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BACKGROUND: In recent years, identification of differentially expressed genes and sample clustering have become hot topics in bioinformatics. Principal Component Analysis (PCA) is a widely used method in gene expression data. However, it has two limitations: first, the geometric structure hidden in data, e.g., pair-wise distance between data points, have not been explored. This information can facilitate sample clustering; second, the Principal Components (PCs) determined by PCA are dense, leading to hard interpretation. However, only a few of genes are related to the cancer. It is of great significance for the early diagnosis and treatment of cancer to identify a handful of the differentially expressed genes and find new cancer biomarkers. RESULTS: In this study, a new method gLSPCA is proposed to integrate both graph Laplacian and sparse constraint into PCA. gLSPCA on the one hand improves the clustering accuracy by exploring the internal geometric structure of the data, on the other hand identifies differentially expressed genes by imposing a sparsity constraint on the PCs. CONCLUSIONS: Experiments of gLSPCA and its comparison with existing methods, including Z-SPCA, GPower, PathSPCA, SPCArt, gLPCA, are performed on real datasets of both pancreatic cancer (PAAD) and head & neck squamous carcinoma (HNSC). The results demonstrate that gLSPCA is effective in identifying differentially expressed genes and sample clustering. In addition, the applications of gLSPCA on these datasets provide several new clues for the exploration of causative factors of PAAD and HNSC.
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Algoritmos , Bases de Datos Genéticas , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Análisis de Componente Principal , Análisis por Conglomerados , Expresión Génica , Humanos , Neoplasias/genética , Mapas de Interacción de ProteínasRESUMEN
Chronic diseases are known to cause premature aging and frailty. Data about telomere length and telomere length-regulating proteins after pediatric KTx are scarce. Leukocyte telomere length and gene expression level of eight telomere-binding proteins were analyzed in 20 KTx recipients, eight childhood NBL survivors, and nine healthy controls. The influence of key clinical parameters on telomere length and on regulators of telomere length was evaluated. The telomere length in the KTx recipients tended to be shorter (0.53 AU) than in the healthy controls (0.64 AU) but longer than in the NBL survivors (0.38 AU). There was no significant difference in telomere length between the NBL survivors and the KTx recipients (P = .110). The gene expression level of telomere length-preserving protein RPA1 was significantly higher in the KTx recipients than among the NBL survivors or healthy controls, while the expression of TRF2 and the tumor suppressor gene p16 was significantly higher in the KTX recipients when compared to the controls. TRF2 and TIN2 correlated significantly with hsCRP; additionally, TRF2 showed significant correlation with plasma creatinine and eGFR. KTx recipients have near to normal telomere length, but they have significantly higher gene expression levels of telomere regulatory proteins compared with healthy controls, suggesting activation of mechanisms preserving telomere length among KTx recipients. Our results suggest that declined graft function and consequent inflammatory response may have influence on telomerase activity.
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Supervivientes de Cáncer , Trasplante de Riñón , Acortamiento del Telómero , Proteínas de Unión a Telómeros/metabolismo , Adolescente , Adulto , Biomarcadores/metabolismo , Estudios de Casos y Controles , Estudios Transversales , Humanos , Masculino , Adulto JovenRESUMEN
Feature selection and sample clustering play an important role in bioinformatics. Traditional feature selection methods separate sparse regression and embedding learning. Later, to effectively identify the significant features of the genomic data, Joint Embedding Learning and Sparse Regression (JELSR) is proposed. However, since there are many redundancy and noise values in genomic data, the sparseness of this method is far from enough. In this paper, we propose a strengthened version of JELSR by adding the L1-norm constraint on the regularization term based on a previous model, and call it LJELSR, to further improve the sparseness of the method. Then, we provide a new iterative algorithm to obtain the convergence solution. The experimental results show that our method achieves a state-of-the-art level both in identifying differentially expressed genes and sample clustering on different genomic data compared to previous methods. Additionally, the selected differentially expressed genes may be of great value in medical research.
