Objective To study the expression of signal transducers and activators of transcription 3(STAT3) in airway epithelial cells of asthmatic mouse model and its association with airway remodeling,explore the role of signal-transduction pathway in airway remodeling.Methods Thirty Balb/c mice were randomly divided into normal control group(n=10),asthma without intervention group(n=10) and AG490 intervention group(n=10).The mice were sensitized with ovalbumin to establish the asthmatic model.The histological changes were evaluated by HE staining,total brochial wall thickness(Wat)and smooth muscle thickness(Wam) were measured by image analysis system,the percentages of collagen deposition were detected by Masson′s trichrome staining,the expression of STAT3 in airway were detected by immunohistochemistry technique;lung tissue extracts were analyzed for phosphorylation of STAT3(p-STAT3)by Western blot.SPSS 13.0 software was used to analyze the data.Results 1.The histological changes including airway thickness,airway smooth cell proliferation and excessive collagen deposition in subepithelial aera were found under light microscope in asthmatic mice.2.The level of STAT3 and p-STAT3 in asthma without intervention group were significantly higher than those in normal control group(Pa

Objective To observe the effects of inhaled budesonide (BUD) in early phase on the airway inflammation in asthmatic mouse,and its effects on the IL-6/signal transducers and activators of transcription 3(IL-6/STAT3 )signaling pathway in airway,explore the therapeutic target of BUD.Methods Forty Balb/c mice were randomly divided into control group(n=10),asthma group(n=10),BUD group(n=10) and AG490 group(n=10).The mice were sensitized with ovalbumin to establish the asthmatic model.The histological changes were evaluated by HE staining.The levels of IL-4,IL-5 and IL-6 in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay.Lung tissue extracts were analyzed for total STAT3 and phospho-STAT3(p-STAT3) by Western blot.Results 1.The levels of inflammatory cells,EOS%, IL-4,IL-5 and IL-6 levels in the BALF in BUD group were significantly lower than those in asthma group (Pa0.05).Conclusions Inhaled corticosteroid can apparently ameliorate airway inflammation in early phase in asthmatic mouse model,and it can downregulate the expressions of IL-6 and STAT3,inhibit the signal-transduction pathway of STAT3.The IL-6 /STAT3 signaling pathway of airway may be one of the potential therapeutic targets of inhaled corticosteroid.

AIM: To purify human yolk sac mesenchymal ste m cells (hYS-MSC) and investigate its osteogenic and neurogenic differentiation potentials. METHODS: hYS-MSC were separated from yolk sac and purified via p assage culture. The karyotype of hYS-MSCs was analyzed via G-banded characterist ics. Flow cytometric analysis was used to determine the cell cycle and phenotype of hYS-MSC. The AKP expression of hYS-MSC was also tested. Osteogenic different iation of hYS-MSCs was induced by 10 -8mol/L dexamethasone, 10 mmol/L ?-gl ycerophosphate and 50 mg/L vitamin C. Alizarin red S stain was used for identifi cation of mineralization. ?-mecaptoethanol or salviae miltiorrhizae were used t o induce neurogenic differentiation of hYS-MSCs. The expressions of NSE, NF and GFAP were identified by immunohistochemical method. RESULTS: hYS-MSCs could be purified at passages 2 or 3. The cell cycle analysis suggested that hYS-MSCs showed strong proliferational potentials by which the cells kept normal diploid karyotype during the in vitro cultur e. Flow cytometry showed the phenotype of purified hYS-MSCs was uniformly positi ve for CD29, CD44, CD105, and CD166, and negative for reactivity to antigens CD3 4, CD45, or CD86. hYS-MSCs were weakly but clearly positive in AKP. Osteogenic d ifferentiation was appeared after induction of osteogenic differentiation. hYS-M SCs, which were of spindle shape, uniform in size, were induced to pleomorphism osteoblast-like cells which expressed high level of AKP. Aggregates or nodules w ere formed at day 7 and calcium accumulation was detected by alizarin red S stai ning on day 10 or day 14. Neurogenic differentiation of hYS-MSCs was induced by ?-mecaptoethanol or salviae miltiorrhizae. NSE, NF or GFAP positive cells were detected by immunohistochemical staining. CONCLUSIONS: hYS-MSCs have strong proliferation potential and th e normal diploid karyotype is kept during the in vitro culture. The phenoty pe of hYS-MSCs is coincident with adult hMSCs. hYS-MSCs could be induced to dif ferent iate into osteogenic or neurogenic cells.

