综合医院病原菌分布及耐药性监测分析

目的：通过对全院住院患者临床分离菌的分布及耐药情况进行监测,及时掌握医院细菌分布和耐药特点。方法：对医院2013年1-12月的临床送检的标本检验结果进行统计分析。采用纸片扩散法或自动化仪器法进行药敏试验,按美国临床实验室标准化协会2013年版标准判读药敏结果,采用WHONET5.6软件进行数据分析。结果：8536株病原菌中革蓝阴性菌占61.95%（5288株）,革蓝阳性菌占31.85%（2719株）,真菌占6.20%（529株）。前三位革蓝阴性菌依次是大肠埃希菌35.21%（1862株）、鲍曼不动杆菌16.72%（884株）和肺炎克雷伯菌13.73%（726株）。前三位革蓝阳性菌依次是金黄色葡萄球菌21.29%（579株）、屎肠球菌21.07%（573株）和表皮葡萄球菌12.91%（351株）;真菌以白色念珠菌为主48.20%（255株）。产ESBLs大肠埃希菌分离率72.0%,产ESBLs肺炎克雷伯菌分离率49.5%,MRSA分离率47.9%,耐碳靑酶烯类的铜绿假单胞菌的分离率31.6%,耐碳靑酶烯类的鲍曼不动杆菌的分离率86.4%,耐万古霉素的屎肠球菌分离率0.5%。呼吸道标本分离出病原菌前三位依次为鲍曼不动杆菌60.73%（532株）、铜绿假单胞菌16.88%（330株）和金黄色葡萄球菌11.20%（219株）;血标本分离出病原菌前三位依次为大肠埃希菌34.43%（498株）、肺炎克雷伯菌10.13%（138株）和表皮葡萄球菌5.43%（74株）;尿标本分离出病原菌前三位依次为大肠埃希菌43.13%（713株）、屎肠球菌12.34%（204株）和肺炎克雷伯菌8.59%（142株）。主要检测的革蓝阳性球菌普遍对万古霉素、利奈唑胺、替考拉宁和替加环素敏感,未发现对万古霉素、替考拉宁、替甲环素和利奈唑胺耐药的金黄色葡萄球菌,发现少数万古霉素和替考拉宁耐药的屎肠球菌和表皮葡萄球菌及少数对利奈唑胺和替加环素耐药的粪肠球菌;除了鲍曼不动杆菌外,其他检测的革蓝阴性杆菌对碳青酶烯类仍然最敏感。结论：细菌耐药性仍是临床重要问题,尤其是泛耐药鲍曼不动杆菌,应加强监测,指导临床合理使用抗生素。

Screening of Chinese Herbal Medicines Resistant to Chicken Escherichia coli and Infectious Laryngotracheitis Virus

[Objective] This study aimed to screen Chinese herbal medicines resistant to Chicken Escherichia coli and infectious laryngotracheitis virus. [Methed] Conven- tional punch method, test tube method and plate dilution method were adopted for in vitro susceptibility test of chicken E, coil strains O5 and O8 using 13 kinds of Chi- nese herbal medicines including Sanguisorba officinalis, Coptis chinensis, Anemar- rhena asphodeloides, Strobilanthes cusia, Agastache rugosa, etc.; chicken embryo inoculation experiment was adopted to screen Chinese herbal medicines resistant to chicken infectious laryngotracheitis virus. [Result] Sanguisorba officinalis, Fructus mume, Rheum officinale, Coptis chinensis, Herba Taraxaci, Anemarrhena asphode- Ioides, Scutellaria baicalensis and Rhizoma Fagopyri Cymosi had ideal antibacterial effect against chicken E. coil strain O5; Sanguisorba officinalis, Fructus mume, Rheum officinale, Coptis chinensis, Herba taraxaci and Rhizoma Fagopyri Cymosi had ideal antibacterial effect against chicken E. coil strain 08; other Chinese herbal medicines showed relatively poor or no antibacterial effect. Results of chicken embryo inoculation experiment showed that nine kinds of Chinese herbal medicines showed relatively strong anti-lLTV effect, including Forsythia suspensa, Radix Isatidis, Fofium isatidis, Flos Ionicerae, Radix codonopsis, Radix astragali, Atractylodes, Radix gly- cyrrhizae, and Pericarpium granati. [Conclusion] The study laid the foundation for fur- ther development of Chinese herbal compound preparations to treat chicken cofibacil- Iosis, infectious laryngotracheitis and other bacterial, viral diseases.

