[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.

[Value of intravoxel incoherent motion diffusion weighted imaging in evaluating the microstructure changes in white matter hyperintensities].

Objective: To investigate the value of intravoxel incoherent motion diffusion weighted imaging (IVIM DWI) in evaluating microstructure changes in elderly white matter hyperintensities (WMH) patients and to analyze the correlation between IVIM parameters and severity grading and cognitive scores. Methods: Sixty-two WMH patients in Zhejiang Hospital were collected from December 2014 to March 2018 and underwent conventional magnetic resonance (MR) plain scan and diffusion weighted imaging with different b values. The age was 60-92(74±10) years with 37 males, 25 females. The severity of WMH was assessed by T(2) fluid attenuated inversion recovery (FLAIR) sequence and Fazekas score,which were divided into two subgroups. Slow diffusion coefficient (D), fast diffusion coefficient (D(*)) and perfusion fraction (f) from IVIM parameters of double exponential model were compared between regions of WMH (deep WMH (DWMH) and periventricular WMH (PWMH)) and surrounding normal white matter (NWM).The Shapiro-Wilk test was used for normality tests, Kruskal-Wallis tests and Dwass-Steel-Critchlow-Fligner (DSCF) procedure were used for the comparison among these parameters. Furthermore, Wilcoxon two-sample test was used for the comparisons between different severity. Pearson correlation analysis was performed to determine whether these D, D(*), f values were correlated with the mini mental state examination (MMSE) scores. Results: D(D)WMH (0.83(0.72,0.99)×10(-3) mm(2)/s), D(PWMH)((1.13±0.25)×10(-3) mm(2)/s) were significantly higher than D(NWM) ((0.71±0.05)×10(-3) mm(2)/s)(P<0.01). f (DWMH) ((8.94%(7.46%,11.67%)), f (PWMH)(8.34%(6.73%,9.96%)) were significantly higher than f (NWM)(6.71%±1.72%)(P<0.01).D in DWMH were significantly lower than that in PWMH(P<0.01), there's no statistically difference between other groups. D in severe WMH (both DWMH and PWMH) were significantly higher than that in mild WMH (P=0.000 1, P=0.04). Only f in PWMH were positively associated with the MMSE scores (r=0.326 5,P<0.05). Conclusions: IVIM DWI can noninvasively assess the variation of microstructure diffusion and perfusion in WMH in one sequence,which may objectively reflect the severity of these lesions. This method has important clinical significance for better assessment and management of this disease.

[Antimicrobial resistance and molecular epidemiology of foodborne Yersinia enterocolitica in Pudong New District, Shanghai].

Objective: To investigate the antimicrobial resistance and molecular epidemiology of foodborne Yersinia (Y.) enterocolitica in Pudong New District of Shanghai. Methods: Four kinds of raw food samples were collected in retail circulation sites in Pudong from 2012 to 2016. Cold enrichment method was used to isolate Y. enterocolitica and further detection of biotype, serotype, virulent genes, antimicrobial susceptibility of the isolates and pulsed field gel electrophoresis (PFGE) were conducted. Results: A total of 3 900 raw food samples were collected during this period, including poultry product (n=590), livestock product (n=1 074), aquatic product (n=1 488), vegetable (n=748), in which 111 (2.8%) were contaminated by Y. enterocolitica. The detection rates of Y. enterocolitica in poultry product samples (5.3%, 31/590) and livestock product samples (4.5%, 48/1 074) were higher than those in aquatic product samples (1.6%, 24/1 488) and vegetable samples (1.1%, 8/748). The predominant biotype was 1A (95.5%) and predominant serotype was O∶8 (42.3%). All the strains lacked ail, ystA, yadA and virF genes, which encoded pathogenic Y. enterocolitica. Seventy six (68.5%) strains harbored ystB gene, in which 35 (31.5%) belonged to 1A/O∶8/ystB pattern. Most strains were resistant to ampicillin (74.8%) and amoxicillin/clavulanic acid (70.3%), and non-sensitive rate to Cefoxitin was over 50.0%. No third generation cephalosporin or fluoroquinolone resistant strains were detected, but 38.7% (43/111) strains were multidrug resistant (MDR). Serotype O∶8 and O∶5 strains had 44 and 18 PFGE patterns, respectively. Conclusions: The main foodborne exposure sources of Y. enterocolitica in raw food were poultry and livestock products in Pudong New District. 1A/O∶8/ystB was the predominant pattern with potential pathogenicity despite lacks of typical pathogenic virulent genes. The antimicrobial resistant rates of Y. enterocolitica were at a low level, but MDR strains still existed. Molecular types of the isolates showed highly genetic diversity.

