RESUMO
The wheat aphid Sitobion miscanthi is a dominant and destructive pest in agricultural production. Insecticides are the main substances used for effective control of wheat aphids. However, their extensive application has caused severe resistance of wheat aphids to some insecticides; therefore, exploring resistance mechanisms is essential for wheat aphid management. In the present study, CYP6CY2, a new P450 gene, was isolated and overexpressed in the imidacloprid-resistant strain (SM-R) compared to the imidacloprid-susceptible strain (SM-S). The increased sensitivity of S. miscanthi to imidacloprid after knockdown of CYP6CY2 indicates that it could be associated with imidacloprid resistance. Subsequently, the posttranscriptional regulation of CYP6CY2 in the 3' UTR by miR-3037 was confirmed, and CYP6CY2 participated in imidacloprid resistance. This finding is critical for determining the role of P450 in relation to the resistance of S. miscanthi to imidacloprid. It is of great significance to understand this regulatory mechanism of P450 expression in the resistance of S. miscanthi to neonicotinoids.
Assuntos
Afídeos , Sistema Enzimático do Citocromo P-450 , Resistência a Inseticidas , Inseticidas , MicroRNAs , Neonicotinoides , Nitrocompostos , Neonicotinoides/farmacologia , Nitrocompostos/farmacologia , Animais , Inseticidas/farmacologia , Resistência a Inseticidas/genética , Afídeos/genética , Afídeos/efeitos dos fármacos , MicroRNAs/genética , MicroRNAs/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Imidazóis/farmacologiaRESUMO
Imidacloprid is a neonicotinoid insecticide that efficiently controls piercing-sucking mouthparts pests. However, the impact of low lethal concentration of imidacloprid on key demographic parameters of wheat aphids, Schizaphis graminum (R.) and Rhopalosiphum padi (L.) has been scarcely studied. In this study, we used the age stage, two-sex life table approach to investigate the sublethal effects of imidacloprid on the biological traits of S. graminum and R. padi. Bioassays showed that imidacloprid possesses high toxicity to adult S. graminum and R. padi, with LC50 of 3.59 and 13.78 mg L-1 following 24 h exposure. A low lethal concentration of imidacloprid (LC25) significantly decreased adult longevity and total longevity of progeny generation aphids (F1) of S. graminum. Nevertheless, imidacloprid (LC25) had no significant effects on the fecundity and longevity of directly exposed parental parental S. graminum and R. padi (F0). Our results showed that the low lethal concentration of imidacloprid affected the demographic parameters that ultimately impact on the population of S. graminum. This study provides detailed information about the overall effects of imidacloprid on S. graminum and R. padi that might help to manage these two key pests.
Assuntos
Afídeos , Inseticidas , Animais , Neonicotinoides/toxicidade , Fertilidade , Inseticidas/toxicidadeRESUMO
Castration-resistant prostate cancer (CRPC), especially metastatic castration-resistant prostate cancer (mCRPC) is one of the most prevalent malignancies and main cause of cancer-related death among men in the world. In addition, it is very difficult for clinical treatment because of the natural or acquired drug resistance of CRPC. Mechanisms of drug resistance are extremely complicated and how to overcome it remains an urgent clinical problem to be solved. Thus, a comprehensive and thorough understanding for mechanisms of drug resistance in mCRPC is indispensable to develop novel and better therapeutic strategies. In this review, we aim to review new insight of the treatment of mCRPC and elucidate mechanisms governing resistance to new drugs: taxanes, androgen receptor signaling inhibitors (ARSIs) and poly (ADP-ribose) polymerase (PARP) inhibitors (PARPi). Most importantly, in order to improve efficacy of these drugs, strategies of overcoming drug resistance are also discussed based on their mechanisms respectively.
