After human umbilical vein endothelial cells (HUVECs) were incubated with various concentrations of exenatide for 24,48,and 72 h under the conditions of5.5 and 33 mmol/L glucose,the cell viability was detected by MTF assay.The apoptosis rate of endothelial cells was measured by flow cytometry.Protein kinase B (Akt) phosphorylation,Bcl-2,and Bax protein expression were detected by Western blotting.The results showed that the cell viability was decreased after HUVECs were incubated with high glucose for48 and 72 h (P＜0.01).Treatment with 1,10,and 100 nmol/L exenatide for48 h significantly increased the cell viability in the presence of high glucose,in a dose-dependent manner (P＜0.01).High glucose inhibited Akt phosphorylation and Bcl-2 protein expression,stimulated Bax protein expression,with decreased Bcl-2/Bax ratio and increased apoptosis ratio of cells.After treatment with exenatide,the cell apoptosis reduced,Akt phosphorylation and Bcl-2 protein expression increased,and Bax protein expression decreased,with increased Bcl-2/Bax ratio (P ＜ 0.01).These effects of exenatide on endothelial cell were abrogated by an inhibitor of phosphotylinosital 3 kinase (PI3K) LY294002 (P ＜ 0.01),suggesting that the exenatide may regulate Bcl-2/Bax protein expression and inhibit endothelial cell apoptosis induced by high glucose through PI3k/Akt signal pathway,and thus may protect endothelial cells.

Gene engineering is the main course of biological engineering. It should be adapted to the demand of innovation spirit, practice ability and comprehensive quality of students. Educational reform of gene engineering conducted by constructing system of theory and practice, optimizing course teaching content, strengthening practice teaching content, using modern teaching technology, strengthening web course construction and improving teaching methods. We pay attention to impart specialty knowledge and learning methods to students. Its aim was to increase teaching effects and meet the demands of bioengineering specialty and qualified personal training in 21 century.

Objective To observe the expressions of matrix metalloproteinase-9 mRNA and the change of cerebral edema after diffuse brain injury in rats and discuss their correlation.Methods Marmaruu's diffuse brain injury model of rat was made.Real-time quantitative RT-PCR,dry-wet meth- od,histological techniques and electron microscope were used to determine the expressions of MMP-9 containing water in brain tissue and inflammatory reaction and uhrastructural changes of blood capillary at different time phases after truama.Results The expressions of MMP-9 mRNA started to increase at 1 hour,peaked at 12 hours(P

OBJECTIVETo study the clinical characteristics of Streptococcus pneumonia-associated hemolytic uremic syndrome (SP-HUS) in children.

METHODClinical and laboratory data of a pediatric case of SP-HUS were retrospectively analyzed and the key points of diagnosis and therapy were reviewed.

RESULTAn 18-month old girl was admitted with chief complaint of fever and cough for 5 days combined with mild labored breath. Breath sound was found weakened in right lung with lower lobe dullness on percussion. Laboratory tests revealed: WBC 3.7×10(9)/L, Hb 83 g/L, PLT 11×10(9)/L, C-reactive protein (CRP) > 180 mg/L. Morphological study of the RBCs showed marked anisocytosis and schistocytosis. Urinalysis showed 42.66 RBCs per high-power field, occult blood (+++), proteinura (++++). Streptococcus pneumoniae was isolated from blood, pleural fluid and sputum. Serotyping with simplified chessboard system was 3. The direct Coombs test was positive. Serum complement levels (C3 and C4) were depressed at 0.699 g/L, 0.064 g/L, respectively. Chest X-ray showed pleural effusion and infection of the right hemothorax. The computerized tomographic scan of the chest revealed pneumatoceles in the right lower lobe. The diagnosis on admission we considered was SP-HUS. Intravenous antibiotic therapy (vancomycin + cefoperazone/sulbactam) was administered. The renal replacement theraphy was administered to maintain electrolyte and fluid balances and adequate nutrition. Transfusions of washed red blood cells were administered to correct the anemia. One month after admission the patient was good with recovery. Liver and renal function recovered and the pneumonia was resolving, anemia and platelets were corrected. The direct Coombs test turned to be negative. Serum complement levels (C3 and C4) were normal. After 3-month follow-up, no clinical anomalies were detected.

CONCLUSIONSP-HUS should be suspected when the following occurs in the context of pneumococcal infections: microangiopathic hemolytic anemia, thrombocytopenia, acute renal failure and a positive Coombs test result. Serotype 3 of SP was associated with HUS.

