目的 了解我院肺炎链球菌的临床分布及耐药情况,为临床合理应用抗菌药物提供依据。方法 采用WHONET 5.6软件对我院2012年—2016年培养、分离和鉴定出的肺炎链球菌的临床分布及药敏试验结果进行分析。结果 2012年—2016年共检出肺炎链球菌519株(不含重复菌株),每年秋冬和初春季节检出率最高。五年检出的肺炎链球菌对各类抗生素的耐药率变化不大。从8个科室和病区分离出此菌,以呼吸科为主,分离出313株,占60.3%。痰液中共分离出488株,占94.03%,其次从血液中分离出32株,占4.24%。对抗生素耐药率大于60%的有:复方新诺明、四环素、克林霉素和红霉素;未出现耐药的抗生素有厄他培南、莫西沙星、利奈唑胺和万古霉素;其余抗生素的耐药率均小于30%,其中肺炎链球菌对青霉素的耐药率为2%、中介率为20%。结论 青霉素仍可以作为治疗肺炎链球菌感染的首先药物;三代头孢菌素可用于青霉素非敏感的肺炎链球菌(PNSSP)治疗;未发现万古霉素非敏感菌株;红霉素、克林霉素的耐药率极高,不适合肺炎链球菌的治疗。临床应当根据培养药敏结果合理使用抗菌药物,减少细菌耐药率的发生。

青蒿素类药物作为抗疟特效药,其特殊分子骨架和过氧基在抗疟中起着关键作用。通过损伤虫体的器膜、诱导蛋白变性、抑制ATP 蛋白6(Pf ATP6)活性等方式杀灭疟原虫。恶性疟原虫基因序列的突变和长期不规范用药均会使其产生耐药性。本文依据文献报道,对世界关于青蒿素类药物抗疟机制、耐药性的产生原因作一综述。

Artemisinin is an effective anti-malaria drug, It's special molecular framework and peroxy group play a key role in antimalarial. Plasmodium falciparum was killed by the inducing cytoplasmic organelledamage protein denaturation, inhibiting ATP6 activity. Mutations in the genetic sequence of plasmodium falciparum and long-time using drugs without rule will develop drug resistance. This article is based on the literature, do a review of the world's causes for the resistance of artemisinin to antimalarial mechanisms.

目的 分析2011—2016年间铜绿假单胞菌分离株的耐药性及变迁情况, 为临床合理用药提供科学依据。方法 对2011年1月—2016年12月广州市第一人民院患者各类标本中分离到的铜绿假单胞菌2 257株进行细菌鉴定及药敏试验,并对耐药性变迁进行统计分析。结果 铜绿假单胞菌在痰液标本中的检出率最高为56.9％;6年铜绿假单胞菌平均耐药率以妥布霉素最低,为9.9%,对哌拉西林/他唑巴坦、头孢吡肟、头孢他啶、左氧氟沙星、环丙沙星、亚胺培南、庆大霉素等药物的耐药率均＜20%,在2013年耐药率最低,此后三年逐年上升。结论 铜绿假单胞菌对广州市第一人民院常用抗生素的耐药率在近3年呈逐年上升趋势, 临床医师应根据药敏结果合理选择抗菌药物, 以提高疗效和减缓耐药菌的产生。

Objective To analyze the changes of drug resistance of Pseudomonas aeruginosa (Pae) and to provide basis for the use of antibiotics in clinic. Methods 2 257 strains of Pae were cultured and isolated in the First People Hospitalof Guangzhou from 2011 to 2016, API bacterial identification system was applied to carry out bacterial identification and K-B method was used for drug sensitivity analysis. Results Most of the Pae (56.9％) were detected from the sputum specimen. It showed the highest sensitivity to tobramycin. The drug resistance of Pae to piperacillin/tazobactam, cefepime, ceftazidime, levofloxacin, ciprofloxacin, imipenem and gentamicin in 2013 was the lowest and has been increasing year by year. Conclusion Pseudomonas aeruginosa isolated in our hospital showed a rising trend of clinical drug resistance in the past three years. It was of the top priority for clinicians to use antibiotics rationally to retard the production of drug resistant strains.

