Objective To investigate the present situation and reasons of emergency department overcrowding, and put forward effective mitigation strategy. Methods By using input-throughput-outputmodel and choosing the method of fishbone diagram analysis, detaily analysis factors of emergency department overcrowding, then implement three- dimensional intervention in hospital, department, personal. Results Patients who stay>48 h in the emergency department(ED) decreased from 11.9％(15225/127941) to 5.3％(7245/136698); patients who stay > 6 h in the Frist Aid Room of ED decreased from 54.6％(3016/5526)to 17.8％(987/5526), both have statistically significant (χ2=3705.04, χ2=1186.32, P<0.01);before the intervention the National ED Overcrowding Study (NEDOCS) index at 0900, 1300, 2100, 0100 was 234.22 ± 62.31, 253.55 ± 59.26, 303.73 ± 160.24, 187.36 ± 25.73; after the intervention the degree of crowdedness of ED at this four points of time was significantly alleviated, NEDOCS index was 193.09 ± 31.87, 187.09 ± 22.65, 187.36 ± 25.73, 154.03 ± 21.56;there were significant differences between the two groups before and after the intervention (t=2.35-4.32, P<0.05). Conclusions To study and discuss the reasons of emergency department overcrowding, it can effectively relieve the happening of emergency department overcrowding through comprehensive intervention measures such as improve the environment, optimize the process, strengthen emergency medical personnel training and management, improve the service level hospital auxiliary support system.

In this study,the L.dumoffii TEX-KL (ATCC 33343),L.dumoffii NY23 (ATCC 33279) and L.pneumophila philadelphila-1 (ATCC 33155) strains were used to explore the adhesion,invasion and intracellular growth ability in the epithelial cells.Approximately 1× 108 bacteria were pelleted,resuspended,and diluted (1 ∶ 10) in RPMI 1640 tissue culture medium.The bacteria were then added to A549 cells (1 × 105 per well) in 24-well plates to give a multiplicity of infection (MOI) of about 100.The Gimenez staining and colony counting methods were used for the determination of the strain adhesion,invasion and intracellular growth ability.It was found that in vitro growth ability of L.pneumophila philadelphila-1,L.dumoffii TEX-KL and L.dumoffii NY23 strains had no significant difference.For in vivo assay,there was also no significant difference in adhesion ability of these strains.However,the CFU counts of L.dumoffii TEX-KL strain invaded into A549 cells was 1 000 times higher than that of the other two strains.Compared with L.pneumophila philadelphila-1 and L.dumoffii NY23 strains,L.dumoffii TEX-KL strain has higher invasion ability and,therefore,higher intracellular growth ability.

Objective ERP research has always been the hot spot of the n~ve electrophysiology research.A gTowing body of research suggests that ERP can effectively reflect the cognitive function in patients with depression,evaluate the risk of suicide.This paper reviews the ERP related components with depression and related research.On the basis of the application of ERP in the depression research discusses limitations and prospects on the later development.Methods The articles were retrievaled from databases,such as P UBMED,Foreign medical information resources retrieval platform,CNKI,Wanfang database,ect,from 2013-06 to 2015-01.Index strategy TI:TI:[depression OR affective disorder] AND [suicide OR suicidal] All field:event related potential.Reading and narrowing the scope of the articles,we should understand the experimental design,such as Sample company have suicidal ideation and (or) behavior of patients with depression.Potential induced way,ERP related component detection method,etc.The articles included 43 in English and 15 in Chinese.Read the full text and retain nearly five years about depression and related literature.Results Eventually,the article includes 34 articles and 3 of in Chinese.Conclusion Associated with suicidal ideation and (or) behavior of the existing cognitive dysfunction in patients with depression,ERP related components such as:abnormal P300 and CNV,MMN.N400 have not found abnormal.This article only from the Angle of depression and related research are reviewed in this paper the characteristics of ERP,ERP of other mental illnesses among related research,and to be perfect in the future.

