The integrons and insertion sequence common region 1 (ISCR1) of 329 carbapenem-non-susceptible Gram-negative bacilli, excluding 60 Stenotrophomonas maltophilia strains, in fecal specimens from 5000 patients in southern China were studied. A total of 205 (62.3%) class 1 integron-positive strains and 126 (61.5%) gene-cassette-positive strains were detected, and 44 types of gene-cassette arrays were found, including 8 types described for the first time in the corresponding species and one gene cassette originally detected in integrons, a putative helicase. A total of 9 types of 21 ISCR1-linked resistance gene regions in 35 ISCR1-positive bacilli and 2 previously unreported types of complex class 1 integrons were detected. All strains were extensively drug-resistant. Class 1 integrons and ISCR1 were widely disseminated and these structures carried various resistance genes, which were closely associated with mechanisms of multidrug resistance, including carbapenem resistance.

Objective Spinal cord injury-induced neuropathic pain(NP)does not respond well to any existing therapies clinically.This study aimed to explore the molecular metabolisms neuropathic pain by observing the expressions of the N -methyl-D-as-partic acid receptor 2B(NR2B),tumor necrosis factor-alpha(TNF-α)and brain-derived neurotrophic factor(BDNF)in different sta-ges of the condition in SD rats. Methods Thirty-six adult SD rats were randomly divided into a control,a sham operation and an NP model group.The NP model was established by spinal nerve ligation. At 5 and 14 days after modeling,the hindlimb motor function and paw withdrawal threshold(PWT)of the rats were evaluated by the open-field test and the expressions of NR 2B,TNF-αand BDNF in the dor-sal root ganglia of the L4-6 spinal cord were determined by Western blot. Results At 5 days after modeling,the open-field test showed a significantly shorter total distance of movement in the sham operation group than in the control([14 927.93 ±560.87]vs [18 225.15±371.76]mm,P<0.05)and even shorter in the NP model group([3 224.92±89.64]vs[18 225.15±371.76]mm, P<0.01).The time of activity was markedly decreased in the NP model group as compared with that in the control([203.48±19.94]vs [745.95±13.48]s,P<0.01),but with no statistically significant difference between the sham operation and control groups([727.93± 16.29]vs[745.95±13.48]s,P>0.05).At 14 days after modeling,both the total distance of movement and time of activity were re-markably shorter in the NP model than in the sham operation and control groups([3 395.53±96.12]vs[17 382.26±482.31]and [17 975.40±416.56]mm,P<0.01;[195.53±96.12]vs[739.31±18.36]and[775.20±16.84]s, P<0.01).The PWT showed no statistically significant difference among the three groups of rats before modeling(P>0.05)but markedly decreased in the NP model as compared with the sham operation and control groups(P<0.01),with no significant difference between the latter two(P>0.05).The expressions of NR2B,TNF-αand BDNF were remarkably up-regulated in the NP model group in comparison with the sham operation and control groups at 5 days after modeling(P>0.05)and even more significantly at 14 days(P<0.01). Conclusion NR2B and BDNF may be involved in the development and progression of neuropathic pain in rats.

目的 研究氯胺酮干预急性肺损伤炎症反应的最佳剂量和应用方法.方法 选择50只健康成年雄性Wistar大鼠,体质量(180±20)g,随机分为空白组(0.9％NaCl 2ml/h)、模型组(5mg/kg LPS)、小剂量氯胺酮组(5mg/kg LPS+ 5mg/kg氯胺酮)、中等剂量组(5mg/kg LPS+ 10mg/kg氯胺酮)和大剂量组(5mg/kg LPS+20mg/kg氯胺酮)各10只,检测血清肝肾功能、肺组织炎症因子及氧化因子水平.结果 中等剂量和大剂量组大鼠肺组织TNF-α、IL-10、IL-6、IL-8、MIP-2、iNOS、MDA、NO2-/NO3-水平及NF-κB结合活性较模型组及小剂量组显著降低(P＜0.05),中等剂量和大剂量组比较无差异(P＞0.05).中等剂量组大鼠血清ALT、UN和Cr水平低于大剂量组(P＜0.05);与小剂量组和模型组比较无差异(P＞0.05).结论 中等剂量及大剂量氯胺酮对ALI大鼠有强效抗炎和抗氧化应激作用,中等剂量的治疗安全性更具优势.

