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1.
Indian J Pathol Microbiol ; 2023 Jun; 66(2): 372-374
Article | IMSEAR | ID: sea-223454

ABSTRACT

Herein we present a case of a 62-year-old male patient who was admitted with the chief complaints of nasal obstruction. The histopathological and immunohistochemical evaluation led to a diagnosis of olfactory neuroblastoma with rhabdomyoblasts. A review of the literature revealed that this is only the fourth case of olfactory neuroblastoma with rhabdomyoblasts. Thus, investigation of more cases and longer follow-up is necessary to understand the disease and identify the best treatment to improve prognosis.

2.
Tropical Biomedicine ; : 455-461, 2022.
Article in English | WPRIM | ID: wpr-961206

ABSTRACT

@#Chigger mites are arthropods and are the sole vectors of scrub typhus, and rodents as well as other small mammals are the most common hosts of chigger mite larvae. Therefore, it is of great medical significance to study the ecology of chigger mites. In this study, a detailed analysis of chigger mites was conducted based on field survey data. A total of 4,941 chigger mites were collected from 86 hosts at 34 survey sites in Ruili, Yunnan Province, China. Among the 4,941 chiggers, five genera in one subfamily were identified; Schoengastiella ligula was the dominant chigger species with the highest infestation index, prevalence (Pm, 42.86%) and mean intensity (MI, 59.09%) (P<0.001). The association coefficient (V) between S. ligula and Gahrliepia radiopunctata was positively correlated (P<0.05), indicating the tendency of chiggers to select and coexist on the same host at the same time. The dominant species Leptotrombidium kunmingense, Ascoschoengastia indica, S. ligula and G. radiopunctata showed aggregation distribution patterns, indicating that the distribution of chiggers among different hosts was not uniform. Low altitudes and low latitudes appeared to be more favorable for the growth and reproduction of chigger mites (P<0.05). It is suggested to collect as many host samples as possible in future field investigations to better understand the dynamics of chigger mite populations and their primary hosts.

3.
Tropical Biomedicine ; : 489-498, 2020.
Article in English | WPRIM | ID: wpr-823257

ABSTRACT

@#Toxoplasma gondii, a ubiquitous pathogen that infects nearly all warm-blooded animals and humans, can cause severe complications to the infected people and animals as well as serious economic losses and social problems. Here, one local strain (TgPIG-WH1) was isolated from an aborted pig fetus, and the genotype of this strain was identified as ToxoDB #3 by the PCR RFLP typing method using 10 molecular markers (SAG1, SAG2, alternative SAG2, SAG3, BTUB, GRA6, L358, PK1, C22-8, C29-2 and Apico). A comparison of the virulence of this isolate with other strains in both mice and piglets showed that TgPIG-WH1 was less virulent than type 1 strain RH and type 2 strain ME49 in mice, and caused similar symptoms to those of ME49 such as fever in piglets. Additionally, in piglet infection with both strains, the TgPIG-WH1 caused a higher IgG response and more severe pathological damages than ME49. Furthermore, TgPIG-WH1 caused one death in the 5 infected piglets, whereas ME49 did not, suggesting the higher virulence of TgPIG-WH1 than ME49 during piglet infection. Experimental infections indicate that the virulence of TgPIG-WH1 relative to ME49 is weaker in mice, but higher in pigs. This is probably the first report regarding a ToxoDB #3 strain from pigs in Hubei, China. These data will facilitate the understanding of genetic diversity of Toxoplasma strains in China as well as the prevention and control of porcine toxoplasmosis in the local region.