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Algoritmos , Análisis por Conglomerados , Neoplasias del Colon/genética , Bases de Datos como Asunto , Neoplasias Esofágicas/genética , Perfilación de la Expresión Génica , Humanos , Análisis de RegresiónRESUMEN
Point mutations and deletions of mitochondrial DNA (mtDNA) accumulate in tissues during aging in animals and humans and are the basis for mitochondrial diseases. Testosterone synthesis occurs in the mitochondria of Leydig cells. Mitochondrial dysfunction (as induced here experimentally in mtDNA mutator mice that carry a proofreading-deficient form of mtDNA polymerase γ, leading to mitochondrial dysfunction in all cells types so far studied) would therefore be expected to lead to low testosterone levels. Although mtDNA mutator mice showed a dramatic reduction in testicle weight (only 15% remaining) and similar decreases in number of spermatozoa, testosterone levels in mtDNA mutator mice were unexpectedly fully unchanged. Leydig cell did not escape mitochondrial damage (only 20% of complex I and complex IV remaining) and did show high levels of reactive oxygen species (ROS) production (>5-fold increased), and permeabilized cells demonstrated absence of normal mitochondrial function. Nevertheless, within intact cells, mitochondrial membrane potential remained high, and testosterone production was maintained. This implies development of a compensatory mechanism. A rescuing mechanism involving electrons from the pentose phosphate pathway transferred via a 3-fold up-regulated cytochrome b5 to cytochrome c, allowing for mitochondrial energization, is suggested. Thus, the Leydig cells escape mitochondrial dysfunction via a unique rescue pathway. Such a pathway, bypassing respiratory chain dysfunction, may be of relevance with regard to mitochondrial disease therapy and to managing ageing in general.
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Envejecimiento/genética , Células Intersticiales del Testículo/metabolismo , Mitocondrias/genética , Enfermedades Mitocondriales/genética , Envejecimiento/metabolismo , Animales , Citocromos b5/genética , Citocromos b5/metabolismo , Citocromos c/genética , Citocromos c/metabolismo , ADN Mitocondrial/genética , Masculino , Potencial de la Membrana Mitocondrial/genética , Ratones , Ratones Endogámicos C57BL , Mitocondrias/metabolismo , Enfermedades Mitocondriales/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Espermatozoides/metabolismo , Testosterona/genética , Testosterona/metabolismoRESUMEN
BACKGROUND: With increased long-term survivors of childhood cancer patients, therapy-associated infertility has become one of the most common late side-effects and significantly affects their life-quality. Therefore, evaluation of anti-cancer agents on male reproduction and infertility prevention are urgently demanding. The proteasome inhibitor bortezomib has been launched in clinical trials for childhood cancers, however, its potential side effects on reproduction have so far been neither investigated experimentally nor reported in treated children. Thus the present study is designed to explore the impact of bortezomib on male reproductive function and to gain insights into how bortezomib exerts its adverse effects on man gonad, thereby providing pediatric oncologists relevant information. METHODS: 35 day-old male mice were treated with one 11-day cycle of bortezomib and then sacrificed 2 days, 45 days, or 6 months later. A mating study was performed in the group followed for 6 months, and their pups were analyzed on postnatal day 50. Serum follicle-stimulating hormone (FSH) and testicular testosterone levels were measured. Testicular morphology was evaluated by light- and electron microscopy, and the underlying mechanisms and pathways of testis damage were investigated. RESULTS: Testicular damage was visible already 2 days after stopping bortezomib and increased in severity by day 45. Then 80% of seminiferous tubules exhibited hypospermatogenesis with arrest at the levels of spermatogonia, spermatocytes and round spermatids. Germ cells were specifically targeted by bortezomib as evidenced by increased apoptosis mediated through activation of p53 and caspases. Even six months after the bortezomib treatment, testis weight, sperm concentration and seminiferous tubule length remained at a decreased level, indicating that spermatogenesis and tubular outgrowth could not fully recover. Combined with persistently increased serum levels of FSH in these mice, our results demonstrate that bortezomib can have long-term effects on testicular function, although fertility of bortezomib-exposed males remained and their offspring looked healthy. CONCLUSION: Bortezomib treatment causes long-term gonadal dysfunction in male mice. Careful monitoring of gonadal function in male childhood cancer patients treated with bortezomib is thus strongly recommended.
