Objective To investigate the molecular mechanism of epithelial-mesenchymal transition (EMT) induced by activated vascular endothelial growth factor receptor-1 ( VEGFR-I ) in cell line MHCC97-H.Methods MHCC97-H cells were cultured in DMEM with 1％ fetal bovine serum (control group),10 μmol/L PP2 (PP2 group),10 μmol/L PBS (PBS group),50 μmol/L VEGF-B (VEGF-B group),l0μmol/L PP2 and 50 μmol/L VEGF-B (PP2 +VEGF group),10 μmol/L PBS and 50 μmol/L VEGF-B (PBS + VEGF-B group),respectively.Protein expressions of epithelial marker E-cadherin,α-catenin and mesenchymal marker vimentin and N-cadherin were detected by Western blot.The expression sites of E-cadherin,α-catenin and mesenchymal marker vimentin and N-cadherin were detected by cell immunofluorescence.The ability of invasion and migration of cell line MHCC97-H were assessed by cell invasion and migration test.All data were analyzed by the t test.Results The expressions of E-cadherin,α-catenin,vimentin and N-cadherin were 3.23 +0.76,3.01 ±0.25,3.01 +0.22 and 2.63 +0.40 in the control group,4.18 +0.32,3.29 +0.11,4.85 +0.36 and 3.02 +0.52 in the PP2 group,2.83 +0.65,3.03 +0.27,1.37 ±0.24 and 2.98 ±0.36 in the PBS group,2.06 ±0.15,2.84 ±0.76,5.79 ± 0.38 and 5.54 ± 0.28 in the VEGF-B group,6.12 ± 0.08,5.45 ± 0.37,3.36 ± 0.42 and 3.26 ±0.13 in the PP2 + VEGF-B group and 1.36 ±0.54,1.26 ±0.45,4.05 ±0.17 and 1.05 ±0.33 in the PBS +VEGF-B group.There was a significant difference in the expressions of E-cadherin and α-catenin between the PP2 +VEGF-B group and the VEGF-B group (t =7.625,9.931,P ＜ 0.05 ).The expressions of vimentin and N-cadherin in the PP2 + VEGF-B group were significantly lower than those in the VEGF-B group (t =12.001,11.910,P ＜ 0.05).Six hours after the treatment with VEGF-B,the numbers of MHCC97-H migrated were 19 ± 1,5 ± 2and 16 ± 1 in the VEGF-B group,PP2 + VEGF-B group and PBS + VEGF-B group,respectively.The number of MHCC97-H cells migrated in the VEGF-B group was greater than that in the PP2 ± VEGF-B group ( t =13.566,P ＜ 0.05 ).The number of MHCC97-H cells passed through the Boyden chamber was 4 + 2,which was significantly less than 16 ± 1 of the VEGF-B group (t =12.350,P ＜0.05).Conclusion EMT induced by activated VEGFR-1 was mediated via c-Src kinase signal transduction in MHCC91-H cell line,and c-Src may be a potential target to interfere the invasion and migration of hepatic cancer cells.

Objective To evaluate the relationship between asthma and rhinitis in children.Methods One hundred and thirty children with rhinitis were divided into two groups.Among them,60 displayed rhinitis alone and 70 displayed rhinitis associated asthma.The following parameters were analyzed between two groups: age,sex,history of eczema,familial history of smoking,familial history of asthma,sensitization to allergens,level of total serum IgE,blood eosinophil count.Logistic regression analysis was used to analyze the effect of covariates on risk of rhinitis or asthma.Results History of bronchiolitis,familial history of asthma,maternal asthma and sensitization to allergens h_1(greer labs inc),d_2(dermatophagoides farinae) were significantly more frequent in asthmatic subjects.In these patients,the total serum IgE and eosinophil count were higher.Logistic regression analysis showed that a high eosinophil count and total serum IgE significantly increased the risk of developing asthma in patients with rhinitis.Conclusions In subjects with rhinitis,the occurrence of asthma is related to history of bronchiolitis,familial history of asthma,atopy,total serum IgE levels and blood eosinophilia.In rhinitis subjects,these parameters will be taken into account to manage underlying asthma.

OBJECTIVETo explore the molecular mechanism of brain protection against convulsive brain damage in premature brains by observing the changes of apoptotic-regulating genes of bcl-2 and c-Jun expression in the hippocampus in Wistar rats with different ages after status convulsion (SC).

METHODSSC was induced in infant Wistar rats (IRs) and adult Wistar rats (ARs) by intraperitoneal injection of lithium-pilocarpine. The rats were sacrificed at 3 hrs, 6 hrs, 12 hrs, 1 day, 3 days and 7 days after SC (n=8). Bcl-2 and c-Jun protein and mRNA levels were measured using immunocytochemistry, RT-PCR and in situ hybridization.

