2.CAD model design for three-dimensional printing of tissue-engineered tooth scaffold
Jiayu ZHANG ; Xue MI ; Yi LIU ; Huiyu HE
Chinese Journal of Tissue Engineering Research 2015;(38):6195-6199
BACKGROUND:There are less reports on how to successfuly build the internal spatial configuration of tissue-engineered tooth scaffolds.
OBJECTIVE:To find a way to establish a series of three-dimensional digital modes for tissue-engineered tooth scaffold, such as CAD spatial configuration and Standard Template Library (STL) files.
METHODS:In order to get three-dimensional printing format of STL files, MICRO CT data of DICOM format were input into MIMICS and GEOMAGIC softwares, creating the outline of STL files. Then CATIA V5R17 software was used to create the three-dimensional digital mode of tissue-engineered tooth. Then, the overal model of the internal scaffold was obtained by arraying at the proper coordinates. Various overal scaffold configurations could be built rapidly by varying monomer configuration. The STL files of CAD model of three-dimensional printing tissue-engineered tooth were obtained by assembling the tooth outline mode and the internal scaffold.
RESULTS AND CONCLUSION: The CAD model was constructed successfuly, and this model could be directly used for three-dimensional printing rapid prototyping system to produce tissue-engineered tooth scaffolds. These findings indicate that the three-dimensional digital mode based on reverse engineering and positive engineering can be established, which can be used to quickly build a variety of internal spatial configurations of scaffold materials required for tissue-engineered teeth.
3.Recombinant lentiviral vector transfected sheep bone marrow mesenchymal stem cells and osteogenic gene expression changes
Xiangzhen HAN ; Huiyu HE ; Yang HU ; Jiaojiao BA ; Huanhuan WANG ; Xue MI
Chinese Journal of Tissue Engineering Research 2014;(6):821-828
METHODS:Lentiviral vectors carrying bFGF and BMP-2 were constructed to transfect sheep bone marrow mesenchymal stem cells. cells were divided into four groups:bFGF group, BMP-2 group, co-transfection group BACKGROUND:Basic fibroblast growth factor (bFGF) can promote the proliferation of bone marrow stromal cells, and bone morphogenetic protein-2 (BMP-2) has an important significance in the induction of new bone formation.
OBJECTIVE:To analyze the effects of bFGF, BMP-2 and their co-transfection on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and to compare the relative expressions of col agen I, osteocalcin and osteopontin before and after celltransfection, thereby providing theoretical implications for seed cells in the construction of tissue-engineered bone.
and control group. The RNA was extracted using real-time quantitative PCR to detect mRNA levels of col agen I, osteocalcin, and osteopontin.
RESULTS AND CONCLUSION:Significant difference in non-specific osteogenic gene expressions was found among the four groups (P<0.05). bFGF and BMP-2 showed an interaction (P<0.05). Expressions of col agen I and osteocalcin in the co-tranfection group were higher than those in the other three groups (P<0.05), but osteopontin expression exhibited no difference (P>0.05). In vitro experiments showed that the relative expression of col agen I, osteocalcin and osteopontin were higher in the co-transfection group, indicating the cells from the co-transfection group have strongest osteogenic capacity that are suitable for seed cells for bone tissue engineering.
4.Effect Of mechanical ventilation with the least delay on the severe cases of hand-foot-mouth disease
Qingling TIAN ; Baodong PANG ; Shuang ZHANG ; Huiyu ZHANG ; Xue FENG ; Yan LI ; Hongsheng GAO
Chinese Pediatric Emergency Medicine 2010;17(3):220-222
objective To explore the indication of mechanical ventilation and evaluate the effects of mechanical ventilation with the least delay on the Severe cases of hand-foot-mouth disease(HFMD).Methods Retrospective cohort study was conducted among the severe HFMD cases(n=66)admitted to pediatric intensive care unit(PICU)between July 2008 and september 2009.Sixty-six cases were divided into two groups:42 cases(group A,July 5 to October 31,2008)were ventilated with the common mechanical ventilation,24 cases(group B,April 21 to september 30,2009)were ventilated at least delay when the symptoms of neurogenic pulmonary edema appeared.Then the diffcrences on the survival rate and the mortality in the group A and group B were investigated.Results Twenty out of 42 patients died(47.6%),eighteen were cured(42.6%) and four showed improved signs(9.5%)in group A.Twenty-two out of 24 patients were cured(91.7%),two showed improved signs(8.3%)and no death in group B.The clinical effect of group B was much better tllan group A(P<0.001).Conclusion Early mechanical ventilation would improve the survival rate and decrease the mortality of severe hand-foot-mouth disease.
5.Study of the bone mineral density detected by dual-energy X-ray absorptiometry in 221 adolescents of Tianjin
Xiaolei SUN ; Xinlong MA ; Huiyu XUE ; Qian YANG ; Xueqing LIU ; Jing LU
Tianjin Medical Journal 2017;45(7):715-718
Objective To study the relationship between lumbar and proximal femoral bone mineral density (BMD) with gender,age and body mass index (BMI) in adolescents ranged from 12 to 19 years of age in Tianjin city,and provide preliminary reference to understand the bone health status of adolescent in the city.Methods The BMD values of lumbar spine (L1-L4,),femoral neck and total hip were detected by the dual-energy X-ray absorptiometry (DXA) in 221 adolescents from Tianjin Hospital,and the Z value was used for clinical diagnosis standard.Simultaneously the height and weight of adolescents were measured,and BMI was calculated.The differences of BMD values in the lumbar,femoral neck and total hip were compared between different age,gender and BMI groups.Results The BMD values of femoral neck and total hip were significantly higher in males than those in females (P ＜ 0.01).The BMD values of lumbar spine,femoral neck and total hip were gradually increased with age.The BMD was correspondingly increased in adolescents with higher BMI in the lumbar spine,femoral neck and total hip.Conclusion There is a close relationship between BMD with age,gender and BMI in adolescents.We should pay more attention to the nutrition health and the level of BMI in adolescents.
