1.Preparation and properties of recombinant human bone morphogenetic protein-2/bone repair material
Chinese Journal of Tissue Engineering Research 2016;20(38):5664-5671
BACKGROUND:It has become a hotspot to prepare the bone repair material that exhibits natural bone structure and is used in combination with biological factors. OBJECTIVE:To prepare the recombinant human bone morphogenetic protein-2 (rhBMP-2)/bone repair material, and to evaluate its capacities of release, activity and ectopic osteoinduction. METHODS:A collagen-binding domain was added to the N-terminal of native rhBMP-2 that allowed bind to collagens in the bone repair material. Then, rhBMP-2/bone repair material was obtained through freeze-dried method. The releasing ability of rhBMP-2 in vitro was assayed by ELISA. C2C12 cell lines were loaded to the composite material with 0.25, 0.5 and 1 μg rhBMP-2, respectively. Afterwards, alkaline phosphatase activity was detected at 72 hours. The composite materials with 0, 2, 5 and 10 μg rhBMP-2 were implanted into the quadriceps of Sprague-Dawley rats, respectively. Alkaline phosphatase activity and the newly formed bone were detected at 2 and 4 weeks after implantation. The CY-7-labeled composite material was implanted into the quadriceps of Sprague-Dawley rats to observe its stability. RESULTS AND CONCLUSION:Substantial y no rhBMP-2 from the rhBMP-2/bone repair material was released within 45 days. The alkaline phosphatase activity of C2C12 was in a rise with the increased concentration of rhBMP-2. The stability of the composite material in vivo was better, the alkaline phosphatase activity and ectopic bone formation increased as the concentration of rhBMP-2 rose. To conclude, the rhBMP-2/bone repair material preserves the stability of rhBMP-2, and improves ectopic osteoinduction ability.
2.Changes of cAMP and cGMP contents in the central nervous system in morphine dependent and withdrawal rats
Jianping GU ; Bin CONG ; Donggang LIU
Chinese Journal of Forensic Medicine 2002;0(05):-
Objective To investigate the contents of cAMP and cGMP in the central nervous system in morphine dependent and withdrawal rats.Methods A physical morphine dependent model in rats was established by subcutaneous injection of morphine in gradually increasing doses.The cAMP and cGMP contents in the brain regions were determined by radioimmunoassay.Results Compared with control group,morphine dependence could significantly decrease the cGMP content or profoundly increase the cAMP content and the cAMP/cGMP ratio in rat striatum,diencephalons,midbrain,pons and hippocampus,but these changes described above were not detected in cerebellum.Compared with morphine dependent group,naloxone induced withdrawal could significantly decrease the cGMP contents or increase the cAMP contents and the cAMP/cGMP ratio in striatum and hippocampus,but these changes described above were not observed in the other regions.Conclusion The changes of the cAMP and cGMP contents may be one of the important molecular mechanisms leading to morphine dependence and abstinence.
4.Cloning, expression and identification of IL-1ra-Fcepsilon fusion gene.
Zhongcheng LIU ; Minji ZOU ; Yuanyuan WANG ; Jiaxi WANG ; Donggang XU
Chinese Journal of Biotechnology 2008;24(10):1754-1760
Both interleukin-1 and IgE are important in the pathogenic mechanism of the allergy asthma. cDNA of interleukin-1receptor antagonist (IL-1ra) and IgE were cloned and a prokaryotic expression vector IL-1ra-Fcepsilon/pBV220 was constructed. The vector was transformed into Escherichia coli BL21(DE3). The fusion protein was expressed successfully in the form of inclusion body. The recombination protein of IL-1ra-Fcepsilon was highly purified by chromatography of gel filtration and ion exchange, which was identifited by Western blotting. The cell assay showed that the activity of IL-1ra-Fcepsilon was as high as IL-1ra in vitro after refolding. The pharmacokenetic profile of IL-1ra-Fcepsilon and L-1ra was analyzed, and the half time of IL-1ra-Fcepsilon is 4.78 times than that of IL-1ra.
Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Gene Fusion
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Immunoglobulin E
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genetics
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metabolism
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Immunoglobulin Fc Fragments
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genetics
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metabolism
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Interleukin 1 Receptor Antagonist Protein
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genetics
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metabolism
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Recombinant Fusion Proteins
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genetics
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metabolism
5.Treating maxillary dental arch crowding and protrusion of the patient with mandibular molars missing by implanting miniscrew anchorage.
