OBJECTIVE To investigate spectrum and drug resistance of No-Pseudomonas aeruginosa,to provide the basis for proper use of antibiotics and prevent patients from hospital acquired infection.METHODS API staph analysis system and the K-B agar diffusion test according to NCCLS were used to identify 357 strains of No-Pseudomonas aeruginosa and to performe drug sensitivity test.RESULTS From 2004 to 2008,357 strains of No-Pseudomonas aeruginosa were isolated.There are 233 Stenotrophomonas maltophilia strains(65.27%,233/357),55 Burkholderia cepacia strains(15.41%,55/357).Sputum was the main sourse of No-P.aeruginosa(88.52%,316/357).The isolation rate of No-P.aeruginosa was the highest in intensive care units(38.10%,136/357),the next was department of respiratorymedicine(23.53%,84/357).The results of drug susceptibility test showed that the resistance rate of No-P.aeruginosa to most drugs were higher,only the resistance rates of S.maltophilia to minocycline,gatifloxacin and levofloxacin were lower,(3.26%,20.55% and 20.83% respectively),The resistance rates of B.cepacia to meropenem,piperacillin/tazobactam and Imipenem were lower,being 29.17%,33.33% and 39.62% respectively.CONCLUSIONS The most common strains is S.maltophilia and Burkholderia cepacia,the most common site of infection is lower respiratory tract,Most strains of S.maltophilia are isolated from ICU and department of respiratory diseases.The result of drug sensitive test showed that there were high rates of multiple drug resistance in No-P.aeruginosa and treatment of No-P.aeruginosa infection should be based on the results of the drug susceptibility test.

Objecfive To investigate the effects of treatment for cerebrovascular disorder patients with heparin and low molecular weight heparin(LMWH) on serum PAPP-A concentrations and provide the basis for evaluating the clinical significance of PAPP-A in the following study.Methods Forty cases with cerebrovascular disease from Qilu Hospital from November 2009 to May 2010 were collected in this study.Blood samples were taken before and after drug administration.All cases were divided into four groups according to situation of medication.Group A consisted of 10 patients who received subcutaneous LMWH anticoagulation therapy, and blood samples were collected before LMWH injection, three hours after subcutaneous LMWH anticoagulation therapy in the first day, the second day and the seventh day and 24 hours after the last injection. Group B consisted of 10 patients who did not receive LMWH therapy, and blood samples were collected immediately after admission, the first day, the second day and the seventh day after admission. Group C consisted of 10 patients with percutaneous carotid intervention who received intravenous heparin at the beginning of stenting, and blood samples were collected from the arterial sheath just before angiography and heparin administration, and at 3, 5, 15, 40 and 100 min after heparin administration. Group D consisted of 10 patients who received carotid angiography but LMWH-free therapy,and blood samples were collected from the arterial sheath just before and after angiography. Serum PAPP-A concentrations were analyzed by ELISA to evaluate the differences of intra-groups and differences at different time points of inter-groups. Results In group A, PAPP-A concentrations were time dependent and elevated gradually from 12. 36 (9. 90-14. 32) mIU/L before LMWH injection to 21.80 (23.50-19.73) mIU/L at the seventh day after injection (M=38. 72, P < 0.01 ). In group C, there was a rapid increase of PAPP-A concentration from 12. 86 ( 9. 67-14. 05 ) mIU/L to 51.56 ( 44. 20-66. 00 ) mIU/L within 5 min after intravenous heparin injection (M=46. 06, P <0. 01 ). The PAPP-A concentration of one week after LMWH administration in group A was 21.80 (23.50-19.73) mIU/L, significantly higher than that in group B [11.81 (9. 21-12. 89) mIU/L] (U<0. O01, P<0.01). The PAPP-A concentration at 15 min after heparin administration in group C was 43.70 (37.70-54. 30) mIU/L, significantly higher than that after angiography in group D [14. 18 (11.25-15. 86) mIU/L] ( U<0. 001, P <0. 01 ). The peak level of blood PAPP-A after subcutaneous LMWH injection was significantly lower than that after intravenous heparin injection. The concentrations in group A and C were 21.80 ( 23.50-19. 73 ) and 51.56 (44. 20-66. 00) mIU/L respectively, and had a significant difference ( U=0. 999, P < 0. 01 ) . Conclusions Both intravenous heparin and subcutaneous LMWH administration induce an increase in serum PAPP-A concentration. The effect of drug should be considered when PAPP-A is selected as an evaluation indicator.

