OBJECTIVE To study Pseudomonas aeruginosa infection in respiratory tract of severe neurosurgery patients after tracheotomy,and discuss the prevention and management of nosocomial infection.METHODS The data of 19 severe patients admitted in neurosurgery from Dec 2005 to Oct 2007 who underwent tracheotomy were analyzed retrospectively.RESULTS Respiratory tract infection occurred in all 19 cases,the incidence rate was 100%.Eleven species and 73 strains of pathogenic bacteria were found.Thirty(41.10%)strains of P.aeruginosa were found in 12(63.16%)cases,the drug resistance rate was higher.CONCLUSIONS P.aeruginosa is the most important pathogenic bacteriaum of respiratory tract nosocomial infection in severe neurosurgery patients after tracheotomy.Strengthening the sterilization,isolation and airway management and selecting antibiotics based on drug sensitive test are the important methods to prevent and control infection.

Objective To investigate the correlation and characteristic of the functional polymorphism of the dopamine D4 receptor gene on the behavioral problems in school-age twins.Method The behavioral problems were assessed among a total of 147 twin pairs aged 6-12 years with the Achenbach Child Behavior Checklist (CBCL).The DRD4 gene polymorphism was applied for different types with polymerase chain reaction.The generalized estimating equations model was used to analyze the effect of DRD4 gene polymorphism and the interaction of DRD4 gene polymorphism with children age,gender,monozygotic/dizygotic on twins' behavioral problems.Results (1) School-age children's anxiety/depression,withdrawn,thought problems,attention problems,social problems,delinquent problems,aggressive behavior and total variance mainly were correlated with DRD4 gene polymorphism (P＜0.05),and physical discomfort might be related with DRD4 gene polymorphism (P=0.067).Except social problems,the detection rate of behavioral problems of the twins with 4/6 genotype (Gene type carrying 5 or more repeated sequences) of DRD4 gene (6.25％-21.25％) was higher than those with other genotypes (0-14.74％).(2) Withdrawal was related to children age (x2=4.348,P＜0.05),and twins aged 6-10 might lead to withdrawal.(3) The interaction of DRD4 gene polymorphism and children age had a significant effect on withdrawal (x2=4.348,P＜0.05) and physical discomfort(x2 =5.189,P＜0.05),and the elder age (11-12 years) children had a certainly protective effect on the withdrawal and physical discomfort with 4/6 genotype in twins.The detection rates were 3.54％ and 2.65％ in the younger-age (6-10 years) twins,while all 0 in the elder-age (11-12 years) for the withdrawal and the physical discomfort respectively.Conclusions The DRD4 gene polymorphism may be correlated with behavioral problems in school-age twins.The interaction of DRD4 polymorphism and children age had a significantly effect on withdrawal and physical discomfort.

Objective To investigate the factors of Alzheimer’s Disease (AD). Methods A 1:1 matched case-control study was conducted. The study included 129 patients of clinically diagnosed AD and 129 controls selected from Nanchang and Jiujiang areas. The data were analyzed by the conditional logistic regression. Results The results showed the unsociable personality (OR=2.486,95%CI=1.075～6.638),the history of great negative life events (OR=4.125,95%CI=1.706～12.249),using cookers made of aluminum (OR=2.454,95%CI=1.277～5.268) were identified as risk factors for AD. The good family income state (OR=0.324,95%CI=0.106～0.828) and frequently physical exercises (OR=0.258,95%CI=0.104～0.520) were protective factors for AD. Conclusion Alzheimer’s Disease seems to be related to the income state,frequency of physical exercises,personality,big negative life events,using cookers made of Aluminum.

OBJECTIVE:To investigate the drug use in the inpatients of our hospital and evaluate its rationality.METHODS:By using PASS-nested "Army Health No.1" hospital information system,the drug utilization in these patients was analyzed.RESULTS:Among 32 834 medication orders for 586 inpatients,732(1.50%)were warning ones.CONCLUSION:The drug utilization in the inpatients of our hospital was rational on the whole.PASS satisfies clinical doctors' needs,and the physicians feel satisfied with the system.Nevertheless,there are some problems remain to be tackled.

