Phenotypic and Genotypic Characterization of Extended Spectrum β-Lactamase Klebsiella pneumoniae and Fluorescent Pseudomonas spp. Strains from Market Garden Products and Their Watering Water in Benin (West Africa)

Market garden products can carry several types of microorganisms, and their consumption is the source of many cases of food poisoning. This work aimed to improve food safety in Benin. In characterizing strains of K. pneumoniae and fluorescent Pseudomonas spp. at the biochemical and molecular level, the target was to identify contaminated watering water and garden products sold during Cotonou in both the dry and rainy seasons. A total of 164 samples of market garden products and 22 samples of watering water were investigated. The results showed that 5.91% of market garden products and watering water were contaminated by K. pneumoniae and 20.43% by fluorescent Pseudomonas spp. During the dry season, cabbage was most contaminated by fluorescent Pseudomonas spp. (50%). Pool water was more contaminated with K. pneumoniae (17%). All isolated strains were resistant to both amoxicillin and penicillin. All strains of K. pneumoniae and fluorescent Pseudomonas spp. were not resistant to imipenem, and 22% of them produced penicillinase. Among the 49 strains producing penicillinase isolated, 64.29% and 21.43% carried blaTEM and blaSHV respectively while 14.28% carried blaCTX-M genes. In light of the previously-developed results and considering the importance of horticultural products in Beninese food habits, we must improve national awareness of the risk for foodborne illness.

External Bacterial Flora and Antimicrobial Susceptibility Patterns of Staphylococcus spp. and Pseudomonas spp. Isolated from Two Household Cockroaches, Blattella germanica and Blatta orientalis

A study was performed to estimate the prevalence of the external bacterial flora of two domestic cockroaches （Blattella germanica and Blatta orientalis） collected from households in Tebessa （northeast AIgeria）.Three major bacterial groups were cultured （total aerobic, enterobacteria, and staphylococci） from 14 specimens of cockroaches, and antibiotic susceptibility was tested for both Staphylococcus and Pseudomonas isolates. Culturing showed that the total bacterial load of cockroaches from different households were comparable （P〈0.001） and enterobacteria were the predominant colonizers of the insect surface, with a bacterial load of （2.1×10^5 CFU/insect）, whereas the staphylococci group was the minority. Twenty-eight bacterial species were isolated, and susceptibility patterns showed that most of the staphylococci isolates were highly susceptible to chloramphenicol, gentamycin, pristinamycin, ofloxacin, clindamycin, and vancomycin; however, Pseudomonas strains exhibited resistance to amoxicillin/clavulanic acid, imipenem, and the second-generation antibiotic cephalosporin cefuroxime.

基于改进YOLOv3-SPP算法的道路车辆检测

针对在城市道路场景下视觉检测车辆时,车辆密集和远处车辆呈现小尺度,导致出现检测精度低或者漏检的问题,提出了一种基于改进的YOLOv3-SPP算法,对激活函数进行优化,以DIOU-NMS Loss作为边界框损失函数,增强网络的表达能力。为提高所提算法对小目标和遮挡目标的特征提取能力,引入空洞卷积模块,增大目标的感受野。实验结果表明,所提算法在检测车辆目标时m AP提高了1.79%,也有效减少了在检测紧密车辆目标时出现的漏检现象。

