The clinical strain of Pseudomonas aeruginosa XM8 harbored multiple RND-type antibiotic efflux pump genes and a novel integron In4881 on its plasmid pXM8-2, rendering it resistant to nearly all conventional antibiotics except colistin. The resistance was primarily attributed to the inactivation of the oprD gene and overexpression of several efflux pump genes, including mexAB-oprM, mexCD-oprJ, oprN-mexFE, and mexXY. In this study, the XM8 strain was comprehensively characterized using various methods. Antimicrobial susceptibility testing was performed using the BioMerieux VITEK2 system and manual double dilution methods. Gene expression levels of efflux pump-related genes were analyzed via quantitative real-time PCR. The bacterial chromosome and plasmid were sequenced using both Illumina and Nanopore platforms, and bioinformatics tools were employed to analyze mobile genetic elements associated with antibiotic resistance. The pXM8-2 plasmid containsed multiple mobile genetic elements, including integrons (In4881, In334, In413) and transposons (Tn3, TnAs1, TnAs3). Notably, In4881 was reported for the first time in this study. The presence of these elements highlights the potential for horizontal gene transfer and further spread of antibiotic resistance. Given the strong resistance profile of the XM8 strain, effective measures should be implemented to prevent the dissemination and prevalence of such multidrug-resistant bacteria.

Purpose:This study aimed to investigate the pathogenic characteristics of Vibrio parahaemolyticus in Taizhou City, China, to provide a scientific basis for the diagnosis and control of Vibrio parahaemolyticus in the region. Methods:Bacterial strains were isolated from stool specimens of diarrheal patients at the First People's Hospital of Wenling City. The virulence genes, serotypes, and antibiotic susceptibility of these strains were determined. Results:From June 1, 2020 to May 31, 2023, a total of 97 strains of Vibrio parahaemolyticus were isolated from stool specimens of diarrheal patients at the First People's Hospital of Wenling City. The results of virulence gene detection showed that 87.6% of the strains were thermostable direct hemolysin (TDH; tdh)+, thermolabile hemolysin (tlh)+, and tdh-related hemolysin (trh)-, 11.3% were tdh - tlh + trh -, and 1.0% were tdh + tlh + trh +. Six different serotypes were identified, with O3:K4 and O4:K4 being the most common, accounting for 42.3% and 32.0%, respectively. The other serotypes identified were O3:K6, O4:K8, and O1:KUT, accounting for 11.3%, 10.3%, and 3.1%, respectively. Furthermore, the results of antimicrobial susceptibility tests showed that most strains were sensitive to the tested drugs, while some strains exhibited resistance to cefuroxime (32.6%) and cefazolin (43.6%). Conclusion:The majority of the isolated Vibrio parahaemolyticus strains carried the tdh gene, and the O3:K4 serotype dominated in Taizhou City from 2020 to 2023, followed by the O4:K4 serotype. Furthermore, some strains exhibited resistance to cefuroxime and cefazolin.

Methicillin-resistant Staphylococcus aureus (MRSA) is a common pathogen contributing to healthcare-associated infections, which can result in multiple sites infections. The epidemiological characteristics of MRSA exhibit variability among distinct regions and healthcare facilities. The aim of this study was to investigate the molecular epidemiology and nosocomial outbreak characteristics of MRSA in a county-level hospital in China. A total of 130 non-repetitive MRSA strains were collected from December 2020 to November 2021. Whole-genome sequencing (WGS) was performed to identify antimicrobial resistance and virulence factors. Phylogenetic analysis was conducted to ascertain genetic diversity and phylogenetic relationships. Independent transmission scenarios were determined by the phylogeny derived from single nucleotide polymorphisms (SNPs) within the core genome. All the MRSA isolates were collected from the intensive care unit (30.00%, 39/130), the department of otorhinolaryngology (10.00%, 13/130) and the department of burn unit (9.23%, 12/130). The clinical samples mainly included phlegm (53.85%, 70/130), purulent fluid (24.62%, 32/130), and secretions (8.46%, 11/130). The resistance rates to erythromycin, clindamycin and ciprofloxacin were 75.38, 40.00, and 39.23%, respectively. All the isolates belonged to 11 clonal complexes (CCs), with the major prevalent types were CC5, CC59, and CC398, accounting for 30.00% (39/130), 29.23% (38/130), and 16.92% (22/130), respectively. Twenty sequence types (STs) were identified, and ST59 (25.38%, 33/130) was the dominant lineage, followed by ST5 (23.84%, 31/130) and ST398 (16.92%, 22/130). Three different SCCmec types were investigated, most of isolates were type IV (33.85%, 44/130), followed by type II (27.69%, 36/130) and type III (0.77%, 1/130). The common clonal structures included CC5-ST5-t2460-SCCmec IIa, CC59-ST59-t437-SCCmec IV and CC398-ST398-t034-SCCmec (−), with rates of 16.92% (22/130), 14.62% (19/130), and 13.84% (18/130), respectively. Only 12 panton-valentine leucocidin (PVL) positive strains were identified. Two independent clonal outbreaks were detected, one consisting of 22 PVL-negative strains belongs to CC5-ST5-t2460-SCCmec IIa and the other consisting of 8 PVL-negative strains belongs to CC5-ST5-t311-SCCmec IIa. Overall, our study indicated that the CC5 lineage emerged as the predominant epidemic clone of MRSA, responsible for nosocomial outbreaks and transmission within a county-level hospital in China, highlighting the necessity to strengthen infection control measures for MRSA in such healthcare facilities.