目的 了解广东地区糖尿病足患者创面病原菌分布及耐药性变迁。方法 回顾性分析A组(2010年1月—2014年12月就诊的糖尿病足患者)和B组(2015年1月—2018年1月就诊的糖尿病足患者)研究者创面病原菌分布及耐药性变迁。结果 B组中创面G⁺菌及G^-菌均有下降趋势且G^-菌下降较快,细菌种类明显增加,真菌及混合感染明显上升,A组以金黄色葡萄球菌、铜绿假单胞菌、大肠埃希菌感染为主;B组以金黄色葡萄球菌、奇异变形杆菌、铜绿假单胞菌及真菌感染为主;B组相对于A组的细菌耐药性增加。结论 近年来糖尿病足患者病原菌种类明显增加且混合感染及真菌感染上升,且其耐药性增加,因此早期经验用药而后根据药敏选择抗菌药物治疗是糖尿病足感染治疗的关键。

Objective To investigate the distribution and drug resistance of pathogenic bacteria in diabetic foot wounds in Guangdong area. Methods Patients with diagnosis of diabetic foot between group A (from Jan 2010~Dec 2014 ) and group B (from Jan 2015 to Jan 2018) were retrospectively analyzed. We studied the bacteria distribution and drug resistance of pathogenic changes of group A and group B. Results In group B, both G+ and G- bacteria had a decreased trend while G- bacteria decreased rapidly, but the species of bacteria increased obviously just as fungi and mixed infection increased obviously. Bacteria infection in group A were mainly about Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli wihle group B were Staphylococcus aureus, Proteus mirabilis, Aeruginosa and Fungal infection; the resistance of group B to A was lower. Conclusion In recent years, kinds of pathogenic bacteria of diabetic foot were significantly increased and the mixed infection and increased fungal infection as well as its drug resistance increased, so the early experience of medication choice of antibiotics based on drug sensitivity and treatment are the key to the treatment of diabetic foot infection.

目的 通过高通量测序法对多重耐药大肠埃希菌HX43进行耐药分子机制的研究。方法 用Illumina Miseq平台对HX43进行高通量测序,用Edena、RAST、ResFinder、MLST和BLAST等生物信息学工具或数据库进行数据分析,获得耐药基因相关序列数据。结果 HX43对多种临床常用抗生素均不敏感,仅对碳氢霉烯类药物敏感。对高通量测序数据的分析研究发现,该菌存在多种耐药基因,包括β-内酰胺类耐药基因3个(bla_CMY-42、bla_CTX-M-14和bla_OXA-30),氨基糖苷类耐药基因5个(aac(3)-IIa、aadA5、 strA、 strB和aac(6′)-Ib-cr),喹诺酮类耐药基因1个(aac(6′)-Ib-cr),磺胺及甲氧苄啶类耐药基因3个(sul1、sul2和dfrA17),四环素耐药基因1个(tet(B)),氯霉素耐药基因2个(catB3和cmlA1),大环内酯类耐药基因2个(erm(B)和mph(A))。对包含bla_CMY-42的contigs进行分析,发现该基因与ISEcp1插入序列、blc和sugE等基因相关联。质粒分型发现HX43携带5种不相容群的质粒。多位点序列分型(MLST)分析发现HX43属于ST3835,为国内外较少见的序列型。结论 高通量测序技术可准确获得临床菌株抗生素耐药的相关基因信息,为临床抗菌治疗提供重要的实验室数据支持。

Objective To investigate the molecular resistance mechanism of Escherichia coli HX43 by high-throughput sequencing. Methods HX43 was sequenced by the Illumina Miseq platform, and sequencing data were analyzed by the Edena, RAST, ResFinder, MLST and BLAST softwares and databases. Results HX43 was resistant to most common clinical antibiotics except carbapenems. Analysis of data revealed resistance genes to β-lactams (bla_CMY-42, bla_CTX-M-14 and bla_OXA-30), aminoglycosides (aac(3)-IIa, aadA5, strA, strB and aac(6′)-Ib-cr), quinolones (aac(6′)-Ib-cr), trimethoprim/sulfonamides(sul1, sul2 and dfrA17), tetracyclines (tet(B)), chloramphenicol (catB3 and cmlA1), macrolides(erm(B) and mph(A)). Sequence analysis of the contig containing bla_CMY-42 identified correlations of the gene with ISEcp1 insertion sequences, blc and sugE genes. Plasmid typing identified 5 plasmid incompatibility groups in HX43. MLST analysis found that HX43 belonged to ST3835, a relatively rare sequence type in the world. Conclusion Information of resistance genes can be obtained by high-throughput sequencing, which provides important experimental data for clinical antimicrobial treatment.