FOSB、SPP1基因表达对经肝动脉介入化疗栓塞术治疗的中晚期肝癌生存期预测价值

目的探讨骨肉瘤原癌基凶同源物B(FOSB)、分泌性磷蛋白1(SPP 1)基因表达对经肝动脉介入化疗栓塞术治疗的中晚期肝癌患者术后生存期的预测价值。方法回顾性分析2016年1月—2017年10月在西安医学院第一附属医院进行诊治的中晚期肝癌患者115例作为研究对象,根据FOSB、SPP 1基因表达水平分为FOSB高表达组(n=52)、FOSB低表达组(n=63)及SPP 1高表达组(n=89)、SPP 1低表达组(n=26)。同时选取115例健康体检者为健康对照组。分析FOSB、SPP 1表达与中晚期肝癌患者临床病理特征的关系。对纳入研究的患者进行为期60个月的随访,Logistics风险回归模型分析影响患者生存期的危险因素。Kaplan-Meier生存曲线分析FOSB、SPP 1表达水平与患者生存预后的关系。结果肝癌患者外周血单个核细胞中FOSB mRNA表达水平明显低于健康对照组,SPP 1 mRNA表达水平明显高于健康对照组,差异具有统计学意义(P<0.05)。FOSB、SPP 1表达在中晚期肝癌患者肿瘤大小、Child-Pugh分级、淋巴转移、BCLC分期方面比较差异具有统计学意义(P<0.05)。单因素分析结果显示,存活组和死亡组在肿瘤大小、Child-Pugh分级、淋巴转移、BCLC分期、FOSB、SPP 1表达方面比较,差异均具有统计学意义(P<0.05)。肿瘤大小、Child-Pugh分级、淋巴转移、BCLC分期、FOSB低表达、SPP 1高表达均为中晚期肝癌患者生存期的独立危险因素(P<0.05)。随访时间60个月,患者生存率为17.39%(20/115),FOSB高表达组中位生存时间为39.7个月,明显高于FOSB低表达组的19.3个月(P<0.05);SPP 1低表达组中位生存时间为40个月,明显高于SPP 1高表达组的18个月(P<0.05)。结论FOSB在中晚期肝癌患者中表达明显下调,SPP 1表达上调,其对预测中晚期肝癌患者肝动脉介入化疗栓塞术治疗生存期具有一定的价值。FOSB、SPP 1有望成为评估介入术治疗患者预后的潜在指标,可协同肿瘤大小、Child-Pugh分级、淋巴转移、BCLC分期等临床指标预测或评估肝癌患者术后生存情况。

SPP1和PD-L1在子宫内膜癌组织中的表达及其预后价值

目的:分析分泌型磷蛋白1(secreted phosphoprotein 1,SPP1)和程序性死亡蛋白配体1(programmed cell death-ligand 1,PD-L1)在子宫内膜癌(endometrial carcinoma,EC)组织中的表达水平及与预后的关系。方法:通过免疫组织化学方法分析SPP1和PD-L1在84例EC组织和61例正常子宫内膜组织中的表达情况,分析SPP1和PD-L1表达与临床病理特征及预后的关系,利用UALCAN分析工具(https://ualcan.path.uab.edu/)分析癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中EC组织SPP1和PD-L1 mRNA的表达及与预后的关系。结果:EC组织SPP1阳性表达率高于正常子宫内膜组织(71.43%vs 36.07%,P=0.001),PD-L1阳性表达率低于正常子宫内膜组织(25.00%vs 42.62%,P=0.025);EC组织中,SPP1阳性表达率与组织学分级相关(P<0.05),PD-L1阳性表达率与淋巴结转移相关(P<0.05),SPP1与PD-L1表达呈正相关(列联系数r=0.237,P=0.026);EC组织中,不同SPP1和PD-L1表达患者总生存期无差异(均P>0.05)。TCGA数据库中生物信息学分析显示EC中SPP1 mRNA表达水平显著高于正常子宫内膜组织(P<0.01),PD-L1 mRNA表达水平与正常子宫内膜组织比较,差异无统计学意义(P=0.057);EC组织SPP1 mRNA和PD-L1 mRNA表达水平呈正相关(r=0.350,P<0.05);不同SPP1和PD-L1表达水平患者总生存期无差异(均P>0.05)。结论:SPP1在EC组织中的表达与PD-L1呈正相关,可能共同参与EC免疫微环境调控,SPP1和PD-L1均不是EC独立预后因子。

磷改性SPP沸石催化甲醇制丙烯反应

为开发甲醇制丙烯(MTP)反应的高性能催化剂,制备了不同硅铝比的SPP沸石,再进行磷改性处理。采用X射线衍射(XRD)、扫描电镜(SEM)、电感耦合等离子发射光谱(ICP)、氮气物理吸附、NH_(3)程序升温脱附(NH_(3)-TPD)和热重分析等手段表征了样品的孔结构和酸性质等,并比较了各催化剂在甲醇制丙烯(MTP)反应中的催化性能。结果表明:相比于普通ZSM-5沸石,SPP沸石具有更大的比表面积和孔容,可以显著提高分子的扩散速率和活性位点的可接触性,提高了反应性能。得益于弱碱性磷酸铵的孔道清洁作用,磷改性后的SPP沸石的总孔容均增加。同时,随着磷改性浓度的增加,SPP沸石的强酸量逐渐降低。磷改性得到的SPP沸石(磷的质量分数为0.2%)催化MTP反应时,丙烯选择性达到50.4%,C_(2)^(=)-C_(4)^(=)选择性达到72.1%,积炭量为1.4%。

