OBJECTIVE To investigate the distribution of pathogens and their antibiotic resistance in lower respiratory tract infection(LRTI) from intensive care unit(ICU) in our hospital,and provide basis for rational selection of clinical drugs.METHODS Pathogens were detected from qualified sputum specimens in LRTI from ICU and identified by VITEK-AMS60 automatic microbial analyzing system.Drug susceptibility was determined by KB test.RESULTS From 320 sputum specimens 367 pathogens were detected between from Jan 2007 to Mar 2008,including 261 strains(71.1%) of Gram-negative bacilli,70 strains(19.1%) of fungi,and 36 strains(9.8%) of Gram-positive cocci.21.8% Of the isolated pathogens were Acinetobacter baumannii,with 16.7% of drug-resistant rate to cefoperazone/sulbactam and over 71% to other 13 antibiotic agents.The rate of extended spectrum ?-lactamases(ESBLs) producing Escherichia coli isolates and Klebsiella pneumoniae ones were 65.2% and 72.0%,respectively,comparing to 84.6% for meticillin-resistant Staphylococcus aureus(MRSA).CONCLUSIONS Gram-negative bacilli are the major pathogens of LRTI in ICU,in which A.baumannii shows with a high rate of drug-resistance,followed by fungi,which should attract the clinician′s more attention.

Objective To assess the application value of REP-PCR in genotyping of candida glabrata strains in clinical pratice. MethodsFrom 2009 to 2010, thirty-eight candida glabrata strains were isolated from Shanghai Ruijin Hospitals, Shanghai Renji Hospital, Shanghai Huashan Hospital, Anhui Medical University Hospital, Shenzhen People's Hospital. Six loci in housekeeping genes (FKS, LEU2,NMT1, TRP1, UGP1 and URA3 ) were amplified and sequenced. The sequences were compared with the MIST database and allele profile and sequence type (ST) were obtained. With primers Ca21, Ca22 and Com21 used to amplify the adjacent variable gene regions,the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Finally, the results of these two genotyping methods were compared. ResultsFor REP-PCR, Ca22-Com21 has the best genotyping effect. REP-PCR and MLST have the same genotyping results. Five REP-PCR types were found in 38 candida glabrsta isolates. Type A,B, C, D and E strains from REP-PCR were genotyped as ST 7, 3, 19, 45 and new type respectively byMIST. REP-PCR saves time compared with MIST. Conclusions REP-PCR offers a simple and rapid method for molecular typing, with similar discriminatory power with MIST. Therefore, REP-PCR can be the preferred choice in laboratory, especially for a large number of isolates.

Objective:To study the effects of smokeless tobacco extract(ST)on the proliferation and the heat shock protein 70 (HSP70)expression of human periodontal ligament cells(hPDLCs).Methods:hPDLCs were cultured in vitro and identified by im-munohistochemistry(IHC).The cells were stimulated with ST at 0.01 6 -50 g/L respectively for 24 h,the proliferation was examine by MTT assay,HSP70 expression was detected by immunehistochemical staining and Western Blot.Results:ST inhibited the prolifer-ation and increased HSP70 expression in cytoplasm and nucleus at 0.4 -50 g/L dose dependantly.Conclusion:ST may inhibit the proliferation and increase HSP70 expression of hPDLCs in a dose depandant manner.

Objective To observe the effects of adiponectin on mRNA and protein expressions of connective tissue growth factor (CTGF) in hepatic stellate cells (HSCs) and the levels of procollagen type Ⅲ (PC Ⅲ) and hyaluronic acid (HA).Methods Cultured rat HSCs were treated with different concentrations of adiponectin.CTGF mRNA and protein expressions were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western Blot.The levels of PCⅢ and HA were detected by enzymelinked immunosorbent assay (ELISA).Results Compared with the control group,the result of RT-PCR showed that the four groups had different degrees of inhibitory effect,of which group D exhibited the strongest inhibitory effect.The absorbance ratio was 1.54 ±0.18,1.21 ±0.14,0.96 ±0.10,and 0.79 ± 0.08,respectively (t =2.42,P ＜0.05;t =2.73,P ＜0.05;t =3.28,P ＜0.01;t =4.67,P ＜0.01).Western Blot also indicated that four groups had different degrees of inhibitory effect,of which D group exhibited the strongest inhibitory effect.The ratio of integral absorption was 1.54 ± 0.18,1.21 ±0.14,0.96±0.10,and 0.79 ±0.08,respectively (t =2.84,P ＜0.01;t =4.05,P ＜0.01;t =6.25,P ＜0.01;t =9.72,P ＜0.01).The levels of PCⅢ and HA secreted in culture media were also decreased.It was significantly decreased with the concentration of adiponectin increased.Conclusion Adiponectin can inhibit the levels of PC Ⅲ and HA,which may be achieved through reducing CTGF mRNA and protein expressions.