Assuntos
Neoplasias de Próstata Resistentes à Castração , Masculino , Humanos , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/genética , Neoplasias de Próstata Resistentes à Castração/patologia , Resistencia a Medicamentos Antineoplásicos , Taxoides , Transdução de SinaisRESUMO
Intrauterine growth restriction (IUGR) increases the risk of type 2 diabetes mellitus (T2DM) and metabolic diseases. The pancreas of fetuses with IUGR is usually characterized by pancreatic dysplasia and reduced levels of insulin secretion caused by the diminished replication of ß-cells. Previous studies showed that a low dose of ouabain could reduce the apoptosis of embryonic nephric cells during IUGR and partially restore the number of nephrons at birth. The rescued kidneys functioned well and decreased the prevalence of hypertension. Thus, we hypothesized that ouabain could rescue pancreatic development during IUGR and reduce the morbidity of T2DM and metabolic diseases. Maternal malnutrition was used to induce the IUGR model, and then a low dose of ouabain was administered to rats with IUGR during pregnancy. Throughout the experiment, we monitored the pattern of weight increase and evaluated the metabolic parameters in the offspring in different stages. Male, but not female, offspring in the IUGR group presented catch-up growth. Ouabain could benefit the impaired glucose tolerance of male offspring; however, this desirable effect was eliminated by aging. The insulin sensitivity was significantly impaired in male offspring with IUGR, but it was improved by ouabain, even during old age. However, in the female offspring, low birth weight appeared to be a beneficial factor even in old age; administering ouabain exacerbated these favorable effects. Our data suggested that IUGR influenced glucose metabolism in a sex-specific manner and ouabain treatment during pregnancy exerted strongly contrasting effects in male and female rats.
Assuntos
Diabetes Mellitus Tipo 2 , Retardo do Crescimento Fetal , Gravidez , Feminino , Humanos , Ratos , Animais , Masculino , Retardo do Crescimento Fetal/metabolismo , Ouabaína/farmacologia , Ratos Sprague-Dawley , Diabetes Mellitus Tipo 2/complicações , MetabolomaRESUMO
BACKGROUND: Disruptions of angiogenesis can have a significant effect on the healing of uterine scars. Human endometrial perivascular cells (CD146+PDGFRß+) function as stem cells in the endometrium. Cysteine-rich angiogenic inducer 61 (CYR61) plays an important role in vascular development. The purpose of this study was to observe the effects of the transplantation of human endometrial perivascular cells (En-PSCs) overexpressing CYR61 on structural and functional regeneration in rat models of partial full-thickness uterine excision. METHODS: We first sorted human En-PSCs from endometrial single-cell suspensions by flow cytometry. Human En-PSCs expressing low or high levels of CYR61 were then generated via transfection with a CYR61-specific small interfering ribonucleic acid (si-CYR61) construct or overexpression plasmid. To establish a rat model of uterine injury, a subset of uterine wall was then resected from each uterine horn in experimental animals. Female rats were randomly assigned to five groups, including a sham-operated group and four repair groups that received either PBS loaded on a collagen scaffold (collagen/PBS), En-PSCs loaded on a collagen scaffold (collagen/En-PSCs), En-PSCs with low CYR61 expression loaded on a collagen scaffold (collagen/si-CYR61 En-PSCs), and En-PSCs overexpressing CYR61 loaded on a collagen scaffold (collagen/ov-CYR61 En-PSCs). These indicated constructs were sutured in the injured uterine area to replace the excised segment. On days 30 and 90 after transplantation, a subset of rats in each group was sacrificed, and uterine tissue was recovered and serially sectioned. Hematoxylin and eosin staining and immunohistochemical staining were then performed. Finally, the remaining rats of each group were mated with fertile male rats on day 90 for a 2-week period. RESULTS: Sorted En-PSCs expressed all recognized markers of mesenchymal stem cells (MSCs), including CD10, CD13, CD44, CD73, CD90, and CD105, and exhibited differentiation potential toward adipocytes, osteoblasts, and neuron-like cells. Compared with En-PSCs and En-PSCs with low CYR61 expression, En-PSCs with elevated CYR61 expression enhanced angiogenesis by in vitro co-culture assays. At day 90 after transplantation, blood vessel density in the collagen/ov-CYR61 En-PSCs group (11.667 ± 1.287) was greater than that in the collagen/En-PSCs group (7.167 ± 0.672) (P < 0.05) and the collagen/si-CYR61 En-PSCs group (3.750 ± 0.906) (P < 0.0001). Pregnancy rates differed among groups, from 40% in the collagen/PBS group to 80% in the collagen/En-PSCs group, 12.5% in the collagen/si-CYR61 En-PSCs group, and 80% in the collagen/ov-CYR61 En-PSCs group. In addition, four embryos were evident in the injured uterine horns of the collagen/ov-CYR61 En-PSCs group, while no embryos were identified in the injured uterine horns of the collagen/PBS group. CONCLUSIONS: The results showed that CYR61 plays an important role in angiogenesis. Collagen/ov-CYR61 En-PSCs promoted endometrial and myometrial regeneration and induced neovascular regeneration in injured rat uteri. The pregnancy rate of rats treated with transplantation of collagen/En-PSCs or collagen/ov-CYR61 En-PSCs was improved. Moreover, the number of embryos implantation on the injured area in uterus was increased after transplantation of collagen/ov-CYR61 En-PSCs.
