First Report of Paramyrothecium breviseta Causing Target Spot on Amorphophallus muelleri in Yunnan, China.

Amorphophallus muelleri is an Araceae plant with perennial tuber, widely used in food, pharmaceutical and chemical industry due to its richness in glucomannan. In April 2022, an outbreak of a target spot on A. muelleri plantlets was observed in a nursery in Ruili, Yunnan, China. The leafstalks of the diseased plantlets in the nursery turned brown and decayed (Fig.1 A-B), then gradually some water-soaked spots on the true leaves developed along the veins (Fig.1 A). Subquencely, the spots on the true leaves turned dark green to white-grayish in the center, which formed light to dark brown concentric rings with a target-like appearance surrounded by a yellow halo (Fig.1 C). When the temperature was 20-34℃ and the relatively humidity was 25-80%, dark-green to black sporodochia with white hypha appeared on the lower and upper leaf surfaces. Finally, 5-8% of the plants surveyed on 800 m2 of one-year-old plantlets in the nursery showed the symptoms and some plants with infected leafstalks would be death. Similar symptoms were also observed on about 10% of the transplanted plants surveyed on 12000 m2 (1.2 ha) of two-year-old plantlets in the field. Five diseased leaves from five distinct plantlets in the nursery were collected for pathogen isolation. Leaf pieces(5 x 5 mm) were cut from the edge of necrotic lesions, and surface-sterilized with 2.5% sodium hypochlorite for 1 min, 75% ethanol for 30 s, then rinsed 5 times by sterilized distilled water, finally put the leaf pieces on sterilized filter paper for 3-5 minutes to dry them and transferred onto potato dextrose agar (PDA) in petri dishes at 25℃ for three days. Five pure cultures identical to colony and conidial characteristics were isolated from five individual plants. The representative pure culture (M1) was grayish-white and circular colonies were 7.50 cm in diamter after 15 days at 25℃, with dark green concentric rings of sporodochia, the dorsal view of the colonies were yellowish. Conidia were aseptate, smooth, cylindrical, 5.00-6.25 (5.71) x 1.25-1.67 (1.63) µm (n = 20) rounded at both ends. A spore suspension (1 x 106 spores/ml) was prepared by harvesting spores from 15-day-old cultures grown in the dark at 25℃, then a thirty-ml of spore suspension was sprayed on the healthy leaves of 10 two-year-old plantlets. Thirty-ml of sterile water was sprayed on the healthy leaves of another 10 seedlings and used as the control. All seedlings were placed in a nursery at 20 to 34℃ and a relative humidity of 25 to 80%. Similar symptoms (Fig.1 D-F) to those observed in the nursery and field developed on all the 10 seedlings inoculated with M1 after two days, but not on the control leaves. The pathogenicity tests were repeated for three times. Fungal cultures reisolated from the infected leaves were identical to the original colonies and conidia, completing Koch's postulates. The internal transcribed spacer (ITS, primers ITS1 and ITS4) region of ribosomal DNA (OQ553785), calmodulin (cmdA, primers CAL-228F and CAL2Rd)(OQ559103), RNA polymerase II second largest subunit (rpb2, primers RPB2-5F2 and RPB2-7cR) (OQ559104) and ß-tubulin (tub2, primers Bt2a and Bt2b) (OQ559105) of M1 had 100%, 98.52%, 98.98% and 98.98% identity with the sequences of Paramyrothecium breviseta CBS544.75 (KU846289 for ITS, KU846262 for cmdA, KU846351 for rpb2, and KU846406 for tub2), respectively. In the phylogenic tree based on ITS, cmdA, rpb2 and tub2 gene sequences, the pure culture M1 clustered with P. breviseta CBS544.75, SDBR-CMU387, DRL4 and DRL3, which has been reported as the pathogen of leaf spot of Coffea arabica in China, C. canephora in China and Thailand (Wu et al. 2021; Withee et al. 2022). Molecular and morphological observations showed the pure culture M1 were P. breviseta (Withee et al. 2022), in addition the disease was named as target spot dueing to the typical target symptom on the leaves. To our knowledge, this is the first report of P. breviseta on A. muelleri from Yunnan, China, as well as worldwide. This disease can caused serious economic losses of A. muelleri dueing to that it can result 5-8% death of the plants in the nursery.

