[The role of Nrf2 in the alteration of tight junction protein expression in choroid plexus epithelial cells created by lanthanum-activated MMP9].

Objective: To investigate the effect of nuclear factor erythroid 2-related factor 2 (Nrf2) in the alteration of tight junction protein expression in choroid plexus epithelial cells created by lanthanum-activated matrix metalloproteinase 9 (MMP9) . Methods: In October 2020, immortalized rat choroid plexus epithelial cell line (Z310) cells were used as the blood-cerebrospinal fluid barrier in vitro, and were divided into control group and 0.125, 0.25, 0.5 mmol/L lanthanum chloride (LaCl(3)) treatment group. After treating Z310 cells with different concentrations of LaCl(3) for 24 hours, the morphological changes of Z310 cells were observed under inverted microscope, the protein expression levels of MMP9, occludin and zonula occludens-1 (ZO-1) were observed by cellular immunofluorescence method, and the protein expression levels of MMP9, tissue inhibitors of metalloproteinase1 (TIMP1) , occludin, ZO-1 and Nrf2 were detected by Western blotting. The level of reactive oxygen species (ROS) in cells was detected by flow cytometry. Results: Compared with the control group, Z310 cells in the LaCl(3) treatment group were smaller in size, with fewer intercellular junctions, and more dead cells and cell fragments. The expression level of MMP9 protein in cells treated with 0.25 and 0.5 mmol/L LaCl(3) was significantly higher than that in the control group (P<0.05) , and the expression level of TIMP1 and tight junction proteins occudin and ZO-1 was significantly lower than that in the control group (P<0.05) . Compared with the control group, the ROS production level in the 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly increased (P<0.05) , and the Nrf2 protein expression level in the 0.125, 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly decreased (P<0.05) . Conclusion: Lanthanum may increase the level of ROS in cells by down regulating the expression of Nrf2, thus activating MMP9 to reduce the expression level of intercellular tight junction proteins occludin and ZO-1.

[Epidemiological characteristics of Yersinia enterocolitica in patients with diarrhea from 2013 to 2019 in Pudong New Area, Shanghai].

Objective: To investigate the epidemiological characteristics of Yersinia enterocolitica in patients with diarrhea in Pudong New Area, Shanghai. Methods: Active surveillance of diarrhea was conducted in 14 sentinel hospitals (three tertiary-level hospitals, nine secondary-level hospitals, and two primary-level hospitals) from January 2013 to December 2019 in Pudong New Area of Shanghai, China base on their location, catchment area, and patient volume. Cold enrichment method was used to isolate Y. enterocolitica and further detection of bioserotype, virulence genes and antimicrobial susceptibility of the isolates were conducted. The difference of rates was determined using chi-square test or Fisher's exact test. Results: A total of 12 941 diarrhea cases were included, and 0.7% (88/12 941) cases were confirmed with Yersinia enterocolitica infection. 67.0% (59/88) cases were single infection, 33.0% (29/88) cases were mixed infections. Detection rates of Y. enterocolitica increased annually (0.3%-1.2%) and were highest in children<5 years of age (1.1%, 37/3 218) and in spring (1.1%, 32/2 998) (χ2 were 18.64 and 9.76, respectively, P<0.05). 58.0% (51/88) cases had watery diarrhea, 15.9% (14/88) had fever and 14.8% (13/88) had vomiting. The predominant bioserotypes were 3/O:3 (53.4%, 47/88), followed by 1A/O:8 (15.9%, 14/88) and 1A/O:5(6.8%, 6/88), respectively. Bioserotype 3/O:3 counted for the highest proportions (89.2%, 33/37) in children <5 years of age. All the strains of bioserotype 3/O:3 harbored ail, ystA, yadA and virF genes, which encoded pathogenic Y. enterocolitica. 11/14 strain of 1A/O:8 and 4/6 strains of 1A/O:5 harbored ystB gene. Most strains were resistant to ampicillin (80.7%,71/88) and amoxicillin/clavulanic acid (71.6%,63/88), and 63.8% (56/88) strains were multidrug resistance (MDR). The difference of antimicrobial resistance rates between 3/O:3 and non 3/O:3 was statistically significant in ampicillin, cefoxitin, nalidixic acid, tetracycline and ampicillin/sulbactam (χ2 was 14.68, 43.80, 41.86, 30.54 and 5.07, respectively, P<0.05). Conclusion: The detection rate of Yersinia enterocolitica was higher in children than in adults in Pudong New Area, Shanghai. The predominant bioserotype was pathogenic 3/O:3 with multidrug resistance.