ObjectiveThe emergence of clinical Klebsiella pneumoniae strains harboring acrAB-tolC genes in the chromosome, along with the presence of two repetitive tandem core structures for blaKPC-2 and blaCTX-M-65 genes on a plasmid, has presented a significant clinical challenge.MethodsIn order to study the detailed genetic features of K. pneumoniae strain SC35, both the bacterial chromosome and plasmids were sequenced using Illumina and nanopore platforms. Furthermore, bioinformatics methods were employed to analyze the mobile genetic elements associated with antibiotic resistance genes.ResultsK. pneumoniae strain SC35 was found to possess a class A beta-lactamase and demonstrated resistance to all tested antibiotics. This resistance was attributed to the presence of efflux pump genes, specifically acrAB-tolC, on the SC35 chromosome. Additionally, the SC35 plasmid p1 carried the two repetitive tandem core structures for blaKPC-2 and blaCTX-M-65, as well as blaTEM-1 with rmtB, which shared overlapping structures with mobile genetic elements as In413, Tn3, and TnAs3. Through plasmid transfer assays, it was determined that the SC35 plasmid p1 could be successfully transferred with an average conjugation frequency of 6.85 × 10-4.ConclusionThe structure of the SC35 plasmid p1 appears to have evolved in correlation with other plasmids such as pKPC2_130119, pDD01754–2, and F4_plasmid pA. The infectious strain SC35 exhibits no susceptibility to tested antibioticst, thus effective measures should be taken to prevent the spread and epidemic of this strain.

Background: Numerous studies have recorded the function of long noncoding RNAs (lncRNAs) in cancer development, including lung adenocarcinoma (LUAD). Previous studies have reported the crucial role of lncRNA PTPRG antisense RNA 1 (PTPRG-AS1) in various cancers. However, the role of PTPRG-AS1 in LUAD remains unknown. Materials and Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) was applied for detecting PTPRG-AS1 expression in LUAD cell lines. Functional assays and in vivo experiments explored cell proliferation, whereas flow cytometry analysis was used to detect cell cycle. In addition, fluorescent in situ hybridization (FISH) and subcellular fractionation assay measured the localization of PTPRG-AS1 in LUAD cells. RNA pulldown, luciferase reporter, and RNA immunoprecipitation (RIP) assays were used to investigate the interaction of PTPRG-AS1/miR-124-3p/cyclin D1 (CCND1) axis. Results: PTPRG-AS1 expression was notably high in LUAD cell lines. PTPRG-AS1 knockdown suppressed cell proliferation and cycle as well as the level of CCND1. Moreover, miR-124-3p was the mutual target of PTPRG-AS1 and CCND1. In addition, PTPRG-AS1 sponged miR-124-3p to upregulate CCND1 in LUAD cells. Moreover, miR-124-3p depletion reversed the suppression of PTPRG-AS1 silence on LUAD cell behaviors, but then CCND1 knockdown countervailed the promoting influence of downregulated miR-124-3p. Conclusions: PTPRG-AS1 propels cell proliferation and cell cycle of LUAD by targeting miR-124-3p/CCND1 axis.

