Practice and discussion of cultural integration in multi-campus hospitals / 中华医院管理杂志

The multi-campus mode is an important way to give full play to the advantages of public hospitals and promote the expansion of high-quality medical resources and balanced regional layout. The authors summarized the practical experience of the Second Affiliated Hospital Zhejiang University School of Medicine in promoting multi-campus cultural integration, including vertical dimensional initiatives including raising cultural construction to a strategic level, improving the working mechanism of cultural construction, and building a distinctive cultural identity system; horizontal dimensional initiatives including creating equal status and intergroup cooperation conducive to cultural integration, building a variety of forms of the main cultural communication platform, and building a unified and diverse cross-campus communication bridge. Through cultural integration, the internal cohesion of the hospital was enhanced and the influence of the hospital brand was improved. The authors suggested that cultural integration should always be based on the principle of " seeking common ground while preserving minor differences" , focusing on the construction of systems and standards, and focusing on the construction of communication platforms.

Effect of passage number on the activity and spontaneous reverse mutation of Ames test strains / 上海预防医学

Objective We explored the stability of the bacteria strains used in the Ames test to provide a basis for determining the appropriate passage number at which the biological characteristics of the strains would not change. Methods The Salmonella typhimurium (TA97a, TA98, TA100 and TA102 strains) were selected as the experimental strains.The original frozen strains and frozen strains with different passage times were used to compare the biological characteristics and the spontaneously reverting colonies. Results The biological characteristics of four kinds of strains, which were histidine deficiency, lipidpolysaccharide barrier defect, ampicillin resistance, UV sensitivity, and tetracycline resistance, did not change at F1-F6 generation when compared with the F0 generation.However, as for the number of spontaneously reverting colonies, a statistically significant difference (P < 0.05) occurred at F3 generation when compared with F0 generation for the TA97a strain, and a significant difference (P < 0.05) occurred at F4 generation for TA100 and TA102 strains. Conclusion Passage number of strains used in Ames test could affect their spontaneous reversion mutation rate.The passage number should be less than 4 for TA98、TA100、TA102 strains, and less than 3 for TA97a in Ames test.

Impact of community subculture on high-risk sexual behavior of men who have sex with men based on health belief model / 中南大学学报(医学版)

OBJECTIVES@#To explore the Impact of community subculture of men who have sex with men (MSM) on the occurrence of high-risk sexual behavior based on the health belief model.@*METHODS@#A qualitative research method was used to conduct in-depth interviews with 17 MSM by one-to-one and half-structured way, and thematic analysis was used to analyze the data.@*RESULTS@#There were several factors for high-risk sexual behavior in MSM subculture, such as trust, subjective assessment for partner or personal health status, the role in inserting, awareness of HIV infection among partners, perception of HIV and homosexual discrimination, difficulty in maintaining a fixed partner, family responsibility,and so on. Self-efficacy also affected MSM's high-risk sexual behavior.@*CONCLUSIONS@#High-risk sexual behavior in MSM population is influenced by individual, group, and intra-circle subculture. Cognitive bias for HIV infection in MSMs can be intervened by constructing a preventive intervention model for high-risk sexual behavior.

Enseñanza contextualizada para miembros de la subcultura del heavy metal a través de técnicas teatrales / Contextualized Teaching and Learning using Drama Techniques for Members of the heavy metal Subculture / Ensino Contextualizado para Membros da Subcultura do heavy metal através de Técnicas Teatrais

Este artículo da cuenta de la investigación cualitativa que pretende incluir a la subcultura del metal en el aula de inglés como lengua extranjera (ILE) en Chile, utilizando la enseñanza contextualizada y las técnicas teatrales. En primer lugar, se otorgan algunos antecedentes relacionados con el ILE en Chile. En segundo lugar, se desarrolla una revisión bibliográfica, en la que se describe a las subculturas juveniles, los metaleros como subcultura, la motivación para aprender ILE, la enseñanza contextualizada y finalmente las técnicas dramáticas. En tercer lugar se entrega la metodología de investigación utilizada (entrevistas y observación no participante) y algunos de los resultados obtenidos. En último lugar, se presentan conclusiones y una propuesta pedagógica que se sirve de esta investigación para incluir la subcultura juvenil del metal en el aula de ILE.