OBJECTIVEEmbryonic stem cells (ESCs) are derived from totipotent cells of early embryo and they are potential to differentiate to any kind of cells of tissues in the body. Some reports showed that ESCs had broad capabilities of differentiating to variety of hematopotietic cells, such as erythroid, granulocyte/macrophage, megakaryocyte, mast and lymphocyte precursors. However, it is very difficult to control the phase of differentiation for ESCs in vitro. There is few report about hematopotietic stem cells (HSCs) from ESCs. Therefore, this research was designed to establish a culture system for generation of CD(34)(+)/Sca-1(+) HSC from ESC in vitro.

METHODSSingle mouse E 14.1 cells were suspended in methylcellulose medium, containing 40 ng/ml stem cell factor (SCF) and 20 ng/ml vascular endothelial growth factor (VEGF) and incubated at 37 degrees C with 5% CO2. In order to ensure the viability of the primary differentiation cultures over an extended period of time, the cultures were fed on day 7 with a dilute methylcellulose medium containing VEGF, SCF, interleukin-3 (IL-3), IL-6 and erythropoietin (EPO), which promoted their primary differentiation into embryoid bodies (EBs) with more CD(34)(+)/Sca-1(+) cells. Then, EBs with peak level of CD(34)(+)/Sca-1(+) cells were dispersed into single cells and replanted either in methylcellulose medium or in bone marrow stromal cells differentiation system containing 15% fetal bovine serum (FBS), 160 ng/ml SCF, 20 ng/ml VEGF, 30 ng/ml IL-3, 30 ng/ml IL-6, 3 U/ml EPO and 20% BIT for HSC into second-step differentiation. The HSCs were characterized by flow cytometric analysis, colonogenic cell assay and Wright-Giemsa stains.

RESULTSVEGF had the strongest stimulatory effect on the enhancement of the CD(34)(+)/Sca-1(+) cells population when combined with SCF, IL-3, IL-6 and EPO. It could markedly accelerate mouse E14.1 cells to differentiate into EB with more CD(34)(+)/Sca-1(+) cells. Cell cytometric analysis showed CD(34)(+)/Sca-1(+) cells were up to (1.91 +/- 0.40)% by day 5 and (8.11 +/- 1.17)% by day 8, and the peak level of CD(34)(+)/Sca-1(+) cells was (13.72 +/- 1.92)% by day 12. However, CD(34)(+)/Sca-1(+) cells could not increase in number with the prolongation of differentiation. So renewal single cells suspension from EB by day 12 was dispersed into the second step differentiation. The results showed that HSC was slowly generated with a few hematopoietic colony formations in methylcellulose medium differentiation system. CD(34)(+)/Sca-1(+) cells got (2.74 +/- 0.80)% by day 5 and (11.37 +/- 1.84)% by day 10, and apex percentage of CD(34)(+)/Sca-1(+) cells was about (20.52 +/- 2.78)% by day 14. However, EBs generated quickly for HSC with increased hematopoietic cell population by co-culture on bone marrow stromal cells feeder. Flow cytometric analysis showed that the percentages of CD(34)(+)/Sca-1(+) cells was (7.33 +/- 1.61)% by day 5, (13.28 +/- 2.59)% by day 8, and (20.81 +/- 3.19)% by day 10. EB cells were induced after 12 days to reach the peak level of (34.60 +/- 3.71)%. Hematopoietic colony formation unit (CFU) analysis showed that CFU was sufficient from cells on bone marrow stromal cells differentiation system in the second step compared to that in methylcellulose medium differentiation system, and Wright-Giemsa stain could confirm its characteristics of hematopoietic progenitors.