BACKGROUND:Integron, ISCR1 and complex class 1 integrons lead bacteria to become resistant to antibiotic regimens. The aim of this study was to detect common mobile genetic elements of multidrug-resistant Gram-negative bacilli and evaluate the genotyping of these bacilli in blood specimens from septicemia patients in southern China.METHODS:A total of 837 Gram-negative bacilli including 578 strains containing Enterobacteriaceae and 259 strains containing non-fermentative bacilli were investigated in blood samples collected from septicemia patients between 2011 and 2014 in southern China. Mobile genetic elements, such as class 1 integrons, the insertion sequence common region 1 (ISCR1), and complex class 1 integrons, were detected from the 837 strains.RESULTS:Twenty-seven types of gene cassette arrays were found among 837 strains in which 492 (58.8%) class 1 integron-positive isolates and 254 (51.6%) gene cassette-positive isolates were found, including the first description of two types, aacA4-bla IMP-1-bla OXA-30-catB3 and aac(6')-II-aadA13-cmlA8-bla OXA-10, in the corresponding species and two gene cassettes, putative helicase and aadA-like, originally detected in integrons. Twelve types of ISCR1-linked resistance gene regions in 196 ISCR1-positive bacilli and seven different types of complex class 1 integron-positive strains were obtained including four distinct complex class 1 integrons that have never been described in any species. Enterobacterial repetitive intergenic consensus (ERIC)-PCR fngerprinting showed that isolates with identical gene profles were clonally unrelated.CONCLUSION:Our results indicated that we should pay more attention to enhance the quality of infection control measures and prevent hospital infection, so as to avoid the outbreak of multidrug-resistant Gram-negative bacilli.

Objective So far, little is known about the mechanisms of molecular biology of primary frozen shoulder (PFS). This study was to investigate the effects of the single nucleotide polymorphisms ( SNP ) of matrix metalloproteinase-3 ( MMP-3) and col-lagen type Ⅰ alpha 1 ( COL1A1 ) on the susceptibility of PFS . Methods We recruited 42 PFS patients from the Department of Or-thopedics , Nanjing Jinling Hospital between January and July 2016 and 50 healthy controls.We genotyped the SNPs rs679620 and rs522616 of MMP-3 and rs1107946 of COL1A1 in the two groups of subjects. Results The genotype distribution of the MMP-3 SNP rs679620 showed no significant correlation with the susceptibility of PFS (P>0.05), and no statistically significant difference was found between allelic A and G (OR=0.54, 95%CI:0.30-0.99, P=0.044).The COL1A1 SNP rs1107946 and allelic frequencies were not correlated with the susceptibility of PFS , either ( P=0.994 and 0.920) .The recessive heredity model GG exhibited no statistically sig-nificant difference from GT, TT, and GT+TT, and nor allelic T from G (P>0.05). Conclusion Allelic A of the MMP-3 SNP rs679620 is a protective factor against the risk of PFS .Neither the MMP-3 SNP rs522616 nor COL1A1 rs1107946 is associated with the susceptibility of the disease .