Assuntos
Proteína Rica em Cisteína 61/metabolismo , Células-Tronco Mesenquimais/citologia , Neovascularização Fisiológica/fisiologia , Regeneração/fisiologia , Útero/citologia , Útero/lesões , Útero/metabolismo , Animais , Diferenciação Celular/fisiologia , Células Cultivadas , Cromatografia Líquida , Colágeno/metabolismo , Proteína Rica em Cisteína 61/genética , Endométrio/citologia , Endométrio/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Fertilidade/genética , Fertilidade/fisiologia , Citometria de Fluxo , Imunofluorescência , Humanos , Imuno-Histoquímica , Masculino , Células-Tronco Mesenquimais/metabolismo , Miométrio/citologia , Miométrio/metabolismo , Neovascularização Fisiológica/genética , Gravidez , Ratos , Regeneração/genética , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Severe injuries of the uterus may trigger uterine scar formation, ultimately leading to infertility or obstetrical complications. To date, few methods have adequately solved the problem of collagen deposition in uterine scars. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) have shown great promise in clinical applications. The objective of this study was to investigate the effect of a scaffold/UC-MSCs construct on collagen degradation and functional regeneration in rat uterine scars following full-thickness excision of uterine walls. METHODS: In order to establish a rat model of uterine scars, the uterine wall of approximately 1.0 cm in length and 0.5 cm in width (one-third of the uterine circumference) was excised from each uterine horn. A total of 128 scarred uterine horns from 64 rats were randomly assigned to four groups, including a PBS group (n = 32 uterine horns), scaffold group (n = 32 uterine horns), UC-MSCs group (n = 32 uterine horns) and scaffold/UC-MSCs group (n = 32 uterine horns) to investigate the effect of different treatments on the structure and function of uterine scars. PBS, degradable collagen fibres, UC-MSCs or UC-MSCs mixed with gelatinous degradable collagen fibres were injected into four pre-marked points surrounding each uterine scar, respectively. At days 30 and 60 post-transplantation, a subset of rats (n = 8 uterine horns) from each group was euthanized and serial sections of uterine tissues containing the operative region were prepared. Haematoxylin-eosin staining, Masson's trichrome staining, and immunohistochemical staining for MMP-2, MMP-9, α-SMA and vWF were performed. Finally, another subset of rats (n = 16 uterine horns) from each group was mated with male rats at day 60 post-transplantation and euthanized 18 days after the presence of vaginal plugs to check numbers, sizes and weights of fetuses, as well as sites of implantation. RESULTS: The scaffold/UC-MSCs group exhibited obvious collagen degradation compared with the other three groups. At day 60 post-transplantation, the number of MMP-9-positive cells in the scaffold/UC-MSCs group (25.96 ± 3.63) was significantly higher than that in the PBS group (8.19 ± 1.61, P < 0.01), the scaffold group (7.25 ± 2.17, P < 0.01) and the UC-MSCs group (8.31 ± 2.77, P < 0.01). The pregnancy rate in the scaffold/UC-MSCs group (10/16) was also significantly higher than that in the PBS group (2/16, P < 0.017), the scaffold group (1/16, P < 0.017) and the UC-MSCs group (3/16, P < 0.017). CONCLUSIONS: The scaffold/UC-MSCs system facilitated collagen degradation in uterine scars via upregulation of MMP-9, which was secreted by transplanted UC-MSCs, and promoted regeneration of the endometrium, myometrium and blood vessels in uterine scars. Furthermore, the scaffold/UC-MSCs-treated uterine scars showed nearly complete restoration of receptive fertility.
