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Background Application of confocal microscopy in the investigation of ocular surface system in living eye have been greatly extended in two decades.In vivo confocal microscopy allows the evaluation of the normal or pathological tissue at the cellular level.However,seldom study about the morphology of normal in vivo human bulbar conjunetiva under the confocal microscopy was induced. Objective Present study was to analyze the appearance of normal bulbar conjunetiva cells under the in vivo confocal microscopy. Methods Laser scanning confocal microscopy was used to examine the morphological characteristics of the bulbar conjunetiva in 21 eyes of 15 healthy volunteers.The parameters of confocal microscopy were as follows:resolution 1 μm,wave length 670 nm,field range 400 μm×400 μm.The epithelial cell numbers,dendritics cells density and goblet cells density from superior,inferonasal,nasal and temporal bulbar conjunctiva were calculated respectively and compared and imaged by the HRT3 Rostock Cornea Modual. Results The superficial epithelial cells of the bulbar conjunctiva was seen with the small cell nuclei and blur border.The borders of basal epithelial celt were clearly visible without cell nuclei.The presumed goblet cell presented with a large hyperreflective oval-shaped cell with relatively homogeneous brightness,crowed in groups or mainly dispersed.The orfices at the epithelial surface represented the goblet cells,showing some open and expel contents.The dendritic cell appeared to be hyperreflective corpuscular particles with visible processes among conjunetival epithelial cells.A few dense white fiber meshwork was exhibited in conjunctival stroma with the traverse blood vessels containing cellular elements.The superficial and basal epithelial cell densities were 2556±692and 2985±376 cells/mm2 respectively,and overall goblet and mature dendritic cells densities were 77±39 and 26±35 cells/mm2 respectively.Significant differences were proved in globet cell density and dendritic density among different conjunetival zones(P=0.001,P=0. 000),however,the alteration of conjunetival epithelium cells was insignificant in different area(P=0.204,P=0.130).Conclusion Confocal microscopy is a useful tool for the study of morphology of human bulbar conjunctiva cells in vivo.which offer a valuable aid in the diagnosis of ocular surface disease.
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Chylothorax is an uncommon disease where fatty fluid accumulates within the chest cavity. Conservative management, including repeated thoracentesis or pleurodesis, seems to be suitable to most cases. Herein, we present a case of efficacious pleurodesis by intrapleural injection of Sapylin, a streptococcus preparation, for the treatment of chylothorax. A 52-year-old non-smoking female farmer was diagnosed as idiopathic chylothorax after we ruled out possible causes including chest trauma, lymphoma, lung cancer, filariasis, tuberculosis, and etc. Two-time intra-thoracic injection of 3 Klinische Einheit (KE) Sapylin achieved rapid and effective control of chylothorax with no severe side effects. Sapylin may facilitate pleurodesis by producing a strong inflammatory response.
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Feminino , Humanos , Pessoa de Meia-Idade , Vacinas Bacterianas , Usos Terapêuticos , Produtos Biológicos , Usos Terapêuticos , Quilotórax , Tratamento Farmacológico , Patologia , Streptococcus , Química , Tomografia Computadorizada por Raios XRESUMO
<p><b>OBJECTIVE</b>To identify a new peptide deformylase (PDF) gene (Genebank Accession AY238515) from Enterococcus faecium and to establish a new screening model targeted on PDF.</p><p><b>METHODS</b>A new PDF gene was identified by BLAST analysis and PCR and was subsequently over-expressed in the prokaryotic expression host E. coli B121(DE3). Over-expressed protein was purified for enzymatic assay by metal affinity chromatography and a new screening model was established for novel antibiotics.</p><p><b>RESULT</b>A new PDF gene of Enterococcus faecium was identified successfully. Ten positive samples were picked up from 8000 compound library and the microbial fermentation broth samples.</p><p><b>CONCLUSION</b>A new PDF of gene Enterococcus faecium was first identified and the model had a high efficacy. Positive samples screened may be antibacterial agents of broad spectrum.</p>
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Amidoidrolases , Genética , Sequência de Aminoácidos , Antibacterianos , Usos Terapêuticos , Clonagem Molecular , Avaliação Pré-Clínica de Medicamentos , Enterococcus faecium , Genética , Fluorescamina , Fluorescência , Concentração de Íons de Hidrogênio , Indicadores e Reagentes , Cinética , Modelos Moleculares , Dados de Sequência Molecular , TemperaturaRESUMO
<p><b>OBJECTIVE</b>To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs.</p><p><b>METHODS</b>To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E. coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain.</p><p><b>RESULTS</b>With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K.</p><p><b>CONCLUSIONS</b>The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.</p>
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Humanos , Bioensaio , Western Blotting , Catepsina K , Catepsinas , Genética , Linhagem Celular , Clonagem Molecular , DNA Complementar , Avaliação Pré-Clínica de Medicamentos , Escherichia coli , Genética , Expressão Gênica , Osteoporose , Tratamento Farmacológico , Papaína , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
At the beginning of the 1980s, a concept of viable but non-culturable(VBNC) was suggested. VBNC is a survival strategy adopted by microorganisms when they are exposed to environmental stress. This article try to make a summary of research of the conditions of VBNC formation, recovery of culturability and methods of VBNC cells detection. In addition, introduces the first growth factor of microorganisms-Rpf.
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This project is targeted on exploring some improving approaches to isolate and culture the microorganisms which are difficult to be isolated and cultured through the conventional ways. The results showed that betaine, sodium pyruvate, SOD and catalase are helpful for increasing the total number and variety of isolated strains. A kind of combined method was also used to isolate the micro-colony which can not be seen by naked eyes on the plates. Totally 52 Actinomycetes and 103 bacteria and 17 fungi were obtained from 4 soil samples using the above methods. 4. 325% microorganisms were obtained as positive strains to inhibit the growth of some kinds of test bacteria, which is higher than the percent using generally isolated ones. These microbial natural products may remain an important resource for the drug discovery.
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Streptomyces sp.X-435 isolated from a soil sample collected in the suburbs of Beijing was proved to be a produce Virginiamycin.To improve the productivity of the Virginiamycin of Streptomyces sp.X-435,the spores of strain X-435 were treated with UV.The three types of colony,strawhat,wrinkled,blad,were isolated on Gaose's medium plates after mutation.Among them,the colonies of strawhat type exhibited positive mutation and were picked up as objects of screening.After five generation of mutation,the mutant F5-25-u-28 was selected which potency of Virginiamycin was about 20times higher than that of the beginning strain by flask fermentation and was also genetic stable.