1.Protective effect of adenosine on temporary threshold shift of audition caused by intensive white noise in guinea pigs
Lan ZHANG ; Wenyuan GAO ;
Academic Journal of Second Military Medical University 2000;0(07):-
Objective: To investigate the protective effect of adenosine on hearing in acute acoustic trauma and its modulatory action in the cochlea. Methods:Guinea pigs were exposed to 118 dB SPL white noise for 30 min. During the process of exposure, artificial perilymph or artificial perilymph containing 1 mmol/L adenosine was infused into the cochlea through perilymphatic perfusion. Thresholds of compound action potentials (CAP) evoked by click and tone burst were measured before and after noise exposure. Results: Immediately after exposure, the mean threshold shifts evoked by click of the control group and adenosine treated group were (40?6.68) dB and (20?5.70) dB respectively. The threshold shifts evoked by tone burst at 2 8 kHz in the control animals were 32 50 dB, while in the adenosine treated animals, 20 35 dB. There were significant differences between the 2 groups except that at 6 kHz. Two hours after exposure, there was an increase of 4 8 dB at all frequencies in the control group; no changes in the adenosine treated group were found. There were significant differences in threshold shifts between the 2 groups at all test frequencies. Conclusion: Adenosine has protective effect on temporary threshold shift in acute acoustic trauma in guinea pigs. [
2.The strategy of gene mapping and the approach to genome-wide search for human genetic diseases
Anping ZHANG ; Xuejun ZHANG ; Wenyuan ZHU
Chinese Journal of Disease Control & Prevention 2001;5(2):135-138
The great majority of human diseases are directly or i nd irectly associated with genes. Gene mapping and genetic analysis for human compl ex polygenic disorders has become a hot-spot and the neck of the bottle in the medical genetics research recently. The approach to genome-wide search has pla yed an important role in the respect.
3.Detection of Gene Mutation in a Pedigree with X-linked Anhidrotic Ectodermal Dysplasia by PCR-SSCP Analysis
Anping ZHANG ; Xuejun ZHANG ; Wenyuan ZHU ;
Chinese Journal of Dermatology 1994;0(05):-
Objective To identify the gene mutations and mu tating patterns in a pedigree with X-linked anhidrotic ectodermal dysplasia(EDA)so as to provide a basis for gene diagn osis and genetic counselling of this disorder.Methods Polymerase chain reaction-single s trand conformation polymorphism(PCR-SSCP)analysis and DNA sequencing of amplified prod ucts were performed to screen mutati ons and mutating patterns of EDA1gen e,responsible for EDA pathogenesis,i n a X-linked EDA family of Han people.Results Abnormal single strand bands were found in the amplified fra gments as well of exon 1of EDAgene in t he patients as well as their mothers,the carriers.The DNAsequencing of ampl ified products revealed a point muta tion at nucleotide 404(C→Gtransversion)in the proband compared with that of t he normal controls,which resulted i n the transversion of histidine with glutamine at codon 54in the ectodysplasin-A(H54Q).Meanwhile there were heterozyous double peaks of nucleotide Cand Gat the same position in his mother.Conclusion A missense mutation(404C→G)in exon 1of EDA1gene has been determined in the pedigree with X-linked EDA,which is probably one of the molecular bases of EDA pathogenesis.
4.Clinical outcome of cervical arthroplasty in the treatment of spondylotic myelopathy
Wei ZHANG ; Yong SHEN ; Wenyuan DING
Orthopedic Journal of China 2006;0(15):-
[Objective]To verify the clinical outcome of cervical arthroplasty in the treatment of spondylotlc myelopathy exclusively.[Method]From January 2004 to August 2003,twelve spondylotic myelopathy patients who underwent cervical arthroplasty were included in this study.Preoperative and final follow-up Nuriek grade,Oswestry neck disability index(ONDI)score,neck and arm pain VAS score of patients were recorded and compared.Surgical outcome was also measured using odom' s criteria.[Result]100% of patients had good or excellent outcomes using Odom' s criteria,while the final follow-up Nurick grade,ONDI score,neck and arm pain VAS score were improved significantly compared with the preoperative valule.[Conclusion]Cervical arthroplasty is effective in the treatment of spinal cord compression secondary to spondylotic disease with interveterbral movement being preserved.