CONCLUSIONUsing two-step differentiation, the investigators got a good way to control the phase of differentiation from ESC to HSC. The bone marrow stromal cell differentiation system combining with VEGF, SCF, IL-3, IL-6 and EPO was an optimal system for the generation of HSC with CD(34)(+)/Sca-1(+) surface marker from ESC differentiated in vitro. This study demonstrated that these cells could form more hemopoietic colonies.

Animals ; Antigens, CD34 ; Cell Culture Techniques ; Cell Differentiation ; drug effects ; Cell Survival ; Cells, Cultured ; Embryonic Stem Cells ; drug effects ; immunology ; Hematopoietic Stem Cells ; physiology ; Mice

UNLABELLEDProminent eosinophil airway inflammation is important in the pathogenesis of asthma. There is increasing evidence that the disorder of eosinophil apoptosis contributes to the mechanism. But most of the studies have been done in vitro or on animal models, very few were done among the adult asthmatics in vivo.

OBJECTIVEThe aim of this study was to elucidate the relationship between the apoptotic eosinophils and Bcl-2 in asthmatic children in vivo.

METHODSEleven mild to moderate asthmatic patients were recruited and the range of age was 7 - 14 years (9 males, 2 females), meanwhile 7 patients with lower respiratory infection were recruited as control and the range of age was 9 - 14 years (5 males, 2 females). Before and after inhaled glucocorticoid (GC) induced sputum, bronchoalveolar lavage (BAL), bronchial mucosa specimens and peripheral blood were obtained for measuring and comparing the changes of apoptotic EG(2)(+) cell by combining the techniques of TUNEL and immunohistochemistry, meanwhile the expression of Bcl-2 in bronchial mucosa specimens was measured by using the immunohistochemical assay.

RESULTSBefore the inhalation of GC, the apoptotic EG(2)(+) cells in asthmatics were significantly lower than that in control group (P < 0.01), and the numbers of EG(2)(+) cell in asthmatics group were significantly higher than that in control group (P < 0.001). After the treatment apoptotic EG(2)(+) cells in asthmatics were increased (P < 0.01), and the numbers of EG(2)(+)cell were decreased (P < 0.01, P < 0.05 and P < 0.05, respectively), FEV(1)% was increased (P < 0.05). Before the inhalation of GC, the numbers of Bcl-2(+) cell in asthmatic airway submucosa were higher than that in control group (P < 0.05) but after the treatment the number of Bcl-2(+) cell did not change significantly. (4) Before and after GC treatment the percentages of apoptotic eosinophils of peripheral blood in vivo had no significant changes compared with those of control subjects (P > 0.05). There was a positive correlation between apoptosis of EG(2)(+) cell in sputum, BAL, airway submucosa and FEV(1)% (P < 0.05).

CONCLUSIONApoptosis of EG(2)(+) cell decreased in the airway of asthmatic children and inducing EOS apoptosis is one of the important mechanism of inhaled GC therapy for asthma.

Adolescent ; Apoptosis ; Asthma ; blood ; drug therapy ; pathology ; Bronchoalveolar Lavage Fluid ; cytology ; Child ; Eosinophils ; cytology ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Respiratory Mucosa ; chemistry ; cytology

OBJECTIVETo investigate the association between Arg16Gly polymorphism of beta(2)-adrenergic receptor (ADRB2) gene and blood pressure levels.

METHODSA total of 487 hypertensive individuals were recruited from YueXi county of Anhui province. 672 patients' parents and siblings were also invited to take part in the study and used as genomic control. Blood pressure was measured and a standardized questionnaire regarding social-economic characteristics, general health status, occupational history and life-style and dietary factors was administered for each participant. The ADRB2 Arg16Gly polymorphism was genotyped by using a PCR-Restriction Fragment Length Polymorphism (RFLP) method. The association between the ADRB2 polymorphism and hypertension in hypertensive adults was determined by utilizing a family-based association test analysis.