Objective To observe the effects of sodium‐selenite on human hypertrophic scar fibroblasts proliferation in vitro . Methods Human hypertrophic scar fibroblast culture was conducted in vitro ,the status of fibroblast proliferation of the 4th gener‐ation cells was tested by CCK‐8 ,which was divided into experimental group and control group ,the experimental group was divided into six groups (A ,B ,C ,D ,E ,F) ,and were added an equal volume‐containing 2 .5 ,5 .0 ,10 .0 ,20 .0 ,40 .0 ,80 .0 μmol/L concentra‐tions of sodium selenite in 10% FBS culture medium ;the control group added an equal volume of 10% FBS culture medium ,testing cell proliferation by CCK‐8 at 24 ,48 ,72 ,96 h respectively ;testing different concentrations of sodium‐selenite cell survival situation after 24 h by Live/dead reagent ;immunohistochemical was used to test intracellularⅠ ,Ⅲ type collagen expression after 24 h .Re‐sults (1)With the increased of concentration ,the inhibition rate of fibroblasts gradually increased as the concentration of sodium‐selenite ranged in 2 .5-80 .0 μmol/L(P<0 .05);(2) the inhibition rate of sodium‐selenite on fibroblasts gradually increased at the same concentration with time(P<0 .05);(3)Live/dead reagent test results showed that apoptosis cell number increased with the concentration increasing ;(4 ) With concentrations of sodium‐selenite increasing ,typeⅠ ,Ⅲ collagen expression of fibroblast de‐creased gradually .Conclusion Sodium‐selenite can inhibit human hypertrophic scar fibroblast proliferate in vitro and reduce Ⅰ ,Ⅲcollagen expression of fibroblast type.

Objective To explore the application value of plasma sHLA-G in diagnosis of CIN and cervical cancer. Methods The plasma sHLA-G levels were detected by ELISA in 102 cases with cervical cancer( FIGO Ⅰ stage 32 cases, Ⅱ stage 28 cases, Ⅲ stage 25 cases and Ⅳstage 17 cases; tumor size:＜4 cm 63 cases and ≥4 cm 39 cases; squamous cell carcinoma 78 cases and adenocarcinoma 24 cases;cell differentiation:well 57 cases, moderate 29 cases and poor 16 cases; lymph nodes metastasis negative64 cases and positive 38 cases ), 72 cases with CIN( Ⅰ grade 21 cases, Ⅱ grade 25 cases and Ⅲ grade26 cases ) and 20 cases of healthy controls. The diagnostic value of sHLA-G and its correlations with clinical parameters were analyzed. Results The plasma levels of sHLA-G were 193.6( 151.3-287.4 ) kU/L in cervical cancer group, 48.3( 34.6-57.2 ) kU/L in CIN Ⅰ group, 91.3( 68.2-118.6 ) kU/L in CIN Ⅱ group, 106.4( 73.8-165.7 ) kU/L in CIN Ⅲ group and 45.2( 38.0-55.5 ) kU/L in health control group.The level of sHLA-G was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and healthy control group( U value of 8.832, 6.456, 4.017, 9.873, P ＜ 0.05,respectively ). The level of sHLA-G was significantly higher in CIN Ⅱ group and CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 4.361,4.892, 5.139, 5.485, P ＜0.05, respectively ).The levels of SCC Ag in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 0.43( 0.38-0.69 )μg/L, 0.47( 0.35-0.72 )μg/L, 0.65( 0.53-0.81 )μg/L, 0.82( 0.54-1.03 )μg/L and 1.02( 0.62-1.87 )μg/L. The level of SCC-Ag was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.926, 4.877, 8.132,P ＜0.05, respectively ). The level of SCC-Ag was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 6.574, 6.763, P ＜0.05, respectively ). The levels of CA125 in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 14.38 ( 6.14-21.82 ) kU/L, 15.42( 6.25-23.53 ) kU/L, 21.34( 9.82-32.58 ) kU/L, 25.69( 14.47-38.71 )kU/L and 27.72( 14.29-43.87 ) kU/L. The level of CA125 was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.564, 4.522, 7.429, P ＜0.05, respectively ). The level of CA125 was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 5.871, 5.435, P ＜0.05, respectively ). ROC curve analysis showed AUC for sHLA-G was 0.828( 95% CI:0.768-0.879 ), which was high as compared with the AUC of SCC-Ag [ 0.727( 95% CI:0.658-0.788 );Z = 2.294, P ＜ 0.05 ] and the AUC of CA125 [ 0.705( 95% CI:0.636-0.769 );Z =2.842 ,P ＜0.05 ]. There was no significant difference of diagnostic efficiency between SCC and CA125( Z =0.672, P ＞ 0.05 ). When cutoff value of sHLA-G was 109.6 kU/L, the diagnostic sensitivity,specificity, positive predictive value, negative predictive value and accuracy rate were 86.3%, 76.1%,80.0%, 83.3%, and 78.4%, respectively. The levels of sHLA-G in cervical cancer patients were significantly correlated with FIGO stages and lymphoid node metastasis ( U value of 6.085, 4.451, P ＜0.05, respectively ), while there were no significant differences between the levels of sHLA-G and age,tumor size, histological type and cell differentiation( U value of 1.274, 1.956, 1.268, 2.719, P ＞0.05,respectively ). Conclusions sHLA-G can be used for the early screening of cervical cancer and its precancerous lesion. It could also be used as an index for judging progression and lymphoid node metastasis.