目的 探讨载万古霉素硫酸钙复合骨形态发生蛋白2(BMP-2)治疗创伤性骨髓炎的临床效果.方法 回顾性分析2013年1月至2015年12月南京军区南京总医院收治的59例创伤性骨髓炎患者的临床资料,男27例,女32例;年龄21~62岁,平均年龄37.6岁;骨髓炎按Cierny-Mader分类:Ⅲ型37例、Ⅳ型22例;一期行内固定治疗39例、外固定支架固定20例.彻底清创后病灶内植入载万古霉素硫酸钙复合BMP-2,所有患者均改用外固定支架固定骨折断端,其中17例软组织缺损或骨缺损较大者采用带蒂转移皮瓣覆盖创面或肌筋膜瓣填塞髓腔.结果 所有患者均获随访,平均随访时间13.4个月(12~23个月).创面愈合良好,无窦道、软组织缺损及骨外露;骨折均获愈合,无感染复发,骨愈合时间5~14个月(平均7.8个月).3例患者因骨缺损范围较大而致骨折延迟愈合,二期植入自体髂骨后骨折愈合.末次随访时按Johner-Wruhs评分标准评定疗效:优54例、良5例,优良率100%;所有患者均可负重行走,无明显疼痛及畸形.结论 载万古霉素硫酸钙与BMP-2复合植入模式治疗创伤性骨髓炎,成骨可靠,安全性高,抗感染效果满意.

BACKGROUND:The connection structure of class 1 integron and insertion sequence common region 1 (ISCR1) is called "complex class 1 integrons" or "complex sul1-type integrons", which is also known to be associated with many resistance genes. This structure is a powerful gene-capturing tool kit that can mobilize antibiotic resistance genes. In order to look for and study the structure among clinical multidrug-resistant (MDR) Gram-negative isolates, 63 isolates simultaneously harbored class 1 integron and ISCR1-linked resistance genes were isolated from 2309 clinical non-redundant MDR Gram-negative isolates in Nanfang Hospital in 2008-2013. The connecting regions between the class 1 integrons and ISCR1 were examined using PCR and DNA sequencing to determine the structures in these isolates.RESULT:The two elements (the variable regions of the class 1 integron structures and the ISCR1-linked resistance genes) are connected in series among 63 isolates according to long-extension PCR and DNA sequencing. According to the kinds and permutations of resistance genes in the structure, 12 distinct types were identified, including 8 types that have never been described in any species. Several types of these structures are similar with the structures of other reports, but not entirely same.CONCLUSION:This study is the first to determine the structure simultaneously harboring class 1 integron and ISCR1-linked resistance genes by detecting the region connecting class 1 integrons and ISCR1 in a large number of MDR bacteria. These structures carrying various resistance genes were closely associated with multidrug resistance bacteria in Southern China.

Objective To observe the clinical effect of extract from rabbit skin inflamed by vaccinia virus (ERSIVV) in the limb salvage treatment of diabetic foot (DF).Methods From January 2013 to June 2015,46 DF patients with infection who needed limb salvage treatment in our hospital were selected and divided into the observation group and the control group according to treatment program,23 cases in each group.All of patients were graded according to the Wagner and accepted surgical treatment.Then observation group received the ERSIVV,the control group was given mecobalamin injection.The clinical efficacy and complications of the two groups were compared.Results The VAS scores of the two groups decreased after treatment,and the VAS scores 2 weeks and 3 weeks after treatment in the observation group were lower than those of the control group,the total effective rates of the pain improvement in the observation group were 82.61％ and 86.96％,which were higher than those in the control group (69.57％ and 73.91％) (P ＜ 0.05).The scores of neurological symptoms and signs of the two groups decreased after treatment,and the scores in the observation group were lower than those in the control group,the difference was statistically significant (P ＜ 0.05).After treatment,the nerve conduction velocity in the two groups all increased,and the change of the observation group was more obvious (P ＜ 0.05).There were no obvious complication and adverse reaction in the two groups during the treatment period.Conclusion The ERSIVV can effectively improve the pain,neurologic symptoms and signs of the patients with DF infection after the limb salvage treatment,promote the recovery of nerve conduction speed,it has good clinical efficacy and safety.