RESULTSc-Jun protein levels increased significantly at 3 hrs and reached the peak at 6 hrs after SC in both IRs and ARs compared to those in the normal control group (P<0.01). c-Jun protein levels started to decrease 12 hrs after SC in both IRs and ARs. The expression of c-Jun protein in IRs returned to the basal level 1 day after SC, while remained higher in ARs than in the normal control group by 7 days after SC. The expression of c-Jun protein in ARs was much higher than that in IRs from 6 hrs to 7 days after SC (P<0.05). c-Jun mRNA level was in parallel with the protein level as mentioned in IRs and ARs after SC. There were no changes observed in both bcl-2 protein and bcl-2 mRNA levels after SC in IRs and ARs.

CONCLUSIONSSC may induce an up-regulation of proapoptotic gene c-Jun in the hippocampus after SC, with a less strong extent and shorter duration in IRs compared to that in ARs. This might be one mechanism of brain protection against convulsive brain damage in IRs. The expression of bcl-2 remains unchanged after SC and is not affected by age in both IRs and ARs.

Animals ; Apoptosis ; Gene Expression Regulation ; Hippocampus ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; genetics ; Proto-Oncogene Proteins c-jun ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Seizures ; metabolism

The purpose of this study was to determine the changes of pathogens in hematological ward and susceptibility of patients received chemotherapy to antibiotics. The pathogens were taken from blood, urine and sputum of patients who accepted chemotherapy from years 2001 to 2005, then were isolated and identified. The susceptibility test was performed by disk diffusion method. The results showed that the total of 418 strains were detected. Gram-negative bacteria were the most common of nosocomial infection. Pseudomonas aeruginosa, Enterobacter cloacae, E. coli account for the most of Gram negative- bacteria infection and most resistant to broad-spectrum penicillin, Acinetobacter baumannii showed a trend of increase. The ratios of gram positive bacteria and fungi were increased slowly, mainly as Enterococcus and Candida. Enterococcus is the most common cause of Gram-positive bacterial infection. Vancomycin resistance did not occur. It is concluded that Gram-negative bacteria are main cause of nosocomial infection in patients with hematological malignancies. Gram positive bacteria and fungi had been more frequent. Strains resistant to antimicrobial agents increase.
Cross Infection ; epidemiology ; microbiology ; Drug Resistance, Bacterial ; Gram-Negative Bacteria ; drug effects ; isolation & purification ; Gram-Negative Bacterial Infections ; epidemiology ; microbiology ; Hematologic Diseases ; microbiology ; Hematologic Neoplasms ; microbiology ; Humans ; Microbial Sensitivity Tests

OBJECTIVETo investigate the risk factors causing tracheal stenosis after tracheotomy for mechanical ventilation.

METHODSA retrospective study was carried out to review the clinical data of 560 patients who had been tracheotomy for mechanical ventilation in the First Affiliated Hospital of Sun Yat-sen University from 1990 to 2006. The clinical relevant factors causing tracheal stenosis included age, sex, preoperative intubation, preoperative intubation time, postoperative mechanical ventilation duration, airway infection, multiple changes of intubation tube, cricothyroidotomy, previous tracheotomy, gastroesophageal reflux, diabetes, etc. Multivariate stepwise logistic regression model was used for the analysis.

RESULTSFifty-four cases (9.6%) presented tracheal stenosis in 560 patients after tracheotomy. With multivariate analysis, it was confirmed that the following variable correlated to tracheal stenosis. i.e, preoperative intubation time (chi2 = 4.323, P = 0.038), postoperative mechanical ventilation duration (chi2 = 14.062, P = 0.000), airway infection (chi2 = 8.604, P = 0.004), diabetes (chi2 = 5.237, P = 0.014). The effect degree of these risk factors was as below, postoperative mechanical ventilation duration (OR = 10.818), airway infection (OR = 6.349), diabetes (OR = 3.019), intubation time preoperative (OR = 2.156).

CONCLUSIONSAmong patients who received tracheotomy for mechanical ventilation, the clinical relevant factors causing tracheal stenosis were various. Statistical analysis showed that preoperative intubation time, postoperative mechanical ventilation duration, diabetes, airway infection were main risky factors which may cause tracheal stenosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Respiration, Artificial ; adverse effects ; Retrospective Studies ; Risk Factors ; Tracheal Stenosis ; etiology ; Tracheotomy ; adverse effects ; Young Adult

OBJECTIVETo investigate the clinical relevant factors causing laryngeal stenosis after partial laryngectomy.

METHODSA retrospective study was carried out to review the history clinical data from 138 patients of partial laryngectomy in the First Affiliated Hospital of Sun Yat-Sen University between January 1994 to October 2004. The clinical relevant factors causing laryngeal stenosis were included as follows: age, sex, TNM stage, tumor site, extension of thyroid cartilage defect, extension of larynx parenchyma defect, reconstruction method, laryngeal dilator, duration of using antibiotics, postoperative radiotherapy, lung infection, gastroesophageal reflux, diabetes. Multivariate stepwise logistic regression model was used for the analysis.

RESULTSOf 138 cases after partial laryngectomy, stenosis developed in 25 cases. The occurrence rate was 18.1%. In multivariate analysis, it was confirmed that the following factors correlated to laryngeal stenosis, i. e, extension of thyroid cartilage defect (chi2 = 4.323, P = 0.038), postoperative radiotherapy (chi2 = 6.002, P = 0.014), lung infection (chi2 = 4.220, P = 0.040), and gastroesophageal reflux (chi2 = 5.614, P = 0.018).

