"Laparoscopic ""end-corner"" anastomosis in double-stapling technique for mid-low rectal cancer resection" / 中华普通外科杂志

Objective To observe clinical efficacy and explore clinical value of a modified procedure of double-stapling technique for mid-low rectal cancer.Methods Clinical data of patients undergoing laparoscopic anterior resection at the Department of General Surgery,the First Affiliated Hospital of Soochow University from February 2011 to February 2015 was analyzed retrospectively.According to the different ways in doing double-stapling technique,we divided patients into modified group (51 cases) and conventional group (74 cases).Parameters were compared between the two groups as general considerations,oncologic outcomes.Data were analyzed by SPSS 17.0 software packet,using t and x2 inspection.Results The difference of the general data of two groups was not statistically significant (P ＞ 0.05).Operation time in the modified group was longer than that of the conventional group [(169 ± 23) vs.(150 ±42)min,t =-3.150,P ＜0.05],but it had shorter drainage tube indwelling days [(7.9 ±2.9)d vs.(10.8±11.6)d,t=1.999,P＜0.05] and length of hospital stay after surgery [(10.0±3.6)d vs.(13.3 ± 13.7) d,t =1.025,P ＜ 0.05].The incidence of anastomotic leakage (2.0％ vs.18.9％,x2 =4.402,P ＜ 0.05) and tenesmus(3.9％ vs.17.6％,x2 =4.110,P ＜ 0.05) in the modified group was less than that of the conventional group.The difference in those areas was not statistically significant (P ＞ 0.05),such as intraopretive blood loss,per-anal exhaust time,consumption of liquid diet time,anastomotic bleeding,intestinal obstruction,reoperation for neostomy and infections.Conclusions Compared with traditional laparoscopic anterior resection,End-Corner anastomosis has the benefits of less postoperative anastomotic leakage and fewer low anterior resection syndrome.

Modification and efficacy observation of laparoscopic dual anastomosis for mid-low rectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore a new procedure of laparoscopic dual anastomosis for mid-low rectal cancer to reduce postoperative complications.</p><p><b>METHODS</b>Clinical data of 56 patients with mid-low rectal cancer undergoing laparoscopic rectal cancer resection(modified double-stapling technique, MDST, modification group) in the Department of General Surgery, the First Affiliated Hospital of Soochow University from February 2010 to June 2014 were compared with the data of 64 patients with mid-low rectal cancer (conventional double-stapling technique, DST, convention group) in the same period based on gender, age, tumor size, the distance from lower edge to the dentate line and tumor staging, etc. Patients in the modification group received operation as follows: (1) the rectum distal end was closed vertically instead of horizontally. (2) the anastomosis was conducted in an "end-corner" approach. (3) upper corner of the closed line in the distal end of rectum was removed. (4) the lower corner of closed line in the distal end of rectum was removed using vascular occlusion clamp method. (5) two T-shaped interchanges ("dangerous triangle") of stapled sutures formed after anastomosis were strengthened with absorbable suture. Patients in the convention group received laparoscopic dual anastomosis using conventional method: two corners and "dangerous triangles" were kept without any treatment. The clinical outcomes of two groups were analyzed retrospectively.</p><p><b>RESULTS</b>The intraoperational blood loss, postoperative drainage volume, postoperative anastomotic stoma bleeding, bowel function return and hospital stay were not significantly different between the two groups (all P>0.05). As compared to the convention group, the modification group had longer operation time [(211 ± 91) min vs. (174 ± 57) min, P<0.05], lower incidence of postoperative anastomotic leakage [1.8%(1/56) vs. 12.5% (8/64), P=0.030], lower tenesmus rate [3.6% (2/56) vs. 14.1% (9/64), P<0.05], less postoperative stoma re-creation [0 vs. 9.4% (6/64), P<0.05].</p><p><b>CONCLUSION</b>Modified laparoscopic dual anastomosis for mid-low rectal cancer can significantly reduce the incidence of post-surgical complications such as anastomotic leakage.</p>

