2.Influence of 7-dehydrocholesterol reductase gene silencing on the fusion of mouse palatal shelves.
Wenlin XIAO ; Cuizhu ZHUANG ; Yan SHI ; Yaoxiang XU ; Lingfa XUE
West China Journal of Stomatology 2015;33(1):29-34
OBJECTIVERNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves.
METHODSThe pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV- siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n = 10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n = 10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experimental group (n = 10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses.
RESULTSSEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves. RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a significant difference (P < 0.05).
CONCLUSIONResults indicate that Dhcr7 gene silencing affects the fusion of palatal shelves. Thus, Dhcr7 gene may serve a function in the normal development of palates.
Animals ; Cleft Palate ; Gene Silencing ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Organ Culture Techniques ; Oxidoreductases Acting on CH-CH Group Donors ; Palate ; growth & development ; RNA, Messenger
3. Preparing an animal model of critical femoral defect in rabbit femoral condyle and the critical bone defect size
Chinese Journal of Tissue Engineering Research 2020;24(20):3191-3195
BACKGROUND: Rabbit model of distal femoral bone defect has been widely used to test bone tissue engineering materials for bone defects. However, there is no uniform standard for the size of the cylindrical bone defect model of the rabbit femoral condyle, which ranges 5-9 mm in diameter and 8-12 mm in depth. OBJECTIVE: To establish the bone defect model of adult rabbit femoral condyle with different sizes and to determine the critical bone defect size of the femoral condyle METHODS: Eighteen male New Zealand White rabbits aged 6 months were randomly divided into three groups according to the diameter of bone defect: 5 mm diameter group, 6 mm diameter group, and 7 mm diameter group. The defect depth was 10 mm. These rabbits underwent bilateral radial surgery, a total of 12 sides. Computed Tomography (CT) scan and three-dimensional reconstruction were performed at 1 day, 4, 8, 12 weeks after surgery. The CT-Hedberg score was used to evaluate the healing of bone defects. The rabbits were sacrificed at 12 weeks after surgery, and the femoral condyle specimens were taken out. Healing of the defect was analyzed by gross observation and hematoxylin-eosin staining. The study protocol was approved by the Animal Ethics Committee of Xuzhou Medical University. RESULTS AND CONCLUSION: All rabbits survived after surgery. The gross observation showed that the defect of 5 mm diameter group was filled with new bone tissue, the femoral condyle was well shaped, and the bone defect was completely repaired. In 6 mm and 7 mm diameter groups, depressed deformation was obviously observed in the defect area, with less new bone tissue, and the defect was was not repaired. The CT images showed that the defect area of 5 mm diameter group gradually decreased, and the broken ends of the defect were bridged. In the defect area of 6 mm and 7 mm diameter groups, only a small amount of new bone tissue was implanted, and the defect area was slightly reduced. At the 12th week after surgery, the cortical bone structure of 5 mm diameter group was intact and continuous, the femoral condyle was well shaped, and the bone defect was completely repaired. The defects of 6 mm and 7 mm diameter groups were partially or not repaired, and the defect cavity was still visible in the 7 mm diameter group. The CT-Hedberg scores of 6 mm diameter group were significantly lower than those of 5 mm diameter group at different time points (P < 0.05), and there was no significant difference in the CT-Hedberg scores between 6 mm and 7 mm diameter groups (P > 0.05). Histological results showed that there were irregular trabecular structures in the defect area of 5 mm diameter group, with a large amount of new bone tissue. In the other two groups, there were some new bone trabeculae around the bone defect, but the defect area was less filled with new bone tissue. During the 12-week observation period, the femoral condyle defect with a diameter of > 6 mm and a depth of 10 mm could not heal spontaneously, while the defect with a diameter of < 6 mm could be completely repaired, which met the criteria of critical bone defect. Therefore, the diameter of < 6 mm could be used as the critical bone defect size of rabbit femoral condyle.
