Outer membrane vesicles generated by an exogenous bacteriophage lysin and protection against Acinetobacter baumannii infection.

BACKGROUND: The outer membrane vesicles (OMVs) produced by Gram-negative bacteria can modulate the immune system and have great potentials for bacterial vaccine development. RESULTS: A highly active Acinetobacter baumannii phage lysin, LysP53, can stimulate the production of OMVs after interacting with A. baumannii, Escherichia coli, and Salmonella. The OMVs prepared by the lysin (LOMVs) from A. baumannii showed better homogeneity, higher protein yield, lower endotoxin content, and lower cytotoxicity compared to the naturally produced OMVs (nOMVs). The LOMVs contain a significantly higher number of cytoplasmic and cytoplasmic membrane proteins but a smaller number of periplasmic and extracellular proteins compared to nOMVs. Intramuscular immunization with either LOMVs or nOMVs three times provided robust protection against A. baumannii infections in both pneumonia and bacteremia mouse models. Intranasal immunization offered good protection in the pneumonia model but weaker protection (20-40%) in the bacteremia model. However, with a single immunization, LOMVs demonstrated better protection than the nOMVs in the pneumonia mouse model. CONCLUSIONS: The novel lysin approach provides a superior choice compared to current methods for OMV production, especially for vaccine development.

Calcium-binding motif-mediated binding of redundant calcium offers a chimeolysin enhanced bactericidal activity and extended host range under physiological conditions.

OBJECTIVES: Calcium-binding motifs are shared by multiple bacteriophage lysins; however, the influence of calcium on the enzymatic activity and host range of these enzymes is still not understood. To address this, ClyF, a chimeric lysin with a putative calcium-binding motif, was used as a model for in vitro and in vivo investigations. METHODS: The concentration of calcium bound to ClyF was determined by atomic absorption spectrometry. The influence of calcium on the structure, activity and host range of ClyF was assessed by circular dichroism and time-kill assays. The bactericidal activity of ClyF was evaluated in various sera and a mouse model of Streptococcus agalactiae bacteraemia. RESULTS: ClyF has a highly negatively charged surface around the calcium-binding motif that can bind extra calcium, thereby increasing the avidity of ClyF for the negatively charged bacterial cell wall. In line with this, ClyF exhibited significantly enhanced staphylolytic and streptolytic activity in various sera containing physiological calcium, including human serum, heat-inactivated human serum, mouse serum and rabbit serum. In a mouse model of S. agalactiae bacteraemia, intraperitoneal administration of a single dose of 25 µg/mouse ClyF fully protected the mice from lethal infection. CONCLUSIONS: The present data collectively showed that physiological calcium improves the bactericidal activity and host range of ClyF, making it a promising candidate for the treatment of infections caused by multiple staphylococci and streptococci.

Phage-bacterial evolutionary interactions: experimental models and complications.

Phage treatment of bacterial infections has offered some hope even as the crisis of antimicrobial resistance continues to be on the rise. However, bacterial resistance to phage is another looming challenge capable of undermining the effectiveness of phage therapy. Moreover, the consideration of including phage therapy in modern medicine calls for more careful research around every aspect of phage study. In an attempt to adequately prepare for the events of phage resistance, many studies have attempted to experimentally evolve phage resistance in different bacterial strains, as well as train phages to evolve counter-infectivity of resistant bacterial mutants, in view of answering such questions as coevolutionary dynamics between phage and bacteria, mechanisms of phage resistance, fitness costs of phage resistance on bacteria, etc. In this review, we summarised many such studies and by careful examination, highlighted critical issues to the outcome of phage therapy. We also discuss the insufficiency of many of these in vitro studies to represent actual disease conditions during phage application, alongside other complications that exist in phage-bacterial evolutionary interactions. Conclusively, we present the exploitation of phage-bacterial interactions for successful infection managements, as well as some future perspectives to direct phage research.

LysP53 activity against Salmonella and its application in decontamination of Salmonella on fresh romaine lettuce.

