OBJECTIVE To investigate the distribution and resistantce of three commonly encountered Gram-negative bacilli in our hospital,and provide doctors with the laboratory evidence of guiding antibiotic therapy.METHODS Bacterial identification and antibiotic susceptibility tests of isolates between Jan 2005 and Dec 2006 were performed by MicroScan WalkAway-40.RESULTS Totally 2024 common Gram-negative bacilli strains were isolated,807 of which were Escherichia coli,737 were Klebsiella pneumoniae,480 were Pseudomonas aeruginosa.The antimicrobial resistant rate of E.coli and K.pneumoniae to imipenem(2.2%) was lower than to the other antibiotics(2.7%).While 14.0% of P.aeruginosa isolates were resistant to imipenem.The antimicrobial resistant rate of all the isolated bacteria to piperacillin/tazobactam and amikacin was low,accounted less than 15%.CONCLUSIONS The resistant rate of three commonly encounted Gram-negative bacilli is increasing year by year.Therefore,monitoring bacterial drug resistance is very important for the rational use of antibiotics and the containment of multi-drug bacteria.

OBJECTIVE To investigate the in vitro antimicrobial susceptibility of Streptococcus pneumoniae and guide the rational use of drug clinically. METHODS We performed statistical analysis of the susceptibility of 62 strains of S.pneumoniae isolated in our hospital from May 2007 to Aug 2008. RESULTS S.pneumonia e could be isolated from various specimens,most of them were isolated from sputum(84.0%).S.pneumoniae could be detected from many hospital wards,but more strains isolated from pediatric(67.8%) and respiratory(11.4%) departments.A total of 67.7% of S.pneumoniae isolates were penicillin non-susceptible,the resistance prevalence to tetracycline was 87.1%,to erythromycin 79.0% and to trimethoprim/sulfamethoxazole 49%,while they were highly susceptible to ofloxacin(90.3%),vancomycin(91.9%),levofloxacin(95.2%),moxifloxacin(96.8%) and rifampicin(98.4%),and more strains showed multi-resistance from penicillin non-susceptible isolates. CONCLUSIONS The resistance to penicillin of S.pneumoniae is serious in our hospital.The tetracycline and erythromycin are not the best ckoice in treating S.pneumoniae infection,and the new fluroquinolones show strong activity against S.pneumoniae.

Objective To investigate the prevalence of qepA, quinolone efflux protein, among 41 unique clinical strains of K. pneumoniae producing ESBLs and to study the qepA-bearing isolates using the Diversilab system. Methods Identification and antimicrobial susceptibility were performed by Vitek-2 Compact System. Screening of qepA was carried out by PCR amplification. The NCBI BLAST program was utilized for sequence comparisons. qnr-bearing strains was evaluated by the Repetitive-sequence-based PCR(Rep-PCR) employing the Diversilab system. And the existence of rmtB was detected among these qepA contained isolates. Results qepA were detected in 5 isolates( 12.2% ). The Rep-PCR profiles produced by the Diversilab system showed that 2/5 of isolates were indistinguishable. And 60% of qepA-positive isolates were detected to harbor rmtB gene. Conclusion The data suggest the emergence of qepA-borne K. pneumoniae.

Objective To analyze the occurrence of traumatic carotid cavemons fistula (TCCF) resulted from the fracture of basilaris cranii, in order to find out the related factors to outcomes and to discuss the approaches to improving prognosis.Method Data of 312 patients with the fracture of skull base complicatcd with TCCF con-firmed angiography from 1999 to 2005 were analyzed. These patients were classified into patients with disable and patients without disabed. The factors potentially impacting on outcomes were analyzed. Results The overall inci-dence of TCCF in 312 patients with fracture of basilaris cranii was 3.8% .The incideucs of TCCF occurred in pa-tients with the fracture of anterior fossa, middle fossa and posterior fossa accounted for 2.4%, 8.3 % and 1.7 %, respectively. Between two cohorts of patients, there were no difference in age, gender, number of embelization proce-dares performed and the time from injury to appearence of the first symptom except the differencc in time from ap-pearence of the first symptom to the intravascular embohzation performed (P>0.05). Conclusions A relatively high incidence of TCCF occurs in patients with middle fossa fractures, especially those with transverse or oblique fractures. Prompt diagnosis and intervention can not be emphasized in case of patients with TCCF, and non inva-sive techniques for the early detection of TCCF under certain circumstance after brain or facial trauma should be considered so as to avoid a miss in the early diagnosis of middle fossa fracture to ensure favourable outcomes.

