1.Simulated body fluid immersion method for assessing biological characteristics of calcium citrate
Xiuming DING ; Lei PENG ; Feng WEN ; Zhaowei TAN ; Zhonglin MU
Chinese Journal of Tissue Engineering Research 2013;(38):6811-6816
BACKGROUND:Calcium citrate has a better solubility than calcium phosphate, calcium sulfate, and other calcium biomaterials. The synthetic calcium citrate has a good denseness, and stably releases calcium ions at a high efficiency during the degradation. Consequently, it may be more suitable for the fil ing of fracture defects, providing needed calcium ions for early fracture healing.
OBJECTIVE:To prepare calcium citrate biomaterials with a novel formulation based on the natural bio-mineralized oyster shel s and citric acid so as to expect to get a good application in fracture healing repair.
METHODS:Crushing, grinding, and chemical reaction methods were used for refinement. Particle size analyzer, X-ray diffraction, scanning electron microscope, and Fourier transform infrared spectroscopy were adopted for analysis of the size distribution, composition, mineral phases, and micro-morphology. Biological characteristics were evaluated through a simulated body fluid experiment.
RESULTS AND CONCLUSION:Oyster shel powder was reacted with saturated citric acid to produce the calcium citrate material that had uniform crystal structure and compact bonding among crystal bodies, and exhibited a certain mechanical ability. The calcium citrate material had a good crystal structure that was conductive to prolong the degradation time. The calcium citrate released calcium ions slowly, and did not produce dramatic changes in the pH value (7.20-7.46) of the surrounding in the dissolution process. With the gradual degradation of calcium citrate materials, Ca2+concentration in solution increased gradual y and stably, and ultimately achieved an appropriate concentration of 7 mmol/L, suitable for osteoblast proliferation and differentiation. Calcium citrate prepared using natural oyster shel has good biological properties, and exhibits a natural superiority to artificial bone materials.
2.Generation and functional analysis of EBV LMP2A specific CTL elicited by DC transfected with recombinant adenovirus in vitro
Kun YAO ; Guangyong PENG ; Chuanlin DING ; Feng ZHOU ; Fangyi XIE ;
Chinese Journal of Immunology 1999;0(12):-
Objective:Generation and functional analysis of EBV LMP2A specific CTL elicited by DC transfected with recombinant adenovirus in vitro .Methods:PBMC were isolated from healthy EBV carriers and NPC patients, and then cocultured with autologous mature Ad5 LMP2A transfected DC at the ratios of 20∶1. Cytotoxicity of LMP2A specific CTL was determined with LDH release assay, the populations of CTL were performed by FACS,the IFN ? secretion and FasL mRNA expression of the CTL were detected by biological activity assays and RT PCR, respectively.Results:The results showed that high cytotoxicity of LMP2A specific CTL could be elicited by autologous transfected DC. The cytotoxicity boosted with the increase of transfected DC stimulation times, but there were no significant changes between two and three stimulations.The phenotypic analysis demonstrated that the LMP2A specific CTL induced at day 14 consisted of a majority of CD4 + and CD8 +T cells, and only a small percentage of CD56 + cells. The IFN ? secreted in the supernatants of cell culture also increased with the stimulation times. In addition, the specific CTL at day 14 from EBV healthy carriers could express FasL mRNA.Conclusion:Strong and functional EBV specific CTL could be generated by autologous mature DC transfected with adenovirus encoding LMP2A, which could provide a rationale for the immunotherapy of EBV associated NPC. [
3.Preparation and characterization of a new microporous bacterial cellulose material as a potential scaffold
Jiye DING ; Peng ZHANG ; Lin CHEN ; Feng HONG
International Journal of Biomedical Engineering 2016;39(1):10-15,后插5
Objective When bacterial cellulose (BC) is used as a scaffold material in tissue engineering,the nano-structure of BC may not provide enough space for animal cell growth and differentiation which would not achieve a perfect application in tissue engineering.In order to solve this problem,a novel green approach is developed in this research to produce bacterial nanocellulose materials with micropores ranging 50-800 μm.Methods Several ratios of hydrogen peroxide to sodium chlorite were used to react instantly to produce a large number of bubbles in BC hydrogels,which formed micropores with diameters ranging 50-800 μm.Optical microscopy and scanning electron microscope were used to evaluate microporous BC hydrogels and verify the existence of micropores.Results The size of pores could be regulated along with the changes in the amount of reactants used in the experiment.Fourier transform infrared spectroscopy verified that no cellulose was oxidized.Water content of the microporous BC hydrogels was similar to that of the original BC hydrogels.The Young's modulus of microporous BC hydrogels was 26.1 kPa,which was lower than that of the original BC hydrogels (69.9 kPa).Thiazoyl blue tetrazolium bromide (MTT) test displayed a higher viability on the microporous BC hydrogels compared to the growth on the unmodified BC substrates.Conclusions This study provides a convenient and promising way to prepare microporous materials,which may not be limited to only BC material,but could be used in other hydrogels.The proposed approach is suitable for extensive industrialization.