[Analysis on serotype and antimicrobial resistance of invasive non-typhoidal Salmonella].

Objective: To investigate the serotypes and antimicrobial resistance of seven invasive non-typhoidal Salmonella (iNTS) isolates. Methods: For 7 iNTS strains collected, serotype identification, antimicrobial susceptibility testing and whole genome sequencing were performed. We identified, annotated and analyzed the serotypes, MLST types, and antimicrobial resistance genes. Results: Among the 7 tested iNTS isolates, we found one Salmonella Typhimurium strain and two Salmonella Ⅰ 4, [5], 12: i:- strains whose MLST types were ST34, two Salmonella Enteritidis strains, one Salmonella Corvallis strain and one strain of unknown serotype with the antigenic formulae of Ⅰ 4, [5], 12: d:- (ST279 type). Six of seven strains were monophasic and the deletion or pseudogenization of Salmonella Flagellum gene might contribute to the enhancement of Salmonella invasiveness. None was found to be resistant to tigarcycline, aztreonam, amikacin, cephalosporins and carbapenem and one Salmonella Typhimurium strain was found to be co-resistant to eight classes of antimicrobials at the same time. Resistance genes were generally in accord with relative resistant phenotypes. Conclusion: The iNTS strains could show high level multi-drug resistance, indicating that close attention should be paid to the resistance of iNTS though the overall resistance might be relatively not high.

Antibiofilm activity against Staphylococcus aureus and content analysis of Taraxacum Officinale phenolic extract.

Taraxacum Officinale, commonly called dandelion, is herbaceous perennial belonging to the family of Asteraceae, having good antibacterial effects which are related to its phenolic substances. In this study, the effect of phenolic contents as well as the antibiofilm activity against Staphylo- coccus aureus of phenolic extract from T. Officinale were evaluated in vitro. With 70% metha- nol-water (v/v) as a solvent, the dandelion was extracted by ultrasonic assisted extraction method. Subsequent identification and quantification of phenol in extract was carried out using High Performance Liquid Chromatography (HPLC). The minimum inhibitory concentration and anti- bacterial kinetic curve of dandelion phenolic extract were analyzed by spectrophotometry. Changes in extracellular alkaline phosphatase (AKP) contents, electrical conductivity, intracellular protein contents, and DNA of S. aureus after the action of dandelion phenolic extract were determined to study its effect on the permeability of S. aureus cell wall and cell membrane. The results showed that chlorogenic acid (1.34 mg/g) was present in higher concentration, followed by lute- olin (1.08 mg/g), ferulic acid (0.22 mg/g), caffeic acid (0.21 mg/g), and rutin (0.19 mg/g) in the dandelion phenolic extract. The minimum inhibitory concentration (MIC) of dandelion phenolic extract against S. aureus was 12.5 mg/mL. The antibacterial kinetic curve analysis showed that the inhibitory effect of dandelion phenolic extract on S. aureus was mainly in the exponential growth phase. After applying the dandelion phenolic extract, the growth of S. aureus was signifi- cantly inhibited entering into the decay phase early. Furthermore, after the action of dandelion, the extracellular AKP contents of S. aureus, the electrical conductivity and the extracellular protein contents were all increased. The phenolic extract also affected the normal reproduction of S. aureus. These results suggest that dandelion has an inhibitory effect on S. aureus, and the mechanism of its action was to destroy the integrity of the cell walls and cell membranes.

Nonequilibrium pattern formation in circularly confined two-dimensional systems with competing interactions.

We numerically investigate the nonequilibrium behaviors of classic particles with competing interactions confined in a two-dimensional logarithmic trap. We reveal a quench-induced surprising dynamics exhibiting rich dynamic patterns depending upon confinement strength and trap size, which is attributed to the time-dependent competition between interparticle repulsions and attractions under a circular confinement. Moreover, in the collectively diffusive motions of the particles, we find that the emergence of dynamic structure transformation coincides with a diffusive mode transition from superdiffusion to subdiffusion. These findings are likely useful in understanding the pattern selection and evolution in various chemical and biological systems in addition to modulated systems, and add a new route to tailoring the morphology of pattern-forming systems.