MiR-638 serves as a tumor suppressor by targeting HOXA9 in glioma.

OBJECTIVE: MiR-638 is constantly downregulated and serves as a tumor suppressor in various cancers. Its role in gliomas remains unclear. This study is designed to investigate the clinical significance and the pathogenic role of miR-638 in human gliomas. PATIENTS AND METHODS: Quantitative Real-time PCR was performed to analyze the expression of miR-638 in the tumor and adjacent tissues of 24 glioma patients. The association between the expression of miR-638 and clinical features were examined. Survival of patients was studied by Kaplan-Meier curves. The impact of miR-638 on cell growth and apoptosis was determined by CCK-8 assay, colony formation assay, cell cycle analysis and Annexin V-FITC-PI apoptosis assay. The effect of miR-638 on HOXA9 was determined by luciferase assay and Western blot. The effect of miR-638 and HOXA9 on expression of oncogenes, Cyclin D1 and C-MYC was determined by Western blot. RESULTS: MiR-638 expression was constantly downregulated in glioma tumor tissue, which is negatively correlated with the WHO grade. MiR-638 expression was associated with clinical features such as tumor size, KPS score and WHO grade. Patients with low miR-638 had a worse overall survival than those with high expression. Experimentally, miR-638 directly targeted HOXA9 to suppress its expression, leading to attenuations of cell proliferation, colony formation and cell cycle progression and enhanced basal apoptosis level. MiR-638/HOXA9 axis also suppressed the expression of Wnt/beta-catenin-regulated oncogenes, Cyclin D1 and C-MYC. CONCLUSIONS: MiR-638 is a constantly downregulated microRNA in gliomas and is associated with its prognosis. MiR-638 regulates cellular malignancy of gliomas through targeting HOXA9. Thus, miR-638/HOXA9 signaling axis may have therapeutic potential in gliomas.

[The concomitant gene alterations impact the therapeutic efficacy of epidermal growth factor receptor-tyrosine kinase inhibitors in advanced non-small cell lung cancer patients with epidermal growth factor receptor sensitive mutation].

Objective: To investigate if concomitant gene alterations impact the therapeutic efficacy of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKI) in advanced non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) sensitive mutation. Methods: From November 2016 to December 2017, 51 patients (19 males and 32 females, age 37-85 years) with histology or cytology diagnosed,locally advanced or metastatic NSCLC from Zhongshan Hospital Fudan University were prospectively recruited in the study. All patients harboring EGFR sensitive mutation detected by a 123 lung cancer specific gene panel of next-generation sequencing(NGS) analysis were under treatment of EGFR-TKIs. Multi-factors analysis of the correlation between EGFR-TKIs efficacy and concomitant gene alterations were analyzed by multivariate Cox regression model. Results: 82% of the NSCLC patients with EGFR mutation presented concomitant gene alterations with an average number of 2.1. Patients not harboring concomitant gene alterations had a longer median progression free survival (mPFS: not reached vs 8.8 m, P=0.008). Those who had less than 2 concomitant genes had a higher objective response rate[ORR: 52% (17/33) vs 33% (6/18) , P=0.251]and better mPFS (13.8 vs 8.0 m, P=0.003). The top 3 concomitant gene alterations were TP53 gene mutation(55%, 28/51), EGFR gene amplification (26%, 13/51) and RB1 gene mutation (18%,9/51) respectively. The mPFS of EGFR-TKI treatment in patients with either one of these 3 concomitant genes was 8.0, 8.0, and 6.0 months respectively, significantly shorter than those without one of the 3 gene alterations (13.8, 13.1, and 10.8 months respectively). Multivariate Cox regression revealed that concomitant gene abnormalities (P=0.036) and accompanied by RB1 gene mutation (P=0.025) were independent risk factors for the survival benefit of EGFR-TKI in the treatment of advanced NSCLC with EGFR-sensitive mutation. Conclusions: The efficacy of EGFR-TKI decreased significantly in advanced NSCLC with EGFR-sensitive mutation who had concomitant gene abnormalities, especially accompanied by more than 2 of the 3 gene alterations (TP53 gene mutation, EGFR gene amplification or RB1 gene mutation). This study suggested that the concomitant gene alterations should be an important issue for consideration when applying a personalized combination therapy for advanced NSCLC harboring EGFR sensitive mutation.

