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1.
J Med Virol ; 96(7): e29809, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-39016466

RESUMEN

Pancreatic cancer (PC) is a highly aggressive malignancy with a poor prognosis, making early diagnosis crucial for improving patient outcomes. While the gut microbiome, including bacteria and viruses, is believed to be essential in cancer pathogenicity, the potential contribution of the gut virome to PC remains largely unexplored. In this study, we conducted a comparative analysis of the gut viral compositional and functional profiles between PC patients and healthy controls, based on fecal metagenomes from two publicly available data sets comprising a total of 101 patients and 82 healthy controls. Our results revealed a decreasing trend in the gut virome diversity of PC patients with disease severity. We identified significant alterations in the overall viral structure of PC patients, with a meta-analysis revealing 219 viral operational taxonomic units (vOTUs) showing significant differences in relative abundance between patients and healthy controls. Among these, 65 vOTUs were enriched in PC patients, and 154 were reduced. Host prediction revealed that PC-enriched vOTUs preferentially infected bacterial members of Veillonellaceae, Enterobacteriaceae, Fusobacteriaceae, and Streptococcaceae, while PC-reduced vOTUs were more likely to infect Ruminococcaceae, Lachnospiraceae, Clostridiaceae, Oscillospiraceae, and Peptostreptococcaceae. Furthermore, we constructed random forest models based on the PC-associated vOTUs, achieving an optimal average area under the curve (AUC) of up to 0.879 for distinguishing patients from controls. Through additional 10 public cohorts, we demonstrated the reproducibility and high specificity of these viral signatures. Our study suggests that the gut virome may play a role in PC development and could serve as a promising target for PC diagnosis and therapeutic intervention. Future studies should further explore the underlying mechanisms of gut virus-bacteria interactions and validate the diagnostic models in larger and more diverse populations.


Asunto(s)
Heces , Microbioma Gastrointestinal , Metagenómica , Neoplasias Pancreáticas , Viroma , Humanos , Neoplasias Pancreáticas/virología , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/microbiología , Microbioma Gastrointestinal/genética , Metagenómica/métodos , Heces/virología , Heces/microbiología , Virus/aislamiento & purificación , Virus/genética , Virus/clasificación , Metagenoma , Bacterias/aislamiento & purificación , Bacterias/clasificación , Bacterias/genética , Persona de Mediana Edad , Masculino , Femenino , Anciano , Estudios de Casos y Controles
2.
J Pediatr Nurs ; 31(5): e313-22, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27136715

RESUMEN

UNLABELLED: The synthesizing evidence on the effectiveness of using oil massage to promote the growth of infants is still lacking. This paper aims to determine whether oil massage can promote the physical and neurobehavioral growth of infants according to variables and to evaluate whether oil massage is safe for infant skin. ELIGIBILITY CRITERIA: The randomized controlled trials, clinical controlled trials and quasi-experimentally designed trials published prior to or in 2014 were searched according to predetermined inclusion criteria and exclusion criteria in Medline, PubMed, Ovid, the Cochran Library, and Chinese databases, including the China National Knowledge Infrastructure, Wan Fang database and VIP journal integration platform. Besides, the grey lectures were searched as well through Open Grey, GrayLIT Network and Clinical Trials.gov. SAMPLE: Eight studies out of 625 retrieved articles were eligible for inclusion. RESULTS: Oil massage increased the infant weights, lengths and head circumferences. However, it did not promote a significant advantage in neurobehavioral scores or cause a significant risk of adverse skin reactions. IMPLICATIONS: The core mechanisms and standard procedures of oil massage as well as the preferred oil type should be the focus of future nursing practice and research. CONCLUSIONS: Oil massage may effectively improve the physical growth of infants, and it presents a limited risk of adverse skin reactions. However, the relationship between neurodevelopment and oil massage requires further study.