Objective To investigate the role of nucleotide-binding oligomerization domain-containing protein 1 (NOD1) in inducing insulin resistance in differentiated human adipocytes.Methods Human preadipocytes obtained via liposuction were induced to differentiate into mature adipocytes.iE-DAP,a specific ligand for NOD1,was administered to human adipocytes in culture.NF-κB transcriptional activity and proinflammatory cytokines production were determined by luciferase assay and enzyme-linked immunosorbent assay.Glucose uptake in adipocytes was measured by 2-deoxy-D-[3 H] glucose uptake (P＜0.05).The expression of phosphatidylinositol-3-kinase p85 (PI-3K p85),insulin receptor substrate-1 (IRS-1) and Akt phosphorylations were detected by Western blotting.Results NF-κB transcriptional activity and cytokines such as interleukin (IL)-6,IL-8,and monocyte chemotactic protein 1 secretion were markedly increased after stimulation with iE-DAP (P＜0.05 or P＜0.01).Insulin-induced glucose uptake was decreased with the activation of NOD1 in a dose-and time-dependent fashion.NOD1 activation weakened insulin signal transduction as being revealed by increasing IRS-1 Ser307phosphorylation,reducing protein expression of PI-3K p85,and attenuating insulin-induced phosphorylation of Akt on Ser473 and Thr308in human adipocytes.Conclusion These results indicate that NOD1 activation induces inflammatory response and insulin resistance via IRS-1/PI3-K/Akt signaling pathway in human adipocytes.

Bacillus subtilis is a generally recognized as safe (GRAS) strain that has been widely used in industries including fodder, food, and biological control. In addition, B. subtilis expression system also plays a significant role in the production of industrial enzymes. However, its application is limited by its low sporulation frequency and transformation efficiency. Immense studies have been done on interpreting the molecular mechanisms of sporulation and competence development, whereas only few of them were focused on improving sporulation frequency and transformation efficiency of B. subtilis by genetic modification. The main challenge is that sporulation and competence development, as the two major developmental events in the stationary phase of B. subtilis, are regulated by the complicated intracellular genetic regulatory systems. In addition, mutual regulatory mechanisms also exist in these two developmental events. With the development of genetic and metabolic engineering, constructing genetic regulatory networks is currently one of the most attractive research fields, together with the genetic information of cell growth, metabolism, and development, to guide the industrial application. In this review, the mechanisms of sporulation and competence development of B. subtilis, their interactions, and the genetic regulation of cell growth were interpreted. In addition, the roles of these regulatory networks in guiding basic and applied research of B. subtilis and its related species were discussed.
Bacillus subtilis ; genetics ; physiology ; Gene Regulatory Networks ; Metabolic Engineering ; Spores, Bacterial ; physiology

Objective To investigate the in vivo effect of silenced actin-associated protein Transgelin on the growth of human pancreatic carcinoma xenograft in nude mice.Methods Human pancreatic cancer cell line BxPC3 were transfected with small hairpin RNA (shRNA) eukaryotic expression vector targeting Transgelin gene.RT-PCR and Western blot were used to analyze Transgelin expression after transfection.24 animal models were randomly divided into three groups with 8 in each:Experimental group (transplanted BxPC3/Transgelin shRNA),negative control group (transplanted BxPC3/Neo) and untreated group (transplanted BxPC3).Tumor size was measured weekly.All mice were sacrificed after 28 days.Tumor volume was calculated,inhibitory effect was analyzed.Immunohistochemical staining of paraffin sections for Transgelin and proliferating cell nuclear antigen (PCNA) proteins were performed.Results Tumors varied in sizes among 3 groups (all P ＜ 0.05).On day 21 and 28 tumor was significantly smaller in experimental group than those in control groups.Tumor weighed(0.74 ±0.21) g in experimental group,lower than that in negative control group(1.42 ± 0.28) g and untreated group(1.59 ± 0.24) g (all P ＜ 0.05).The inhibitory effect was 53.5％ in experimental group.The PCNA index was significantly lower in experimental group than those in control groups (all P ＜ 0.05).Conclusions Deletion of Transgelin gene can significantly inhibit the proliferation and tumor growth of BxPC3 cells in nude mice.