4.MRI study of relationship between secondary spinal canal stenosis and subacute combined degeneration
Weipeng CHEN ; Haitao LIN ; Peixu YAO ; Ruyao ZHUANG ; Shi ZHOU ; Xiangting ZENG ; Qiuyu XU
Journal of Practical Radiology 2016;32(10):1506-1509
Objective To investigate MRI characteristics of subacute combined degeneration(SCD)with secondary spinal canal stenosis.Methods The clinical and MRI imaging data of 56 patients with SCD were collected to analyze the performance characteristics between spinal cord lesions and spinal canal stenosis,which depended on the synergism of lumbar disc bluge or herniation,degenerative thickening of the ligament flavum and posterior longitudinal ligament.Results Among 56 SCD cases underwent MRI scan,45 cases were combined with spinal cord lesions which showed typical signs of SCD.37 patients were secondary spinal canal stenosis with typical signs,but 2 showed no typical signs.8 patients were no secondary spinal canal stenosis and showed typical.9 cases showed neither spinal cord lesions nor secondary spinal canal stenosis.There was significant difference (P <0.05)between relative secondary spinal canal stenosis and spinal anomaly signal.The course of 1 5 cases were shortened after treated by physical in 37 cases of SCD with secondary spinal canal. Conclusion The secondary spinal canal stenosis can cause microcirculation dysfunction of the spinal cord,which is a key factor contributing to the imaging manifestation.
5.Prevalence and Related Factors of Anxiety and Depression in Persons with Disabilities in Shanghai
Zhuang TIAN ; Yan BAI ; Jumi XU ; Ruirui TANG ; Mei SUN ; Gang CHEN ; Shenxun SHI ; Jun Lü
Chinese Journal of Rehabilitation Theory and Practice 2016;22(3):326-329
Objective To investigate the prevalence of anxiety and depression among the persons with disabilities in Shanghai and the factors related to them. Methods From November to December, 2014, 731 persons with disabilities were consecutively enrolled in this cross-sectional study. They were assessed with Self-rating Anxiety Scale (SAS) and the Self-rating Depression Scale (SDS). Binary Logistic regression analysis was employed to reveal potential related factors of anxiety and depression. Results The prevalence rates of anxiety and depression among the disabled in Shanghai were 9.58%and 8.48%, respectively. The duration (OR=0.967, P<0.05) and severity (OR<1, P<0.05) of disability were associated with depression, whereas retiral (OR=2.047, P<0.05), living alone or in care unit (OR=3.073, P<0.01) and duration of disability (OR=0.956, P<0.01) were associated with anxiety. Conclusion The frequency of anxiety and depression is mild among the persons with disabilities in Shanghai, and a special intervention is needed.
6.Vitamin D status in healthy adults in Guiyang of China
Qiao ZHANG ; Nianchun PENG ; Shujing XU ; Miao ZHANG ; Huijun ZHUANG ; Jinglu ZHANG ; Liu LIU ; Lixin SHI
Chinese Journal of Endocrinology and Metabolism 2011;27(8):659-663
Objective To evaluate vitamin D status in healthy adults living in Guiyang. Method 1 500 healthy volunteers aged 20-79 years ( mean 45.2 years ) were recruited from a community in Guiyang by cluster sampling method. Questionnaires for living habits and fasting blood samples were collected from November, 2009 to February, 2010. Serum 25 ( OH ) D concentrations were measured by radioimmunoassay, using the DiaSorin kit,USA. Results The average serum 25 ( OH ) D level was ( 20. 4±9.0 ) ng/ml. The percentages of vitamin D deficiency [25 ( OH ) D < 20 ng/ml], insufficiency [20 ng/ml ≤ 25 ( OH ) D < 30 ng/ml], and sufficiency [25 ( OH ) D ≥ 30ng/ml] were 52. 3% , 32. 3% , and 15.4% , respectively. The 25 (OH) D concentrations in the young, middle-aged,and elderly were ( 18. 2±9. 2), (22. 8±8. 7), and ( 19. 9±7.8) ng/ml, respectively. The percentages of vitamin D deficiency in these groups were 62. 8%, 40. 2%, and 55.4%, being 61.6% in higher educational group ( ≥ 13 years) and 64. 4% in the group with lower body mass index ( < 18.5 kg/m2 ). Conclusion Vitamin D deficiency is common in Guiyang including all age groups, especially among the youth and the elderly. Serum 25-hydroxyvitamin D level is also influenced by education, age, smoking, and other factors.
