Objective:To Clone the silencer sequences of human stimulator of interferon genes (STING) and evaluate its activity in HEK293T and HeLa, and to preliminarily investigate the transcriptional regulatory mechanisms, screening and verifying the possible binding elements for the silencer sequences of STING.Methods:The human STING-5-1a(-124~+ 267, transcription start site, TSS: 0) and STING-5-2a(-124~+ 168) regions were amplified by PCR, subcloned into pGL3-Basic plasmid, and the luciferase activity was detected in HEK293T and HeLa; the human STING silencer region STING-5-1b(+ 169~+ 267) was subcloned into the pGL3-Control plasmid (pGL3-C-5-1b-positive/negative) and STING-5-1b is divided into two complementary elements, subcloned into pGL3-Control vector, named pGL3-C-STING 5-1b-α(+ 169~+ 209) and pGL3-C-STING-5-1b-β(+ 210~+ 267), detecting the relative luciferase activity of the above plasmid in HEK293T and HeLa. Using bioinformatics methods to predict the transcription factor binding site of human STING silencer, make site-directed mutagenesis at the predicted binding site, and detect luciferase activity. Knockdown of THAP1 and TEAD1 by a siRNA strategy and detect the expression level of STING by western blot analysis. Chromosome immunoprecipitation assay further verified the binding of transcription factor to hSTING silencer.Results:It was verified that plasmids mentioned above were constructed correctly by nucleotide sequencing. The relative luciferase increased after truncating the STING-5-1b(+ 169~+ 267) fragment. The relative luciferase activity of pGL3-C-5-1b-positive recombinant plasmid decreased ( P<0.05), compared with pGL3-Control plasmid. Among them, STING-5-1b-β(+ 210~+ 267) fragment play a major inhibitory role. Using bioinformatics software to predict that the transcription factors PR domain zinc finger protein 4 (PRDM4), Thanatos-associated protein 1 (THAP1), TEA Domain Transcription Factor 1 (TEAD1), Nuclear receptor subfamily 4 group A member 1 (NR4A1), Krueppel-like factor 4 (KLF4) and Forkhead box protein O3(FOXO3) may bind to the human STING silencer region (+ 210~+ 267). After transfecting the mutant recombinant plasmid of the transcription factors into HEK293T and HeLa, the relative luciferase activity of THAP1-Mut and TEAD1-Mut were significantly increased, suggesting that STING silencers may contain binding sites of THAP1 and TEAD1. Knockdown of THAP1 and TEAD1 by a siRNA strategy significantly enhanced the transcription activity. Chromosome immunoprecipitation assay showed that the transcription factors TEAD1 and THAP1 combined with hSTING silencer region in the cells. Conclusions:The hSTING silencer luciferase reporter plasmid was successfully constructed. By the activity comparison, it is speculated that the core silencer region of human STING is located in the + 210~+ 267 element, which may contain several potential transcription factor binding sequences.The potential binding sites for transcription factors that may be contained in the DNA, and use Western blot and chromosome immunoprecipitation assays to further confirm the combination of transcription factors TEAD1 and THAP1 with hSTING silencer, laying the foundation for subsequent research.

Objective To investigate the optimal extraction technique and antioxidant activity of curcuminoids from Curcuma longa L. Methods The optimized extraction process of curcuminoids from Curcuma longa L. with ethanol as extracting solvent were studied based on orthogonal test.The effect of each factor such as the ethanol concentration, the amount of solvent, extraction temperature and extraction time were investigated based on a single-factor experiment.The effects of extraction yield of curcuminoids under different processing conditions were compared by HPLC method. Content of curcuminoids and antioxidant activity of the extract were used as comprehensive survey index. Free radical scavenging DPPH method was used to assess the antioxidant effect. And the extraction technique parameters were optimized. Results The condition of optimal extraction technique was: ethanol concentration 60%, amount of solvent 20 folds, extraction time 2 h, extraction temperature 90 ℃. Under these conditions, the extraction yield of curcuminoids was 3. 65%. DPPH free radical scavenging yield of the curcumin extract was 79.79%. Conclusion This optimized technology is simple and have a good reappearing rate, and the extracts of Curcuma longa L.show good antioxidant activity.

OBJECTIVE To investigate nosocomial infection and antibiotic resistance of Pseudomonas aeruginosa among geriatrics patients and to instruct clinical monitoring and control infection.METHODS P.aeruginosa isolates from geriatrics patients were identified and the resistance to antibiotics was determined by K-B method.RESULTS The risk factors of nosocomial infection were underlying diseases,low immunological function,long time hospitalization,invasive treatment,and prolonged antibiotic therapy.The resistant rates to trimethoprim/sulfamethoxazole and cefotaxime in PA were 74.8% and 60.2%,respectively.The resistance rate to imipenem was 3.9%.CONCLUSIONS It is suggested to carry out proper measurement to control the spread of multiresistant strains in order to improve the treatment of PAE infection in geriatrics patients.