Objective To isolate and identify the pathogenic bacteria from peripheral blood of a patient with septicemia of unknown etiology and to analyze their drug resistance genes.Methods Two peripheral blood samples were collected from the patient after having fever.Several assays including culturing bacteria on blood agar plates by using streaking technique,Gram-staining of bacterial colonies and microscopic observation,VITEK 2-compact automatic bacterium detection and analysis system as well as a sequencing analysis of the 16s rRNA gene were performed to identify the bacterial pathogens in blood samples.Microdilution test was performed to detect the drug susceptibilities of isolated bacteria to antibiotics.Confirmatory tests were performed to detect the production of β-lactamase and extended spectrum β-lactamase by the isolated strains and the phenotypes of AmpC enzyme and carbapenemase.PCR was used to identify the β-lactamase-encoding genes in the isolated strains by using the primers of 19 common β-lactamase-,AmpC enzyme-and carbapenemase-encoding genes in Enterobacteriaceae strains and the primers of 21 annotated gene sequences encoding the β-lactamase of a Ralstonia mannitolilytica strain.The PCR products were sequenced and analyzed after T-A cloning.Results Ralstonia mannitolilytica strains were isolated from the two peripheral blood samples.The isolated strains were sensitive to ceftriaxone,cefepime,ciprofloxacin,ofloxacin,tigecycline and compound sulfamethoxazole (SMZ-TMP),but resistant to the other 11 tested antibiotics.Results of PCR amplification by using the primers of common genes encoding β-lactamase of Enterobacteriaceae strains were all negative.Fragments of genes encoding the β-lactamase of the isolated Ralstonia mannitolilytica strain were successfully amplified,which were TK49_09850,TK49_12955,TK49_14470,TK49_14495and TK49_18990.The sequences of the amplified gene fragments were not similar to those of the common β-lactamase-encoding genes in Enterobacteriaceae strains.Conclusion The patient was infected with Ralstonia mannitolilytica.The isolated Ralstonia mannitolilytica strain showed a high-level drug resistance with a noticeable diversity against different β-lactam antibiotics.The genes encoding β-lactamase of the isolated Ralstonia mannitolilytica strain were completely different to those of Enterobacteriaceae strains.

Objective: To optimize the extraction technology of Panax notoginseng and Scrophulariae radix from Rupixiao granule.Methods: With the dry extract rate and transfer rates of ginsenoside Rg1, ginsenoside Rb1 and harpagoside as the comprehensive index, the orthogonal design was adopted to investigate the effects of the amount and concentration of ethanol, extracting duration and times on the extraction technology.The contents of ginsenoside Rg1, ginsenoside Rb1 and harpagoside were determined by HPLC.Results: The optimal extraction technology was extracted twice with 8-fold amount of 60% ethanol with 2 h per time.The transfer rate of ginsenoside Rg1, ginsenoside Rb1 and harpagoside was (79.4%±1.56%), (42.62%±0.68%) and (44.89%±0.58%)(n=3), respectively.The dry extract rate was (20.99%±0.411%).Conclusion: The optimized extraction technology is stable and feasible, which can be used for extracting Panax notoginseng and Scrophulariae radix from Rupixiao granule.

Objective To analyze the effect of autologous bone marrow stem cells transplantation in treatment of patients with decompensated cirrhosis. Methodls Seventy-eight patients (aged from 26 to 67) with decompensated cirrhosis, including 56 with hepatitis B, 21 with alcoholic cirrhosis and 1 with schistosomial cirrhosis, were included. Bone marrow was aspirated from poster superior spine. After isolation and purification, the stem cells were transplanted into liver via hepatic artery. The liver function, laboratory parameters and Child-Tureotte-Pugh scores were evaluated in 2,4 and 8 weeks after transplantation. Results At the 4th week after transplantation, the level of albumin was increased obviously from (32.9±5.58) g/L to (38.32±6.45) g/L,whereas the alanine aminotransferase was decreased from (96.92±83.91) U/L to (73.48±18.46)U/L. It was revealed that the prothrombin time was decreased from (16.66±3.91) s to (15.52±3.35) s and fibronegen increased from (2. 22 ± 0. 88) g/L to (2. 58±0. 88) g/L. After transplantation, appetite was improved in 72 cases (92.3%), ascites was decreased in 70 cases (89.7%) and abdomen distention was ameliorated in 68 cases (87.2%). There was no complications related to the transplantation. Conclusion Transplantation of autologous bone marrow stem cells is a safe and effective method in treatment of patients with decompensated cirrhosis.