Salmonella is a zoonotic pathogen that is commonly associated with foodborne disease outbreaks. This study found that a newly identified Gram-negative lysin LysP53 had good activity against a wide range of Salmonella, including Salmonella Newington, Salmonella Typhimurium, and Salmonella Dublin. Without the help of an outer membrane permeabilizer, 4 µM LysP53 could reduce 97.6% of planktonic Salmonella Enteritidis and 90% of the bacteria in biofilms. Moreover, LysP53 was highly thermostable because it maintained >90% activity even after exposure to temperatures up to 95 °C. Although high concentrations of salts could reduce the activity, LysP53 was found safe for oral gavage of mice without affecting body weights and cytokines in sera and able to reduce 90% of Salmonella Enteritidis loads on fresh romaine lettuce after 30 min of treatment. Because of its good activity against a wide range of bacteria, thermal stability, safe for oral administration, LysP53 could be used as a biocontrol agent for reducing bacterial loads in fresh vegetable food. KEY POINTS: • Lysin LysP53 has high bactericidal activity against Salmonella. • LysP53 is thermostable even at high temperature of up to 95 °C. • LysP53 can be used for topical decontamination of Salmonella on vegetables.

High-Resolution Ultrasound Imaging of Axillary Nerve and Relevant Injury.

OBJECTIVE: To evaluate the feasibility of axillary nerve (AN) visualization in healthy volunteers and the diagnostic value of AN injury via high-resolution ultrasonography (HRUS). METHODS: AN was examined by HRUS on both sides of 48 healthy volunteers and oriented the transducer according to three anatomical landmarks: quadrilateral space, anterior to subscapular muscle, and posterior to axillary artery. The maximum short-axis diameter (SD) and cross-sectional area (CSA) of AN were measured at different levels, and AN visibility was graded by using a five-point scale. The patients suspected of having AN injury were assessed by HRUS, and the HRUS features of AN injury were observed. RESULTS: AN can be visualized on both sides in all volunteers. There was no significant difference in SD and CSA of AN at the three levels between the left and right sides or in SD between males and females. However, the CSA of males at different levels was slightly larger than those of females (P < .05). In most volunteers, AN visibility at different levels was excellent or good, and AN was best displayed anterior to subscapular muscle. Rank correlation analysis revealed that the degree of AN visibility had correlation with height, weight, and BMI. A total of 15 patients diagnosed with AN injury, 12 patients showed diffuse swelling or focal thickening in AN, and 3 patients showed AN discontinuity. CONCLUSION: HRUS is able to reliably visualize AN, and it could be considered as the first choice for diagnosing AN injury.

Genetic Signatures from Adaptation of Bacteria to Lytic Phage Identify Potential Agents To Aid Phage Killing of Multidrug-Resistant Acinetobacter baumannii.

With the increasing morbidity and mortality rates associated with multidrug-resistant bacteria, interest in bacteriophage therapy has been revived. However, bacterial resistance to phage infection threatens the usefulness of phage therapy, especially its inclusion in modern medicine. Multidrug-resistant Acinetobacter baumannii is a top-priority pathogen requiring urgent intervention and new therapeutic approaches, such as phage therapy. Here, we experimentally adapted A. baumannii WHG40004 to its lytic phage P21 and thereafter isolated a phage-resistant bacterial mutant, named Ev5-WHG. We then aimed to identify potential agents to aid phage killing of Ev5-WHG by analyzing its genome and that of the wild-type strain. The enriched Gene Ontology (GO) analysis based on genetic alterations in minor alleles and mutations showed that pathways such as zinc ion transport and cell membrane synthesis could play certain roles in phage resistance. Remarkably, the combination of zinc acetate and P21 showed increased bactericidal effect on Ev5-WHG. Significantly also, we showed that P21 completely prevented the growth of wild-type WHG40004 in the presence of antibiotics (meropenem and imipenem). The results from this study indicate that the analysis of phage resistance signatures during adaptation of bacteria to a lytic phage can inform the choice of agents to work cooperatively with phage to limit and/or reverse resistance. This approach could be important for guiding future successful phage therapy. IMPORTANCE Bacteriophages have proven very useful as alternative therapeutic agents in combating multidrug-resistant bacterial infections; however, bacterial resistance to phages threatens their use. In this study, we showed a new strategy of leveraging genetic signatures that accompany phage resistance in bacteria to predict agents that can be used with lytic phages to combat multidrug-resistant Acinetobacter baumannii. Significantly, this approach was helpful in suggesting the use of zinc acetate to reduce resistance in phage-resistant bacteria, as well as the use of phage with antibiotics meropenem and imipenem to prevent resistance in a wild-type strain of multidrug-resistant A. baumannii. The approach of this study will be helpful for improving the outcome of phage therapy and in overcoming antimicrobial resistance.