Shi-tong JIANG ; Xiao-lei JIANG ; Yong QIN ; Ru ZHANG ; Hong-jie LIU ; Guang-jun JIAO ; Kuo YUAN
West China Journal of Stomatology 2007;25(3):260-262
OBJECTIVETo study the clinical effectiveness of implanting miniscrew serving as anchorage instead of molars in treating maxillary dental arch crowding and protrusive patients mandibular molars complete missing.
METHODSEight adult patients aged from 22 to 38, whose maxillary dental arch were crowding and protrusion, with mandibular molars missing were chosed. At the missing side, a miniscrew was implanted on the buccal surface, 11-13 mm away from the distal end of the second premolar. After implanting, a self-made miniscrew traction cap was attached to its supergingival section by keyway retention. Premolars and anterior teeth were tracted in turn to distal end. Intermaxillary traction II was made necessarily on the traction cap.
RESULTSSuccessful results were acquired after treating together with normal overbite, overjet and right occlusion relation. It took 24 months in the longest course, 15 months in the shortest course and 20.8 months on average.
CONCLUSIONThe implanting miniscrew anchorages could be used in maxillary dental arch crowding and protrusive patients with single molars complete missing.
Adult ; Bicuspid ; Bone Screws ; Cephalometry ; Dental Arch ; Humans ; Molar ; Orthodontic Anchorage Procedures ; Overbite ; Tooth Movement Techniques
6.Clinical observation on therapeutic effect of suspended moxibustion on temperature-sensitive acupoints plus medication for chronic prostatitis.
Han-Shan LIU ; Ainiwan ; Reheman ; Yong FU ; Ming-Fei KANG
Chinese Acupuncture & Moxibustion 2009;29(7):543-546
OBJECTIVEObserve therapeutic effect of suspended moxibustion on temperature-sensitive acupoints for chronic prostatitis, to explore a new therapy.
METHODSSixty cases were randomly divided into an observation group and a control group (n = 30 in each group). The suspended moxibustion on temperature-sensitive acupoints plus herbalenema were practiced in the observation group. The moxibustion at Guanyuan (CV 4), Zhongji (CV 3), Shenshu (BL 23) combined with herbalenema were practiced in the control group. The score of syndromes, the score of symptom indexes (NIH-CPSI), and WBC in prostatic secretion (EPS-WBC) were used as observation index for chronic prostatitis. The therapeutic effect was compared between the two groups.
RESULTSThe markedly cured rate was 50.0% in the observation group, better than that of 26.7% in the control group (P < 0.05). The score of syndromes, NIH-CPSI and EPS-WBC in the observation group were lower than the control group (P < 0.05, P < 0.01).
CONCLUSIONThe therapeutic effect of suspended moxibustion on temperature-sensitive acupoints plus medication is better than moxibustion plus medication for chronic prostatitis in improvement of symptoms and EPS-WBC.
Acupuncture Points ; Adult ; Chronic Disease ; therapy ; Enema ; Humans ; Male ; Middle Aged ; Moxibustion ; Prostatitis ; drug therapy ; therapy ; Treatment Outcome ; Young Adult
7.Series orthodontic treatment on teeth transposition of maxillary canine and lateral incisor.
Shi-tong JIANG ; Bao-lan WANG ; Hong-jie LIU ; Guang-jun JIAO ; Zhong-jun AN ; Liang-kun JIANG
West China Journal of Stomatology 2010;28(1):74-80
OBJECTIVETo explore the treatment on teeth transposition of maxillary canine and lateral incisor in order to improve the clinical treatment effect.
METHODSEleven patients with transposition maxillary canine and lateral incisor were treated with the method: Expand space, artificial reverse occlusion of transpositional lateral incisor to give way, transpositional lateral canine distalization and controlling root, mesial movement of lateral incisor with tongue arch, interactive controlling roots and retention with tongue fixed retaining appliance.
RESULTSEleven patients had satisfactory treatment effect, with tidy dentition and parallel teeth roots of transpositional canine and lateral incisor.
CONCLUSIONSeries orthodontic treatment on teeth transposition of maxillary canine and lateral incisor can effectively improve the clinical treatment effect and shorten the treatment time.