Evaluation of the Time Difference Method in Identifying Hepatocellular Carcinoma in Current CEUS LR-M Category Nodules.

OBJECTIVE: The aim of the work described here was to explore a potential method for improving the diagnostic detection of hepatocellular carcinoma (HCC) based on the contrast-enhanced ultrasound (CEUS) Liver Imaging Reporting and Data System (LI-RADS) Version 2017. METHODS: We retrospectively evaluated 585 liver nodules in 427 patients at risk for HCC from December 2020 to March 2023. The nodules were categorized as LR-1 to LR-M based on CEUS LI-RADS Version 2017 and were randomly subclassified into a developmental cohort (DC) and a validation cohort (VC) at 3:1. In the DC, the cutoff value of the time difference (∆T) for differentiating HCC from other malignancies by LR-M was calculated and used to reclassify nodules in the VC. The diagnostic effect on HCC detection before and after reclassification was further assessed. RESULTS: According to the current CEUS LI-RADS, 140 of 426 (32.9%) confirmed HCC nodules were misclassified as LR-M. In the DC (439 nodules), the receiver operating characteristic (ROC) curve revealed that the cutoff value of ∆T (wash-out onset time minus contrast arrival time) recommended for HCC diagnosis was greater than 21 s. In the VC (146 nodules), 34 HCCs were correctly categorized as LR-5 according to the cutoff value, and after reclassification, LR-5 had higher accuracy (67.1% vs. 89.0%, p < 0.001) and sensitivity (56.0% vs. 87.2%, p < 0.001) for HCC diagnosis with high specificity (100% vs. 94.6%, p = 0.500). CONCLUSION: Using the time difference method could identify HCC nodules misdiagnosed as LR-M and improve the diagnostic performance of current CEUS LI-RADS.

Isoalantolactone mediates the degradation of BCR-ABL protein in imatinib-resistant CML cells by down-regulating survivin.

Isoalantolactone (Iso) is a bioactive lactone isolated from the root of Inula helenium L, which has been reported to have many pharmacological effects. To investigate the role and mechanism of isoalantolactone in chronic myeloid leukemia (CML), we first investigated isoalantolactone's anti-proliferative effects on imatinib-sensitive and imatinib-resistant CML cells by CCK8. Flow cytometry was used to detect isoalantolactone-induced cell apoptosis. Survivin was overexpressed in KBM5 and KBM5T315I cells using the lentivirus vector pSIN-3×flag-PURO. In KBM5 and KBM5T315I cells, shRNA was used to knockdown survivin. Cellular Thermal Shift Assay (CETSA) was used to detect the interaction between isoalantolactone and survivin. The ubiquitin of survivin induced by isoalantolactone was detected through immunoprecipitation. Quantitative polymerase-chain reaction (Q-PCR) and western blotting were used to detect the levels of mRNA and protein. Isoalantolactone inhibits the proliferation and promotes apoptosis of imatinib-resistant CML cells. Although isoalantolactone inhibits the proteins of BCR-ABL and survivin, it cannot inhibit survivin and BCR-ABL mRNA levels. Simultaneously, it was shown that isoalantolactone can degrade survivin protein by increasing ubiquitination. It was demonstrated that isoalantolactone-induced survivin mediated downregulation of BCR-ABL protein. It was also revealed that isoalantolactone triggered BCR-ABL protein degradation via caspase-3. Altogether, isoalantolactone inhibits survivin through the ubiquitin proteasome pathway, and mediates BCR-ABL downregulation in a caspase-3 dependent manner. These data suggest that isoalantolactone is a natural compound, which can be used as a potential drug to treat TKI-resistant CML.