Objective To construct a stable infectious clone of S191 virus strain used for the pro-duction of live attenuated measles vaccine. Methods Full length cDNA of S191 strain and gene fragments encoding nucleocapsid(N),phosphoprotein(P)and RNA polymerase(L)were synthesis and respectively cloned into the vector pVAX1. The 293T cells were respectively transfected with the recombinant expression plasmids and co-cultured with Vero cells. The supernatants of cell culture were collected for identifying res-cued viruses. The indirect immunofluorescence assay was performed for virus identification. The rescued viruses at different passages in Vero cells and the sequences of the rescued viruses were analyzed. Results Restriction enzyme digestion and sequence analysis showed that the recombinant expression plasmids contai-ning the full length cDNA with an artificially engineered mutation at nucleotide 2101(C-A)and gene frag-ments encoding N,P and L proteins of S191 strain were constructed successfully. The N and P proteins were detected in Vero cells with immunofluorescence assay. A cytopathogenic effect on Vero cells was induced by rescued viruses. Conclusion The stable infectious clones of S191 virus used for the production of live at-tenuated measles vaccine were rescued successfully. An approach by using reverse genetics technique for S191 strain study was established which could be used for the development of new chimeric vaccines against measles virus.

Objective To investigate the clinical distribution and the drug resistance characteristics of Pseudomonas aeruginosa . Methods The results of the bacterial culture and the antimicrobial susceptibility test in the hospital from July 2007 to October 2008 were performed the retrospective analysis.Results Totally 335 strains of Pseudomonas aeruginosa were isolated,accounting for 9.2% of isolated pathogenic bacteria.The main specimen source was sputum,accounting for 77.6%.The ICU ward was the high incidence area.The resistance rates of Pseudomonas aeruginosa to amikacin,piperacillin/tazobactam,tobramycin,levofloxacin, cefepime,gentamicin,ticarcillin and ciprofloxacin were less than 10%.The resistance rates of imipenem-insensitive Pseudomonas aeruginosa to aztreonam,ceftazidime,ciprofloxacin,levofloxacin,piperacillin/tazobzctam were significantly higher than those in imi-penem-sensitive Pseudomonas aeruginosa (P <0.05).Conclusion The multiple drug resistance phenomena of Pseudomonas aerugi-nosa generally exist,amikacin,piperacillin/tazobactam and tobramycin are recommended for the treatment of infections caused by Pseudomonas aeruginosa .

Objective To construct a mutant strain of Streptococcus mutans ( S.mutans ) with clpC-deletion and to investigate the role of clpC gene in genetic competence.Methods The fragment of clpC gene and the kanamycin resistant cassette flanked by two loxP sites were amplified by PCR.The purified fragment of clpC gene was cloned into pMD-19T simple vector to construct pCKX1.The pCKX1 vector was digested with ClaⅠ/EcoRⅠ, then blunted and introduced into lox71-KMR-lox66 to obtain pCKX2 vector via homologous recombination.The pCKX2 vector was linearized with SalⅠ and transformed into S.mutans UA159 strain.The positive strains constructed via homologous recombination were screened with kanamycin and transformed with the thermosensitive plasmid pCrePA.The KMR cassette was excised after incubating at 30℃ for 48 hours.Then the pCrePA plasmid was removed after overnight incubating at 37℃for the prepara-tion of clpC-deletion mutant.Total RNA were extracted from the S.mutans UA159 strain and the clpC-dele-tion mutant strain respectively, and then reverse transcribed into first strand cDNA.The target gene frag-ments were amplified by RT-PCR and analyzed by the agarose gel electrophoresis and sequencing.After be-ing verified by PCR and sequencing, the S.mutans UA159 strain and the clpC-deletion mutant strain were re-spectively transformed with E.coli-S.mutans shuttle vector pDL276 to observe the competence development induced by the competence-stimulating peptide (CSP).Results The PCR and sequencing results showed that the pCKX2 vector and the mutant strain with clpC-deletion were constructed successfully via homologous recombination.No clpC gene was detected in the clpC-deletion mutant as indicated by RT-PCR analysis.The formation of competent clpC-deletion mutant was delayed and the competence state was prolonged as com-pared with its parent strains.Conclusion The clpC gene negatively regulated the formation of competent S.mutans.