BACKGROUND:The hyperpoladzation-activated cyclic nucleotide-gated cation channel (HCN) gene is not increase the risk of induced cardiac arrhythmia,and can accept the modulation function of autonomic nervous system.Therefore,it is the first candidate gene for biological pacemakers.OBJECTIVE:To construct recombinant adenovirus vector containing human HCN4 gene and evaluate its transfection efficency in rat bone marrow mesenchymal stem cells (MSCs).DESIGN,TIME AND SETTING:The in vitro cytology-gene experiment was performed at the Lin Bai-xin Experimental Center,Second Hospital of Sun Yat-sen University from February to September 2008.MATERIALS:Ten SD rats were supplied by the experimental animal center of Sun Yat-sen University.Rlasmid pcDNA3.1-HCN4 containing target gene human HCN4,human embryo kidney 293 cells and Escherichia coll DH5α were preserved by our experimental center;Shuttle plasmid pShuttle-CMV containing green fluorescent protein gene and adenoviral backbone plasmid pAdxsi were bought from SinoGenoMax Co.,Ltd.METHODS:HCN4 cDNA segment was liberated from the cloning vector of pcDNA3.1-HCN4 via Hind Ⅲ+Xba Ⅰ,and subcloned into pShuttle-CMV,which was digested by I-Ceu Ⅰ +I-Sce Ⅰ double enzyme and subcloned into adenoviral plasmid to form recombinant adenovirus plasmid.Recombinant adenovirus plasmid was transfected into 293 cell lines by liposome,and the recombinant adenovirus AdHCN4 was packaged and transfected into rat MSCs.MAIN OUTCOME MEASURES:The identification of recombinant adenovirus plasmid vector,identification of recombinant adenovirus and its titration test;the transfection efficiency of recombinant adenovirus.RESULTS:Cloned sequence about 3.6kbp was obtained by Hind Ⅲ+Xho Ⅰ digestion after HCN4 cDNA segment was cloned into pShuttle-CMV.DNA sequencing results indicated that the clone location was correct.Recombinant adenovirus plasmid was cut into seven fragments while empty vector gained only six fragments after digested by Xho Ⅰ.The recombinant adenovirus was pathogenic to 293 cells after recombinant adenovirus plasmid was packaged in it.HCN4 cDNA (657bp) was amplified by PCR with virus titer of 2.5×10~(11) PFU/mL after transfected 293 cells with supematant.The efficiency of recombinant adenovirus infecting rat MSCs was about 90% when multiplicity of infection was 800.Rat MSCs expressed green fluorescence after transfection.CONCLUSION:The adenovirus vector with human HCN4 cDNA was established successfully,which can effectively transfect rat MSCs.

Objective To explore the clinical characteristics, treatment and prognosis of pediatric patients with bronchiec-tasis caused by Mycoplasma pneumonia (MP) and Chlamydia pneumonia (CP). Methods The clinical data from 27 MP and CP pneumonia pediatric patients with bronchiectasis suggested by the high resolution CT were retrospectively analyzed. Results The morbility rate of bronchiectasis caused by MP and CP pneumonia is 0.56%. The mean age of these patients was 75.4 ± 52.7 months. Among them, 27 cases (100%) had cough, 19 cases (70.4%) had fever, 10 cases (37%) had respiratory distress and 20 cases had lung auscultation. Sixteen cases were MP-IgM positive, 5 cases (18.5%) were CP-IgM positive and 6 cases (22.2%) were positive of both. Eight cases were combined with other pathogens infections, in which 6 cases were bacterial infections. The imaging ifndings showed diffuse bronchiectasis in 13 cases (48.1%) and local bronchiectasis in14 cases (51.9%). The bronchoscopy found endothelium in-lfammation, mucosal swelling, partial erosion and follicular hyperplasia in 16 cases (66.7%), the formation of short column sputum bolt in 5 cases (20.8%), in which 1 case had plastic bronchitis. All patients were treated with macrolides antibiotics, 10 cases (37%) combined with methylprednisolone, 3 cases (11.1%) combined with immunoglobulin and 20 cases (74%) combined with other anti-biotics. The average length of hospitalization was 12±4.3 days. The bronchiectasis sign disappeared within 4 months in 23 cases (92%). Two cases (8%) still had bronchiectasis after 9 to 15 months, with the recurrent pneumonia. Conclusions MP and CP pneumonia can lead to acute bronchiectasis. Most of patients are recoverable with effective treatment.

Objective To investigate the protective effects and mechanisms of granulocyte-colony-stimulating factor (G-CSF)on a rabbit model of chronic myocardial ischemia.Methods Myocardial ischemia models were created by partial ligation of the left anterior descending coronary artery in Japanese white male rabbits.Rabbits were subcutaneously injected with G-CSF (G-CSF group)or saline (control group)for 6 days after myocardial ischemia.The percentage of CD34-positive cells in the peripheral blood was evaluated by flow cytometry,and CD34-positive cells homing and vWF expression in the ischemic myocardium were determined by immunohistochemistry.Results Rabbits in G-CSF group had a higher survival rate than those in control group (P <0.05).Immunohistochemistry of the ischemic myocardium showed that compared with control group,G-CSF group had increased homing of CD34-positive cells on day 7 post-surgery,and more vessels on day 28 post-surgery by anti-von Willebrand factor staining.In addition,we observed an increase in the percentage of CD34-positive cells in the peripheral blood in G-CSF group.Conclusion G-CSF produces an obvious protective effect against chronic myocardial ischemia in rabbits by increasing stem cell mobilization,homing to ischemic myocardium and accelerating neovascularization.