Objective To summarize the radiography ,CT and MRI findings of maxillary arteriovenous malformation (AVM ) . Methods Seventeen patients with maxillary AVM underwent panoramic radiography (3 patients) and enhanced MRI examination (6 patients) ,all the patient underwent enhanced CT examination .The clinical manifestation and imaging findings of maxillary AVM ,in‐cluding the location ,shape ,margin ,inner texture ,involvement of adjacent structures ,the density and signal intensity of the lesions , were analyzed .Results The lesions mainly located in molar areas (15/17) .The major clinical manifestation were repeated bleeding and acute bleeding .Other symptoms included swelling of the face ,pulsatile soft mass and anesthesia .The panoramic radiography only showed increased density .According to the change of the maxilla on enhanced CT images ,the lesions could be devided into two types :type Ⅰ ,showed intraosseous osteolytic destruction and cortical expansion (n=12);type Ⅱ ,showed“ground glass”appearance (n=5) .Bone destruction and soft tissue involvement ,widened feeding artery and draining veins ,elevated maxillary sinus were shown in all patients .External jugular vein enlargement and early enhancement (n=14) and root resorption (n= 6) were also detected . Flowing void on T1 and T2 weighted images (type Ⅰ) and hypo‐or isointense on T1WI ,hyperintense on T2WI and obvious enhance‐ment after injecting contrast material (type Ⅱ ) could observed .Conclusion Enhanced CT examination could be primarily recom‐mended and observe the extent of lesions and situation of vessels invaded of maxillary AVM .

Background Diabetic retinopathy (DR) is a chronic inflammatory disease,with pathological changes of retinal microvessels.Studies demonstrated that sericin has anti-inflammation and anti-oxidation effects,inferring that sericin might play a protective effect on diabetic microangiopathy.Objective This study was to investigate the effects of sericin on intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1) and Cx43 expressions in retina and explore the protection of sericin to retinal microangiopathy in diabetic rats.Methods Forty-eight specific pathogen free male SD rats were randomly divided into normal control group,diabetic model group,sericin-treated group and calcium dobesilate-treated group,with 12 rats for each group.The diabetic models were established by intraperitoneal injection of streptozotocin (STZ) for 3 consecutive days and feeding up with high lipid foods.Normal saline solution,2.4 g/(kg · d) sericin solution and 0.2 g/(kg · d) calcium dobesilate were used by gavage administration in the rats of the diabetic model group,sericin-treated group and calcium dobesilate-treated for 3 months group,respectively.The rats were sacrificed and retinal sections were prepared,and the retinal morphology was examined by hematoxylin-eosin staining.The expressions of ICAM-1,VCAM-1 and Cx43 proteins and mRNA in retinas were detected by Western blot assay and reverse transcription PCR.The use and care of the rats complied with Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission and ARVO statement.Results The retinal structure was normal in the normal control group.The swell and rupture of inter limiting membrane (ILM),scatter vascular endothelial cell nuclei breakthrough ILM and decrease of retinal ganglion cells (RGCs) were displayed in the diabetic model group;while in the sericin-treated group and calcium dobesilate-treated group,the mild thickening of ILM and disorder of retinal cells were obtained.The relative expression levels of ICAM-1 and VCAM-1 were significantly raised and those of Cx43 were reduced in the diabetic model group,sericin-treated group and calcium dobesilate-treated group when compared with the normal control group (all at P＜0.05).Compared with the diabetic model group,the expressions of ICAM-1 and VCAM-1 proteins and mRNA in the sericin-treated group were significantly reduced (ICAM-1 protein:0.834 3±0.032 1 vs.0.918 9±0.042 4;VCAM-Ⅰ protein:0.726 4±0.011 2 vs.1.235 0±0.078 9;ICAM-1 mRNA:0.716 3±0.008 6 vs.0.956 8±0.012 5;VCAM-1 mRNA:0.393 7±0.035 0 vs.0.477 9±0.020 6) and those of Cx43 protein and mRNA were evidently elevated (Cx43 protein:0.133 1 ±0.015 3 vs.0.039 2±0.002 0;Cx43 mRNA:0.676 8 ±0.064 8 vs.0.430 8±0.111 3) (all at P＜0.05).Conclusions Sericin can relieve retinal microangiopathy and protect retina against the pathogenesis and development of DR by down-regulating the expressions of ICAM-1,VCAM-1 and upregulating the expression of Cx43 in retinas of diabetic rats.