目的 研究雷公藤红素对人阿霉素耐药MCF-7/ADR乳腺癌细胞生长的作用。方法 采用MTT试验检测MCF-7/ADR细胞对阿霉素的耐药情况以及雷公藤红素对MCF-7/ADR细胞生长的影响;采用Annexin V-FITC/PI双染试验分析雷公藤红素对MCF-7/ADR耐药细胞凋亡的诱导作用;应用流式细胞周期分析检测雷公藤红素对MCF-7/ADR耐药细胞周期的影响。结果 MCF-7/ADR细胞对阿霉素耐药指数达14.54;而雷公藤红素能有效抑制阿霉素耐药细胞MCF-7/ADR的生长,并呈现浓度依赖性,作用48 h的IC50是1.04 μmol/L,MCF-7/ADR对雷公藤红素的耐药指数仅为0.87。2 μmol/L雷公藤红素作用8 h后,Annexin V-FITC染色阳性的MCF-7/ADR细胞比例较对照显著升高,差异有统计学意义(P＜0.05)。在1 μmol/L雷公藤红素作用24 h后,G1期细胞比例由对照(59.22±3.78)%升高至(77.44±4.21)%,而S期细胞比例由对照(37.51±2.91)%降至(19.65±2.25)%,差异有统计学意义(P＜0.05)。结论 雷公藤红素能激活MCF-7/ADR细胞凋亡的发生,并诱导MCF-7/ADR发生 G1/S细胞周期阻滞,从而对阿霉素耐药MCF-7/ADR细胞的生长发挥高效抑制作用。

Objective To investigate the effect of celastrol on the growth of adriamycin-resistant MCF-7/ADR breast cancer cells. Methods The resistance situation of MCF-7/ADR cells to adriamycin and the effect of celastrol on the growth of adriamycin-resistant MCF-7/ADR cells were evaluated by MTT assay. The effect of celastrol on apoptosis in MCF-7/ADR breast cancer cells was determined by annexin V-FITC/PI double staining. The effect of celastrol on cell cycle progression was determined by flow cytometry analysis. Results The resistance index of MCF-7/ADR cells to adriamycin was 14.54. After treatment with celastrol for 48 h, MCF-7/ADR cells displayed markedly inhibited growth in a dose-dependent manner, and calculated IC50 was 1.04 μmol/L. Celastrol decreased the resistance index of MCF-7/ADR from 14.54 to 0.87. The numbers of apoptotic MCF-7/ADR cells, as revealed by annexin V binding, significantly increased upon celastrol treatment (P＜0.05). Celastrol treatment caused an increase of cells in G1 phase from (59.22±3.78)% to (77.44±4.21)%, while the percentage of cells in S phase was decreased from(37.51±2.91)% to(19.65±2.25)%(P＜0.05). Conclusion These data demonstrated that celastrol induced apoptosis and G1/S cell cycle arrest in MCF-7/ADR cells and consequently displayed potent cytocidal effect on adriamycin-resistant MCF-7/ADR breast cancer cells.

目的 了解龙川地区肺炎支原体耐药情况,以便临床合理运用抗生素。方法 收集2014年—2015年间疑似肺炎支原体感染住院小儿患者的咽分泌物标本2 666例,同时作Mp培养及IgM检查,同为阳性者共149例进行耐药性统计分析。结果 在2 666例疑似感染患者中,培养肺炎支原体阳性149例,阳性率为5.59%,学龄前儿童(≤3岁)患者阳性率2.06%,学龄儿童(4～15岁)阳性率9.79% ,学龄儿童感染肺炎支原体与学龄前儿童比较有显著差异(P<0.05)。其中红霉素、阿奇霉素、罗红霉素、克林霉素、依托红霉素、克拉霉素、乙酰螺旋霉、交沙霉素、加替沙星、莫西沙星、环丙沙星、左氧氟沙星、多西环素、米诺环素耐药率分别为2%、12.4%、4%、22% 、2%、1%、80%、44%、2%、15%、10%、1%、61%、44%。冬季感染与在其他季节比较有差异(P<0.05)。结论 阿奇霉素,红霉素为代表大环内脂类抗生素仍可以作为临床一线经验用药,乙酰螺旋霉、交沙霉素耐药率大于40%,不建议作为经验药物使用。喹诺酮类抗生素耐药率一般小于15%,可作为肺炎支原体治疗的第二选择。四环素类抗生素在本地区耐药性高,不建议作为经验药物使用。要加强预防冬季肺炎支原体感染。