5.Development and clinical evaluation of iliac bars lever reduction and fixation system for spondylolisthesis
Guochuan ZHANG ; Yong SHEN ; Wenyuan DING
Orthopedic Journal of China 2006;0(04):-
[Objective]To introduce the composion and mechanism of Iliac Bars Lever Reduction and Fixation System(IBRFS),and to evaluate the clinical efficacy of IBRFS.[Method]IBRFS was made of stainless steel composing of pedicle screw,anglar lift,reduction rod,iliac bar and screw nut A,B,C.Eighteen volunteers offered their contribution for this study.There were 6 males and 12 females with mean age of 53 years(range from 43-66 years).The classification was DS 9,IS 8,trauma 1.The slip segments were L4 6,L5 12.The reductive operation was conducted by IBRFS.Clinical efficiency was evaluated by slip ratio,slip angle,sacral slope and the height of intervertebral space.[Result]Eighteen patients were followed up from 8 to 28 months(average 14.9 months).Preoperative JOA scores ranged from-1 to 22(mean 10.7).Postoperative JOA scores ranged from 13 to 29(mean26.9).The overall recovery rate ranged from 42.9% to 98%(mean 88.3%).There was no neurologic complication.Slip ratio was improved from 20.80% to 5.01%,slip angle from 6.67?to 12? and sacral slope from 32.2? to 43.3?.The height ratio of intervertebral space was increased from 0.78 to 0.97.[Conclusion]The reduction efficiency of IBRFS is reliable.The restoration of height of intervertebral space is improved.IBRFS is an effective internal fixation.
8.Three-dimensional degradable porous scaffolds made by silk fibroin-chitosan composite
Yadong YANG ; Wenyuan ZHANG ; Guojian FANG
Chinese Journal of Tissue Engineering Research 2009;13(51):10074-10078
BACKGROUND: Simply used natural materials-prepared scaffolds such as collagen, gelatin and fibrin solve problems of biocompatibility, but its degradation is rapid, and cannot induce new tissues, but collapse is found as cell scaffolds.OBJECTIVE: To explore and determine the property of biological degradable three-dimensional porous scaffolds using silk fibroin-chitosan composite.DESIGN, TIME AND SETTING: The material observational study was performed at the Institute of Bioengineering, Zhejiang Academy of Medical Science from June 2008 to June 2009.MATERIALS: Spring silk cocoon was presented by a silkworm farmer from Huangdunmiao village, Maqiao town, Haining City,Zhejiang Province, China. Chitosan was produced by Shanghai Bo'ao Biological Technology.METHODS: 15 g/L silk fibroin solution was made by degumming, salvation and dialysis. Chitosan was dissolved in 2% acetic acid solution to prepare 25 g/L chitosan-acetic acid solution. Two solutions were mixed to prepare six silk fibroin/chitosan solutions, and mass ratio was 10: 0, 5: 5, 4: 6, 3: 7, 2: 8, 0:10. These solutions were separately sucked in a 24-well plate. Following exhausting gas vacuole at 4 ℃, precooling was performed at -20 ℃ for 12 hours, followed by cryodesiccate for 30 hours. Samples were then hydrated in ethanol, neutralized in NaOH-alcohol for 1 hour, washed and then frozen to dry.MAIN OUTCOME MEASURES: Optical microscope and scanning electron microscope were used to observe pore size and structure of various mass ratio-prepared scaffold. Modified liquid substitution method was utilized to measure porosity of various scaffolds. The degradable rate of various scaffolds was determined at 4 weeks in vitro.RESULTS: Silk fibroin/chitosan of 10: 0 mass ratio-prepared scaffold had rough fluffy pore, was brittle, with high dissolve-loss rates. On the contrary, chitosan-prepared scaffold was hard, without enough elasticity following freeze-dry. The composite scaffold of 5: 5, 4: 6, 3: 7 and 2: 8 following freeze-dry was loose and soft, similar to sponge. With increased chitosan concentration,scaffold hardness increased. There were evenly distributed, detailed eyelets on the scaffold. Under the optical microscope,various pores were irregular; each pore closely connected and linked together; pore size was even, 20-100 μm. With increased chitosan concentration, pore size was gradually reduced. Scaffold porosity determination results displayed that mass ratio of silk fibroin/chitosan 4: 6 group > 5: 5 group > 3: 7 group > 2: 8 group. Compared with 2: 8 group, the porosity was significantly increased in the 5: 5 and 4: 6 groups (P < 0.05). No significant difference was detected in volume expansibility in the silk fibroin/chitosan composite scaffold of various mass ratios (P > 0.05). The degradation was slowest in the 2: 8 group, and fastest in the 5: 5 group at 4 weeks.CONCLUSION: Regarding physical and chemical properties, composite scaffold made by silk fibroin/chitosan showed significant superiority compared with scaffold made by silk fibroin or chitosan alone. Of them, silk fibroin/chitosan mass ratio of 5: 5 and 4: 6 are accorded with the requirement of cartilage tissue engineering.