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Ácidos Borónicos/efectos adversos , Pirazinas/efectos adversos , Espermatogénesis/efectos de los fármacos , Enfermedades Testiculares/patología , Testosterona/biosíntesis , Animales , Ácidos Borónicos/administración & dosificación , Bortezomib , Hormona Folículo Estimulante/sangre , Humanos , Masculino , Ratones , Neoplasias/complicaciones , Neoplasias/tratamiento farmacológico , Inhibidores de Proteasoma/administración & dosificación , Pirazinas/administración & dosificación , Enfermedades Testiculares/inducido químicamente , Testículo/efectos de los fármacos , Testículo/patologíaRESUMEN
The coxsackievirus and adenovirus receptor (CXADR (CAR)) is a cell adhesion molecule expressed mainly in epithelial cells. Numerous evidence indicate that CXADR has an important role in testis development and function of the blood-testis barrier (BTB) in vitro. The role of CXADR in testis physiology in vivo has, however, not been addressed. We therefore constructed a conditional CXADR knockout (cKO) mouse model in which CXADR can be depleted at any chosen timepoint by the administration of tamoxifen. We report for the first time that testicular depletion of CXADR in adult and pubertal mice does not alter BTB permeability or germ cell migration across the BTB during spermatogenesis. Adult cKO mice display normal junctional ultra-structure and localization of the junctional proteins claudin-3, occludin, junction-associated molecule-A (JAM-A), and ZO1. The BTB was intact with no leakage of biotin and lanthanum tracers into the tubular lumen. Adult CXADR cKO mice were fertile with normal sperm parameters and litter size. Breeding experiments and genotyping of the pups demonstrated that CXADR-negative sperm could fertilize WT eggs. In addition, knocking down CXADR from postnatal day 9 (P9) does not affect testicular development and BTB formation. These cKO mice were analyzed at P49 and P90 and display an intact barrier and uncompromised fertility. We conclude that CXADR possesses no direct role in testicular physiology in vivo.
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Barrera Hematotesticular/metabolismo , Proteína de la Membrana Similar al Receptor de Coxsackie y Adenovirus/deficiencia , Espermatogénesis , Espermatozoides/metabolismo , Factores de Edad , Animales , Barrera Hematotesticular/ultraestructura , Proteína de la Membrana Similar al Receptor de Coxsackie y Adenovirus/genética , Femenino , Fertilidad , Uniones Intercelulares/metabolismo , Tamaño de la Camada , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Permeabilidad , Embarazo , Maduración Sexual , Proteínas de Uniones Estrechas/metabolismoRESUMEN
Because alternative RNA splicing regulation in the testis is prevalent, we explored testes of Sprague-Dawley rats for existence of alternatively spliced colony-stimulating factor 1 receptor (CSF-1R) mRNA. Using RT-PCR and sequencing, we identified a variant of CSF-1R mRNA that was 284 bp shorter than the full-length CSF-1R transcript. This variant was present in the testis (late fetal stage to adult) and in other organs of rats (7 and 60 days old). The deletion of 284 bp disrupted the open reading frame, resulting in a noncoding mRNA product. When testicular macrophages were stimulated with CSF-1R ligand and lipopolysaccharide, proportionally increased expression of both short isoform and full-length CSF-1R mRNA was observed. Thus, the identified isoform of CSF-1R mRNA may interfere with the expression of full-length CSF-1R mRNA, thereby affecting the biological activity of the ligand/receptor signaling axis in Sprague-Dawley rats.