目的 构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型。方法 采用低浓度加量持续诱导法,诱导吉西他滨耐药乳腺癌细胞4T1耐药株,命名为4T1/Gem;CCK-8法测定4T1与4T1/Gem细胞的增殖抑制率,计算耐药指数; Western blot法检测细胞P-gp蛋白表达;B超引导下注射4T1/Gem细胞悬液诱导裸鼠肝脏成瘤;HE染色观察肿瘤组织病理情况,免疫组化法检测瘤组织ER、PR、HER2、Ki-67和P-gp蛋白的表达。结果 经过14个月的诱导成功建立4T1/Gem细胞株,可在含40 μg/mL的Gem培养液中稳定生长。4T1/Gem细胞耐药指数为4T1细胞的788.547倍。与亲代相比,4T1/Gem处于G1期和G2期的细胞增加,S期细胞减少;上调P-gp蛋白的表达。4T1/Gem细胞成功建立裸鼠乳腺癌肝转移模型,瘤组织中ER、PR、HER2蛋白阴性表达,Ki-67阳性10％和P-gp蛋白阳性表达。结论 成功构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型,为开发治疗乳腺癌肝转移化疗耐药的药物提供实验基础。

Objective To construct a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem) and establish a nude mouse model of breast cancer with hepatic metastatic. Methods A gemcitabine-resistant variant of the breast cancer 4T1 cell line was induced by gradually increasing the concentration of gemcitabine; this variant is referred to in this study as 4T1/Gem. The proliferation suppression rates of 4T1 and 4T1/Gem cells were determined by using the CCK-8 essay to evaluate the drug resistance indices of the cell lines. Western blot analysis was used to detect P-gp protein expression. Under ultrasonography, a 4T1/Gem cell suspension was injected into nude mice to induce liver tumors. H&E staining was used to observe tumor pathology, and immunohistochemistry was used to detect the expression of ER, PR, HER-2, Ki-67, and P-gp. Results After 14 months of induction, a 4T1/Gem cell line is established successfully. The cell line can grow stably in culture liquid containing 40 μg/ml gemcitabine. The drug resistance index of 4T1/Gem is 788.547. Compared with the 4T1 cell line, the 4T1/Gem cell line can upregulate P-gp protein expression and successfully establish a nude mouse model of breast cancer with hepatic metastatic. ER, PR, and HER-2 proteins exhibit negative expression in the tumor tissue. The positive expression of P-gp and 10% of Ki-67 proteins is also observed. Conclusion This study successfully constructs a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem)and establishes a nude mouse model of breast cancer with hepatic metastatic, thereby providing an experimental basis for developing and treating a drug-resistant variant of breast cancer.

目的 分析我院2011—2015年我院儿科住院患者下呼吸道病原菌分布及其耐药性。方法 采用全自动生化鉴定仪对痰标本分离株进行鉴定,用全自动微生物药敏系统和纸片扩散法对病原菌的耐药性进行检测,并用头孢硝噻吩纸片法对β-内酰胺酶进行检测。结果 2011—2015年共分离得到下呼吸道病原菌518株,包括肺炎链球菌(21.62%)、金黄色葡萄球菌(16.99%)、流感嗜血杆菌(14.48%)、肺炎克雷伯菌(11.97%)、大肠埃希菌(8.11%)、卡他莫拉菌(5.41%)、鲍曼不动杆菌(3.86%)和铜绿假单胞菌(3.86%)等。药敏结果显示,肺炎链球菌对克林霉素(90.18%)、红霉素(92.86%)和复方新诺明(87.50%)的耐药率较高,金黄色葡萄球菌则对青霉素G(90.91%)和红霉素(68.18%)有较强耐药性,未发现对万古霉素或利奈唑胺耐药的革兰阳性球菌。流感嗜血杆菌对氨苄西林耐药率为32%,与其β-内酰胺酶阳性率较一致,肺炎克雷伯菌和大肠埃希菌对头孢类药物(17.33%~45.33%)和喹诺酮类药物(34.67%~50.67%)耐药性较高,并发现1株碳青霉烯耐药的肺炎克雷伯菌。结论 本院下呼吸道感染病原菌谱较广,主要包括多种革兰阳性球菌和革兰阴性杆菌,并对多种抗菌药物表现出较强耐药性,临床应注重合理应用相关抗生素,严格防控病原菌的医院感染及传播。