Objective To explore the mechanism of protecting cells from hypoxia/ reoxygenation (H/R) injury by constructing specific small interference RNA (siRNA) to inhibit NALP3 expression in rat renal tubular epithelial cells (NRK-52E).Methods (1) To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 1 h,2 h,4 h,8 h,16 h and 24 h.The activity of lactae dehydrogenase (LDH) in the culture medium,cell count and cell viability,the expression of NALP3 were determined by biochemical method,trypan blue exclusion and Western blotting.(2) The siRNA was transfected into NRK-52E.The irrespective siRNA transfected group wasused as control.NALP3 expression was examined by Western blotting.(3) The cells were divided into 4 groups:control group,H/R group,irrespective siRNA transfected group and NALP3-siRNA transfected group.To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 4 h.And the expression of NALP3 was determined by Western blotting.(4)Cellular apoptosis was examined by Annexin V/PI staining and flow cytometry.NF-κB DNA binding activity,IκB-α,Bcl-2 and Bax expression were examined by EMSA and Western blotting.Results (1)Compared with the control group,the activity of LDH significantly increased,cell count and cell viability significantly decreased (all P＜0.05).The expression of NALP3 significantly increased and peaked at 4 h after H/R.(2)The specific siRNA could efficiently inhibit NALP3 expression in NRK-52E.Compared with the irrespective siRNA transfected group,the protein expression of NALP3 was significantly down-regulated in NALP3 siRNA transfected group (P＜0.05).(3)After hypoxia 1 h and reoxygenation 4 h,the activity of LDH and the expression of NALP3 increased.Compared with the irrespective siRNA transfected group,LDH concentration in media and the expression of NALP3 significantly decreased in NALP3-siRNA transfected group.(4)After hypoxia 1 h and reoxygenation 4 h,NF-κB DNA binding activity was increased,IκB-α phosphorylation and degradation,Bcl-2 and Bax were significantly up-regulated.However,compared with the irrespective siRNA transfected group,NF -κB DNA binding activity,IκB-α degradation and Bax/Bcl-2 were significantly decreased (P＜0.05) in NALP3-siRNA transfected group.At the same time,the ratio of apoptosis was significantly increased in three groups than that in control.Compared with the irrespective siRNA transfected group,the ratio of apoptosis in NALP3-siRNA transfected group was significantly decreased (P＜0.05).Conclusions H/R induces the expression of NALP3 in NRK-52E.The synthesized siRNA can inhibit the expression of NALP3 and protect NRK-52E from hypoxia/reoxygenation injury.The mechanism may be via inhibiting the activation of NF-κB,modulating expression of Bcl-2 and Bax,as well as decreasing cell apoptosis.

Objective To investigate the induction of specific anti-tumor immune response by transfected dendritic cells (DCs) with MUC1 mRNA of human pancreatic cancer,and to provide the experimental evidences for the treatment of human pancreatic cancer with DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs),and then were identified by cell morphology and surface markers.After being transcripted and amplified,MUC1 mRNA was transfected into DCs by electroporation.The expression of MUC1 in DCs at different time points was detected by quantitative real-time PCR and Western blot.The survival rate of DCs before and after tramrfection was determined by MTT method.The induction of specific cytotoxic T lymphocyte (CTL) response by MUC1 mRNA transfected DCs was measured by 51Cr standard cytotoxicity test.The released amount of IFN-γ was evaluated by ELISA method.Results The cultured cells appeared typical characteristics with regard to morphology and phenotype (CD40 +,HLA-DR+,CD83 +,CD86 + ).After MUC1 mRNA transfection for 48 h,the expression of MUC1 mRNA of DCs reached the highest point ( 38.43 ) and the MUC1 protein expression also reached the highest point at 72 h.The survival rate of DCs was stabilized around 80％ after transfection.The DCs transfected with MUC1 mRNA could effectively induce HLA-A2+/MUC1 + specific CTL immune responses.Stimulated by pancreatic cancer cell line Capan-2 cells or the DCs transfected with MUC1 mRNA,the IFN-γ released in 24 h by MUC1 specific CTL were ( 28.44 ± 4.96 ) U/m1 and ( 16.31 ± 2.54) U/ml,respectively.The difference between the two groups was statistically significant (P ＜0.05 ).Conclusions DCs transfected with human pancreatic cancer MUC1 mRNA could induce CTLs and produce specific anti-tumor immunity.