Asunto(s)
Desarrollo Infantil/fisiología , Masaje/métodos , Aceites Volátiles/farmacología , Estatura/fisiología , Peso Corporal/fisiología , China , Ensayos Clínicos Controlados como Asunto , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Estimulación Física/métodos , Ensayos Clínicos Controlados Aleatorios como Asunto
3.
Int J Nurs Pract ; 19(6): 636-43, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24330215

RESUMEN

The aim of this study was to examine the effect of non-pharmacological staged interventions on fatigue and dyspnoea in patients with chronic obstructive pulmonary disease. A randomized controlled trial was conducted with 64 patients in a tertiary hospital in China from 2010 to 2011. Patients were randomly assigned to the control group (n = 32), who received routine care, and the intervention group (n = 32), who received additional non-pharmacological staged interventions. The Multidimensional Fatigue Inventory and the five-grade Medical Research Council dyspnoea scale were used to collect data at baseline and after 6 weeks. Compared with the control group, patients in the intervention group had significantly lower scores on general fatigue (P < 0.001), physical fatigue (P < 0.001), reduced activity (P < 0.001) and reduced motivation (P = 0.03) and had better relief of dyspnoea (P = 0.02). Our study showed that non-pharmacological staged interventions were effective in relieving fatigue and dyspnoea in patients with chronic obstructive pulmonary disease.


Asunto(s)
Disnea , Fatiga , Enfermedad Pulmonar Obstructiva Crónica/terapia , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología
4.
Food Sci Biotechnol ; 32(6): 793-802, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-37041806

RESUMEN

The investigation of the structure and physicochemical properties of starch extracted from a new variety of purple rice was the aim of this study. Starch extracted from a new variety of purple rice named Tianzi No. 1 (PRS) is different in structure and physicochemical properties compared with waxy rice starch (WRS), japonica rice starch (JRS), and indica rice starch (IRS). PRS granules were diversified in shape, and the birefringence of starch particles were clearly observed. Fourier-transform infrared (FT-IR) spectroscopy exhibited the degree of double helix and low short-range order structure of PRS differed from IRS, JRS and WRS. X-ray diffraction analysis shows that PRS presented a typical A-type XRD pattern and possessed lower crystallinity. Based on rheological experiment results, PRS had the highest apparent viscosity, storage modulus (G') and loss modulus (G″). According to textural experiments, PRS gels had higher textural paraments before and after retrogradation. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01205-w.

5.
J Inflamm Res ; 14: 4347-4359, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34539182

RESUMEN

BACKGROUND: Irritable bowel syndrome (IBS) is characterized with abdominal pain, bloating, and changes in bowel habits, and dealing with IBS is still a clinical challenge. The pathogenesis of IBS has been reported to be linked to low-grade mucosal inflammation, and macrophages contribute to the pathological process of this disease. Kurarinone (KAR), a flavanoid derived from Sophora flavescens, has been found medically effective in many inflammatory conditions and cancers. KAR was previously reported to inhibit LPS-induced expression of inflammatory cytokines in macrophages, whether and how KAR regulates the functions of macrophage in IBS remains to be elusive. METHODS: We established a TNBS-induced IBS mouse model, in which KAR was administrated, and mucosal cytokine expression was measured by qRT-PCR. Additionally, mouse macrophages were generated in vitro and their responses to LPS were evaluated by flow cytometry and qRT-PCR. AhR+/+ or AhR-/- macrophages were transferred into DTx-treated CD11b-DTR transgenic mice to investigate the role of AhR in IBS. We collected colonic biopsies and peripheral blood samples from 64 patients with IBS, and analyzed AhR expression by qRT-PCR. RESULTS: We found KAR effectively alleviated visceral hypersensitivity and maintained intestinal barrier functions in mice with IBS. KAR inhibited LPS-induced macrophage activation and expression of pro-inflammatory genes, while increased anti-inflammatory gene expression including IL-10 in an AhR-dependent manner. Using macrophage-depleted mice, we found that chimera mice with AhR-/- macrophages were more susceptible to TNBS-induced IBS and the therapeutic effect of KAR on IBS was significantly impaired in mice with AhR-/- macrophages. Additionally, we found AhR expression in macrophages of IBS patients was associated with the disease severity. CONCLUSION: Our findings provide new evidences that KAR regulates IBS development via macrophage-intrinsic AhR. KAR might show promise as an immunomodulatory therapeutic agent in treating IBS.