Analysis of Culture and Drug Sensitivity Tests of Mycoplasmas for 387 Patients with Nongonococcal Urethritis (Cervicitis) in Chongqing

Objective: To investigate the prevalence of myco-plasma infections and the sensitivity to antibiotics among patients with nongonococcal urethritis or cer-vicitis (NGU) in Chongqing. Methods: 387 NGU cases with mycoplasma-positive results upon culture were analysed retrospectively. RESULTS: The majority of patients with mycoplasma infections were in the 20-40 year old age group. No significant difference was found between males and females. Ureaplasma urealyticum is the main pathogen of these NGU cases and no clear relationship between its concentration and pathogenic ability was noted. Drug sensitivity was tested against nine antibiotics; the sensitivity rates to josamycim, minocycline and doxycycline were 94.06%, 88.89% and 86.82% respectively, while the resistance rates to lincomycin, ofloxacin, azithromycin and roxthromycin were 74.94%, 42.12%, 41.60% and 40.31% in turn. Conclusions: Josamycin, minocycline and doxycycline could be used as the first choice to treat NGU with mycoplasma infections in Chongqing. It is important to select antibiotics for NGU treatment with mycoplasma infections based on the results of drug sensitivity tests.

Diagnosis and Treatment of Hemorrhagic Pneumonia of Mink

[Objective] The paper was to diagnose and treat hemorrhagic pneumonia of mink. [Method] The diseased minks bred by one raiser in Huangtian Village of Changli County in Qinhuangdao City were used as experimental materials. It was necropsied, and its tissues such as liver, kidney, spleen and lung were taken under aseptic condition, then inoculated into general nutrition agar and blood agar. The bacteria were isolated and purified, and the strains obtained were carried out animal inoculation test and drug sensitivity test. Finally, the relevant treatment and preven- tive measures were put forward. [Conclusion] The bacteria isolated were detected to be negative by Gram staining; it formed hemolysis ring in blood agar; the bacteria could ferment glucose and xylose, which could not ferment sucrose, lactose, maltose and esculin; it was positive in catalase and oxidase test, and negative in urease test and hydrogen sulfide test; so the strain was indentified to be Pseudomonas aerugi- nosa. Drug sensitivity test showed that the bacteria were highly sensitive to ciprofloxacin, which had tolerance to cefoxitin. [Conclusion] The study laid foundation for diagnosis and treatment of hemorrhagic pneumonia of mink.

A case of advanced intrahepatic cholangiocarcinoma successfully treated with chemosensitivity test-guided systemic chemotherapy

Intrahepatic cholangiocarcinoma （ICC） is a relatively rare and highly fatal neoplasm that arises from the biliary epithelium. Prognosis is generally poor and survival is limited to a few months. Here we present a case of advanced ICC successfully treated by chemosensitivity test-guided systemic chemotherapy combining S-1 and cisplatin （CDDP）. A 65-year-old woman with a liver tumor was referred to our hospital on November 21, 2007. Abdominal ultrasonography and computed tomography （CT） showed low-density masses of 50 and 15 mm in diameter, respectively in segment w of the liver and in the enlarged lymph node in the para-aorta. Ultrasonography-guided fine needle biopsy diagnosed the tumors as ICC. Since the patient was inoperable for lymph node metastasis, she underwent systemic chemotherapy with gemcitabine. Six months afcer initiation of chemotherapy, CT revealed ICC progression in the liver and pleural dissemination with pleural effusion. The patient was admitted to our hospital for anticancer drug sensitivity testing on June 9, 2008. Based on the sensitivity test results, we elected to administer systemic chemotherapy combining S-1 and CDDP. Two months into the second chemotherapy treatment, CT revealed a reduction of the tumors in the liver and lymph node and a decrease in pleural effusion.After eight cycles of the second chemotherapy, 17 mo after ICC diagnosis, she is alive and well with no sign of recurrence. We conclude that chemosensitivity testing may effectively determine the appropriate chemotherapy regimen for advanced ICC.

Isolation,identification,and antifungal susceptibility test for Kodamaea ohmeri:a case report on endocarditis

A 43-year-old man with a history of rheumatoid heart disease developed endocarditis.Blood culture showed endocarditis was caused by Kodamaea ohmeri and the susceptibility test showed the yeast species were susceptible to itraconazole,amphotericin B,and voriconazole,but susceptible-dose dependent to fluconazole,and resistant to 5-flucytosine.Treated with surgery and anti-fungi agents,the patient recovered from endocarditis.This is the first case of K.ohmeri fungemia found in Chinese from mainland.More and more evidence indicate that K.ohmeri is an important opportunistic pathogen for human beings.