4.The modified antegrade digital artery island flap for severely flexion contracture of the burned finger
Shiming FENG ; Peng DING ; Aiguo WANG ; Zaiyi ZHANG ; Jian CHENG ; Qingqing SUN ; Yanyun WU
Chinese Journal of Microsurgery 2016;39(5):445-448
Objective To investigate the clinical outcomes of using modified antegrade digital artery island flap for the treatment of the severely flexion contracture of the burned finger.Methods Between August,2013 to August,2015,21 patients (21 fingers) with severely flexion contracture of the burned finger were hospitalized for treatment.According to the Stren classification standard for the interphalangeal joint flexion contracture,all the patients were rated as type Ⅲ.The volar soft-tissue defect with exposed tendons,nerves,vessels or bone ranged from 1.0 cm × 2.0 cm to 2.5 cm × 4.0 cm after scar relaxation.The artery and the nerve defect were 1.5 to 4.5 cm and 2.0 to 4.2 cm,respectively.The wound were reconstructed with the modified antegrade digital artery island flap.The dorsal branches of the proper digital nerve of the flap were anastomosised with the proper digital nerve of the wound.The flap donor site was resurfaced with full-thickness skin grafting from inner aspect of the forearm.All the cases were called back for postoperative follow-up.Results All the reconstructed fingers and flaps survived completely without vascular problems.The donor skin graft survived and wound healed by first intention.All the patients were followed up with 11.5 months (range,6-22 months).The finger appearance was satisfactory.The texture and color of flaps in all cases were good.There was no pigmentation and contraction relapse.The contracted fingers received no cold intolerance.At the final examination,the average values of static 2-point discrimination were 5.2 mm (range,4.3-6.5 mm) of the flap.In the series,based on the Michigan Hand Outcome Questionnaire,18 patients were strongly satisfied with the injured finger appearance and 3 patients satisfied with the appearance.Conclusion The modified antegrade digital artery island flap,which is easy to raise with large flap size and can result with the good finger appearance and function,is an ideal technique for reconstruction of the severe flexion contracture of the burned finger.