6.
Nurse Educ Today ; 59: 16-20, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28917131

RESUMEN

BACKGROUND: Disaster nurse education has received increasing importance in China. Knowing the abilities of disaster response in undergraduate nursing students is beneficial to promote teaching and learning. However, there are few valid and reliable tools that measure the abilities of disaster response in undergraduate nursing students. OBJECTIVES: To develop a self-report scale of self-efficacy in disaster response for Chinese undergraduate nursing students and test its psychometric properties. PARTICIPANTS AND SETTINGS: Nursing students (N=318) from two medical colleges were chosen by purposive sampling. METHODS: The Disaster Response Self-Efficacy Scale (DRSES) was developed and psychometrically tested. Reliability and content validity were studied. Construct validity was tested by exploratory and confirmatory factor analysis. Reliability was tested by internal consistency and test-retest reliability. RESULTS: The DRSES consisted of 3 factors and 19 items with a 5-point rating. The content validity was 0.91, Cronbach's alpha coefficient was 0.912, and the intraclass correlation coefficient for test-retest reliability was 0.953. The construct validity was good (χ2/df=2.440, RMSEA=0.068, NFI=0.907, CFI=0.942, IFI=0.430, p<0.001). CONCLUSIONS: The newly developed DRSES has proven good reliability and validity. It could therefore be used as an assessment tool to evaluate self-efficacy in disaster response for Chinese undergraduate nursing students.


Asunto(s)
Defensa Civil/normas , Psicometría/normas , Autoeficacia , Estudiantes de Enfermería/psicología , Adolescente , China , Defensa Civil/estadística & datos numéricos , Bachillerato en Enfermería/métodos , Femenino , Humanos , Masculino , Psicometría/instrumentación , Psicometría/estadística & datos numéricos , Reproducibilidad de los Resultados , Encuestas y Cuestionarios , Adulto Joven
7.
Biomed Pharmacother ; 95: 1187-1193, 2017 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28931210

RESUMEN

OBJECTIVE: The long non-coding RNA (lncRNA) prostate cancer-associated transcript 1(PCAT-1) has been shown to be dysregulated and exert vital roles in tumorigenesis and progression of various malignancies. However, the precise molecular mechanism in the metastasis and invasion of HCC remain unclear. METHODS: The expression levels of PCAT1 derived from human HCC tissues and cell lines were analyzed through quantitative real-time PCR. QRT-PCR was also applied to detect the expression of HMGB1 and miR-129-5p. Wound healing assay and transwell assays were performed to analyze cell migration and invasion ability. The mRNA levels and protein expression of HMGB1 were detected by western-blotting analysis and immunohistochemistry, respectively. Luciferase assays were used to investigate binding seeds beteen miRNA-129-5p and other transcripts, such as PCAT-1, HMGB1. RESULT: In this study, our founding demonstrated that PCAT-1 was not only aberrantly upregulated in HCC tissues and cell lines, but also associated with TNM stage, metastasis and Histological grade. In vitro, downregulation of PCAT-1 could reduce the invasion and migration of HCC cells. Moreover, our results showed that PCAT-1 could act as an endogenous RNA by directly binding to miR-129-5p. In addition, Luciferase reporter assay and western blotting analyses showed that PCAT-1 repressed inhibitory effect of miR-129-5p and reverse high mobility group box 1 (HMGB1) expression, a target gene of miR-129-5p. CONCLUSION: PCAT-1 functions as competing endogenous RNA (ceRNA) to provide a better understanding for HCC metastasis, and serves as a potential diagnostic and therapeutic target via PCAT-1/miR-129-5p/HMGB1 regulatory crosstalk for the deadly disease.


Asunto(s)
Carcinoma Hepatocelular/genética , Proteína HMGB1/metabolismo , Neoplasias Hepáticas/genética , MicroARNs/metabolismo , ARN Largo no Codificante/metabolismo , Transducción de Señal , Secuencia de Bases , Carcinoma Hepatocelular/patología , Movimiento Celular/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Hepáticas/patología , Masculino , MicroARNs/genética , Persona de Mediana Edad , Invasividad Neoplásica , Metástasis de la Neoplasia , ARN Largo no Codificante/genética , Regulación hacia Arriba/genética
8.
Oncol Lett ; 12(6): 4377-4384, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28101200