The Long Noncoding RNA RP11-728F11.4 Promotes Atherosclerosis.

OBJECTIVE: Noncoding RNAs are emerging as important players in gene regulation and cardiovascular diseases. Their roles in the pathogenesis of atherosclerosis are not fully understood. The purpose of this study was to determine the role played by a previously uncharacterized long noncoding RNA, RP11-728F11.4, in the development of atherosclerosis and the mechanisms by which it acts. Approach and Results: Expression microarray analysis revealed that atherosclerotic plaques had increased expression of RP11-728F11.4 as well as the cognate gene FXYD6 (FXYD domain containing ion transport regulator 6), which encodes a modulator of Na+/K+-ATPase. In vitro experiments showed that RP11-728F11.4 interacted with the RNA-binding protein EWSR1 (Ewings sarcoma RNA binding protein-1) and upregulated FXYD6 expression. Lentivirus-induced overexpression of RP11-728F11.4 in cultured monocytes-derived macrophages resulted in higher Na+/K+-ATPase activity, intracellular cholesterol accumulation, and increased proinflammatory cytokine production. The effects of RP11-728F11.4 were enhanced by siRNA-mediated knockdown of EWSR1 and reduced by downregulation of FXYD domain containing ion transport regulator 6. In vivo experiments in apoE knockout mice fed a Western diet demonstrated that RP11-728F11.4 increased proinflammatory cytokine production and augmented atherosclerotic lesions. CONCLUSIONS: RP11-728F11.4 promotes atherosclerosis, with an influence on cholesterol homeostasis and proinflammatory molecule production, thus representing a potential therapeutic target. Graphic Abstract: A graphic abstract is available for this article.

Double contrast-enhanced ultrasound for the preoperative gross classification of gastric cancer: a comparison with multidetector computed tomography.

PURPOSE: To compare the diagnostic performance of double contrast-enhanced ultrasound (DCEUS) and multi-detector row computed tomography (MDCT) in the gross classification of gastric cancer (GC) preoperatively. METHODS: 54 patients with histology proved GC were included in this retrospective study. The sensitivity and specificity of DCEUS and MDCT for the gross classification of GC was calculated and compared. The area under the curve (AUC) from a receiver operating characteristic curve analysis was used to evaluate the difference of the diagnostic performance between these two methods. RESULTS: There were no significant differences between DCEUS and MDCT in terms of AUC for early gastric cancer (EGC), Borrmann I, II, III and Borrmann (III + IV) (P = 0.248, 0.317, 0.717, 0.464 and 0.594, respectively). The accuracy of DCEUS in diagnosing EGC, Borrmann I, II and Borrmann (III + IV) was higher than that of MDCT (96% vs 92%; 96% vs 94%; 87% vs 80%; 83% vs 73%), while in determining Borrmann III and IV, that of DCEUS was lower than that of MDCT (72% vs 74%; 89% vs 96%). CONCLUSION: Considering the revolution in clinical decision, prognosis evaluation, safety and non-invasion aspects, DCEUS can be used as the main alternative method for Borrmann classification of GC preoperatively.

Correlation between the spatial distribution and colony size was common for monogenetic bacteria in laboratory conditions.

