ObjectiveTo observe the clinical efficacy of aligned needling in treating cough in kids.MethodTotally 113 kids with cough were randomized into two groups. Fifty-sixkids in the treatment group were intervened by aligned needling; fifty-seven kids in the control group were by orally takingXiao ErKe Chuan Lingoral fluid.ResultThe total effective rate was 87.5% in the treatment group versus 66.7% in the control group, and the difference was statistically significant (P<0.05).ConclusionAligned needling and orally takingXiao Er Ke Chuan Lingoral fluid both can produce certain efficacies in treating kids’ cough, while the total effective rate of aligned needling is significantly higher.

Objective To study the epidemiology and genotypes of extended-spectrum beta-lactamases (ESBL)-producing Escherichia coli (EC) and Klebsiella pneumoniae (KP) that caused community-onset bloodstream infections (COBSI) in 9 county hospitals of Zhejiang Province.Methods This is a multi-center, prospective, observational study.The cases and isolates with COBSI caused by EC and KP were consecutively collected from 9 county hospitals in Zhejiang Province between 1st March 2014 and 30th April 2015.The double disk diffusion method was used to confirm the production of ESBL.The ESBL genotypes were determined by polymerase chain reaction(PCR) amplification and sequence analysis.Multi-locus Sequence Typing (MLST) was used to analyze the homology of ESBL-producing isolates.Minimal inhibitory concentration (MIC) of frequently used drugs for ESBL-producing isolates was determined by in-vitro antimicrobial susceptibility tests.Results During the study period, a total of 172 cases with COBSI were collected and 171 cases were eligible, among which 126 were caused by EC and 45 were caused by KP.The overall prevalence of ESBL was 34.5% (59/171),and the prevalence of ESBL-EC and-KP was 41.3% (52/126) and 15.6% (7/45), respectively.CTX-M-type ESBL accounted for 96.6% (57/59) of all the ESBLs-producing isolates, and the most common type was CTX-M-14 (27.1%, 16/59), followed by CTX-M-55 (22%, 13/59).MLST analyses revealed significant genetic diversity among ESBL-EC and-KP.The most prevalent ST of ESBL-EC was ST131 (23.1%).In addition to carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam, moxalactam, amikacin and fosfomycin also showed good in-vitro activity against ESBL-EC and-KP.Conclusions The prevalence of ESBL in EC and KP is high in 9 county hospitals of Zhejiang Province, and the most common genotypes are CTX-M-14 and CTX-M-55.The detection rate of ESBL in EC is higher than in KP.It could be considered adequate empirical therapy according to the results of antimicrobial susceptibility tests.Carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam, moxalactam, amikacin and fosfomycin have good in-vitro activity against ESBL-EC and-KP.

Objective To observe the effects of Qutantongluo decoction on the expression of TGF-β1 in renal tissue of diabetic nephropathy(DN) rats,which can help to understand the mechanism of action.Methods The rats models of diabetic nephropathy were established by injecting with streptozotocin intraperi-toneally.All Wistar rats were randomly divided into 6 groups:the normal control group,the model control group,Benazepril Hydrochloridec group ( 1 mg/kg ·d-1 ),Qutantongluo decoction low dose group (3 g/kg ·d-1 ),Qutantongluo decoction median dose group (4.5 g/kg · d-1 ),and Qutantongluo decoction high dose group (6 g/kg · d-1 ).Rats in the normal control group and the model control group were given celiac perfusion of distilled water once at the same time and dose.All the rats were gavaged once daily for 8 weeks.The expression of TGF-β1 at the level of gene and protein in renal tissue were tested by immunohistochemistry and RT-PCR methods.Results Compared with the model control group(2.79±0.22),the expression of TGF-β1 at the level of protein in renal tissue of Benazepril Hydrochloridec group (1.55 ±0.12)and the Qutantongluo decoction low dose group ( 1.54± 0.16),Qutantongluo decoction median dose group (1.49 ± 0.17),Qutantongluo decoction high dose group(l.39±0.25) decreased significantly (P＜0.05).Compared with the model control group (0.35±0.07),the expression of TGF-β1 mRNA in renal tissue of Benazepril Hydrochloridec group(0.35± 0.07)and the Qutantongluo decoction low dose group(0.39±0.03),Qutantongluo decoction median dose group (0.35± 0.06),Qutantongluo decoction high dose group (0.32± 0.07) decreased significantly (P＜0.05).Conclusion Qutantongluo decoction can down regulate the expression of TGF-β1 in kidney,which may be one of the mechanisms of Qutantongluo decoction effectiveness in clinical treatment of diabetic nephropathies.

