2.Establishment of a rapid method for synthesis and detection of gRNA
Jianyong DU ; Ran DENG ; Hong GAO ; Weidong YONG
Acta Laboratorium Animalis Scientia Sinica 2015;(3):267-271,290
Objective The purpose of this study was to establish a rapid method for synthesis and detection of guild RNA (gRNA) which is an essential component in CRISPR/Cas9 knockout technology .Methods First, the Nkp46 gRNA core fragment was synthesized as amplification template .Second , the forward and reversed primers of the matched gRNA were designed using the Nkp46 gene as reference sequence .Third, the DNA fragment of Nkp46 gRNA was amplified by PCR technology using the synthesized gRNA core fragment as template .The gRNA was reversely transcribed in vitro u-sing amplified DNA fragment as template .The efficiency and specificity of gRNA and its interaction with Cas 9 were detec-ted in vitro.Results The specificity and activity of Nkp46 gRNA were high .The obtained gRNA interacted with Cas 9 en-zyme and successfully cut the target double-stranded DNA at the designed site .Conclusions The method for synthesis and detection of gRNA established in this study is faster than the original method , and the created gRNA is fully functional .
3.Direct Three-dimensional Contrast-Enhanced MR Venography in Assessment of the Lower Extremity Veins
Hong DU ; Yong ZHANG ; Tianping ZHAO ; Wei ZHANG ; Leizhen YANG
Journal of Practical Radiology 2001;0(07):-
Objective To evaluate the three-dimensional contrast-enhanced MR venography in assessment of the lower extremityveins.Methods 15 patients with lower extremity veins thrombosis confirmed by operation,DSA or ultrasonic were undergone directthree-dimensional contrast-enhanced MR venography using GE Signa 1.0T MR system.The demonstration of the lower extremity veins wasobserved.Results 3D deep or superficial vein images of low abdomen,pelvis and low extremity were acquired satisfactorily in all 15patients.Conclusion Direct three-dimensional contrast-enhanced MR venography is a favorable method in accessing lower extremity veins.
4.Relationship between Fatty Liver and Blood Lipids in Children with Simple Obesity
shao-ling, WANG ; yong-hong, CHEN ; jun-bao, DU
Journal of Applied Clinical Pediatrics 2004;0(07):-
Objective To study the relationship between fatty liver and blood lipids in children with simple obesity and the precaution of this disease.Methods Forty-eight cases from 2 to 16 years old with simple obesity were divided into 2 groups.The first group included 19 cases with fatty liver(38.78%);the second group included 29 cases without fatty liver(61.22%). Blood lipids in 2 groups were analyzed with biochemical method.Blood total cholesterol(TC),triglyeride(TG),high density lipoprotein(HDL) and low density lipoprotein(LDL) were detected and the liver was examined with ultrasound.Results Blood TC,TG,HDL and LDL in cases with fatty liver were higher than those wit-hout fatty liver.Blood TG had the closest relation with fatty liver(P_a
5.Analysis of Clinical Symptoms and Blood Lipid Components in Simple Obesity Children
A-YI-GU-LI ; yong-hong, CHEN ; jun-bao, DU
Journal of Applied Clinical Pediatrics 2004;0(07):-
Objective To analyze blood lipid components in simple obesity children and to explore the effects of obesity in the lipid metabolism and its clinical significance.Methods A total of 90 children,including 50 simple obesity children(obesity group)and 40 normal children(control group),were enrolled in this study.The age ranged from 2.5 to 16.0 years.Their blood lipid profiles of all the children were analyzed.The blood lipid profiles were examined by biochemical analysis,including triglyeride(TG),total cholesterol(TC),low density lipoprotein(LDL) and high density lipoprotein(HDL),and the livers of all the children were analyzed.The blood lipid profiles were examined by ultrasonograph.Results 1.There were no significant differences in age and height in obesity group and control group,but there were significant differences in body mass index(BMI) and blood pressure(P0.05),which had no statistical meaning.3.Liver ultra sonogram showed that 18 cases had fatty liver(36%) in simple obesity children.Conclusions Metabolic disorder of blood lipid is present in simple obesity children,who have a tendency to get fatty liver.LDL is markedly elevated in obesity group.Arteriosclerotic cardiovascular disease should be prevented at earlier period of childhood.
6.Preliminary investigation of Suidasia nesbitti breeding status in ground dust flour in Qiqihaer City
Yong HONG ; Fengxia DU ; Shaosheng WANG ; Chaopin LI
Chinese Journal of Schistosomiasis Control 2017;29(2):225-227
Objective To investigate the Suidasia nesbitti breeding status in the ground dust flour collected in Qiqihaer City. Methods Totally,16 aliquots of ground dust flour were respectively collected from a college canteen and 15 households in Qiq-ihaer City. Then 10 g dust flour was taken from individual sample for isolation of the mites that were made of slide specimen,and the mites were identified and classified under a microscope. Results Acaroid mites were found in 15 of the 16 aliquots of sam-ples(detection rate being 93.8%),and 7 species,belonging to 6 genera under 3 families,were identified. A total of 561 heads of mites were isolated from 160 g samples,with an average breeding density of 3.51 heads/g. The most breeding mite was associ-ated with S. nesbitti. Conclusions Various species of mites are breeding in the ground dust flour in Qiqihaer City,and S. nesbit-ti occurs the most. These findings indicate that effective measures should be taken to prevent and control the harm associated with acaroid mite contamination.