BACKGROUND: Geographically separated population growth of microbes is a common phenomenon in microbial ecology. Colonies are representative of the morphological characteristics of this structured population growth. Pattern formation by single colonies has been intensively studied, whereas the spatial distribution of colonies is poorly investigated. RESULTS: The present study describes a first trial to address the questions of whether and how the spatial distribution of colonies determines the final colony size using the model microorganism Escherichia coli, colonies of which can be grown under well-controlled laboratory conditions. A computational tool for image processing was developed to evaluate colony density, colony size and size variation, and the Voronoi diagram was applied for spatial analysis of colonies with identical space resources. A positive correlation between the final colony size and the Voronoi area was commonly identified, independent of genomic and nutritional differences, which disturbed the colony size and size variation. CONCLUSIONS: This novel finding of a universal correlation between the spatial distribution and colony size not only indicated the fair distribution of spatial resources for monogenetic colonies growing with identical space resources but also indicated that the initial localization of the microbial colonies decided by chance determined the fate of the subsequent population growth. This study provides a valuable example for quantitative analysis of the complex microbial ecosystems by means of experimental ecology.

Lung Ultrasound Findings in Patients With Coronavirus Disease (COVID-19).

OBJECTIVE. Although chest CT is the standard imaging modality in early diagnosis and management of coronavirus disease (COVID-19), the use of lung ultrasound (US) presents some advantages over the use of chest CT and may play a complementary role in the workup of COVID-19. The objective of our study was to investigate US findings in patients with COVID-19 and the relationship of the US findings with the duration of symptoms and disease severity. MATERIALS AND METHODS. From March 3, 2020, to March 30, 2020, consecutive patients with a positive reverse transcriptase polymerase chain reaction test result for the virus that causes COVID-19 were enrolled in this study. Lung US was performed, and the imaging features were analyzed. The Fisher exact test was used to compare the percentages of patients with each US finding between groups with different symptom durations and disease severity. RESULTS. Our study population comprised 28 patients (14 men and 14 women; mean age ± SD, 59.8 ± 18.3 years; age range, 21-92 years). All 28 patients (100.0%, 28/28) had positive lung US findings. The most common findings were the following: B-lines (100.0%, 28/28), consolidation (67.9%, 19/28), and a thickened pleural line (60.7%, 17/28). A thickened pleural line was observed in a higher percentage of patients with a longer duration of the disease than in those with a shorter duration of the disease, and pulmonary consolidations were more common in severe and critical cases than in moderate cases. CONCLUSION. Typical lung US findings in patients with COVID-19 included B-lines, pulmonary consolidation, and a thickened pleural line. In addition, our results indicate that lung US findings can be be used to reflect both the infection duration and disease severity.

Contribution of genetic polymorphism of methylene tetrahydrofolate reductase on the effect of methotrexate in ectopic pregnancy patients.

BACKGROUND: Methotrexate (MTX) is the prior drug in ectopic pregnancy (EP). However, approximately 10% of patients suffer from failure by MTX therapy. Reduced folate carrier 1 (RFC1), methylene tetrahydrofolate reductase (MTHFR), and dihydrofolate reductase (DHFR) are involved in the transport and effects of MTX in vivo. In the present study, we aim to investigate the relationship between the genetic polymorphisms of RFC1, MTHFR, and DHFR and the clinical efficacy of MTX in tubal pregnancies. METHODS: 100 patients of EP were enrolled in this study. Polymorphisms of RFC1 G80A, MTHFR C677T, and DHFR A-317G were genotyped. ß-hCG level was detected in day 0, 4, and 7 after MTX injection. Association of MTX efficacy and genetic polymorphisms was analyzed. RESULTS: Methylene tetrahydrofolate reductase C677T was associated with MTX treatment (P = .017). The success rate of first MTX injection was superior in patients with harboring mutation allele of MTHFR gene than that in patients with wild-type gene (P = .001). However, there was no significant association between the polymorphisms of RFC1 G80A, DHFR A-317G, and surgical treatment (P = .709 and .476, respectively). In addition, ß-hCG level decrement was not significantly changed by MTX injection with different polymorphisms of RFC1, MTHFR, and DHFR on either day 4 (P = .214, 0.197 and 0.270, respectively) or day 7 (P = .172, .554, and .726, respectively). CONCLUSION: Our results suggested that the reliable indicator was polymorphism of MTHFR C677T in failure by MTX injection.

