Objective To prepare a new type of micropore porcine acellular dermis matrix with the aid of laser (LPADM),and to validate the healing effect of the LPADM through the phrase Ⅰ composite transplanting on the back of the full- thickness skin defects in SD rats.Methods In vitro,the allogeneic fibroblasts were separately cultured with the LPADM (LPADM group) or the non-pore PADM (non-pore LPADM group),while fibroblasts cultured by pure medium were used as control.After culture of 1 day,3 days and 5 days,the contents of IL-10,IL-6,TGF-β1,LN,VEGF expressed by fibroblasts were determined by double-antibody sandwich ELISA method.In vivo,the phrase Ⅰ transplantations of LPADM graft with split-thickness autologous skin were carried on the backs of the full-thickness cutaneous defects of SD rats.The healing condition was observed and analyzed by histological tests.Results The differences of the absorbance value between the LPADMgroup,PADM group and control group in each day were not statistically significant (F=0.050-1.763,P＞0.05).The transplantation of LPADM graft with split-thickness autologous skin graft resulted in high rate of surviving without signs of rejection 3 weeks later.After 1-month of transplantation,the regenerated skin was well enough to be lifted without any serious scars.Conclusions The phrase Ⅰ transplantation of LPADM graft with split-thickness autologous skin graft can accelerate the healing process of full-thickness skin wounds with high biological safety.

Objective:To analyze the volatile components in compound Shenqiyizhi powder. Methods: The technique of head-space solid phase micro extraction( HS-SPME) with gas chromatography-mass spectrometry ( GC-MS) was applied to study the vola-tile components in the compound powder. The compound essential oil was analyzed by steam distillation( SD) with GC-MS. Results:A total of 117 kinds of volatile components were detected, and among them, 70 kinds of chemical compositions were identified from the compound powder, accounting for 94. 46% of the total volatile components. The component with the highest content wasβ-himachalene followed by ylangene (15. 56%) and β-asarone (7. 36%). Totally 66 kinds of volatile components were detected from the compound essential oil by SD-GC-MS, and among them, 43 kinds were identified, accounting for 77. 41% of the total volatile components. The results indicated that the main volatile components were β-asarone (16. 73%), eugenol methyl ether (6. 74%) and ylangene (5. 81%). Conclusion:The main volatile components in Shenqiyizhi powders areβ-cedrene, ylangene,β-asarone,α-asarone and el-emicin etc. The research can provide scientific basis for the analysis of volatile components.

TNF-related apoptosis-inducing ligand (TRAIL) is a member of factor TNF family, which could be potentially developed as novel antitumor agent due to its selective and efficient induction of apoptosis in tumor cells. Gene recombinant expression is an important tool for production of pharmaceutical protein. In this paper, the gene encoding human soluble TRAIL (114-281aa fragment) was cloned by PCR and then inserted into the Pichia Pastoris expression vector pPIC9K. The transformants were double-screened on plates containing neomycin G418 and many clones with high levels of G418-resistance were selected for further studies on protein expression. The recombinant human soluble TRAIL was secreted into the BMMY media under the condition of 3% methanol. And the recombinant protein was purified to homogeneity (-80% purity) by using Ni-agarose affinity chromatography. The yield of this protein is about 1-2 mg per liter culture. Cell viability assays demonstrated that human soluble TRAIL was cytotoxic in both leukemia cells Jurkat and lung cancer cells A549. After treatment with 0.05 microg/ml TRAIL, the survival rate of Jurkat cells was about 10%. The expressed TRAIL showed dose-dependent cytotoxicity in A549 cells within the range of 0.1-1 microg/ml. When the protein concentration reached 1 microg/ml, the survival rates of A549 cells were about 30%. However, the recombinant human soluble TRAIL did not show obvious cytotoxicity in human skin fibroblast cells (HSF) at concentrations tested. There results demonstrate that human soluble TRAIL is selectively cytotoxic in tumor cells. The expression system constructed in this experiment might contribute to further production of soluble TRAIL and TRAIL-based novel fusion proteins in large quantities.
Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cloning, Molecular ; Cytotoxicity Tests, Immunologic ; methods ; Genetic Vectors ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; genetics

Objective To understand the status of knowledge and attitude of schistosomiasis control among the human populafion in schistosomiasis transmission-controlled area of Changqiu mountainous areas.Methods The subjects were selected by the random cluster sampling method among residents and students in these alias,then they were investigated by questionnaire.Resuits A total of 150 residents and 209 students were selected.There were 60% of the residents whose awareness rates of the knowledge on schistosomiasis control were above 90%.The correction rates of the questions in residents were between 99.30% and 100%.and the awareness rates about the questionswhether re-infection would occure after schistosomiasis was cured and the remedy for schistosomiasisof female adults were both 75.40%.The correction rates of the two question8 on attitude and behaviour of schistosomiasis control in adults were above 80.00%.In students'questionnaires.the awareness rates of knowledge on schistosomiasis control were above 90%.except the two questions on the shape of the snailand the infection-risk months of schistosomiasis.The correction rates of attitude and behaviour of schistosomiasis control were aslo above 80%.Conclusion The correction rates of knowledge,attitude and behaviour of schistosomiasis control of adults and students have reached the national goal of schistosomiasis control in 2008.

Adult ; Female ; Heat-Shock Proteins ; genetics ; Humans ; Leprosy ; diagnosis ; genetics ; microbiology ; Male ; Middle Aged ; Mycobacterium leprae ; pathogenicity ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Skin ; metabolism

OBJECTIVETo define the main genotypes in Guizhou agricultural areas by molecular epidemiologic investigation of 21 Borrelia burgdorferi sensu lato of Lyme disease spirochetes and to provide the scientific bases for formulating a preventive policy.

