Objective To study the isolation and antimicrobial resistance of pathogens isolated from patients with brain damage in hyperbaric oxygenation department,so as to provide reference for clinical anti-infective treatment.Methods Bacterial culture and antimicrobial susceptibility testing results of pathogens isolated from blood,sputum,and urine specimens of 975 patients with brain damage in the hyperbaric oxygenation department of a hospital between January 2013 and December 2014 were analyzed retrospectively.Results A total of 1 328 strains of pathogens were detected,877(66.04%)of which were gram-negative bacteria,213(16.04%)were gram-positive bacteria,and 238(17.92%)were fungi.The top five isolated pathogens were Pseudomonas aeruginosa,Klebsiella pneumoniae,Escherichia coli,Acinetobacter baumannii,and Candida albicans.Specimens mainly isolated from sputum and urine,accounting for 58.59%and 35.24%respectively,resistance rates of Klebsiella pneumoniae,Pseudomonas aeruginosa,Acinetobacter baumannii,and Escherichia coli to imipenem were 16.67%,81.82%,82.44%,and 4.65%respectively.Vancomycin-resistant strains was not found among gram-positive bacteria,resistance rates of Enterococcus faecalis to most antimicrobial agents were lower than those of Enterococcus faecium.Conclusion Respiratory and urinary tract infection account for most of the infection in patients with brain damage in hyperbaric oxygenation department,gram-negative bacteria are the predominant pathogens causing infection.

Child ; Child, Preschool ; Constipation ; diagnosis ; physiopathology ; therapy ; Digestive System ; microbiology ; physiopathology ; Gastrointestinal Hormones ; physiology ; Gastrointestinal Motility ; physiology ; Humans

Objective To clone the telomere-associated protein T-STAR and study the relationship between T-STAR and the telomerase catalyzed subunit hTERT in mammalian cells. Method The expression vector pGBKT7-hTERT was constructed and acted as a decoy in cDNA library screened by yeast two-hybrid technology. Recombinant vectors pVP16-T-STAR and pM-hTERT were constructed and co-transfected with report gene CAT into SGC-7901 cells with liposome. pM-53+pVP16-T and pM3-VP16 were introduced as positive controls, pM-53+pVP16-CP as negative control, and pM-hTERT+pVP16, pM+pVP16-T-STAR and pM+pVP16 as crossing negative controls. The expression of CAT was assayed by ELISA. Results The eukaryotic expression vector pGBKT7-hTERT was successfully constructed. One positive clone achieved by cDNA library screening was sequenced and compared with the isogenous sequences in GenBank by Blast software via Internet. As a result, T-STAR, a recorded cDNA sequence was obtained. The recombinant vectors of pVP16-T-STAR and pM-hTERT were constructed successfully and co-transformed into gastric cancer cells SGC-7901. The OD value of reported gene CAT was 0.258, which was significantly higher than that of the negative and crossing negative controls (0.002-0.015). It revealed that T-STAR interacted with hTERT in the mammalian cells. Conclusions T-STAR interacts with hTERT in the gastric cancer cells. T-STAR may be a new member of telomere-associated protein, and it participates in the regulation of telomerase through hTERT.

OBJECTIVE:To monitor the antimicrobial resistance of Enterococcus faecalis and Enterococcus faecium in our hospital from 2006 to 2008 in order to provide reference for clinical rational use of antibiotics.METHODS:The susceptibility of isolated enterococci was determined by K-B paper diffusion method,and the antibiotic resistance rates of Enterococcus faecalis and Enterococcus Faecium were analyzed using Whonet 5 software.RESULTS:Vancomycin was the most sensitive antimicrobial against Enterococcus faecalis and Enterococcus faecium.E.faecalis had low resistance rates to penicillin G,high concentration gentamicin,ciprofloxacin and rifampicin,while E.faecium had low resistance rates to high concentration streptomycin,tertracycline,chloromycetin,and qinupristin/dalforpristin.CONCLUSION:E.faecalis and E.faecium are multi-drug resistant.Antimicrobials should be used rationally based on the susceptibility test of E.faecalis and E.faecium.