7.Research and application of incorporating innovative research into undergraduate training
Yong-hong DU ; Jian-zhong ZOU ; Zhi-biao NG WA ; Jin BAI ; Jia-zhen YAN
Chinese Journal of Medical Education Research 2011;10(7):777-779
To introduce scientific research innovation into undergraduate education, cultivating innovative talents has been an urgent mission of current higher education. This article reviewed our experience, with the introduction of producing-studying-researching platform built on original innovative achievements of Chongqing medical university,of training physical medicine physician to be practical talents of large-scale diagnostic and therapeutic medical equipments, and was aimed to explore introducing producing-studying-researching platform into undergraduate education as well as improve personnel training quality of undergraduates.
8.Construction and identification of human tissue kallikrein gene eukaryotic expressing vector.
Yong, DAI ; Wujian, PENG ; Tiyuan, LI ; Hong, DU ; Wenxue, SUN ; Deheng, CHEN ; Zhuojia, XU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(2):164-6
To clone and sequence the human tissue kallikrein gene of Chinese, and to construct eukaryotic expression recombinant of KK, total RNA was extracted from human pancreas and human tissue kallikrein gene cDNA was amplified by PCR after reverse-transcription by using Oligo(dT) primer. The original kallikrein cDNA was recovered and filled with Klenow enzyme and inserted into KS plasmid. After restriction endonuclease digestion, KK cDNA was sequenced by ABI377 analyzer. Then the KK gene was amplified from pBluescript KSKK and inserted into pcDNA3. A sequence comparison showed that the cloned kallikrein gene was only one nucleotide different from that reported in the Genbank. The coding amino acid was Asp in the Genbank gene, while the coding amino acid of Chinese kallikrein gene was Asn. The KK cDNA fragment was inserted into the eukaryotic expression vector pcDNA3. The cloned kallikrein gene and the pcDNA3KK can be used for further study in gene therapy.. .
9.Analysis of the Etiology of Pulmonary Arterial Hypertension in 276 Children
xue-qin, LIU ; jun-bao, DU ; yong-hong, CHEN ; yu-wen, MA ; wan-zhen, LI
Journal of Applied Clinical Pediatrics 2006;0(13):-
Objective To investigate the etiology and clinical characteristics of pediatric pulmonary arterial hypertension(PAH) and improve its early diagnosis and treatment.Methods The clinical and echocardiogram data of all inpatients with PAH in Pediatric Department of Peking University First Hospital between May 1995 and May 2007 were retrospectively analyzed for age,sex,etiology,symptoms and echocardiographic measurement of pulmonary artery pressure.Data were divided into groups according to different etiology and statistics.Pulmonary arterial systolic pressure(sPAP) values estimated from the tricuspid regurgitant velocity by Doppler echocardiography were compared among different groups.Cases who were not belonged to the first category of the Venice Clinical Classification of pulmonary hypertension were not included.Results Totally 276 cases,168 boys and 108 girls were diagnosed to have PAH.Age ranged from 1 month to 17 years,median age was 9 months.Most of pediatric PAH was associated-PAH(267 cases,96.7%),while idiopathic PAH took a small part(9 cases,3.3%).Congenital heart disease-associated PAH(CHD-PAH) was predominant(245 cases,88.7%) and left to right shunt was the main lesion (217 cases,88.6%),while complex lesion-associated PAH comprised 28 cases(11.4%).Connective tissue disease associated PAH(CTD-PAH) was the second common among this group of pediatric PAH patients(19 cases,6.9 %).The incidence of PAH in systemic lupus erythematosus(SLE),juvenile rheumatoid arteritis and takayasu arteritis were 10.3 %(13/126),8.7%(4/46),15.4%(2/13),respectively.The other 3 cases of PAH were associated with portal hypertension(2 cases) and thalassanemia(1 case).The estimated sPAP from tricuspid regurgitant velocity in 8 cases with idiopathic PAH[(74.6?23.9) mmHg(1 mmHg=0.133 kPa)]was higher significantly compared with those of 33 cases of CHD-PAH [(58.0?19.7) mmHg ] and 12 cases of CTD-PAH [(49.6?18.9) mmHg] respectively(t=-2.052,-2.609 Pa
10.Diagnosis of Postural Orthostatic Tachycardia Syndrome in Children
A-YI-GU-LI ; jun-bao, DU ; qing-you, ZHANG ; yong-hong, CHEN
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To discuss the clinical diagnostic method of postural orthostatic tachycarda syndrome(POTS) in children.Methods Thirty-six children with POTS were selected for the research.Among them 15 were boys,21 cases were girls.The age ranged from 5.9 to 16 years,average age 12.3 years.Among them 28 patients(78%) were in between 11-16 years.The age distribution,clinical courses and hemodynamic indexes were analyzed and also the incidence of clinical manifestations and investigation reports were observed.Results Among 81 patients of orthostatic regulation disturbance,36 patients were diagnosed POTS,which was 47% of total.The clinical courses ranged from 1 day to 5 years,average clinical course 10.2 months.The clinical courses of more than half of the total patients were within 6 months(56%).The common clinical features of POTS were chest tightness on standing,vertigo,fatigue,palpitation,syncope,orthostatic regulation disturbance.Ten patients were also associated with gastrointestinal symptoms like nausea,vomiting.The most common feature of POTS patients was tachycardia(HR increased by ≥30 times/min) within 10 min after head-up tilt test(HUT).Average HR increased by 38 times/min.In some patients HR increased up to ≥120 times/min.There were no significant changes in blood.In 23 cases(64%),the T waves were descended by ≥0.2 mV in 2 or more than 2 leads in ECG reading.Investigations reports showed that there were 12 cases whose urine specific gravity was increased.In 11 cases HCO_3~-decreased.Conclusions POTS is commonly seen in schooling female children.The common symptoms are vertigo,chest tightness,fatigue,palpitation.HUT is an important method for the diagnosis of POTS.