栖稻假单胞菌(Pseudomonas oryzihabitans)研究进展

栖稻假单胞菌(P.oryzihabitans)是一类非发酵、需氧型革兰氏阴性杆菌,自1984年首次发现以来,在美国国家生物信息中心(NCBI)登记的菌株超过1500种。这些菌株广泛存在于患病的人类和动植物体内以及土壤、水等环境中。P.oryzihabitans的一些菌株作为人类和动植物共同的致病菌,对人类健康和农业发展造成潜在威胁;同时,某些菌株可作为植物根际促生菌种,推动农业生产绿色可持续发展。对栖稻假单胞菌的生物学特性、致病性及应用等进行综述,旨在为栖稻假单胞菌的防治和应用提供参考。

Characterization of 1-Aminocyclopropane-1-Carboxylate(ACC) Deaminase-Containing Pseudomonas spp.in the Rhizosphere of Salt-Stressed Canola

When exposed to biotic or abiotic stress conditions, plants produce ethylene from its immediate precursor 1-aminocyclopropane-1- carboxylate （ACC）, leading to retarded root growth and senescence. Many plant growth-promoting rhizobacteria contain the enzyme ACC deaminase and this enzyme can cleave ACC to form a-ketobutyrate and ammonium, thereby lowering levels of ethylene. The aim of this study was to isolate and characterize ACC deaminase-producing bacteria from the rhizosphere of salt-stressed canola （Brassica napus L.）. Out of 105 random bacterial isolates, 15 were able to utilize ACC as the sole source of nitrogen. These 15 isolates were also positive for indole acetic acid （IAA） production. Phylogenetic analysis based on partial 16S rDNA sequences showed that all isolates belonged to fluorescent Pseudomonas spp. In the canola rhizosphere investigated in this study, Pseudomonas fluorescens was the dominant ACC deaminase-producing species. Cluster analysis based on BOX-AIR-based repetitive extragenic palindromic-polymerase chain reaction （BOX-PCR） patterns suggested a high degree of genetic variability in ACC deaminase-producing P. fluorescens strains. The presence of indigenous ACC-degrading bacteria in the rhizosphere of canola grown in saline soils indicates that these bacteria may contribute to salinity tolerance.

Molecular Characterization of <i>Pseudomonas</i>spp. Isolated from Root Nodules of Various Leguminous Plants of Shekhawati Region, Rajasthan, India

Plant growth promontory Pseudomonas strains were isolated from root nodules of five plant species, viz., Trifolium pretense, Cicer arietinum, Amaranthus polygamus, Vigna mungo, and Trigonella foenum;that plants were denizen of Shekhawati region of Rajasthan. A total of 8 bacterial isolates were evaluated for growth promotion using PGP properties. Partial 16S rDNA sequencing data showed that these 8 bacterial isolates belonged to genus Pseudomonas. MEGA 4.0.2, software was used to construct a neighbor joining tree by employing boot strap method. Result exhibited significant diversity among recovered Pseudomonas strains.

Study on Distribution of Four Pseudomonas Species in Living Environment Using Multiplex PCR

Purpose: The genus Pseudomonas is a ubiquitous microorganism frequently detected from immunocompromised patients. The inherent resistance to numerous antimicrobial agents contributes to the opportunistic character of this pathogen exhaustive monitoring of this pathogen is considered of critical importance to public health organizations. The reliable identification method able to distinguish genetic close Pseudomonas species is needed, because these organisms are difficult to differentiate by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect four Pseudomonas species which are frequently detected from the human oral cavities, and to investigate the distribution of these organisms in the living environment using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the rpoD gene of four Pseudomonas species. Swab samples were collected from fifty washstands, and the distribution of Pseudomonas species was investigated using a conventional PCR at genus level and a multiplex PCR at species level. Results: Multiplex PCR method developed in this study was able to distinguish four Pseudomonas species clearly. The genus Pseudomonas was detected from all samples (100%), whereas P. putida, P, aeruginosa, P. stutzeri and P. fluorescens were detected at 44%, 8%, 4% and 2% in fifty swab samples, respectively. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction. It was indicated that washstands were the uninhabitable environment for P. putida, P, aeruginosa, P. stutzeri and P. fluorescens.