Anti-Bacterial Agents ; therapeutic use ; Biomarkers ; analysis ; Coombs Test ; Female ; Hemolytic-Uremic Syndrome ; diagnosis ; etiology ; microbiology ; therapy ; Humans ; Infant ; Lung ; diagnostic imaging ; pathology ; Pleural Effusion ; etiology ; Pneumococcal Infections ; complications ; Radiography ; Retrospective Studies ; Serotyping ; Streptococcus pneumoniae ; classification ; isolation & purification

The purpose of this study was to evaluate the safety of cryopreserved and thawed peripheral blood stem cell (PBSC) fractionated return infusions in children. 35 children patients with malignant tumors (13 acute leukaemias, 15 neuroblastomas and 7 malignant lymphomas) received fractionated return infusions of cryopreserved stem cells after undergoing high-dose chemotherapy without or with total body irradiation. The toxicities of 70 return infusions were evaluated. All patients were mobilized by chemotherapy plus recombination human granulocyte colony-stimulating factor (rhG-CSF), and then PBSCs were collected by a separator CS-3000 plus or COBE spectra-4. The grafts were cryopreserved in 10% dimethyl sulfoxide (DMSD) and stored in liquid nitrogen. There were totally 70 PBSC transfusions. The total volume of PBSCs transfused: 190 - 420 ml (265 +/- 73 ml or 13.7 +/- 4.2 ml/kg) with a mean of (4.43 +/- 1.91) x 10(8)/kg of PBSCs, and 0.94 +/- 0.18 g/kg of DMSO. The single dose: 90 - 300 ml (132 +/- 37 ml or 6.6 +/- 5.2 ml/kg) with a mean of 0.68 +/- 0.12 g/kg of DMSO. Symptoms occurring during the infusions were recorded. All patients were monitored for 24 hours after infusion. Pulse, blood pressure, body temperature, and respiratory rate were recorded every 15 minutes. At four hours before and 8 hours after infusion, urinalysis was performed. Serum potassium, sodium, creatinine, total bilirubin, aspartate amino transferase (AST), and alanine amino transferase (ALT) levels were examined within 24 hours before and after the first infusion. The results showed that the toxicities observed included hemoglobinuria in 54 return infusions (77.1%), headache in 28 (40.0%), nausea in 24 (34.3%), vomiting in 17 (24.3%), and abdominal pain in 8 (11.4%). Patients who received a graft > 200 ml tended to have a higher frequency of hemoglobinuria, headache, nausea, vomiting, or abdominal pain (P<0.01), and they disappeared quickly, too. Total bilirubin increased after the first return infusion (P<0.01), and there was a significant correlation between the volume of infusion and the degree of total bilirubin increase (r=0.8977, P<0.01). No renal failure or shock occurred. It is concluded that transient hemoglobinuria, headache, nausea, vomiting, and abdominal pain are common toxicities associated with PBSC autograft, and these toxicities are related with a single volume of PBSCs transfused. Total bilirubin increase is correlated with the volume of infusion. In a word, the toxicity is less frequent and lower severe in children with fractionated infusions of cryopreserved peripheral blood stem cell.
Acute Disease ; Adolescent ; Child ; Child, Preschool ; Cryopreservation ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Headache ; etiology ; Hematopoietic Stem Cell Mobilization ; methods ; Hemoglobinuria ; etiology ; Humans ; Leukemia ; therapy ; Lymphoma ; therapy ; Male ; Nausea ; etiology ; Neoplasms ; therapy ; Neuroblastoma ; therapy ; Peripheral Blood Stem Cell Transplantation ; adverse effects ; methods ; Recombinant Proteins

OBJECTIVETo investigate the characteristics of transurethral partial cystectomy with a 2 µm laser in diagnosis and treatment for the bladder submucosal lesions in adults.

METHODSNine patients with suspected pathological diagnosed bladder submucosal lesions in out-patient department were diagnosed and treated transurethral with a 2 µm laser under sacral block between August 2009 and December 2010. The diameters of tumors were 1.5 - 2.5 cm. A 2 µm laser was used to incise the full-thickness bladder wall around the tumors. The entire bladder wall was peeled between the detrusor muscle layer and outer connective tissues. Tumors with bladder wall at the base were removed together and sent for pathological examination. The surgical procedures, intraoperative hemorrhage, intraoperative and postoperative complications were observed, pathological diagnosis and postoperative follow-up were performed.

RESULTSAll operations were successful. Mean operative time was 36.4 minutes (range 25 to 47 minutes), perioperative blood loss was minimal. There was no obturator nerve reflection and no hemorrhage detected after surgery. Postoperative pathological diagnosis included leiomyoma in 3 cases, pheochromocytoma in 3 cases, endometriosis in 1 case and metastatic bladder cancer in 2 cases.