Long non-coding RNA LINC00858 aggravates the oncogenic phenotypes of ovarian cancer cells through miR-134-5p/RAD18 signaling.

PURPOSE: Ovarian cancer is a common gynecological cancer. Herein, we focused on the function and probable mechanisms of LINC00858 in ovarian cancer. METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR) was employed for detecting the expression of LINC00858, miR-134-5p and RAD18 E3 ubiquitin protein ligase (RAD18). Cell proliferation, migration, invasion, epithelial-mesenchymal transition (EMT) and apoptosis were detected by cell counting kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), transwell, terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) and western bolt experiments, as appropriate. Interplays between LINC00858, miR-134-5p and RAD18 were detected by RNA immunoprecipitation (RIP), RNA pull down and luciferase reporter assays. RESULTS: LINC00858 were up-regulated in ovarian cancer tissues and cells, and its expression was elevated in advanced samples compared to early ones. Knocking down LINC00858 inhibited cell proliferation, motility and EMT, but accelerated cell apoptosis in ovarian cancer. Moreover, could be sponged by LINC00858 sponged miR-134-5p to enhance RAD18 expression in ovarian cancer. Also, silenced RAD18 could also restrain oncogenic behaviors of ovarian cancer cells. Rescue experiments showed that overexpressing RAD18 reversed the effects caused by knocking down LINC00858 on cellular processes. CONCLUSION: LINC00858 sequestered miR-134-5p to elevate RAD18 expression, resulting in aggravated development of ovarian cancer. This might provide promising targets for treating patients with ovarian cancer.

Microarray profiling analysis and validation of novel long noncoding RNAs and mRNAs as potential biomarkers and their functions in atherosclerosis.

Long noncoding (lnc)RNAs have been implicated in the development and progression of atherosclerosis. However, the expression and mechanism of action of lncRNAs in atherosclerosis are still unclear. We implemented microarray analysis in human advanced atherosclerotic plaques and normal arterial intimae to detect the lncRNA and mRNA expression profile. Gene Ontology functional enrichment and pathway analyses were applied to explore the potential functions and pathways involved in the pathogenesis of atherosclerosis. A total of 236 lncRNAs and 488 mRNAs were selected for further Ingenuity Pathway Analysis. Moreover, quantitative RT-PCR tests of most selected lncRNAs and mRNAs with high fold changes were consistent with the microarray data. We also performed ELISA to investigate the corresponding proteins levels of selected genes and showed that serum levels of SPP1, CD36, ATP6V0D2, CHI3L1, MYH11, and BDNF were differentially expressed in patients with coronary heart disease compared with healthy subjects. These proteins correlated with some biochemical parameters used in the diagnosis of cardiovascular diseases. Furthermore, receiver operating characteristic analysis showed a favorable diagnostic performance. The microarray profiling analysis and validation of differentially-expressed lncRNAs and mRNAs in atherosclerosis not only provide new insights into the pathogenesis of this disease but may also reveal new biomarkers for its diagnosis and treatment.

Can Contrast-Enhanced Ultrasound Increase or Predict the Success Rate of Testicular Sperm Aspiration in Patients With Azoospermia?

OBJECTIVE. The objective of our study was to determine whether contrast-enhanced ultrasound (CEUS) perfusion measurements obtained before testicular sperm aspiration (TESA) can improve or predict sperm retrieval (SR) outcomes of TESA in patients with azoospermia. SUBJECTS AND METHODS. Between May 2017 and January 2018, 70 patients with azoospermia (mean age, 29 years; age range, 22-41 years) underwent testes CEUS within 10 days before TESA. Major perfusion areas were visually chosen, and their ranges were recorded. The other areas were defined as minor perfusion. CEUS quantitative features were acquired for both the main perfusion area and whole testis. Testis tissue biopsies were taken for both major and minor perfusion areas by cognitive fusion, and SR outcomes were compared. Associations between testicular volume, quantitative CEUS features, and SR outcomes were analyzed. RESULTS. Twenty-four men were found to have obstructive azoospermia (OA), and the remaining 46 had nonobstructive azoospermia (NOA). All patients with OA had spermatozoa in biopsy. Only one patient with NOA had spermatozoa in the major perfusion area but not the minor perfusion area; the other patients with NOA had the same SR outcomes in both major and minor perfusion areas. In patients with NOA, both wash-in and washout CEUS features were correlated with the success of SR in TESA. CONCLUSION. CEUS-guided TESA with cognitive fusion cannot yield improved SR outcomes of TESA in patients with NOA, possibly because of imprecise correlation between biopsy sites and main perfusion area analyzed by CEUS; however, quantitative CEUS features can be useful predictors of the success of SR.