RESULTSIn this study population, carriers of the ADRB2 Arg16 allele had lower systolic (P < 0.01) and diastolic (P < 0.01) blood pressure, suggesting that the genetic effect on blood pressure was more likely to fit an additive model.

CONCLUSIONOur results suggest a probable association between Arg16Gly polymorphism of ADRB2 gene and hypertension.

Adult ; Asian Continental Ancestry Group ; genetics ; Blood Pressure ; China ; epidemiology ; Female ; Genotype ; Humans ; Hypertension ; epidemiology ; genetics ; Male ; Middle Aged ; Pedigree ; Polymorphism, Single Nucleotide ; Receptors, Adrenergic, beta-2 ; genetics ; Young Adult

OBJECTIVEExtraovarian peritoneal serous papillary carcinoma (EPSPC) is both histologically and clinically similar to stage III-IV ovarian papillary serous carcinoma (OPSC). The purpose of this study is to investigate the clinical findings, treatment, and outcome of EPSPC patients compared with stage III-IV OPSC patients.

METHODSThe data of 12 EPSPC patients and 45 stage III-IV OPSC patients were retrospectively reviewed, comparing the characteristics on clinical presentation and treatment, sensitivity to first-line chemotherapy agents and survival.

RESULTSBy analysis of patients' characteristics, presenting signs and symptoms, type and extent of surgery, tumor response to first-line chemotherapy, recurrence-free interval, recurrence site and serum CA-125 levels, no significant difference was observed between the EPSPC patients and stage III-IV OPSC controls. The prevailing presenting symptoms were abdominal mass and ascites. The mainstay of treatment was debulking surgery followed by adjuvant platinum-based chemotherapy. The complete clinical response of stage III-IV OPSC was 91.8% compared with 25.0% for women with EPSPC (P < 0.01).

CONCLUSIONThe clinical and surgical characteristics of EPSPC are similar to those of stage III-IV OPSC. When the same treatment strategy is applied, similar response and survival are expected in either condition.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CA-125 Antigen ; blood ; Cisplatin ; therapeutic use ; Combined Modality Therapy ; Cyclophosphamide ; therapeutic use ; Cystadenocarcinoma, Papillary ; blood ; drug therapy ; pathology ; surgery ; Disease-Free Survival ; Doxorubicin ; analogs & derivatives ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Neoplasm Staging ; Ovarian Neoplasms ; blood ; drug therapy ; pathology ; surgery ; Paclitaxel ; Peritoneal Neoplasms ; blood ; drug therapy ; pathology ; surgery ; Retrospective Studies ; Taxoids ; therapeutic use

OBJECTIVETo investigate the effect of nasal cavity expansion surgery on the abnormal blood supply of the cerebral arterial system.

METHODSFifty-nine inpatients with abnormal blood supply of cerebral arterial system confirmed by transcranial doppler (TCD) and chronic nasal obstructive diseases were included in this study. All patients accepted nasal cavity expansion surgery and were followed-up with TCD every month after operation until TCD became normal, or up to seven months even if the TCD was still abnormal. SPSS 17.0 software was used to analyze the data.

RESULTSIn all 59 patients, there were 164 TCD-abnormal cerebral arteries. Among them, 37 patients(62.71%) with abnormal TCD arteries became normal within 1 to 7 months after operation, 8 patients (13.56 %) got better, but 14 patients (23.73 %) did not improve.

CONCLUSIONSAbnormal blood flow of some cerebral arteries was possibly induced by increasing the activation of sympathetic nervous system around the vertebral arterial system, caused by chronic nasal obstruction. Nasal dilatancy surgery can improve the blood supplement of the cerebral arterial system.