4.
Braz. j. med. biol. res ; 51(2): e6793, 2018. graf
Article in English | LILACS | ID: biblio-889023

ABSTRACT

Colon cancer is one of the most common digestive tumors. The present study aimed to explore the functional role, as well as the underlying mechanism of long non-coding RNA LINC00261 in colon cancer. Expression of LINC00261 was analyzed in colon cancer cell lines and human normal cell lines. Acquired resistance cell lines were then built and the acquired resistance efficiency was detected by evaluating cell viability. Thereafter, the effects of LINC00261 overexpression on cisplatin-resistant colon cancer cells were measured, as well as cell apoptosis, viability, migration, and invasion. Subsequently, we investigated the interaction of LINC00261 and β-catenin. The results showed that the LINC00261 gene was down-regulated in colon cancer cell lines and tissues, and in cisplatin-resistant cells. LINC00261 overexpression might relieve cisplatin resistance of colon cancer cells via promoting cell apoptosis, and inhibiting cell viability, migration, and invasion. Moreover, LINC00261 might down-regulate nuclear β-catenin through restraining β-catenin from cytoplasm into nuclei or it could also promote β-catenin degradation and inhibit activation of Wnt pathway. Finally, LINC00261 reduced cisplatin resistance of colon cancer in vivo and enhanced the anti-colon cancer effect of cisplatin through reducing tumor volume and weight.


Subject(s)
Humans , RNA, Long Noncoding/physiology , Antineoplastic Agents/pharmacology , Tetrazolium Salts , Thiazoles , Down-Regulation , Blotting, Western , Reproducibility of Results , Analysis of Variance , Apoptosis/drug effects , Apoptosis/physiology , Apoptosis Regulatory Proteins/drug effects , beta Catenin/physiology , Cell Migration Assays
5.
Braz. j. med. biol. res ; 50(2): e5637, 2017. tab, graf
Article in English | LILACS | ID: biblio-839249

ABSTRACT

Immune thrombocytopenia (ITP) is a disease characterized by isolated thrombocytopenia. Abnormal effector T cell activation is an important mechanism in the pathogenesis of ITP. Regulatory T cells (Treg) have a strong immunosuppressive function for T cell activation and their importance in the pathophysiology and clinical treatment of ITP has been confirmed. Myeloid-derived suppressor cells (MDSCs) are other immunosuppressive cells, which can also suppress T cell activation by secreting arginase, iNOS and ROS, and are essential for Treg cells’ differentiation and maturation. Therefore, we speculate that MDSCs might also be involved in the immune-dysregulation mechanism of ITP. In this study, we tested MDSCs and Treg cells in peripheral blood samples of twenty-five ITP patients and ten healthy donors. We found that MDSCs and Treg cells decreased simultaneously in active ITP patients. Relapsed ITP patients showed lower MDSCs levels compared with new patients. All patients received immunosuppressive treatment including dexamethasone alone or in combination with intravenous immune globulin. We found that MDSCs’ level after treatment correlated with platelet recovery. Our study is the first that focused on MDSCs’ role in ITP. Based on our results, we concluded that circulating MDSCs could predict disease activity and treatment response in ITP patients. This preliminary conclusion indicates a substantial significance of MDSCs in the pathophysiology and clinical treatment of ITP, which deserves further investigation.


Subject(s)
Humans , Male , Female , Adult , Myeloid-Derived Suppressor Cells/immunology , Purpura, Thrombocytopenic, Idiopathic/immunology , T-Lymphocytes, Regulatory/immunology , Case-Control Studies , Dexamethasone/administration & dosage , Immunoglobulins, Intravenous/administration & dosage , Lymphocyte Activation , Purpura, Thrombocytopenic, Idiopathic/drug therapy , Purpura, Thrombocytopenic, Idiopathic/physiopathology , T-Lymphocytes, Regulatory/physiology
6.
Indian J Cancer ; 2015 Dec; 52(6)Suppl_2: s119-s124
Article in English | IMSEAR | ID: sea-169287