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ARN Mensajero/metabolismo , Receptor de Factor Estimulante de Colonias de Macrófagos/metabolismo , Testículo/metabolismo , Empalme Alternativo , Animales , Animales Recién Nacidos , Células Cultivadas , Femenino , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Isoformas de Proteínas/genética , Ratas , Ratas Sprague-Dawley , Receptor de Factor Estimulante de Colonias de Macrófagos/genética , Receptor de Factor Estimulante de Colonias de Macrófagos/farmacología , Testículo/embriología , Testículo/crecimiento & desarrolloRESUMEN
Lithium trapping, which is associated with the immobilization of lithium and is one of key factors contributing to structural degradation of lithium-ion batteries during electrochemical cycling, can exacerbate mechanical stress and ultimately cause the capacity loss and battery failure. Currently, there are few studies focusing on how lithium trapping contributes to mechanical stress during electrochemical cycling. This study incorporates the contribution of lithium trapping in the analysis of mechanical stress and mass transport in the framework of finite deformation. Two de-lithiation scenarios are analyzed: one with a constant concentration of trapped lithium and the other with inhomogeneous distribution of trapped lithium. The results show that the constant concentration of trapped lithium increases chemical stress and the inhomogeneous distribution of trapped lithium causes the decrease of chemical stress. The findings can serve as a basis for developing effective strategies to mitigate the lithium trapping and improve the battery performance.
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PURPOSE AND METHODS: To elucidate whether previous cancer treatment affects graft recovery and follicle numbers, morphology, and development in grafts, cryopreserved ovarian biopsies obtained from 18 cancer patients aged 1-24 years with and without exposure to chemotherapy were xenografted as 1 mm3 fragments to immunodeficient mice for 22 weeks with exogenous stimulation. RESULTS: Graft recovery showed no association with chemotherapy exposure, pubertal stage, or leukemia contamination. Total follicle number per recovered graft varied between 0 and 1031 in the chemotherapy-exposed and between 0 and 502 in the non-chemotherapy-exposed group. Atretic follicles formed the largest proportion of the follicle pool in chemotherapy-exposed grafts. Increased atresia correlated with exposure to alkylating agents (mean ± SD 8866.2 ± 9316.3 mg/m2) but not with anthracyclines, pubertal stage, or leukemia contamination. CONCLUSION: The observation confirms the harmful effects of alkylating agents on ovarian tissue. Therapy at the median cumulative dose of 8866 mg/m2 leads to the decreased quality of cryopreserved ovarian follicles in children and young adults.
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Elimination of contaminating malignant cells is crucial to avoiding cancer relapse in association with transplantation of autologous spermatogonial stem cells. In the clinical setting, there is presently no effective procedure for separating testicular cells from cancer cells. Here, CD4, a selective surface marker for Roser's rat leukaemic T-cells, was utilized to eliminate cancer cells from testicular cell samples from leukaemic piebald variegated (PVG) rats by magnetic-activated cell sorting (MACS). All animals receiving MACS-selected testicular cells died within 14-15 days. Only one-third of the contaminating leukaemic cells could be removed from testicular samples. An increase in antibody concentration enhanced the proportion of leukaemic cells removed from 27 to 49%. Variations in the cell size and expression of surface antigens on testicular leukaemic cells were the major obstacles to purification based on this marker. It is concluded that MACS does not prevent transmission of leukaemia to syngenic PVG rats when cells from leukaemic testes are used for testicular cell transplantation.
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Separación Inmunomagnética , Leucemia de Células T/patología , Células Madre Neoplásicas/citología , Testículo/citología , Animales , Citometría de Flujo , Inmunohistoquímica , Masculino , RatasRESUMEN
Childhood cancer treatment can lead to infertility. Organ culture of early postnatal testicular tissue might provide a valuable approach to the study of acute testicular toxicity. The aim of the present study was to develop a functional in vitro organ culture method, in order to identify sensitive target cells to doxorubicin-induced cytotoxicity in immature rat testis during germ cell migration prior initiation of the first wave of spermatogenesis. Testicular tissue fragments from 5-day-old Sprague-Dawley rats were cultured in the absence or presence of doxorubicin (40 and 100ng/ml) and morphology, apoptosis, proliferation and testosterone secretion was analyzed. Postnatal testicular development proceeded normally in control samples for 48h in vitro. In these untreated culture conditions germ and Sertoli cell numbers and germ cell migration were comparable to in vivo. Germ cells were the primary, most sensitive targets for in vitro-induced doxorubicin (100ng/ml) toxicity and their death was not associated with any morphological defects in the Sertoli cells. Organ culture which reduces the need of animal experimentation can be used to study the cytotoxic effects of doxorubicin on the immature testis.
