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1.
Nan Fang Yi Ke Da Xue Xue Bao ; 38(4): 402-408, 2018 Apr 20.
Article in Chinese | MEDLINE | ID: mdl-29735439

ABSTRACT

OBJECTIVE: To investigate the effect of estradiol on the expression of antioxidant enzymes in osteoblasts and its role in postmenopausal osteoporosis. METHODS: Rat models of osteoporosis established by ovariectomy were treated with estradiol for 3 months, and the changes in serum levels of reactive oxygen species (H2O2) and antioxidant enzymes (γ -GCS, GSH-ST and GSH-px) were detected. The effects of estradiol on the expression of γ -GCS mRNA and protein in osteoblast-like cells MC3T3-E1, MG63 and OB were examined with PCR and Western blotting. Using a mRNA microarray, we analyzed the changes in the expressions of 84 antioxidant enzymes in the osteoblast cell line MC3T3-E1 following estradiol treatment, and the enzymes with significant changes were verified by PCR. CCK-8 kit was used to evaluate the effect of estradiol and antioxidant NAC on the proliferation of MC3T3-E1 cells. RESULTS: Rat models of osteoporosis were successfully established with ovariectomy. The osteoporotic rats showed significantly increased serum level of reactive oxygen species (H2O2) and decreased levels of antioxidant enzymes. Estrogen treatment of the osteoporotic rats obviously reversed the phenotype of osteoporosis, lowered serum level of reactive oxygen species, and increased the level of γ -GCS. In MC3T3-E1, MG63 and OB cells, estradiol treatment significantly upregulated the expression levels of γ -GCS mRNA and protein. In MC3T3-E1 cells treated with estrogen, the mRNA chip identified 6 upregulated antioxidant enzymes (Gpx6, Gstk1, Nos2, Prdx2, Ngb and Ccs), and the results of PCR verified that estradiol upregulated Ccs and Ngb mRNAs in MC3T3-E1, MG63 and OB cells. Estradiol and antioxidant NAC obviously promoted the proliferation of MC3T3-E1 cells. CONCLUSION: Estradiol significantly increases the expression of antioxidase γ -Gcs, Ccs and Ngb in osteoblasts in vitro. Postmenopausal osteoporosis is closely related with the increase of reactive oxygen species and the decrease of antioxidant levels. In osteoblasts, estrogen deficiency may increase the level of reactive oxygen species, decrease the level of antioxidant enzymes, activate the oxidative stress cascade, and consequently inhibit the proliferation of osteoblasts to aggravate the condition of osteoporosis.


Subject(s)
Antioxidants/metabolism , Estradiol/pharmacology , Osteoblasts/enzymology , Osteoporosis/enzymology , Oxidative Stress , 3T3 Cells , Animals , Cell Differentiation , Female , Hydrogen Peroxide/metabolism , Mice , Osteoblasts/drug effects , Ovariectomy , Rats , Reactive Oxygen Species/metabolism
2.
Nan Fang Yi Ke Da Xue Xue Bao ; 38(2): 141-147, 2018 Feb 20.
Article in Chinese | MEDLINE | ID: mdl-29502051

ABSTRACT

OBJECTIVE: To establish a method for gene delivery in murine renal tissue using lentivirus vector encoding miR-483-5p. METHODS: Thirty-five C57BL/6J mice were randomly divided into control group, low-dose treatment group (5 µL each kidney) , and high?dose treatment group (20 µL each kidney), and in the latter two groups, the lentivirus vector encoding miR-483-5p were injected in the renal cortex. The tissue samples were collected at 7 and 21 days after the injection. A transgenic mouse model with inducible systemic overexpression of miR-483-5p was established in TG483 mice. The Cre-loxp system was used to create a mouse model with renal tubule-specific expression of miR-483-5p. The levels of BUN in the mice were detected and HE staining and fluorometric TUNEL assay were used to observe the morphological changes of the kidneys; real-time qPCR was used to detect miR-483-5p expression in the renal cortex. RESULTS: The mice with overexpression of miR-483-5p had normal renal function without obvious pathological changes or apoptosis in the renal tissue. Renal cortex injection of 20 µL lentivirus resulted in obviously increased level of miR-483-5p at 21 days (1.2∓0.43 vs 8.6∓1.09, P<0.001). miR-483-5p showed a low expression (0.9∓0.09 vs 1.7∓0.19, P<0.05) in TG483 mice and a high expression in the kidney of the transgenic mice established using the Cre-loxp system (1.6∓1.13 vs 12.36∓3.89, P<0.05). CONCLUSION: The transgenic mice with renal tubule-specific expression of miR-483-5p show normal renal function, and this model facilitates further study of the role of miR-483-5p in the kidney.


