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.

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.

Objective To investigate the clinical application of the combined the anterior malleolus flap and anterior tibia flap. Methods Based on the dissection of the perforating branches of anterior tibial artery on the middle and inferior section, the combined the anterior malleolus flap and anterior tibia flap was designed to repair the necrotic skin of anterior foot for 5 patients. The sizes of the flaps ranged from 17 cm×10 cm-10 cm× 5 cm. And the area of the flap was from tibial tuberosity(upper bound) to the line between internal malleolus and external malleolus (lower bound), and from the median line of one side of leg to the other side. Results Postoperatively, all flaps survived, and the primary healing of transplanted skin in donor site was achieved. The texture of flaps were excellent, the phenomenon of abrasion did not happen, and the clinical therapeutic efficacy was satisfactory after a follow up of 2-24 months. Conclusion It's a good method that combined the anterior malleohls flap and anterior tibia flap, which not only could enlarge the area of the flap but also has reliable blood supply, in repair of large size skin defect of anterior foot.

Objective To analyze the results of HBsAg weakly positive and both HBsAg and HBsAb simultaneously positive in 5 items of hepatitis B detected by ELISA .Methods 115 cases of HBsAg weakly positive and 95 cases of both HBsAg and HBsAb simultaneously positive were screened out from 35 280 cases of 5 items detection results of hepatitis B .210 screened samples were performed the electrochemiluminescence immunoassay (ECLIA) quantitation .Results 115 cases of HBsAg weakly positive were re‐detected by using ECLIA ,90 cases had the consistent results with the coincidence rate of 78 .3% .After ECLIA re‐detection in 95 cases of HBsAg and HBsAb double positive results ,11 cases had the consistent results with the coincidence rate of 11 .6% .Conclu‐sion The results of HBsAg weakly positive and both HBsAg and HBsAb double positive in 5 items of hepatitis B detected by ELISA must be cautious .In the detection results of HBsAg weakly positive ,the majority are the samples of HBsAg ,HBeAb and HBcAb positive and HBsAg and HBcAb positive .The results of HBsAg and HBsAb simultaneously positive have poor reliability , which should be careful to issue the detection reports .

Objective To investigate the expression of lymphotoxin-β and its receptor in the skin of a mouse model of allergic contact dermatitis (ACD).Methods BALB/c female mice at 6 to 8 weeks of age were randomly divided into the experiment group (n =15) and normal control group (n =10).Topical 2,4-dinitrofluorobenzene (DNFB,0.5％) was used to establish a model of ACD in the experiment group followed by challenge with DNFB (0.2％) at the right external ear on the 6th day.The normal control mice remained untreated.The right external ear was resected from all of the mice 24 hours after the challenge and divided into 2 parts for the measurement of mRNA and protein expressions of lymphotoxin-β and lymphotoxin-βreceptor by fluorescence-based real-time quantitative PCR and immunohistochemistry,respectively.Results The median mRNA expression levels of lymphotoxin-β and lymphotoxin-β receptor were 3.630 and 1.148respectively in the experimental mice,with the interquartile range being 1.187 and 0.617 respectively,and were both 1.000 in the control mice with the interquartile range being both 0.000.There was a significant difference between the experimental and control group in the mRNA expression levels of lymphotoxin-β and lymphotoxin-β receptor (P ＜ 0.01 ).Immunohistochemistry showed a significant increase in the expression of lymphotoxin-β and lymphotoxin-β receptor proteins in the experimental group compared with the control group (P ＜ 0.01 ).Conclusion There is an increment in the mRNA and protein expressions of lymphotoxin-β and lymphotoxin-β receptor in the lesions of ACD in mice.