RESUMEN

To aim of the present study was to investigate the association between special AT-rich DNA-binding protein-1 (SATB1) expression and liver cancer metastasis. SATB1 mRNA and protein expression in hepatocellular carcinoma tissues was analyzed by immunohistochemistry, and in two hepatocellular cancer cell lines, MHCC-97H (high metastatic potential) and HepG2 (low metastatic potential), by reverse transcription-polymerase chain reaction and western blot analysis. Transwell migration and wound-healing assays were also performed to investigate the metastasis of liver cancer following upregulation or silencing of SATB1 expression. The results revealed that SATB1 expression was significantly higher in hepatocellular carcinoma tissues compared with carcinoma-adjacent tissues. Furthermore, SATB1 expression was correlated with tumor size, differentiation degree, hemorrhage and/or necrosis, invasion and/or metastases and TNM stage. Both the mRNA and protein expression of SATB1 was higher in MHCC-97H cells than HepG2 cells. In addition, the migration capability of MHCC-97H cells was decreased after SATB1 silencing, whereas the migration capability of HepG2 cells was increased after SATB1 upregulation. SATB1 expression was demonstrated to be positively correlated with liver cancer metastasis. These results indicate that liver cancer metastasis is regulated by SATB1 expression. Thus, immunohistochemical SATB1 expression may present an independent risk factor for the metastasis of liver cancer.

9.
Neural Regen Res ; 11(3): 493-501, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-27127492

RESUMEN

Exogenous substance P accelerates wound healing in diabetes, but the mechanism remains poorly understood. Here, we established a rat model by intraperitoneally injecting streptozotocin. Four wounds (1.8 cm diameter) were drilled using a self-made punch onto the back, bilateral to the vertebral column, and then treated using amniotic membrane with epidermal stem cells and/or substance P around and in the middle of the wounds. With the combined treatment the wound-healing rate was 100% at 14 days. With prolonged time, type I collagen content gradually increased, yet type III collagen content gradually diminished. Abundant protein gene product 9.5- and substance P-immunoreactive nerve fibers regenerated. Partial nerve fiber endings extended to the epidermis. The therapeutic effects of combined substance P and epidermal stem cells were better than with amniotic membrane and either factor alone. Our results suggest that the combination of substance P and epidermal stem cells effectively contributes to nerve regeneration and wound healing in diabetic rats.

10.
Mol Med Rep ; 10(3): 1231-6, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24938715

RESUMEN

There is much evidence suggesting that CCL5 is one of the chemoattractant cytokines involved in diabetes mellitus (DM) with diffuse large B­cell lymphoma (DLBCL). However, the pathological impact is unclear. In the current study, in order to improve understanding regarding the role of CCL5 in DM with DLBCL, the expression levels of CCL5 mRNA were examined in normal B cells, human DLBCL cell lines (Ly1, Ly8 and Ly10) and a mouse DLBCL cell line (A20), as well as those in cells cultured with either 5 or 30 mmol/l glucose. A20­CCL5+ (CCL5 overexpression) and A20­CCL5­ (CCL5 knockdown) subclones were obtained through cell transduction with a lentiviral vector, and were subcutaneously injected into BALB/c DM mice and normal mice. Tumor growth was observed by calculating the tumor volume. The results demonstrated that CCL5 mRNA levels in DLBCL cells were significantly higher than those in the normal cells (P<0.05); and levels in DLBCL cells in 30 mmol/l Glu were significantly higher than in those of DLBCL cells in 5 mmol/l Glu (P<0.05). A20­CCL5+ cells led to tumor formation in DM mice compared with A20 and A20­CCL5­ cells. These results indicate that high levels of CCL5 expression may accelerate DLBCL formation in DM.


Asunto(s)
Quimiocina CCL5/genética , Diabetes Mellitus/genética , Regulación Neoplásica de la Expresión Génica , Linfoma de Células B Grandes Difuso/genética , Animales , Línea Celular Tumoral , Quimiocina CCL5/metabolismo , Modelos Animales de Enfermedad , Células HEK293 , Humanos , Linfoma de Células B Grandes Difuso/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , ARN Mensajero/genética , ARN Mensajero/metabolismo
12.
Zhonghua Shao Shang Za Zhi ; 28(1): 25-31, 2012 Feb.
Artículo en Zh | MEDLINE | ID: mdl-22490536