Preoperative Tumor Staging of Gastric Cancer: Comparison of Double Contrast-Enhanced Ultrasound and Multidetector Computed Tomography.

OBJECTIVES: The purpose of this study was to compare the sensitivity and specificity of double contrast-enhanced ultrasound (CEUS) and multidetector computed tomography (MDCT) in the preoperative tumor staging of gastric cancer (GC) to stratify patients for suitable treatment. METHODS: Fifty-four patients with GC proved by histologic findings were included. The sensitivity and specificity of double CEUS and MDCT for tumor staging were calculated and compared. The differences between these methods were evaluated by using the area under the curve (AUC) from a receiver operating characteristic curve analysis. RESULTS: There were no significant differences in AUC values for T1 and T2 stages between double CEUS and MDCT (P = .190 and .256, respectively). However, the sensitivity of double CEUS in the detection of the T1 stage was higher than that of MDCT (88% versus 75%). The AUC values of MDCT for T3 and T4 stages were 0.833 and 0.905, which were both significantly higher than those of double CEUS (0.759 and 0.696; P < .05). The sensitivities of double CEUS and MDCT for the T3 stage were both 89%, but the accuracy and specificity of double CEUS were lower than those of MDCT (76% versus 83% and 63% versus 78%). The specificities of double CEUS and MDCT for the T4 stage were both 98%, but the accuracy and sensitivity of double CEUS were lower than those of MDCT (85% versus 94% and 42% versus 83%). CONCLUSIONS: Multidetector CT is superior to double CEUS for T3 and T4 GC, and double CEUS may be regarded as an important complementary method to MDCT.

[Values of Longitudinal and Transverse Plane Ratios in Predicting the Risk of Malignant Thyroid Nodules].

Objective To compare the predictive value of different ratio measurement in predicting the risk of malignant thyroid nodules and to determine the best value of ratio in diagnosing thyroid nodules.Methods The clinical data of 342 thyroid nodules diagnosed by ultrasonography and confirmed by histology in our hospital from January 2018 to August 2018 were analyzed.The ratio of nodules in different sections,including longitudinal plane ratio(A/TL)and transverse plane ratio(A/TC),was obtained through the maximum head-foot diameter(TL),the maximum left-right diameter(TC),and the anterior-posterior diameter(A)of transverse section measured by ultrasonography.The correlation of histological diagnosis of benign or malignant nodule with longitudinal ratio and transverse ratio were analyzed.Results The A/TC and A/TL of malignant nodules were 1.00(0.83,1.17)and 0.81(0.65,1.00),respectively,which were significantly higher than those of benign nodules [0.81(0.67,0.93)(Z=-6.567,P=0.000)and 0.63(0.52,0.75)(Z=-7.239,P=0.000)].The area under the ROC curve of A/TC and A/TL was 0.734 and 0.712,respectively,showing no significant difference(area difference:0.0210,standard error:0.0213,95% CI:-0.0207-0.0627,Z=0.987,P=0.3235).The threshold values of A/TC and A/TL for predicting malignant nodules were 0.784 161 5 and 0.985 714 5,respectively.Conclusions Both A/TL and A/TC has similar diagnostic value in predicting the risk of malignant nodules.The best cutoff value of the above two ratios are 0.78 and 0.99 respectively.

[Value of Elasticity Contrast Index of Ultrasonography in Differentiating Benign and Malignant Cervical Lymph Nodes].