Assuntos
Cicatriz Hipertrófica/terapia , Colágeno/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Útero/lesões , Cicatrização , Animais , Feminino , Metaloproteinase 9 da Matriz/genética , Células-Tronco Mesenquimais/citologia , Proteólise , Ratos , Ratos Sprague-Dawley , Alicerces Teciduais , Regulação para Cima , Útero/metabolismo , Útero/patologiaRESUMO
STUDY QUESTION: Does the transplantation of adipose-derived stem cells (ADSCs) on soluble collagen scaffolds (collagen/ADSCs) have better therapeutic effect than transplantation of ADSCs alone, to treat premature ovarian insufficiency (POI) in a rat model induced by Tripterygium Glycosides (TG)? SUMMARY ANSWER: The transplantation of collagen/ADSCs increased the short-term retention of ADSCs in ovaries and contributed to long-term restoration of ovarian function, as well as the fertility of rats with TG-induced ovarian damage. WHAT IS KNOWN ALREADY: About 50% of young women in China, who have been treated with TG, have subsequently developed ovarian insufficiency. Rats exhibit similar symptoms to these patients when given an equivalent dose of TG. Transplantation of ADSCs improves ovarian function impaired by chemotherapy in rodent models. STUDY DESIGN, SIZE, DURATION: After the administration of TG, 54 POI model rats were randomly assigned to 4 groups: phosphate buffered saline (PBS) ( ITALIC! n = 14), collagen ( ITALIC! n = 11), ADSCs ( ITALIC! n = 16) and collagen/ADSCs ( ITALIC! n = 13). Seventeen normal rats were assigned as control group. The retention of ADSCs in ovaries was confirmed immediately or at 3, 7, 14 and 28 days after transplantation ( ITALIC! n = 9). Four weeks after transplantation, ovarian function was evaluated from estrous cycle, estradiol level, the follicle number, granulosa cell proliferation and a fertility test. PARTICIPANTS/MATERIALS, SETTING, METHODS: To establish the POI model, rats were administered 60 mg TG/kg/day intragastrically for 50 days. The estrous cycles were assessed by vaginal smear. The concentration of plasma estradiol in diestrus stage was measured using a radioimmunoassay kit. Disordered estrous cycles and low serum estradiol levels indicated the successful establishment of the POI model. Four types of suspensions (PBS, collagen, ADSCs and collagen/ADSCs) were transplanted directly into the core of the ovaries. The short-term retention of ADSCs in ovaries was evaluated by small-animal positron emission tomography images immediately after transplantation of (18)F-Fluorodeoxyglucose ((18)F-FDG) labeled ADSCs. The long-term retention of ADSCs in ovaries was observed by immunohistochemistry after transplantation of green fluorescent protein (GFP)-labeled ADSCs. Serial sections of ovaries were prepared for histological analysis, follicle counting, and immunohistochemistry for Ki67 and Cleaved-Caspase-3. For the assessment of fertility, rats were mated with proven fertile male rats for 10 days. MAIN RESULTS AND THE ROLE OF CHANCE: The (18)F-FDG signal decreased more slowly in ovaries injected with collagen/ADSCs than in ovaries with injected with ADSCs alone. Significantly more GFP-positive cells were observed in ovaries injected with collagen/GFP-ADSCs than in ovaries injected with GFP-ADSCs alone up to 14 days after the injection. However, in both groups very few GFP-positive cells were present at 4 weeks after transplantation. The collagen/ADSCs and ADSCs groups both showed better estrous cycle recovery than the PBS and collagen groups. The estradiol (E2) level in the collagen/ADSCs group was significantly increased compared with that of the PBS group ( ITALIC! P < 0.05). The number of antral follicles in the collagen/ADSCs group and the ADSCs group significantly increased compared with the PBS group ( ITALIC! P < 0.05). The granulosa cell proliferation in the collagen/ADSCs group was better than in the PBS group ( ITALIC! P < 0.01). The mating rates of the collagen/ADSCs group (88.9%) and the ADSCs group (90.9%) were higher than that of PBS group (60%, ITALIC! P < 0.05). The pregnancy rates of the collagen/ADSCs group (77.8%) and the ADSCs group (72.7%) were higher than the PBS group (50%, ITALIC! P < 0.05). LIMITATIONS, REASONS FOR CAUTION: We chose ADSCs for their accessibility, convenience and safety. We did not use other cells or materials for POI treatments to show that the collagen/ADSCs are the most promising materials. WIDER IMPLICATIONS OF THE FINDINGS: Soluble collagen scaffolds may be useful in stem cells transplantation therapy for POI. STUDY FUNDING/COMPETING INTERESTS: This work is supported by grants from the 'Strategic Priority Research Program' of the Chinese Academy of Sciences (XDA01030000); Maternal-Fetal Medicine from Jiangsu Province Health Department of China (XK2011027); Clinical Center of Obstetric, Gynecologic and Genetic Diseases, Nanjing Health Department of Jiangsu Province, China; Fundamental Research Funds for the Central Universities (20620140652). The authors declare no competing financial interests. TRIAL REGISTRATION NUMBER: Not applicable.