Objective To analyze the antimicrobial resistance and the profile of pathogens from lower respiratory tract infections in pediatric in patients. Methods Sputum bacterial isolates were identified by an automated biochemical identification system. Antimicrobial resistance was detected by an automated drug susceptibility detection system and the disc diffusion method. The β-lactamases was tested by the nitrocefin disc detection method. Results Five hundred and eighteen bacterial pathogens were isolated from sputum samples during 2011-2015, including streptococcus pneumoniae(21.62%), staphylococcus aureus(16.99%), haemophillus influenzae(14.48%), klebsiella pneumoniae(11.97%), escherichia coli(8.11%), moraxelle catarrhalis(3.8%), acinetobacter baumanii(3.86%) and pseudomonas aeruginosa(3.86%). High resistant rates were detected for S. pneumoniae to clindamycin(90.18%), erythromycin(92.86%) and sulfamethoxazole (85.50%), while S. aureus was highly resistant to penicillin G(90.91%) and erythromycin(68.18%). Resistance to vancomycin and linezolid was not detected for gram positive cocci. The resistant rate to ampicillin was 32% for H. influenzae, which was in concordance with the production of β-lactamases. Relatively high resistance was detected for K. pneumoniae and E. coli to cephalosporins and quinolones. A carbapenem-resistant K. pneumoniae isolate was also detected. Conclusion Multiple bacterial species were isolated from lower respiratory tract infections in our hospital, including different species of gram positive cocci and gram negative bacilli, and these isolates exhibited high resistance to antibiotics tested. The clinical use of antibiotics and hospital infection and transmission of these pathogens should be controlled.

目的 了解中山市7家医院金黄色葡萄球菌感染的临床分布,并对耐药基因进行检测,为临床经验治疗金黄色葡萄球菌感染提供用药及分子生物学依据。方法 收集2015年1月—2015年6月中山市7家医院分离到的金黄色葡萄球菌,使用ATB半自动细菌鉴定及药敏分析仪(法国梅里埃)对分离到的菌株进行鉴定及药敏试验,使用PCR技术对耐甲氧西林金黄色葡萄球菌(MRSA)的耐药基因进行检测。结果 7家医院共分离到89株金黄色葡萄球菌,其中MRSA检出33株,检出率为37.1%。金黄色葡萄球菌主要来源于呼吸内科(32株,36.0%)、骨科(20株,22.5%),主要分离自痰(41株,46.1%),伤口分泌物(16株,18%),对万古霉素、替考拉宁、奎奴普丁/达福普丁、复方新诺明、左氧氟沙星、诺氟沙星具有较高敏感性,MRSA对常用抗菌药物耐药率高于甲氧西林敏感金黄色葡萄球菌。共有32株MRSA检出bla_mecA基因,检出率为97%。结论 MRSA耐药情况较为严峻,临床科室应根据微生物培养报告合理使用抗菌药物。bla_mecA基因在MRSA检出较高,是MRSA主要的耐药机制。

Objective To analyze clinical distribution of Staphylococcus aureus infections from 7 hospitals in Zhongshan city, as well as to provide basis of empirical treatment and molecular biology for Staphylococcus aureus infections. Methods Staphylococcus aureus were collected from January 2015 to June 2015 in Zhongshan city, and then the strains were identified and tested antibiotic susceptibility by using ATB semiautomatic analyzer(Merieux). Resistance gene of methicillin-resistant Staphylococcus aureus(MRSA) was detected by polymerase chain reaction. Results 89 strains of Staphylococcus aureus were isolated from 7 hospitals and with prevalence of 33 strains of MRSA. Of all strains, 32(36.0%) were isolated from respiratory medicine and 20(22.5%) from orthopedics. 41(46.1%) strains of Staphylococcus aureus were isolated from sputum and 16(18.0%) from wound secretion. 89 strains of Staphylococcus aureus had highly susceptibility to vancomycin, teicoplanin, quinupristin/dalfopristin, cotrimoxazole, levofloxacin, norfloxacin. Resistance rates to commonly used antimicrobial drugs of MRSA were significantly higher than methicillin-sensitive. A total of 32 MRSA were detected carrying bla_mecA gene with the detection rate of 97%. Conclusion Clinical departments should be based on microbial culture report for rational use of antibiotics because of MRSA with more serious drug resistance. The gene of bla_mecA is the main mechanism of resistance for MRSA.