5.Chemokine stromal cell-derived factor-1 and its receptor CXCR4 mediate migration of marrow stromal cells into the lesion site of completely transected spinal cord
Peng DING ; Liping XUE ; Zhiyong YANG ; Chongqian WANG ; Jiahu WANG ; Zhongtang FENG ; Rongan LING
Chinese Journal of Tissue Engineering Research 2009;13(27):5380-5384
BACKGROUND: Marrow stromal cells (MSCs) own the characteristic of migration. However, the mechanisms underlying the migration of these cells remain unclear. OBJECTIVE: To explore the roles of stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 in trafficking of MSCs migration. DESIGN, TIME AND SETTING: The in vivo cytology experiment was performed at Department of Anatomy, National University of Singapore from March 2007 to June 2007. MATERIALS: MSCs were isolated and purified from a Wistar neonatal rat. Forty adult female Wistar rats were randomly divided into sham operation and experimental groups, with 20 animals in each group. METHODS: The chemotaxis assay was performed at a 48 well Boyden chamber, and a total of 25 μL SDF-1 was added to the lower layer of chamber, covered with 8 μm polycarbonate membrane filter; SDF-1 cultured in DMEM conditioned medium was served as a blank control group. Cell concentration was regulated to 1.5×109L-1/L. 50 μL and cell suspension was added into the upper layer of chamber, cultured at CO2 incubator with temperature of 37 ℃ for 10 hours. Rats in the experimental group were prepared for transected spinal cord injury models, and in the sham operation'group, only the vertebral plate was opened. 1.0 mL (1×109L-1/L) MSCs suspension labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) was injected through internal jugular vein at 1 hour after completely transected spinal cord. MAIN OUTCOME MEASURES: Expression of chemokine receptor CXCR4 in MSCs, as well as the effect of SDF-1 on the migration of MSCs was observed by immunofluorescence, change of SDF-1 in lesion site of spinal cord was detected by real-time PCR analysis, as well as the in vivo migration of intravenously injected MSCs was detected by fluorescence microscopy. RESULTS: The pudfied MSCs were positive to CXCR4. Compared to the blank control group, SDF-1 with concentrations of 5, 50, and 500 μg/L could accelerated the migration of MSCs (P < 0.05), which reached a peak with concentration of 500 μg/L. The expression of SDF-1 RNA was obvious increased in the experimental group than that of the sham operation group (P < 0.05), and returned to a normal level at 14 days. At 2 weeks after cell injection, the number of MSCs migrated to the lesion site of completely transected spinal cord was significant increased than sham operation group (P < 0.05). CONCLUSION: SDF-1 may contribute to MSCs migration in vitro and in vivo. SDF-1 and its receptor CXCR4 are involved in the migration of injected MSCs to the lesion site of completely transected spinal cord.Ding P, Xue LP, Yang ZY, Wang CQ, Wang JH, Feng ZT, Ling EA.Chemokine stromal cell-derived factor-1 and its receptor CXCR4 mediate migration of marrow stromal ceils into the lesion site of completely transected spinal cord.
6.Application value of three-dimensional female pelvic visualization modeling on transvaginal gynecological ultrasound imaging
Feng ZHANG ; Liwen TAN ; Yanli GUO ; Haiyun HUANG ; Jun DING ; Xing HUA ; Li PENG
Chinese Journal of Obstetrics and Gynecology 2014;49(2):100-103
Objective To explore the application value of three-dimensional (3D) female pelvic visualization model on transvaginal gynecological ultrasound imaging.Methods The second digital Chinese woman visible human datasets was selected,and a female pelvicvoxel-based model was constructed in Amira software for analog simulation combined with transvaginal gynecological ultrasound.The main structures and the space position variation of female pelvic cavity were observed,and the comparison study with conventional section of ultrasound was carried out.Results A high accuracy 3D visualization model of female pelvic was successfully constructed,and the anatomy structures can be dynamically and clearly displayed in any section orientation.An analog simulation research was carried out according to the scan mode of gynecological ultrasound,and a series of high resolution images consistent with scan sections of dynamic ultrasound were obtained,which had provided the optimal scan pathway for the fast inspection of the uterus,fallopian tubes and ovaries.Conclusion The 3D female pelvic visualization model based on digital Chinese woman human datasets has provided detailed and accurate anatomy information for transvaginal gynecological ultrasound imaging,which could be the benefit for doctors to understand the ultrasound images of female pelvic,as well as the accurate positioning ability to the lesions of female pelvic cavity.