A non-controlled, multicenter open-label study to evaluate the safety, tolerability and efficacy of caspofungin in the treatment of invasive candidiasis and esophageal candidiasis / 中国感染与化疗杂志

Objective This study was designed to evaluate the safety ,tolerability and efficacy of intravenous caspofungin for treatment of invasive candidiasis and esophageal candidiasis in Chinese adults .Methods This was a non-controlled ,multicenter ,candidiasis .All the 63 patients were included in the safety set (SS) and the full analysis set (FAS) .In the SS ,19 SAEs occurred in 14 patients .All these SAEs were unrelated to caspofungin .There were 73 caspofungin-related non-serious AEs in 31 patients (49 .2% ) .Five patients (7 .9% ) had both clinical AEs and laboratory abnormalities .Eight patients (12 .7% ) had clinical AEs (mainly rashes) ,and 27 patients (42 .9% ) had laboratory abnormalities ,mainly increases in liver enzymes alanine transaminase and aspartate transaminase and reduction in blood potassium .About 91 .7% of the clinical AEs were mild to moderate .Treatment was discontinued in 1 patient (1 .6% ,1/63) due to AEs .The overall efficacy was 58 .1% (36/62) in the FAS and 70 .0% (35/70) in the per-protocol set (PPS) .In the FAS ,the therapeutic efficacy was 57 .6% (34/59) for invasive candidiasis and 66 .7% (2/3) for esophageal candidiasis .In the PPS , the therapeutic efficacy was 68 .8% (33/48 ) for invasive candidiasis and 100% (3/3 ) for esophageal candidiasis .Conclusions The AEs of caspofungin were mostly mild to moderate in the treatment of invasive candidiasis and esophageal candidiasis in Chinese adults .Only one patient terminated therapy due to drug-related AE .Caspofungin is safe and effective for the treatment of invasive candidiasis and esophageal candidiasis in Chinese adults .

Expression and clinical significance of Rho GDP dissociation inhibitor 2 in pancreatic cancer / 中华胰腺病杂志

Objective To investigate the expression and clinical significance of Rho GDP dissociation inhibitor 2 (Rho-GDI2) in pancreatic cancer.Methods Immunohistochemistry and RT-PCR were used to evaluate the expression of Rho-GDI2 in 60 pairs of pancreatic cancer tissues and adjacent pancreatic tissues,and its correlation with clinicopathological parameter of pancreatic cancer was also analyzed.Results The expressions of Rho-GDI2 mRNA were 0.661 ± 0.021 and 0.199 ± 0.023 in pancreatic cancer tissues and adjacent pancreatic tissues,and the positive rate of Rho-GDI2 protein expression were 73.3％ (44/60) and 41.7％ (25/60),the positive rate of Rho-GDI2 expression in pancreatic cancer tissues was higher than that of adjacent normal tissues,and the difference between the two groups was statistically significant (P ＜ 0.01).The expression of Rho-GDI2 was strongly correlated with tumor size,differentiation,staging,lymph node metastasis,vascular invasion (P＜0.05),but it was not associated with gender,age,tumor location (P ＞0.05).Conclusions Rho-GDI2 expression is up-regulated in pancreatic cancer,and is strongly correlated with the malignant biological behavior of pancreatic cancer.