Assuntos
Preservação da Fertilidade/métodos , Ovário/fisiologia , Insuficiência Ovariana Primária/terapia , Transplante de Células-Tronco/métodos , Adipócitos , Animais , Técnicas de Cultura de Células , Colágeno , Estradiol/sangue , Ciclo Estral , Feminino , Fertilidade , Citometria de Fluxo , Imuno-Histoquímica , Masculino , Ovário/citologia , Ovário/efeitos dos fármacos , Tomografia por Emissão de Pósitrons , Gravidez , Taxa de Gravidez , Insuficiência Ovariana Primária/induzido quimicamente , Ratos , Ratos Sprague-Dawley , Alicerces TeciduaisRESUMO
Transplacental infection plays a critical role in the reproductive failure induced by porcine reproductive and respiratory syndrome virus (PRRSV), yet exposure of sows and gilts to classical PRRSV generally leads to reproductive failure after 85 days of gestation. We report, for the first time, that the susceptibility of fetuses to highly pathogenic PRRSV (HP-PRRSV) is similar at 60 days and 90 days of gestation. This difference from classical PRRSV may contribute to its high pathogenicity. A field study of the HP-PRRSV vaccine in pregnant sows at mid-gestation should be considered.
Assuntos
Troca Materno-Fetal , Síndrome Respiratória e Reprodutiva Suína/transmissão , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Complicações Infecciosas na Gravidez/veterinária , Animais , Feminino , Feto/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Gravidez , Complicações Infecciosas na Gravidez/virologia , SuínosRESUMO
A variety of diseases may lead to hysterectomies or uterine injuries, which may form a scar and lead to infertility. Due to the limitation of native materials, there are a few effective methods to treat such damages. Tissue engineering combines cell and molecular biology with materials and mechanical engineering to replace or repair damaged organs and tissues. The use of human embryonic stem cells (hESCs) as a donor cell source for the replacement therapy will require the development of simple and reliable cell differentiation protocols. This study aimed at efficiently generating endometrium-like cells from the hESCs and at using these cells with collagen scaffold to repair uterine damage. The hESCs were induced by co-culturing with endometrial stromal cells, and simultaneously added cytokines: epidermal growth factor (EGF), platelet-derived growth factor-b (PDGF-b), and E2. Expression of cell specific markers was analyzed by immunofluorescence and reverse trascription-polymerase chain reaction to monitor the progression toward an endometrium-like cell fate. After differentiation, the majority of cells (>80%) were positive for cytokeratin-7, and the expression of key transcription factors related to endometrial development, such as Wnt4, Wnt7a, Wnt5a, Hoxa11, and factors associated with endometrial epithelial cell function: Hoxa10, Intergrinß3, LIF, ER, and PR were also detected. Then, we established the uterine full-thickness-injury rat models to test cell function in vivo. hESC-derived cells were dropped onto collagen scaffolds and transplanted into the animal model. Twelve weeks after transplantation, we discovered that the hESC-derived cells could survive and recover the structure and function of uterine horns in a rat model of severe uterine damage. The experimental system presented here provides a reliable protocol to produce endometrium-like cells from hESCs. Our results encourage the use of hESCs in cell-replacement therapy for severe uterine damage in future.
