Objective To investigate the technique and clinic value of non-operation management for ERCP-related peripancreatic and retroperitoneal abscess. Methods Five patients with post-ERCP peripancreatic and retroperitoneal abscess were reviewed. Guided by ultrasonic or CT, all the 5 pa-tients underwent puncture and the drainage tube was disposed to the lowest place of abscess. Non-op-eration managements for patients also included the use of anti-inflammatory drugs and enzyme activity inhibition drugs. Results All patients responded to the draining treatment and discharged from hospi-tal after complete recovery. There was no conversion to surgical intervention. Mean draining duration was 52. 4(20-90)d and average hospital stay was 91.8(35-165)d. Conclusion Puncture and drainage management is an effective and safe approach for post-ERCP peripancreatic and retroperitoneal ab-scess. It has advantages of less trauma, less pain, fast recovery and low rate of complications. Punc-ture point should be situated at the bottom or lowest position of abscess and drainage can achieve the best results.

Objective To investigate the role of D-dimer in the diagnosis and evaluation of mycoplasma pneumonia.Methods The positive rate and concentration of D-dimer in mycoplasma pneumonia,bacterial pneumonia,viral pneumonia and healthy controls were detected and compared.The positive rate and concentration of D-dimer in patients with systemic inflammatory response syndrome(SIRS)and without SIRS were observed.Changes of D-dimer concentration in patients with mycoplasma pneumonia before and 2 weeks after treatment were observed.Results Compared with bacterial pneumonia group(16.7%),the viral pneumonia group(20.0%)and healthy control group(0.0%),the positive rate of D-dimer in mycoplasma pneumonia group(41.6%)was significantly higher(x2=5.625,4.158,17.308,all P<0.05).The positive rate of mycoplasma pneumonia group with SIRS(64.0%)was significantly higher than that in other groups(x2=17.308,P<0.05).The D-dimer levels of the mycoplasma pneumonia group [(324.4±125.3)μg/L],bacterial pneumonia group[(282.3±95.4)μg/L] and viral pneumonia group[(293.1±92.3)μg/L]were significantly higher than that in the healthy controls[(72.9±22.4)μg/L](t=10.878,11.704,12.698,all P<0.05).The concentration of D-dimer[(381.4±129.4)μg/L] in the mycoplasma pneumonia group with SIRS was significantly higher than that in the other groups(t=2.668,P<0.05).Compared with before treatment,the concentration of D-dimer after treatment in the two groups was significantly lower[(129.3±65.3)μg/L,(89.7±28.6)μg/L](t=2.582,P<0.05).Conclusion The level of D-dimer in children with mycoplasma pneumonia was significantly increased,and the severity of the disease could be reflected.

Objective To assess the volume of epicardial adipose tissue(EAT) and intrathoracic adipose tissue(IAT) and the correlation of EAT,IAT and the EAT/IAT ratio with the severity of coronary artery atherosclerosis in patients with metabolic syndrome(MS) and coronary heart disease(CHD).Methods Ninty-seven MS subjects without coronary atherosclerosis and one hundred and eighteen MS subjects with CHD were enrolled in this study.The volumes of EAT,PAT and IAT were measured using axial data from base to apex traced manually with a dedicated semiautomatic software program-volume.Results (1)Compared with MS subjects without coronary atherosclerosis,MS subjects with CHD had significantly increased age[(60.6± 1 1.4)years vs.(57.9 ± 8.7) years,P=0.001],positive family history of cardiovascular disease(29.7％ vs.21.6％,P=0.03),EAT[(98.3±41.4) cm3 vs.(82.2±39.7) cm3,P=0.001],IAT [(171.3±64.1) cm3 vs.(156.2±48.1) cm3,P=0.001] and HbA1c[(7.1 ± 1.8)％ vs.(6.6±2.3)％,P=0.02],but siginificantly reduced total cholesterol[TC,(4.9 ± 1.2)mmol/L vs.(5.4 ± 1.0) mmol/L,P=0.003],low-density lipoprotein cholesterol[LDL-C,(3.0±1.1) mmol/L vs.(3.6±1.0) mmol/L,P=0.03] and high-density lipoprotein cholesterol[HDL-C,(1.0 ± 0.4) mmol/L vs.(1.1 ± 0.3) mmol/L,P=0.04].(2) In MS subjects without coronary atherosclerosis,the volumes of EAT,pericardial adipose tissue(PAT) and IAT were associated with gender,body mass index(BMI),waist circumference(WC),diabetes mellitus and hyperlipidemia(all P＜0.05); however,in MS subjects with CHD,the volumes of EAT,PAT and IAT were associated with BMI and WC (both P＜0.05).(3) The volumes of EAT and IAT were correlated with calcification grades(r values were 0.45 and 0.50,P values were 0.017 and 0.013,respectively) and Gensini score(r values were 0.476 and 0.563,P values were 0.015 and 0.017,respectively) instead of coronary artery calcifacation score(CACS).Moreover,the EAT/IAT ratio was negatively correlated with CACS(r=-0.321,P=0.028).Conclusions Compared with MS subjects without coronary atherosclerosis,MS subjects with CHD have significantly increased EAT and IAT volumes.In our subjects,the volumes of EAT,PAT and IAT were associated with BMI and WC.Only in MS subjects with CHD,the volumes of EAT and IAT were correlated with caleification and Gensini score.