The efflux pumps in clinical Acinetobacter baumannii isolates / 中华微生物学和免疫学杂志

Objective To study the expression of active efflux pump AdeABC,AdeIJK,AdeFGH,AbeM,AbeS,CraA,MdtL in clinical Acinetobacter baumannii isolates and whether the efflux pumps confers resistance to antibiotics.Methods Thirty-two multi-drug resistant Acinetobacter baumannii islates and 10 sensitive isolates were collected.Genes of the exporter protein were amplified by PCR.The expression of adeB,adeJ,adeG,abeM,abeS,craA,mdtL were examined by real-time fluorescence quantitative RT-PCR.The controlling genes adeRS and adeL were amplified by PCR and sequenced.Results The positivity rates of adeB,adeJ,adeG,abeM,abeS,craA,mdtL were 100％,100％,100％,96.88％,100％,100％ and 93.75％ respectively in 32 multi-drug resistant Acinetobacter baumannii isolates,and all 100％ in 10 sensitive isolates.The difference of expression of adeB,abeM and mdtL were significant( P＜0.001,P =0.001,P=0.013) between 21 multi-drug resistant isolates of C clone and 10 sensitive isolates.The mutations of adeRS existed in 2 multi-drug resistant isolates,no point mutation of adeL.Conclusion The expression of AdeABC,AbeM and MdtL may involved in the resistant mechanisms of the clinical Acinetobacter baumannii islates.

Influence of different extracellular DNA levels in Staphylococccus epidermidis isolates on the ability of biofilm formation / 中华微生物学和免疫学杂志

ObjectiveTo determine the ability of biofilm formation of Staphylococcus epidermidis isolates and study the influence of different extracellular DNA(eDNA) levels in S.epidermidis isolates on the ability of biofilm formation.MethodsDetect the biofilm-formation ability of 227 S.epidermidis isolates with adhesion assays,amplify the icaA gene fragment with PCR.The S.epidermidis isolates were divided into biofilm formation(BF) group and non-biofilm formation (NBF) group according to adhesion assays and icaA amplification.Detect eDNA levels of S.epidermidis in planktonic culture and microtitre plate static culture.The eDNA in S.epidermidis biofilms stained by AO-PI was observed by CLSM.Results26 isolates were positive in adhesion assays and 32 isolates existed icaA gene among 227 S.epidermidis isolates.Select 20 isolates with positive adhesion assays and positive icaA amplification for BF group.Select 19 isolates with negative adhesion assays and negative icaA amplification for NBF group.The eDNA levels were (32.2±10.1)μg/ml,(33.6±11.9) μg/ml,(34.3±10.0) μg/ml in BF group when cultured in planktonic condition for 2,4,6 h,while the eDNA levels in NBF group were (28.7±8.9) μg/ml,(31.5±11.7) μg/ml,(31.8±12.7) μg/ml respectively.There were no significant differences between the two groups for these three phases(P＞0.05),though the eDNA levels of BF group were higher than that of NBF group.The eDNA levels were (740.0±264.4) ng/A600 in BF group when cultured in static microtitre plate,higher than that of NBF group,(80.1 ±31.1) ng/A600,and the difference between these two groups was significant.The eDNA in BF isolate Y36 biofilms could be visualized by staining with AO and PI when observed by CLSM,while neither biofilm structure nor eDNA appeard when NBF isolate Y26 was cultured for 24 h.ConclusionS.epidermidis isolates have the ability of biofilm formation.eDNA is one of the important matrix components in the S.epidermidis biofilm-forming process.The eDNA of static culture in microtitre plate was more efficient than planktonic culture in the case of estimating the ability of biofilm formation of S.epidermidis.

Association between Aia379Val and Val279Phe gene polymorphism of platelet-activating factor acetylhydrolase susceptibility and prognosis of sepsis / 中华微生物学和免疫学杂志