6. Effects and related mechanism of angiotensin-(1-7) on Toll-like receptor 4-mediated oxidative stress in human umbilical vein endothelial cells
Wenfeng YAN ; Jingjing XUE ; Huiyu YANG ; Bin LIANG ; Zhiming YANG
Chinese Journal of Cardiology 2017;45(3):223-229
Objective:
To explore the role and related mechanisms of angiotensin-(1-7)(Ang-(1-7)) on Toll-like receptor 4 (TLR4) mediated oxidized low-density lipoprotein(ox-LDL)-induced oxidative stress in human umbilical vein endothelial cells (HUVECs).
Methods:
HUVECs were cultured in vitro and divided into six groups: the control group (normal medium), the ox-LDL group(treated with 75 mg/L ox-LDL), the ox-LDL+ Ang-(1-7) group (1 μmol/L Ang-(1-7) pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL), the ox-LDL+ Ang-(1-7)+ A-779 group(1 μmol/L A-779 (Mas receptor) pretreated for 30 minutes, 1 μmol/L Ang-(1-7) pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL), the ox-LDL+ A-779 group (1 μmol/L A-779 pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL), the ox-LDL+ HTA125 group (10 μg/L HTA125 (TLR4-blocking antibody) pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL ). The corresponding index was detected after 24 hours after intervention. Apoptosis of cells were detected by Annexin V-FITC/PI double staining flow cytometry and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL). The generation of reactive oxygen species (ROS), products in oxidative stress, were detected by DCFH-DA staining. The mRNA and protein expression levels of NADPH oxidase 4(NOX4) and TLR4 were detected by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis respectively.
Results:
(1) The results of Annexin V-FITC/PI double staining flow cytometry showed that the proportion of apoptotic cells was higher in ox-LDL group than in control group ((21.18±1.40)% vs. (1.59±0.26)%,
7.Preparation of PTD4-Cu,Zn-SOD fusion protein
Shajie DANG ; Wenbo WEI ; Lichun HAN ; Wenbin ZENG ; Huiyu YUE ; Rongliang XUE
Chinese Journal of Anesthesiology 2017;37(8):939-942
Objective To prepare PTD4-Cu,Zn-SOD fusion protein.Methods The recombinant plasmid of pET 1 6b-Cu,Zn-SOD and pET16b-PTD4-Cu,Zn-SOD was transformed into Escherichia coli BL21 (DE3).Isopropyl β-D-1-thiogalactopyranoside was then added at a final concentration of 0.84 mmol/L,and the cells were incubated for 4 h to induce the expression of Cu,Zn-SOD and PTD4-Cu,Zn-SOD fusion protein.Lysozyme and ultrasound were used to lyse the bacteria,the supernatant was collected for 15％ SDS-PAGE to analyze the expression of the target protein.Ni-NTA His bind resin was used to purify Cu,Zn-SOD protein and PTD4-Cu,Zn-SOD fusion protein under natural conditions.Western blot was used to identify the target protein.Results The results of Western blot showed that the purity of the target protein was about 90％,and the Cu,Zn-SOD protein with a molecular weight about 19 kDa and the PTD4-Cu,Zn-SOD fusion protein with a molecular weight about 20 kDa were found.Conclusion PTD4-Cu,Zn-SOD fusion protein is prepared successfully.
8.Effects of Hcy and lncRNA ANRIL on atherosclerotic inflammatory response
Fen GAO ; Jie ZHU ; Hong LI ; Wen WEN ; Lijun CHEN ; Yaona LI ; Shuanqin XUE ; Huiyu YANG
Journal of Chinese Physician 2023;25(6):828-833
Objective:To study the relationship between the level of serum homocysteine (Hcy) and the antisense non coding gene (ANRIL) of long chain non coding RNA (lncRNA) cell cycle dependent kinase inhibitor 2B gene, and the effect on Atherosclerosis inflammation, that is, the expression of interleukin-10 (IL-10) and Monocyte chemoattractant protein-1 (MCP-1) in Human umbilical vein endothelial cell (HUVEC).Methods:HUVEC was cultured in vitro and cells were treated with different concentration gradients (blank control group, 0.5, 1.0, 2.0, 5.0 mmol/L) of Hcy. The expression level of lncRNA ANRIL was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of MCP-1 and IL-10. LipoFilter transfection reagents were used to transfect shANRIL and shNC into different cells, respectively. In the above experiment, the optimal Hcy concentration (5.0 mmol/L) was selected for intervention for 24 hours. Western blot was used to detect the protein expression levels of MCP-1 and IL-10.Results:After 24 hours of intervention with different concentrations of Hcy in HUVEC, Hcy significantly damaged endothelial cells, and the higher the Hcy concentration, the more severe the cell damage. Compared with the blank control group, the Hcy intervention group showed an increase in lncRNA ANRIL and MCP-1, while IL-10 decreased (all P<0.05); As the concentration of Hcy intervention increases, IL-10 decreases, while lncRNA ANRIL and MCP-1 increased (all P<0.05). Compared with the blank control group, the Hcy group, the shNC+ Hcy group, and the shANRIL+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). Compared with the shANRIL+ Hcy group, the Hcy group and the shNC+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). There was no statistically significant difference in the expression levels of IL-10 protein and MCP-1 protein between the shNC+ Hcy group and the Hcy group (all P>0.05). Conclusions:Hcy upregulates MCP-1 expression and downregulates IL-10 expression by promoting lncRNA ANRIL expression. Thus, it can promote cellular inflammatory reaction and participate in Atherosclerosis.
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