目的 分析浙江省温岭市2010至2020年副溶血弧菌大流行株感染性腹泻的流行病学状况.方法 选取2010-2020年浙江省温岭市食源性疾病检测哨点医院就诊,并报告至国家食源性疾病监测系统的急性腹泻患者900例,根据肠道细菌培养结果统计副溶血弧菌大流行株感染的检出率,分析其危险因素及主要血清型构成分布情况.结果 900例急性腹泻患者中有500例检测出副溶血弧菌大流行株,检出率为55.56％,其血清型主要为O3:K6(占45.45％～85.11％).单因素分析显示:副溶血弧菌大流行株感染性急性腹泻与性别、职业类型、家庭月收入无关(P>0.05),而与年龄、食品暴露史、厨房抹布平均更换频率、单独餐具、洗手方式、是否使用家用带盖垃圾桶、是否为公共厨房、患病季节有关(P<0.05).多因素Logistics回归分析显示:年龄、食品暴露史、厨房抹布平均更换频率、单独餐具、洗手方式、是否使用家用带盖垃圾桶、是否为公共厨房、患病季节为影响副溶血弧菌大流行株感染性急性腹泻的主要危险因素(P<0.05).结论 副溶血弧菌大流行株感染是急性腹泻的主要病原微生物,其主要血清型为O3:K6,其发生与卫生习惯、卫生环境有关.可根据这些因素采用相应的预防措施,以减少副溶血弧菌大流行株感染性急性腹泻的发生.

目的:利用生物信息学方法鉴定参与肝细胞癌(HCC)发生发展的候选基因及其潜在机制.方法:从基因表达数据库(GEO)中获得GSE19665、GSE41804和GSE84402,通过GEO2R工具和Venn图软件选择HCC组织与正常肝组织之间差异表达基因(DEG).对这些DEG进行京都基因与基因组百科全书(KEGG)通路、基因本体(GO)富集分析、蛋白-蛋白相互作用(PPI)、生存分析(Kaplan-Meier)和基因表达谱交互式分析(GEPIA)验证.结果:3个数据集中共有139个一致表达的DEG,包括36个上调基因和103个下调基因.28个核心基因中有26个的预后明显较差,且有22个在HCC组织中高表达,4个基因(CCNB1、CDK1、BUB1和PT-TG1)显著富集于细胞周期途径.结论:HCC预后不良的4个显著上调的DEG可能是HCC患者的潜在治疗靶标.

目的 优化高橙皮中柠檬苦素的超声提取工艺.方法 在单因素试验的基础上,以声强、乙醇体积分数、液料比、提取时间为影响因素,柠檬苦素提取率为评价指标,正交试验优化提取工艺.结果 最佳条件为声强0.255 6 W/cm2,乙醇体积分数70％,料液比30∶1,提取时间30 min,柠檬苦素提取率为60.90 mg/100 g.结论 该方法合理可行,可用于超声提取高橙皮中柠檬苦素.

Background: Streptococcus pneumoniae is a leading pathogen worldwide and its pathogenesis remains unclear. Objectives: The study aimed to investigate the secretion of interleukin-8 (IL-8) and soluble intercellular adhesion molecule-1(ICAM-1) from A549 pneumocytes stimulated by different S. pneumoniae strains and the mechanism of blood-derived S. pneumoniae (bd-SP) in invading the blood system. Methods: Twenty-three clinical strains of S. pneumoniae isolated in 2009 along with ATCC49619 were cultured and suspended in 2018. Then, A549 pneumocytes were resuspended after culture and collection, and inoculated into culture plates. Streptococcus pneumoniae suspensions were then inoculated. Normal saline was blank control. The plates were incubated for four and eight hours. Then, the suspensions were collected and centrifuged. The supernatant was analyzed for IL-8 and ICAM-1 by ELISA. Results: The concentrations of IL-8 were 180.6, 188.5, 223.4 +/- 19.9, and 230.3 +/- 38.6 pg/mL for blank control,ATCC49619, blood-derived S. pneumoniae (bd-SP) and sputum-derived S. pneumoniae (sd-SP) at four-hour stimulation, respectively, and 249.2, 275.7, 224.0 +/- 27.8, and 242.3 +/- 33.1 pg/mL at eight-hour stimulation. The concentrations of ICAM-1 were 14.8,12.1,19.9 +/- 17.2, and 26.1 +/- 28.6 ng/mL for blank control, ATCC49619, bd-SP, and sd-SP at 4 hours, respectively, and 32.6, 150.8, 69.4 +/- 45.1, and 58.7 +/- 30.1 ng/mL at 8 hours. Conclusions: Streptococcus pneumoniae could upregulate the secretion of IL-8 and ICAM-1 from A549 pneumocytes. No significant difference was found between bd-SP and sd-SP which was in contrast to what was found between clinical S. pneumoniae and ATCC49619. Host response may not be a vital factor for different S. pneumoniae infections.

