1.Comparison between the quantitative and semi-quantitative fluorescence RT-PCR assays for the detection of XDH/XO mRNA expression in different organ tissues of rhesus monkey
Donghong TANG ; Yousong YE ; Zheli LI ; Bo PENG ; Guizhen LI ; Runping LI ; Guangrui YANG
Chinese Journal of Comparative Medicine 2015;(12):47-53
Objective To analyze the differences between the semi-quantitative RT-PCR and real time quantitative fluorescence RT-PCR assays for detecting XDH/XO mRNA expression in various organ tissues of rhesus monkey, and provide useful reference in methodology of experimental studies.Total RNA was extracted from the myocardium, kidney, testis, skin, and liver tissues, respectively, for detecting XDH/XO mRNA expression in rhesus monkey by semi-quantitative RT-PCR and real time quantitative fluorescence RT-PCR assays.The sensitivity and specificity of the two assays were compared with each other using the same primer sequences and reference genes.Results The expression of XDH/XO mRNA in different organ tissues were detected by both the two PCR assays.The sensitivity of quantitative fluorescence real-time RT-PCR for the XDH/XO mRNA expression in the liver tissue was 39 times higher than that by semi-quantitative RT-PCR.Conclusions Both the quantitative and semi-quantitative fluorescence RT-PCR assays can be used to detect the expression of XDH/XO mRNA in different organ tissues of rhesus monkey.The sensitivity of quantitative fluorescence real-time PCR assay is more sensitive than that of the semi-quantitative RT-PCR assay.
2.Association between miR146a(rs2910164)G>C polymorphism and susceptibility to acute lymphoblastic leuke-mia in children
Xiaolei LIU ; Liying LIU ; Zheli CAO ; Baozhen GUO ; Mingwei LI
Chinese Journal of Applied Clinical Pediatrics 2018;33(3):200-202
Objective To investigate the association between miR146a(rs2910164)G>C polymorphism and susceptibility to acute lymphoblastic leukemia(ALL)in children.Methods Two hundred blood specimens were ob-tained from children with ALL as patient group and 100 blood specimens were obtained from healthy children as healthy control group,who were all from Baoding First Central Hospital between March 2010 and October 2016.There were no significant differences in sex and age between patient group and healthy control group(χ2=0.430,P=0.512;χ2=2.839,P=0.092).The distribution of gene frequency of patient group and healthy control group conformed to Hardy-Weinberg equilibrium.miR146a(rs2910164)G>C polymorphism was identified by adopting restriction fragment length polymorphism(RFLP).The relation of genotype and ALL was demonstrated by odds ratio(OR)and 95% credibility interval(CI). Results Gene frequency of miR146a(rs2910164)GG,GC and CC genotypes in patient group and healthy control group was 16.0%,44.5%,39.5% and 29.0%,41.0%,30.0%,respectively.The GC/CC genotypes were significantly higher in patient group than those in healthy control group(GG genotype as reference,GC genotype:OR=1.967,95%CI:1.054-3.672,P=0.037;CC genotype:OR=2.386,95%CI:1.239-4.595,P =0.012). Conclusion miR146a(rs2910164)G>C polymorphism is significantly associated with susceptibility to ALL in chil-dren.
3.Exploration of establishing a tree shrew model of chronic gastrointestinal mucosal injury
Yousong YE ; Jiahong GAO ; Ru ZHAO ; Zheli LI ; Chenyun WANG ; Kaili MA ; Donghong TANG
Chinese Journal of Comparative Medicine 2018;28(3):43-47
Objective To explore the feasibility of establishing a tree shrew model of chronic gastrointestinal mucosal injury. Methods A total of 12 adult male tree shrews were randomly divided into 3 groups. The experimental groups 1 and 2 were administered with intraperitoneal injection of 2 mg/(kg·d)and 1 mg/(kg·d)of 1-methyl-4-phenyl-1,2, 3,6-tetrahydropyridine(MPTP)once every day for 56 days, respectively. The control group was given the same volume of sterile saline at the corresponding time points. Changes in the body weight of the tree shrews were observed. The contents of dopamine in the cerebrospinal fluid were detected. Gastrointestinal morphology was observed by stereoscope and histopathological changes of the gastrointestinal mucosa were examined by HE staining. Results The body weight and the contents of dopamine in the cerebrospinal fluid of the tree shrews in the model group were significantly decreased(P< 0.05 for both). Pathological changes to some extent of the gastric antrum, the gastric body and the duodenum were observed, without obvious differences between the 2 mg/kg group and the 1 mg/kg group. No obvious changes were found in the control group. Conclusions Long-term intraperitoneal injection with a low dose of MPTP is a feasible method for the establishment of a tree shrew model of chronic gastrointestinal mucosal injury. The optimal dose is 2 mg/(kg·d)every day for 56 days.