ObjectiveTo determine the allele frequency and genotypic distribution of plateletactivating factor acetylhydrolase (PAF-AH) gene polymorphism in the patients with sepsis. Methods Ala379Val,Val279Phe site genotypes were determined in patients (n=66) and healthy controls(n=68) by means of restriction fragment length polymorphism analysis of polymerase chain reaction products,DNA sequencing was used to detect the PCR product containing allele gene polymorphism.SPSS13.0 statistical software was use to analyze.Results All the samples by PCR-RFLP analysis of the PAF-AH gene in the Ala379Val site have three kinds of genotypes: in 66 cases of sepsis group there were 1 homozygous Val/Val type,19 heterozygous Val/Ala type,46 homozygous Ala/Ala type.In 68 cases of control group there were 2 Val/Val type,22 Val/Ala type,44 Ala/Ala type.The Ala379Val allele frequency and genotypic distribution in the patients with sepsis was not significantly different from those in the healthy controls.No statistically significant difference was observed between the survival group and the death group ( P＞0.05 ).PAFAH gene of Va1279Phe polymorphism could have three kinds of genotypes.All 66 patients in the sepsis group were the homozygous Val/Val type.Control group 68 cases,only one case was homozygous Phe/Phe type,and the others were homozygous Val/Val type,not found heterozygous Val/Phe type.The Val279Phe genotypic distribution and allele gene frequency in the patients with sepsis was not significantly different from those in the healthy controls; no statistically significant difference was observed no statistically significant difference was observed between the survival group and the death group ( P＞0.05 ).ConclusionNo associations were found between PAF-AH gene Ala379Val and Val279Phe polymorphisms and sepsis susceptibility,prognosis and severity.

Diagnosis value of soluble triggering receptor expressed on myeloid cells-1 in critically ill patients complicated with sepsis / 中华传染病杂志

Objective To investigate the diagnostic value of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) in critically ill patients complicated with sepsis. Methods Fifty-six cases of systemic inflammatory response syndrome (SIRS) who admitted in intensive care unit (ICU) of the Second Hospital of Tianjin Medical University between May 2009 and June 2010 were recruited. The SIRS patients were divided into two groups: sepsis group (n= 32) and non-sepsis group (n= 24). The levels of procalcitonin (PCT), C-reactive protein (CRP) were measured within 24 hours of hospitalization.Levels of sTREM-1 were measured by enzyme linked immunoabsorbant assay (ELISA). Based on the receiver operating characteristic curve (ROC), cut-off value, sensitivity and specificity of sTREM-1 in diagnosis of sepsis were calculated. Comparison between groups was done by t test and Mann-Whitney U test. Count data were compared by chi square test. ResultsThe serum level of sTREM-1 were significantly higher in sepsis group than that in non-sepsis group (250.9 ng/L vs 103.6 ng/L, Z= 12. 864,P=0. 002). Areas under the curve (AUC) of sTREM-1, PCT and CRP were 0. 935, 0. 891 and 0. 602, respectively. With a cut-off value of 135.0 ng/L, the sensitivity and specificity of sTREM-1 in diagnosing sepsis were 93. 8％ and 84. 7％, respectively. With a cut-off value of 2. 0 μg/L, the sensitivity and specificity of PCT were 84. 4％and 87. 8％, respectively.ConclusionSerum level of sTREM-1 could be used as an early indicator for diagnosing sepsis.

Clinical study of bacterial infection in patients with liver cirrhosis / 中国医师进修杂志

Objective To study the state, feature and risk factors of bacterial infection in patients with liver cirrhosis, find out the influence of infection on prognosis, and provide scientific basis for its prevention and treatment. Methods Three hundred and twenty-three patients with liver cirrhosis were analyzed. The number of the patients with infection, the location of infection, clinical feature as well as the kind of pathogenic bacteria were analyzed. Unconditional Logistic regression analysis was used to assess the risk factors of bacterial infection. Results The overall infection rate was 39.94% (129/323),of which community acquired infection rate and nosocomial infection rate were 22.60% (73/323) and 17.34%(56/323) respectively. The most common location of infection in turn were respiratory tract,gastrointestinal tract, urinary tract,biliary tract and abdominal cavity. The main pathogenic bacteria was Gram-negative bacillus, most of which had drug resistance for cefquinome and quinolones. The risk factors related with bacterial infection included liver cancer, Child-Pugh class B and C grade of liver function, gastrointestinal tract bleeding, diabetes mellitus,invasive operations and the length of staying in hospital. Conclusions The incidence rate of infection in patients with liver cirrhosis is higher. Multiple factors are likely to affect the incidence rate of infection in patients with liver cirrhosis.