OBJECTIVE To investigate the resistance phenotype and clinical feature of super extended spectrum ?-lactamases(SSBLs) producing Escherichia coli in order to provide reference for the clinical application of drugs.METHODS Totally 945 strains of ESBLs producing E.coli collected from Jan 2003 to Jun 2007 were identified by API microbiological assay system.Susceptbility tests were performed by K-B methods.Improved three-dimensional tests were adopted to test ESBLs and AmpC lactamase.Test data were analyzed statistically by WHONET 5.3 software.RESULTS From them eighteren strains of SSBLs producers were detected.Between the positive for ESBLs strains and the negative strains,there were some significant differences in the antimicrobial resistance(P

Objective To study the role of D‐lactate gradient across the lung in the rapid diagnosis of pneumonia and evalua‐tion of therapeutic efficacy .Methods Patients were divided into pneumonia group (n=46) and non‐pneumonia group (n=28) in ICU .D‐lactate gradient across the lung were calculated by the difference between arterial and mixed‐venous D‐lactate concentrations before the treatment ,after 3 and 7 days of treatment .Serum procalcitonin (PCT) ,Oxygenation index ,the lung injury score (LIS) and clinical pulmonary infection score(CPIS) were recorded at the same time .Results The mean D‐lactate gradient across the lung in pneumonia group was significantly higher than that in non‐pneumonia group[(163 .84 ± 10 .72) ng/mL vs .(30 .33 ± 7 .25) ng/mL ,P<0 .01) ]before treatment .Using a cut‐off value of 106 .11 ng/mL ,D‐lactate gradient across the lung′s sensitivity for di‐agnosis pneumonia was 90 .7% and its′specificity was 75 .5% .D‐lactate gradient across the lung correlated with LIS (r= 0 .554 , P<0 .01) and CPIS(r=0 .543 ,P<0 .01) .Conclusion D‐lactate gradient across the lung correlates with lung injury and pulmonary infection positively and may be a potential biomarker for rapid diagnosis of pneumonia .

Objective To investigate the inhibiting effects of shRNA(short hairpin RNA,shRNA) on COX-2 gene expression,the cell cycle and growth of gastric carcinoma SGC-7901 cells.Methods Specific shRNA plasmid to COX-2 were constructed,and then transfected into SGC-7901 cells by lipofectamine methods.Tests were divided into three groups: untransfected gastric carcinoma SGC-7901 cells group,negative control HK group and pshRNA-COX-2 group.Gastric carcinoma SGC-7901 cells were transfected with LipofectamineTM 2000.Expression of COX-2mRNA and protein were detected with reverse transcriptional polymerase chain reaction(RT-PCR) and Western blot respectively.Cell cycle analysis and cell growth chart were detected with flow cytometry and cell count respectively.Results Compared with negative control HK group,recombinant expression vector pshRNA-COX-2 resulted in the reduction of COX-2mRNA and protein expresion by 70.1% and 43.2% respectively;cells in G0-G1 phase increased from 61.5%to 70.2%,cells in S phase decreased from 27.3% to 21.7%,and the growth of SGC-7901 cells cells was slowed significantly.Conclusions Recombinant expression vector pshRNA-COX-2 can significantly inhibit the expression of COX-2 gene,result in the increase of cells in G0-G1 phase and decrease of cells in S phase,and suppress proliferation of gastric carcinoma SGC-7901 cells.

Objective To investigate the related index of early diagnosis of acute-on-chronic liver failure caused by chronic hepatitis B.Methods 13 cases of the acute-on-chronic liver failure caused by chronic hepatitis B were collected in our department,39 cases of chronic hepatitis B were randomly paired to analyze the difference index.ROC curve analysis was conducted for the different data,then to choose the data whose AUC (area under the curve)is >0.8 to accumulate points,while the cutoff value in the light of Youden index was indentified.Each accu-mulating points data would score 1 point if it is≥cutoff value,otherwise,it would score 0 points.ROC curve analysis was conducted for the total accumulating points,the cutoff value of the total accumulating points according to Youden index was indentified,the sensitivity and specificity were calculated.Results The data of which AUC are >0.8 include glutamic-pyruvic transaminase (ALT)/Upper Limit Of Normal (ULN),neutrophil count (N),alkaline phosphatase (AKP)/ULN,glutamic-oxalacetic transaminease(AST)/ULN,direct bilirubin/total bilirubin(Dbil/Tbil),total bile acid(TBA),whose AUC were respectively 0.869,0.874,0.897,0.917,0.919 and 0.978,while their cutoff value were respectively 18.57,3.1 ×109/L,0.99,22.21,44.41%and 140.20μmol/L.The cutoff value of total accumula-ting points of the six data was 4 points,AUC was 0.987,Youden index was 0.949,and their sensitivity for ealier diag-nosis for the acute -on -chronic liver failure caused by chronic hepatitis B was 100%,specificity was 94.87%. Conclusion The accumulating points of the six data aboved (ALT/ULN,N,AKP/ULN,AST/ULN,Dbil/Tbil,TBA) will help the early diagnosis of acute-on-chronic liver failure caused by chronic hepatitis B.