7.miRNA-124 in immune system and immune disorders
QIN ZHEN ; WANG PENG-YUAN ; SU DING-FENG ; LIU XIA
Chinese Journal of Pharmacology and Toxicology 2017;31(10):998-998
In recent years, miR-124 has emerged as a critical modulator of immunity and inflammation. Here, we summarize studies on the function and mechanism of miR-124 in the immune system and immunity-related diseases. They indicated that miR-124 exerts a crucial role in the development of immune system, regulation of immune responses, and inflammatory disorders. It is evident that miR-124 may serve as an informative diagnostic biomarker and therapeutic target in the future.
8.Effect of different treatment on endophytic bacterial communities in continuous cropping of Chrysanthemum morifoliu.
San-meil PENG ; Bo-lin WANG ; Jian-zhong XU ; Zhi-shan DING ; Xiao-feng YUAN
China Journal of Chinese Materia Medica 2014;39(24):4763-4768
To reveal the effect of rotation cropping and bacterial manure on the growth of Chrysanthemum morifolium and screen the beneficial endophytic, the diversity of endophytic and dominant genera of different treatment groups were analyzed. Four different treatments were continuous cropping, rotation, self-made organic fertilizer and commercially available fertilizer, respectively. Endophytic bacterial diversity and dominant genera in different organs were examined using Terminal Restriction Fragment Length Polymorphism (T-RFLP). The results showed that enzyme Hae III was more appropriate than enzyme Hinfl because the number of TRFs digested by enzyme Hae III was more than that of enzyme Hinfl. In comparison of diversity, the endophytic bacterial communities' diversity index in group of cropping rotation and fertilizer was higher than that of continuous cropping which indicated that the addition of exogenous microorganism in soil could increase the diversity of plant endophyte. 18 dominant species were selected, including 3 kinds of Firmicutes, 4 kinds of Actinomycetes and 11 kinds of Proteobacteria. The results of dominant species comparison showed that the number of dominant species in continuous cropping of Ch. morifolium was significantly less than that of the rotation group. Some dominant bacteria in rotation group and fertilizer group such as Arthrobacter, Streptomyces, Streptomyces, Flavobacterium and Mycobacterium were not found in the continuous cropping of Ch. mortfolium group. Dominant species of fertilizer treatment group was similar with the rotation group, and the continuous cropping group's dominant species was more abundant. It indicates that these bacteria may be able to mitigate hindrance in continuous cropping, especially the Flavobacterium which can decompose the pathogenic fungi is worthy of further attention. Compared with leaves, there are more dominant species in roots and stems. The diversity of edophytic bacterial communities in continuous cropping of Ch. morifolium stays below than that in the rotation of Ch. morifolium, and fertilizer treatment can increase the diversity of continuous cropping so that it could mitigate hindrance in continuous cropping.
Actinobacteria
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physiology
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Agriculture
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Biodiversity
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Chrysanthemum
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growth & development
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microbiology
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Deoxyribonucleases, Type II Site-Specific
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Endophytes
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Fertilizers
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Gram-Positive Bacteria
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physiology
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Phylogeny
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Plant Leaves
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Plant Roots
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microbiology
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Polymorphism, Restriction Fragment Length
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Proteobacteria
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physiology
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RNA, Ribosomal, 16S
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chemistry
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genetics
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Soil
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Soil Microbiology
9.Differences of fungal diversity and structure in rhizosphere of Fritillaria thunbergii from different provenances.