Objective To observe the ultrastructure and absorption function of colon mucosa in rat with ultra-short bowel syndrome. Methods Totally 30 SD rats were randomly divided into three groups: ultra-short bowel group (90%-95% of the intestine was surgically resected, n = 10), sham group (n = 10), and normal control group (n = 10). All animals were given with enteral nutrition. Scanning electron microscopy was performed 21 days later to observe the morphology of mucosal surface, and transmission electron microscopy was performed to observe the ultrastructural changes of intestinal epithelial cells. The absorption of colon to water, carbohydrates, and amino acid determined after 3 hours of closed perfusion of the colon with D-xylose solution and 15N-glycine on the continuous cycle of colon. Results As shown by the transmission electron microscopy, compared with the normal control group, rats in the ultra-short bowel group showed significantly decreased goblet cells on colonic mucosl surface, increased epithelial cells, longer and denser microvillus, increased area of membrane surface, increased number of cell-cell junctions, increased number of desmosome, tight junction, and gap junctions, higher development of endoplasmic reticulum and dictyosome, and increased number of mitochondria. As shown in the screening electron microscopy, compared with the normal rats, rats in the ultra-short bowl group had significantly deeper colon folds, thicker mucous membrane, increased number of bay openings, and longer and denser microvillus-like structures inside bays. The capability of water absorption was signicatnly higher in the ultra-short bowl group than in the sham group and normal control group (P = 0. 000) . The absorption rates of xylose and 15 N-glycine were also significantly higher in the ultra-short bowl group than in the control group (P ＜ 0. 01). Conclusions The absorption capability can be compensatively increased in rats with ultra-short bowel syndrome. Decreased apoptosis of colon mucosa cells, increased absorption cells, hyperplasia of microvilli, increased area of the membrane surface,and increased number of mitochondria may constitute its material and energy bases.

Objective This study is to analyse the clinical feature and risk factors of morbidity after pulmonary resection for lung cancer in patients older than 70 years. Methods The clinical records of 222 patients older than 70 years who had undergone pulmonary resection for their lung cancer was reviewed. The patients were divided into 3 groups: group Ⅰ including the patients who had severe postoperative complications, group Ⅱ including the patients who had mild complications and group Ⅲ including the patients who had no complications. Moreover, the definitions were made that group A1 = group Ⅰ+ Ⅱ , group B1 = group Ⅲ, group A2 = group Ⅰ and group B2 = group Ⅱ + Ⅲ. Univariate analyses and multivariate binary logistic regressions relating postoperative morbidity to risk factors were performed between the group Al and Bl, A2 and B2, resulting in the identification of the independent risk factors for overall morbidity and major morbidity. Results Preoperative comorbidity was recorded in 161 patients (72.5%). Lobectomy (64.9% ) was the predominant surgical procedure. The median number of dissected LN was 14, with the range of 0 to 57. The overall morbidity was 63.5% , including major morbidity of 13.5%. Perioperative mortality was 1.8% (4 cases). The results of binary logistic regression analyses indicated that the independent risk factors for overall morbidity were preoperative weight loss (P =0.020), ASA score (P＜0.001), MVV (% predicted) (P=0. 020 ) and the number of dissected LN ( P = 0.004 ). The independent risk factors for major morbidity were ASA score ( P =0.003), MVV (% predicted) (P= 0.018) and the location of tumor (P=0.007). Conclusion Preoperative weight loss and numbers of dissected mediastinal lymph nodes were risk factor for lung cancer patients older than 70 years, Proper perioperative management for the elderly patients with high ASA score, low MVV (% predicted) or central tumor, could reduce the major postoperative morbidity.