Objective To evaluate the usefulness of DiversiLab system for genotyping of Acinetobacter baumannii.Methods Fifty-eight non-duplicated clinical Acinetobacter baumannii isolated from 15 cities in China in 2005 were typed by rep-PCR-based DiversiLab system.The results were compared with those of PFGE and multilocus sequence typing. Simpson's index of diversity was used to compare the discriminatory power among DiversiLab system, PFGE and MLST.Results Fifty-eight Acinetobacter baumannii isolates were differentiated into 5 clusters and 25 unique types by DiversiLab system. MLST identified 35 distinct sequence types, which fell into one clone complex of CC22 and 35 singletons, while PFGE resolved 5 pulsotypes and 34 unique types.Simpson's diversity indices for DiversiLab system, MLST and PFGE were O.876, O.944 and 0.961, respectively.Conclusions The discriminatory power of DiversiLab system is lower than that of PFGE and MLST.But as a simple, fast and reproducible typing method, it could be used as a first-line typing tool for the analysis of a large number of isolates.

Objective To construct the mice pneumonia model with imipenem-resistant Acinetobacter baumannii and provide experimental model in anti-pan-resistant Acinetobacter baumannii therapy study. Methods A total number of 120 4-week-old BALB/C male mice were randomly selected and divided into three groups including micro-intratracheal injection, ultrasonic atomizing and nasal dripping. The mice were treated with methotrexate to induce hypo-immunity in every group. These BALB/C mice of normal immunity and hypo-immunity were infected through Imipenem-resistant Acinetobacter baumannii by microintratracheal injection, ultrasonic atomizing and nasal dripping, respectively. The morbidity, mortality,bacterial clearance rate and histopathology in lung were determined. Results The morbidities of BALB/C mice with hypo-immunity infected by micro-intratracheal injection and ultrasonic atomizing achieved 100%(30/30), while the mortalities were 100% (10/10) and 33.3% (3/10), respectively. Mice in two groups above displayed the influx of neutrophils, lymphocytes and macrophages in the peri-bronchial and alveolar interstitial space 12-24 h after pulmonary infection. In addtion, the mice in micro-intratracheal injection group displayed coUapse of partial alveolar walls, formation of abscesses and bacterial colonies in alveoli. While the lung pathology in mice of ultrasonic atomizing group was characterized by cell degeneration in some regions in the lungs, slight relaxation, congestion in alveolar wall vessels and normal of bronchial and alveolar tissue 24 h after inoculation. Degeneration in peri-tracheal and peri-bronchial areas was observed 24-48 h after inoculation, along with highly expanded pulmonary blood vessels and edems. The inflammation was reduced at 48 hours. There was no obvious pulmonary infection in BALB/C mice with hypo-immunity by nasal dripping with mortality of 0% (0/10) and no significant histopathologic change in lungs. Conclusions BALB/C mice with hypo-immunity pneumonia model with Imipenem-resistant Acinetobacter baumannii can be conducted by micro-intratracheal injection or ultrasonic atomizing, but the latter has the advantages of high-productivity, easy-operation, low-cost, time-saving and usefulness. Mice with normal immunity are not susceptible to imipenem-resistant Acinetobacter baumannii.

Objective To investigate the resistant mechanism of carbapenem-resistant Escherichia coli and its relationship with endogenous infection. Methods Two carbapenem-resistant Escherichia coli strains were isolated from blood and stool of a same patient, respectively. The minimal inhibition concentrations (MIC) of the two isolates against imipenem and meropenem were determined by E-test. The susceptibility against other antimicrobial agents were done by disc diffusion method. Isoelectric focusing electrophoresis (IEF), polymerase chain reaction (PCR) amplification,cloning and sequencing, conjugation, Southern blotting were carried out to analyze the encoding gene of β-lactamases. Homology analysis of the two strains was done by pulsed field gel electrophoresis (PFGE). Results MIC against imipenem and meropenem of the two strains were both≥32 mg/L.Both strains produced KPC-2 (pI 6.7) and SHV-12 (pI 8.2) β-lactamases. blaKPC2gene was located on a 54 kb transferable plasmid. PFGE showed that the two Escherichia coli strains were derived from the same clone. Conclusions The resistance and enzyme digestion map of chromosome DNA of the two Escherichia coli strains are coincident. The Escherichia coli septicemia of this patient is probably an endogenous infection caused by the immigration of Escherichia coli from the gut.

Early detection and treatment of high-risk adenomas and colorectal cancer (CRC) can reduce mortality of this disease.CRC screening is aimed at minimizing its harm and colonoscopy is presently the gold standard for it.However,colonoscopy needs bowel preparation and is invasive with high risk of intestinal perforation,causing a bad compliance,which is unfavorable to its popularization and application.Recently,non-invasive detection methods for CRC have gone through a rapid development.Tests based on CRC-related biomarkers in fecal and blood samples provide new options for non-invasive CRC screening.However,detection methods for these biomarkers still need further research and improvement because of the complex composition of feces and blood.In the two aspects of fecal tests and blood tests,the progress of recent studies on non-invasive screening methods for CRC was reviewed in this article.