[Clinical features and outcome of treatment for novel coronavirus pneumonia: a meta-analysis].

Objective: To investigate the clinical features and outcome of treatment for novel coronavirus pneumonia. Methods: Literature on novel coronavirus pneumonia was retrieved from PubMed and EMBASE databases. The relevant data was extracted and a meta-analysis was performed using StatsDirect statistical software V.2.8.0 to calculate the combined odds ratio. Results: Seven studies were included, consisting of 1594 cases. The meta-analysis result showed that the most common clinical symptoms of the novel coronavirus pneumonia were fever (91.6%) and cough (64.5%), followed by dyspnea (32.8%) and sputum (28.1%). Headache (10.5%), sore throat (11.2%), hemoptysis (3.2%), diarrhea (6.6%) and the other symptoms were relatively rare. Aspartate aminotransferase (29%), alanine transaminase (22.7%), and total bilirubin (11.7%) levels were elevated, except for serum albumin levels (80.4%). The common therapeutic agents used were antibiotics (87.7%), antiviral drugs (75.5%), and glucocorticoids (26.6%), while antifungal agents (7.7%) were used in few. Mechanical ventilation (13.4%), extracorporeal membrane oxygenation (1.9%), and continuous renal replacement therapy (3.8%) were used in severe cases. The rate of mortality in hospital was 7.7%, respectively. Heterogeneity between studies was significant; however, subgroup and sensitivity analysis had failed to identify clear sources of heterogeneity. Conclusion: Fever, cough and liver dysfunction are the main clinical manifestations of this disease and the mortality rate is low.

Structural transitions for 2D systems with competing interactions in logarithmic traps.

We propose a confinement model and study numerically the structural properties of particles with competing interactions in logarithmic traps (i.e., the confinement potential is a logarithmic function). A rich variety of cluster structures are observed as a function of trap steepness, trap size, and particle density. In addition to the consistent results with previous studies for a harmonic confinement, we observe some new stable structures, including a hybrid cluster structure consisting of clumps surrounded by a circular stripe, parallel stripes, or homogeneous voids surrounded by a ringlike arrangement of clumps, and a gear-like cluster with fringed outer rims evenly arranged along the circumference. Our work reveals that such self-organized structures arise due to the radial density reconfiguration in a finite confined system corresponding to the unconstrained systems, which is controlled by the interplay between the long-range repulsions and the attractions to the minimum of the confinement potential. Such results are likely relevant in understanding the structural properties of confined mermaid systems.

[The first identification of epidemic clone of enterotoxic Escherichia coli O∶6 serogroup highly associated with azithromycin resistance in Shanghai].

Objective: To investigate the molecular characterization of adult diarrhea cases caused by enterotoxic Escherichia coli (ETEC) and explore the practical model of epidemiology for laboratory technique and data needs based on the surveillance network. Methods: Epidemiological design and sampling targeted adult cases ETEC caused diarrhea in epidemic season. The enterotoxin type, serogroup, resistance, colonization factor and molecular type of ETEC were identified. Multiple dynamic phenotypic characteristics of ETEC were indicated by multidimensional and multivariable data. Results: From 2016 to 2018, 84 eligible ETEC strains were detected. The dominant serums/toxins were O∶6 (STh), O∶25 (LT), O∶159 (STh), O∶153 (STh). O∶6 (STh+CS21), which replaced O∶25 and O∶159 as the popular clones in 2018. Six cases of O∶153 (STh+CFA/I+CS8+PT34) in outbreak in 2017 were imported ones. The resistance rates of ETEC strains detected in adults to sulfasoxazole, naproxinic acid, ampicillin and azithromycin were more than 30%, multidrug resistance (MDR) reached 58.3%. Serum/toxin types suggested that attenuated strains were more likely to become MDR. Molecular typing confirmed that the genetic similarity of the dominant clone of O∶6 serogroup (PT20-24) was higher than O∶25 and O∶159. There was a high correlation between the minimal inhibitory concentration (MIC) of azithromycin and the resistant gene mphA (87.5%, 28/32). O∶6 (STh+CS21+mphA) resistant clone was first detected in 2016. Conclusion: A new epidemic clone in adult ETEC diarrhea cases in Shanghai was O∶6 (STh+CS21+mphA). For the first time the association between azithromycin resistance gene mphA and a serum group of ETEC was observed. Multidimensional and multivariate analysis techniques based on epidemiology can help reveal the potential transmission pattern of ETEC for the accurate surveillance and early warning of outbreaks.