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Alternativas al Uso de Animales , Antibióticos Antineoplásicos/toxicidad , Doxorrubicina/toxicidad , Técnicas de Cultivo de Órganos/métodos , Testículo/efectos de los fármacos , Animales , Animales Recién Nacidos , Apoptosis/efectos de los fármacos , Biomarcadores/metabolismo , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , ADN/biosíntesis , Relación Dosis-Respuesta a Droga , Células Germinativas/efectos de los fármacos , Masculino , Ratas , Ratas Sprague-Dawley , Células de Sertoli/efectos de los fármacos , Testículo/metabolismo , Testículo/patología , Testosterona/metabolismo , Timidina/metabolismoRESUMEN
The noise problem of cancer sequencing data has been a problem that can't be ignored. Utilizing considerable way to reduce noise of these cancer data is an important issue in the analysis of gene co-expression network. In this paper, we apply a sparse and low-rank method which is Robust Principal Component Analysis (RPCA) to solve the noise problem for integrated data of multi-cancers from The Cancer Genome Atlas (TCGA). And then we build the gene co-expression network based on the integrated data after noise reduction. Finally, we perform nodes and pathways mining on the denoising networks. Experiments in this paper show that after denoising by RPCA, the gene expression data tend to be orderly and neat than before, and the constructed networks contain more pathway enrichment information than unprocessed data. Moreover, learning from the betweenness centrality of the nodes in the network, we find some abnormally expressed genes and pathways proven that are associated with many cancers from the denoised network. The experimental results indicate that our method is reasonable and effective, and we also find some candidate suspicious genes that may be linked to multi-cancers.
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Minería de Datos , Redes Reguladoras de Genes/genética , Neoplasias/genética , Bases de Datos Genéticas , Perfilación de la Expresión Génica , Humanos , Análisis de Componente PrincipalRESUMEN
BACKGROUND: Gene co-expression network is a favorable method to reveal the nature of disease. With the development of cancer, the way to build gene co-expression networks based on cancer data has been become a hot spot. However, there are still a limited number of current node measurement methods and node mining strategies for multi-cancers network construction. METHODS: In this paper, we introduce a new method for mining information of co-expression network based on multi-cancers integrated data, named PMN. We construct the network by combining the different types of relevant measures (linear and nonlinear rules) for different nodes based on integrated gene expression data of multi-cancers from The Cancer Genome Atlas (TCGA). For mining genes, we combine different properties (local and global characteristics) of the nodes. RESULTS: We uncover more suspicious abnormally expressed genes and shared pathways of different cancers. And we have also found some proven genes and pathways; of course, there are some suspicious factors and molecules that need clinical validation. CONCLUSIONS: The results demonstrate that our method is very effective in excavating gene co-expression genes of multi-cancers.
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Minería de Datos , Bases de Datos Genéticas , Redes Reguladoras de Genes , Neoplasias/genética , Genes Relacionados con las Neoplasias , HumanosRESUMEN
In a split-litter design experiment, male piglets were exposed orally three times weekly to 300 mg/kg of di(2-ethylhexyl) phthalate (DEHP) or placebo between three and seven weeks of age. The effects on the reproductive organs were examined immediately after the exposure at seven weeks of age in one sub-group, and postpuberally at nine months of age in the other. Morphological features of testes were unaffected at either age group; there were no differences (p>0.05) between the treatments in number of Sertoli cells (as identified by immunostaining with GATA-4 antibodies), percent area of Leydig cells (as detected by 3beta-hydroxysteroid dehydrogenase histochemistry), or incidence of germinal epithelial lesions (histopathology of H&E-stained (hematoxylin and eosin) sections). Three of the seven DEHP-treated animals in seven-week-old group had bulbourethral glands at a stage of maturation far more advanced than that of controls. While there were no obvious differences in the cellular composition between the treatment groups in nine-month-old animals, the bulbourethral glands were heavier (p<0.05) in DEHP-treated boars. Collectively, these features indicate that adolescent exposure to DEHP induces precocious maturity of bulbourethral glands in pigs with persistent effects lasting into adulthood.