This article describes qualitative research on the inclusion of the heavy metal youth subculture in English as a Foreign Language (EFL) classrooms in Chile, using contextualized teaching and learning as well a drama techniques. Background information related to EFL teaching in Chile is provided followed by a literature review that describes heavy metal as a youth subculture, motivation for learning EFL, contextualized teaching and learning and finally drama techniques. The article goes on to detail the research methodology used (interviews and non-participant observation) and the results of the study are highlighted. The authors present their conclusions and suggest a pedagogical proposal that brings together all of the concepts examined by this research for the incorporation of heavy metal subculture in EFL lessons.

Este resumo descritivo relata a pesquisa qualitativa que visa incluir a subcultura do heavy metal em aulas de Inglês como Língua Estrangeira (ILE) no Chile, usando o ensino contextual e técnicas teatrais. Em primeiro lugar, alguns antecedentes relacionados com à ILE no Chile são concedidos. Em segundo lugar, desenvolve-se uma revisão bibliográfica na qual são descritas as subculturas juvenis, os metaleiros como subcultura, a motivação para aprender ILE, o ensino contextualizado e finalmente técnicas dramáticas. Em terceiro lugar, apresenta-se a metodologia de pesquisa utilizada (entrevistas e observação não participante) e alguns dos resultados obtidos. Finalmente, apresentam-se conclusões e uma proposta pedagógica que utiliza essa pesquisa para incluir a subcultura jovem do heavy metal na aula de Inglês como Língua Estrangeira (ILE).

Subculture, cryopreservation and recovery of osteoclasts / 中国骨伤

<p><b>OBJECTIVE</b>To explore the feasibility of passage, cryopreservation, and recovery of osteoclasts in order to develop new techniques facilitating osteoclast research.</p><p><b>METHODS</b>Passage of osteoclasts: adult male SD rat(SPF grade, weight of 250 g) was sacrificed and the abdominal aorta was exposed for blood draw. Monocytes isolated from peripheral circulation was treated with RANKL and M-CSF for 2 weeks. After formation of osteoclasts, they were trypsinized with pipetting, centrifuged, re-suspended with α-MEM containing RANKL and M-CSF, and cultured in 6 well-plates and 35 mm culture dishes. Freezing of osteoclasts: trypsinized osteoclasts were centrifuged and resuspended with DMSO, FBS, α-MEM (1:2:7), and were stored in liquid nitrogen(-196 °C). Recovery of osteoclasts: frozen osteoclasts were taken out of liquid nitrogen tank and thawed quickly at 37 °C in water bath. After wash with PBS, the cells were resuspended with α-MEM containing RANKL and M-CSF, and were cultured in 6 well dishes and 35 mm culture dishes. Meanwhile, cells were checked with inverted phase contrast microscope and observed in the live cell station for real time imaging. TRAP staining was performed 3 days after plating.</p><p><b>RESULTS</b>Trypsinization together with pipetting and shaking can detach the adherent osteoclasts, and the resuspended cells can be used for passage and storage in liquid nitrogen. The passaged cells became fully attached to the culture dishes in 2 hours, and the multinucleated feature could be clearly seen. The osteoclasts recovered from liquid nitrogen could completely spread out for 2 to 3 hours so that the multinucleated cells were clearly seen. These cells were still TRAP positive.</p><p><b>CONCLUSIONS</b>Although osteoclasts strongly adhere to the bottom of culture dishes, a large majority of the osteoclasts can be detached after appropriate digestion with trypsin, pipetting and shaking. These cells can be used for passage and cryopreservation. After recovering from liquid nitrogen, these cells still preserve the viability and the feature of osteoclasts. The results provide a new and powerful tool for future study of osteoclast biology.</p>

David Matza: perspectiva criminológica de la deriva a la delincuencia juvenil / David Matza: criminological perspective of minors drifting towards juvenile delinquency / David Matza: perspectiva criminológica da deriva à delinquência juveni

El fenómeno de la delincuencia juvenil es un tema que aún en la actualidad sigue generando polémica; las líneas de investigación enfocan su esfuerzo en la búsqueda de las razones por las cuales un menor entra en el mundo delincuencial. Sin embargo, omiten la existencia de autores que dedicaron sus estudios a dar respuesta a tal cuestión. Un autor clave en el estudio del fenómeno delincuencial es el sociólogo estadounidense David Matza, pionero en las teorías de carácter social que pretenden dar respuestas a las incógnitas planteadas dentro de la temática juvenil; el presente artículo se enfocará en el análisis y reflexión de los principales postulados de su teoría de las subculturas delincuenciales, por medio de una perspectiva basada en la interacción, lo cual nos mostrará que es una subcultura a la par de la cultura convencional, con miembros que no distan de aquellos jóvenes que son considerados normales por la mayoría de la sociedad. Así se explica por qué algunos jóvenes se aproximan al quebrantamiento de la ley.