Study on relationships between the expressions of cidA and lrgA and biofilm formation of Staphylococcus epidermidis clinical isolates / 中华微生物学和免疫学杂志

Objective To investigate the relationships between the expressions of cidA and lrgA and the biofilm formation of Staphylococcus epidermidis clinical isolates. Methods Thirty-nine S. epidermidis clinical isolates were divided into biofilm formation group and non-biofilm formation group according to adhesion assays and icaA amplification, 20 strains for biofilm formation group and 19 strains for another group respectively. Amplify cidA and lrgA fragment with PCR, detect mRNA levels of cidA and lrgA with RT-PCR and then calculate the ratios of cid/lrg mRNA. Results All of the 39 isolates of S. epidermidis existed cidA and lrgA gene.Select 2 strains of S. epidermidis, Y36 and Y26, at random for detecting the cidA and lrgA mRNA levels of different phases, the highest level of cidA mRNA shows on 4 h's culture while lrgA shows on6 h. The relative expression levels of cidA of 39 isolates of S. epidermidis were 0.340 + 0.250 in biofilm formation group and 0. 406 +0. 408( P =0. 541 ) in non-biofilm formation group for 4 h culture, while the relative expression levels of lrgA were 0.325 ± 0.218 and 0.253 ± 0.211 respectively ( P = 0. 299). There were no significant differences between these two groups. The ratios of cid/lrg mRNA were 1.067 ±0.529 and 1.958 ±1.877 respectively, the difference of population distribution was significant( P = 0.001 ). Conclusion The expressions of cidA and lrgA of S. epidermidis clinical isolates may be time-limited. There were no significant differeces of expressions of cidA and lrgA between biofilm formation group and non-biofilm formation group. The ratios of cid/lrg mRNA maybe have some biological significance.

The multidrug efflux pump AdeABC in clinical Acinetobacter baumannii isolates / 中华微生物学和免疫学杂志

Objective To study the expression of active efflux pump AdeABC in clinical Acineto-bacter baumannii islates and whether this efflux pump confers resistance to antibiotics. Methods The anti-biotic susceptibility and the function of efflux pump inhibitor were tested by micro-dilution broth method. The expression of adeB was examined by RT-PCR. The controlling gene adeRS was amplified by PCR and se-quenced. Results Thirty multidrug resistance Acinetobacter baumannii isolates and 5 sensitive isolates for PCR were both abtained the expected products of adeB and adeRS. The mRNA expression of adeB in 15 multidrug resistance(MDR) isolates were positive, but there was no expression of adeB in 5 sensitive iso-lates. The mutations of adeRS existed in 2 MDR isolates. Conclusion The expression of AdeABC may in-volved in the resistant mechanisms of the clinical MDR Acinetobacter baumannii isolates.

A Clinical Study of Faropenem on Treatment of Acute Bacterial Infections / 天津医药

Objective: To evaluate the clinical efficacy and safety of domestic faropenem in the treatment of acute bacterial infections. Methods: A multicenter, randomized, double blind and double simulation clinical study was conducted to compare the efficacy and safety of faropenem and cefaclor in the treatment of acute bacterial infection. Patients in trial group(n = 122) were given faropenem 250 mg，and in control group (n = 118) were given cefaclor 200 mg，3 times daily for 7 to 10 days.Results: The clinical cure rates were 33.61% and 27.12% in trail and control groups respectively and the clinical effective rates were 87.70% and 83.05% respectively. There was no significant difference in terms of clinical effectiveness between the two groups（P > 0.05）. The adverse reaction rates were 7.32% in trial group and 3.36% in control group（P > 0.05）. The adverse reaction of the trial group was mainly exaltation of aminotransferase, which did not affect the therapy. No severe adverse reaction was found.Conclusion: Domestic faropenem is effective and safe for the treatment of bacterial respiratory tract and urinary tract infections.