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Glándulas Bulbouretrales/efectos de los fármacos , Dietilhexil Ftalato/toxicidad , Enfermedades de los Genitales Masculinos/inducido químicamente , Genitales Masculinos/anatomía & histología , Genitales Masculinos/efectos de los fármacos , Maduración Sexual/efectos de los fármacos , Factores de Edad , Animales , Glándulas Bulbouretrales/patología , Enfermedades de los Genitales Masculinos/patología , Genitales Masculinos/crecimiento & desarrollo , Masculino , Porcinos/anatomía & histología , Porcinos/crecimiento & desarrollo , Factores de TiempoRESUMEN
OBJECTIVE: To observe the effect of acupuncture combined with medication (Rule Granule) on serum prolactin (PRL)ï¼estradiol (E2) and progestone (P) contents and expressions of prolactin receptor (PRLR), estrogen receptor (ER) and progestrone receptor (PR) proteins in mammary gland (MG) tissues of MG hyperplasia (MGH) rats, so as to explore its mechanisms underlying improvement of MGH. METHODS: A total of 55 female SD rats were randomized into blank control group (nï¼10), model group (nï¼10), acupuncture group (nï¼11), medication group (nï¼10), and acupuncture plus medication group (nï¼9). The MGH model was established by muscular injection of E2 benzoate(0.5 mg/kg, once daily for 20 days), followed by injection of P (5 mg/kg) into the lateral muscle of the hind-limb, once daily for 5 days, and the rats of the control group were treated by muscular injection of normal saline at the same site and in the same procedures. Acupoint group A composed of bilateral Tianzhong (SI 11), Ganshu (BL 18) and Zusanli (ST 36), and group B composed of Tanzhong (CV 17), and bilateral Wuyi (ST 15) and Hegu (LI 4) were alternatively punctured with filiform needles and stimulated swiftly by twirling the needle in each acupoint for 20ï¼30 sec, once daily for 30 days. Rats of the medication group and acupuncture ï¼medication group were treated by gavage of Rule Granule fluid containing Baishao (Paeonia Iactiflora Pall), Danggui (Angelica sinensis), etc. (1.5 mL/100 g for each rat) and those of the other groups treated by gavage of distilled water (1.5 mL/100 g). The treatment was given once daily for continuous 30 d. At the end of the experiments, the rats' abdominal aorta blood was collected for assaying the contents of serum PRL, E2 and P with ELISA, and the pathological changes of breast tissue (the left 2nd pairs) were observed under microscope after sectioning and Hï¼E. staining. The expression of PRLR, ER and PR proteins in the breast tissue was detected by immunohistochemistry. RESULTS: After modeling, the hyperplasia of mammary gland in the model group was obvious; the serum PRL and E2 contents and the immunoactivities of PRLR, ER and PR were significantly increased (P<0.01); and the content of serum P was considerably decreased in the model group in comparison with the blank control group (P<0.01). After the treatment, the hyperplasia of mammary gland was improved; the serum PRL and E2 contents and the expression of PRLR, ER and PR were significantly decreased (P<0.05, P<0.01); and the content of P was notably increased in the acupuncture, medication and acupuncture plus medication groups relevant to the model group (P<0.05ï¼P<0.01). The therapeutic effects of acupuncture plus medication were significantly superior to those of both simple medication and simple acupuncture in down-regulating serum PRL and E2 contents and PRLR, ER and PR immunoactivity, as well as in up-regulating serum P content (P<0.05). No significant differences were found between the medication and acupuncture groups in the above mentioned 3 serum and 3 mammary indexes (P>0.05)ï¼. CONCLUSION: Acupuncture, Rule Granule, and acupuncture combined with Rule Granule can improve hyperplasia of mammary gland in rats, which may be related to their effects in reducing serum PRL and E2 and breast ER, PR and PRLR expression levels, and in increasing serum P level. The therapeutic effects of acupuncture plus Rule Granule are better than those of simple acupuncture and simple Rule Granule.