CONCLUSIONSThe clinical relevant factors causing laryngeal stenosis after partial laryngectomy were multiple. Statistical analysis showed that extension of thyroid cartilage defect, postoperative radiotherapy, lung infection and gastroesophageal reflux were the risk factors which may cause laryngeal stenosis.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Laryngeal Neoplasms ; pathology ; surgery ; Laryngectomy ; adverse effects ; Laryngostenosis ; etiology ; pathology ; Logistic Models ; Male ; Middle Aged ; Neoplasm Staging ; Postoperative Complications ; Retrospective Studies ; Risk Factors

The beta-glucosidase encoding gene bglA was cloned from Bacillus polymyxa 1.794. The bglA gene was inserted in expression vector pET28a(+) and transformed into Escherichia coli BL21 (DE3), finally the recombinant strain BL1979 was obtained. Induced by IPTG, the expression P-glucosidase activity reached to 24.7 IU/mL. The optimum temperature and optimum pH of the recombinant expression P-glucosidase in BL1979 were 37 degrees C and 7.0 respectively,the purity can reach to 92.7%. Analysis of the fusion protein by nondenaturing gradient gel electrophoresis, we found the fusion protein exists in dimmer, tetramer,hexamer and octamer, they all have hydrolase activity.
Bacillus ; enzymology ; Escherichia coli ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis ; isolation & purification ; beta-Glucosidase ; genetics ; isolation & purification ; metabolism

Abdominal Wall ; Adenocarcinoma ; diagnosis ; Colonic Neoplasms ; diagnosis ; Female ; Histiocytoma, Malignant Fibrous ; diagnosis ; Humans ; Ileal Neoplasms ; diagnosis ; Ileocecal Valve ; Leiomyoma ; diagnosis ; Middle Aged ; Neoplasms, Multiple Primary ; diagnosis ; Soft Tissue Neoplasms ; diagnosis ; Uterine Neoplasms ; diagnosis

OBJECTIVE: To investigate the role of adult cardiomyocytes in the differentiation of mesenchymal stem cells (MSCs) into cardiomyocytes. METHODS: Rat MSCs were isolated by a Percoll's gradient solution and cultured in low-glucose Dulbecco' s modified Eagle' s medium (DMEM). After 2 passages, cell-surface antigen CD34, CD71 and CD90 for rat MSCs were determined by flow cytometry, and these MSCs were transfected with pEGFP-N3 by Lipofectamine2000. Then those MSCs labeled with GFP, were cultured in contacted, nocontacted and conditioned with adult rat myocardiocytes. Immunofluorescence staining against alpha-actin, desmin, and troponin-T were performed after 1 week. RESULTS: Immunofluorescence staining was positive against alpha-actin, desmin, and troponin-T on MSCs in contacted culture group. In contrast, no alpha-actin, desmin, and troponin-T expression on MSCs were observed in the noncontacted culture group and the conditioned culture group. CONCLUSION Direct cell-to-cell contact between MSCs and adult cardiomyocytes may induce differentiation of MSCs into cardiomyocytes.
Animals ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; Cell Communication ; Cell Differentiation ; physiology ; Cell Separation ; Cells, Cultured ; Coculture Techniques ; Female ; Male ; Mesenchymal Stem Cells ; cytology ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Sprague-Dawley ; Thy-1 Antigens ; analysis

BACKGROUNDThe new Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation was developed to address the systematic underestimation of glomerular filtration rate (GFR) by the Modification of Diet in Renal Disease (MDRD) Study equation in patients with relatively well-preserved kidney function. Performance of the new equation in the Chinese population is unknown. The goal of the present study was to compare performance of these two equations in Chinese patients with chronic kidney disease (CKD).

METHODSWe enrolled 450 Chinese patients (239 women and 211 men) with CKD in the present study. The renal dynamic imaging method was used to measure the referenced standard GFR (rGFR) for comparison with estimations using the two equations. Their overall performance was assessed with the Bland-Altman method and receiver-operating characteristics (ROC) analysis. Performance of the two equations in lower and higher estimated GFR (eGFR) subgroups was further investigated.

RESULTSBoth eGFRs correlated well with rGFR (r = 0.88, 0.81, P < 0.05). In overall performance, the CKD-EPI equation showed less bias, higher precision and improved accuracy, and was better for detecting CKD. In the higher-eGFR subgroup, the CKD-EPI equation corrected the underestimation of GFR by the abbreviated MDRD equation.

CONCLUSIONSThe CKD-EPI equation outperformed the abbreviated MDRD equation not only in overall performance but also in the subgroups studied. For the present, the CKD-EPI equation appears to be the first-choice prediction equation for estimating GFR.

Adult ; Aged ; Asian Continental Ancestry Group ; Female ; Glomerular Filtration Rate ; physiology ; Humans ; Kidney Failure, Chronic ; physiopathology ; Male ; Middle Aged ; Models, Theoretical