Levels of vascular endothelial growth factor in severe sepsis patients and its relation to prognosis / 中华急诊医学杂志

Objective To investigate the relationship between the serum levels of vascular endothelial growth factor(VEGF) and severity and prognosis by evaluating its changes in severe sepsis patients. Method Us-ing control study design, a total of 29 severe sepsis patients who admired in ICU department of Second Hospital of Tianjin Medical University from July 2006 to November 2007 were enrolled. The patients were divided into survival group (n=16) and death group (n=13) according to the clinical outcomes at 28 days after onset.A total of 31 healthy persons were enrolled into the control group. Clinical and laboratory data including blood routine test,blood-gas analysis,blood chemistry,C-reactive protein,lactic acid were collected on the first,third and 7th day after on-set,respectively. APACHE Ⅱ score were calculated. VEGF levels were determined using ELISA method. Quantita-tive data were analyzed by Ftest. Results The VEGFlevels ofthe control groupwere (78.77±8.15) pg/mL, the VEGF levels of the survival group on the first,third and 7th day were (210.47±59.40) pg/mL, (161.79 ±32.58) pg/mL and (85.33±12.13) pg/mL, respectively. The peak value of VEGF levels appeared on the first day, Then,it decreased with the progression of the disease. The VEGF levels did not differ significantly between the control group and survival group on the 7th day (P>0.05). The VEGF levels in the death group on the first, third and 7th day were (324.12±44.35) pg/mL,(185.40±30.92) pg/mLand (273.32±55.23) pg/mL, respectively. The peak value of VEGF levels in the death group also appeared on the first day, but it did not de-crease significandy on the 7 th day as compared that on theist day. The value of VEGF levels on the 7 th day were significantly higher than that of the control group (P<0.01 ). The levels of VEGF were positive correlated with APACHEⅡ score(rs = 0.510,P<0.01), both VEGF levels and APACHEⅡ score were risk factors for the clinical outcomes of these patients. Conclusions The serum levels of VEGF are elevated at early stage in severe sepsis patients. The VEGF levels, which might be a potential prognositic factor for sepsis patients are significantly correlated with APACHE Ⅱ score.

Analysis of the biofilm formation and icaA genes of Staphylococcus epidermidis isolates / 中华微生物学和免疫学杂志

Objective To determine the ability of biofilm formation of Staphylococcus ephtermidis isolates and analyze the correlation between the icaA gene and its expression and biofilm formation. Methods Collecting 205 Staphylococcus epidermidis isolates identified with normal laboratory tests (coagulase-negative, biochemical identification, polymyxin-resistant and novobiocin-sensitive ), the suspected isolates were con-formed with API-Staph. Biofilm production was assessed by incubating the strains on Congo Red Agar (CRA) plates and quantitative biofilm production determined by a 96-well tissue culture plate and biofilm morphous were detected by scanning electron microscope ( SEM ) ; Amplifying partial fragments of icaA genes with PCR; Analyzing the expression levels of icaA gene with RT-PCR through Bio-Rad system and Quantity One software. Results 24 isolates showed positive in CRA tests, 22 isolates were positive in semiquantita- tive adhesion assays and 28 isolates existed icaA gene among 205 isolates of Staphylococcus epidermidis. The icaA-positive strains demonstrated biofilm formation (microcolonies on silica films ) while icaA-negative strains only adhered as individual cells under scanning electron microscope. All 22 strains which showed positive in semiquantitative adhesion assays harbored the icaA gene. The expression levels of icaA gene with RT-PCR in 6 Staphylococcus epidermidis isolates showed a higher tendency in 4 strains which demonstrated positive in semiquantitative adhesion assays than 2 negative strains in semiquantitative adhesion assays. Conclusion The isolations of Staphylococcus epidermidis have the abilities of forming biofilm, and the icaA gene and its normal expression is the important molecular biology foundation of biofilm formation. Other fac-tors maybe involve in the expression of icaA gene in Staphylococcus epidermidis isolates.