The public health response to an outbreak of border-spill malaria along China-Myanmar border.

INTRODUCTION: Malaria importation can be caused by cross-border movement either of both people and anopheline mosquitoes. However, there still lacks robust evidence of imported malaria caused by Plasmodium spp. infected anopheles along international border areas (border-spill malaria). The objectives of this study were to confirm whether an outbreak of Plasmodium vivax malaria is border-spill malaria and assess the effects of China's public health response along China-Myanmar border. METHODS: Epidemiological, parasitological and entomological investigations were conducted to investigate the outbreak of border-spill malaria. Meanwhile, comprehensive interventions were carried out to prevent further transmission and reintroduction of malaria. RESULTS: Rapid diagnostic testing, microscopy and polymerase chain reaction were performed and the infections were confirmed as P. vivax. A total of 22 (9.21%) of 239 workers contracted P. vivax during the outbreak. Multivariate logistic regression analysis identified that the distance of worker shelters in China within 300 meters to the internally displaced person (IDP) camps in Myanmar was a risk factors associated with malaria infection (adjusted odds ratio 7.5920; 95% confidence interval, 2.6079-22.1013; P = 0.0002). After comprehensive interventions, malaria transmission was successfully interpreted and prevented at the project site till the completion of project on 14 January 2020, and recurrence of P. vivax malaria was not detected by the end of 2020. CONCLUSION: This study provided robust evidence of border-spill malaria along China-Myanmar border. Malaria parasite reservoir and distance travelled by female anopheline mosquitoes are two determinants for border-spill malaria. The public health response to the outbreak indicates that the malaria surveillance and response system works well in preventing reintroduction of malaria. However, prevention of border-spill malaria is still a major challenge in the Yunnan border area, China.

Diagnostic value of color parametric imaging and contrast-enhanced ultrasound in the differentiation of hepatocellular adenoma and well-differentiated hepatocellular carcinoma.

PURPOSE: To investigate the differential diagnostic value of computer-aided color parametric imaging (CPI) and contrast-enhanced ultrasound (CEUS) in hepatocellular adenoma (HCA) and well-differentiated hepatocellular carcinoma (wHCC). METHOD: A total of 38 patients who underwent CEUS and were pathologically diagnosed with HCA (10 cases) and wHCC (28 cases) were reviewed retrospectively. The differences between the radiological features of HCA and wHCC were compared by two readers, blinded to the final diagnosis. RESULTS: (a) Sonographic features: on gray-scale ultrasound, halo sign was more common in wHCC than in HCA (60.7% vs. 10.0%, p = 0.009). On CEUS, hyper- or isoenhancement was more common in HCA in the portal phase (90.0% vs. 50.0%; p = 0.022). On CPI mode, HCA was inclined toward centripetal enhancement (60.0% vs. 14.3% p = 0.010). HCA was characterized by the presence of pseudocapsule enhancement (50.0% vs. 14.3%; p = 0.036). Quantitative analysis showed that the arrival time of HCA was earlier than that of wHCC (12.4 ± 3.7 s vs. 15.9 ± 3.2 s; p = 0.006). (b) Interobserver agreement was improved by using CPI compared with CEUS. The diagnostic sensitivity, specificity, and accuracy of the combination were 80.0%, 85.7%, and 84.2%, respectively. CONCLUSIONS: CEUS combined with CPI can provide effective information for the differential diagnosis of HCA and wHCC, especially for the non-experienced radiologists.

Alkaline phosphatase-to-albumin ratio as a novel predictor of long-term adverse outcomes in coronary artery disease patients who underwent PCI.