Objective To evaluate different tigecycline susceptibility testing methods for A.baumannii.Methods Thirty carbapenem resistant A.baumannii (CRAB) and 30 carbapenem sensitive A.baumannii (CSAB) isolates were randomly collected from 30 hospitals during January to December in 2010 in China retrospectively.MIC and inhibitory zone diameters for tigecyclinc were determined by the susceptibility testing methods such as broth microdilution (BMD),agar dilution,E test,MIC Test Strip (MTS),Vitek2.Data were analyzed by comparing the results from each method to those produced by the reference BMD method.The effects of two different susceptibility test media (M-H and ISO-Sensitest Agar) on the MIC of tigecycline were also analyzed.Results For CSAB isolates,the MIC50/MIC90 of BMD,agar dilution,E test,MTS and Vitek2 were as follows:0.125/0.25 mg/L,0.125/0.25 mg/L,0.5/1 mg/L,0.125/0.25 mg/L and 0.5/0.5 mg/L.Compared with BMD method,the categorical agreement rates (CA) of each method were ≥90％,and produced no very major errors (VME) by Food and Drug Administration (FDA)/ European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints.For CRAB isolates,the MIC50/MIC90 of BMD,agar dilution,E test,MTS and Vitek2 were as follows:2/4 mg/L,4/4 mg/L,4/4 mg/L,1/2 mg/L and 2/4 mg/L.Compared with BMD method,MTS produced 3.3％ (1/30)/6.7％ (2/30) VME(FDA/EUCAST breakpoints),and no method CA was ≥90％.The CA between disk diffusion and BMD results were higher by using the criteria of Jones than FDA breakpoints,but only 66.7％ (20/30) were observed in CRAB isolates,and produced no VME.The MIC of tigecycline determined using M-H agar were usually higher than those using ISO-sensitest Agar.Conclusions Agar dilution,E test,Vitek 2 and disk diffusion appear not to be a suitable method for routine susceptibility testing of tigecycline for CRAB strains.Tigecycline intermediate or resistant results determined by these methods require confirmation by BMD,and MTS results also need to be interpreted with caution.

Objective To evaluate the synergy effect of Meropenem and Sulbactam combination against Meropenem-resistant and Meropenem-susceptible A.baumannii in vitro and optimize combination ratio of Meropenem and Sulbactam to achieve best synergy effect.Methods Evaluating the synergy effect of Meropenem and Sulbaetam combination through microdilution checkerboard method against Meropenemresistant and Meropenem-susceptible A.baumannii,isolated from inpatients of Chinese hospitals.Assessing the synergy effect of combination in different ratios of Meropenem to Sulbactam.Results The checkerboard method with the combination of Meropenem and Sulbactam demonstrated 25.0％ ( 10/40 ) synergism,67.5％ (28/40) partial synergism,7.5％ (3/40) additive,no indifference and antagonism in Meropenemsusceptible isolates,and 27.5％ (11/40) synergism,40.0％ (16/40) partial synergism,25.0％(10/40) additive,no indifference and antagonism in Meropenem-resistant isolates.Eleven Meropenemresistant isolates which showed synergism in synergy test were tested for MICs of combination of Meropenem and Sulbactam,using ratios of 4∶ 1,2∶ 1,1∶1 and 1∶2,and the MIC90 were 64∶ 16,64∶ 32,32∶32,32∶64 μg/ml,respectively.Conclusions Meropenem and Sulbactam combination show synergism or partial synergism against most A.baumannii isolates.The optimal ration of combination for clinical use may be 1∶ 1.

Objective To analyze the colistin heteroresistance in Pseudomonas aeruginosa strains and their in vitro susceptibility to antibiotics used in combination.Methods Two hundred and ninety-seven carbapenem-resistant Pseudomonas aeruginosa strains were selected for this study.Broth microdilution method was used to determine the minimum inhibitory concentrations of colistin and other antimicrobials against the Pseudomonas aeruginosa strains.The colistin heterogeneity of 20 colistin sensitive strains was analyzed by using population analysis profiles.The time-kill curves of 3 randomly selected colistin heteroresistant strains were used to determine the bacteriostatic activity of colistin.Chequer-board method was used to measure the combination efficacy of colistin with other antimicrobials including imipenem,meropenem,biapenem,ceftazidime,levofloxcin,piperacillin/tazobactam and cefoperazone/sulbactam.Results The colistin sensitive Pseudomonas aeruginosa strains accounted for 99.66％ of the 297 isolates.Population analysis profiles displayed that 35％ of the 20 isolates were colistin heteroresistant and 20％ of the 20 isolates were heterogeneous.It showed that when colistin was used in combination with other drugs,they mainly had synergistic and additive effects on heteroresistant isolates,but additive and indifferent effects on non-heterogeneous isolates.Conclusion Multidrug resistant Pseudomonas aeruginosa strains were highly susceptible to colistin,but heteroresistant and heterogeneous strains were common.The efficacy of colistin against heteroresistant isolates could be enhanced by using in combination with other drugs.