4.The significance of urinary NGAL and KIM-1 in predicting the kidney injury in children with congenital heart disease
Jing WANG ; Xiuhong HU ; Hongjuan YANG ; Hongrui CUI ; Zheli NIU ; Mingming LI
International Journal of Laboratory Medicine 2017;38(19):2690-2693
Objective To investigate the significance of urine neutrophil gelatinase associated lipocalin (NGAL ) and renal injury molecule-1 (KIM-1) in predicting the acute kidney injury (AKI) in children with congenital heart disease after operation .Methods From April 2014 to December 2015 ,67 cases of cardiopulmonary bypass in children with congenital heart disease were studied in our hospital ,all patients were divided into AKI group (n=24) and non AKI group (n=43) by pRIFLE standard .Serum creatinine , urine NGAL and urine KIM-1 levels were compared between the two groups before and after the operation ,the receiver operating characteristic curve (ROC curve) and the area under the curve (AUC) were used to evaluate the value of NGAL and KIM-1 in pre-dicting the postoperative AKI in children with congenital heart disease .Results There was no significant difference between the two groups in preoperative and postoperative 2 h and 4 h creatinine (P>0 .05) ,but the levels of postoperative 12 ,24 ,48 h creati-nine in the non AKI group were significantly lower than those in the AKI group (P<0 .05) .The NGAL level of postoperative 2 ,4 , 6 ,12 h in non AKI group was significantly lower than that in AKI group (P<0 .05) ,but there was no significant difference in the level of postoperative 24 h urine NGAL between the two groups (P>0 .05) .There was no significant difference between the two groups of patients with postoperative 2 h urinary KIM-1 (P>0 .05) ,postoperative 4 ,6 ,12 ,24 h urinary KIM-1 levels in the non AKI group were significantly lower than those in the AKI group (P<0 .05) .The optimal time point separate detection of urinary NGAL levels to assist in diagnosis of AKI after 12 h ,AUC was 0 .834 (95% CI:0 .631-0 .912);the best time point separately to detect the level of KIM-1 AKI to assist in the diagnosis of AKI after 24 h ,AUC was 0 .871 (95% CI:0 .665-0 .933);combined de-tection of urinary NGAL and KIM-1 levels to assist the best time for the diagnosis of AKI after 24 h ,AUC was 0 .913(95% CI:0 .745-0 .968) .Conclusion Urine NGAL and urine KIM-1 in children with congenital heart disease after operation have important clinical significance in predicting the occurrence of AKI .