Xiao-feng YUAN ; San-mei PENG ; Bo-lin WANG ; Zhi-shan DING
China Journal of Chinese Materia Medica 2014;39(22):4304-4310
To explore the mechanism of soil microbial ecology, the differences of fungal diversities in rhizosphere of different provenances of Fritillaria thunbergii were analyzed. The diversities and compositions of rhizo-fungi of the samples were analyzed by using DGGE and 454 pyrosequencing. DGGE results showed the Shannon index of Ninbo provenance planted in Ninbo was the highest one. And its dominant fungi were Ascomycota, Deuteromycota and Zygomycota. Except the same fungi, every provenance planted in Ninbo had its own special ones. From the 454 pyrosequencing, the fungal diversity in Panan producing was the highest which was similar with DGGE result. Among the ten phylum detected in its rhizosoil, Fungi_incertae_sedis, Ascomycota, Mucoromycotina, Basidiomycota and Chytridiomycota almost amounted to 90% of the whole community. The fungal types and amounts in Panan were more than those in Ninbo indicating the differences between producing areas and the advantage of macro genome sequencing. There were 10 phyla, 29 families, 28 genus and 159 species of fungi in Panan provenance, 6 phyla, 20 families, 19 genus, 136 species in Ninbo provenance, 8 phyla, 37 families, 47 genus, 289 species in Nantong provenance and 7 phyla, 25 families, 24 genus, 102 species in the bulk soil. Some genus such as Dothidea, Capnobotryella and Conidiobolus were only existed in Nantong provenance, while Pyrenochae- ta, Glomus and Pseudonectria were only in Panan provenance, which implied these species could grew because F. thunbergii influenced the existence of fungi. Experiments of provenance and producing area of F. thunbergii showed that the fungal diversity of indigenous provenance was higher than that of exotic provenance and each provenance had unique fungal species in the rhizosphere, which indicated that the diversity and structure was shaped cooperatively by the species and soil type. These fungal species are interacted with the soil-rhizhosphere-microbe microecological system, which in turn influence the growth of F. thunbergii.
Ecosystem
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Fritillaria
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genetics
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microbiology
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Fungi
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genetics
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Rhizosphere
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Soil
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Soil Microbiology
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Species Specificity
10.Effects of growth years of Paeonia lactiflora on bacterial community in rhizosphere soil and paeoniflorin content.
Xiao-Feng YUAN ; San-Mei PENG ; Bo-Lin WANG ; Zhi-Shan DING
China Journal of Chinese Materia Medica 2014;39(15):2886-2892
To explore the relationship between microecological environment and Paeonia lactiflora the effects of growth years of P. lactillora on rhizosphere bacterial communities were studied by PCR-DGGE and the paeoniflorin content determined by HPLC. Results showed that the soil pH increased with growing years of P. lactillora. In the fourth year, soil pH and enzyme activity reached the highest level, while organic matter content was the lowest. The bacterial diversity had a positive correlation with growing years varied from 3.38 to 3.61. Sequencing results demonstrated that Gammaproteobacteria, llphaproteobacteria, Actinobacteria, Acidobacte- ria and Firmicutes were predominant bacteria kinds in the soil of P. lactillora. Gammaproteobacteria was only detected in the bulk soil, while llphaproteobacteria, Acidobacteria G1l, Actinobacteria were only in the rhizosphere soil and the bacterial community among different growing years were similar except few species. HLPC results showed that paeoniflorin content was 3.26%, 3.30%, 3.36%, 3.41% separately from one to four-year-old P. lactiflora with an upward trend. The correlation analysis indicated that the paeoniflorin content had a positive correlation with soil pH and bacterial diversity, conversely, had a negative correlation with organic matter con- tent. During the growth years the rhizosphere bacterial diversity increased without changes of predominant bacteria and the paeoniflorin content increased without significant differences while its production increased significantly, which was different from the plants showing replanting diseases. This is in line with the farming practice choosing 4-year-old P. lactllora, but not the 1-3 year old one. In addition, the accumulation of paeoniflorin is closely related to soil pH, organic matter content and bacteria diversity, confirming that the geoherblism of P. lactiflora is closely related with microbial environment in the soil.
Bacteria
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classification
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Biodiversity
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Glucosides
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metabolism
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Hydrogen-Ion Concentration
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Monoterpenes
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metabolism
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Paeonia
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growth & development
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metabolism
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microbiology
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Rhizosphere
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Soil
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Soil Microbiology
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Temperature