Adult ; Carcinoid Tumor ; pathology ; Ear Neoplasms ; pathology ; Ear, Middle ; pathology ; Female ; Humans

Objective To explore the expression pattern of tenascin-c(tnc)gene in zebrafish embryo abnormal development which was induced by ethanol,and to further understand the function of tnc gene in embryo develepment.Methods Zebrafish were treated with ethanol at different concentration from 100 to 500 mmol/L,and embryos at 24 and 48 hours were collected and fixed,then tnc expression pattern was observed by in situ hybridization and reverse transcriptase-polymerase chain reaction(RT-PCR).Results The result of RT-PCR showed that ethanol at 100 and 200 mmol/L could increase the expression of tnc,while the result of in situ hybridization showed that,while ethanol at 300 mmol/L and above decrease the expression of tnc in presumptive position at 24 hours,and ethanol at 100 mmol/L and above caused increase expression of tnc in zebrafish heart.Conclusions tnc is increased when treated with 100 and 200 mmol/L ethanol and is presented in the abnormal development of hearts of zebrafish,which can promote the normal development of embryos in some degrees.The expression pattern of tnc in pathologic state is highly conserved in all vertebrate,and in adult and embryos as well.

ObjectiveTo prepare a novel etimicin-encapsuled chitosan/hydroxyapatite nano-scaffolds and offer assistances for bone defect or osteomylitis.MethodsDrug-carried chitosan nanoparticles which was prepared by ionotropic gelation were combined with nano-hydroxyapatite.The mixture were shaped in molds and then prepared into porous scaffolds by freeze-dry.The surface of one scaffold was scanned.The grinded,particles of the scaffold were detected by field emission scanning electron microscope; X-ray diffraction was used to analyze components of the scaffold and total porosity.Staphylococcus aureus was choosed as the experimental bacteria,we studied lasting antibacterial property of drug-carried bone scaffold by antibacterial experiments,long-term drug releasing experiments and accumulation drug releasing experiments.Bone mesenchymal stem cells were used to detect the histocompatibility and inductivity of etimicin-carried scaffold.ResultsFreeze-dried porous scaffold has a surface with proper pore distribution (total porosity 70.68％) and the grinded scaffold has a globular and coliformed microstructure known after scanned by electron microscope.The drug-carried scaffold has a typical wave of hydroxyapatite under X-ray diffraction.The lasting antibacterial property study indicated that the drug-carried bone scaffold had maintained an inhibition zone for more than 7 days.The long-term drug releasing experiments and accumulation drug releasing experiments show that the fictional drug-carried bone scaffold released above the bacteriostasis concentration after one week and the accumulative amount within the safety scale.The scaffold had not an inhibitory effect on bone mesenchymal stem cells.ConclusionThe etimicin-encapsuled chitosar/ hydroxyapatite nano-scaffolds has similar microstructure and components of bone tissue.It is promising in bone tissue engineering applications because of its slow-release,antibacterial properties and satisfactory histocompatibility.

Objective To explore the temporal and spatial expression pattern of tenascin-c(tnc) and tenascin-w(tnw) in zebrafish early development,to further explore the role of tenacsin in zebrafish embryo development,and the association between them.Methods Zebrafish embryos at 2 hours post fertilization(hpf),4 hpt,8 hpt,10 hpf,24 hpt,48 hpr,72 hpf and 7 days post fertilization(dpf) were collected to extract RNA for reverse transcription-polymerase chain reaction(RT-PCR) and fix the embryos at different stages for in situ hybridization.Temporal and spatial expression pattern of tnc and tnw on different stages of zebrafish early development was observed.Results tnc and tnw all expressed in zebrafish from 24 hpf to 7 dpf,but did not expressed from 2 hpf to 10 hpf.Tnc expressed at pharyngeal arch,notochord,somite in 24 hpf,then weakly expressed at somite,but highly expressed at otic vesicle,pectoral fin and hindbrain in 48 hpf,and tnc was expressed at hindbrain,pharyngeal and notochord and disappeared at somite and pectoral.tnw expressed at hindbrain,midbrain and otic vesicle in 24 hpf,expressed at somite,notochord,hindbrain,otic vesicle and pharyngeal in 48 hpf.In 72 hpf,tnw expressed weakly at somite and notochord.Conclusions Zebrafish tnc and tnw have special temporal and spatial expression pattern,and share partial overlapping expression pattern.

Toad venom is the Bufo bufo gargarizans or B. melanostictus after the ears of the gland secretion, used in the treatment of various cancers in recent years. Research shows that the main anti-tumor components in bufadienolide. Bufadienolide have free type structure and conjunct type structure. To identify and clarify the difference between bufogenin and bufotoxin contained in Bufonis Venenum, which was from B. bufo gargarizans, an UPLC-TQ-MS method has been established. UPLC-TQ-MS method was used to identify and quantify the major bufadienolides in Bufonis Venenum. UPLC-TQ-MS assay with positive ion mode was performed on a Waters ACQUITY UPLC BEH C, (2.1 mm x 100 mm, 1.7 µm) with the mobile phase consisting of 0. 1% aqueous formic and acidacetonitrile in gradient elution at a flow rate of 0.4 mL · min⁻¹ and the column temperature was set at 35 °C. By comparing their retention time and high resolution mass data of Bufonis Venenum extracts, 37 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were free-type bufadienolides (bufogenin), 11 of them were conjugated bufadienolides. There were significant differences in the main composition between fresh and processed Bufonis Venenum. The study found that the chemical composition of toad venom through great changes after processing, conjunct type content is much less, free type content as well change.
Amphibian Venoms ; chemistry ; metabolism ; Animals ; Bufonidae ; classification ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Molecular Structure ; Tandem Mass Spectrometry ; methods

Humans ; Leukemia, Myeloid, Acute ; Mutation ; Prognosis ; Remission Induction ; fms-Like Tyrosine Kinase 3

Adult ; Aged ; Female ; Hepatitis B, Chronic ; complications ; therapy ; Humans ; Liver Failure ; etiology ; therapy ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged ; Treatment Outcome

OBJECTIVETo reproduce a reliable rat model of burn with infection for the study of prevention and treatment of infected wound.