Objective To establish a direct reverse transcription real-time fluorescence quantitative polymerase chain reaction ( RT-qPCR-D ) method for detecting serum circulating B cell-specific moloney murine leukemia virus integration site-1 (Bmi-1) mRNA, and analyze the levels of serum circulating Bmi-1 mRNA in colorectal cancer patients by using of this method for exploring its diagnosis value in colorectal cancer.Methods Methodology establishment.RNA was extracted from colorectal cancer HT 29 cell line, and detection standard curves of Bmi-1, ubiquitin C ( UBC), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) mRNAs were established , then the amplification efficiencies were calculated.Bmi-1 mRNA level was directly detected in serum and preparation buffer mixture , then the specificity of assay was evaluated by melting curve, and detection limit was observed through diluted serum samples.The serum circulating Bmi-1 mRNA levels were detected by ELISA in 158 cases with colorectal cancer , of which there were 26 cases of tumor node metastasis ( TNM)Ⅰstage, 53 cases of TNMⅡ, 47 cases of TNMⅢ, 32 cases of TNMⅣand 53 cases of controls with normal colonoscopy collected from January 2008 to January 2009 in Qilu Hospital of Shandong University.Comparisons of groups were determined by applying Mann-Whitney U test or Kruskal-Wallis test, and receiver operating characteristic ( ROC) curves were established to illustrate the diagnostic performance.Results The log values of Bmi-1, UBC and GAPDH showed good linear correlations with quantification cycle (Cq) values(R2 =0.990, 0.990, 0.991, all P <0.001), and the amplification efficiencies were 0.875, 0.917 and 0.935, respectively.Using the established RT-qPCR-D method, the peak of melting curve of Bmi-1, UBC and GAPDH mRNAs were single, the detection limit was up to 1.25μl.The levels of serum circulating Bmi-1 mRNA detected by RT-qPCR-D were 0.138 ( 0.078-0.228 ) in colorectal cancer stage Ⅰ patients, 0.163(0.067 -0.287) instage Ⅱ patients, 0.217(0.072-0.267) instage Ⅲpatients, 0.273(0.139 -0.419) in stage Ⅳ patients and 0.021(0.008 -0.029) in health controls, a significant difference was found among groups ( H =89.5, P <0.001 ).The levels of serum circulating Bmi-1 mRNA in each stage colorectal cancer were all significantly higher than that in control group(U=58.0, 287, 246, 72.5,all P<0.001).The levels in Ⅳstage patients were significantly higher than those in other stages patients (U=247, 590, 540,P=0.008, 0.020, 0.035), while no significant differences among Ⅰstage,Ⅱstage and Ⅲstage patients(U=633, 514, 1170,all P>0.05).ROC curve analysis showed area under the ROC curve ( AUC) for serum circulating Bmi-1 mRNA was 0.921(95%CI=0.876-0.953), which was significantly superior to the AUC of CEA (0.745, 95%CI=0.680-0.802, Z=4.697, P<0.001 ).When cutoff value was 0.034, the diagnostic sensitivity and specificity was 89.2%(141/158) and 90.6%(48/53), while 41.8%(66/158) and 73.6%(39/53) using CEA.The AUC for combination of circulating Bmi-1 mRNA and CEA was 0.933(95%CI=0.890-0.963 ) , which was no statistical significance when compared with the AUC of circulating Bmi-1 mRNA(Z=4.697, P>0.05).Conclusions The study establishes a higher sensitive, specific for detecting serum circulating Bmi-1 mRNA. Based on this method , serum circulating Bmi-1 mRNA is found to be increased in colorectal cancer , and is superior to traditional tumor marker CEA in diagnosis of colorectal cancer, which may become a potential detection index for early detection of colorectal cancer.