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Terapia por Acupuntura , Animales , Estradiol , Femenino , Hiperplasia , Prolactina , Ratas , Ratas Sprague-DawleyRESUMEN
The underlying primary damage to the seminiferous epithelium caused by chemotherapeutic regimens at childhood is largely unknown. The present investigation was designed to identify acute cytotoxic events in the testis caused by a single dose of doxorubicin. Male rats at 6, 16, and 24 days of age were injected with doxorubicin (3 mg/kg, i.p.) or vehicle (saline) alone and 24 and 48 hours later, the germ cell types and apoptotic cells in the seminiferous epithelium were examined. As indicated by microscopy and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling staining, an 8-fold increase in the number of apoptotic germ cells in the testes of 6-day-old rats was observed 48 hours after doxorubicin treatment. Spermatogonia migrating to the basement membrane were the primary cell type undergoing this induced apoptosis. A single dose of amifostine (200 mg/kg) administered i.p. 15 minutes before injection of doxorubicin provided no protection against this enhanced apoptosis. Under the same conditions, testicular levels of p53 and activated caspase 8 were elevated, whereas the level of murine double minute-2 was lowered. In contrast, doxorubicin treatment did not result in any significant change in the physiologic, stage-specific germ cell apoptosis occurring in the testes of 16- and 24-day-old rats. These observations suggest that the initiation phase of spermatogenesis is highly sensitive to doxorubicin-induced apoptosis. Gonocytes and early spermatogonia are the cell types that are vulnerable to this p53-trigged apoptosis, which results in a decrease in the size of the pool of germ-line stem cells. Amifostine fails to protect the germ cells against this cytotoxic insult.
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Amifostina/farmacología , Antibióticos Antineoplásicos/toxicidad , Apoptosis/efectos de los fármacos , Doxorrubicina/toxicidad , Espermatogonias/efectos de los fármacos , Testículo/efectos de los fármacos , Factores de Edad , Animales , Antibióticos Antineoplásicos/sangre , Doxorrubicina/sangre , Interacciones Farmacológicas , Inyecciones Intraperitoneales , Masculino , Ratas , Ratas Sprague-Dawley , Testículo/citologíaRESUMEN
OBJECTIVE: To observe the effects of acupuncture combined with Rule granule on breast tissue, prolactin(PRL) and prolactin receptor (PRLR) expression in rats with mammary gland hyperplasia (MGH), and to explore its action mechanism to provide reference for clinical treatment of MGH. METHODS: Fifty-five female SD rats were randomly divided into a blank group, a model group, an acupuncture group, a Rule granule group and a combination group. Except the blank group, the rats in the remaining groups were treated with combined stimulation of estrogenic and progestational hormone to establish MGH model. After model establishment, the rats in the acupuncture group were treated with acupuncture at Plan A of "Tianzong" (SI 11), "Ganshu" (BL 18), "Zusanli" (ST 36) and Plan B of "Wuyi" (ST 15), "Hegu" (LI 4), "Danzhong" (CV 17). Each plan was selected for one acupuncture treatment, and two plans were used alternately. The rats in the Rule granule group were treated with oral administration of granule, 3 mL per times. The rats in the combination group were treated with the same Rule granule, followed by acupuncture, once a day. After consecutive 30-day treatment, blood sample was collected from abdominal aorta; ELISA method was applied to measure the contents of PRL; the HE slice of mammary gland was observed under light microscope; the SABC immunohistochemical method was applied to measure the positive expression of PRLR. RESULTS: The morphology of breast tissue in the model group was consistent with MGH. Compared with the blank group, the serum PRL and the expression of PRLR were increased significantly in the model group (both P<0.01). Compared with the model group, the hyperplasia of mammary gland in each treatment group was improved, and serum PRL and expression of PRLR were significantly reduced (P<0.05, P<0.01), which were more significant in the combination group (both P<0.05). CONCLUSION: Acupuncture, Rule granule and its combination could effectively treat MGH, which is likely to reduce the level of serum PRL and inhibit the binding of PRL to PRLR, as a result, the level of E2 is indirectly inhibited, and the hyperplastic mammary gland is recovered. Compared with acupuncture or Rule granule, the combination of both has better overall efficacy.
