RESUMO
OBJECTIVE@#Chronic rhinosinusitis (CRS) involves inflammation of the nasal and para-nasal mucosa. Due to its heterogeneous nature, unknown pathogenesis, and high recurrence rate, effective treatment is difficult. Nasal cytology is presently not a part of the routine diagnosis or treatment decision for CRS.@*DATA SOURCES@#A literature search was performed for published papers in English between January 1990 and June 2019 using MEDLINE.@*STUDY SELECTION@#Terms used were chronic rhinosinusitis, eosinophils, etiology, immunopathology, inflammation, mast cells, nasal cytology, polyps, and treatment. Both reviews and original articles were collected and studied.@*RESULTS@#There is no standard nasal fluid, mucus sampling, or staining techniques for identifying inflammatory cell types. Results were divergent from different countries. Moreover, the main focus of these papers on the cells in nasal washings was eosinophils, with infrequent mentioning of other cell types that may imply different etiology and pathology. The heterogeneous cell profile of CRS and the role of mast cells have been unappreciated due to the lack of specific immunohistochemical technique or study of its unique mediators.@*CONCLUSIONS@#Nasal cytology could help distinguish the type and the activation state of inflammatory cells. Thus it can help in providing a clearer picture of CRS pathogenesis, identifying different patient groups, and developing effective treatments.
RESUMO
Objective@#Hepatocellular carcinoma (HCC) is one of the most common malignant tumor worldwide. Metastasis is a marker of cancer deterioration in patients with liver cancer and a major cause of death. In order to develop effective therapeutic strategies, it is urgent to study the molecular basis of liver cancer metastasis.@*Methods@#Immunohistochemistry was used to detect the expression of fatty acid synthase (FASN) in HCC. Wound healing and transwell cell invasion assays was used to confirm the role of FASN in liver cancer migration and invasion. Proteins that interacted with FASN were identified using iTRAQ (isobaric tag for relative and absolute quantification). Co-immunoprecipitation (Co-IP) and cellular immunofluorescence analysis were used to assess the interaction between FASN and signal transduction and transcription activator 3 (STAT3). The expression of STAT3, p-STAT3, matrix metalloproteinase (MMP)-2 and MMP-9 was detected after FASN knockdown using Western blot method. Statistical analysis was performed using the t-test.@*Results@#Immunohistochemistry showed that the expression of FASN in HCC tissue was higher than that in adjacent tissues. iTRAQ, Co-IP and immunofluorescence analysis revealed that FASN interacted with STAT3. Western blot analysis showed that the expression of p-STAT3, MMP-2 and MMP-9 decreased after FASN knockdown.@*Conclusion@#FASN may promote the metastasis of liver cancer by interacting with STAT3 and affecting the expression of MMP-2/MMP-9.
RESUMO
Objective@#Chronic rhinosinusitis (CRS) involves inflammation of the nasal and para-nasal mucosa. Due to its heterogeneous nature, unknown pathogenesis, and high recurrence rate, effective treatment is difficult. Nasal cytology is presently not a part of the routine diagnosis or treatment decision for CRS.@*Data sources@#A literature search was performed for published papers in English between January 1990 and June 2019 using MEDLINE.@*Study selection@#Terms used were chronic rhinosinusitis, eosinophils, etiology, immunopathology, inflammation, mast cells, nasal cytology, polyps, and treatment. Both reviews and original articles were collected and studied.@*Results@#There is no standard nasal fluid, mucus sampling, or staining techniques for identifying inflammatory cell types. Results were divergent from different countries. Moreover, the main focus of these papers on the cells in nasal washings was eosinophils, with infrequent mentioning of other cell types that may imply different etiology and pathology. The heterogeneous cell profile of CRS and the role of mast cells have been unappreciated due to the lack of specific immunohistochemical technique or study of its unique mediators.@*Conclusions@#Nasal cytology could help distinguish the type and the activation state of inflammatory cells. Thus it can help in providing a clearer picture of CRS pathogenesis, identifying different patient groups, and developing effective treatments.
RESUMO
Objective To determine the susceptibility genes and resistance genes in HLA-DRB1 alleles in Tibetan patients with cystic and alveolar hydatid diseases,so as to provide the references for the research of the genetic characteristics and infec-tion mechanism of Tibetan hydatid diseases. Methods The case control method was applied. The Tibetan patients with cystic and alveolar hydatid diseases(63 and 73 cases respectively)in Yushu and Guoluo Tibetan Autonomous Prefecture,and unrelat-ed healthy people(60 cases)in this area were selected as the study subjects. The polymerase chain reaction-sequence based typ-ing(PCR-SBT)technique was applied for genotyping of HLA-DRB1,and the comparison of the gene frequency. Results The frequency of HLA-DRB1*04 in the alveolar/cystic echinococcosis group was lower than that in the control group(χ2 =4.71, 4.31,both P<0.05). Conclusion HLA-DRB1*04 genotypes may be associated with the resistance of cystic and alveolar echi-nococcosis and its resistance genes.
