Objective:To clone human PRKCB1 gene open reading frame(ORF) in order to further research on it's function.Methods:RT-nested PCR method was adopted to the amplify the total length of human PRKCB1 ORF from human umbilical vein endothelial cell(HUVEC).T vector was inserted into the harvested fragment after a tail was added.With blue white screening,the gene ORF encoding human PKC?Ⅱ was obtained by special primers amplifying recombinant plasmids,and linked into T vector.Then the plasmid was identified by sequencing.Results:The human PRKCB1 ORF was amplified successfully with RT-nested PCR and T/A cloning,and the gene sequence was completely consistent with that reported in GenBank.Conclusion:Human PRKCB1 gene ORF was successfully cloned.The strategy of cloning may provide technical references for some genes hard to be cloned.

Objective: To observe the influencing factors for efficiency of liposome-mediated transfection into human umbilical vein endothelial cell(HUVEC). Methods: Transferring HUVECs by distinct conditions,such as ratios of liposome and plasmid,densities of per well and different times of incubation,and transfection rates were observed and calculated by fluorescent microscope and flow cytometry. Results:⑴ When density of per well exceeded 2?l04 and times exceeded 4 hours,liposome-mediated transfection efficiency of endothelial cells decreased. ⑵With adding plasmid quality, transfection rates increased;while plasmid quality exceeded 1.0 ?g,in condition of 2?l04 per well and liposome volume 2 ?l, transfection efficiency reached the peak level. ⑶With adding liposome volume, transfection rates increased;while liposome volume was 8 ?l,in condition of 2?104 per well and plasmid quality 1 ?g, transfection efficiency was depressed.When ratios of liposome and plasmid were 1:6～1:8,the optimal transfection efficiency was 18.62%. Conclusion: Using optimal transfection parameter,we obtained the optimal transfection efficiency .

Objective To construct the eukaryotic expression vector of human PRKCB1 containing enhanced green fluorescence protein gene and transfer into human umbilical vein endothelial cells(HUVECs),then identify activity of expression protein.Methods The pReceiver-M29-PRKCB1 eukaryotic expression plasmid was constructed by frame amplified from pMD18-T-PRKCB1 plasmid.Then the recombinant plasmids were identified by enzyme analysis and DNA sequencing.According to optimized conditions,the eukaryotic expression plasmids were transfered into HUVECs and observed under fluorescence microscope.After that,transfection efficiency was calculated under random vision.The plasma membrane/cytosol ratio of fluorescence was calculated under confocal microscope.The translocation was identified.Results The gene sequence was completely consistent with that reported in GenBank.The enhanced green fluorescence protein was observed in HUVECs after 48 hours.Transfection efficiency was 18.6%?1.6%.The translocation was observed.Conclusion The eukaryotic expression plasmid is successfullyconstructed and transfered into HUVECs,the translocation was identified.It is a potential tool for screening HUVECs stably expressing human protein kinase C ?2 and isolating protein complex.

Diversified clinical practice mode will be explored preliminarily in population of Eight-year MD in military medical university.The successful teaching includes ideological education with military character,standard and systemic clinical traning,liberal arts education and emphasizing medical consciousness.

Objective To investigate the clinical features of moyamoya disease in children and the prognosis of encephaloduroarteriosynangiosis ( EDAS) . Methods According to the age of first operated patients,317 children with moyamoya disease who received EDAS from January 2004 to December 2010 were divided into 3 groups:infant group (n=16,<3 years of age),preschool group (n=42,3 to 6 years of age),and adolescent group (n=259,6 to 17 years of age). The clinical data and the efficacy of operation of the patients were analyzed retrospectively. Results (1) Among the 3 groups of patients,the incidences of cerebral infarction in the infant group (81. 2%,13/16) or the preschool group (69. 0%,29/42) before procedure were significantly higher than the adolescent group (48. 3%,125/259). There were significant differences (χ2 =11. 741,P<0. 01). (2) Before surgical intervention,the infarct volume enlargement or the recurrence of infarction rate at different parts of brain in the infant group (62. 5%,10/16) was higher than that of the preschool group (31. 0%,13/42) and adolescent group (3. 9%,10/259). There was significant difference (χ2 =77. 437,P <0. 01). (3) The overall rate of favourable prognosis was 86. 4% (274/317). There were significant differences between the 3 groups (χ2 =9. 026,P<0.02). Conclusion The conditions of children with moyamoya disease progresses rapidly and their clinical prognosis is poor. It is safe and effective to perform EDAS early moyamoya disease in children.