In vitro susceptibility assay of 42 Stenotrophomonas maltophilia with biofilms to antibiotic agents / 中华微生物学和免疫学杂志

Objective To investigate the effects of levofloxacin,ciprofloxacin,ceftazidime,pip-eracillin,cefoperazone/sulbactam,erythromycin,sulfamethoxazole and gentamycin on the bacterial biofilms of Stenotrophomonas mahophilia.Methotis Biofilm and conventional susceptibilities were determined for S.maltophilia isolates from 42 patients.The model of S.maltophilia biofflms in vitro was developed in the Mueller-Hinton broth--micmtiter inoculator or silica films.After antibiotic challenge plate 20 h,each plate was sonicated and the absorbance value at 620 nm(A620)was measured on a microtiter plate colorimeter be-fore and after incubation for 6 h.Then the biofilm inhibitory concentrations were calculated.Finally,based on the acquired data.the experiments of combinafion effects of erythromycin with the 3 antibiotic agents on the formed biofilms of 5 picked strains were designed and worked out.Results The sensitive rate of 42 S.maltaphilia to levofloxacin.sulfamethoxazole and piperacillin were 83.33%,66.67%and 54.76%,re-spectively.The bilfilm inhibitory concentrations were much higher than the corresponding minimal inhibitory concentrateion after formed biofflms.Conclusion Forty-two S.maltophilia are multi-resistant to antibiotic agent.And levofioxacin may have a better effect against biofilms compared with others.The inhibition effect of combination erythromycin with levofloxacin is more obvious among all the 3 antibiotic agents.

Repressor SmeT of Efflux Pump SmeDEF in Stenotrophomonas maltophilia / 中华医院感染学杂志

OBJECTIVE To investigate the repressor SmeT and other sequences of efflux pump SmeDEF in the clinic isolates of Stenotrophomonas maltophilia to involve in antibiotic resistance. METHODS The mRNA level of smeD examined the resistance profile of the clinical strains to antimicrobials was by agar dilution and their reaction to pump inhibitor was investigated by RT-PCR,and the smeD-smeT fragment was amplified by PCR,and then sequenced.Adding poly(A) to 3′ end of RNA,applying 3′RACE,nested PCR,then sequence-analysis. RESULTS The amino acid sequences of the forepart of SmeT were conservative in all the 7 strains.In antibiotic-resistant strains with positive reaction to pump inhibitor,the intergenic sequences of smeD-smeT were different from those in sensitive ones.The variations in(-165 to-82)nt of smeT and several nucleic acids before the start codon of smeT,possibly involved in antimicrobial resistance.All isolates didn′t have Leu166Gln mutation within the SmeT protein of D457R,which was reported possibly associated with antibiotic resistance.The resistant strains with efflux positively inhibited had Asp218Glu substitution,different from the negatively-inhibited ones. CONCLUSIONS The expression level of SmeDEF is related with the antibiotic resistance of S.maltophilia.The possible resistance mechanism includes the variations in the intergenic smeD-smeT and/or smeT codon sequence.

Investigation of the relationship between CTX-M-ESBLs and class Ⅰ integron in enterobacter cloacae clinical strains resistant to third generation cephalosporins / 中华传染病杂志

Objective To study the distribution of CTX-M-ESBLs and class Ⅰ integron in enterobacter cloacae resistant to third generation cephalosporins,and discuss the relationship between CTX-M-ESBLs and class Ⅰ integron. Methods K-B test for resistance;DDST and PCR for CTX-M-(ESBLs);PCR,nested-PCR and sequence analysis for finding class Ⅰ integron carrying CTX-M-ESBLs gene cassette. Results Among 37 strains,there are 21 isolates producing CTX-M-ESBLs,20 isolates with class Ⅰ integron and 13 isolates carrying CTX-M-ESBLs as well as class Ⅰ integron.In 3 isolates: CH4,CH11 and Q1,each class Ⅰ integron haboured CTX-M-ESBLs gene cassette. Conclusions There is few report on class Ⅰ integron carrying CTX-M-ESBLs,which caused more dangers of horizontal transmission of ESBLs and was important factor to nosocomial outbreak of multiresistant clinical isolates.