METHODSPolymerase chain reaction (PCR) technique was used to amplify the 23S(rrl)-5S(rrf) intergenic spacer, and amplified products were analyzed by restriction fragment length polymorphism (RFLP) and nucleotide sequencing.

RESULTSThere were two genospecies in the strains: 20 strains belong to Borrelia valaisiana, 1 strain is Borelia sp.

CONCLUSIONBorrelia valaisiana was the main genotype in Guizhou agricultural areas. The harmness of B. valaisiana to human being has been confirmed. In order to efficiently prevent the harmness of agent to the people in Guizhou agriculture areas, we should study the risk further.

Base Sequence ; Borrelia burgdorferi ; classification ; genetics ; China ; DNA, Bacterial ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Humans ; Lyme Disease ; microbiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 23S ; genetics ; RNA, Ribosomal, 5S ; genetics ; Ribotyping ; Sequence Analysis, DNA

BACKGROUNDAntimicrobial peptides, including cathelicidin LL-37, human beta defensin (HBD)-2, and HBD-3, are important elements of the innate immune response and involved in modulation of the adaptive immunity, and they also play an important role in cutaneous defense against Mycobacterium tuberculosis.

METHODSThe fresh skin tissues and paraffin-embedded biopsy samples from three cutaneous tuberculosis, two tuberculids, and ten healthy individuals were collected. The expressions of LL-37, HBD-2, and HBD-3 mRNA in the lesions of three cutaneous tuberculosis and two tuberculids were detected by quantitative real-time polymerase chain reaction; the protein expressions were detected by immunohistochemistry and Western blotting methods.

RESULTSThe expressions of LL-37 mRNA and protein in the lesions of cutaneous tuberculosis and tuberculids were similar to that of normal skin. The expression of HBD-2 mRNA had an increasing trend in the lesions of cutaneous tuberculosis and tuberculids compared with that of normal skin; however, the expression of HBD-2 protein in the lesions of cutaneous tuberculosis had a decreasing trend compared with that of normal skin, and the expression of HBD-2 protein in the lesions of tuberculids was similar to that of normal skin. The expressions of HBD-3 mRNA and protein in lesions of cutaneous tuberculosis and tuberculids were similar to that of normal skin.

CONCLUSIONSOur study indicated that the expression of HBD-2 and HBD-3 mRNA and protein in lesions of cutaneous tuberculosis may be not consistent with that of tuberculids. However, an inherent limitation of the present study was that the sample size was small, and the roles and regulation mechanisms of LL-37, HBD-2, and HBD-3 in cutaneous tuberculosis and tuberculids need to be further investigated.

Adult ; Aged ; Antimicrobial Cationic Peptides ; genetics ; Female ; Humans ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Tuberculosis, Cutaneous ; metabolism ; beta-Defensins ; genetics

Cytochrome C plays important roles in electron transferring, oxidative stress and apoptosis. In this study, soluble cytochrome C was accumulated in cytoplasm of E. coli by utilizing the co-expression of human cytochrome c and yeast heme lyase from a single plasmid. After ultrasonic disruption of the bacteria, a lot of contaminated proteins were discarded by addition of 350 g/L ammonium sulfate into the supernatant. Then the recombinant human cytochrome C was purified to 80% homogeneity after two times cation exchange chromatography on SP-Sepharose Fast Flow. Yields of cytochrome C greater than 10 mg per liter culture were attained. This efficient system for producing human cytochrome C is helpful for us to understand the roles of this protein in biological processes and therapy of human diseases relevant to apoptosis and oxidative stress.
Cytochromes c ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

Due to their lower risk for induction of resistance, membrane-active antimicrobial peptides with anticancer effect are attractive in cancer therapy. Because cell binding contributes to the cytotoxicity of peptide, it is possible to enhance the cytotoxicity of antimicrobial peptide in tumor cells by conjugation to a cell-penetrating peptide (CPP). In this paper, a fusion peptide MPGA by conjugation of antimicrobial peptide MP to CPP Antp at its N-terminus was constructed. After compared the cytotoxicity of unconjugated MP with that of the fusion peptide, it was found that MPGA showed higher cytotoxicity than that of unconjugated MP. And the fusion peptide MPGA induced cell death in tumor cells by membrane disruption. These results demonstrated that the cytotoxicity of antimicrobial peptide can be significantly enhanced by conjugation to CPP, which might be an effective way to develop novel anticancer drugs.
Antimicrobial Cationic Peptides ; pharmacokinetics ; pharmacology ; Antineoplastic Agents ; pharmacokinetics ; pharmacology ; Cell Line, Tumor ; Cell Membrane Permeability ; drug effects ; Humans

OBJECTIVETo compare the effects of small fenestra stapedotomy with semiconductor diode laser and microdrill in patients with otosclerosis.

METHODSTwenty-six patients (29 ears) undergoing stapedotomy with semiconductor diode laser and 19 patients (21 ears) with microdrill were compared for the hearing results and complication rates.

RESULTSNo statistically significant differences were found in postoperative speech frequency and high frequency pure tone average in closing the air-bone gap between the two groups. The ears treated by stapedotomy with semiconductor diode laser showed significantly better preoperative minus the postoperative air-bone gap and milder dizziness.

CONCLUSIONIn spite of the good hearing outcomes in both groups, small fenestra stapedotomy with semiconductor diodelaser can achieve better results and reduce the incidence of complications.

Adult ; Aged ; Female ; Fenestration, Labyrinth ; methods ; Hearing Tests ; Humans ; Lasers, Semiconductor ; Male ; Middle Aged ; Otosclerosis ; physiopathology ; surgery ; Stapes Surgery ; instrumentation ; methods ; Young Adult