RESUMO
This study was purposed to construct and prepare the recombinant adenovirus vector carrying human thioredoxin (hTRX) and enhanced green fluorescence protein (EGFP), and transfect it into HEK293 cells, so as to lay a foundation for further gene therapy. The PCR-amplified products of hTRX with a pair of primers containing Not I and EcoR V restriction sites were subcloned into shuttle plasmid pDC316-mCMV. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid pDC316-hTRX-EGFP and large adenovirus-helper plasmid pBHGlox (delta) E1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus pAd-hTRX-EGFP was co-transfected in HEK293 cells, purified by CsCl gradient centrifugation, counted for virus particles and determined for titer. The recombinant adenovirus was identified by PCR. The HEK293 cells were then transfected with adenoviruses and assayed by flow cytometry. The expression of hTRX was confirmed by Western blot. The results showed that according to PCR and restriction endonuclease assay, the target gene was inserted into recombinant adenovirus vector successfully. The sequence of fusion gene was the same as that of designed fragments. The titer of the purified recombinant adenovirus pAd-hTRX-EGFP was 5.558×10(10) pfu/ml. A transfection efficiency of 92.25% could be achieved at MOI = 100. Western blot further confirmed that hTRX was efficiently expressed in HEK293 cells. It is concluded that recombinant adenovirus vector containing hTRX has been constructed successfully and obtained highly efficient virus that can express efficiently in HEK293 cells, which laid a foundation for further investigation.
Assuntos
Humanos , Adenoviridae , Genética , Vetores Genéticos , Proteínas de Fluorescência Verde , Genética , Células HEK293 , Plasmídeos , Proteínas Recombinantes , Genética , Tiorredoxinas , Genética , TransfecçãoRESUMO
Objective To study the expression levels ofinterlenkin-6 (IL-6), IL-6 receptor (IL-6R),its soluble IL-6 receptor (sIL-6R) and soluble glycoprotein 130 (sgp130) in brain tissues of refractory temporal lobe epilepsy (TLE) and explore their role in TLE. Methods Forty patients with TLE and other 40 patients with trauma/hypertensive cerebral hemorrhage (controls), admitted to and performed surgery in our hospital from January 2010 to December 2010,were chosen in our study.Brain tissues from temporal lobe were collected during the surgery.By using real time-PCR and ELISA,the mRNA or protein expression levels of IL-6,IL-6R,sIL-6R and sgp130 were analyzed. Results The mRNA expressions of IL-6 and IL-6R in the TLE brain tissues were significantly higher as compared with those in controls (P<0.05).ELISA indicated that sIL-6R level in the TLE brain tissues was significantly higher than those in controls (P<0.05); however,sgp130 level in the TLE brain tissues was slightly higher than those in controls without significant difference (P>0.05). Conclusion The high level of IL-6 may influence the function of varies types of cells within the epileptic foci of patients with TLE through classical signaling pathway or trans-signaling pathway, which is potentially involved in the epileptogenesis of TLE.
RESUMO
<p><b>OBJECTIVE</b>To study the possibility and mechanism of construction of tissue engineered bone with human bone marrow mesenchymal stem cells (hBMSCs) as seeding cells and partially demineralized bone matrix (pDBM) as scaffold.</p><p><b>METHODS</b>hBMSCs are cultured and mutiplified. The 4th grade hBMSCs are seeded on the pDBM, the growth and adhesion of hBMSCs on pDBM are observed under scanning electro microscope. The adhesion efficiency is assessed. The complexes are implanted in the nude mice subcutaneously, the pDBM without cells as control. The grafts are taken out on the 8th and 12th week.</p><p><b>RESULTS</b>There is new bone formation on the 8th and 12th week in complex group. There is a layer of osteoblast like cells adhered on the surface of most of the new bone, which suggest the possibility of intramembranous ossification. There is no bone formation in control group.</p><p><b>CONCLUSIONS</b>Tissue engineered bone can be constructed with hBMScs and pDBM in vivo, and the mechanism of which could be intramembranous ossification.</p>
Assuntos
Animais , Humanos , Camundongos , Células da Medula Óssea , Biologia Celular , Osso e Ossos , Adesão Celular , Diferenciação Celular , Células-Tronco Mesenquimais , Biologia Celular , Camundongos Nus , Engenharia Tecidual , MétodosRESUMO
Objective To investigate the effects of cryopreservation on the growth and osteogenesis capa- bility of human bone marrow stromal cells(BMSCs)on demineralized bene matrix(DBM).Methods Bone marrow aspirates were obtained from the lilac crests of three donors.The BMSCs were isolated from the bone marrow by density gradient centrifugation.Cells of passage 3 were cryopreserved in liquid nitrogen for 24 hours,and then re- covered.The non-cryopreserved BMSCs were used as the control,The cryopreserved and control BMSCs were cul- tured in osteogenic media,collected and labeled with Dil to be seeded onto the DBM when cells were confluent.The percentage of BMSCs adhered to the DMB was detected.The cell morphology and matrices secreted by BMSCs on the DBM were observed by the inverted phase-contrasted microscope,fluorescence microscope and scanning electron microscope(SEM).The growth and viability of BMSCs on the DBM were determined using the modified MTT ashy. The osteogenesis ability of BMSCs on the DBM was determined by assessment of the alkaline phosphatase(ALP) activity and osteocalcin(OCN)content.Results The percentages of the cryopreserved and control cells adhered to DBM were(97.25?1.17)% and(97.00?1.09)% respectively.The cells adhered well to the DBM and grew rapidly.Large amounts of matrices on the DBM were observed by the light microscope and SEM.The cells embedded in the matrices could be observed by fluorescence microscope.There were no significant differences in the assay values of MTT,ALP and OCN between the cryopreserved and control BMSCs on the DBM.Conclusion Since cryopreservation does not affect the growth and osteogenesis capability of BMSCs on DBM,the cryopreserved BMSCs can be used as a cell source in bone tissue engineering.