Gene diversities of ESBLs in clinical isolates of Enterobacteriaceae / 中华传染病杂志

Objective To study the Genotype of local clinical isolates of Enterobacteriaceae producing extended spectrum ? lactamases. Methods K B test, conjugative transfer test, plasmid profile analysis, PCR, PCR RFLP, and DNA sequencing were used to detect the phenotype and genotype of 33 isolates of Enterobacteriaceae producing ESBLs. Results Eighty five percent of 33 isolates was resistant to cefotaxime which was obviously higher than that to ceftazidime. blaCTX M ESBLs was detected in 28 isolates by PCR, blaTEM DNA in 24 isolates, and blaSHV DNA in 9 isolates. Mutation of E104K was only identified in one TEM ? lactamase produced by EC98A7 by PCR RFLP. No substitution of G238S occurred in 9 SHV ? lactamases. DNA sequencing and DNA alignment showed the blaCTX M DNA fragments from 4 clinical isolates of EC98A7,EB56,CFR78 and, KP9941 belonged to CTX M type 1, with highest identity to blaCTX M 3 or blaCTX M 12 respectively. Conclusions CTX M ESBLs is carried in 85% isolates of Enterobacteriaceae producing ESBLs in this city. Most of the isolates carry 2 or more ? lactamases. E. coli EC98A7 produces two ESBLs, a TEM ESBL and a CTX M ESBL.

The mutation of mar gene in clinical isolated Escherichia coli resistant to fluoroquinolones / 中华传染病杂志

Objective To study the antibiotic resistance mediated by the mar mechanism in clinical isolated Escherichia coli. Methods To analyse the antibiotic resistance pattern and the out membrane protein spectrum of clinical isolated E. coli . The plasmid pNJR3 2 carrying the wild type gyr A gene was transferred into clinical multiple resistant strain LC 1. The resistant pattern and plasmid pattern of LC 1 before and after plasmid transferring were compared. The mar OR of clinical strain LC 1 was cloned and sequenced through PCR. The nucleotides of mar gene were compared with the wild type issued by GenBank. Results Of all the 49 clinical strains , six strains express Mrp5, a mar specific out membrane protein. No changes were detected on susceptibility to fluoroquinolone of clinical strain LC 1 after pNJR3 2 transferred suggesting that gyr A was not the main reason contributing to fluoroquinolone resistance. Sequence analysis showed three mutant spots in mar OR of clinical strain LC 1. Conclusion There does exist mar mechanism in clinical isolated Escherichia coli resistant to fluoroquinolone. The mutation in mar OR may contribute to the mar phenotype of LC 1.

RAPD Applied to Molecular Epidemiology of Enterobacter aerogenes in a Neonatal Unit / 中华医院感染学杂志

OBJECTIVE To establish RAPD typing method for Enterobacter aerogenes,and apply RAPD to study molecular epidemiology of E.aerogenes in a neonatal unit.METHODS Five E.aerogenes strains were isolated from four patients in the same neonatal unit at the same time.These strains were typed by RAPD technique.Antibiotic susceptibility was determined by MIC to evaluate drug-resistance.RESULTS Two strains belonging to a unique RAPD-typed ones were epidemiologically related strains.These strains isolated from two patients who hospitalized in the same neonatal unit for four and ten days,respectively.Five E.aerogenes strains were resistant to aminoglycosides,piperacillin and the third-generation cephalosporins in varying degree.CONCLUSIONS RAPD technique is a very easy and reliable molecular tool in the study of E.aerogenes epidemiology.Antibiotic resistance of E.aerogenes is probably related with the history of using antibiotics.