Objective: To investigate the multi-imaging diagnostic values, especially MSCT technology in patients with congenital aortic diverticulum with its clinical application. Methods: The MSCT ifndings in 12 patients with congenital aortic diverticulum were retrospectively analyzed. Results: There were 9 patients with right aortic arch and 1 with left aortic arch, all of them having coexisted aberrant subclavian artery which initially dilated like aneurysm by diverticulum changing (Kommerell diverticulum), and there was 1 patient with incomplete double aortic arch with atresia of left arch combining retro-esophageal aortic diverticulum (RAD) and 1 patient with ducts diverticulum. Echocardiogram only made the suggestive diagnosis of speeding up blood lfow or right aortic arch in 4 patients. While MSCT accurately displayed the diverticulum for the location, morphology and with or without other complications. The post-eroanterior chest radiograph indicated “double aortic node” as the special sign in 8 patients. The echocardiogram, X-ray and MSCT for correctly diagnosing the aortic diverticulum were as 0, 72.7% and 100% respectively. Conclusion: MSCT is a rather ideal non-invasive diagnosing method for aortic diverticulum, meanwhile X-ray could also make suggestive diagnosis; if MSCT and X-ray joint with echocardiogram examination may provide the effective supplement for valve structure and hemodynamics condition in relevant patients.

Objective To evaluate the possibilities of using circulating miR-155-5p, miR-21-5p, miR-29a and miR-142-5p as biomarkers for the diagnosis of active pulmonary tuberculosis (TB).Methods Plasma samples were collected from 60 healthy subjects, 40 patients with active pulmonary TB and 20 sub-jects with latent tuberculosis infection ( LTBI) to extract miRNAs.The levels of miR-155-5p, miR-21-5p, miR-29a and miR-142-5p in plasma samples were detected by using reverse transcription-quantitative PCR. Results The levels of miR-155-5p, miR-21-5p and miR-29a in patients with active pulmonary TB were re-spectively 10.13, 7.34 and 2.74 times as much as those in healthy subjects(P<0.05).No significant differences with the level of miR-142-5p were observed between the two groups.The receiver operating char-acteristic (ROC) curve analysis of miR-155-5p, miR-21-5p and miR-29a in patients with active pulmonary TB and healthy subjects showed that the areas under the curve (AUC) were respectively 0.905, 0.830 and 0.687.The level of miR-155-5p in patients with LTBI was 3.1 times than that in healthy subjects ( P<0. 05).No differences with miR-21-5p, miR-29a and miR-142-5p were found between patients with LTBI and healthy subjects.The levels of miR-155-5p, miR-21-5p and miR-29a in patients with active pulmonary TB were respectively 3.26, 6.69 and 1.98 times than those in patients with LTBI (P<0.05).The rate of LTBI was 40.58%in people who were in close contact with patients with active pulmonary TB.Conclusion Sig-nificant differences with the levels of miR-155-5p, miR-21-5p and miR-29a were observed among healthy subjects, patients with active pulmonary TB and patients with LTBI, but no difference with the level of miR-142-5p was observed among them.miR-155-5p, miR-21-5p and miR-29a could be used as the potential bio-markers for the diagnosis of active pulmonary tuberculosis and latent tuberculosis infection.People who were in close contact with patients with active pulmonary TB could have a higher LTBI rate.

ObjectiveTo determine whether aadA2 gene can be translated from the ATG triplet,which there was no plausible ribosome binding site preceding it,and synthetized a functional protein in class 1 integron.MethodsSite-specific mutagenesis was used to construct aadA2 gene cassette with different start codons,together with their upstreamed promoters of variable regions were cloned into plasmid pACYC184 respective.The constructed plasmids were then transfored into Escherichia coli JM109,Western blot was used to detect the translation products of aadA2 gene with different start codons.Broth microdilution method was used to detect the minimal inhibitory concentrations to streptomycin in Escherichia coli JM109 containing aadA2 gene with different start codons.ResultsaadA2 gene can initiate translation from both ATG and GTG triplets in aminoacyl -3-adenylyltransferase protein synthesis,though there was no plausible ribosome binding site preceding the ATG triplet.Besides GTG and ATG triplets,there was other start codon downstream of the GTG triplet in aadA2 gene.The translated products that initiated from the start codons that described above were all functional AAD(3) proteins that can be detected by anti- aminoacyl -3-adenylyltransferase polyclonal antisera in Western blot and conferred different resistance levels to streptomycin in E.coli.ConclusionWhen inserted as the first gene cassette in class 1 integron,aadA2 gene can initiate translation from ATG triplet and synthetized a functional protein,though there was no plausible ribosome binding site preceding it.This structural characterization of class 1 integron can initiate translation of the open reading frame harbored in gene cassette that integrated into class 1 integron,though there was no plausible RBS preceding the start codon.This make class 1 integron be more convenience to express the genes that capture from environment.