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Terapia por Acupuntura/métodos , Glándulas Mamarias Animales/metabolismo , Glándulas Mamarias Animales/patología , Prolactina/metabolismo , Receptores de Prolactina/metabolismo , Puntos de Acupuntura , Animales , Femenino , Hiperplasia/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-DawleyRESUMEN
High dimensionality has become a typical feature of biomolecular data. In this paper, a novel dimension reduction method named p-norm singular value decomposition (PSVD) is proposed to seek the low-rank approximation matrix to the biomolecular data. To enhance the robustness to outliers, the Lp-norm is taken as the error function and the Schatten p-norm is used as the regularization function in the optimization model. To evaluate the performance of PSVD, the Kmeans clustering method is then employed for tumor clustering based on the low-rank approximation matrix. Extensive experiments are carried out on five gene expression data sets including two benchmark data sets and three higher dimensional data sets from the cancer genome atlas. The experimental results demonstrate that the PSVD-based method outperforms many existing methods. Especially, it is experimentally proved that the proposed method is more efficient for processing higher dimensional data with good robustness, stability, and superior time performance.
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Algoritmos , Análisis por Conglomerados , Biología Computacional/métodos , Neoplasias , Bases de Datos Genéticas , Perfilación de la Expresión Génica , Humanos , Neoplasias/genética , Neoplasias/metabolismoRESUMEN
Finding robust culture conditions for in vitro maturation (IVM) of male germ cells is still a challenge. Recently, a testis organ culture method, using Knockout Serum Replacement (KSR), was suggested as a promising approach. However, the efficiency of that model is still not optimal. Hence, we have tried to establish the culture conditions in two laboratories, and to improve the reliability of the culture system to generate mature germ cells. Male mice at three days of age were sacrificed. Testes were cut into small pieces which were cultured atop agarose stands, using Minimum Essential Medium alpha supplemented with different supplements; melatonin, Glutamax, and different concentrations of KSR. The results showed that the duration of culture beyond 18 days had an impact on the number of differentiated germ cells. Supplementation with melatonin and Glutamax revealed a positive influence on the efficiency of male germ cell differentiation in vitro. Furthermore, the results confirmed that KSR had a positive effect on germ cell maturation and testosterone production, with a concentration of at least 10%. In conclusion, this study emphasizes the beneficial role of at least 10% KSR in the IVM of germ cells.
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Diferenciación Celular/efectos de los fármacos , Medios de Cultivo/farmacología , Células Germinativas , Melatonina/farmacología , Suero , Testículo , Animales , Células Germinativas/citología , Células Germinativas/metabolismo , Masculino , Ratones , Técnicas de Cultivo de Órganos/métodos , Testículo/citología , Testículo/metabolismoRESUMEN
Reconstitution of telomerase activity by ectopic expression of telomerase reverse transcriptase (hTERT) results in an immortal phenotype in various types of normal human cells, including fibroblasts. Despite lack of transformation characteristics, it is unclear whether hTERT-immortalized cells are physiologically and biochemically the same as their normal counterparts. Here, we compared the gene expression profiles of normal and hTERT-immortalized fibroblasts by using a cDNA microarray containing 20,736 cDNA clones and identified 172 dysregulated genes or expressed sequence tags (ESTs). One of the highly expressed genes in the hTERT-immortalized fibroblasts (hTERT-BJ cells) encodes epiregulin, a potent growth factor. Blockade of epiregulin reduced the growth of hTERT-BJ cells and colony formation of hTERT-transformed fibroblasts. Moreover, inhibition of epiregulin function in immortal hTERT-BJ cells triggered a senescence program. Our results suggest that both activation of telomerase and subsequent induction of epiregulin are required for sustained cell proliferation. Given the significant difference in gene expression profiles between normal and hTERT-immortalized fibroblasts and the close relationship between epiregulin and tumorigenesis, we conclude that hTERT-immortalized cells may not replace their normal counterparts for studies of normal cell biology and that the use of hTERT for expansion of normal human cells for therapeutic purposes must be approached with caution.