1.Cloning and sequencing analysis of human PKC?Ⅱgene ORF
Journal of Chongqing Medical University 1987;0(01):-
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.
2.The influencing factors for efficiency of liposome-mediated transfection of pReceiver-M29-PRKCB1 into human umbilical vein endothelial cell
Journal of Chongqing Medical University 1987;0(01):-
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 .
4.MicroRNA and hypertension.
Lian DUAN ; Xing-Jiang XIONG ; Jie WANG
China Journal of Chinese Materia Medica 2014;39(3):397-401
MicroRNA(miRNA) is involved in virtually all biologic processes, including cellular proliferation, apoptosis, and differentiation. Thus, miRNA deregulation often results in impaired cellular function and disease development, so miRNAs have potential therapeutic relevance. The elucidation of these processes regulated by miRNAs and the identification of novel miRNA targets in the pathogenesis of hypertension is a highly valuable and exciting strategy that may eventually led to the development of novel treatment approaches for hypertension. Several mechanisms have been implicated in the pathogenesis of hypertension: overactivation of therenin-angiotensin-aldosterone system (RAAS), dysfunction of the vascular endothelium, damnification of vascular smooth muscle. To maintain and restore target organ expression of miRNA stable may be a new strategy for treatment of hypertension. The article reviews pathogenesis of miRNA and hypertension, researches of miRNAs as biomarker and therapeutic target, discusses advances in miRNA-based approaches that may be important in treating hypertension.
Animals
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Biomarkers
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metabolism
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Humans
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Hypertension
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drug therapy
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genetics
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metabolism
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MicroRNAs
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genetics
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metabolism
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Molecular Targeted Therapy
5.Study on glucose, creatinine, urea nitrogen and serum ET-1 of patients with acute cerebral infarction
Xinwang DUAN ; Lian HUANG ; Huihua LI
Chinese Journal of Emergency Medicine 2011;20(10):1089-1091
Objective To study glucose,creatinine,urea nitrogen and serum ET-1 of patients with acute cerebral infarction,and to explore the relationship between neurologic impairment and ET-1 levels.Methods The glucose,creatinine,urea nitrogen and serum ET-1 were retrospectively analyzed in 50 patients with acute cerebral infarction ( < 24 h) and 50 patients with non-neurological diseases.ET-1 determined by 125I radioimmunoassay.Results There were no significant differences in glucose,creatinine and urea nitrogen of acute cerebral infarction ( P > 0.05 ) ; Compared to the control groups,ET-1 levels was significantly higher ( P < 0.01 ),and levels of serum ET-1 in acute cerebral infarction were significantly correlated with their neurological deficits ( P < 0.01 ).Conclusions Levels of serum ET-1 can severd as diagnostic and prognostic indicator of acute cerebral infarction.
6.Clinical features of moyamoya disease in children and the efficacy analysis of encephaloduroarterio-synangiosis
Bing ZHU ; Xiangyang BAO ; Lian DUAN
Chinese Journal of Cerebrovascular Diseases 2014;(6):284-288
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.
7.Clinical features of familial moyamoya disease in China
Zhengshan ZHANG ; Rui ZHANG ; Lian DUAN
International Journal of Cerebrovascular Diseases 2016;24(2):114-116
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.
8.Application of transcranial Doppler sonography in moyamoya disease
Chenfan XIE ; Cong HAN ; Lian DUAN
International Journal of Cerebrovascular Diseases 2016;24(2):134-136
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.
9.Headache in children w ith moyamoya disease
Rui ZHANG ; Zhengshan ZHANG ; Lian DUAN
International Journal of Cerebrovascular Diseases 2016;24(2):128-130
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.
10.Moyamoya disease associated w ith intracranial aneurysm:clinical features, risk factors of hemorrhage and treatment outcomes
Huaitao YANG ; Zhengshan ZHANG ; Lian DUAN
International Journal of Cerebrovascular Diseases 2015;(2):102-106
ObjectiveToinvestigatetheclinicalfeatures,riskfactorsforbleedingandtreatment outcomes in moyamoya disease patients w ith intracranial aneurysms. Methods The clinical symptoms, location and size of aneurysm, treatment and the long-term folow-up results of the moyamoya disease patients w ith intracranial aneurysms w ere analyzed retrospectively. Results A total of 34 moyamoya disease patients w ith intracranial aneurysms (35 aneurysms) w ere enrol ed, including 22 (64.7%) in the intracranial hemorrhage group and 12 ( 35.3%) in the non-intracranial hemorrhage group. Of the 35 intracranial aneurysms, 23 (main artery type 11, peripheral artery type 12) w ere in the intracranial hemorrhage group and 12 (main artery type 11, peripheral artery type 1) w ere in the non-intracranial hemorrhage group. There w ere 29 smal aneurysms and 6 medium aneurysms (al w ere patients w ith hemorrhagic moyamoya disease). The aneurysms w ere mainly peripheral arterial type in the intracranial hemorrhage group, and the aneurysms w ere mainly artery type in the non-intracranial hemorrhage group. There w as significant difference in aneurysm typing betw een the tw o groups ( P= 0.013 ). Tw o patients did not perform encephalo-duro-arterio-synangiosis (EDAS) in the intracranial hemorrhage group, other patients and those of the non-intracranial hemorrhage group performed EDAS. Angiographical reexamination revealed that 3 patients w ith peripheral aneurysm disappeared, and 1 aneurysm recurred after aneurysm embolization, and the remaining aneurysms did not have any change. Long-term fol ow-up show ed that 1 patient died of sudden cerebral hemorrhage at 1 year after procedure in the intracranial hemorrhage group, and the others did not have ischemic or hemorrhagic stroke. The modified Rankin scale scores w ere improved in 21 patients. Conclusions There are differences in moyamoya disease patients w ith intracranial aneurysm typing w ith different clinical manifestations. Moyamoya disease patients w ith intracranial aneurysms are mostly smal aneurysms and they can not temporarily be treated directly and can perform EDAS directly. Intracranial aneurysms after procedure may remain long-term stability, and some peripheral aneurysms may disappear.