RESUMEN

OBJECTIVE: To observe the effect of sensory neuropeptide substance P combined with epidermal stem cells (ESC) on wound healing and nerve regeneration in diabetic rats. METHODS: ESC that had been isolated from SD rats were identified and cultured in vitro, and they were inoculated onto nourishing layer of amniotic membrane to construct amniotic membrane-ESC. Four full-thickness skin wounds were produced on the back of each of 48 diabetic rats. The resulted 192 wounds were randomly divided into ESC + substance P group, ESC group, substance P group, and control group according to the lottery method, with 48 wounds in each group. Wounds in ESC + substance P group and ESC group were transplanted with amniotic membrane-ESC, and those in substance P group and control group were transplanted with amniotic membrane. After transplantation, 250 µL substance P in the concentration of 1 × 10(-7) mol/L was injected around and into the middle of the wounds in ESC + substance P group and substance P group, 2 times a day, and continued for 4 days, while 250 µL PBS solution was injected in the above-mentioned position in ESC group and control group as control, 2 times a day, and continued for 4 days. On post injury day (PID) 4, 7, 10, 14, 17, and 23, the wound healing rate (with 8 wounds at each time point) was observed and determined, and changes in wound tissue structure were observed with HE staining. On PID 4, 7, and 10, collagen distribution in wound tissue was observed with Masson staining, and type I and type III collagen deposition in wound tissue was respectively observed after immunohistochemical staining. The distribution of protein gene product 9.5 (PGP 9.5) and regeneration of substance P positive nerve fibers in wound tissue were observed with immunohistochemical staining on PID 14 and 23. Data were processed with one-way analysis of variance and t test. RESULTS: (1) The wound healing rate in ESC + substance P group reached 100.0% on PID 14, which was obviously earlier than that in ESC group, substance P group, and control group, healing was respectively observed on PID 17, 17, and 23. The wound healing quality in ESC + substance P group was better than that in the other three groups as shown by HE staining. (2) On PID 10, collagen that was darkly stained and widely distributed was observed in wound tissue of ESC + substance P group and substance P group, while collagen in the other two groups was lightly stained and narrowly distributed. Deposition quantity of type I collagen gradually increased, and that of type III collagen gradually decreased in the wounds of each group over time. On PID 4, 7, and 10, distribution amount of type I collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.72, 118.21, 26.71, P values all below 0.01) and control group (with t value respectively 44.37, 22.76, 30.32, P values all below 0.01), while there was no significance between ESC + substance P group and substance P group. On PID 4, 7, and 10, distribution amount of type III collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.27, 28.68, 14.51, P values all below 0.01) and control group (with t value respectively 35.68, 22.52, 22.24, P values all below 0.01). (3) A large amount of PGP 9.5 and regeneration of substance P positive nerve fibers, and some peripheral nerve fibers in deep wound extending to epidermis were observed in wound tissue of ESC + substance P group and substance P group. A small amount of PGP 9.5 and regeneration of substance P positive nerve fibers without peripheral nerve fibers extending to epidermis were observed in deep wound tissue of ESC group and control group. On PID 14, 23, ratios of area of PGP 9.5 positive nerve fiber in the wounds of ESC + substance P group were (3.86 ± 0.25)% and (7.03 ± 0.28)%, and they were significantly higher than those of ESC group [(1.48 ± 0.30)%, (3.01 ± 0.43)%, with t value respectively 23.95, 30.27, P values all below 0.01] and control group [(1.46 ± 0.23)%, (2.84 ± 0.29)%, with t value respectively 27.35, 40.32, P values all below 0.01]. On PID 14, 23, ratios of substance P positive nerve fiber area in the wounds of ESC + substance P group were (2.01 ± 0.14)% and (1.19 ± 0.11)%, which were obviously higher than those of ESC group [(0.85 ± 0.17)%, (1.34 ± 0.21)%, with t value respectively 20.50, 2.60, P < 0.05 or P < 0.01] and control group [(0.74 ± 0.15)%, (1.30 ± 0.17)%, with t value respectively 23.98, 2.41, P < 0.05 or P < 0.01]. CONCLUSIONS: Joint application of substance P and ESC can effectively promote healing of wound and nerve regeneration in diabetic rats.