5.Biopsy of liver and kidney tissues in rhesus monkeys under B-mode ultrasound guidance
Zheli LI ; Yousong YE ; Shu ZHANG ; Lei FENG ; Chenyun WANG ; Qian CHEN ; Donghong TANG
Chinese Journal of Comparative Medicine 2018;28(3):78-83
Objective To establish a safe and effective method of rhesus monkey biopsy to take liver and kidney samples under B-mode ultrasound guidance. Methods A total of 4 adult monkeys(weight:8-12 kg; sex: male; age:11 -12 years old)were anaesthetized with 5 -10 mg/kg of ketamine hydrochloride for each through intramuscular injection. After successful anesthesia, abdominal shaving and iodophor disinfection, they were monitored from intercostal area of right upper quadrant or lateral waist subcostal abdomen portions to find liver or kidney organ by MyLab 30CV B-mode ultrasonography with 3.5 Hz transducer which was fixed with a guiding frame. Large vessels such as the portal vein and inferior vena cava were carefully avoided. The range of the biopsy gun was set to 15 mm. When the puncture target and the puncture needle were positioned in the guide line, the puncture target was perpendicular to the puncture needle, and then the trigger button of the puncture needle was pressed to obtain the liver or kidney tissue samples respectively. After puncture,the needle was pulled out quickly. The obtained liver and kidney tissues were used to extract RNA. Results About 13 mg of liver or kidney tissue was obtained by each puncture with volume convertion. This method was fast,reliable and safe,and the total RNA had high purity and integrity. There was no postoperative bleeding and infection. Conclusions This is a very important method for obtaining liver and kidney tissue samples of rhesus monkeys with the guidance of ultrasound. With this method, the research cost can be reduced, the life quality and animal welfare of laboratory non-human primates can be improved,and the accuracy of experimental result can be ensured.
6.Determination and comparison of the bone mineral density of different skeletal regions of female Wistar rats at different ages
Yousong YE ; Chenyun WANG ; Zheli LI ; Kaili MA ; Donghong TANG
Chinese Journal of Comparative Medicine 2018;28(4):65-68
Objective To provide reference values for bone mineral density(BMD)in different skeletal regions of female Wistar rats at different ages. Methods Thirty SPF female Wistar rats were selected. The BMD of different skeletal regions(skull,upper limbs,thighs,trunk,ribs,pelvis,spine and the whole body)was measured by dual energy X-ray absorptiometry(DXA)at 6,10,12,24 and 30 months of age. The bone mineral densities between different age groups and that of different skeletal regions in the same age groups were compared. Results The BMD of the skull,upper limbs,thighs,trunk,ribs,pelvis, spine and the whole body was increased rapidly with age, and reached a peak at 10 months of age. The BMD of the skull,upper limbs,thighs,trunk,ribs were significantly higher than the whole body BMD in the same month-age group(P< 0.05 or P< 0.01). However,there was no significant difference between the pelvic, spine and the whole body BMD(P> 0.05). There was a significant positive correlation among the three correlations(P<0.01). Conclusions Some background data are provided for the bone biology studies of female Wistar rats, and provide useful supplementary reference for the studies of bone metabolism in rats and their application in biomedicine.
7.The study and analysis of the effectiveness of "health education mobile classroom" in the outpatient department of children's hospital
Jingmin SUN ; Qunfeng LU ; Jiangjiang XU ; Hongye TIAN ; Jingjing CAO ; Hua DU ; Lijun CHEN ; Zheli ZHANG ; Youwei LI
Chinese Journal of Practical Nursing 2018;34(28):2222-2226
Objective To evaluate the effect of "health education mobile classroom " in the outpatient department of children's hospital, and to analyze the needs of the outpatient and family members on health education. Methods Through the retrospective analysis of the information of the outpatient and emergency patients in the children's specialist hospital to determine the content, the venue and the time period of the "health education mobile classroom". Selection and training the health educator to implement the health education. At the same time, push the training content through the mobile APP to the patient's family, to expand the education audience. To compare and analyze the patient satisfaction of the hospital and the satisfaction with the dimensions of the project. In the evaluation of the effect of health education and the influencing factors, the relationship between income, occupation and children is positively related, and age is negatively related to it. Results After the implement of this project, the satisfaction of the patient had increased for 3.65%(P=0.001). Overall assessment of the audience was very satisfied with (3.94 ± 0.25) points. In each dimension analysis, the satisfaction of the "training method"was the highest, at (3.94 ± 0.26) points. Satisfied with the attitude of the trainers and comprehensive ability were more than 3.80 points. Conclusions The "health education mobile classroom" can improve patient satisfaction in the outpatient department and emergency of children's specialist hospital. Audience has a certain degree of acceptance and satisfaction with this health education mode.