METHODS(1) Electrical burn producing apparatus equipped with constant temperature (80°C) and pressure (0.5 kg) was used to reproduce burn injury (with area of 4.5 cm(2)) on both sides of the back in 50 SD rats for different duration (4, 6, 8, 10, 12 s), with 10 rats for each burn duration. On post burn day (PBD) 1, gross condition of wounds was observed with naked eyes. Wounds on the left side were used to observe healing time. The wounds on the right side were used for histological observation to determine the depth of injury, and they were classified into superficial and deep partial-thickness injury. (2) Another 36 SD rats were divided into A (inflicted with superficial partial-thickness burn, n = 18) and B (inflicted with deep partial-thickness burn, n = 18) groups according to the random number table. Rats in both groups were treated in accordance with method of preliminary experiment. Immediately after burn, 0.1 mL of liquid containing 1 × 10(9), 1 × 10(7), 1 × 10(5) CFU Pseudomonas aeruginosa (PA) ATCC 27853 was respectively inoculated to the wounds on one side (with 6 rats for each amount), while the wounds on the other side were treated with the same volume of normal saline as control. Inflammatory reaction of wounds was examined with HE staining on post inoculation day (PID) 1. On PID 1, 2, 3, 5, 7, and 14, the number of subeschar bacteria was respectively counted and the bacteria were identified with Gram stain and biochemical reaction. Wound healing time was recorded. Data were processed with t test.

RESULTS(1) Burn for 6, 8 s was respectively identified as injury time resulting in superficial or deep partial-thickness injury according to histological observation and wound healing time. (2) Obvious inflammatory cell infiltration was observed in the wounds in B group which were inoculated with 1 × 10(7), 1 × 10(9) CFU PA, and the infiltration was less marked in A group with inoculation of 1 × 10(9) CFU PA. (3) The bacteria isolated from wounds of A and B groups was identified as PA. The subeschar bacteria count within PID 14 in A group, in which different amount of PA was inoculated, was mostly less than 1 × 10(5) CFU/g of tissue, while that in B group in which 1 × 10(9) CFU PA was inoculated was more than 1 × 10(5) CFU/g of tissue. (4) There was no obvious difference in wound healing time between wounds inoculated with different amount of PA and wounds treated with normal saline in A group (with t value respectively 1.26, 0.29, 1.07, P values all above 0.05). Wound healing time of wounds in B group, in which 1 × 10(9) CFU PA was inoculated, was longer as compared with that treated with normal saline [(22.5 ± 1.0) d vs. (19.4 ± 1.6) d, t = 2.73, P < 0.05].

CONCLUSIONSIn rat, deep partial-thickness burn wound inoculated with 1 × 10(9) CFU PA ATCC 27853 is a reliable model with high reproducibility for the study of infection of burn wound.

Objective To prepare T-helper cell(Th)epitnpe-modified human soluble B cell activating factor belonging to the tumor necrosis factor family(BAFF)mutants and evaluate their immune response in vaccinated mice.Methods Recombinant cDNAs were cloned and ligated into the prokaryotie expression vec- tor pQE-80L respectively.The recombinant proteins were induced to express by IPTG in E.coli DH5?and purified with Ni-NTA chromatography.BALB/c mice were immunized with recombinant proteins respectively and the titres of the antibodies that were cross-reactive with BAFF in sera were analyzed by ELISA.Inhibiting ability of the antibodies in sera was analyzed by MTT assay.Results The recombinant proteins were highly expressed in E.coli DH5?.After purification,the purity of recombinant proteins was more than 90%.BALB/c mice immunized with recombinant proteins produced high levels of BAFF-specific antibodies.Cell proliferation assay showed that the sera of immunized mice could inhibit the proliferation-inducing activity of recombinant sBAFF and natural sBAFF.Conclusion The immune inhibitors of human BAFF which can induce polyclonal antibodies that are cross-reactive with BAFF are successfully prepared.These results may provide the basis for further study of their therapeutic effects.

Objective:To conduct a systematic study of the immunologic response of rats to transplanted glutaraldehyde ( GA)-treated porcine blood vessels in vivo.Methods: The experiment was divided into two groups:fresh group and glutaraldehyde-treated group.Twenty cases of fresh and glutaraldehyde-treated porcine pulmonary arteries were subcutaneously embedded in rats.We compared the changes using HE staining and immunohistochemistry.Results:HE staining showed that there were stronger expression on day 12 and day 30 in the fresh group than that in the glutaraldehyde group.There were similar results in morphology in CD68,C3,IgG.The results of integral optical density ( IOD) in immunohistochemistry showed that IOD started rising from day 4 and got the peak on day 12 or day 30 and or fell on day 60.Conclusion: Innate immunity played an important role in the research on xenogenic immunological rejection mechanism.The immunogenicity of glutaraldehyde-treated xenogenic blood vessels is lower than that in fresh blood vessels.However there is still immunogenicity in glutaraldehyde-treated xenogenic blood vessels.We will explore better ways to obviously weaken the rejection.