Objective To observe the characteristics of chromatin conformation in hypothalamus and pituitary gland of rats with different ages (16 17 weeks and 100 105 weeks of age). Methods Micrococcal nuclease (MCN) and deoxyribonuclease I ( DNase I) were used as the probe to differentiate chromatin conformation and agarose gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) were employed to investigate the changes of chromatin conformation in rats with different ages. Results Chromatin DNA of hypothalamus and pituitary gland in old rats existed a repeat length (nucleosome core and linker region) of (190?7)bp and (171?8)bp, and in young rats (193?9)bp and (170?5)bp respectively. PAGE showed that DNase I cleaved nucleosome DNA at 10 bp intervals and the cleavage patterns were the same in all ages of rats; comparison of DNA fragments digested by DNase I in young and old rats showed that less fragments with lower base pair were produced in old rats. Conclusions (1) No aging related changes were observed with the repeat length of chromatin nucleosome DNA in hypothalamus and pituitory gland, but there was difference depending on tissues. (2) The chromatin DNA mainly existed in B type of duplex conformation and contained similar super helical structure of solenoid in hypothalamus. (3) Further experiments showed that the chromatin DNA in hypothalamus and pituitary gland from elderly rats was more resistent to DNase I digestion.

Objective To study and observe the engraftment of donor cells from unrelated cord blood into adult patients with severe aplastic anemia (SAA) and the outcome of allo-CBSCT. Methods Six patients received cord blood provided by Guangzhou Cord Blood Bank, for three of which one unit of cord blood was given in a procedure, whereas for other 3 patients, 2 units of cord blood were infused at the same time in a transplant protocol. In all 9 units of umbilical cord blood (UCB) infused, UCB units contained 1.6 ～ 10.7 nucleated cells/kg body weight of the recipient after thawing. The patients were conditioned with decreased dosage of immunosuppressive agents of CTX (60?mg/kg) and ALG (120?mg/kg). The engraftment state of the donor cells into recipients was confirmed by microsatellite DNA fingerprinting and fluorescent quantitative PCR analysis. Results Engrafted evidence has been found in 5 patients by molecular biology analyses showing donor-recipient mixed chimerism post transplant which was stable and persistent. After a median follow-up of 21 months (range 7～50), 4 patients were alive and disease free. One patient died of severe infection in the 3rd month from transplant, though the evidence of engraftment was obvious. Another patient also died in the early stage posttransplant of serious aspergillus infection without the engrafted proof.Conclusion Durable donor-recipient stable mixed chimerism can be achieved by unrelated UCBT in patients with SAA. Umbilical cord blood could be employed as a source of hematopoietic stem cell for adult transplantation.

Objective To explore the possibility of cytochrome P-450 3A5 (CYP3A5) genotype as a major factor to guide individualized employment of cyclosporin A(CsA) through a comparative study of CsA concentrations in the peripheral blood of hemopoietic stem cell transplant recipients with different CYP3A5 genotypes. Methods Olymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype CYP3A5 gene, and CsA concentrations were detected by a commercial fluorescence polarization immunoassay. Result There were significant differences in CsA concentrations, including standardized trough concentrations C0 and two h peak concentrations C2 , in the two CYP3A5 genotypes found in our samples, and both C0 and C2 in wild homozygotes were lower than heterozygotes. Conclusion CYP3A5 polymorphism is highly associated with CsA concentrations in hemopoietic stem cell transplant recipients, and CYP3A5 genotype may be a predictor to the dose requirement before clinically employment of CsA.

Objective To elucidate the effect of ex vivo expansion of umbilical cord blood (UCB) mononuclear cells (MNCs) on their implantation capability and the hematopoietic reconstitution, and to find a feasible way of applying ex vivo expanded UCB MNCs to clinical transplantation.Methods UCB MNCs were cultured in short-term in serum-free medium with different early acting cytokines combinations in order to observe the amplification result and cells apoptotic difference. The 6-day expanded cells were transplanted into sublethally irradiated NOD/SCID mice to assess the implantation and the hematopoietic reconstitution of the survival mice.Results UCB MNCs reached the best amplification result between day 6 and day 10 with the contribution of SCF, FL, IL-6 and IL-3 in common and the presence of Annxin V on the surface of cells obviously decreased. Six weeks after transplantation, CD45, CD34, CD33, CD3 and CD19 antigen could be detected by FCM on BM, spleen and thymus cells of the alive mice and the human specific Alu and Cart-Ⅰ repetitive sequence could be detected in the DNA obtained from peripheral blood by PCR.Conclusion After 6-day effective expanding with SCF+FL+IL-6+IL-3 UCB MNCs can be implanted into NOD/SCID mice successfully and contribute to reconstitute the multiple hematopoiesis.