CONCLUSIONSTransurethral partial cystectomy with a 2 µm laser can diagnose and treat bladder submucosal lesions. The procedures are effective and safe. Patients could get accurate pathological diagnosis without further painful and some bladder tumors can be treated by minimally invasive surgery.

Adult ; Aged ; Cystectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Laser Therapy ; Male ; Middle Aged ; Treatment Outcome ; Urinary Bladder Neoplasms ; surgery

Objective To detect the feasibility of magnetically labeled swine bone marrow mesenehymal stem cells(MSCs)with SHU 555A combined with poly-L-arginine(PLL),under MR imaging in vitro and in vivo.Methods Swine mesenehymal stem cells were isolated and culture-expanded 3 passages in vitro,then magnetically labeled by incubation with SHU 555A(25?g Fe/ml,Resovist,Schering)for 24 hours with 750 ng/mL poly-L-lysine(PLL;average MW_275 kDa)added 1 hour before incubation.Cellular iron incorporation and detention at 0 d,4 d,8 d,12 d,16 d,20 d after labeling was qualitatively assessed using Prussian blue and quantified at atomic absorption spectrometry.Cell viability was assessed by trypan-blue exclusion test.Cell suspensions underwent MR imaging with T_1-and T_2-weighted spin-echo and fast field-echo sequences on a clinical 1.5 T MR system.At last,1?10~6 SHU 555A labeled and unlabeled MSCs were transextracardially implanted into the infracted and normal myocardium approximately 2 week following the ligation of left anterior descending coronary artery in 1 swine respectively,and finally performed 1.5-T MRI within 1 week after infarction.Results①Intracytoplasmic particles stained with Prussian blue stain were detected for all cells with mean cellular iron content of(13.13?2.30)pg per cell.With division of stem cells, the stained particles decreased gradually with iron content(0.68?0.20)pg per cell.at 16 days after labeling, approximately to the prelabeled baseline values.(0.21?0.06)pg per cell(P＞0.05).The viability of the labeled cells at various time points were not significantly different with that of nonlabeled cells(P＞0.05).②MR images showed signal intensity changed most obviouly in T2*WI in vitro.The percentage change of signal intensity increased with increasing cell numbers,and decreased with the time.As few as 5?10~4-1?10~5 cells could be detected by using this approach.③Two injected sites containing MR-MSCs were detected in vivo,presentingas low signal intensity areas with the T_2*WI scanning sequence.Conclusion Swine bone marrow MSCs can be labeled with SHU555A-PLL and depicted with a standard 1.5-T MR imager in vitro and in vivo.(J lntervent Radiol,2007,16:115-121)

Objective To investigate the molecular mechanisms of upregulated expression of cellular Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein(c-FLIP)by calreticulin(CRT)in patients with rheumatoid arthritis (RA). Methods The semi-quantitative analysis and localization of c-FLIP in RA and osteoarthritis (OA)synovium were detected by immunohistochemistry.The fibroblast-like synoviocytes(FLS)were isolated by enzymatic digestion of synovial tissue specimens obtained from RA and OA patients,and cultured as an in vitro experiment model.The expressions of c-FLIP in RA and OA synovial fibroblasts were detected by immunofluorescence and Western blot assay. Whether CRT influenced c-FLIP expression and its molecular mechanism were explored by Western blot assay. Results The high expression of c-FLIP was found in RA synovium, mainly in the lining and sublining areas of FLS and vascular endothelial cells detected by immunohistochemistry.Meanwhile,weak staining of c-FLIP was observed in OA synovium.The expression of c-FLIP was significantly higher in RA synovium than that of OA synovium(t=11.717,P<0.001).Results of immunofluorescence and Western blot assay showed that c-FLIP was mainly located in cytoplasm, and which was higher expressed in FLS of RA than that of OA. The increased c-FLIP expression and phosphorylation of NF-κB were detected after being co-incubated with exogenous CRT (0, 10, 50, 100 μg/L), in dose-dependent manner. The effect of CRT upregulating c-FLIP expression was blocked by NF-κB inhibitor BAY 11-7082.Conclusion CRT can increase c-FLIP expression at least partly through NF-κB pathway in RA,which may provide therapeutic target for the treatment of RA.