Assuntos
Colágeno/farmacologia , Células-Tronco Embrionárias/citologia , Endométrio/citologia , Regeneração/fisiologia , Alicerces Teciduais/química , Animais , Biomarcadores/metabolismo , Bovinos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Forma Celular/efeitos dos fármacos , Células-Tronco Embrionárias/efeitos dos fármacos , Endométrio/transplante , Feminino , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Ratos Sprague-Dawley , Regeneração/efeitos dos fármacosRESUMO
Serious injuries of endometrium in women of reproductive age are often followed by uterine scar formation and a lack of functional endometrium predisposing to infertility or miscarriage. Bone marrow-derived mesenchymal stem cells (BM-MSCs) have shown great promise in clinical applications. In the present study, BM-MSCs loaded onto degradable collagen membranes were constructed. Collagen membranes provided 3-dimmensional architecture for the attachment, growth and migration of rat BM-MSCs and did not impair the expression of the stemness genes. We then investigated the effect of collagen/BM-MSCs constructs in the healing of severe uterine injury in rats (partial full thickness uterine excision). At four weeks after the transplantation of collagen/BM-MSCs constructs, BM-MSCs were mainly located to the basal membrane of regenerative endometrium. The wounded tissue adjacent to collagen/BM-MSCs constructs expressed higher level of bFGF, IGF-1, TGFß1 and VEGF than the corresponding tissue in rats receiving collagen construct alone or in spontaneous regeneration group. Moreover, the collagen/BM-MSCs system increased proliferative abilities of uterine endometrial and muscular cells, facilitated microvasculature regeneration, and restored the ability of endometrium to receive the embryo and support its development to a viable stage. Our findings indicate that BM-MSCs may support uterine tissue regeneration.
Assuntos
Colágeno/química , Transplante de Células-Tronco Mesenquimais/métodos , Engenharia Tecidual , Alicerces Teciduais , Útero/citologia , Útero/lesões , Animais , Células da Medula Óssea/metabolismo , Endométrio/metabolismo , Feminino , Fator de Crescimento Insulin-Like I/metabolismo , Células-Tronco Mesenquimais/metabolismo , Ratos , Ratos Sprague-Dawley , Regeneração , Fator de Crescimento Transformador beta1/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Cicatrização/fisiologiaRESUMO
Serious injuries of uterine which lead to scar formation will finally result in infertility or pregnancy complications. There are few effective methods to treat such damages because of the shortage of native tissues. Vascular endothelial growth factor (VEGF) is important for the formation of new vessels and re-epithelialization of endometrium. Here we produced a collagen-binding VEGF by fusing a collagen-binding domain to the N-terminal of native VEGF. After injection into a rat scarred uterus model (partial of rat uterine horn was excised and left for scar formation) the collagen targeting VEGF promoted remodeling of the scarred uterus including the regeneration of endometrium, muscular cells, and vascularization and improved pregnancy outcomes. Thus, collagen-binding VEGF may be a pragmatic solution for the treatment of severe uterine damages.
Assuntos
Colágeno/química , Engenharia Tecidual/métodos , Útero/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Linhagem Celular Tumoral , Cicatriz/patologia , Endométrio/patologia , Feminino , Humanos , Imuno-Histoquímica/métodos , Neovascularização Patológica , Estrutura Terciária de Proteína , Ratos , Ratos Sprague-Dawley , Regeneração , Útero/citologia , CicatrizaçãoRESUMO
Severe damages of uterine endometrium which prevent embryos from implantation and placentation finally often result in infertility or pregnant complications. There is lack of effective treatments due to the limitation of native materials available and complexity of the function and internal environment of uterus. In the present study, a collagen targeting basic fibroblast growth factor (bFGF) delivery system was constructed by a collagen membrane loaded with bFGF fused a collagen-binding domain (CBD) to the N-terminal which limits the diffusion of bFGF from collagen. We tested the bFGF delivery system in rats under the severe uterine damage model (partial rat uterine horn excision/reconstruction), and found this delivery system improved regeneration abilities of uterine endometrium and muscular cells, improved vascularization, as well as better pregnancy outcomes in rats. Therefore, this targeting delivery system may be an effective strategy for uterine tissue regeneration.
