Objective To study the expression of basic fibroblast growth factor(bFGF)in the kidney tissues of chronic interstitial nephritis(CIN).Methods Expression of bFGF was assayed in 30 patients with CIN and 5 normal persons by SP method of immunohistochemical technique.The average gray value and average optical density of the bFGF positive parts and its ratio with the whole visual field had been measured with video analysis system.Results The expression of bFGF in the kidney tissues of CIN was significantly stronger than that in normal tissues.The average gray value,average optical density and the positive areas were significantly high compared with that of normal cases.Conclusion The bFGF can promote the proliferation of fibroblasts and monocytes in renal interstitial patients and lead to renal interstitial fibrosis in the end.

ObjectiveTo observe the effect of rehabilitation on fractures around knee treated with external fixation.Methods48 cases with the fractures around knee who accepted early rehabilitation after external fixation had been observed for 0.5～4 years.ResultsAll the cases came up to clinical healing standards within 7 months. According to Kolmert's standard, the rate of fine and good was 89.6%.ConclusionThere was satisfactory outcome treating fractures around knee with external fixation. Early rehabilitation is important to avoid the knee joint rigor.

Objective To study the clinical value of detection of peripheral blood natural killer(NK) cells subsets in patients with malignant hematological diseases.Methods A total of 100 patients with malignant blood disease were enrolled from March 2014 to April 2016 as the observation group,including 50 cases in acute stage and 50 cases in mild stage.At the same time,100 health subjects were enrolled as the control group.The number of NK cells in peripheral blood and the changes of subgroups were compared.Results The number of NK cells in the different groups were compared,the absolute numbers and relative numbers of NK cells in the acute phase of leukemia and lymphoma were much lower than those in the control group(P<0.05).While the absolute number and relative number of NK cells in mild stage of leukemia and lymphoma were higher than that in acute phase(P<0.05).After treatment,the absolute number and relative number of NK cells in patients with leukemia and lymphoma were higher than before treatment(P<0.05).Before treatment,levels of CD56bright and CD56dim in patients with leukemia and lymphoma were statistically different with those of control group(P<0.05).After treatment,CD56bright and CD56dim levels were significantly improved in both groups(P<0.05).Conclusion The number of peripheral blood NK cells in patients with hematological malignancies could be reduced,and the cells function might be compromised.Detection of NK cell subsets could be with guiding significance in the treatment of malignant hematologic disease.

Objective To determine the involvement of DNA demethylation in tumor necrosis factor-α(TNF-α)-induced matrix metalloproteinase 9 ( MMP9) expression in human epidermal keratinocytes. Methods Real-time RT-PCR, Western blot, and enzyme-linked immuno sorbent assay (ELISA) were performed to determine the mRNA and protein levels of MMP9 after human keratinocyte cell line (HaCaT) cells were treated with 10 ng/ ml TNF-α or 2. 5 μmol/ L DAC/ 300 nmol/ L TSA. Bisulfite sequencing PCR ( BSP) and Methylation-sensitive high-resolution melt analysis ( Ms-HRM) were used to detect significantly differentially demethylated CpG sites in the human MMP9 promoter region in cells exposed to TNF-α. Different sites methylation constructs of promoter-luciferase reporter gene were made to detect the influences of site-specific DNA demethylation on transcription activity of MMP9 promoter. Results Compared with PBS-treated control, TNF-α significantly increased the expression of MMP9 in HaCaT cells for indicated culture duration ( P < 0. 05 ). Real time PCR, Western blot, and ELISA analysis demonstrated that the mRNA and protein levels of MMP9 were increased initially, followed by a decline with prolonged incubation time. After TNF-α treatment, varied degrees of DNA demethylation occurred at 10 CpG sites in the promoter of MMP9, and the changes at the -36 bp site were statistically significant (P<0. 05). The demethylation at the -36 bp site greatly increased the transcription activity of MMP9. Conclusion TNF-α promotes MMP9 expression in HaCaT cells through inducing -36 bp site DNA demethylation on the promoter of MMP9.