BACKGROUND: Alkaline phosphatase (ALP) and albumin (ALB) have been shown to be associated with coronary artery disease (CAD), and it has been reported that alkaline phosphatase-to-albumin ratio (AAR) is associated with the liver damage and poorer prognosis of patients with digestive system malignancy. Moreover, several previous studies showed that there was a higher incidence of malignancy in CAD patients. However, to our knowledge, the relationship between AAR and long-term adverse outcomes in CAD patients after undergoing percutaneous coronary intervention (PCI) has not been investigated. Therefore, we aim to access the relation between AAR and long-term adverse outcomes in post-PCI patients with CAD. METHODS: A total of 3378 post-PCI patients with CAD were enrolled in the retrospective Clinical Outcomes and Risk Factors of Patients with Coronary Heart Disease after PCI (CORFCHD-ZZ) study from January 2013 to December 2017. The median duration of follow-up was 37.59 ± 22.24 months. The primary end point was long-term mortality including all-cause mortality (ACM) and cardiac mortality (CM). The secondary end points were major adverse cardiac events (MACEs) and major adverse cardiac and cerebrovascular events (MACCEs). RESULTS: Kaplan-Meier analyses showed that an increased AAR was positively correlated with incidences of long-term ACM (log-rank, P=0.014), CM (log-rank, P=0.011), MACEs (log-rank, P=0.013) and MACCEs (log-rank, P=0.006). Multivariate Cox regression analyses showed that the elevated AAR was an independent predictor of long-term ACM (adjusted HR = 1.488 [1.031-2.149], P=0.034), CM (adjusted HR = 1.837 [1.141-2.959], P=0.012), MACEs (adjusted HR = 1.257 [1.018-1.551], P=0.033) and MACCEs (adjusted HR = 1.237 [1.029-1.486], P=0.024). CONCLUSION: An elevated AAR is a novel independent predictor of long-term adverse outcomes in CAD patients following PCI.

Tumor microenvironment immune-related lncRNA signature for patients with melanoma.

BACKGROUND: The incidence of malignant melanoma accounts for only approximately 5% of skin malignant tumors, however, it accounts for 75% of its mortality. Long-chain non-coding RNA (lncRNA) has a wide range of functional activities. Disorders of lncRNAs may lead to the occurrence and development of melanoma, and may also be related to immunotherapy. METHODS: The transcriptomic data of primary and metastatic melanoma patients and 331 immune-related genes were downloaded from skin cutaneous melanoma (SKCM) in the The Cancer Genome Atlas (TCGA) database. On this basis, 460 immunologically relevant lncRNAs were identified by constructing a co-expression network of immunogenic genes and lncRNAs in primary and metastatic melanoma patients. Prognostic genes were screened using univariate Cox regression analysis. ROC analysis was performed to evaluate the robustness of the prognostic signature. RESULTS: Univariate correlation analysis showed that only 3 of the 23 immune-related lncRNAs were at high risk and the rest were at low risk. Signatures of 7 immune-related lncRNAs were identified by multivariate correlation analysis. The clinical correlation analysis showed that the 7 immune-related lncRNAs were associated with the clinical stage of primary and metastatic melanoma. Principal component analysis (PCA) showed that only 7 immune-related lncRNA signals divided tumor patients into high-risk and low-risk groups, while the low-risk group was enriched in the immune system process M13664 and immune response M19817 sets. PPI interaction network analysis showed that 11 G protein-coupled receptors and 6 corresponding ligands in the 2 gene sets affected the tumor microenvironment and were negatively related to the risk of the 7 immune-related lncRNAs. The tumor microenvironment immune cell infiltration analysis also supported the finding that anti-tumor immunity in the low-risk group was stronger than in the high-risk group. CONCLUSIONS: These results indicate that characteristics of the 7 immune-related lncRNAs have prognostic value for melanoma patients and can be used as potential immunotherapy targets.

Diversities of HLA-A, -B, -C, -DRB1 and -DQB1 loci in Chinese Kazak population and its genetic relatedness dissection with multiple populations: a comparative study.