Gliomas are one of the most common types of brain cancers. Numerous efforts have been devoted to studying the mechanisms of glioma genesis and identifying biomarkers for diagnosis and treatment. To help further investigations, we present a comprehensive database named GliomaDB. GliomaDB includes 21,086 samples from 4303 patients and integrates genomic, transcriptomic, epigenomic, clinical, and gene-drug association data regarding glioblastoma multiforme (GBM) and low-grade glioma (LGG) from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), the Chinese Glioma Genome Atlas (CGGA), the Memorial Sloan Kettering Cancer Center Integrated Mutation Profiling of Actionable Cancer Targets (MSK-IMPACT), the US Food and Drug Administration (FDA), and PharmGKB. GliomaDB offers a user-friendly interface for two main types of functionalities. The first comprises queries of (i) somatic mutations, (ii) gene expression, (iii) microRNA (miRNA) expression, and (iv) DNA methylation. In addition, queries can be executed at the gene, region, and base level. Second, GliomaDB allows users to perform survival analysis, coexpression network visualization, multi-omics data visualization, and targeted drug recommendations based on personalized variations. GliomaDB bridges the gap between glioma genomics big data and the delivery of integrated information for end users, thus enabling both researchers and clinicians to effectively use publicly available data and empowering the progression of precision medicine in glioma. GliomaDB is freely accessible at http://bigd.big.ac.cn/gliomaDB.

10.3969/j.issn.2095-4344.2013.25.016

Objective To construct recombinant adenovirus vector containing Rattus norvegicus microtubule-associated protein 4 gene,and transfect it into the rat cardiac myocytes cultured in vitro.Methods A pair of primers were designed,and full length MAP4 DNA was cloned from rat total mRNA by PCR.The PCR product was double-digested with restriction endonucleases NheⅠ and NocⅠ,and inserted orientationally into pShuttle2.The plasmid of pShuttle2-MAP4 was double-digested with restriction endonucleases NheⅠ and NocⅠ,and inserted BD Adeno-X~(TM) Virul DNA,named pAd2-MAP4.The non-recombinant adenovirus was screened out with PacⅠ, pAd2-MAP4 was linerized with SwaⅠ,and the recombinant virus genome was transfected into HEK293 cell line for packaging and amplification of Ad-MAP4 virus.The recombinant adenovirus was transfected into rat cardiac myocytes and MAP4 was identified by immunohistochemistry.Results The recombinant adenovirus-MAP4 was constructed successfully and the titer was about 2.3?10~(8) pfu/ml.The expression of MAP4 was enhanced at 48 h after the transfection.Conclusion We have successfully constructed a recombinant adenovirus Ad-MAP4 that has enforced the expression of MAP4 in vivo.

With the advances of genome-wide sequencing technologies and bioinformatics approaches, a large number of datasets of normal and malignant erythropoiesis have been gener-ated and made public to researchers around the world. Collection and integration of these datasets greatly facilitate basic research and clinical diagnosis and treatment of blood disorders. Here we provide a brief introduction of the most popular omics data resources of normal and malignant hematopoiesis, including some integrated web tools, to help users get better equipped to perform common analyses. We hope this review will promote the awareness and facilitate the usage of public database resources in the hematology research.

BACKGROUND: It is difficult to achieve satisfactory results with the traditional treatment of large-area skin defects and deep burns. OBJECTIVE: To test the treatment effect of an active dressing film made of a mixture of fibrin glue and bone marrow mesenchymal stem cells (BMSCs) for repairing burn wounds on the skin of rats. METHODS: Two scald wounds were made on the back of each rat. A total of 30 scald wounds were randomly divided into 3 groups, with 10 wounds in each group. In the experimental treatment group, the scald wounds were covered with the fibrin glue and BMSC mixture. The wounds of the experimental control group were covered with fibrin glue only. No intervention was administered to the blank control group. Thirty days after treatment, pathological sections were cut from the scalded local tissues of all rats from the 3 groups and observed with a microscope. RESULTS: The speed of scald wound healing in the experimental treatment group was faster than the other 2 groups. In the experimental treatment group, histopathological analysis revealed that the sebaceous glands showed obviously proliferous at the edge of the new tissue and gradually extended to the deep dermal layer of the new tissue. CONCLUSION: BMSCs may have an active role in promoting skin tissue repair and generating skin appendages. Allogeneic BMSCs mixed with fibrin glue can contribute to the quick formation of a film-like gel over the scald wounds, which might be of significance for emergency treatment and skin-grafting operations.
Animals ; Bandages ; Bone Marrow* ; Burns ; Emergency Treatment ; Fibrin Tissue Adhesive* ; Mesenchymal Stromal Cells* ; Rats* ; Sebaceous Glands ; Skin* ; Skin, Artificial ; Tissue Engineering ; Wound Healing ; Wounds and Injuries