ABSTRACT

BACKGROUND: Protein kinase CK2 is widely expressed in eukaryotic cells, and plays an important role in cell proliferation, migration, apoptosis, etc. The aim of the current study is to explore how Quinalizarin, a specific CK2 inhibitor, affects the cell proliferation, migration, and apoptosis of different pathological and genetic types of human lung cancer cell lines. MATERIALS AND METHODS: MTT assays were performed to evaluate the cell viability after being treated by Quinalizarin. Transwell migration assays were used to assess whether Quinalizarin could suppress cell migration. Flow cytometry was employed to test the apoptosis rate of different cells. RESULTS: After being treated by Quinalizarin, the viability of different pathological types of lung cancer cells (H446, H460, A549) were significantly suppressed in a time and dose‑dependent manner. More interestingly, in a serial of human lung adenocarcinoma cell lines with different epidermal growth factor receptor (EGFR) mutation status, Quinalizarin was shown to have a much better ability to reduce the viability of cells with EGFR sensitive mutation than those with resistance mutations. Meanwhile, we also found that the cell migration of different pathological types of lung cancer cells (H446, H460, A549) was significantly decreased by Quinalizarin dose‑dependently. In addition, the apoptosis rates in those cells were proved to be increased after exposed to Quinalizarin. CONCLUSIONS: Quinalizarin, the specific CK2 inhibitor, could reduce cell viability with emphasis on adenocarcinoma cells harboring EGFR sensitive mutation, suppresses migration, and accelerates apoptosis in different human lung cancer cell lines.

7.
Braz. j. med. biol. res ; 48(7): 654-664, 07/2015. graf
Article in English | LILACS | ID: lil-751344

ABSTRACT

Recent evidence indicates that a deficiency of 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) may influence asthma pathogenesis; however, its roles in regulating specific molecular transcription mechanisms remain unclear. We aimed to investigate the effect of 1,25(OH)2D3 on the expression and enzyme activity of histone deacetylase 2 (HDAC2) and its synergistic effects with dexamethasone (Dx) in the inhibition of inflammatory cytokine secretion in a rat asthma model. Healthy Wistar rats were randomly divided into 6 groups: control, asthma, 1,25(OH)2D3 pretreatment, 1,25(OH)2D3 treatment, Dx treatment, and Dx and 1,25(OH)2D3 treatment. Pulmonary inflammation was induced by ovalbumin (OVA) sensitization and challenge (OVA/OVA). Inflammatory cells and cytokines in the bronchoalveolar lavage (BAL) fluid and histological changes in lung tissue were examined. Nuclear factor kappa B (NF-κB) p65 and HDAC2 expression levels were assessed with Western blot analyses and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Enzyme activity measurements and immunohistochemical detection of HDAC2 were also performed. Our data demonstrated that 1,25(OH)2D3 reduced the airway inflammatory response and the level of inflammatory cytokines in BAL. Although NF-κB p65 expression was attenuated in the pretreatment and treatment groups, the expression and enzyme activity of HDAC2 were increased. In addition, 1,25(OH)2D3 and Dx had synergistic effects on the suppression of total cell infusion, cytokine release, and NF-κB p65 expression, and they also increased HDAC2 expression and activity in OVA/OVA rats. Collectively, our results indicated that 1,25(OH)2D3 might be useful as a novel HDAC2 activator in the treatment of asthma.


Subject(s)
Animals , Male , Asthma/drug therapy , Calcitriol/pharmacology , /drug effects , NF-kappa B/drug effects , Vitamins/pharmacology , Asthma/chemically induced , Blotting, Western , Bronchoalveolar Lavage Fluid/chemistry , Cell Count , Calcitriol/therapeutic use , Cytokines/analysis , Cytokines/drug effects , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation, Enzymologic/drug effects , /metabolism , Immunohistochemistry , Lung/chemistry , Lung/drug effects , NF-kappa B/analysis , Ovalbumin , Rats, Wistar , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Treatment Outcome , Vitamins/therapeutic use
8.
Braz. j. med. biol. res ; 47(9): 738-745, 09/2014. graf
Article in English | LILACS | ID: lil-719313