Studying the allele and haplotype distributions of human leukocyte antigen (HLA) loci at 2nd-field level in different populations was important. Allele and haplotype frequencies of HLA-A, -B, -C, -DRB1 and -DQB1 loci in 110 unrelated healthy Kazak individuals living in Xinjiang (China) were analyzed using polymerase chain reaction sequence based typing. Thirty HLA-A, 48 HLA-B, 24 HLA-C, 34 HLA-DRB1 and 18 HLA-DQB1 alleles were detected at the 2nd-field level in the Kazak population. Frequencies of HLA alleles, genotypes, and haplotypes were calculated, and some exhibited significantly different distributions among different populations. A neighbor-joining (NJ) tree, heatmap, multidimensional scaling (MDS) and principal component analysis (PCA) were used to explore the genetic relationships between the Kazak population and 32 reference populations distributed in Asia, Africa, America and Europe using frequency data of HLA-A, -B, -C and -DRB1 loci. The NJ tree, heatmap, and MDS of the 33 populations were constructed based on pairwise DA values of populations obtained by the HLA-A, -B, -C and -DRB1 allele frequencies. Different PCA plots were constructed based on the allele frequencies of HLA-A, -B, -C and -DRB1 or estimated haplotypic frequencies of HLA-A, -B, -C loci. The data obtained in the present research can be used for research on HLA-related diseases or paternity relationships, and aid to finding the best matched donors in stem cell transplantation for Kazak individuals.

Contrast-Enhanced Intraoperative Ultrasound Improved Sensitivity and Positive Predictive Value in Colorectal Liver Metastasis: a Systematic Review and Meta-Analysis.

BACKGROUND: The diagnostic accuracy of imaging modalities in colorectal cancer liver metastases (CRLM) has improved in recent years, therefore the role of current imaging techniques needs to be defined. OBJECTIVE: The aim of this study was to assess the diagnostic performance of magnetic resonance imaging, preoperative imaging (magnetic resonance imaging or computed tomography), intraoperative ultrasound, and contrast-enhanced intraoperative ultrasound in the detection of CRLM. MATERIALS AND METHODS: Eligible trials published before 30 March 2020 were identified from the EMBASE, PubMed, Web of Science, and Cochrane Library databases, and descriptive and quantitative data were extracted. Study quality was evaluated for the identified studies and a random-effects model was used to determine the integrated diagnosis estimation. Meta-regression was implemented to explore the possible contributors to heterogeneity. RESULTS: Overall, 13 studies were included for analysis, comprising 682 patients with a total of 2303 liver lesions. The pooled sensitivity, specificity, and diagnostic odds ratio of contrast-enhanced intraoperative ultrasound were 0.94 (95% confidence interval [CI] 0.89-0.97), 0.83 (95% CI 0.67-0.92), and 79 (95% CI 32-196), respectively. The overall weighted area under the curve was 0.96 (95% CI 0.94-0.97). In univariate meta-regression analysis, disappearing liver metastasis, contrast agent, and Kupffer phase were the potent sources of heterogeneity; however, in multivariate meta-regression, no definite variable was the source of the study heterogeneity. CONCLUSION: Contrast-enhanced intraoperative ultrasound demonstrated a high sensitivity and specificity for screening CRLM.

Molecular Mechanism of Tumor Cell Immune Escape Mediated by CD24/Siglec-10.

Tumor immune escape is an important part of tumorigenesis and development. Tumor cells can develop a variety of immunosuppressive mechanisms to combat tumor immunity. Exploring tumor cells that escape immune surveillance through the molecular mechanism of related immunosuppression in-depth is helpful to develop the treatment strategies of targeted tumor immune escape. The latest studies show that CD24 on the surface of tumor cells interacts with Siglec-10 on the surface of immune cells to promote the immune escape of tumor cells. It is necessary to comment on the molecular mechanism of inhibiting the activation of immune cells through the interaction between CD24 on tumor cells and Siglec-10 on immune cells, and a treatment strategy of tumors through targeting CD24 on the surface of tumor cells or Siglec-10 on immune cells.

Simultaneous determination of legacy and emerging organophosphorus flame retardants and plasticizers in indoor dust using liquid and gas chromatography-tandem mass spectrometry: method development, validation, and application.