In this work, we aim to develop and validate a fast, simple, and sensitive method for the quantitative determination of flibanserin and the exploration of its pharmacokinetics. Ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was the method of choice for this investigation and carbamazepine was selected as an internal standard (IS). The plasma samples were processed by one-step protein precipitation using acetonitrile. The highly selective chromatographic separation of flibanserin and carbamazepine (IS) was realised using an Agilent RRHD Eclipse Plus C18 (2.1 × 50 mm, 1.8 µ) column with a gradient mobile phase consisting of 0.1% formic acid in water and acetonitrile. The analytes were detected using positive-ion electrospray ionization mass spectrometry via multiple reaction monitoring (MRM). The target fragment ions were m/z 391.3 → 161.3 for flibanserin and m/z 237.1 → 194 for carbamazepine (IS). The method was validated by linear calibration plots over the range of 100–120,000 ng/mL for flibanserin (R2 = 0.999) in rat plasma. The extraction recovery of flibanserin was in the range of 91.5–95.8%. The determined inter- and intra-day precision was below 12.0%, and the accuracy was from − 6.6 to 12.0%. No obvious matrix effect and astaticism was observed for flibanserin. The target analytes were long-lasting and stable in rat plasma for 12 h at room temperature, 48 h at 4 °C, 30 days at − 20 °C, as well as after three freeze–thaw cycles (from − 20 °C to room temperature). The proposed method has been fully validated and successfully applied to the pharmacokinetic study of flibanserin.

目的 分析不同细菌类型血流感染患者和抗菌治疗前后血清降钙素原(PCT)变化的差异,为PCT在临床细菌感染的初期用药及预后提供可靠的临床观察数据.方法 采用回顾性分析方法比较2015年1月-2016军12月163例血培养阳性患者抗菌治疗前及治疗24 h、48 h、72 h后的PCT水平.根据细菌类型分为革兰阳性菌组和革兰阴性菌组,比较抗菌治疗前PCT的差异,以及治疗前后PCT变化差异.结果 革兰阴性菌组的血清PCT水平明显高于革兰阳性菌组,差异有统计学意义(P＜0.01);不同细菌类型抗菌治疗24h、48 h、72 h后血清PCT变化率之间差异均有统计学意义(P＜0.05).结论 不同细菌类型血流感染患者具有不同的PCT变化规律,比较PCT值可作为血培养阳性预测因子,可指导临床早期感染的经验用药.不同类型细菌血流感染患者抗菌治疗后血清PCT下降程度不同,结合抗菌治疗前后PCT变化率可以预测患者预后.

目的 探讨[Gly14]-Humanin(HNG)预处理对局灶性脑缺血再灌注损伤后大鼠自由基代谢的影响,分析相关的神经功能保护机制.方法 将64只健康雄性SD大鼠随机分为HNG组、生理盐水组、模型组及假手术组,每组16只.采用改良的Zea-longa线栓法建立大脑中动脉缺血再灌注损伤模型,生理盐水组与假手术组大鼠术前3d给予3μL生理盐水连续3d尾静脉注射,每日1次;HNG组给予100 nmol/L HNG 5 μL,而模型组除正常饲养外术前不接受任何处理.各组大鼠在缺血2h再灌注24 h后检测脑组织谷胱甘肽(GSH)及丙二醛(MDA)的含量水平、超氧化物歧化酶(SOD)的活性,原位末端标记染色观察凋亡细胞数并进行统计学分析.结果 与假手术组相比,其余三组大鼠的GSH含量水平及SOD的活性均降低,而MDA含量水平及细胞凋亡率升高,差异均有显著性(P＜0.01～0.05);与生理盐水组及模型组比较,MDA含量水平及细胞凋亡率均降低,而HNG组大鼠的GSH含量水平及SOD的活性均升高,差异均有显著性(P＜0.01～0.05).结论 HNG预处理减轻脑缺血再灌注损伤过程中的增加脑组织抗氧化物质SOD的活性及GSH含量水平,减少神经细胞的凋亡.

OBJECTIVE To understand the distribution and drug resistance of pathogens causing the urinary tract infections in the patients of neurology department so as to guide the reasonable clinical use of antibiotics.METHODS A total of 754specimens submitted from Oct 2009to Oct 2011were analyzed,VITEK-2Compact automatic micro-analyzer system was used to perform the identification and drug susceptibility testing,and the drug resistance of the pathogens was analyzed.RESULTS A total of 310strains of pathogens were isolated,excluding the repeat specimens from the same patient,the main pathogens were Escherichia coli,Klebsiella pneumoniae,and Enterococcus,accounting for 33.9%,9.0%,and 13.8%,among which the ESBLs-producing E.coli accounted for 39.2%,ESBLs-producing K.pneumoniae 41.3%.The drug susceptibility rates of the E.coli and K.pneumoniaeto imipenem were 100.0%,the drug susceptibility rates to cefoperazone/sulbactam were more than 95.0%;there was one strain of imipenem-resistant Enterobacter cloacae,with the modified Hodge test positive;Staphylococcus aureus and other gram-positive bacteria were 100.0%sensitive to vancomycin,the detection rate of the methicillin-resistant Staphylococcus aureus(MRSA)was 38.5%;the detection rate of the fungi was 11.3%.CONCLUSION The opportunistic pathogens remain the common pathogens causing the urinary tract infections in the neurology department and have produced certain drug resistance to the commonly used antibiotics.It is necessary to strengthen the monitoring of the drug resistance,standardize the operation,and guide the clinical medication on the basis of the drug susceptibility testing so as to effectively control the nosocomial infections.