Clinical study on the early application and ideal dosage of urokinase after surgery for hypertensive intracerebral hemorrhage.

OBJECTIVE: The objective of the present study was to investigate the clinical efficacy of early application and ideal dosage of urokinase after minimally invasive surgery for hypertensive intracerebral hemorrhage (HICH). PATIENTS AND METHODS: We consecutively enrolled 132 patients with HICH who underwent CT-guided stereotactic intubation of the hematoma combined with dissolution using urokinase, where the urokinase was injected at 24 h after intubation. The 40 patients in the low-dosage group received 10-30 thousand units of urokinase; the 46 patients in the moderate-dosage group received 40-60 thousand units of urokinase; and the 46 patients in the high-dosage group received 70-100 thousand units of urokinase. After the drainage tubes were clamped for 4-6 h, clamps were removed for drainage for 24 h, and the intubated tubes were maintained for 3-5 d. Patients in all groups were followed up for 6 months, and the clinical outcomes were compared. RESULTS: The clearance of hematomas in the high-dosage group was significantly improved compared with the other two groups (p<0.05), and the occurrence rates of complications in the moderate- and high-dosage groups were significantly higher than in the low-dosage group (p<0.05). During follow-up, the Chinese stroke scale and Barthel index scores in the high-dosage group were higher than those in the other two groups (p<0.05). The serum levels of matrix metalloproteinase-9 (MMP-9) and neuron-specific enolase (NSE) in the high-dosage group were lower than those in the other two groups (p<0.05). CONCLUSIONS: CT-guided stereotactic intubation of the hematoma combined with dissolution using urokinase is effective for eliminating the hematoma for treatment of HICH with few complications. For recovery of neurological functions and improvement of regular life skills, it is considered to be associated with decreases in the serum levels of MMP-9 and NSE.

Comparison of target-controlled infusion and manual infusion for propofol anaesthesia in children.

BACKGROUND: One major criticism of prolonged propofol-based total i.v. anaesthesia (TIVA) in children is the prolonged recovery time. As target-controlled infusion (TCI) obviates the need to manually calculate the infusion rate, the use of TCI may better match clinical requirements, reduce propofol dose, and shorten recovery time. METHODS: Children of ASA grade 1, aged 1-12 yr, were recruited and randomly assigned to TCI or manual infusion. Children in the TCI group had propofol delivered by TCI. Children for manual infusion had a loading dose of 2.5 mg kg-1 with subsequent infusion rates of 15, 13, 11, 10, and 9 mg kg-1 h-1. Attending anaesthesiologists adjusted the propofol dosage to keep the Bispectral Index™ (BIS) between 40 and 60. RESULTS: Seventy-four children completed the study. The time taken to extubate the trachea after cessation of propofol was 15.1 (5.5) and 16.2 (6.1) min for children who had TCI and manual infusion, respectively (P=0.42). The mean propofol infusion rate was 16.7 [standard deviation (sd) 4.2] mg kg-1 h-1 in the TCI group and 14.6 (3.1) mg kg-1 h-1 in the manual infusion group (P=0.036). The percentage of time when BIS was >60 was significantly lower in the TCI than the manual infusion group [10.2% (18.4%) vs 23.2% (26.3%), P=0.016]. DISCUSSION: Use of TCI led to higher propofol doses but not prolonged recovery time in children compared with manual infusion. It was associated with a greater percentage of time when the BIS was in the desired range and it may be an easier method for titration of propofol administration during anaesthesia or sedation. CLINICAL TRIAL REGISTRATION: ChiCTR-IOD-16010147.