Genetic Diversity of Jute Mallow (Corchorus spp.) Accessions Based on ISSR Markers

Jute mallow is a nutritious leafy vegetable. The leaves are rich in proteins, vitamins and essential amino acids. Molecular characterization of Jute mallow with focus on improvement of leaf yield is scarcely reported. In the present study, inter sequence simple repeats (ISSR) molecular markers were employed to assess genetic diversity and relationships of 83 accessions of Jute mallow from different parts of Africa and Asia conserved at the World Vegetable Center East and Southern Africa. A total of 89 bands were amplified by 8 ISSR primers. Number of polymorphic bands per primer ranged from 2 to 6 with an average of 2.75 bands per primer. Polymorphic information content (PIC) values ranged from 0.390 to 0.760 with average of 0.53. Average Nei’s gene diversity (h) and Shannon’s information index (I) were 0.335 and 0.494 respectively. The highest pairwise genetic distance was 0.431 observed in a population from East Africa accessions. PC1 and PC2 axis explained 21.69% and 11.66% of the total variation respectively. UPGMA cluster analysis grouped the accessions into six main clusters at genetic similarity coefficient of 0.53 as standard value for classification. These results have important implications for jute mallow breeding and conservation.

Screening and pilot-scale evaluation of a highly efficient pesticide-degrading Pseudomonas sp.strain BL5

The widespread use of pesticides has caused serious harm to ecosystems,necessitating effective and environmentally friendly treatment methods.Bioremediation stands out as a promising approach for pollutant treatment,wherein the metabolic activities of microorganisms can transform toxic pesticides into compounds with lower or no toxicity.In this study,we obtained eight pesticide-degrading strains from pesticide-contaminated sites through continuous enrichment and screening.Four highly efficient pesticide-degrading strains(degradation ratios exceeding 80%)were identified.Among them,Pseudomonas sp.BL5 exhibited the strongest growth(exceeding 10^(9) CFU·ml^(-1))and outstanding degradation of benzene derivatives and chlorinated hydrocarbons at both laboratory and pilot scales,with degradation ratios exceeding 98%and 99.6%,respectively.This research provides new tools and insights for the bioremediation of pesticide-related pollutants.

Prevalence and Antibiotic Resistance of Urinary Tract Pathogens, with Molecular Identification of Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp., Using Multiplex Real-Time PCR

Urinary tract infections (UTIs) caused by uropathogens are a significant public health problem, and their treatment primarily relies on antibiotic therapy. However, the increasing global development of antibiotic resistance necessitates updating diagnostic techniques to ensure higher sensitivity and specificity, especially with advancements in science and medicine. This study aimed to evaluate the prevalence of UTIs and antibiotic resistance profiles through urine culture, as well as to identify Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp. in urine samples using a molecular approach with multiplex real-time PCR. From May 3 to July 25, 2023, at the Pietro Annigoni Biomolecular Research Center (CERBA) and Saint Camille Hospital of Ouagadougou (HOSCO), 209 urine samples collected from patients with suspected UTIs were analyzed using both urine culture and multiplex real-time PCR. Among the 209 patients, 52.15% were male and 47.85% female, with an average age of 46.87 ± 21.33 years. Urine cultures revealed an overall UTI prevalence of 23.44%, with a prevalence of 8.13% in men versus 15.31% in women (P = 0.023). The bacterial prevalence rates were as follows: Escherichia coli (12.92%), Klebsiella spp. (7.18%), Enterobacter cloacae (1.44%), Staphylococcus aureus (0.96%), and other bacteria. Klebsiella spp. demonstrated 100% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, while Escherichia coli showed 96.2% and 65.4% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, respectively. PCR analysis of the target bacteria revealed mono-infection prevalence rates of Klebsiella pneumoniae (10.39%), Klebsiella oxytoca (7.79%), and Acinetobacter spp. (7.79%), along with a co-infection prevalence rate of Klebsiella pneumoniae/Acinetobacter spp. (1.30%). This study demonstrated that PCR, with its high sensitivity and specificity, could effectively distinguish Klebsiella pneumoniae from Klebsiella oxytoca and detect Acinetobacter spp. in less than 24 hours—something urine culture alone could not achieve. The relative ease of automating urine PCR testing, combined with its diagnostic accuracy and rapid turnaround time, makes it a valuable addition to modern medical practice for the laboratory diagnosis of UTIs.