Blood Flow Velocity ; Cerebral Arteries ; Cerebrovascular Circulation ; Humans ; Nasal Cavity ; Ultrasonography, Doppler, Transcranial

The study was aimed at the exploration of relationship between T cells expressing killer cell inhibitor receptors (KIR, CD158 and CD94) and graft-versus-host disease (GVHD) after hematopoietic stem cell transplantation. The expression rates of CD158a, CD158b and CD94 on T cells and NK cell were detected by flow cytometry and donor/recipient HLA-Cw was analyzed using PCR after peripheral blood stem cell transplantation (PBSCT) and umbilical cord blood transplantation (UCBT). After both PBSCT and UCBT, the rates of CD3(+)CD158a(+) and CD3(+)CD158b(+) T cells increased, especially the rate of CD8(+)CD158b(+) T cells. In both acute and chronic GVHD groups, the rate of CD3(+)CD158b(+) T cells increased, especially in acute GVHD. The CD94 mainly expressed on CD3(+)CD8(+) T cells. The percentage of the expression of CD94 on CD4(+) and CD8(+) cells after UCBT and PBSCT increased significantly. The expression of KIR in GVHD (early stage of transplantation) increased but the expression of KIR in chronic GVHD (advanced stage of transplantation) decreased. Five patients who HLA-Cw matched had no severe GVHD. In four patients who underwent allo-PBSCT and UCBT from related HLA-matched donors, only 2 patients had no aGVHD. Four patients underwent transplantation from unrelated HLA-matched donors had GVHD. These observations suggested that there is some relationship between GVHD and KIR expression on T cells. CD158b might be an inhibitory molecule of T cell activated at early stage after transplantation. Understanding the mechanism of GVHD with the expression of KIR on T cells, especially those binding the HLA-Cw might shed light on the establishment of the specific immunotolerance for the prevention of GVHD. To pay attention to HLA-Cw typing is very important to reduce GVHD and increase GVL effect in related or unrelated HLA-matched transplantation.
Antigens, CD ; analysis ; Antigens, Differentiation, T-Lymphocyte ; analysis ; Genotype ; Graft vs Host Disease ; etiology ; HLA-C Antigens ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Lectins, C-Type ; Receptors, Immunologic ; analysis ; Receptors, Interleukin-2 ; analysis ; Receptors, KIR ; Receptors, KIR2DL1 ; Receptors, KIR2DL3 ; T-Lymphocytes ; immunology

To investigate effects of rat bone marrow mesenchymal stem cells (rBMMSC) on hematopoiesis after allo-hematopoietic stem cell transplantation (HSCT), allogeneic BMT model from Fischer 344 rats (RT-1Al) to Wistar rats (RT-1Au) was established; effects of MSCs on hematopoietic reconstitution were studied by survival rate, peripheral blood counts, histological analysis and FACS at day 30 after transplantation. The results showed that (1) MSCs from donor Fisher344 could survive in recipient irradiated by lethal dose and could be found in the thymus, spleen and bone marrow of the recipient at 30 days after cotransplantation with BM by measuring EGFP gene. (2) Cotransplanation of MSCs and BM improved hematopoietic reconstitution. Lymphocyte and platelet counts of peripheral blood in cotransplantation group were higher than those in the control group. Active hematopoiesis and increase of bone marrow nucleated cells were observed in cotransplantation group. MSCs significantly enhanced hematopoiesis of B lymphocyte and megakaryocytopoietic lineages by FACS analysis. It is concluded that (1) MSCs of Fisher344 can be found in the thymus, spleen, bone marrow of the recipients at 30 days after cotransplantion by measuring EGFP gene. (2) hematopoietic reconstitution is significantly enhanced by MSCs cotransplanted with BM.
Animals ; Bone Marrow Transplantation ; methods ; Cell Differentiation ; physiology ; Flow Cytometry ; Hematopoiesis ; physiology ; Lymphocyte Count ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; physiology ; Models, Animal ; Platelet Count ; Rats ; Rats, Inbred F344 ; Rats, Wistar