Objective Toinvestigatetheclinicalfeaturesandsurgicalprognosisofmoyamoya syndromeinchildren.Methods Theclinicaldataof12childrenwithmoyamoyasyndromeadmittedto the 307th Hospital of People′s Liberation Army from December 2002 to October 2013 were analyzed retrospectively. Eleven of them underwent encephalo-duro-arterio-synangiosis (EDAS). A total of 550 children with moyamoya disease in the same period were used as a control group. The clinical characteristics and surgical efficacy of the children with moyamoya syndrome were summarized and concluded by comparing the clinical data of the two groups,including sex,age of onset,initial symptom,progress symptoms, Suzukiinstallments,imagingfeatures,andsurgicalefficacy.Results Themaleandfemaleratioof the children with moyamoya syndrome was 1∶2. Their mean age of onset was 12 ± 5 years old. There were significant differences in the initial symptom (cerebral infarction and cerebral hemorrhage )and disease progress between the children with moyamoya syndrome group and the control group (5/12 vs. 14. 5%[80/550], 3/12 vs. 61. 8%[340/550],and 5/12 vs. 8.7%[48/550],respectively;all P<0. 05). Within the follow-up period,of the 11 children underwent EDAS,7 cases had no further attack,and 4 cases were improved significantly. There was significant difference in the modified Rankin scale (mRS)between the beforeandaftersurgery(0[0,1]vs.2[1,2];P<0.05).Conclusions Theclinicalfeaturesofthe children with moyamoya syndrome have some differences with those with moyamoya disease. Timely and effective EDAS treatment may effectively prevent disease progression and improve the prognosis of patients.

Objective To investigate the clinical features of familial moyamoya disease in China. Methods The patients w ith familial moyamoya disease admitted to the department of Neurosurgery, the 307th Hospital of PLA from March 2009 to June 2012 w ere analyzed retrospectively. Results Of 1 108 patients w ith moyamoya disease admitted to the department of Neurosurgery, the 307th Hospital of PLA, 87 patients (7.8%) w ith familial moyamoya disease w ere identified. Familial moyamoya disease w as observed both in the Han nationality and the minority nationalities. The male to female ratio w as 1∶1.02. The age at first onset ranged from 8 months to 59 years. There w ere tw o peak ages -of-onset, 5-9 years and 30-34 years, respectively). The most common initial symptom w as cerebral ischemia (74 .7%). The first degree relatives w ere the most affected in patients w ith familial moyamoya disease (78/87, 89 .66%), in w hich the siblings accounted for most of the disease ( 38/78, 48.72%), and the proportions of mother-to-child inheritance (21/78, 26.92%) and father-to-child inheritance ( 19/78, 24.36%) w ere similar. Conclusions There are tw o peak ages-of-onset, cerebral ischemia is the most common initial symptom, the first degree relatives are the most affected in patients w ith familial moyamoya disease in China.

Moyamoya disease is a chronic progressive cerebrovascular occlusive disease of unknow n etiology. Transcranial Doppler sonography (TCD) is a non -invasive ultrasonic examination that can detect the hemodynamic characteristics of intracranial large vessels. At present, there are more applications in screen, auxiliary diagnosis, and surgical outcome assessment in moyamoya disease. This article review s the current applications of TCD in moyamoya disease.

Headache is one of the most common symptoms in children w ith moyamoya disease. Its related pathogenesis, clinical manifestations, and operation effect remain unclear. This article review s the advances in headache in children w ith moyamoya disease by review ing the related literatures on the study of headache in children w ith moyamoya disease.