Effect of dissolved organic nitrogen on the bloom of Prorocentrum donghaiense and Karenia spp. in the East China Sea coastal waters

Understan ding the mechanism of harmful algal bloom formation is vital for effectively preventing algal bloom outbreaks in coastal environments.Karenia spp.blooms in the East China Sea show a significant correlation with nutrient regimes.However,the impact of key components of nutrients,especially dissolved organic nitrogen(DON),on the blooms of Karenia spp.is not clear.Quantitative research is still lacking.In this study,the cruise observations,field mesocosm-flask culture experiments,and a multinitrogen-tri-phytoplankton-detritus model(NTPD) are combined to reveal the quantitative influence of nutrient regimes on the shift of Prorocentrum donghaiense and Karenia spp.in the East China Sea.It has a synchronism rhythm of diatom-P.donghaienseKarenia spp.-diatom loop in the field culture experiment,which is consistent with the results of the cruise observation.The results showed that the processes of terrigenous DON(TeDON) and dissolved inorganic nitrogen(DIN:NO_(3)^(-)-N,NH_(4)^(+)-N) absorption promoted P.donghaiense to become the dominant algae in the community;whereas the processes of DON from P.donghaiense absorption promoted Karenia spp.to become the dominant algae in ambient DIN exhaustion.In addition,the three-dimensional fluorescence components of humus C,tyrosine and fulvic acid can indicate the processes of growth and extinction of P.donghaiense and Karenia spp.,respectively.This study infers that P.donghaiense and Karenia spp.regime shift mechanism associated with the nutrient regime in coastal waters,which provides a scientific basis for the environmental management of coastal eco system health.

Microbiologically influenced corrosion resistance enhancement of coppercontaining high entropy alloy Fe_(x)Cu_(1−x)CoNiCrMn against Pseudomonas aeruginosa

To enhance the microbiologically influenced corrosion(MIC)resistance of FeCoNiCrMn high entropy alloy(HEAs),a series of Fe_(x)Cu_((1−x))CoNiCrMn(x=1,0.75,0.5,and 0.25)HEAs were prepared.Microstructural characteristics,corrosion behavior(morphology observation and electrochemical properties),and antimicrobial performance of Fe_(x)Cu_((1−x))CoNiCrMn HEAs were evaluated in a medium inoculated with typical corrosive microorganism Pseudomonas aeruginosa.The aim was to identify copper-containing FeCoNiCrMn HEAs that balance corrosion resistance and antimicrobial properties.Results revealed that all Fe_(x)Cu_((1−x))CoNiCrMn(x=1,0.75,0.5,and 0.25)HEAs exhibited an FCC(face centered cubic)phase,with significant grain refinement observed in Fe_(0.75)Cu_(0.25)CoNiCrMn HEA.Electrochemical tests indicated that Fe_(0.75)Cu_(0.25)CoNiCrMn HEA demonstrated lower corrosion current density(i_(corr))and pitting potential(E_(pit))compared to other Fe_(x)Cu_((1−x))CoNiCrMn HEAs in P.aeruginosa-inoculated medium,exhibiting superior resistance to MIC.Anti-microbial tests showed that after 14 d of immersion,Fe_(0.75)Cu_(0.25)CoNiCrMn achieved an antibacterial rate of 89.5%,effectively inhibiting the adhesion and biofilm formation of P.aeruginosa,thereby achieving resistance to MIC.

Strawberry (Fragaria spp.): Cultivation, Production, Consumption, and Marketing in Cameroon

