1.Isolation, culture and odontogenic phenotypes of human exfoliated deciduous dental pulp stromal cells
Zhengbin YAN ; Jingqiu HOU ; Tianxiang WANG
Chinese Journal of Tissue Engineering Research 2009;13(27):5305-5308
BACKGROUND: In the tooth regeneration and dental tissue engineering study, finding suitable stem cells are the central issues.OBJECTIVE: To isolate and culture child exfoliated deciduous tooth pulp stromal cells, and compare difference in dentin sialophosphoproteln expression prior to and following mineralization.DESIGN, TIME AND SETTING: The cell observational experiment was performed at the Central Laboratory of Daqing Oil Field General Hospital from December 2006 to December 2007.PARTICIPANTS: Dental pulp were obtained from exfoliated deciduous molar tooth of eight children aged from eight to ten, four males, four females. The possibility of infectious diseases, endocrine disease, dental caries and periodontal disease was excluded.METHODS: Deciduous teeth pulp stromal cells were obtained by tissue block adhesion method, and then cultured in DMEM with 10% fetal bovine serum for 14 days. Following digestion and passage, cells were incubated in DMEM/F12 supplemented with 50 pmol/L laevo-ascorbic acid, 10 nmol/L vitamin D3. 10 mmol/L Na β-glycerophosphate and 10-8 mol/L dexamethasone for another 14 days. Cells cultured in complete DMEM served as controls.MAIN OUTCOME MEASURES: Cell morphology and growth rule were observed under a microscope. Difference in dentin sialophesphoproteln expression was determined prior to and following mineralization using immunocytochemical staining.RESULTS: Dental pulp stromal cells from human exfoliated deciduous tooth, adhered to the wall, were polygon-shaped and characterized by large cell size and a relatively large nucleus and plenty cytoplasm after mineralization for 14 days. Over 80% cells were positive for dentin sialophosphoprotein. Non-induced cells were spindle, and only 2% cells were positive for dentin sialophosphoprotein.CONCLUSION: Odontogenic inductive culture might improve odontoblastic differentiation. It seems that human exfoliatod deciduous tooth pulp represents an new reservoir of adult stem cells with odontogenic potential.
2.Optimization of hydroxylating DHEA to 7alpha,15alpha-diOH-DHEA by compound mutation and fermentation optimization.
Chuanpeng LI ; Hui LI ; Yan WU ; Heng LI ; Rujin ZHANG ; Zhengbin ZHANG ; Jinsong SHI ; Zhenghong XU
Chinese Journal of Biotechnology 2014;30(1):147-156
Combined with method of ketoconazole resistance screening, a 7alpha,15alpha-diOH-DHEA high-producing mutant Colletotrichum lini ST-1 was obtained by compound mutation of NTG and low energy N+ ion beam implantation. With the substrate concentration of 10 g/L DHEA, the molar yield of 7alpha,15alpha-diOH-DHEA reached 34.2%, increased by 46.2% than that of the original strain. Then we optimized the medium. First, Plackett-Burman design was used to evaluate the effects of medium components on molar yield of the product. Results show that glucose, yeast extract and MgSO4 x 7H2O were the important parameters for the biotransformation process. Subsequently, the path of steepest ascent was used to approach the optimal levels. To obtain the optimal levels, central composite design and response surface analysis were carried out. The optimal medium was as follows (g/L): glucose 26.34, yeast extract 12.15, corn flour 3.00, FeSO4 x 7H2O 0.015, MgSO4 x 7H2O 0.14, KH2PO4 0.90. Under the optimal conditions, the molar yield of 7alpha,15alpha-diOH-DHEA reached 49.3%, which was 44.2% higher than that of using the medium before optimization.
Biotransformation
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Colletotrichum
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metabolism
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Dehydroepiandrosterone
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chemistry
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Fermentation
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Hydroxylation
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Industrial Microbiology
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Mutation
3.Evaluation of color Doppler ultrasound of renal blood flow combined with the detection of bone morphogenetic protein-7 in early diagnosis of type 2 diabetic nephropathy
Zhijie ZHANG ; Ning YU ; Zhengbin WANG ; Zhimei YAN ; Dongmei MENG ; Ronggui LIU ; Zhaoyan DING ; Fengfeng SHI
Chinese Journal of Ultrasonography 2012;(7):591-594
Objective To evaluate the significance of color Doppler ultrasound examination of renal blood flow combined with the detection of bone morphogenetic protein-7(BMP-7)in early diagnosis of type 2 diabetic nephropathy.Methods Blood BMP-7 level was tested in 90 patients with type 2 diabetic nephropathy and 30 controls,and parameters of renal blood flow were measured by color Doppler ultrasound examination.Blood BMP-7 level as well as resistant index(RI)of segmental renal artery(SRA)and interlobar renal artery(IRA),were compared between these two groups.Results Compared with controls,blood BMP-7 level gradually decreased with the aggravation of diabetic kidney damage(P<0.01).The peak systolic velocity(Vmax)and the end diastolic velocity(Vmin)of SRA and TRA were slowed gradually,while RI increased(P<0.01).Blood BMP-7 level was negatively correlated with IRA's and SRA's RI of IRA and SRA(r =-0.603,P<0.01;r =-0.652,P<0.01).Conclusions Color Doppler ultrasound examination of renal blood flow combined with detection of BMP-7 might play an important role in early diagnosis of type 2 diabetic nephropathy.