Objective To investigate the influence of perioperative continuing aspirin therapy on tooth extraction surgery in elderly patients with coronary heart disease.Methods A total of 926 patients with coronary heart disease who required tooth extraction were enrolled in this study.Among them,241 patients discontinued aspirin therapy before tooth extraction,685 patients continued aspirin therapy before tooth extraction.The entering rate of operation was evaluated.100 patients continuing aspirin and 100 patients discontinuing aspirin were randomly selected.Systemic pressure,diastolic pressure and heart rate before and after tooth extraction were detected.100 matched healthy elderly patients were selected as normal control group.Multivariate logistic regression analysis was used to estimate the risk for tooth extraction.Results The entering rate of operation was 67.9％ (131 cases) in 193 patients with angina pectoris discontinuing aspirin therapy and 82.1 ％ (312 cases) in 380 patients with angina pectoris continuing aspirin therapy (x2 =14.77,P＜0.01).The entering rate of operation was 41.7％ (20 cases) in 48 patients undergoing coronary revascularization and discontinuing aspirin therapy,and 80.7％ (312 cases) in 305 patients undergoing coronary revascularization and continuing aspirin therapy (x2=33.95,P＜0.01).The changes in systolic blood pressure before versus after dental extraction had a significant difference between the discontinuing aspirin group and control group [(15.9±5.5) mmHg vs.(12.2±4.7)mmHg,P＜0.05,1 mmHg=0.133 kPa],while had no significant difference between the continuing aspirin group and control group [(13.6±4.5) mmHg vs.(12.2±4.7) mmHg,P＞0.05].There were no significant differences in changes in diastolic blood pressure and heart rate in the intraoperative period between the patients and controls (P＞0.05).The bleeding rate in patients continuing aspirin therapy was not significantly different as compared with control group (P＞ 0.05).Multivariate logistic regression analysis revealed that perioperative discontinuing aspirin therapy remained the second significant risk factor for tooth extraction.Conclusions Preoperative administration of aspirin should be continued in dental extraction surgery in elderly patients with coronary heart disease.

Objective To explore the effects of lead acetate on free radicals and lipid peroxidation in the cerebral cor?tex, cerebellum, and hippocampus in rat brains. Methods SD rats (n=48), who were just weaned, were randomly divided in?to 4 groups base on their weight. Then the rats were fed with lead acetate in drinking water at the final concentrations of 0 mg/L (deionized water), 200 mg/L, 400 mg/L, 800 mg/L respectively. Blood lead level as well as the hydroxyl free radical inhibiting activity, the levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA) in cerebral cortex, cerebellum, and hippocam?pus were measured 60 days after lead contamination in water. Results Upon lead exposure, blood lead levels increased sig?nificantly as compared with the control. The hydroxyl free radical inhibiting activity in cerebral cortex, cerebellum, and hip?pocampus decreased significantly in a dose dependent manner of lead(P < 0.05). And they all correlated negatively with blood lead level (r=-0.505,-0.414,-0.448, P<0.05). By contrast, blood lead level was positively correlated with H2O2 and MDA in these brain tissues (r=0.301, 0.411, 0.378, and 0.404, 0.324, 0.510,P < 0.05). Conclusion Lead exposure can lead to lipid peroxidation of rat brain tissues through inducing free radicals.

Objective Evaluate the outcome of aortic root reconstruction on the analysis of the risk factors influencing surgical results. Methods Between August 1996 and November 2009, 92 patients(56 men, 36 women) aged from 14 to 77years [mean (44.8 ±1.4) years] with aortic root aneurysm underwent aortic root reconstruction. 72 patients had over moderate aortic valve insufficiency. 47 patients suffered from Marfan syndrome. The aortic pathology was aortic dissection in 45. Bentall technique was used in 59 patients, the button technique in 13, the David I with the Valsalva graft in 6 patients and the aortic valve resuspension in 14 patients. Results The hospital mortality rate was 8.7%. The major complications 31. 7%. 18patients died during the period of follow-up. Late complications among 55 survivors were 12. Univariate predictors of the morbidity were the presence of male, non-Marfan, concomitant procedure, deep hypothermia cardiac arrest, aortic cross clamp time and blood infusion. Risk facts for mortality were emergent or urgent operation, aortic dissection, concomitant procedure, aortic cross clamp time and blood infusion. Multivariate analysis revealed risk factors of concomitant procedure and blood infusion were responsible for both morbidity and mortality. The overall long-term survival rate is (97.1 ±2.0)% at 1-year, (88.1 ±4.7)% at 5-year, (54.0 ±9.2)% at 10-year. The mean for survival time is (9.9 ±0.59) years, 95% confidence interval 8.70 -11.01. Conclusion The aortic root restitution procedures are safe and effective in general. The short and long-term outcome is satisfactory. The button technique is the first choice for reimplantation coronary patch. Valve-sparring aortic root reconstructions show promise in safety and applicability.