The phenomenon of juvenile delinquency has continued to be an ongoingly controversial issue; current research lines are focusing their efforts in search of the reasons why a minor ends up entering the delinquent world. Nevertheless, they fail to acknowledge the presence of authors having devoted their studies in trying to provide an answer to that question. A leading scholar in the study of this phenomenon is sociologist David Matza from the United States, a pioneer in theories of social nature attempting to give a solution to all the unknowns posed within the context of juvenile topics. This article is focused on analysis and reflection about the major premises of his theory dealing with delinquent subcultures from a perspective based on interaction. This will show us, therefore, that this is a subculture in keeping with the conventional culture, with members not far from those young people considered normal by a majority of society. This is the explanation concerning why some of them are quite close to breaking the law.

O fenômeno da delinquência juvenil é um assunto que ainda no tempo atual continua a gerar polémica; as linhas da investigação focalizam seu esforço na busca das razões para que um menor entre no mundo delinquencial. Não obstante, omitem a existência dos autores que dedicaram seus estudos à resposta da questão. Um autor chave no estudo do fenômeno delinquencial é o sociólogo americano David Matza, pioneiro nas teorias do caráter social que tentam dar a respostas às incógnitas expostas dentro da temática juvenil; esse artigo focara-se na análise e na reflexão dos postulados principais de sua teoria das subculturas delinquenciais, por meio de uma perspectiva baseada na interação, que nos mostrará que é uma subcultura no mesmo nível da cultura convencional, com membros que não distam daqueles jovens que são considerados normais pela maioria da sociedade. Assim explica-se porquê alguns jovens aproximam-se a infringir a lei

Optimization of tissue culture conditions for Pfaffia paniculata / 中草药

Objective: To establish and optimize the technique of tissue culture and rapid propagation of Pfaffia paniculata. Methods: Using MS and 1/2MS basic media, the effects of various combinations of plant growth regulators (6-BA, NAA, IBA, and IAA) on the seedlings subculture and rooting culture were studied by orthogonal design. Results: The effective medium for cluster buds-inducing and subculture was MS + 6-BA 1.5 mg/L + NAA 0.1 mg/L + IBA 0.2 mg/L, and the propagation coefficient was over 6.0 per 30 d; The best rooting medium was 1/2 MS + NAA 1.0 mg/L + IBA 0.2 mg/L, and the rooting rate was 100%. The rooting seedlings were transplanted in well-drained sand bed and the survival rate was 98%. Conclusion: The method in the experiment could provide a basis on protecting the mild resources of P. paniculata exploring the artificial resources, and discussing the new way breedings.

In vitro Regeneration of Plantlets from Leaf and Nodal explants of Aristolochia indica L.- An Important Threatened Medicinal Plant

Objective: An efficient reproducible protocol has been developed for in vitro regeneration of plantlets from leaf and nodal explants of Aristolochia indica L. Methods: Wild grown plants Aristolochia indica L. were collected and grown in the departmental garden. Leaf and nodal segments (0.5-1.0 cm) from young healthy plants were first washed thoroughly under running tap water for 15 - 20 minutes and then treated with liquid detergent [5% (v/v) Tween-20] for 5-10 minutes. Later these explants were washed with double-distilled water for 5 minutes. Subsequently, explants were immersed in 70% (v/v) ethanol for 2 - 3 minutes and washed with sterile glass double distilled water for 2-3 times. Eventually, the explants were treated with an aqueous solution of 0.1% (w/v) HgCl2 for 1 - 2 minutes and rinsed for two-to-three times in sterile ddH2O to remove all traces of HgCl2. The sterilized explants were inoculated aseptically onto solid basal Murashige and Skoog’s medium with different concentrations and combinations of BAP and NAA for in vitro regeneration of plants. Results: Both leaf and nodal explants cultured on MS medium supplemented with 0.8 mg/L BAP developed into mass of callus. These calli were subcultured for the induction of shoots and roots. Shoots were induced from both calli on MS medium supplemented with 0.8 mg/L BAP+0.5 mg/L NAA. Roots were induced from in vitro shoots on MS medium supplemented with 0.8 mg/L NAA for 4 weeks. Nodal explants were more regenerative with 95 % response compared to leaf explants with 85%. Finally, these in vitro regenerated plantlets were hardened, acclimatised and successfully transferred to the field. Conclusions: The present protocol for in vitro regeneration of Aristolochia indica L. can be used to make this plant available throughout the year for traditional healers, pharmaceutical usages, germplasm conservation, commercial cultivation, and also for the production of secondary metabolites.