ABSTRACT

Hypoxia-inducible factor-1α (HIF-1α) is one of the most potent angiogenic growth factors. It improves angiogenesis and tissue perfusion in ischemic skeletal muscle. In the present study, we tested the hypothesis that ischemic postconditioning is effective for salvaging ischemic skeletal muscle resulting from limb ischemia-reperfusion injury, and that the mechanism involves expression of HIF-1α. Wistar rats were randomly divided into three groups (n=36 each): sham-operated (group S), hindlimb ischemia-reperfusion (group IR), and ischemic postconditioning (group IPO). Each group was divided into subgroups (n=6) according to reperfusion time: immediate (0 h, T0), 1 h (T1), 3 h (T3), 6 h (T6), 12 h (T12), and 24 h (T24). In the IPO group, three cycles of 30-s reperfusion and 30-s femoral aortic reocclusion were carried out before reperfusion. At all reperfusion times (T0-T24), serum creatine kinase (CK) and lactate dehydrogenase (LDH) activities, as well as interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α) concentrations, were measured in rats after they were killed. Histological and immunohistochemical methods were used to assess the skeletal muscle damage and HIF-1α expression in skeletal muscle ischemia. In groups IR and IPO, serum LDH and CK activities and TNF-α, IL-6, and IL-10 concentrations were all significantly increased compared to group S, and HIF-1α expression was up-regulated (P<0.05 or P<0.01). In group IPO, serum LDH and CK activities and TNF-α and IL-6 concentrations were significantly decreased, IL-10 concentration was increased, HlF-1α expression was down-regulated (P<0.05 or P<0.01), and the pathological changes were reduced compared to group IR. The present study suggests that ischemic postconditioning can reduce skeletal muscle damage caused by limb ischemia-reperfusion and that its mechanisms may be related to the involvement of HlF-1α in the limb ischemia-reperfusion injury-triggered inflammatory response.


Subject(s)
Animals , Male , Extremities/blood supply , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Ischemic Postconditioning , Muscle, Skeletal/blood supply , Reperfusion Injury/prevention & control , Blotting, Western , Creatine Kinase/metabolism , Disease Models, Animal , Endothelial Cells/pathology , Immunohistochemistry , /blood , /blood , L-Lactate Dehydrogenase/metabolism , Muscle, Skeletal/injuries , Random Allocation , Rats, Wistar , Time Factors , Tumor Necrosis Factor-alpha/blood , Up-Regulation
9.
Braz. j. med. biol. res ; 47(4): 299-306, 8/4/2014. graf
Article in English | LILACS | ID: lil-705762

ABSTRACT

Notch signaling plays a vital role in tumorigenicity and tumor progression by regulating proliferation, invasion, and the tumor microenvironment. Previous research by our group indicated that Notch ligand Delta-like 1 (Dll1) is involved in angiogenesis in melanoma, and we noticed that it took a longer time to trypsinize Dll1-expressing B16 melanoma cells than the control cells. In this article, we extended our study to investigate the effects of Dll1 on tumor cell adhesion and metastasis. Dll1 overexpression activated Notch signaling in B16 tumor cells and significantly enhanced the adhering capacity of B16 tumor cells both in vitro and in vivo. B16-Dll1 cells also had a higher metastatic potential than their counterpart in the mouse model of lung metastasis. Along with increased Dll1 expression, N-cadherin, but not E-cadherin, was upregulated in B16-Dll1 cells. These data suggested that Notch ligand Dll1 may enhance the adhesion and metastasis of melanoma cells by upregulation of N-cadherin.


Subject(s)
Animals , Cadherins/biosynthesis , Intercellular Signaling Peptides and Proteins/metabolism , Lung Neoplasms/secondary , Melanoma, Experimental/secondary , Membrane Proteins/metabolism , Signal Transduction/genetics , Blotting, Western , Cell Adhesion , Cell Culture Techniques , Cell Line, Tumor , Gene Expression , Green Fluorescent Proteins , Human Umbilical Vein Endothelial Cells/physiology , Melanoma, Experimental/pathology , Real-Time Polymerase Chain Reaction , Up-Regulation
10.
Indian J Cancer ; 2014 Mar; 51(7_Suppl): s103-s105
Article in English | IMSEAR | ID: sea-158233