目的 通过酶联免疫斑点法(ELISPOT)检测,探讨甲型H1N1流感患者Th1/Th2细胞的变化及其临床意义.方法 对40例H1N1患者、42例流感样患者及45例健康体检者采用ELISPOT技术检测Th1/Th2细胞,同时进行外周血中的白细胞、淋巴细胞及其亚群计数.结果 H1N1患者的白细胞、淋巴细胞、T细胞计数(包括T4、T8)与流感样患者及健康体检者比较明显偏低(P＜0.05);B细胞计数与健康体检者比较明显偏低(P＜0.05);而H1N1患者NK细胞与流感样患者、健康体检者比较,差异均无统计学意义(均P ＞0.05).H1N1组、流感样组和健康对照组中IFN-γ特异的ELISPOT检测SFC数量M值分别为23、36.5、43,H1N1组与流感样组、健康对照组比较,差异均有统计学意义(均P＜0.05).H1N1组、流感样组和健康组中IL-4特异的ELISPOT检测SFC数量M值分别为36.5、27、23,H1N1组与健康对照组比较,差异有统计学意义(P＜0.05);但与流感样组比较,差异无统计学意义(P ＞0.05).结论 H1N1感染可引起机体淋巴细胞及其亚群的数量发生异常改变,引起患者机体免疫系统的紊乱.通过ELISPOT技术可从单细胞水平明确CD4+T淋巴细胞亚群中的Th1和Th2细胞的异常改变.

<正>氨基糖苷类抗生素通过作用于细菌核糖体30S亚基的16S rRNA高度保守的A位点,干扰细菌蛋白质的合成,从而引起细菌死亡。氨基糖苷类抗生素具有杀菌作用,抗菌谱广,较长抗生素后效应等特点,而且还与其他抗菌药物有协同作

目的研究帕瑞昔布钠对子宫全切术后镇痛效果和应激反应的影响。方法选择在全身麻醉下行子宫切除术患者100例,分为帕瑞昔布钠组和对照组,各50例。两组术后均采用芬太尼行自控静脉镇痛,帕瑞昔布钠组于术毕即刻,术后12、24及36h分别静脉注射帕瑞昔布钠40mg;对照组上述时点静脉注射0.9%氯化钠注射液3ml。于麻醉前(T0),术后2h(T1)、12h(T2)、24h(T3)及48h(T4)采用放射免疫分析法测定两组血浆肾素、血管紧张素Ⅱ、醛固酮及皮质醇水平,于T1～T4点记录两组视觉模拟镇痛评分(VAS)。结果帕瑞昔布钠组在T2、T3及T4时点VAS均明显低于对照组(P<0.05);血浆肾素、血管紧张素Ⅱ、醛固酮及皮质醇浓度,帕瑞昔布钠组T0时点与T2、T3及T4时点差异均无统计学意义(均P>0.05);对照组T2及T3比T0明显增高(P<0.05);T2及T3时点帕瑞昔布钠组与对照组差异均有统计学意义(均P<0.05)。结论帕瑞昔布钠术后镇痛效果确切,可明显减轻手术创伤激活的过度应激反应,对子宫全切术患者的康复具有重要意义。

目的:比较3种不同梅毒血清学实验方法在临床中的应用价值。方法:应用甲苯胺红不加热血清试验(TRUST)、梅毒螺旋体明胶凝集试验(TPPA)、梅毒酶联免疫吸附试验(ELISA)检测血清标本中的梅毒螺旋体抗体。结果:TRUST、TPPA、ELISA的阳性率分别为86.67%、98.33%、98.89%。特异性分别为78.86%、96.57%、90.86%。同时发现ELISA检测时存在假阳性。结论:TRUST不适合作为梅毒的诊断。ELISA可作为输血和手术前检测的一种理想筛查方法。但对ELISA阳性标本应做TPPA检测,对ELISA检测阴性的标本则不需要TPPA确诊。