Indicadores estructurales y conglomerados de actores en la red social de una subcultura urbana / Indicadores estruturais e conglomerados de atores na rede social de uma subcultura urbana / Structural indicators and clusters of actors in the social network of an urban subculture

Este estudio buscó identificar la estructura de socialización de una subcultura urbana con la medición de indicadores estructurales de su red social e identificación de subagrupaciones. Se empleó el instrumento Arizona Social Support Interview Schedule (ISSIS) con 11 miembros de la subcultura roqueros. Se procesaron los datos mediante el análisis de redes sociales (ARS) con el software Ucinet, para detectar conjuntos de actores con el procedimiento de conglomerados jerárquicos; y los gráficos de la red se crearon con NetDraw. Se identificaron niveles moderados de indicadores estructurales y las agrupaciones presentaron un número amplio de actores según el criterio de atracción por similaridad. Los subgrupos en la red fueron escasos, lo que muestra a la subcultura como una unidad social cuya integración se da por vinculaciones estrechas entre sus miembros.

The objective of this study was to determine the socialization structure of a subculture or urban tribe, by measuring the structural indicators of its social network and identifying sub-groupings. Eleven members of the rock subculture were evaluated through the Arizona Social Support Interview Schedule (ASSIS). The data was processed using the social network analysis (SNA) with the software Ucinet, and networks graphs were created with NetDraw. It was possible to identify moderate levels of structural indicators in which groupings of large numbers of actors under the criterion of attraction by similarity was significant. Finally, there were few subgroups within the network, which indicates that the subculture is a social unit where integration is achieved by close ties among its members.

Este estudo buscou identificar a estrutura de socialização de uma subcultura urbana através da análise de indicadores estruturais de rede social e a identificação de sub-agrupações. Utilizou-se o instrumento Arizona Social Support Interview Schedule com 11 membros da subcultura roqueiros. Os dados foram processados mediante a análise de redes sociais com o software Ucinet para detectar conjuntos de atores com o procedimento de conglomerados hierárquicos, e os gráficos da rede foram criados com NetDraw. Identificaramse níveis moderados de indicadores estruturais e as agrupações presentaram um número amplo de atores segundo o critério de atração por similaridade. Os subgrupos na rede foram escassos, o que mostra a subcultura como uma unidade social cuja integração ocorre por vinculações próximas entre seus membros.

Optimal subculture methods for passaging and growing human epidermal keratinocytes.

Background: Human epidermal keratinocytes (HEKs) cover the outer layer of the skin and play a key role in wound repair. Although the methods for isolation and cultivation of primary HEKs from epidermis have been used successfully in both laboratory and clinical settings, the ability to subculture (passage) these cells has yet to be established and is the primary factor hindering their usage. Objectives: We conducted this study to identify optimal subculture conditions for HEKs. Methods: We first harvested the primary HEKs from prepuce tissue specimens, and then compared three different reagent compositions (0.25% trypsin, 0.25% trypsin plus 0.01% EDTA, and 0.25% dispase digestion solution) for various periods of time at 4oC with the conventional 0.25% trypsin or 0.25% trypsin plus 0.01% EDTA digestion at room temperature. Results: Our data indicated that the cold digestion conditions yielded higher cell numbers and more viable cells than the conventional methods. Furthermore, the subcultured HEKs also adhered and grew better after four hours of a 0.25% trypsin cold digestion or after six hours of a 0.25% dispase cold digestion. These procedures produced higher numbers of HEK passages than that commonly seen experienced with conventional methods. Conclusion: The data from the current study demonstrated that the optimal subculture condition for passaging and growing HEKs in vitro is four hours digestion with 0.25% trypsin.

Effect of adipose tissue processing procedures in culture result: a study preliminary.

