OBJECTIVE To understand the distribution of common pathogens and their drug resistance trend and to provide the basis of the correct selection of antibiotics for clinic.METHODS Retrospective analysis was taken for blood culture specimens during 2 years in our hospital.And statistical analysis was done.All of 4028 cases of blood culture specimens were detected on automatic BacT/Alert3D rapid blood culture system,strains isolated were taken to VITEK-2 automatic microbiological analysis/sensitivity system for identification and drug susceptibility testing.Drug susceptibility results were analyzed using WHONET 5.3 software.RESULTS Totally 435 pathogen were isolated from 4028 cases of blood culture sample and the positive rate was 10.8%.195 Gram-negative bacteria strains accounted for 44.8%,mainly Escherichia coli,Klebsiella pneumoniae,Burkholderia cepacia,Acinetobacter baumannii,and Pseudomonas aeruginosa.Gram-positive bacteria accounted for 49.9%;mainly Staphylococcus epidermidis,S.haemolyticus and S.aureus dominated.Fungi were 23 strains(5.3%),mainly Candida albicans.Among them,extended-spectrum ?-lactamases(ESBLs) produced by E.coli and K.pneumoniae strains were 43.9% and 48.8%,respectively.Meticillin-resistant S.aureus and coagulase-negative staphylococci were 35.3% and 72.9%,respectively.But vancomycin-resistant S.aureus was not found.CONCLUSIONS The bacteria identification detected in blood culture class is quite complicated,and the drug resistance is high.Laboratories should increase the detection rate of bacterial culture and provide drug monitoring results for the clinics on time based on CLSI norms.Clinicians should use the antibiotics reasonably based on the drug susceptibility results in order to reduce nosocomial infections and the emergency of multiple drug-resistant strains.

Objective To evaluate the effects of hyperbaric oxygen on the expressions of hypoxia-inducible factor 1α (HIF-1α) and insulin-like growth factor-1 receptor (IGF-1R) in the formation of hyperplastic scar in rabbit ears.Methods The ears of 20 New Zealand rabbits were used to construct an animal model for hyperplastic scar by operation.After the establishment of scar models,the rabbits were randomly divided into 4 experimental groups and one control group with 4 mice (48 wound surfaces) in each group.The mice in the 4 experimental groups were treated with hyperbaric oxygen for 7,14,21 and 28 days,respectively,and those in the control group remained in normoxic environment after operation.Scar tissues were resected from all the rabbit ears on day 29 after operation.Hematoxylin and eosin (HE) staining was conducted for the observation of morphological changes and calculation of scar elevation index,and immunohistochemistry to measure the expressions of HIF-1α and IGF-1R.Statistical analysis was carried out by one-way analysis of variance followed by least significant difference t-test.Results HE staining showed that both the number of fibroblasts and amount of collagen fibers were significantly reduced in the experimental groups compared with the control group.Scar elevation index was 4.28 ± 0.22 in the control group,3.64 ± 0.29,3.46 ± 0.21,3.29 ± 0.21,3.16 ± 0.15 in the 7-,14-,21-and 28-day experimental groups respectively,with significant differences among these groups (F =77.70,P ＜ 0.05).The expressions of HIF-1α and IGF-1R were significantly lower in these experimental groups than in the control group (all P ＜ 0.01),lower in the 14-day group than in the 7-day group (P ＜ 0.05),and lower in the 21-day group than in the 14-day group (P ＜ 0.05),with no significant differences between the 28-day group and 21-day group (both P ＞ 0.05).Conclusion Hyperbaric oxygen can effectively down-regulate the expressions of HIF-1α and IGF-1R in scar tissue,and significantly inhibit the formation of hyperplastic scar in rabbit ears.

Objective: To investigate the osteointegration and osteoinduction of nano hydroxyapatite/bioglass ( nHA/BG ) gradient nanofilm on the surface of titanium ( Ti) prepared by hypotherm sintering and plastic deformation. Methods:Hypotherm sintering was used to produce nHA/BG gradient coating followed by soaking in the simulated body fluid. Ti implants with gradient coatings were planted in femoral condyles at one side of 12 New Zealand rabbits and the untreated Ti implants were planted at the other side as the controls. 1, 3 and 6 months after implantation, the animals were sacrificed after X-ray examination and the tissues around the implants from the 3 month group were used for the preparation of hard tissue section and ground section. New bone formation was observed by tetracycline fluorescence staining. Von Gieson staining was used to observe the osteointegration at the interface between bone and im-plant. Results:The gradient coatings were porous and composed of irregular rod-like nano-HA crystals. Animal study showed well es-tablished osteointegration between the gradient coating and more novel bone was found around the implants with gradient coatings. Conclusion:Osteointegration and ostioinduction of Ti implant can be enhanced by nanostructured surface with gradient coatings of nHA/BG.