Isolation and Identification of Drug-resistant Escherichia coli Strains from Chickens

[Objective] This study aimed to isolate and identify the drug-resistant Es- cherichia coli strains from chickens. [Method] E. coli strains were isolated from the fecal samples collected from five chicken farms around Shangqiu City, and verified by biochemical and pathogenic assay. [Result] Among the 35 isolated E. coli stains, 11 E. coil stains were sensitive to florfenicol, amikacin, neomycin and gentamicin; 12 E. coli stains were moderately sensitive to ciprofloxacin, doxycycline and norfloxacin; 15 E. coil stains were resistant against erythromycin, penicillin and streptomycin. [Conclusion] Strengthening biosecurity measures, rationally using vaccine and choosing effective antibiotics are the most cost-efficient methods to control E. coli.

Identification of eight genes that are potentially involved in tamoxifen sensitivity in breast cancer cells

Although the antiestrogen agent tamoxifen has long been used to treat women with hormone receptor positive inva-sive breast carcinoma, the mechanisms of its action and acquired resistance to tamoxifen during treatment are largelyunknown. A number of studies have revealed that over-activation of some signaling pathways can cause tamoxifenresistance; however, very little information is available regarding the genes whose loss-of-function alternation contrib-ute to tamoxifen resistance. Here we used a forward genetic approach in vitro to generate tamoxifen resistant cells fromthe tamoxifen sensitive breast cancer cell line ZR-75-1, and further identified the disrupted gene in different tamoxifenresistant clones. Retinol binding protein 7, DNA polymerase-transactivated protein 3, γ-glutamyltransferase-like activity 1,slit-robo RhoGTPase-activating protein, tetraspan NET-4, HSPC194, amiloride-sensitive epithelial sodium channel gene,and Notch2, were the eight mutated genes identified in different tamoxifen resistant clones, suggesting their requirementfor tamoxifen sensitivity in ZR-75-1 cells. Since the functions of these genes are not related to each other, it suggeststhat multiple pathways can influence tamoxifen sensitivity in breast cancer cells.

Impact of MTT based tumor chemosensitivity assay in vitro

Objective： The aim of the study was to evaluate the impact of the chemosensitivity assay in vitro and establish a standard process of measuring the anti-cancer drug sensitivity with MTT assay. Methods： Some influencing factors of MTT assay in studying the sensitivity of human peripheral blood mononuclear cells （PBMC） to anti-cancer drugs were observed, including red blood cells, platelets, three different kinds of DMSO and different concentrations of MTT. Meanwhile the stability of tumor drug-coated plate was monitored. Results： The red blood cells and platelets may affect the results at a certain range of concentration. Analytical pure DMSO, both imported and domestic reagents showed the same color with MI3-, and the A values of the reaction were dependent on MTT dose. The stability of the freeze-drying drug-coated plates was superior to non freeze-drying ones. Conclusion： To make clear and definite all kinds of influencing factors might contribute to a kind of standard MTT assay for drug sensitivity test in vitro.

Application value of ATP based bioluminescence tumor chemosensitivity assay in the chemotherapy for hydrothorax caused by non-small cell lung cancer

Objective： The aim of the study was to investigate the clinical value and application of ATP based bioluminescence tumor chemosensitivity assay （ATP-TCA） in the chemotherapy for hydrothorax caused by non-small cell lung cancer （NSCLC）. Methods： Hydrothorax specimens from 120 NSCLC patients were analyzed by ATP-TCA and the most sensitive chemotherapeutic drugs were used in NSCLC patients （treatment group）. At the same time, 56 NSCLC patients with hydrethorax were admitted in our Hospital （Department of Oncology, The No. 2 People＇s Hospital of Yibin, China） and given chemotherapy without guidance of the ATP-TCA （control group）. Before the third chemotherapeutic cycle, clinical outcomes were analyzed in the two groups. Results： Effective rate of hydrothorax in treatment group was 67%, while 46% in control group （P 〈 0.05）. In refractory hydrothorax patients, they were 69% and 40% （P 〈 0.05）, respectively.In vitro results correlated well with clinical outcomes （P 〈 0.01）. Conclusion： Effective rate of chemotherapy for hydrothorax in NSCLC is higher in treatment group than that in control group. ATP-TCA is especially helpful for refractory hydrothorax.