[The hemodynamic study of internal jugular vein in patients with pulsatile tinnitus by color doppler sonography].

Objective:To investigate the hemodynamic changes of internal jugular vein（IJV） in patients with pulsatile tinnitus（PT） detected by color doppler sonography（CDS）. To explore the correlation of PT with IJV drainage dominance and the value of CDS in the diagnosis of PT. Method:Two hundred and twenty-one patients with PT were examined with the CDS and IJV compression test. The hemodynamic data of the upper, middle and lower IJV were compared with both PT side and unaffected side. The correlation of the PT with the IJV drainage dominance was analyzed. Result:A total of 125 cases（56.6%） of 221 patients with PT underwent ultrasonic IJV compression test with positive results, 96 cases（43.3%） with negative results. In the positive compression test group, the tinnitus disappeared after IJV being completely compressed in 109 cases（87.2%）, and in 16 cases（12.8%） after IJV being partial closure. The extent of sectional area reduction was: upper jugular vein（75.41 ±9.39）%, middle jugular vein（80.25±13.16）%, lower jugular vein（86.58± 7.53）%. The blood flow volume on the tinnitus side was obviously higher than that on the unaffected side（P=0.001）. In the negative compression test group, there was no significant difference in blood flow volume between tinnitus side and unaffected side. In the IJV positive compression test group, 54 cases（43.2%） were IJV drainage dominance combined, and 71 cases（56.8%） were non-dominant combined. In the IJV negative compression test group, 24 cases（25%） were IJV drainage dominance combined, and 72 cases（75%） were non-dominant combined. Conclusion:The positive result of IJV compression test using CDS and the flow volume of IJV on the affected side significantly higher than that on the unaffected side contributed to the diagnosis of venous PT. In this study, the IJV non-dominant side was more common in the venous PT.

[The study of pleural effusion supernatant cell-free tumor DNA in tumor mutational burden assessment of advanced lung cancer].

Objective: To evaluate the feasibility of cell-free tumor DNA in pleural effusion supernatant for assessing the tumor mutational burden (TMB) of advanced lung cancers. Methods: From December 2016 to August 2018, 34 lung cancer patients (19 males and 15 females) with pleural effusion were enrolled at Zhongshan Hospital, Fudan University. The median age of the patients was 65 (range, 34-85) years. Before systemic or local antitumor therapy, tumor specific mutations in tumor tissue, pleural effusion supernatant, pleural effusion sediment, and plasma samples from these patients were examined using targeted next-generation sequencing, and TMB levels were calculated respectively. Subgroup analysis was based on smoking history and driver mutation status. Statistical differences were determined using SPSS 16.0 software, and individual groups were compared using the one-way analysis of variance (ANOVA) and LSD-t test. Results: The median TMB level of pleural effusion supernatant was 6.23 mutations/Mb, similar to that of tumor tissue (6.23 vs 6.86 mutations/Mb, t=1.174, P=0.245), but significantly higher than that of pleural effusion sediment (2.49 mutations/Mb, t=3.044, P=0.003) and plasma (2.49 mutations/Mb, t=2.464, P=0.016). Compared with tumor tissue in TMB assessment, pleural effusion supernatant had a positive percentage agreement of 52% (9/17), and a negative percentage agreement of 65% (11/17). Subgroup analysis showed that the TMB level was higher in smokers (n=11) than that in non-smokers (n=23, 14.4 vs 5.4 mutations/Mb, t=3.238, P=0.003). Conclusion: For advanced lung cancer patients with pleural effusion, pleural effusion supernatant is a promising substitute to tumor tissue for TMB assessment, which is a potential biomarker for immunotherapy.