7.Association of single nucleotide polymorphisms in inflammation-related genes with amnestic mild cognitive impairment
Liying ZHUANG ; Zhijun ZHANG ; Chunxian YUE ; Hao SHU ; Xiaoyan LIU ; Xiaohui XU ; Hui YU ; Yongmei SHI
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(5):410-413
ObjectiveTo examine whether the single nucleotide polymorphisms in inflammation-related genes are associated with the risk of amnestic mild cognitive impairment (aMCI).MethodsThe study recruited 116 aMCI patients and 93 matched healthy controls.All subjects underwent extensive assessment of cognitive function,genotyping was carried out on the platform of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.Results ( 1 ) There was prominent discrepancy between aMCI and controls in the memory,attention and executive functions,20 minutes delayed recall of auditory verbal memory test (AVMT) (3.0(0.0 ~ 10.0 ),8.0 (0.0 ~ 12.0),t =- 8.533,P < 0.05 ),recall of Rey-Osterrieth complex figure test ( R-O CFT) (11.2 ±8.3,16.1 ±8.0,t=4.216,P<0.05),digit span test (DST) (12.0(7.0 ~ 19.0),13.0(7.0 ~20.0),Z=-2.516,P<0.05),trail making test A (TMTA) (80.0s(35.0 ~200.0)s,72.0s(29.0 ~512.0)s,Z=-3.113,P<0.05),trail making test B (TMTB) ((180.1 ±72.7)s,(141,7 ±52.1)s,t=-4.385,P<0.05 ).(2) No significant differences were found in frequencies of alleles,genotypes and hapolotypes of inflammation mediator genes ( interleukin 10,interleukin 1 A,interleukin 1 B,tumor necrosis factor,interleukin 6,α1- an-tichymotrypsin gene,transforming growth factor B1 ) between aMCI and controls (P > 0.05 ).ConclusionThe results indicate that polymorphisms in the inflammation-related candidate genes do not appear to be involved in the risk of developing aMCI.
8.Percutaneous kyphoplasty for treatment of osteoporotic vertebral fractures and collapse with intravertebral vacuum sign
Qingshan ZHUANG ; Zhaowan XU ; Qinmin WU ; Feng SHI ; Xubin JI ; Guoxia SUI
Chinese Journal of Trauma 2013;29(7):608-613
Objective To investigate the clinical effect of percutaneous kyphoplasty (PKP) in treatment of osteoporotic vertebral fractures and collapse with intravertebral vacuum sign.Methods A retrospective study was conducted on the clinical and radiological data of 31 patients with osteoporotic vertebral fractures and collapse with intravertebral vacuum sign treated by PKP from June 2009 to June 2011.Vertebrae body variation,visual analog scale (VAS) and Oswestry disability index (ODI) before operation,during follow-up at postoperative one week and at postoperative 3-6 months were used as outcome measurements.Results All the patients went through operations successfully and presented significant mitigation of low back pain in 24 hours after operation.The vertebrae body height at postoperative one week [(17.2 ±4.2) mm] and at postoperative 3-6 months [(16.8 ±5.1)mm] were statistically different from that before operation [(11.4 ± 1.7) mm,P <0.01],while there was no statistical difference between the two follow-ups (P > 0.05).VAS and ODI at postoperative one week [(2.8 ± 1.7) points and (31.6 ± 8.4) points] were statistically different from those before operation [(8.6 ± 1.3) points and (78.3 ±8.5) points,P<0.01].VAS and ODI at postoperative 3-6 months [(2.3 ±0.8) points and (23.7 ± 2.3) points] presented statistical differences from those before operation (P < 0.01),but no statistical differences from those at postoperative one week (P > 0.05).Conclusion PKP obtains satisfactory clinical outcomes,for it relieves low back pain and restores vertebral body height.