Background: There are various methods of processing adipose tissue before culture, depending on the adipose tissue samples. The aim of this study is to compare several modifications of culturing and sub-culturing procedures of adipose tissue to fit the condition in our laboratory. Method: This is a descriptive study that was done in the Immunology and Endocrinology Integrated Laboratory, University of Indonesia, from October 2009 to April 2010. Three adipose tissue processing procedures, various amount of seeding and two subculture methods were compared in term of cell yield and time needed. In the first procedure, collagenase-1 digestion was done in 30minutes, cell seeding were 24,000 and 36,000 per flask; in the second procedure, collagenase-1 digestion was done in 60minutes, cell seeding were 24,000, 48,000, and 72,000 per flask; and in the third procedure, the adipose tissue remnants from the first procedure were again digested for another 45 minutes, cell seeding were 74,000, and 148,000 per flask. Difference in subculture methods were the presence or absence of washing step. Result: Procedure 1 yielded the lowest amount of cell, and after culture, the cells grew very slow, and was contaminated before harvest of primary culture. Procedure-2 and -3 succeeded to yield primary cultures. Some of the cultures were contaminated, so that further subculture was not applicable, and only one tissue processing procedure (procedure 2: 60 minute collagenase-1 digestion, without lysis buffer, cell seeding 48,000 and 72,000) could complete the three subcultures. Though some of the procedures could not be completed, final result could be concluded. Conclusion: In this preliminary study, 60 minute colagenase-1 digestion with intermittent shaking every 5 minutes and cell seeding around 50,000 or more, followed by subculture method without washing step gave the best result.

Heterothallic Type of Mating System for Cordyceps cardinalis

Cordyceps cardinalis successfully produced its fruiting bodies from multi-ascospore isolates. However, subcultures of multi-ascospore isolates could not produce fruiting bodies after few generations. Fruiting body production also differed from sector to sector of the same isolate. Single ascospore isolates were then co-inoculated in combinations of two to observe the fruiting characteristics. Combinations of certain isolates produced perithecial stromata formation, whereas other combinations did not produce any fruiting bodies. These results show that C. cardinalis is a heterothallic fungus, requiring two isolates of opposite mating types for fruiting body production. It was also shown that single ascospore isolates are hermaphrodites.

Adequate Serial Monolayer Passage Number of Human Intervertebral Disc Cells for Cell Therapy: Growth and Phenotype of Cells

STUDY DESIGN: This is an in-vitro experimental study. OBJECTIVES: We wanted to analyze the changes in the growth and phenotype of human degenerative intervertebral disc cells depending on the frequency of subculture in an in vitro monolayer culture system. SUMMARY OF THE LITERATURE REVIEW: A subculture of disc cells is needed to obtain an adequate amount of disc cells for cell therapy, tissue engineering and analysis of the biological characteristics of degenerative disc cells. MATERIALS AND METHODS: The obtained intervertebral discs were divided into the nucleus pulposus (NP) and the annulus fibrosus (AF). The AF and NP cells were cultured in a monolayer manner, respectively. At each subculture time, we analyzed the morphological changes, the adhesion rate, the proliferation rate and the viability. The expressions of types I and II collagen and proteoglycan were analyzed at the mRNA gene level. RESULTS: Both the AF and NP cells gradually showed a fibroblast-like spindle shape while undergoing subculture. The adhesion rate was higher at the second and third times of subculture. The cell proliferation was the highest at the second subculture time. The viability was markedly lower prior to the subculture. On RT-PCR, the type II collagen expression was gradually decreased in the NP cells. In the AF cells, Type II collagen was not expressed from the second time of subculture. The expression of proteoglycan was gradually decreased in both. CONCLUSIONS: Following the 3rd subculture, the degenerative disc cells had completely changed their original growth and phenotypic characteristics. Therefore, we believe that it is not desirable for us to do passage cultures more than three times for cell therapy.