Objective To detect and have homology analysis of resistant genes in clinical isolates of resistant Acinetobacter Bauman (CRAB).Methods Collected 40 clinical isolates of CRAB in Zhongshan Hospital of Traditional Chinese Medicine from July 2012 to December 2013,using PCR method for the amplification of the gene of carbon,sequence analysis.Applica-tion of ERIC-PCR to study the transmission mechanism.Results 40 strains of crab all isolates and detected containing oxa-51-like gene for 40 strains (100%),the gene oxa-23-like geneand for 38 strains (95%),were not detected in NDM,VIM, IPM,gene OXA-24-like and OXA-58-like.ERIC-PCR clustering analysis of similarity was greater than 0.8,that was the same colonies.The 40 strains of bacteria at a similar level were clustered into 33 taxa (clone).Conclusion The clinical iso-lates of CRAB mainly carry on the effective method of OXA-23-like,OXA-51-like,and ERIC-PCR for the homology analysis of Acinetobacter Bauman.

Objective To knock down hemolysin hlyA gene and insert green fluorescence protein gene of vibrio cholerae vaccine candidate NFYY101. Methods Amplified fragments of hlyA gene upstream (hlyAup) and downstream (hlyAdown),lacz-GFPuv,and hlyAup-laczGFPuv-hlyAdown, and plasmids treated with specific enzymes were utilized to construct recombinant plasmids pUC18-hlyAup-laczGFPuv-hlyAdown and pCVD442-hlyAup-laczGFPuv-hlyAdown. Following the construction of the recombinant suicide plasmids ,a vaccine candidate was constructed by homologous recombination ,while SM10λpir carrying pCVD442- hlyAup-laczGFPuv-hlyAdown was utilized as the donor strain to transfect NFYY101. Results Construction of recombinant suicide plasmids pCVD442- hlyAup-laczGFPuv-hlyAdown was satisfactory that hemolysin hlyA gene was knocked out and green fluorescence protein gene was successfully inserted of vibrio cholerae vaccine candidate NFYY101. Conclusion Construction of the recombinant suicide plasmid pCVD442-hlyAup-laczGFPuv-hlyAdown successfully can be used for knocked out the hlyA gene and added green fluorescence protein gene as genetic marker into the chromosomal DNA of vibrio cholerae vaccine candidate.

Objective To investigate the drug resistance of carbapenem‐resistant Acinetobacter baumannii(CRAB) clinically iso‐lated from hospital ,OXA carbapenemases gene and epidemiological characteristics .Methods A total of 40 clinically isolated strains of CRAB were collected in this hospital from July 2011 to December 2012 .oxa‐23 ,oxa‐24 ,oxa‐51 and oxa‐58 were amplified and se‐quenced by adopting the PCR method .Results Forty strains of CRAB had the highest sensitivity to polymyxin B and minocycline among 16 kinds of drugs ,with the resistance rate less than 5 .0% ;29 strains(72 .5% ) were positive in the modified Hodge test ;the oxa‐51 gene was detected in all strains ,accounting for 100 .0% ,in which 38 strains(95 .0% ) contained oxa‐23 gene ,no oxa‐24 and oxa‐58 genes were detected .Conclusion oxa‐51 and oxa‐23 genes are the main genotypes of epidemic CRAB in Zhongshan Munici‐pal Hospital of Traditional Chinese Medicine .

目的:阐明耐碳青霉烯鲍曼不动杆菌(CRAB)主要耐药机制和分子流行病学特征.方法:PCR方法检测常见碳青霉烯酶基因,PCR-RELP分型及基因测序检测整合子Ⅰ和ISCR1,ERIC-PCR及聚类分析研究克隆相关性.结果:碳青霉烯酶基因NDM、IMP、VIM、OXA-23-like、OXA-24-like、OXA-5 1-like、OXA-58-like检出率分别为4.1％、22.9％、26.0％、67.7％、63.5％、46.8％、65.6％.整合子Ⅰ可变区分为12个耐药基因盒排列类型,A6和A12首次在鲍曼不动杆菌中发现;ISCR1可变区分为6个类型.将耐药机制、ERIC-PCR及聚类分析结合临床资料综合分析,CRAB感染患者均为散发,粪便中CRAB不是临床感染来源.结论:CRAB主要耐药机制是产碳青霉烯酶及整合子Ⅰ、ISCR1携带率高,ERIC-PCR基因分型结合耐药机制研究适用于分子流行病学调查.