ABSTRACT

BACKGROUND: Bevacizumab, a recombinant humanized monoclonal antibody that blocks angiogenesis by inhibiting vascular endothelial growth factor A, was described to be effective in the treatment of recurrent or platinum‑resistance ovarian cancer. The present retrospective study was performed to further evaluate the clinical efficacy and toxicity of bevacizumab in the treatment of Chinese recurrent ovarian cancer patients who had been previously treated by platinum‑based chemotherapy. MATERIALS AND METHODS: We reviewed the hospital database and finally included 26 recurrent ovarian cancer patients who were treated with bevacizumab combined with gemcibabine or paclitaxel or single agent. All included patients received >3 cycle of bevacizumab treatment. The tumor response, overall survival, and toxicities were documented. RESULTS: Under the treatment of bevacizumab combined with gemcibabine or paclitaxel, 2 complete response (7.7%), 8 partial response (30.8%), 7 stable disease (26.9%) and 9 progression disease (34.6%) was documented with the objective response rate of 38.5% and disease control rate of 65.4%. The median overall survival from the first application of bevacizumab was 15.3 months [Figure 1] for all of the 26 patients. The median overall survival time was 16.2 and 14.0 months for bevacizumab + gemcitabine and bevacizumab + paclitaxel treatment schedule respectively. The overall survival was not different between bevacizumab + gemcitabine and bevacizumab + paclitaxel treatment regimen hazard ratio = 0.80 (95% confidence interval: 0.32–2, P = 0.64). The hypertension and proteinuria were the major bevacizumab related toxicities. CONCLUSIONS: Bevacizumab combined with gemcibabine or paclitaxel was a promising treatment schedule for platinum‑resistance recurrent ovarian cancer.

11.
Braz. j. med. biol. res ; 46(9): 739-745, 19/set. 2013. tab, graf
Article in English | LILACS | ID: lil-686570

ABSTRACT

Liver cirrhosis is one of the most common diseases of Chinese patients. Herein, we report the high expression of a newly identified histone 3 lysine 4 demethylase, retinoblastoma binding protein 2 (RBP2), and its role in liver cirrhosis in humans. The siRNA knockdown of RBP2 expression in hepatic stellate cells (HSCs) reduced levels of α-smooth muscle actin (α-SMA) and vimentin and decreased the proliferation of HSCs; and overexpression of RBP2 increased α-SMA and vimentin levels. Treatment with transforming growth factor β (TGF-β) upregulated the expression of RBP2, α-SMA, and vimentin, and the siRNA knockdown of RBP2 expression attenuated TGF-β-mediated upregulation of α-SMA and vimentin expression and HSC proliferation. Furthermore, RBP2 was highly expressed in cirrhotic rat livers. Therefore, RBP2 may participate in the pathogenesis of liver cirrhosis by regulating the expression of α-SMA and vimentin. RBP2 may be a useful marker for the diagnosis and treatment of liver cirrhosis.


Subject(s)
Animals , Humans , Male , Actins/metabolism , Hepatic Stellate Cells/metabolism , Histone Demethylases/metabolism , Liver Cirrhosis/metabolism , /metabolism , Vimentin/metabolism , Blotting, Western , Cell Proliferation , Disease Models, Animal , Gene Expression , Gene Knockdown Techniques , Rats, Wistar , RNA, Small Interfering/metabolism , Transforming Growth Factor beta/metabolism
12.
Braz. j. med. biol. res ; 43(5): 451-459, May 2010. tab, ilus
Article in English | LILACS | ID: lil-546337

ABSTRACT

Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures) was enhanced to 60 percent of a total of 10 cultures (initiated from 8 distinct fetal small intestines), allowing the generation of viable epithelial cell cultures. Western blot, real-time PCR and immunofluorescent staining showed that cytokeratins 8, 18 and mouse intestinal mucosa-1/39 had high expression levels in human intestinal epithelial cells. Differentiated markers such as sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV also showed high expression levels in human intestinal epithelial cells. Differentiated human intestinal epithelial cells, with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa-1/39) and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV), may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. This is relatively simple, requiring no sophisticated techniques or instruments, and may have a number of varied applications.