[Treatment of single one-stage posterior atlantoaxial fixation in Chiari malformation].

Objective: To explore the effects of surgical technique of single one-stage posterior C(1-2) screw rod fixation of Chiari malformation (CM) associated with occipitalization and without atlantoaxial dislocation. Methods: A total of 23 patients with CM treated between January 2014 and October 2015 in Department of Neurosurgery of Chinese People's Liberation Army General Hospital were retrospective reviewed. All of them were diagnosis with CM associated with occipitalization and without atlantoaxial dislocation, including 8 males and 15 females, aging from 11 to 57 years (mean (35.5±10.52) years). Single one-stage posterior C(1-2) screw rod fixation with bone grafting fusion was performed. Operation time and intraoperative blood loss were recorded. Japanese Orthopaedic Association (JOA) scores and Odom rating were used to evaluate the clinical effects at pre- and post-operative. Regression of the cerebellar tonsillar was measured by MRI. The results were analyzed by paired samples t test. Results: Twenty-three patients were implanted screws successfully, the vertebral artery injury and cerebrospinal fluid leakage were not found. The mean operation time was (172.7±19.9) minutes, the intraoperative blood loss was (153.9±49.3) ml. Compared to preoperative, the JOA score increased (13.7±1.6 vs. 11.5±1.4) and the tonsillar herniation decreased ((0.8±0.6)cm vs. (1.9±0.6) cm) in the last follow-up, there were statistical difference (t=13.386, P<0.01; t=17.995, P<0.01). The results of the postoperative Odom grading were as follows: 6 cases were perfect (26.1%), 13 cases were good (56.5%), 4 cases were moderate (17.4%) and no case was poor.No signs of instrument loosen or screw broken was noticed. 100% bony fusion rate was achieved. The follow-up time was 6 to 23 months (mean (10.5±3.2) months). One case developed internal fixator related discomfort, the symptom was relieved by internal fixator removal surgery performed 4 months after the operation when osseous fusion had already been achieved. No new neurologic symptoms were observed in other 22 patients. Conclusions: The results of the study substantiates the effectiveness of single one-stage posterior fixation strategy for CM, which is associated with occipitalization and without atlantoaxial dislocation. This technique could be an alternative choice for this type of CM.

[Study of epidemic area on Tsutsugamushi disease in Taizhou from 2013 to 2014].

Objective: To study the epidemiological characteristics of tsutsugamushi disease, and to confirm the existence of the disease's epidemic foci in Taizhou. Methods: From 2013 to 2014, Dongxing town hospital and Xingqiao town hospital were selected as specimen collection sites in Jingjiang city. Blood samples (5 ml) were collected from 40 patients with acute tsutsugamushi disease. A total of 59 rodents were captured with cage night method in the survey sites at 5, 7, 9, 10, and 11 months in 2013, from which, the spleen, liver, and kidney specimens were selected. Chigger mites were captured by small blackboard method and from the ears of the captured rodents. A total of 226 small blackboards were laid, 27 mites were captured, and the samples were grounded into suspension. Nested-polymerase chain reaction and cell and tissue culture techniques were used to test the specimen from the probable patients, host animals and chigger mites. Results: Among the 40 acute tsutsugamushi disease blood samples, 29 were found to meet the test requirements, 17 were positive for orientia tsutsugamushi nucleic acid with 59% of the positive rate, and 1 stran orientia tsutsugamushi was isolated. 59 rats were captured and the density of mice was 5.5%. Among them, there were 26 Mus musculus (2.4%), 18 Rattus flavipectus (1.7%) and 15 Smelly shrew (density 1.4%). 1 Smelly shrew was tested positive for orientia tsutsugamushi nucleic acid, and the negative results were found in the other rodent specimens. 27 Chigge mites were collected by small blackboard method and the density of mites was 0.12 for each blackboard, among which 3 larvae and 24 nymphs were found. 33 Chigger mites were collected from the ears of 3 Smelly shrew, and the density of the mite was 11 per mouse. All the captured Chigger mites were identified as Leptotrombidium scutellare and 1 group of specimens of Chigger mites from the external environment were positive for orientia tsutsugamushi nucleic acid. Conclusion: There was a high density of mice in the epidemic area from May to November and the species of the chigger mites were Chigger mites in Taizhou. The nucleic acid of the oriental tsutsugamushi was detected in the patients with acute scrub typhus, rodents and vectors. According to the above-mentioned results, it was considered that the scrub typhus epidemic area of Taizhou city has the natural foci of scrub typhus.