ObjectiveTo prepare antiserum specific to aminoacy1-3″-adenylyltransferase [ AAD (3″) ],and to explore the application value of the prepared antiserum in detecting the expression levels of aadA2 gene that downstream of 8 different promoters (PcS,PcH2,PcH1,PcW,PcS-P2,PcH2-P2,PcH1P2 and PcW-P2 ) of variable regions in class 1 integron.MethodsaadA2 gene was amplified by polymerase chain reaction(PCR) and cloned into the expression plasmid pET19b.After inducing,the recombined aminoacy1-3″-adenylyltransferase[ AAD(3″)] with His-tag was expressed,purified and immunized rabbits to get anti- AAD(3″) specific serum.The prepared antiserum was used to detect the translation levels of aadA2 gene that downstream of different promoters of variable regions in class 1 integron by Western blotting (WB).Broth microdilution method was used to detect the minimal inhibitory concentrations (MIC) to streptomycin in Escherichia coli JM109 with aadA2 gene downstream of different promoters of variable regions.ResultsRecombined AAD (3″) expression plasmid pET19b-aadA2 was constructed successfully and was verified by sequence analysis.After transformed into E.coli BL21 ( DE3 ),a resoluble recombined AAD(3″) high expression strain was obtained.After fermentation,recombined AAD(3″) was purified and immunized rabbits.The anti- AAD(3″) specific serum was obtained with titer ＞ 1∶100 000.WB was used to detect the expression levels of AAD (3″),the translation product of aadA2 gene,that downstream of 8 different promoters of variable regions.The relative expression level of AAD (3″) that downstream of PcW was assumed to be 1,then the relative expression levels of AAD(3″),which all were detected 3 times independently,that downstream of PcS,PcH2,PcH1,PcS-P2,PcH2-P2,PcH1-P2 and PcW-P2 were 12.9±2.3,9.1±1.0,2.0±0.4,16.0±1.3,14.1 ±1.3,10.5±0.7 and 8.9 ±1.7 respective.Very different expression levels of AAD (3″) that downstream of different promoters of variable regions were obtained( F =32.421,P ＜ 0.01 ).The mean values of MIC,which all were detected 3 times independently,to streptomycin in E.coli JM109 with aadA2 gene downstream of PcS,PcH2,PcH1,PcW,PcS-P2,PcH2P2,PcH1-P2 and PcW-P2 were 256,256,64,128,32,128,4 and 64 mg/L respective.These results indicated the different expression levels of aadA2 gene that downstream of different promoters of variable regions can confer their host bacteria different resistance levels to streptomycin.Conclusions Resoluble recombined AAD(3″) is purified successfully and high titer anti- AAD(3″) specific antiserum is obtained from the immunized rabbits.This laid foundation for further investigation on the correlationship between the expressions of intI1 gene and the gene cassettes within variable regions.The expression levels of antibiotic gene cassettes that downstream of different promoters of variable regions are very different,so are the very different antibiotic resistance levels of the host bacteria.Therefore more attentions should be paid to the researches on the classification of promoters of variable regions when molecular epidemiology studies on the class 1 integrons in clinical isolates were conducted.

Objective To develop a simple high-resolution melting ( HRM) analysis method for differentiation of Pc and P2 variants in class 1 integron.Methods DNA fragments containing Pc and P2 variants were amplified from plasmids pACW ( PcW ) and pACWP2 ( PcW-P2 ) respectively , then these purified PCR products and P 2 promoters were analyed full-length amplicon by HRM .Eight DNA fragments containing different Pc promoters were amplified and site-specific mutated from plasmids pACS ( PcS ) , pACH2 ( PcH2 ) , pACH1 ( PcH1 ) , pACW ( PcW ) , genomic DNA of Klebsiellar pneumonia HS07-68 (PcWTGN-10)and HS05-1792(PcH2TGN-10)respectively.The purified PCR products and eight Pc variants were characterized by HRM analyses of an unlabeled probe and full-length amplicon.This assay was applied to the differentiate Pc and P2 variants in 109 class 1 integrons from 95 urine clinical Escherichia coli isolates in Huashan Hospital during 2004 -2007.The differentiation results were compared with that determined by direct sequencing .Results P2 promoter with a significant higher melting temperature ( Tm ) can be identified by HRM analysis clearly .P2 promoters were identified in 2 class 1 integrons and consistent with direct sequencing results .Eight Pc variants were classified into three groups: PcS, PcSTGN-10 , PcW, PcWTGN-10, PcH1, PcH1TGN-10.Using direct HRM analysis.PcH2, PcH2TGN-10 were classified into four groups:PcS, PcH1, PcH2, PcW, PcSTGN-10 , PcH1TGN-10 , PcH2TGN-10 , PcWTGN-10 according to the melting curves of the unlabeled probe .Combined the HRM analyses of the whole amplicon and unlabeled probe , the eight Pc variants can be differentiated from each other .Five different Pc variants, PcS, PcW, PcH1, PcH2TGN-10 and PcWTGN-10 , were identified and consistent with direct sequencing results .Conclusions This developed a simple Pc and P 2 variants differentiation method via simultaneous HRM analyses of an unlabeled probe and full-length amplicon .This method is cost-effective and accurate , could be used in differentiation of Pc and P2 variants of class 1 integrons in clinical isolates .