Subject(s)
Humans , Cell Culture Techniques/methods , /pharmacology , Epithelial Cells/cytology , Intestinal Mucosa/cytology , Intestine, Small/cytology , Thermolysin/pharmacology , Cell Differentiation , Cell Line , Cell Movement , Cell Proliferation , Epithelial Cells/drug effects , Fetus , Intestinal Mucosa/embryology , Intestine, Small/embryology
13.
Braz. j. med. biol. res ; 43(4): 330-337, Apr. 2010. ilus, graf
Article in English | LILACS | ID: lil-543582

ABSTRACT

The objectives of this study were to determine the effect of tumor necrosis factor alpha (TNF-á) on intestinal epithelial cell permeability and the expression of tight junction proteins. Caco-2 cells were plated onto Transwell® microporous filters and treated with TNF-á (10 or 100 ng/mL) for 0, 4, 8, 16, or 24 h. The transepithelial electrical resistance and the mucosal-to-serosal flux rates of the established paracellular marker Lucifer yellow were measured in filter-grown monolayers of Caco-2 intestinal cells. The localization and expression of the tight junction protein occludin were detected by immunofluorescence and Western blot analysis, respectively. SYBR-Green-based real-time PCR was used to measure the expression of occludin mRNA. TNF-á treatment produced concentration- and time-dependent decreases in Caco-2 transepithelial resistance and increases in transepithelial permeability to the paracellular marker Lucifer yellow. Western blot results indicated that TNF-á decreased the expression of phosphorylated occludin in detergent-insoluble fractions but did not affect the expression of non-phosphorylated occludin protein. Real-time RT-PCR data showed that TNF-á did not affect the expression of occludin mRNA. Taken together, our data demonstrate that TNF-á increases Caco-2 monolayer permeability, decreases occludin protein expression and disturbs intercellular junctions.


Subject(s)
Humans , Cell Membrane Permeability/drug effects , Epithelial Cells/drug effects , Intestinal Mucosa/cytology , Membrane Proteins/drug effects , Tight Junctions/drug effects , Tumor Necrosis Factor-alpha/pharmacology , Blotting, Western , Epithelial Cells/metabolism , Membrane Proteins/metabolism , Polymerase Chain Reaction/methods , RNA, Messenger/genetics , RNA, Messenger/metabolism , Tight Junctions/metabolism
14.
Braz. j. med. biol. res ; 42(6): 506-514, June 2009. ilus, tab, graf
Article in English | LILACS | ID: lil-512771

ABSTRACT

Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca²+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca²+]i oscillations similar to those evoked by sperm (95 vs 100 percent; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 µg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6 percent, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83 percent, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7 percent ethanol (62 vs 62 percent, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62 percent, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca²+]i oscillations, completion of meiosis and parthenogenetic development.


Subject(s)
Animals , Male , Mice , Calcium/metabolism , Culture Media, Conditioned/pharmacology , Cytochalasin B/pharmacology , Mesenchymal Stem Cells , Oocytes/drug effects , Parthenogenesis/drug effects , Microscopy, Confocal , Oocytes/physiology , Parthenogenesis/physiology
15.
Braz. j. med. biol. res ; 41(11): 978-985, Nov. 2008. ilus, graf
Article in English | LILACS | ID: lil-500368

ABSTRACT

Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), alpha-minimum essential medium (alpha-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), alpha-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with alpha-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.


Subject(s)
Animals , Female , Mice , Pregnancy , Culture Media, Conditioned/pharmacology , Mesenchymal Stem Cells , Oocytes/growth & development , Cumulus Cells/cytology , Embryo, Mammalian/embryology , Embryonic Development/physiology , Fertilization in Vitro , Meiosis/physiology , Ovarian Follicle/growth & development
16.
Braz. j. med. biol. res ; 41(5): 380-388, May 2008. ilus, tab
Article in English | LILACS | ID: lil-484437