Objective To investigate the factors influencing the prevalence of echinococciasis in Gansu Province.Methods With the method of stratified random sampling,all the administrative villages in pastoral areas,half pastoral areas,agriculture and urban areas in the 72 counties in Gansu Province were selected;according to the layers of the population of the county population proportion calculated,the layer investigation numbers were determined,and a total of 16 administrative villages were selected in each county.In the 16 administrative villages,more than 200 people of permanent residents were surveyed in each village (shortage was made up from a nearby village),at least 3 200 people were investigated in a county.By using the combined method of inquiry and field observation,natural factors,such as drinking water,farming and animal husbandry production data were surveyed.All data were analyzed using SPSS 19.0 statistical software.Unauy linear and multiple linear regression analysis of influencing factors and the relationship between the prevalence were analyzed.Results Unary linear regression analysis showed that longitudes,latitudes,altitudes,average annual temperature,annual rainfalls,secondary industries,animal husbandry towns,drinking water sources,the number of epidemic towns,epidemic villages,per capita net incomes in animal husbandry regions,the total number of livestock and the number of sheep were factors influencing the prevalence of echinococciasis (F =4.705,P ＜0.05).Latitude,altitude,animal husbandry towns,the number of epidemic towns,epidemic villages,per capita net incomes in animal husbandry regions,the total number of livestock and the number of sheep and the prevalence of echinococciasis was positively correlated (r =0.282,0.285,0.387,0.508,0.540,0.317,0.475,0.594,all P ＜0.05);longitude,average annual temperature,annual rainfall,secondary industries,drinking water sources and the prevalence of echinococciasis was negatively correlated (r =-0.311,-0.244,-0.244,-0.389,-0.311,all P ＜0.05).Multiple linear regression analysis showed that differences of interactions of annual rainfalls,drinking water sources,epidemic villages,per capita net incomes of herdsmen,secondary industries and the total number of livestock between groups were statistically significant (t =-1.822,-3.920,3.013,1.715,-1.609,3.264,all P ＜0.05).Conclusion The factors influencing the prevalence of echinococciasis in Gansu Province are correlated with annual rainfalls,drinking water sources,epidemic villages,per capita net incomes of herdsmen,secondary industries and the total number of livestock.

OBJECTIVE To develop a real-time fluorescence PCR assay to detect the genes encoding thermolabile(hemolysin)(TLH),thermostable direct hemolysin(TDH) and TDH-related hemolysin(TRH) of Vibrio(parahaemolyticus).METHODS The genes of TDH and TRH were selected as target ones of thermostable direct and TDH-related hemolysin,and TLH gene as a specific genomic marker for V.parahaemolyticus.Designed and synthesized the primers and Taqman probes,we investigated 487 stool samples of doubt foodborne illness patients by real-time fluorescence PCR.RESULTS The sensitivity of the assay for TLH and TDH was 1.0?10~2copies,but the sensitivity of TRH was 1.0?10~3copies. Among 487 samples,112 V.parahaemolyticus strains were found;101 samples of these strains showed the production of TDH;none of them was positive for TRH.CONCLUSIONS The Taqman PCR is a rapid and sensitive method to detect the TLH,TDH and TRH of V.parahaemolyticus,it is well suited for screening large numbers of samples at the same time;and TDH is one of the primary virulence factors in clinical isolated V. parahaemolyticus.