Objective To investigate the genotype and epidemiology of plasmid‐mediated AmpC β‐lactamases produced by the clinical strains of Escherichia coli and Klebsiella pneumoniae .Methods A total of 176 clinical nonrepetitive cefoxitin non‐sensitivity isolates of Escherichia coli and Klebsiella pneumoniae was collected from July 2011 to August 2012 .Polymerase chain reaction (PCR) for AmpC enzyme gene amplification and DNA sequencing were carried out for genotype of AmpC beta‐lactamases .Results The results of PCR showed that the positive rate of ampC of the 176 strains of Escherichia coli and Klebsiella pneumoniae AmpC was 18 .2% ,mainly DHA type ,counting for 59 .4% ,CIT counting for 37 .5% ,EBC counting for 3 .1% .The positive rate of ampC of Escherichia coli was 11 .4% ,mainly CIT type ,counting for 77 .8% ,the positive rates of DHA type and EBC type both were 11 .1% .The positive rate of ampC of Klebsiella pneumoniae were 23 .7% ,mainly DHA type ,counting for 78 .3% ,CIT type count‐ing for 21 .7% .The results of DNA sequencing showed that there were 18 strains DHA‐1 type and 1 strain ampC gene type of Morganella morganii in DHA type strains ,the concordance rate was 97 .0% ,10 CIT type strains was CMY‐2 type ,1 strain was CMY‐42 ,one strain was CMY‐4 type ,EBC type was ampC gene type of Enterobacter cloacae ,the concordance rate was 99 .0% .A total of 32 strains of gene sequencing were registered as KJ127248 - KJ127279 in GenBank .Conclusion The main genotypes of plasmid‐mediated ampC enzyme produced by Escherichia coli and Klebsiella pneumoniae were CMY‐2 and DHA‐1 respectively .

Objective To study a rehabilitation nursing menus for neurogenic large intestine dysfunction.Methods Various nursing approaches were used for defecation dysfunction.Results and Conclusion 94.60% patients improved in the second week,which including gained more awareness,control,and spend less time of defecation.

Primary testicular lymphoma comprises 1% to 9% of testicular neoplasms and represents 1% to 2% of all non-Hodg-kin lymphomas. Histologically, the majority of the tumor consists of diffuse large B-cell non-Hodgkin lymphomas that are of intermedi-ate- or high-grade neoplasm. Clinically, the disease typically presents as a painless testicular swelling that develops over a span of weeks to months. B symptoms such as fever, weight loss, and anorexia are present in 25% to 41% of the patients. This tumor is an ag-gressive type, with frequent invasion of the epididymis, spermatic cord, and scrotum, as well as a marked tendency to relapse, especial-ly in the CNS. The treatment is mainly based on orchiectomy (mostly in stages ⅠE and ⅡE) regardless of its association with prophy-lactic irradiation of the scrotum and administration of intrathecal chemotherapy, cyclophosphamide, doxorubicin, vincristine, and pred-nisone regimen chemotherapy plus rituximab (R-CHOP) (stages ⅢE and ⅣE) and radiotherapy. The multi-modality treatment marked-ly improved progression-free and overall survival. We introduce as reference one case that received a multidisciplinary comprehensive discussion in the Department Lymphoma, Tianjin Medical University Cancer Hospital.