Objective To determine the factors that influence the development of abnormal thyrotropin (TSH) level in an euthyroid population.Methods We conducted a follow-up study in 3 communities with different iodine status.Of the 3403 euthyroid subjects at baseline screened in 1999,80.1% ( n = 2727 ) was visited and sampled in 2004 for measuring TSH,thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb).Results Iodine status in the 3 communities were stable.Decreased TSH level( <0.3 mU/L) developed in 2.5% (n =68) of sampled subjects,while raised TSH level( > 4.8 mU/L) in 2.4% (n = 64).A logistic analysis showed that risk factors for developing decreased TSH level included positive conversion of TPOAb (OR = 5.5 ),positive TPOAb both in 1999 and in 2004 ( OR = 4.0),positive TgAb in 2004 ( OR = 3.7) and TSH < 1.0 mU/L in 1999 ( OR = 2.6).Risk factors involved in developing raised TSH level included iodine status of Zhangwu community ( OR = 4.1 ),iodine status of Huanghua community ( OR = 3.9),positive TgAb in 2004 ( OR = 3.7 ),positive TPOAb both in 1999 and 2004 (OR =3.6),positive conversion of TPOAb (OR =2.7) and TSH > 1.9 mU/L in 1999 (OR = 2.6 ).Conclusions Exposure to long-term iodine excess imposes danger of developing hypothyroidism.The risk will be even higher when exposing to iodine adequacy after correction of iodine deficiency.An interval between 1.0 and 1.9 mU/L of TSH level was optimul with the least probability of developing abnormal TSH level.

OBJECTIVEEosinophilic airway inflammation is one of the basic characteristics of allergic asthma. Toll-like receptor is one of the most important innate immunity pattern recognition receptors. Glucocorticoids (GCS) are still the most effective treatment for asthma. However, few reports of studies on regulatory mechanism of GCS on the innate immunity system are available. The mechanism of effects of GCS on TLR4 is unclear. The present study aimed at understanding the effect of dexamethasone (DXM) on change of TLR4 and mechanism of regulatory effect of TLR4 on eosinophil (EOS) apoptosis.

METHODSTwenty-seven Sprague-Dawley (SD) rats (age 28 to 42 days, body weight 120 to 180 gram) were randomly divided into the control group, asthma group and DXM group with 9 in each. Asthma model rats were sensitized with the mixture of ovalbumin (OVA, 1 mg) and Al (OH)(3), 100 mg on day 1 and day 8, repeatedly exposed to aerosolized OVA after day 15, once a day for three days and continued for 30 minutes at every time. During the sensitization stage, 100 microg/ml DXM were prepared with DXM group for every other day, and the same doses DXM were prepared for every day on the stage of challenge. The histopathological changes of lung tissues were observed with light microscope (LM). EOS and other inflammatory cells in bronchoalveolar lavage fluid (BALF) were counted; the concentrations of OVA-sIgE in serum were measured by using "sandwich" ELISA; The expressions of TLR4 mRNA were determined by in situ hybridization, the apoptosis of EOS was detected by TUNEL.

RESULTS(1) LM showed many inflammatory cells infiltration around the bronchi and blood vessels, bronchus mucus increased, airway epithelium damage and desquamation, and airway mucous plugs in asthma group, whereas DXM group showed significantly milder changes. (2) Inflammationary cells count in BALF of asthma group was significantly higher as compared to control group (P < 0.01); compared with asthma group, the total cell count, EOS absolute count and EOS% were all significantly decreased in DXM group [(2.14 +/- 0.10) x 10(9)/L, (4.78 +/- 1.23) x 10(7)/L, (2.17 +/- 0.25)%]. (3) Levels of OVA-sIgE in serum of asthma group [(83.40 +/- 6.80) microg/ml] were significantly higher than those of the control group [(14.38 +/- 4.25) microg/ml] (P < 0.01), while those of DXM group [(45.02 +/- 7.47) microg/ml] were significantly lower than asthma group (P < 0.0 1). (4) There were no significant differences in TLR4 mRNA detected by in situ hybridization between control group (24.71 +/- 0.85) and asthma group (25.81 +/- 3.56) (P > 0.05); but it significantly increased in DXM group (29.86 +/- 3.92) as compared to asthma group. (5) The percentages of apoptotic EOS in asthma group [(7.39 +/- 1.93)%] were significantly lower than those in control group [(9.06 +/- 1.52)%] (P < 0.01); and significantly higher in DXM group [(13.33 +/- 1.09)%] than in asthma group (P < 0.01). There were significantly positive correlations between TLR4 mRNA and the percentage of apoptotic EOS (r = 0.612, P < 0.01).

CONCLUSIONDXM can decrease OVA-sIgE level, induce EOS apoptosis, which may correlate with the activation of TLR4 signal transduction.

Animals ; Apoptosis ; Asthma ; chemically induced ; immunology ; Bronchoalveolar Lavage Fluid ; cytology ; Dexamethasone ; pharmacology ; Eosinophils ; immunology ; Glucocorticoids ; pharmacology ; Immunoglobulin E ; blood ; Lung ; pathology ; Ovalbumin ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; immunology ; metabolism