Objective To investigate whether the amount of blood transfusion affects the lengths of stay (LOS),costs,and outcomes of hospital patients or not,and to prepare for the execution of patient blood management.Methods The data of hospital patients,who had been administrated with blood in our hospital during 2016,were collected.And the influence of blood transfusion volume on LOS,costs and outcomes of patients was analyzed retrospectively.Results LOS,costs and outcomes of patients vary significantly with the amount of blood transfusion (P＜0.01).There were positive correlations between the total amount of blood transfusion and LOS,costs,and outcomes of patients.The Spearman correlation coefficient was 0.317,0.497,0.290,respectively (P＜0.01).Plasma preparation transfusion volume has a great influence on LOS,costs,and outcomes than red blood cell (P＜0.05).The transfusion volume of death patients was significantly higher than that of the survival (P＜0.01).In particular,the amount of transfused plasma and precipitation was distinctly higher than that in death patients(P＜0.01).Conclusion Blood transfusion volume affects LOS,costs and outcomes of hospital patients.The administration of plasma preparations should deserve more attention.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of TLR4 and its role in lipopolysaccharide (LPS)-induced NF-κB activation and CD40 expression in rat peritoneal mesothelial cells (RPMCs). Methods RPMCs were harvested from Spragne-Dawley rat peritoneal cavity and maintained under defined in vitro condition. The cells were treated with Ang Ⅱ at different concentrations (10-9, 10-8, 10-7, 10-6 mol/L) and exposed to Ang Ⅱ (10-7 mol/L) for different times (1, 2, 4, 8, 12, 24, 48 h for mRNA and 6, 12, 24, 36, 48 h for protein, respectively). Meanwhile, the influence of AT1 receptor antagonist (AT1R, losartan, 10-5 mol/L) and AT2 receptor blocker (AT2R, PD123177, 10-5 mol/L) on the TLR4 induced by Ang Ⅱ was observed. After synchronization for 24 hours, the cells were randomly assigned to four groups: the control group, the Ang Ⅱ (10-7 tool/L) group, the LPS (1 mg/L) group, the Ang Ⅱ (10-7 mol/L) plus LPS (1 mg/L) group, which were used to investigate the effects of Ang Ⅱ on the NF-κB activation and CD40 expression induced by LPS. The mRNA expression of TLR4 and CD40 was measured by RT-PCR and the protein abundance of TLR4, NF-κB p65, phospho-p65, IKBα and phospho-IκBα were analyzed by Western blot. Immunofluorescence was performed to determine the subcellular localization of p65 subunit of NF-κB. Results (1) Treatment of RPMCs with Ang Ⅱ resulted in a concentration-dependent increase in the expression of TLR4. Ang Ⅱ at 10-9, 10-8, 10-7 and 10-6 mol/L increased TLR4 mRNA expression by 70.5%, 89.5%, 102.9%, and 121.9%, respectively and protein expression by 12.1%, 27.7%, 51.2%, and 41.6%, respectively (P<0.01). Treatment of RPMCs with 10-7 mol/L Ang Ⅱ resulted in a time-dependent increase in the expression of TLR4, with the peak of mRNA expression at 8 and 12 h (P<0.01) and the protein expression at 12 and 24 h (P<0.01). (2) Losartan antagonized Ang Ⅱ-stimulated expression of TLR4 by 33.5% (P<0.05), PD123177 had no such effect (P0.05). (3) Treatment of RPMCs with LPS (1 mg/L) for 60 rain significantly increased the ratio of phospho-IκBα to IκBα by 362.6% (P< 0.01) , phospho-p65 to p65 by 67.4% (P<0.05), and LPS (1 mg/L) for 4 h significantly increased the expression of CD40 mRNA by 299.9% (P<0.01) compared to the control group. In comparison to the LPS (1 mg/L) group, preincubation of RPMCs with AngⅡ (10-7 mol/L) for 24 h then treated with LPS (1 mg/L) for 60 rain significantly increased the ratio of phospho-IκBα to IκBα by 49.1% (P<0.01), phospho-p65 to p65 by 29.3%(P<0.05), and LPS (1 mg/L) for 4 h significantly increased the expression of CD40 mRNA by 56.8%(P<0.01). (4) The p65 subunit of NF-κB was dominantly distributed in the cytoplasm in the control and Ang Ⅱ group. Following exposure to LPS for 60 min, p65 subunit labeling was upregulated and translocated into the nuclei. A significantly increased nuclear staining of p65 in ceils treated with Ang Ⅱ plus LPS were observed. Conclusions Ang Ⅱ induces the expression of TLR4 in dose- and time-dependent manner in RPMCs, resulting in enhanced NF-κB signaling and induction of CD40 expression, Locally produced Ang Ⅱ in the peritoneum may play an amplified role in LPS-induced peritoneal inflammation.