ABSTRACT

Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a protein of 321 amino acids. Interestingly, homology analysis showed that the Der p 1 shared more than 87 percent identity in amino acid sequence with Eur m 1 but only 80 percent with Der f 1. Furthermore, phylogenetic analyses suggested that D. pteronyssinus was evolutionarily closer to Euroglyphus maynei than to D. farinae, even though D. pteronyssinus and D. farinae belong to the same Dermatophagoides genus. A total of three cysteine peptidase active sites were found in the predicted amino acid sequence, including 127-138 (QGGCGSCWAFSG), 267-277 (NYHAVNIVGYG) and 284-303 (YWIVRNSWDTTWGDSGYGYF). Moreover, secondary structure analysis revealed that Der f 1 contained an a helix (33.96 percent), an extended strand (17.13 percent), a ß turn (5.61 percent), and a random coil (43.30 percent). A simple three-dimensional model of this protein was constructed using a Swiss-model server. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Alignment and phylogenetic analysis suggests that D. pteronyssinus is evolutionarily more similar to E. maynei than to D. farinae.


Subject(s)
Animals , Allergens/immunology , Antigens, Dermatophagoides/genetics , Cloning, Molecular , DNA, Complementary/genetics , Escherichia coli/genetics , Mites/immunology , Amino Acid Sequence , Antigens, Dermatophagoides/isolation & purification , Blotting, Western , DNA, Complementary/chemistry , Dust , Molecular Sequence Data , Polymerase Chain Reaction , Recombinant Proteins/genetics , Sequence Analysis, DNA , Sequence Analysis, Protein
17.
Braz. j. med. biol. res ; 41(2): 110-116, Feb. 2008. ilus, tab
Article in English | LILACS | ID: lil-474763

ABSTRACT

To find Epstein-Barr virus (EBV) strains with genetic variations of EBV latent membrane protein 1 (EBV-LMP1) from nasopharyngeal carcinoma (NPC), the full-length DNA of LMP1 genes from 21 NPC biopsies obtained in Hunan province in southern China was amplified and sequenced. Our sequences were compared to those previously reported by the Clustal V method. Results showed that all 21 sequences displayed two amino acid changes most frequently in LMP1 of CD4+ T cell epitopes at codons 144 (F arrow right I, 21/21) and 212 (G arrow right S, 19/21) or (G arrow right N, 2/21). We also show that type A EBV strain is prevalent in the cases of NPC from Hunan province with a 30-bp 18/21 deletion, and we highlight that this deletion resulted in loss of one of the CD4+ T cell-restricted epitopes. The other 3 sequences without this deletion all had a change at codon 344 (G arrow right D). Furthermore, in the major epitope sequence of CD8+ T cells restricted by HLA-A2, all 21 sequences showed changes at codons 126 (L arrow right F) and 129 (M arrow right I). Our study discovered that one of the 21 sequence variations harbored a new change at codon 131 (W arrow right C), and 5/21 specimens showed another novel change at codon 115 (G arrow right A) in the major epitope sequence of CD8+ T cells restricted by HLA-A2. Our study suggests that these sequence variations of NPC-derived LMP1 may lead to a potential escape from host cell immune recognition, protecting latent EBV infection and causing an increase in tumorigenicity.


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Epitopes, T-Lymphocyte/genetics , Genetic Variation , /genetics , Nasopharyngeal Neoplasms/virology , Viral Matrix Proteins/genetics , Amino Acid Sequence , Biopsy , Epitopes, T-Lymphocyte/analysis , Genotype , Polymerase Chain Reaction , Sequence Analysis, DNA
18.
Southeast Asian J Trop Med Public Health ; 2006 May; 37(3): 452-5
Article in English | IMSEAR | ID: sea-35159

ABSTRACT

Adult mites' bodies of Dermatophagoides farinae were used as antigen in an indirect fluorescent antibody test (IFAT) to detect mite-specific IgG in sera of 48 patients with intestinal acariasis based on stool examination. Antibody titers with positive reaction ranged from 1:4 to 1:512 in 48 patients with intestinal acariasis. If antibody titers > or = 1:16 is regarded as being positive, the positive rate of patients detected with IFAT was 92%.


Subject(s)
Animals , Case-Control Studies , Feces/parasitology , Female , Fluorescent Antibody Technique, Indirect/methods , Humans , Intestinal Diseases, Parasitic/diagnosis , Male , Mite Infestations/diagnosis , Mites/classification
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