Cuspid ; Humans ; Incisor ; Malocclusion
8.Circ_0068655 Promotes Cardiomyocyte Apoptosis via miR-498/ PAWR Axis
Qiaoying CHAI ; Mingqi ZHENG ; Zheng WANG ; Mei WE ; Yajuan YIN ; Fangfang MA ; Xinping LI ; Haijun ZHANG ; Gang LIU
Tissue Engineering and Regenerative Medicine 2020;17(5):659-670
BACKGROUND:
The cardiomyocyte apoptosis is considered as one of major contributions to cardiac remodeling after myocardial infarction (MI). Numerous studies find that circular RNAs (circRNAs) play pivotal roles in a variety of biological functions. However, the role of circ_0068655 in MI and human induced pluripotent stem-derived cardiomyocytes (HCMs) remains unknown.
METHODS:
The expression of circ_0068655, miR-498, and PRKC apoptosis WT1 regulator (PAWR) in human MI heart tissues and hypoxia subjected HCMs was evaluated with qRT-PCR and Western blot. The effects of circ_0068655 on hypoxia-induced apoptotic death and cell migration in HCMs were evaluated with qRT-PCR, cell viability, cell death ELISA (POD), and Caspase-3 activity assay, and Trans-well assay, respectively. Furthermore, luciferase assay, qRT-PCR, biotin-labeled miRNA pulldown assay, and Western blot were employed in the functional studies.
RESULTS:
We found that the expression of circ_0068655 and PAWR was enhanced in MI patients and hypoxia subjected HCMs; by contrast, the expression of miR-498 decreased. Inhibited expression of circ_0068655 in HMCs counteracted hypoxia-induced apoptotic death and impaired cell migration, in sharp contrast to circ_0068655 knockdown. We identified that circ_0068655 sponged an endogenous miR-498 to sequester and inhibit its activity, leading to the increased PAWR expression.
CONCLUSIONS
Our findings reveal that the expression of circ_0068655 can promote cardiomyocyte apoptosis through the modulation of miR-498-PAWR axis in vitro, which highlights the diagnostic and therapeutic value of circ_0068655 in patients with MI.
9.Circ_0068655 Promotes Cardiomyocyte Apoptosis via miR-498/ PAWR Axis
Qiaoying CHAI ; Mingqi ZHENG ; Zheng WANG ; Mei WE ; Yajuan YIN ; Fangfang MA ; Xinping LI ; Haijun ZHANG ; Gang LIU
Tissue Engineering and Regenerative Medicine 2020;17(5):659-670
BACKGROUND:
The cardiomyocyte apoptosis is considered as one of major contributions to cardiac remodeling after myocardial infarction (MI). Numerous studies find that circular RNAs (circRNAs) play pivotal roles in a variety of biological functions. However, the role of circ_0068655 in MI and human induced pluripotent stem-derived cardiomyocytes (HCMs) remains unknown.
METHODS:
The expression of circ_0068655, miR-498, and PRKC apoptosis WT1 regulator (PAWR) in human MI heart tissues and hypoxia subjected HCMs was evaluated with qRT-PCR and Western blot. The effects of circ_0068655 on hypoxia-induced apoptotic death and cell migration in HCMs were evaluated with qRT-PCR, cell viability, cell death ELISA (POD), and Caspase-3 activity assay, and Trans-well assay, respectively. Furthermore, luciferase assay, qRT-PCR, biotin-labeled miRNA pulldown assay, and Western blot were employed in the functional studies.
RESULTS:
We found that the expression of circ_0068655 and PAWR was enhanced in MI patients and hypoxia subjected HCMs; by contrast, the expression of miR-498 decreased. Inhibited expression of circ_0068655 in HMCs counteracted hypoxia-induced apoptotic death and impaired cell migration, in sharp contrast to circ_0068655 knockdown. We identified that circ_0068655 sponged an endogenous miR-498 to sequester and inhibit its activity, leading to the increased PAWR expression.
CONCLUSIONS
Our findings reveal that the expression of circ_0068655 can promote cardiomyocyte apoptosis through the modulation of miR-498-PAWR axis in vitro, which highlights the diagnostic and therapeutic value of circ_0068655 in patients with MI.