0.035) and left putamen (r =-0.428,P =0.029).Conclusions The occurrence of PSD was associated with neurological functional deficit following basal ganglia infarction.The depression level was correlated with the increasing of NIHSS score,the reductions in bilateral pallidum and left putamen FA values.This research contributes to evaluation of the PSD status in patients with basal ganglia infarction.

Objective To explore the effect of extracellular regulated protein kinases (ERK) signaling pathway on early brain injury and autophagy of nerve cell in hippocampus area in rats with subarachnoid hemorrhage (SAH). Methods Forty-eight adult male Sprague-Daw-ley rats were randomly divided into sham group, SAH group, SAH+dimethyl sulfoxide (DMSO) group and SAH+U0126 group, with 12 rats in each group. The SAH model was established with puncture of internal carotid artery. The SAH+U0126 group was injected with U0126 0.05 mg/kg;the sham group and SAH group were injected with normal saline, and the SAH+DMSO group was injected with DMSO 30 min-utes before modeling. They were sacrificed 24 hours after modeling. The brain water content was measured with wet and dry method. The morphology changes of neural cells in hippocampus CA1 were observed by HE staining. The expression of phosphorylation ERK (p-ERK), Beclin-1 and LC3-Ⅱwere detected with immunohistochemical method and Western blotting. Results Compared with the sham group, the brain water content increased (P<0.05), the number of survival neurons decreased (P<0.05), the expression of p-ERK, Beclin-1 and LC3-Ⅱincreased in SAH group (P<0.05). Compared with SAH group, the brain water content increased, the number of survival neurons decreased (P<0.05), the expression of p-ERK, Beclin-1 and LC3-Ⅱ decreased in SAH+U0126 group (P<0.05); and no significant difference was found in SAH+DMSO group (P>0.05). Conclusion The activation of ERK signaling pathway may alleviate early brain injury after SAH by regulation of autophagy.

ABSTRACT:Objective To explore role of U0126,the specific inhibitor of ERK signaling pathway,in early brain injury (EBI)and the autophagy of nerve cells in hippocampus area in subarachnoid hemorrhage (SAH). Methods A total of 48 male adult SD rats were randomly divided into control group,SAH group,DMSO+SAH group,and U0126+SAH group,with 12 in each.We established SAH rat model by the puncture of internal carotid artery.The same amount of saline water,DMSO and U0126 solution of 0.5 mL per rat was injected respectively into the rats of different groups 30 min before modeling.The rats were killed at 24 h.To measure brain water content by Wet and dry method after 24 h,the morphological changes of hippocampus CA1 neural cells were observed by microscopy;the expression levels of ERK,Beclin-1 and LC3 were detected by using immunohistochemical method. Results Compared with that in sham group,brain water content increased obviously in SAH model group.The density of surviving neurons in SAH group was significantly lower than that in control group (P<0 .0 5 ).ERK signaling pathway was activated obviously,the expressions of Beclin 1 and LC3-Ⅱ were significantly higher than those in control group (P<0.05).Compared with SAH model group,in U0126 group brain water content increased obviously.Compared with those in SAH group,the density of surviving neurons was significantly lower (P<0.05), ERK signaling pathway was suppressed,the expressions of Beclin-1 and LC3-Ⅱ were significantly lower (P<0.05). Conclusion The U0126,the ERK signaling pathway inhibitor,can inhibit neuron autophagy and increase EBR of SAH.

In this study we implanted magnetically labeled neural stem cells (NSCs) in PD rats and then monitored their survival and migration in the host brain by magnetic resonance imaging (MRI).The mesencephalic NSCs were obtained from the brain of SD rats. Superparamagnetic iron oxide (SPIO) was transferred to NSCs by Lipofectamine transfection. Eighteen PD lesioned rats were selected for transplantation by evaluation of their rotational behavior in response to amphetamine and randomly assigned to 3 groups, i.e., sham group, PBS group and NSCs transplanted group, with 6 rats in each group. MR scanning was performed at 1, 2, 4, 6, 8 and 10 week(s) following transplantation.At the meantime, rotational behavior was assessed in each group. Our results showed that SPIO particles were clearly visible with Prissian blue staining in neurospheres and cells derived from NSCs.The rotational behavior of the NSCs transplanted group was remarkably improved compared with that of sham group and PBS group (P＜0.05). In vivo MR tracking of NSCs showed that SPIO labeling led to a strong susceptibility change of signal 1 week after transplantation on T2 weighted images.And a large circular hypointense signal appeared in the transplanted area on T2* gradient echo images.Ten weeks following transplantation, the hypointense signal on T2 weighted and T2* gradient echo images was still displayed. It is concluded that SPIO particles could label NSCs effectively, and MRI detection of SPIO labeled cells is a promising method and novel approach to analyzing the NSCs following transplantation in the treatment of PD.

OBJECTIVE: To assess the value of chromosomal microarray analysis (CMA) and next-generation sequencing (NGS) for the analysis of abortic tissues. METHODS: A total of 242 samples of spontaneous abortion were collected and tested by CMA or NGS. RESULTS: The detection was successfully in 238 cases (98.35%). In total 143 cases of chromosomal abnormalities were detected, which accounted for 60.08% of all cases. Numerical chromosomal abnormalities were found in 133 cases(93.01%), structural abnormalities were found in 9 cases (6.29%), and uniparental disomy was found in 1 case(0.70%). CONCLUSION Both CMA and NGS have the advantages of high-throughput, good coverage, high resolution and rapid analysis. They can be used for the detection of the causes of spontaneous abortions. CMA is more useful for the detection of aneuploidies and uniparental disomy, while NGS has advantages in its throughput, capacity in detecting low percentage chimerism and cost, which can provide more options for clinicians.
Abortion, Spontaneous ; genetics ; Chromosome Aberrations ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Microarray Analysis ; Pregnancy

Objective To explore the mutation characteristics of DMD gene in patients with Duchenne or Becker muscular dystrophy and female carriers, to provide effective prenatal diagnosis. Methods Samples were collected from 94 male patients clinically diagnosed with Duchenne or Becker muscular dystrophy and 121 corresponding female relatives from Qingdao Women and Children′s Hospital from June 2011 to October 2018. Multiplex ligation-dependent probe amplification (MLPA) was used to detect their DMD gene, and 23 high risk pregnants were performed prenatal diagnosis. Any candidate of DMD gene single-exon deletion was validated by further PCR amplification. The sample with whole DMD gene deletion was confirmed by chromosomal microarray analysis (CMA) to detect copy number variations and break site. Results Among 94 clinical Duchenne or Becker muscular dystrophy patients, 66(70.2%, 66/94) were detected gene mutation; 56 cases were exon deletion mutation and 10 cases were duplication mutation. In 121 female relatives, 48 cases (39.7%, 48/121) were diagnosed as carriers. The mutation carrying rate, was 64.5% (40/62) identified in 62 mothers of Duchenne or Becker muscular dystrophy patients. Five Duchenne or Becker muscular dystrophy fetuses and 5 carrier fetuses were prenatally diagnosed in 23 high risk pregnants. Two children with the entire DMD gene deletion were identified more deletions at Xp21, with deletions of 6.66 Mb and 10.64 Mb respectively. Conclusions MLPA may be an important method to detect DMD gene mutation of deletion and duplication. Therefore, the diagnosis of probands, female carriers and making an effective prenatal diagnosis are essential to reduce the birth of children with Duchenne or Becker muscular dystrophy.