Subcultura no uso da força policial: uma análise do uso da força policial a partir da teoria das representações sociais / Subculture in the use of police force: an analysis of the use of police force following the theory of social representations

O presente artigo, inserido nas temáticas trabalhadas pela Psicologia Social e do Trabalho, busca analisar, por meio da Teoria das Representações Sociais, a presença de uma subcultura interferindo no uso da força policial, geralmente materializada na formação de grupos de policiais que utilizam esse instrumento. Tais grupos defendem que, se o policial não se impõe sempre pela força física durante uma intervenção policial, ele não consegue o respeito do cidadão abordado. A atuação das corporações policiais merece destaque nos diversos campos de estudos, pois diz respeito tanto aos indivíduos representantes das corporações, ou seja, os policiais, como à população beneficiária dos serviços prestados por essas instituições. Assim, este estudo de caso de caráter descritivo-analítico, fruto da conclusão de curso de graduação, tem como objetivo buscar uma melhor compreensão sobre a presença de preceitos subculturais no uso da força policial. Foi feito junto a uma amostra de policiais recém-formados e em processo de formação em um Centro de Ensino da Corporação da Polícia Militar do Estado de Minas Gerais. Dessa forma, partindo de análises de falas de companheiros de farda e da análise de informações obtidas com questionário, observou-se que, nas práticas policiais operacionais, fatores subculturais influenciam a conduta de determinados policiais, culminando na incorreta aplicação do uso da força. Buscou-se identificar, ainda, como esse fenômeno surge, suas principais características e as explicações teóricas possíveis para os resultados encontrados.

This article, inserted in the themes discussed in labor and social psychology, seeks to analyze, by means of the Theory of the Social Representations, the presence of a subculture interfering in the use of the police force, generally materialized in the formation of groups of policemen who make use of this instrument. Such groups argue that, if the policeman does not always impose himself through the use of physical strength during a police intervention, he is not able to get the respect of the approached citizen. The performance of police corporations deserves to be highlighted in the various fields of studies because they not only concern the individuals that represent the corporations, that is, the policemen, but also the population that benefits from the services provided by these institutions. Therefore, this case study of descriptive and analytical nature, resulting from an undergraduate‟s final coursework, has the goal of searching for a better understanding of the presence of subculture precepts in the use of police force. It was carried out with a sample of newly trained policemen and policemen in training process at a Center for Education of the Corporation of the Military Police of the State of Minas Gerais, Brazil. Thereby, starting out with analyses of the speeches of companions from the corporation, and from analyses of information obtained through questionnaire, it was observed that, in the operational police practices, subculture elements influence the conduct of certain policemen, culminating in the incorrect application of the use of the force. Moreover, this research tried to identify how this phenomenon appears, its main characteristics and the possible theoretical explanations for the obtained results.

The Biological Characteristics of Different Generation of Osteoblasts / 浙江中医药大学学报

[Objective]To investigate the discrepancy on biological characteristics of different subculture osteoblasts in vitro.[Methods] Compare the morphology,AKP activities and mineralization nodules of 1,3,5,7 and 9 subculture osteoblasts,and make statistic of the result.[Results] There was no discrepancy within 1,3 and 5 subculture osteoblasts,but the 7,9 subculture was significantly worse than subculture 1 in the morphology,AKP activities and mineralization nodules.[Conclusion] The osteoblasts within 5 subculture were suitable for experiment in vitro.

Osteogenic Potential of Human Mesenchymal Stem Cells During Serial Subculture / 대한정형외과연구학회지

PURPOSE: The purpose of this study was to evaluate the osteogenic differentiation potential of human mesenchymal stem cells during serial subculture. MATERIALS AND METHODS: Human bone marrow-derived MSCs were serially subcultured and then maintained in basal or osteogenic medium for 14 days. Then we performed FAC analysis, RT-PCR, alkaline phosphatase activity and stains. RESULTS: Human MSCs had different morphologies, immunophenotypes, and growth rates that were correlated with the length of serial subculture. The phenotype changed from small spindle-shaped cells at passage 1 into large cuboidal or flattened cells at passage 7. The osteogenic capacity of human MSCs decreased during serial subculture. Using RT-PCR, the mRNA levels of bone-specific genes, such as cbfa1/runx2 and osteocal-cin, decreased with increasing passage number. Strong positive staining was observed for ALP and Alizarin reds in osteogenic medium on day 14, but declined significantly with increasing passage number. CONCLUSION: We have shown that osteogenic potential of human MSCs decreased during serial subculture. This result can provide the helpful information to decide the timing of human MSC transplantation during in vitro culture expansion for treatment of bone defects and so on.

Cultura, complejo cultural y subcultura / Culture, cultural complex and subculture

"Cultura" es una noción utilizada por diversas disciplinas. Designa el orden humano como un todo, en aquello que es específico y lo diferencia del orden animal. "Complejo cultural" es una noción antropológica que se usa para designar una organización cultural determinada.