Subject(s)
Kidney , MicroRNAs/genetics , Transduction, Genetic , Animals , Apoptosis , Lentivirus , Mice , Mice, Inbred C57BL , Mice, Transgenic
3.
Sci Rep ; 7: 42655, 2017 02 22.
Article in English | MEDLINE | ID: mdl-28225024

ABSTRACT

The small GTPase ras homolog enriched in brain (Rheb) is a downstream target of tuberous sclerosis complex 1/2 (TSC1/2) and an upstream activator of the mechanistic target of rapamycin complex 1 (mTORC1), the emerging essential modulator of M1/M2 balance in macrophages. However, the role and regulatory mechanisms of Rheb in macrophage polarization and allergic asthma are not known. In the present study, we utilized a mouse model with myeloid cell-specific deletion of the Rheb1 gene and an ovalbumin (OVA)-induced allergic asthma model to investigate the role of Rheb1 in allergic asthma and macrophage polarization. Increased activity of Rheb1 and mTORC1 was observed in myeloid cells of C57BL/6 mice with OVA-induced asthma. In an OVA-induced asthma model, Rheb1-KO mice demonstrated a more serious inflammatory response, more mucus production, enhanced airway hyper-responsiveness, and greater eosinophil numbers in bronchoalveolar lavage fluid (BALF). They also showed increased numbers of bone marrow macrophages and BALF myeloid cells, elevated M2 polarization and reduced M1 polarization of macrophages. Thus, we have established that Rheb1 is critical for the polarization of macrophages and inhibition of allergic asthma. Deletion of Rheb1 enhances M2 polarization but decreases M1 polarization in alveolar macrophages, leading to the aggravation of OVA-induced allergic asthma.


Subject(s)
Gene Deletion , Hypersensitivity/genetics , Hypersensitivity/immunology , Myeloid Cells/immunology , Myeloid Cells/metabolism , Ovalbumin/immunology , Ras Homolog Enriched in Brain Protein/genetics , Allergens/immunology , Animals , Asthma/genetics , Asthma/immunology , Asthma/pathology , Bronchoalveolar Lavage Fluid/immunology , Disease Models, Animal , Hypersensitivity/pathology , Macrophage Activation/genetics , Macrophage Activation/immunology , Macrophages/immunology , Macrophages/metabolism , Mechanistic Target of Rapamycin Complex 1/metabolism , Mice , Mice, Inbred C57BL , Protein Binding , Ras Homolog Enriched in Brain Protein/metabolism , Signal Transduction , Th1 Cells/immunology , Th1 Cells/metabolism , Th2 Cells/immunology , Th2 Cells/metabolism
4.
J Zhejiang Univ Sci B ; 6(1): 57-60, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15593394

ABSTRACT

OBJECTIVE: To observe and evaluate a method that is effective and practical for treatment of cerebral palsied (CP) children in China. METHOD: The patient's age and disease type and individual specific conditions were considered in choosing therapy methods accordingly: Chinese herbs, acupuncture, auricular seed pressure, point finger pressing, massage, orthopedic hand manipulation, physiotherapy, occupational therapy, language therapy, etc. Meanwhile we created a new CP treatment model that combines hospitalized treatment with family therapy. RESULTS: The majority of CP patients improved greatly in motor and social adaptation capacities after treatment. Wilcoxon paired rank sum test analysis showed that there were significant differences between the data before and after treatment (P<0.01). CONCLUSION: This combined therapy method, based on traditional Chinese medicine and western medicine plus family supplemental therapy, is an effective and practical treatment strategy for CP children in China.


Subject(s)
Cerebral Palsy/epidemiology , Cerebral Palsy/rehabilitation , Medicine, Chinese Traditional/methods , Adolescent , Child , Child, Preschool , China/epidemiology , Combined Modality Therapy/methods , Combined Modality Therapy/statistics & numerical data , Female , Humans , Infant , Male , Treatment Outcome
5.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 33(5): 449-51, 2004 09.
Article in Chinese | MEDLINE | ID: mdl-15476331

ABSTRACT

OBJECTIVE: To study the mental developments in high risk children and the impact of the high risk factors on neurologic abnormalities, mental defect and long-term outcome. METHODS: The mental development of 122 children who had been exposed to high-risk factors and treated between March 1994 to May 1995 during their newborn periods was evaluated. Gesell development scales were performed when they were at 6 and 12 months old. And Wechsler intelligence scales for children (Chinese version) were performed at 6 approximately 7 years old. RESULTS: The children exposed to hypoglycemia during their newborn period and preterm labor had significantly lower IQ, VIQ and PIQ scores (P <0.05). The other risk factors in order were low birth weight, severe anoxia, asphyxia at birth, erythrocythemia, hyperbilirubinemia. There was significant difference between the children exposed to one risk factor and those exposed to two or more risk factors (P <0.05). And there was significant correlation between developmental assessment at 6 and 12 months and mental development at 6 approximately 7 years old (P<0.01). CONCLUSION: The impact of the high risk factors at birth on children's mental development is not negligible. And the risk of development abnormalities will increase if the children were exposed to multiple risk factors. The evaluation of development at 6 approximately 12 months is of predictive value for long-term outcome.


Subject(s)
Asphyxia Neonatorum/complications , Child Development , Hypoglycemia/complications , Intelligence , Apgar Score , Female , Follow-Up Studies , Humans , Infant , Infant, Newborn , Male , Mental Health , Risk Factors
6.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 31(2): 107-110, 2002 04.
Article in Chinese | MEDLINE | ID: mdl-12539271

ABSTRACT

OBJECTIVE: To identify possible risk factors for cerebral palsy (CP) in children. METHODS: A Population-based survey was conducted (including 92 CP cases) in 66 townships of 15 cities of Zhejiang Province from October to November, 1998. 184 of matched controls were selected for comparison. RESULTS: Factors identified which were statistically significant for risk of subsequent childhood Cerebral Palsy included some neonatal diseases, some maternal diseases, low birth weight (<2500 g), maternal irregular menstruation, toxic, substances during pregnancy, malnutrition during pregnancy,and paternal age. CONCLUSION: Several risk factors for Cerebral Palsy were identified. Their prevention may result in redduction of the incidence of Cerebral Palsy.

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