Objective To compare the effects of remifentanil infused at different rates on median effective target plasma concentration (EC50) of propofol inhibiting responses to laryngeal mask airway (LMA) insertion and determine the optimum infusion rate of remifentanil when used for fiberoptic bronchoscopy in pediatric patients.Methods Eighty-four ASA Ⅰ or Ⅱ pediatric patients,aged 7 months-3 years,scheduled for elective fiberoptic bronchoscopy,were randomly assigned into 3 groups (n =28 each):normal saline group (group C),remifentanil infused at 3 ng· kg-1 ·min-1 group (group R1) and remifentanil infused at 5 ng· kg-1 · min-1 group (group R2).Responses to LMA insertion were defined as body movement and/or bucking during insertion.The initial target plasma concentrations of propofol were 5.2,4.8 and 4.4 μg/ml in groups C,R1 and R2,respectively.The target plasma concentration of propofol was determined by up-and-down sequential allocation.Each time the target plasma concentration increased/decreased by 0.2μg/ml.EC50 and 95 ％ confidence interval of propofol blunting responses to LMA insertion were determined by probit method.Results EC50 (95 ％ confidence interval) of propofol was 5.03 (4.92-5.12) μg/ml,4.71 (4.58-4.84) μg/rnl and 4.46 (4.20-4.94) μg/ml in groups C,R1 and R2,respectively.There was no significant difference in EC50 of propofol between groups R1 and C (P ＞ 0.05).EC50 of propofol was significantly lower in group R2 than in groups C and R1 (P ＜ 0.05).Conclusion The infusion rate of remifentanil should not be lower than 5 ng· kg-1· min-1 when combined with propofol in pediatric patients undergoing fiberoptic bronchoscopy.

Humans ; Laryngeal Neoplasms ; pathology ; Male ; Middle Aged ; Neurilemmoma ; pathology

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

Heart Neoplasms ; pathology ; Heart Ventricles ; pathology ; Humans ; Infant, Newborn ; Male ; Rhabdomyoma ; pathology

Objective To study the impact of tumor necrosis factor-α (TNF-α) and its antagonist on the expressions of intestinal mucosa claudin-1,Zonula Occludens-1 (ZO-1) and myosin light chain kinase (MLCK) in rat models of acute liver failure.Methods Fifty four healthy male SpragueDawley (SD) rats were randomly divided into normal control group,model group and intervention group according to a random number table.Rats in normal control (n=6) group were intraperitoneally injected with 0.9％ saline (12 mL/kg).Rats in model group (n=24) and intervention group (n=24) were intraperitoneally injected with a full dose of D-galactosamine (D-GalN) at a dose of 1 200 mg/kg to establish model of acute liver failure,while rats in intervention group were intraperitoneally injected with TNF-α antagonists (rhTNFR∶Fc) at a dose of 12.5 mg/kg before 24 hours given D-GalN.At each time point of hour 8,24,48 and 72,six rats in both model group and the intervention group were sacrificed,respectively,while the normal control group were all anesthetized and sacrificed at 72 h.Models were repeated five times.Serum liver function was detected by biochemical method,and serum TNF-α level was detected by enzyme-linked immunosorbent assay (ELISA).Hematoxylin-eosin (HE) stained sections of liver and terminal ileum were examined under an optical microscope for pathological changes;and protein expression of the terminal ileum Claudin-1,ZO-1 protein and MLCK were determined by immunohistochemistry and Western blot.Means among groups were compared with t test.Results Acute liver failure was successfully induced in the D-GalN injected rats.In the model group,alanine aminotransferase (ALT) began to decline,total bilirubin continued to rise,and enzyme-jaundice separation developed at hour 72.But total bilirubin in intervention group at hour 72 was decreased.Light microscope showed that at hour 72,villus lodged at terminal ileum in the model group with part of villus tip failing off in the model group.Villus mucosa and submucosa interstitial were edema and infiltrated with numerous neutrophils.The terminal ileum kept integrate in the intervention group,and villus mucosa and submucosa were mild edema and only infiltrated with a small amount of neutrophil.Expressions of tumor necrosis necrosis factor (TNF)-α in rats of model group and intervention group were gradually increased and peaked at hour 24 ([239.83 ± 15.81] and [182.71± 17.08] ng/L,respectively),which were significantly higher than that of the control group ([24.19±3.57] ng/L,t=22.68and 15.73,respectively;both P＜0.01).Expression of serumTNF-α in the intervention group was significantly lower than that of model group (t=4.58,P＜0.01).Expressions of Claudin-1 and ZO-1 in model group decreased gradually at an early stage and reached the lowest level at hour 24 (0.355 ± 0.068 and 0.387 ± 0.091,respectively),which were both significantly lower than that of control group (1.640±0.188 and 1.015±0.150,respectively;t=12.87 and 7.14,respectively;both P＜0.01).In the intervention group,expressions of Claudin-1 and ZO-1 also decreased to the lowest level at hour 24 (1.051 ± 0.370 and 0.642 ± 0.082,respectivley),which were both significantly lower than that of control group (t =2.84 and 4.36,respectively;both P＜0.05),but significantly higher than model group with stastically difference (t =3.70 and 4.15,respectively;both P＜0.01).MLCK protein levels in the model and intervention group were gradually increased,which peaked at hour 24 (1.298±0.194 and 1.033 ± 0.073,respectively),significantly higher than the control group (0.460±0.069,t=8.16 and 11.44,both P＜0.01);and MLCK in the intervention group was lower than that in the model group with statistically difference (t=2.56,P＜0.05).Conclusions Expression of serum TNF-α in rat model of acute liver failure increases,which leads to decreased expression of Claudin-1 and ZO-1,and increased expression of MLCK,makes cell shrunk and cell gap increased.TNF-α antagonist could significantly reduce the inflammation and liver cell apoptosis,improve liver function by inhibiting MLCK expression and preventing decrease of Claudin-1 and ZO-1 proteins.