Asunto(s)
Diabetes Mellitus Experimental , Regeneración Nerviosa , Células Madre/citología , Sustancia P/farmacología , Cicatrización de Heridas , Animales , Diabetes Mellitus Experimental/patología , Células Epiteliales/citología , Ratas , Ratas Sprague-Dawley , Sustancia P/uso terapéutico
13.
Mol Med Rep ; 4(2): 377-81, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-21468580

RESUMEN

The healing of diabetic wounds represents a formidable clinical challenge, and the molecular mechanisms involved in diabetic wound healing are far from clear. In this study, we investigated the expression of ß-catenin and cyclin D1 in the epidermal stem cells (ESCs) of diabetic rats, and explored whether the reduction of ß-catenin and its downstream target in ESCs, cyclin D1, lead to poor wound healing in diabetes mellitus (DM). We found that, compared to the controls, the ESCs of diabetic rats were markedly reduced, the clone formation efficiency of the ESCs was markedly lower, and the mRNA and protein expression of ß-catenin and cyclin D1 was significantly decreased. These findings suggest that the low expression of ß-catenin and cyclin D1 may reduce the activity of ESCs from diabetic rats, which might be one of the important mechanisms of delayed wound healing in DM.


Asunto(s)
Ciclina D1/metabolismo , Diabetes Mellitus Experimental/metabolismo , Epidermis/patología , Células Madre/metabolismo , beta Catenina/metabolismo , Animales , Western Blotting , Forma de la Célula , Ensayo de Unidades Formadoras de Colonias , Ciclina D1/genética , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patología , Regulación de la Expresión Génica , Inmunohistoquímica , Masculino , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Células Madre/patología , beta Catenina/genética
14.
Zhonghua Shao Shang Za Zhi ; 27(1): 26-31, 2011 Feb.
Artículo en Zh | MEDLINE | ID: mdl-21591337

RESUMEN

OBJECTIVE: To analyze expression characteristics of human skin epidermal stem cell at different developmental stages, and to explore its biological significance. METHODS: Health skin samples from 28-32 w fetuses (F group), 4-12 y children (C group), and 35-55 y adult (A group) were harvested, with 10 cases in each group. Epidermis were separated using trypsin digestion and EDTA, and human epidermal stem cells were isolated and purified with type IV collagen attachment method. The monoclonal antibody of integrin beta1 and keratin 19 were used for detection and identification of epidermal stem cells by immunohistochemical staining. Total RNA was extracted from above cells by Trizol one-step method, and were detected by formaldehyde denaturing agarose gel electrophoresis. Probes were prepared and hybridized into cDNA microarray for scanning fluorescent signals and analysis of images, with two-fold differential expression value for screening. Significantly up/down-regulated genes were selected for verification by real time RT-PCR. RESULTS: By comparing expression profile between A and C groups, a total of 1808 genes with differential expression were detected, including 1089 up-regulated genes and 719 down-regulated genes, and they were classified into 128 categories. Among them, 1462 genes were known (found in GeneBank), 346 genes were unknown. A total of 4534 genes with differential expression were detected between C and F groups, in which 1783 genes were up-regulated and 2751 genes were down-regulated, and they were classified into 216 categories. Among them, 3577 genes were known (found in GeneBank), and 957 genes were unknown. There were 1104 genes with differential expression consistently detected in F, C and A groups, which were classified into 32 categories according to gene function. Among them, 94 genes were consistently up-regulated and 75 genes consistently down-regulated. Test results of real time RT-PCR were in accordance with above-mentioned results. CONCLUSIONS: Gene expression profiles of epidermal stem cells cultured in vitro, harvested from fetuses, children, and adult, exhibit obvious difference. This may be closely related to different stages of proliferation and differentiation of human epidermal stem cell and self-repair ability of wound at different developmental stages.


Asunto(s)
Epidermis/crecimiento & desarrollo , Perfilación de la Expresión Génica , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Células Madre/citología , Adulto , Diferenciación Celular , Niño , Preescolar , Células Epidérmicas , Células Epiteliales/citología , Feto/citología , Regulación del Desarrollo de la Expresión Génica , Humanos , Persona de Mediana Edad , Transcriptoma
15.
Artículo en Zh | MEDLINE | ID: mdl-20187441