Objective To investigate the diagnostic value of elasticity contrast index(ECI)in the differential diagnosis between benign and malignant cervical lymph nodes.Methods Cervical lymph nodes of 48 patients were examined by traditional ultrasound and ECI in Peking University Third Hospital between December 2016 and April 2017. Patients were further divided into free-hand group and non-free-hand group according to the use of manual compression or not. With pathological or clinical follow-up results as the gold standard,we compared the diagnostic values of traditional ultrasound,ECI,and their combination in the differential diagnosis between benign and malignant lymph nodes.Results Totally 138 lymph nodes were collected from 46 patients,among which 76 were benign and 62 were malignant. The ECI value of malignant lymph nodes(1.69±0.89)were significantly higher than that of benign lymph nodes(1.32±0.87)(t=-2.46,P=0.015).The area under receiver operating characteristic(ROC)curve for traditional ultrasound in discrimination of malignant cervical lymph nodes with benign ones was 0.899,with an optimal cut-off value of 7,which had a sensitivity of 93.5% and a specificity of 84.2%. The area under ROC curve for ECI was 0.649,with an optimal cut-off value of 1.25,which had a sensitivity of 59.7% and a specificity of 60.5%. For their combination,the area under ROC curve was 0.724,with a sensitivity of 93.5% and a specificity of 51.3%. In the group without using free-hand technique,the mean ECI value was 1.77±0.94 for malignant nodes,which was significantly higher than that(1.26±0.81)for benign nodes(t=-3.09,P=0.003). In the group requiring free-hand technique,the ECI value for malignant nodes was 1.33(1.01,1.44),showing no significant difference with benign ones[1.32(0.78,2.18);z=-0.20,P=0.843].Conclusions Conventional ultrasound has higher diagnostic value than ECI and their combination in differentiating benign and malignant lymph nodes. The role of ECI in evaluating other organs requires further investigations.

A transmembrane C-type lectin receptor mediates LECT2 effects on head kidney-derived monocytes/macrophages in a teleost, Plecoglossus altivelis.

Leukocyte cell-derived chemotaxin 2 (LECT2) is a multifunctional cytokine involved in many diseases in which immune dysfunction is present. Ayu LECT2 (PaLECT2), which interacts with a C-type lectin receptor (PaCLR), was shown to activate ayu head kidney-derived monocytes/macrophages (MO/MΦ) to improve the outcomes of fish upon bacterial infections. However, it is not known if PaCLR mediates PaLECT2 effects on ayu MO/MΦ. In this study, we determined the role of PaCLR in signal transduction of PaLECT2 on ayu MO/MΦ. We expressed the PaCLR ectodomain in Escherichia coli and produced a refolded recombinant protein (rPaCLR) that was then used to produce the anti-PaCLR IgG (anti-PaCLR) for neutralization. Addition of the refolded PaLECT2 mature peptide (rPaLECT2m) to ayu MO/MΦ cultures, increased cytokine expression, induced chemotaxis, and enhanced phagocytosis and bactericidal activity of these cells were observed. When we added anti-PaCLR to block the ectodomain of PaCLR, these effects were significantly inhibited. Based on our previous works and the data presented here, we conclude that PaCLR mediates the immunomodulatory effects of PaLECT2 on ayu MO/MΦ, thus defining a mechanism by which LECT2 protects fish against pathogens.

TRPM7 is required for ovarian cancer cell growth, migration and invasion.

Our previous study demonstrated that the melastatin-related transient receptor potential channel 7 (TRPM7) was highly expressed in ovarian carcinomas and its overexpression was significantly associated with poor prognosis in ovarian cancer patients. However, the function of TRPM7 in ovarian cancer is mostly unknown. In this study, we examined the roles of TRPM7 in ovarian cancer cell proliferation, migration and invasion. We found that short hairpin RNA interference-mediated silence of TRPM7 significantly inhibited cell proliferation, colony formation, migration and invasion in multiple ovarian cancer cell lines. Mechanistic investigation revealed that silence of TRPM7 decreased phosphorylation levels of Akt, Src and p38 and increased filamentous actin and focal adhesion number in ovarian cancer cells. Thus, our results suggest that TRPM7 is required for proliferation, migration and invasion of ovarian cancer cells through regulating multiple signaling transduction pathways and the formation of focal adhesions.