Objective To evaluate the clinical efficacy of tissue-type plasminogen activator (tPA) treatment of children with plastic bronchitis.Methods The study retrospectively reviewed the clinical data of the children with plastic bronchitis who were admitted to Tianjin Children's Hospital from September 2013 to January 2015 and were treated with tissue-type plasminogen activator.This study analyzed the effect and safety of tPA treatment,including clinical and radiological changes and follow-ups.Results A lot of plastic secretions were safely removed from the bronchial tubes in all children and clinical manifestations including breathing,body temperature,transcutaneous oxygen saturation and image changes were significantly improved.Conclusions Bronchoscopy is an effective way to treat plastic bronchitis,but with the use of tPA a better clinical efficacy could be achieved.The method is safe and effective and should be applied early in the patients in order to prevent the occurrence of severe airway obstruction complications.

Purpose To investigate the serum prostate specific antigen( PSA)feature of high grade prostatic intraepithe1ia1 neop1asia ( HGPIN)patients,and the association of the number of cores positive for HGPIN on initia1 biopsy and the risk of cancer deve1opment in second biopsy. Methods 492 cases of patients with suspicious prostate cancer were schedu1ed for transrecta1 u1trasound prostatic biopsy with an 8-core temp1ate. In the first biopsy,186 cases of patients with PCa,34 cases of patients with iso1ated HGPIN( on1y one core invo1ved with HGPIN)and 13 cases of patients with extensive HGPIN( two or more cores invo1ved with HGPIN),64 cases of pa-tients with LGPIN,195 cases of patients with BPH. The va1ues of PSA were ana1yzed and compared within these groups. In patients with extensive HGPIN or iso1ated HGPIN we proposed a repeat 8-core biopsy after 6 months independent of serum PSA 1eve1. The same measure was app1ied for patients diagnosed as LGPIN or BPH in the first biopsy with accompanying increase or persistent e1evation of serum PSA 1eve1. The incidence of PCa was ana1yzed and compared within these groups. Results The serum PSA 1eve1s were no sig-nificant1y different between LGPIN and BPH(P>0. 05),between iso1ated HGPIN and LGPIN(P>0. 05),and between iso1ated HG-PIN and BPH(P>0. 05). The serum PSA 1eve1s were significant1y different between extensive HGPIN and LGPIN(P<0. 05),be-tween extensive HGPIN and BPH(P<0. 05),and between extensive HGPIN and iso1ated HGPIN(P<0. 05). In the second biopsy, the incidence of PCa in patients with extensive HGPIN was 38. 48%,that in patients with iso1ated HGPIN was 9. 68%,that in patients with LGPIN was 12. 50%,and that in patients with BPH was 12. 20%. Conclusions The features of PSA in patients with iso1ated HGPIN are simi1ar to BPH,PSA 1eve1 in patients with extensive HGPIN were between PCa and BPH,and patients with extensive HG-PIN have a higher incidence of PCa in second biopsy than iso1ated HGPIN and BPH.