RESUMEN

OBJECTIVE: Epidermal stem cells (ESCs) can actively participate in wound healing and enhance reepithelialization. To establish ideal diabetes mellitus (DM) rat models and to investigate the expression of keratin 19 (K19), beta1-integrin, beta-catenin, and proliferating cell nuclear antigen (PCNA) in ESCs of DM rat model, then to study the potential mechanism of difficult recovering wounds of diabetic skin. METHODS: Twenty male SD rats (weighing 250-300 g) were divided into DM group and normal control group randomly (n=10). The DM rat model was made by intraperitoneal injected 65 mg/kg streptozocin (STZ), the normal control group was not treated. At 4 weeks after injection, pancreatic tissue was harvested for HE staining in two groups. The ESCs isolated from full-thickness skins of the back of two group rats were cultured and identified. The 2nd passage of ESCs were obtained for immunocytochemical staining of K 19, beta1-integrin, beta-catenin, and PCNA. Meanwhile, the cell cycle were measured by flow cytometry. The cell colony formation rates were detected after 1 week. RESULTS: The achievement ratio of DM rat model was 90% with good stability. HE staining showed that the number of islet cells significantly decreased with degeneration and necrosis in DM group; the structure of islet cell was clear without degeneration and necrosis in normal control group. The integral absorbance values of positive expression for K19, beta1-integrin, beta-catenin, and PCNA in ESCs of DM group (82.63 +/- 14.77, 21.59 +/- 4.71, 6.49 +/- 6.58, and 90.77 +/- 12.44, respectively) were significantly lower than those in the normal control group (151.24 +/- 42.83, 54.48 +/- 17.43, 116.39 +/- 9.26, and 110.62 +/- 20.67, respectively) (P < 0.01). The clone forming efficiency of ESCs in DM group (6.43% +/- 1.01%) was significantly lower than that in the normal control group (11.37% +/- 1.62%) (P < 0.01). Flow cytometry indicated that 88.89% of cultured ESCs in the DM group were in resting state/pre-DNA-synthetic gap (G0/G1), and the apoptosis rate was 3.98%; 91.50% in the normal control group and the apoptosis rate was 0. CONCLUSION: The DM rat model can be effectively induced by intraperitoneal injected 65 mg/kg STZ. The decreased amount and the low proliferation and differentiation capacity of ESCs may be one of the important mechanisms of difficult recovering wounds of DM rats.


Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Integrina beta1/metabolismo , Queratina-19/metabolismo , beta Catenina/metabolismo , Animales , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Diabetes Mellitus Experimental/patología , Células Epidérmicas , Epidermis/metabolismo , Masculino , Ratas , Ratas Sprague-Dawley , Células Madre/citología , Células Madre/metabolismo , Cicatrización de Heridas
16.
Zhonghua Shao Shang Za Zhi ; 25(4): 261-4, 2009 Aug.
Artículo en Zh | MEDLINE | ID: mdl-19951543

RESUMEN

OBJECTIVE: To establish an effective method of transfecting human marrow mesenchymal stem cells (MSC) with human vascular endothelial growth factor 165 (VEGF 165) gene. METHODS: MSCs isolated and cultured in vitro were divided into transfection group (pShuttle-CMV/VEGF 165 plasmid was transfected into MSCs through liposome-mediating method), empty plasmid group (pShuttle-CMV vehicle was transfected into MSCs as control), liposome group (liposome was transfected into MSCs as control) and control group (normal culture). Expressions of mRNA and protein of MSCs were determined by RT-PCR, enzyme-linked immunosorbent assay and Western Blot. Sensitivity to MSCs on VEGF plasmid transfection was detected by MTT test. RESULTS: Expression level of VEGF 165 gene mRNA in transfection group, empty plasmid group, liposome group, and control group was respectively 0.89 +/- 0.03, 0.34 +/- 0.04, 0.40 +/- 0.03, and 0.30 +/- 0.03, and the difference between transfection group and the other three groups was statistically significant (P < 0.01). Content of VEGF protein in transfection group, empty plasmid group, liposome group, and control group was respectively (778 +/- 35), (543 +/- 24), (561 +/- 28), (571 +/- 23) pg/mL, and the difference between transfection group and the other three groups was statistically significant (P < 0.01). In the transfection group, expression level of VEGF protein peaked on 7(th) day after transfection, which was decreased gradually later. In transfection group, expression level of VEGF 165 protein was obviously higher than that of the other three groups (P < 0.01), and no inhibitory effect of VEGF plasmid transfection on MSCs proliferation was found. CONCLUSIONS: The method for transfecting human VEGF 165 gene into MSCs is established in this research, through which target gene and protein can express effectively.


Asunto(s)
Células de la Médula Ósea/citología , Células Madre Mesenquimatosas/citología , Transfección , Factor A de Crecimiento Endotelial Vascular/genética , Técnicas de Cultivo de Célula , Células Cultivadas , Humanos
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