[Clinical epidemiology and antimicrobial resistance of nontyphoidal Salmonella enteric infections in children: 2012-2014].

OBJECTIVE: To understand the clinical epidemiology and manifestation of nontyphoidal Salmonella (NTS)enteric infections and serotypes and antibiotic resistance patterns of NTS in children. METHOD: All cases with probable acute bacterial inflammatory diarrhea were included into this study from the Children's Hospital of Fudan University during January 2012 through December 2014. Salmonella species and other major enteric bacteria were routinely isolated from the fresh stool sample of the enrolled patients at their first visit to the enteric clinic. Salmonella isolates were serotyped and tested for antimicrobial susceptibility testing using Kirby-Bauer method at the reference laboratory of Shanghai Municipal Center for Disease Control and Prevention. All enrolled cases had clinical records. The enumeration data were described by constituent ratio. RESULT: Of the enrolled 3 143 patients, 742 (23.6%) were confirmed to have Salmonella infections, and NTS was identified in 738 (23.5%) cases and Salmonella Paratyphi B was identified in 4 (0.1%) cases, respectively. The isolation rate of NTS exceeded diarrheagenic Escherichia coli (6.7%, 182/2 727), Campylobacter (4.1%, 129/3 143) and Shigella (2.4%, 77/3 143). Nine serogroups and 41 serovars were identified in 742 salmonella isolates. Salmonella Enteritidis and Salmonella Typhimurium were the most common serovars, representing 42.6% (316/742) and 28.2% (209/742) of isolates, respectively.NTS infections were prevalent year-round with a seasonal peak from May to October. The common symptoms included fever (57.5%, 424/738) and blood-in-stool (31.0%, 229/738). Antimicrobial susceptibility showed only 8 (1.1%) of 742 isolates sensitive to 16 tested antibiotics.Resistance to the current first-line antibiotics such as the third-generation and(or) fourth-generation cephalosporins (cefotaxime and cefepime), ciprofloxacin was detected in 10.0%(74/738), 8.1%(60/738) and 1.1%(8/738), respectively. CONCLUSION: NTS is the leading causative agent responsible for bacterial inflammatory diarrhea in Shanghainese children. The prevalence of NTS enteric infections show an increasing trend year by year. It is essential to continuously monitor the antimicrobial resistance of NTS and trace the source of infection to prevent and treat NTS infections in Shanghai.

Proteomic analysis reveals the mechanisms of Mycena dendrobii promoting transplantation survival and growth of tissue culture seedlings of Dendrobium officinale.

AIMS: Dendrobium officinale is an important traditional Chinese medicinal herb. Its seedlings generally show low survival and growth when transferred from in vitro tissue culture to a greenhouse or field environment. In this study, the effect of Mycena dendrobii on the survival and growth of D. officinale tissue culture seedlings and the mechanisms involved was explored. METHODS AND RESULTS: Mycena dendrobii were applied underneath the roots of D. officinale tissue culture seedlings. The seedling survival and growth were analysed. The root proteins induced by M. dendrobii were identified using two-dimensional (2-D) electrophoresis and matrix-assisted laser desorption/ionization time-of-flight MS (MALDI-TOF-MS). Mycena dendrobii treatment significantly enhanced survival and growth of D. officinale seedlings. Forty-one proteins induced by M. dendrobii were identified. Among them, 10 were involved in defence and stress response, two were involved in the formation of root or mycorrhizae, and three were related to the biosynthesis of bioactive constituents. CONCLUSIONS: These results suggest that enhancing stress tolerance and promoting new root formation induced by M. dendrobii may improve the survival and growth of D. officinale tissue culture seedlings. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides a foundation for future use of M. dendrobii in the large-scale cultivation of Dendrobiums.