Objective To analyze the effects of Ileus tube (IT) along with somatostatin (SS) used for treatment of patients with severe acute pancreatitis (SAP).Methods Under conventional treatment,75 patients with SAP were divided into three groups as per different additional treatments,namely group A (IT and SS),group B (nasogastric decompression tube and SS),and group C (IT alone),and the therapeutic efficacies of those treatments were evaluated in respects of improvement of physical signs and symptems,dynamic changes in decompression drainage and prognosis.Results Therapeutic efficacy was 100％ in group A,84％ in group B and 80％ in group C.The difference between group A and B was x2 =8.26 (P ＜0.01) ; group B vs.C was x2 =0.38 (P ＞ 0.05).The physical signs and symptoms in group A were improved more rapid than those in group B and C (P ＜ 0.05),but there was no significant differences in those signs and symptoms between group B and group C (P ＞ 0.05).In comparison of decompression drainage,the t value of group A vs.group B was 2.14,group B vs.group C was 3.83,and group A vs.group C was 2.23 (P ＜ 0.05).As cure rate of patients with SAP in hospital on the 14th day,rate of transferring to surgical treatment as a last resort and mortality in group A were compared with group B and C,the differences were statistically significant (P ＜ 0.05),while group B vs.C,the difference was not statistically significant (P ＞ 0.05).Conclusions Application of IT combined with SS can significantly improve the condition of patients with SAP,thereby reducing the operation rate,shortening hospital stay,lowering mortality and improving the outcome,and it is worthy of clinical popularization.

Objective To investigate the clinical features and CT appearances of primary adenosquamous carcinoma (ASC)of the liver and to improve the understanding of this disease for the sake of misdiagnosis.Methods The clinical features and CT appearances in 7 patients proved by operation and pathology were reviewed,retrospectively.Results All of the patients had dull pain in upper abdo-men,fever,chills and a long history of cholangitis and biliary calculi.In addition,all patients had not the history of hepatitis B and liver cirrhosis.And the serum AFP level was normal.The single lesion was found in every patient including 4 lesions in the left lobe of liver and the other 3 in the right lobe.Plain CT showed all masses with hypodensity,heterogeneity and unclear edge in liver,and multiple irregular and more hypodense areas in lesions were found.Slight heterogeneous honeycomb-like enhancement in the arterial phase was showed.In the venous phase,persistent honeycomb-like enhancement with uneven separations,nodular bulge and hypo-dense necrotic area was found.In the delayed phase,further honeycomb-like enhancement with hypodense necrotic area and obvious-ly enhanced nodular bulge was showed.The bile ducts in the liver and around the mass were dilated and had pneumatosis in company with lithiasis in choledochus and intrahepatic bile duct in 5 patients.Conclusion Primary hepatic ASC has certain clinical character-istics in older patients.The CT characteristic features included:honeycomb-like lesions with persistent,heterogeneous,delayed en-hancement and heterogeneous separation,uneven inner edge and enhanced nodular bulge.

OBJECTIVE To explore the drug-resistant situation of clinical isolated Escherichia coli in our hospital from Jun to Dec in 2007,the aim is to offer evidence for drug-resistant monitoring and clinical antibiotics usage in our hospital. METHODS All clinical specimens isolated and cultured from patients were identified by using the automatic microorganism analyzer VITEK-2 as well as bacteria's drug susceptibility tests were performed using counterparts panel. RESULTS A total of 352 strains E. coli were isolated. The isolated ratio of extended spectram-?-lactamases (ESBLs) producing E. coli was 66.2% (233/352),no ESBLs producing E. coli was 33.8% (119/352). From them 111 strains E. coli were isolated in sputum and 89 strains were ESBLs producing and 22 strains were no ESBLs producing; 111 strains E. coli were isolated in urine and 62 strains were ESBLs producing and 49 strains were no ESBLs producing. The drug-resistance difference was obvious between them as well as between strains isolated from different sites. Better to select piperacillin/tazobactam,cefotetan,ertapenem,imipenem,amikacin,and nitrofurantoin to cure all isolated E. coli infection.CONCLUSIONS The drug-resistance of ESBLs producing E. coli is severe (66.2%) so that hospital administers should strengthen antibiotics usage management and improve antibiotics rational usage,inorder to decrease occurrance of bacterial resistance.