Objective To explore clinical features of graft-versus-host disease (GVHD) after allogeneic non-myeloablative stem cell transplantation (NST) for haematologic diseases.Methods Eighteen patients were divided into three groups. Group A: Six severe aplastic anemia (SAA) adult patients underwent unrelated umbilical cord blood transplantation (UCBT). Group B: Combined transplantation of G-CSF primed allogeneic bone marrow and peripheral blood stem cells (PBSCs) was performed for 5 SAA patients. Group C: Seven malignant haematologic patients underwent transplantation of bone marrow cells for 3 patients or PBSCs for 4 patients. The protocol consisted of nonmyeloablative conditioning regimens based on anti-themocyte globulin (ATG) or anti-T-lymphocyte globulin (ALG). GVHD prophylaxis consisted of cyclosprine (CSA) and methylprednisolone (MP) for groups A and B, and methotrexate and CSA for group C. Mixed chimerism (MC) patients in group C were subjected to donor lymphocyte infusion (DLI).Results Four patients in group A achieved and sustained MC status, among them, one patient died of fungal septemia and one patient left hospital voluntarily. Three patients in group B achieved short period MC with donor chimerism more than 94 % at early stage post transplantation and converted into full donor chimerism (FDC) with long-term disease-free survival (DFS) and one patient developed chronic GVHD (cGVHD) 8 month post- trasplantation . Another two patients receiving donor stem cell infusion (DSI), one died of secondary mediastina lymphoma after 6 months and one patient recovered haematopoiesis. All patients achieved MC with haematologic partial remission (PR), and did not complicated acute GVHD (aGVHD) prior to DLI. Two cases died of severe infection and lost follow-up respectively. Another 5 patients gradually converted into FDC and achieved haematologic complete remission after 4, 3, 7, 5 and 4 DLIs, but they developed cGVHD (n=4), aGVHD (n=2) and myelosurppression (n=2).Conclusion The treatment of NST for SAA patients achieved better clinical effect with lower incidence of GVHD, and characterized by lower incidence of aGVHD and early mortality and higher incidence of cGVHD and infection for malignant haematologic diseases.

Objective To evaluate the efficacy and complications of nonmyeloablative stem cell transplantation(NST) for the treatment of nonmalignant haematologic diseases with high-risk rejection.Methods NST was performed for two patients with severe aplastic anemia(SAA) and one with beta-thalassemia major(TM).The protocal was designed on transplantation of granulocyte colony-stimulating factor(G-CSF) primed allogeneic bone marrow cells combined with perpheral blood stem cells(PBSCs) for two SAA patients,with conditioning regimen based on anti-lymphocyte globulin and reduced dose of cyclophosphamide(CTX).One TM patient was performed transplantation of PBSCs with conditioning regimen of anti-T-lymphocyte globulin(ATG),fludarabine and reduced dose of busulfan.Cyclosporin A combined with methylprednisone was used for graft-versus-host disease(GVHD) prophylaxis.Donor stem cells infusion(DSI) were underwent for three patients at 78,99 and 44 days post transplant respectively.Results Three patients achieved engraftment successfully with mixed chimera and the lowest white blood cell(WBC) of 0.26?10~9/L,0.5?10~9/L and 1.26?10~9/L respectively.The absolute neutrophil count achieved more than 0.5?10~9/L and platelet count achieved more than 20?10~9/L at days of 12d,3d,0d and 1d,5d,0d post transplant in three patients,respectively.The haematopoiesis and chimera were improved after DSI without complications of infection and GVHD in three patients.Conclusion The stem cells engraftment is achieved successfully with donor stem cell infusion followed NST for the treatment of nonmalignant haematologic disease patients with high-risk rejection.