9.Preparation of chitosan sustained release microspheres encapsulated with transforming growth factor beta 1
Yadong YANG ; Wenyuan ZHANG ; Guojian FANG
Chinese Journal of Tissue Engineering Research 2009;13(38):7481-7484
OBJECTIVE: To prepare chitosan microspheres encapsulated transforming growth factor β1 (TGFβ1), and to analyze its property. METHODS: The chitosan was dissolved in 2% acetic acid to prepare chitosan microspheres encapsulated TGFβ1 with emulsification cross-linking method, Tween 80 and sodium polyphosphate were served as emulsifying agent and cross-linking agent, respectively. Meanwhile, chitosan microspheres and containing bovine serum albumin chitosan microspheres were prepared as blank control and experimental control groups. The morphology and diameter of 3 kinds of microspheres were observed, and the dispersion and in vitro release of chitosan microspheres encapsulated TGFβ1 were detected, furthermore, the water absorption expansion rate of blank control and experimental control groups were measured. RESULTS: Scanning electron microscopy showed that the microspheres diameter in the blank group was approach 15 μm, with smooth surface and plenty of tiny pores. However, the microspheres in the other 2 groups were distributed uniformly with approximately 1 μm in diameter, the surface was smooth. The chitosan microspheres encapsulated TGFβ1 released fast at begin 12 hours, and then gentled gradually, with 53.5% release ratio within 6 days. The increased mass of microspheres in the blank control and experimental control groups reached a balance after 1 hour, both of which were over 700%,in particular larger in the acid environment. CONCLUSION: Chitosan microspheres encapsulated TGFβ1 prepared by emulsification cross-linking method exhibit high yield and good drug release. The strong water absorption expansion rate of chitosan microspheres requires aperture size, as well as intensity of bone tissue engineered scaffold.
10.Expression and identification of cysteine proteinase from adult worm of Pagumogonimus skrjabini in Escherichia coli
Wenyuan DUAN ; Xilin ZHANG ; Ying WANG ;
Journal of Third Military Medical University 2003;0(08):-
Objective To express the recombinant antigen coded by Pagumogonimus skrjabini adult cysteine protease gene fragment and to investigate the immunoreactivity of the fusion protein for further usage on serodiagnosis of the fluke disease. Methods The target gene fragment was amplified by PCR. After purification with gel purification recoverykit, the target gene fragment was ligated with PinPointTM Xa 1 T vector and transduced into E.coli JM109 strain. The expressed fusion protein sample was prepared with alkaline lysis solution, and then analyzed by SDS PAGE. The expression level was determined by Coomassie blue staining and the streptavidin alkaline phosphatase staining. The immunoreactivity of fusion protein was examined by Western blotting. Results A total of 8 positive clones were harvested, and only one had the proper orientation verified by sequencing. The recombinant antigen was obtained after being induced with IPTG (Isopropltio ? D galactoside), and a positive band of 32?10 3 was found by streptavidin alkaline phosphatase staining. In the same position, the fusion protein was also detected by Western blotting. Conclusion The expression vector of adult cysteine protease gene PinPointTM Xa 1 T vector was successfully constructed. The recombinant antigen obtained after being induced with IPTG possesses good immunoreactivity.