Objective To investigate whether octreotide,as somatostatin analogue,can enhance the sensitivity of the human lung adenocarcinoma cell line A549 to chemotherapeutic drugs.Methods Different concentration of octretide,cisplatin and 5-Fluorouracil (5-Fu) was respectively acted on the lung adenocarcinoma cell line A549.The absorbance value was tested by colorimetry through MTT method to evaluate the effect of octreotide,cisplatin,5-Fu or the three drugs combined respectively after 48 hours.Each drug concentration had six holes and it repeated three times.The effects of combination therapy was analysed with isobologram.Results It was proved that octreotide could inhibit the proliferation of A549 cells in a dose-dependent manner at the concentration range of 1.3 mg/L ～ 166.7 mg/L.The inhibition rate was dose-dependent which was higher when octreotide combined with cisplatin and 5-Fu than it alone.It has statistically significant difference (P ＜ 0.05 ).The effect plots of IC50 were located in the synergy areas of isobologram.Conclusion It can be concluded that octreotide could inhibit the proliferation of A549 cells in vitro.This inhibition enhances when octreotide is combined with cisplatin and 5-Fu.Octreotide can enhance the susceptibility of A549 cells to cisplatin and 5-Fu.

Objective To assess the subgenotypes of pathogenic Cryptococcus gattii isolates from China and to elucidate the epidemiological links between these domestic isolates and those from other parts of the world. Methods DNA was extracted from 9 clinical isolates of Ctyptococcus gattii from China. The partially variable regions of the three unlinked loci, namely IGS1, PLB1 and GEF1, were amplified and sequenced, and the bioinformation at these loci was obtained from GenBank for multi-locus sequences alignment and phylogenetic analysis. Results Of these 9 clinical isolates, 8 were genotype VG Ⅰ and mating type α with the same sequences at the tested regions as the reference strain WM276, which was a representative isolate of an independent subgenotype; 1 was of genotype VG Ⅱ and mating type α, which was the first report in China, with the tested sequences consistent with those of the referrence strain R272. Sequencing and phylogenetic analysis of GEF1 gene, which was located at mating type locus, successfully identified the genotypes and mating types of all the Cryptococcus gattii isolates involved here. Conclusions Multi-locus sequence analysis shows that causative Cryptococcus gattii isolates of genotype VG Ⅰ in China carry similar sequences at the tested loci in IGS1, PLB1 and GEF1 genes, to a widely distributed subgenotype in the world, and the sequences of the first VG Ⅱ genotype isolate from China resemble the less virulent subgenotype VG Ⅱ b found in Vancouver islands.

BACKGROUND:Skeletal muscle satel ite cells are totipotential stem cells with multi-directional differentiation potential, locate in skeletal muscle interstitium, have a certain tolerance to ischemia and hypoxia, and are important cells in stem cellengineering.
OBJECTIVE:To establish a thrifty, convenient culture procedure and create a simple, efficient method to transfect skeletal muscle satel ite cells, and investigate genetic expression after the transfection for cellular cardiomyoplasty.
METHODS:Skeletal muscle satel ite cells were isolated from rabbit thigh and cultured. Their growth curves were determined by CKK-8 method. Grouped by different proportions of the plasmid and liposome, skeletal muscle satel ite cells were transfered by the enhanced green fluorescent protein plasmid based on liposome. After transfection, the efficiency and character of target genetic expression was determined.
RESULTS AND CONCLUSION:Satel ite cells were isolated, cultured and transfected successful y. In suitable ratio of plasmid and liposomes, the transfection efficiency reached up to above 35%. The target protein was expressed within 12 hours after transfection, reached maximum in 48-72 hours and decreased gradual y after one week. The expression stil could be observed two weeks latter. The enhanced green fluorescent protein plasmid conducted by cationic liposome could be transfered into skeletal muscle satel ite cells efficiently. The transfection efficiency was correlated closely to the ratio of plasmid and lipofectamine. The change of target gene expression depended on time.