9.Association study of candidate gene polymorphisms of tau protein pathway with amnestic mild cognitive impairment
Xiaoyan LIU ; Zhijun ZHANG ; Yongmei SHI ; Hui YU ; Chunxian YUE ; Hao SHU ; Liying ZHUANG ; Xiaohui XU
Chinese Journal of Neurology 2012;45(2):78-83
Objective To investigate the relationship between amnestic mild cognitive impairment and functional genes associated with hyperphosphorylated tau protein.Methods One hundred and sixteen amnestic mild cognitive impairment (aMCI) patients and 93 normal controls were recruited for the study.Multi-dimension neuropsychologic tests were used to assess the cognitive function extensively.MassARRAY and iPlex systems were used to measure candidate SNP polymorphisms,analyze genotypic,allelic or haplotypic distributions and their interaction with ApoE ε4 and the correlation with the cognitive function in the subjects.Results ( 1 ) The scores of neuropsychologic tests in memory domain ( Auditory Verbal Learning Test (AVLT)-first immediate recall,AVLT-second immediate recall,AVLT-second immediate recall,AVLT-5 minute delayed recall,AVLT-20 minute delayed recall,AVLT-recognition,Rey-Osterrich Comolex Test-delay) in aMCI patients ( 3.0 ( 0-7.0 ),5.0 ( 1.0-10.0),6.0 ( 1.0-11.0 ),4.0 (0-11.0),3.0(0-10.0),20.0(8.0-24.0),11.2 ±8.3) were significantly lower than those in the normal controls(4.0(0-9.0),7.0(2.0-11.0),9.0(3.0-12.0),8.0(0-12.0),8.0(0-12.0),22.0 (10.0-24.0),16.1±8.0) (Z=-3.592,-6.802,-6.408,-8.173,-8.533,-5.647 andt=4.216 respectively,all P <0.01 ) ; (2) Genotypic distributions of rs242562 GG in aMCI (7.826% ) were significantly lower than those in normal controls (20.65%,OR =0.3525,95% CI 0.1411-0.8807,P =0.024 98),however there were no differences in the genotypic,allelic or haplotypic distributions between aMCI patients and controls of glycogen synthase kinase-3β,cyclin dependent protein kinase-5,calcium and calmodulin-dependent protein kinase-Ⅱ,cell division cycle 2,dual-specificity tyrosine-phosphorylation regulated kinase 1A and low density lipoprotein receptor-related protein 6; (3) MAPT/STH rs242562 genotype was correlated with AVLT-immediate recall,AVLT-delayed recall,Rey-Osterrieth Complex Test,Rey-Osterrieth Complex Test-delayed recall and Clock Drawing Test (H =9.763,12.258,10.508,9.624,10.767,F =3.700,3.123 and H =6.591 respectively,all P < 0.05 ) ; (4) There were no differences in the distributions of MAPT/STH rs242562 GG genotype and ApoE ε4 haplotype between aMCI patients and normal controls.Conclusions MAPT/STH rs242562 GG genotype decreases the genetic risk of aMCI,which might have important role in memory function in aMCI.The interaction between rs242562 GG and ApoE ε4 doesn' t affect the susceptibility to aMCI.
10.Characteristic of peripheral blood monocyte subsets and chemokines in early stage of acute coronary syndrome
Miaomiao XU ; Wei SHEN ; Haiming SHI ; Xinyu ZHUANG ; Xiangxu LIU ; Yang OU ; Shengjia SUN ; Bangwei WU ; Zhidong ZHU ; Yufei CHEN
Fudan University Journal of Medical Sciences 2017;44(4):403-409,416
Objective To investigate the expression of monocyte subsets and their chemokine,i.e.,monocyte chemoattractant protein (MCP-1) and fractalkine (FKN),in patients with acute coronary svndrome (ACS),and to analyze their correlation.Methods Patients with the syndrome of pectoralgia and to be inspected with coronary angiography (CAG) in our hospital from Sep.to Dec.,2016 were included.Patients' venous blood was collected on the operation day before operation,the level and proportion of monocyte (Mon) subsets,which was namely CD14 + CD16-Mon (Mon1),CD14+CD16 + Mon (Mon2) and CD14-CD16 + Mon (Mon3) according to the expression of cluster differentiation-14 (CD14) and CD16,were detected by flow cytometry (FCM).Patients' venous blood was collected on the operation day before operation and one day after operation,the concentrations of MCP-1 and FKN in plasma were measured by ELISA.We compared the expression levels of MCP-1-Mon1 and FKN-Mon3,and analyzed their relationship between each other respectively in different groups.Results Diagnosed according to the clinical symptoms,myocardial markers,electrocardiogram and CAG results,70 individuals were analyzed,including 30 patients with acute myocardial infarction (AMI group),25 patients with unstable angina pectoris (UAP group) and 15 patients with the chest pain symptoms and normal CAG results (control group).The percentage of Mon1 in the AMI group was higher than that in the other groups (P<0.05);no difference was observed for Mon3 among the groups (P>0.05).The Mon3/Mon1 ratio in the AMI group was lower than that in the control group (P<0.05).Moreover,the levels of FKN and MCP-1 in the ACS group were greater than those in the control group.The level of red blood cell distribution width (RDW) was significantly increased in the AMI and UAP group than that in the control group (P<0.05).There was a significant correlation between FKN and Mon3 (P<0.05,R=0.650 2).Conclusions The monocyte subset of Mon1 and Mon3 increased in the early stage of ACS,with their chemokine (FKN and MCP-1) increasing at the same time.There is a significant correlation between FKN and Mon3,which indicates MCP-1-Mon1 and FKN-Mon3 may participate in the pathophysiological process of early ACS in patients.