Assuntos
Colágeno/farmacologia , Fator 2 de Crescimento de Fibroblastos/farmacologia , Regeneração/efeitos dos fármacos , Alicerces Teciduais/química , Útero/efeitos dos fármacos , Útero/fisiologia , Animais , Bovinos , Proliferação de Células/efeitos dos fármacos , Colágeno/ultraestrutura , Endométrio/irrigação sanguínea , Endométrio/efeitos dos fármacos , Feminino , Humanos , Imuno-Histoquímica , Antígeno Ki-67/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Gravidez , Resultado da Gravidez , Ligação Proteica/efeitos dos fármacos , Ratos , Útero/patologia , Útero/cirurgia , Fator de von Willebrand/metabolismoRESUMO
OBJECTIVE: To study the mechanisms of the expression of exogenous gene injected via the mammary duct in rat mammary gland tissue. METHODS: Thirty-six SD rats were grouped according to the stages of mammary growth (9 weeks after birth, 7, 12 and 18 days of pregnancy, and 7 and 14 days post partum, 6 rats in each stage). pCE-29 plasmid for highly efficient expression of green fluorescent protein (GFP) and in vivo jetPEITM were mixed and injected into the 5th pair of the mammary glands via the mammary ducts in each rat. The mammary glands were taken to prepare freezing slices 48 h after the injection and observed under fluorescence microscope. HE staining was performed for slices with GFP expression to locate the site of GFP expression by comparison of the images before and after staining. Paraffin sections of the mammary glands with HE staining were also obtained from normal rats in different stages of mammary development. RESULTS: After pCE-29 plasmid injection, GFP expression was observed in the mammary glands of 2 rats in pregnancy day 12 group and in postpartum day 7 group. The green spots shown by fluorescence microscopy were found in the mammary epithelial cells after image comparison. The gland cells undergoing cell division were more numerous in the two stages (pregnancy day 12 and postpartum day 7) than in other stages of mammary development. CONCLUSIONS: In the two stages of pregnancy day 12 and postpartum day 7, exogenous genes injected via the mammary duct can be expressed in the mammary glands. In these stages, the stem cells in the mammary gland epithelia differentiate continuously and the mammary gland epithelial cells flourish under the regulation by hormones to allow better chance for exogenous gene acceptance in the context of active DNA replication in the gland cells. This finding provides important experimental evidence for further studies of mammary gland bioreactor in animals.
Assuntos
Proteínas de Fluorescência Verde/metabolismo , Glândulas Mamárias Animais/metabolismo , Plasmídeos/genética , Animais , Feminino , Técnicas de Transferência de Genes , Proteínas de Fluorescência Verde/genética , Histocitoquímica , Injeções , Microscopia de Fluorescência , Plasmídeos/administração & dosagem , Gravidez , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To establish digitized Virtual Chinese Human Male No.1 (VCH-M1) image dataset with a 0.2-mm equal interval. METHODS: The body of a 24-year-old male was used for this study. Perfusion with phenol and vermilion of the arteries was performed, followed by body shape adjustment by cold saline and pre-embedding with broken ices in an upside-down position, which was completed in a stepwise procedure to minimize body shape deformation. Section milling was conducted subsequently with the section thickness of 2 mm and the section images were captured by digital camera, which were immediately transferred to a computer for storage and processing. RESULTS: A total of 9 232 sections were obtained for the whole body, and the resolution of each of the image in TIF format was 3 024x2 016 pixels, resulting in the size of approximately 18 M for each image and about 161 G for the whole dataset. CONCLUSIONS: Compared with VCH-F1, the image quality in VCH-M1 dataset is significantly improved, demonstrated by much clearer tissue boundary in the images and minimized body shape deformation during the embedding process.
Assuntos
Anatomia Transversal , Interface Usuário-Computador , Adulto , China , Coleta de Dados , Humanos , MasculinoRESUMO
Manipulations are essential to the acupuncture curative effect. With the technique limited, the researches on acupuncture manipulations can only be carried out with respect to the senses and experience of doctors and patients, thus seriously impeding the modernization and internationalization of acupuncture. Both the modern integration technology of transducer and the biomechanical principles are applied to develop a detection system that can measure the interaction force between the manipulator and the manipulated patient on the needle at each kind of manipulation. Through clinical practice, the force waveforms acting on needle with the manipulations of symmetrical twirling-rotating and symmetrical lifting-inserting were recorded so as to realize the quantitative, objective and real-time detection of the force during the acupuncture process, which provided a new experimentation means and analysis method for the improvement of clinical curative effect and quantitative research of acupuncture and meridians.