4.The value of virtual touch tissue quantification in the diagnosis of pancreas early damage of patients with hyperuricemia
Yunping LI ; Zhaoyan DING ; Zhengbin WANG ; Fei TIAN ; Ronggui LIU ; Guijun ZHANG ; Cheng ZHAO ; Zhimei YAN
Chinese Journal of Ultrasonography 2015;(4):316-319
Objective To explore the value of virtual touch tissue quantification (VTQ ) in the diagnosis of pancreas early damage of patients with hyperuricemia(HUA) .Methods Sixty‐five cases of patients with normal glucose tolerance and HUA (HUA group) and 42 cases of patients with HUA and impaired glucose tolerance (IGT group) were included in the study ,and other 150 health subjects were taken as control group .After routine ultrasound examination ,the pancreas shear wave velocity values (Vs) of different groups were measured by VTQ technology ,and the results were analyzed .Results Compared with the control group ,the rate of the abnormai ultrasonogram of pancreas in IGT group showed obvious increasing trend but there was no significantly difference between the other groups(both P >0 0.5) .The Vs of the normal control group ,HUA group ,and IGT group were (1 2.1 ± 0 1.5)m/s ,(1 4.1 ± 0 1.3)m/s and (1 5.9 ± 0 1.5)m/s ,respectively .The Vs of HUA group and IGT group were higher than that of the control group (both P <0 0.5) ,and the Vs of IGT group was significantly higher than that of HUA group ( P <0 0.5) .Conclusions Hyperuricemia may cause pancreas damage ,and with the disease progress ,the pancreas damage became more serious ,and VTQ technology may play an important role on the diagnosis of pancreas early damage in HUA patients .
5.Quantitative evaluation of carotid elasticity in patients with hyperuricemia by echo-tracking technology
Wen ZHAO ; Zhimei YAN ; Guijun ZHANG ; Zhengbin WANG ; Zhaoyan DING ; Yunping LI
Chinese Journal of Ultrasonography 2014;23(3):211-213
Objective To investigate the clinical value of echo-tracking(ET) technique in evaluating the carotid elasticity in patients with hyperuricemia and hyperuricemia combined with hyperlipidemia.Methods One hundred and twenty patients with hyperuricemia were divided into two group:group of hyperuricemia (group A) and group of hyperuricemia combined with hyperlipidemia (group B).Sixty normal persons were served as the control group.ET was used to measure the carotid artery elastic modulus including stiffness parameters (β),pressure-strain elastic modulus (Ep),arterial compliance (AC),augmentation index (AI) and pulse wave conducting velocity (PWVβ).Then the statistical data were analyzed to observe the changes of each parameter.Results In the comparison of three groups,elastic function index β,Ep,PWVβ and AI in group A and group B were higher than those of control group (P <0.05) ;however,AC was lower than that of control group (P < 0.05).Compared with group A,elastic function index β,Ep,PWVβ,AI was higher in group B,while the AC was lower than that of group B (P <0.05).Conclusions ET may be helpful to prevent atherosclerotic changes and to provide the basis for the clinical diagonosis and treatment atherosclerotic changes.
6.Comparison of periodontal indices and Porphyromonas gingivalis between conventional and self-ligating brackets.
Jing SHI ; Yuxin LIU ; Jingqiu HOU ; Zhengbin YAN ; Hui PENG ; Xing CHANG
West China Journal of Stomatology 2013;31(3):228-231
OBJECTIVETo compare the periodontal indices and Porphyromonas gingivalis (P. gingivalis) between the use of self-ligating brackets and conventional brackets.
METHODSThirty patients were divided into 2 groups(n=15). Self-ligating brackets were used in the experimental group. Conventional brackets were used in the control group. Clinical periodontal indices, including plaque index (PLI), gingival index (GI) and probing depth (PD) of observed teeth were examined at three different time points: Before orthodontic treatment, the first month after treatment and the third month after treatment. Subgingival plaques were collected simultaneously at each time point. The number of total bacteria and P. gingivalis in each sample were detectd and quantitated by real-time quantitative polymerase chain reaction, the percentage of P. gingivalis in total bacteria was obtained.
RESULTSBefore treatment, the periodontal indices and the percentage of P. gingivalis in total bacteria had no difference between the two groups (P>0.05). After 1 and 3 months respectively, the periodontal indices and the percentage of P. gingivalis in total bacteria increased with time (P<0.05) and were obviously lower than those of the control group (P<0.05).
CONCLUSIONCompared with conventional brackets, the self-ligating brackets are better for periodontal health. But it is adverse effect on oral health.
Dental Plaque ; Dental Plaque Index ; Humans ; Periodontal Index ; Porphyromonas gingivalis
7.Cultivation and morphological characteristics of rat adipose tissue-derived vascular endothelial cells in vitro.
Yunfeng LIN ; Xizhe CHEN ; Weidong TIAN ; Zhengbin YAN ; Xiaohui ZHENG
Journal of Biomedical Engineering 2006;23(4):836-838
The subcutaneous adipose tissue from the inguen of four Sprague-Dawley rats was obtained, then digested with one volume of collagenase type I and cultured with BGJb medium. The obtained adipose stromal cells were induced in human endothelial-SFM for 7 d. The cells were observed under inverted microscope every day and identified by transmission electron microscope and immunocytochemical staining with factor VIII antigen. The results showed the induced cells uniformly had characteristic cobblestone morphology of endothelial cells. Factor VIII antigen staining was positive in cytoplasm. Under transmission electron microscope, the cells displayed many finger like microvilli and numerous lysosomes, mitochondria, a few coarse endoplasmic reticulum and Weibel-Palade bodies. The characteristics of the rat adipose tissue-derived endothelial cells were consistent with those of vascular endothelial cells derived from other tissues. It seems that subcutaneous adipose tissue may represent a new alternative source of endogenous vascular endothelial cells.
Adipose Tissue
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cytology
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Animals
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Cell Culture Techniques
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methods
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Cell Differentiation
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Endothelial Cells
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cytology
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Endothelium, Vascular
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cytology
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Male
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Rats
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Rats, Sprague-Dawley
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Stromal Cells
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cytology