Empirical Study on Chemotherapeutic Susceptibility of SP Cells in Human Pulmonary Adenocarcinoma Cell Line A549 in Vitro

OBJECTIVE To explore the chemotherapeutic susceptibility of SP cells in human pulmonary adenocarcinoma cell line A549 and the possible mechanism of multidrug resistance.METHODS SP and non-SP （NSP） cells in the cell line A549 were isolated by fluorescence activated cell sorter. The susceptibility of SP and NSP cells to DDP, 5-FU, VP16, NVB and GEM was detected by a drug susceptibility test, and IC50s were calculated 24 h a er the chemotherapy; and then a er a 2-hour IC50 treatment with 5 chemotherapeutic drugs on the 2 subsets of NSP cells, the intracellular drug levels were determined and analyzed using high performance liquid chromatograph.RESULTS There was no statistical signifi cance in comparison of the di. erences in IC50s and in intracellular drug levels a er DDP treatment between the 2 subsets （P 〉 0.05）, （P 〉 0.05）. However,all IC50s of the other 4 drugs were significantly higher in the SP cells than in the NSP cells （P 〈 0.01）. A er the chemotherapy, the intracellular drug levels of the other 4 drugs were significantly lower in SP cells than in NSP cells （P 〈 0.01）.CONCLUSION Compared to NSP cells, SP cells in the cell line A549 have stronger resistance to the chemotherapeutics. The multidrug resistance of SP cells closely correlates with the function of SP cells discharging chemotherapeutic agents.

Prevalence, genetic diversity, and antimicrobial susceptibility profiles of Staphylococcus aureus isolated from bovine mastitis in Zhejiang Province, China

This study was conducted to determine genetic diversity and antimicrobial susceptibility profiles of Staphylococcus aureus recovered from bovine mastiffs in Zhejiang Province, China. Out of 3178 quarter milk samples from 846 lactating cows, among which 459 cows （54.3%） were found HMT positive, 890 quarters （28%） were found having subclinical mastitis. From 75 representative S. aureus isolates, 16 distinct types were identified by pulsed-field gel electrophoresis （PFGE）. Four major PFGE types （A, B, C, and D） accounted for 82.7% of all isolates, and type A （41.3%） was observed in multiple herds across the studied areas. Each region was found to have a predominant type： Hangzhou type A （64.1%）, Ningbo type C （34.5%） and type B （23.1%）, Jinhua type D （53.3%）, and Taizhou type C （62.5%）. Results of antimicrobial susceptibility tests showed that 90.7% of the isolates were resistant to at least one antimicrobial. Resistance to penicillin and ampicillin （77.3%）, tetracycline （60.0%）, or erythromycin （48.0%） was observed. The bacteria resistant to multiple antibiotics such as penicillin, ampicillin, tetracycline, and erythromycin were commonly found. The information obtained from this study is useful for designing specific control programs for bovine S. aureus mastitis in this region.

Investigation and analysis of the characteristics and drug sensitivity of bacteria in skin ulcer infections

Purpose： Skin ulcer is a common type of disease affecting patients＇ health and quality of life, and bacterial infection increases the difficulty of its management. Methods： The present study collected the results of bacterial culture sampled from the surface of 110 cases of skin ulcers at our hospital from January 2011 to December 2012. We analyzed the constituent ratios of ulcer surface bacteria, the change in the main infectious bacteria and the results of drugsensitivity testing for common bacteria. In addition, the characteristics of bacterial infection of skin ulcers were summarized. Result： Of the 110 samples, 90 isolated bacteria were cultured. Sixty-one were Gram-negative bacteria, mainly comprising Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter cloacae and Escherichia coli. In addition, 23 isolates were Gram-positive bacteria, mainly comprising Staphylococcus aureus and Enterococcus faecalis. The probability of a negative bacterial culture in 2012 was significantly lower than that in 2011 （16.7% vs. 40.0%, p 〈 0.01）. Moreover, the probability of P. aeruginosa infection in 2012 was significantly higher than that in 2011 （31.7% vs. 14.0%, p 〈 0.01）. P. aeruginosa was resistant to seven commonly used antibiotics. Both K. pneumoniae and E. coli had higher resistance to ampicillin. E. cloacae were not sensitive to piperacillin/tazobactam. Acinetobacter baumannii was resistant to all the tested drugs. S. aureus, E. faecalis and Staphylococcus epidermidis had high resistance to clindamycin. There was other drug resistance to reflect the higher rate of skin bacterial resistance. Conclusion： Skin bacterial resistance rate is high. Gram-negative bacteria gradually account for the majority, and P. aeruginosa becomes the most important skin infection pathogen. These characteristics of bacterial infections of skin ulcers provide a significant reference for guiding the selection of antibiotics, better controlling infections of skin ulcers and accelerating the healing of skin ulcers.