"Culture" is a notion used by various disciplines. It designates the human order as a whole, in that which is specific and differentiates it from the animal order. "Cultural complex" is an anthropological notion that is used to designate a specific cultural organization.

Secretory Differentiation of Serially Passaged Normal Human Middle Ear Epithelial(NHMEE) Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The purpose of this study was to subculture normal human middle ear epithelial (NHMEE) cells, investigate whether the subcultured NHMEE cells could have ability to differentiate into secretory cells, and establish a method to get cultured NHMEE cells for further study of human middle ear epithelial differentiation and secretion. MATERIALS AND METHOD: Freshly isolated epithelial cells from healthy middle ear mucosa were subcultured repeatedly after enzymatic disaggregation in serum-free medium on plastic tissue culture dishes. The subcultured cells were counted after every passage and tested for secretory differentiation in air-liquid interface (ALI) cultures. The apical secretion of cultured NHMEE cells were characterized by immunoblotting and Western blotting. RESULTS: Attachment rate of subcultured NHMEE cells was over 70% through every passage. Cells proliferated by 22 fold from passage-1 to passage-2 (P-2), but passage-4 cells did not proliferate. P-2 NHMEE cells in ALI cultures was stained with mucin antibody (H6C5) but not b-tubulin antibody. Cultured NHMEE cells secreted mucin and lysozyme. CONCLUSION: P-2 NHMEE cell cultures retained many important features of normal epithelium and were suitable for conducting many studies of human middle ear epithelial cell biology including cell differentiation and secretion.

Serially Passaged Normal Human Nasal Epithelial Cells: Morphology and Mucous Secretory Differentiation

BACKGROUND AND OBJECTIVES: The purpose of this study was to develop a subculturing technique that allows the formation of large amounts of normal human nasal epithelial (NHNE) cells without compromising the cells' ability to differentiate into secretory and ciliated cells. MATERIALS AND METHODS: Freshly isolated nasal epithelial cells, collected from normal inferior turbinates, were subcultured repeatedly in a serum-free medium on plastic culture dishes. The subcultured cells were tested for growth curve and electromicroscopic characteristics in air-liquid interface (ALI) cultures and for mucous secretory differentiation. RESULTS: The cultures grew rapidly during the first three passages, demonstrating a 20- to 40-fold expansion with each subculture. Ciliogenesis usually started on day 12 and the cultured cells formed cellular sheets exhibiting microvilli in the apical membrane, intracytoplasmic secretory granules, electron lucent, scant endoplasmic reticulum and complex tight junctions on the basolateral side. Passage-1 (P-1) and passage-2 (P-2) cells maintained their potential to differentiate into mucin-secretory and ciliated epithelial cells, and this potential was confirmed by immunocytochemistry with H6C5 and transmission electron microscopy. Although the number of NHNE cells on day 16 of culture decreased as the passage progressed from P-1 to P-3, the relative number of secretory cells did not significantly change. CONCLUSION: In conclusion, P-2 NHNE cell cultures retain the features of normal epithelium and are suitable for conducting many studies on upper airway cell biology.

Secretory Differentiation of Serially Passaged Normal Human Nasal Epithelial (NHNE) Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The purpose of this study was to subculture normal human nasal epithelial (NHNE)cells without compromising their ability to differentiate into secretory and ciliated cells and to study the effect of retinoic acid (RA)on mucous and serous secretions in each passaged cells. MATERIALS AND METHOD: Freshly isolated nasal epithelial cells from normal inferior turbinates were subcultured repeatedly in serum-free medium on plastic tissue culture dishes. The subcultured cells were tested after every passage for secretory differentiation in air-liquid interface (ALI) cultures. The apical secretion of cultured NHNE cells was characterized by immunoblotting and Western blotting. RESULTS: Cultured NHNE cells secreted mucin and lysozyme. RA was essential for mucociliary and secretory differentiation. The epithelium became squamous and mucin secretion decreased when RA was deleted from the culture media. Cells from passage 1(P-1) through passage-2 (P-2) remained competent to differentiate into mucous and squamous cells when grown in air-liquid interface culture. CONCLUSION: P-2 NHNE cell cultures retained many important features of normal epithelium and were suitable for conducting many studies of upper airway cell biology with an expanded cell pool.