Strawberry (Fragaria spp.) is one of the most important fruits classified as exotic fruits imported into Cameroon. To have an inventory of its cultivation in Cameroon, a survey study was carried out among eight farms of Fragaria spp. from January 2021 to February 2022. The plant was introduced in Cameroon in 2018. There are 13 varieties of Fragaria spp. currently cultivated. Among these 13 varieties, eleven are hybrids of Fragaria x ananassa (“Amiga”, “Amine”, “Camarosa”, “Chandler”, “Charlotte”, “Elsanta”, “Gariguette”, “Madame Moutot”, “Ostara”, “Ruby gem” and “San Andreas”), and two of the hybrids of Fragaria vesca (“Maestro” and “Mara des bois”). The cropping system, irrigation system, and type of fertilizers applied differ from one strawberry farm to another. Biofertilizers (such as mycorrhizal), inorganic and organic fertilizers are actually used to improve production. The potential annual production of strawberries from January 2021 to February 2022, estimated based on the survey data, was 21.216 tons for all growers. Among these eight production farms, the Lolodorf BIO Farm presents 6000 kg (six tons) of strawberries and 100,000 stolons (seedlings) produced, from seven varieties of Fragaria spp. cultivated, with 6 varieties which are hybrids variety Fragaria x ananassa (“Amiga”, “Amine”, “Chandler”, “Gariguette”, “Madame Moutot”, and “Ruby gem”), and one which is a hybrid of Fragaria vesca (“Mara des bois”). Certain diseases were also observed and recorded depending on the growing areas.

Antimicrobial Resistance of Aeromonas spp.and Vibrio spp.Isolated from Fresh Pangasius Fish in Cambodia

The study was conducted to identify Aeromonas spp.and Vibrio spp.from fresh Pangasius fish(n=153)in Cambodia and test their antimicrobial susceptibility to antibiotics.The samples were collected from different wet markets of Phnom Penh city and Kampong Thom,and Siem Reap provinces.The bacteria were isolated by using selective medium and their AMR(Antimicrobial Resistance)profile was tested by API 20E technique,respectively.Susceptibility profile was determined for seven antibiotics commonly used.The Vibrio spp.(34.64%,n=53)was found to be higher than Aeromonas spp.(24.83%,n=38).Four Vibrio and four Aeromonas species were identified where V.parahaemolyticus(57%,n=30)was the highest,followed by V.cholerae(38%,n=20),V.fluvialis(3.8%,n=2)and V.aglinolyticus(1.9%,n=1),whereas A.hydrophila(47%,n=18)was the highest,followed by A.hydrophila/caviae(45%,n=17),A.sobria(5%,n=2),A.caviae(2.6%,n=1).All the species presented high multi-resistance to the tested antibiotics.The antibiotic susceptibility profile to ampicillin(74%-100%),ciprofloxacin(7%-100%),sulfamethoxazole/trimethoprim(14%-100%),florfenicol(14%-100%),oxytetracycline(7%-100%),erythromycin(10%-100%)and colistin sulphate(33%-100%)was revealed resistance level in Aeromonas spp.whereas few species of Vibrio spp.resistant to ampicillin(43%-100%),ciprofloxacin(14%-100%),sulfamethoxazole/trimethoprim(14%-100%),florfenicol(14%-100%),oxytetracycline(20%-100%),erythromycin(29%-100%),colistin sulphate(33%-100%)were also identified.The results revealed these Aeromonas spp.and Vibrio spp.are potentially reservoirs of antibiotic resistance genes.MDR(Multidrug Resistance)was widespread among the samples isolated.That is a high-risk source of contamination since those pathogens and antimicrobials are often used.Our findings highlight that the aquatic environment and fresh Pangasius fish act as reservoirs of AMR Aeromonas spp.and Vibrio spp.which underline the need for a judicious use of antimicrobials and timely surveillance of AMR in aquaculture.Overall,the findings of our study indicated the presence of A.hydrophila,A.hydrophila/caviae,A.caviae,A.sobria,V.parahaemolyticus,V.cholerae,V.alginolyticus and V.fluvialis and high MDR.This result will allow us to identify the potential risk over circulating isolates in animal health and public health and the spread through the food chain offering supports for appropriate sanitary actions.

Botulinum toxin type A-targeted SPP1 contributes to neuropathic pain by the activation of microglia pyroptosis