Objective To investigate the ability of induction of specific cytotoxic T lymphocytes (CTL) stimulated by dendritic cells (DCs) co-transfected with MUC1 and survivin mRNA of human pancreatic cancer,and to provide the experimental basis for the treatment of human pancreatic cancer with multi-epitope DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) of 6 patients with pancreatic cancer.Human pancreatic cancer cell line MiaPaCa-2 was routinely cultured,after being transcripted and amplified by RT-PCR,MUC1 and survivin mRNA were co-transfected or individually transfected into DCs by electroporation,and they were named as DC-MUC1,DC-survivin,DC-MUC1 + survivin.The expression of MUC1 and survivin mRNA in DCs were detected by real-time PCR.The survival rate of transfected DCs were determined by MTT method.The lymphocyte proliferation ability was evaluated by mixed cell culture method.The Th1 cytokine releasing of antigen-specific CTLs were measured by ELISA assay.Results Mature DCs were obtained,the positive expression rates of surface markers CD40,HLA-DR,CD83 and CD86 were 34.31％,50.21％,89.17％ and 73.62％,respectively.The expression amount of MUC1 mRNA of DC-MUC1 was 36.24 ± 5.17,and the expression amount of survivin mRNA of DC-survivin was 34.53 ± 4.02,while the expression amounts of MUC1,survivin mRNA of DC-MUC1 + surviving were 31.79 ±4.26 and 14.67 ± 2.96,which were significantly lower than that in individual transfection group (P ＜ 0.05).The survival rate of DC-MUC1 + surviving was decreased in a time dependent manner,which was significantly lower than that in individual transfection group (about 50.21％ vs 80％ at 24 h,P ＜0.05).When DC/T cells ratio was 1∶ 10,1∶ 20,the autologous T cell proliferation index of MUC1 and survivin mRNA in co-transfection DC group was significantly higher than that in individual transfection group (P ＜ 0.05) ;when DC/T cells ratio was 1∶ 40,1∶ 80,the difference of proliferation index was not statistically significant.When DC/T cells ratio was 1∶ 10,after 14 d culture,the expressions of IL-2 in DC-MUC1,DC-survivin,DC-MUC1 + surviving were (892.73 ± 32.9),(713.62 ± 56.37),(1884.37 ± 95.21) pg/ml,and the expressions of granzyme B were (501.62 ± 12.30),(203.84 ± 12.55),(1193.15 ± 86.04) pg/ml ; and the expressions of IFN-γ were (981.50 ± 47.82),(696.05 ± 41.66),(2237.94 ± 189.55) pg/mL.The corresponding values in DC-MUC1 + surviving group were significantly higher than those in individual transfection group (P ＜ 0.05) ; while the difference of IL-10 was not statistically significant.Conclusions DCs co-transfected with MUC1 and survivin mRNA have a stronger ability to stimulate specific CTL in vitro than individual antigen loaded DCs.

Objective To compare the mesothelin expressions in 3 human pancreatic cancer cell lines between in vitro and in vivo and the developing speed among the subcutaneous tumors implanted with the 3 human pancreatic cancer cell lines in nude mice.Methods The human pancreatic cancer cell lines ( SW1990,BxPC3 and PANC1 ) were cultured and then were implanted subcutaneously into left axillas of nude mice.The volumes of these subcutaneous tumors were recorded every week to estimate their developing speed.The mice implanted with SW1990 and BxPC3 cells were observed for three weeks,while the mice implanted with PANC1 cell were observed for five weeks.The Western blot method was used to measure the expressions of mesothelin in the 3 kinds of cells and subcutaneous tumors,while immunohistochemical staining was applied to determine the expressions of mesothelin in 3 kinds of subcutaneous tumors.Results The sequence of quantities of expressions of mesothelin in these cell lines in vitro were BxPC3 ＞ PANC1 ＞ SW1990,and the sequence of quantities of expressions in vivo were SW1990 ＞ BxPC3 ＞ PANC1.One handrued percent of the tumors grew out successfully,and the sequence of speeds of their growth was SW1990 ＞ BxPC3 ＞ PANC1.Conclusions The mesothelin expressions among 3 kinds of pancreatic cancer cell line are different.The developing speeds of tumors originated from different subcutaneous tumors in nude mice are also different,and there is no association between them.