BACKGROUND Neuropathic pain(NP)is the primary symptom of various neurological condi-tions.Patients with NP often experience mood disorders,particularly depression and anxiety,that can severely affect their normal lives.Microglial cells are as-sociated with NP.Excessive inflammatory responses,especially the secretion of large amounts of pro-inflammatory cytokines,ultimately lead to neuroinflam-mation.Microglial pyroptosis is a newly discovered form of inflammatory cell death associated with immune responses and inflammation-related diseases of the central nervous system.METHODS Two models,an in vitro lipopolysaccharide(LPS)-stimulated microglial cell model and a selective nerve injury model using BTX-A and SPP1 knockdown treatments,were used.Key proteins in the pyroptosis signaling pathway,NLRP3-GSDMD,were assessed using western blotting,real-time quantitative polymerase chain reaction,and immunofluorescence.Inflammatory factors[interleukin(IL)-6,IL-1β,and tumor necrosis factor(TNF)-α]were assessed using enzyme-linked immuno-sorbent assay.We also evaluated microglial cell proliferation and apoptosis.Furthermore,we measured pain sensation by assessing the delayed hind paw withdrawal latency using thermal stimulation.RESULTS The expression levels of ACS and GSDMD-N and the mRNA expression of TNF-α,IL-6,and IL-1βwere enhanced in LPS-treated microglia.Furthermore,SPP1 expression was also induced in LPS-treated microglia.Notably,BTX-A inhibited SPP1 mRNA and protein expression in the LPS-treated microglia.Additionally,depletion of SPP1 or BTX-A inhibited cell viability and induced apoptosis in LPS-treated microglia,whereas co-treatment with BTX-A enhanced the effect of SPP1 short hairpin(sh)RNA in LPS-treated microglia.Finally,SPP1 depletion or BTX-A treatment reduced the levels of GSDMD-N,NLPRP3,and ASC and suppressed the production of inflammatory factors.CONCLUSION Notably,BTX-A therapy and SPP1 shRNA enhance microglial proliferation and apoptosis and inhibit microglial death.It improves pain perception and inhibits microglial activation in rats with selective nerve pain.

Thoracic spine infection caused by Pseudomonas fluorescens:A case report and review of literature

BACKGROUND The clinical incidence of spinal infection is gradually increasing,and its onset is insidious,easily leading to missed diagnosis and misdiagnosis,which may lead to serious complications such as nervous system dysfunction,spinal instability and/or deformity,and cause a huge burden on society and families.Early identification of the causative agent and precision medicine will greatly reduce the suffering of patients.At present,the main pathogenic bacteria that cause spinal infection are Staphylococcus aureus,Streptococcus,Pneumococcus,Escherichia coli,and Klebsiella.There are no reports of spinal infection caused by Pseudomonas fluorescens.CASE SUMMARY We report a 32-year-old female patient with spinal infection.She presented with flank pain,initially thought to be bone metastases or bone tuberculosis,and had a family background of tumors.Her clinical features and changes in imaging and laboratory tests led to the suspicion of thoracic spine infection.Histopathology of the lesion showed inflammation,tissue culture of the lesion was negative several times,and the possible pathogen-Pseudomonas fluorescens was found after gene sequencing of the lesion.The patient recovered completely after a full course of antibiotic treatment.CONCLUSION This report increases the range of pathogens involved in spinal infections,highlights the unique advantages of gene sequencing technology in difficult-todiagnose diseases,and validates conservative treatment with a full course of antibiotics for spinal infections without complications.

Molecular investigation of exoU and exoY virulence genes in Pseudomonas aeruginosa collected from hospitalized patients in North of Iran:A descriptive-analytical study

Objective:To investigate the frequency of exoU and exoY genes in patients with Pseudomonas aeruginosa infection.Methods:In this study,100 clinical isolates of Pseudomonas aeruginosa were collected from patients hospitalized in educational-therapeutic hospitals and were identified using standard microbiological tests.Then,the antibiotic resistance pattern of the isolates was determined by the disk agar diffusion method.The bacterial DNAs were extracted by the alkaline lysis method.Finally,the presence of exoU and exoY genes was evaluated by the PCR test.Results:In this study,47%,72%,29%,39%,40%,and 44%of the isolates were non-susceptible to piperacillin,aztreonam,ceftazidime,imipenem,tobramycin,and ciprofloxacin,respectively.In addition,95%and 93%of the clinical isolates carried the exoU and exoY genes.Blood and fecal isolates had both virulence genes,while only one wound isolate had neither genes.Meanwhile,all urinary isolates contained the exoY gene and only one isolate lacked the exoU gene.Also,88 isolates simultaneously had both exoU and exoY genes.Conclusions:High prevalence of exoU and exoY genes in this region indicates a significant role of typeⅢsecretion system in pathogenesis of Pseudomonas aeruginosa.The typeⅢsecretion system may be a suitable target to reduce the pathogenicity of this bacterium.