In the present study, an analytical method has been developed and validated for the simultaneous detection and quantification of 19 PFRs (14 legacy organophosphorus flame retardants (PFRs) and 5 emerging PFRs (ePFRs)) and 20 plasticizers (7 legacy plasticizers (LPs) and 13 alternative plasticizers (APs)). Sample preparation was based on the combination of previously validated analytical protocols including ultrasonic extraction and Florisil fractionation/cleanup. The analysis was performed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for all targeted compounds, except for bis (2-ethylhexyl) phthalate (DEHP) and bis (2-ethylhexyl) terephthalate (DEHT), for which the separation of the isomers resulted in more favorable gas chromatography-electron ionization-mass spectrometry (GC-EI-MS). The new method was in-house validated by applying two levels of fortification in dust. The achieved linearity (R2) ranged between 0.993 and 0.999. Limits of detection and quantification (LODs and LOQs) ranged between 1 and 265 ng/g and between 1 and 870 ng/g for all analytes, respectively, except for DEHP and DEHT, for which relatively higher LODs (665 and 1100 ng/g, respectively) and LOQs (2100 and 3500 ng/g, respectively) were observed. Accuracy ranged between 75 and 125% for most of the targeted analytes, and repeatability was good with relative standard deviation (RSD) < 15% for most compounds. Finally, the method was applied for the determination and quantification of the targeted chemicals in house dust samples (n = 10) from the megacity of Guangzhou (China). Median values ranged from 3 to 210 ng/g for PFRs, from 4 to 165 ng/g for ePFRs, from 30 to 100,000 ng/g for LPs, and from 6 to 34,000 ng/g for APs. Main contributors to the total contamination were LPs 63% and APs 37% in total plasticizers, whereas PFRs and ePFRs contributed 90% and 10% in total flame retardants. Graphical abstract.

Thermosensitive liposomal doxorubicin plus radiofrequency ablation increased tumor destruction and improved survival in patients with medium and large hepatocellular carcinoma: A randomized, double-blinded, dummy-controlled clinical trial in a single center.

BACKGROUND: Lyso-thermosensitive liposomal doxorubicin (LTLD, ThermoDox) consists of doxorubicin encapsulated contained within a heat-sensitive liposome. AIMS AND OBJECTIVES: We sought to evaluate whether the use of combined radiofrequency ablation (RFA) and LTLD would result in larger coagulation volume and longer overall survival (OS) compared with the use of RFA alone in patients with 3-7 cm unresectable hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Between 2010 and 2012, 22 HCC patients were randomly assigned to one of two treatments in our center: (1) ultrasound-guided percutaneous RFA plus intravenous (IV) infusion of LTLD (combination, n = 11) or (2) RFA plus IV dummy (RFA, n = 11). Four patients withdrew from the study, and the remaining 18 patients entered the final analysis. There were 14 male and 4 female patients with an average age of 61.1 ± 9.3 years (range: 40-73 years). The average tumor size was 4.2 ± 1.0 cm (range: 3.1-6.1 cm). One-month enhanced computed tomography was used to evaluate the ablation efficacy and coagulation volume after RFA. Regular follow-up after RFA was performed to assess toxicity, local response rates, and OS rates. RESULTS: A major complication (empyema) occurred in one case in the combination group. Combination treatment region did not induce any additional toxicity beyond doxorubicin. The primary ablation success rate was 93.3% (14/15 tumors) in the combination group and 77.8% (7/9 tumors) in the RFA group (P = 0.308). The difference in coagulation volume between pre- and post-RFA in the combination group was significantly larger than that of the RFA group (105.7 ± 73.8 cm 3 vs. 37.3 ± 8.5 cm 3, P = 0.013). The follow-up period ranged from 11 to 80 months (average: 49.1 ± 24.8 months). The local progression rate was 6.7% (1/15 tumors) in the combination group and 22.2% (2/9 tumors) in the RFA group. The mean OS for the combination group was 68.5 ± 7.2 months, which was significantly greater compared with the RFA group (46.0 ± 10.6 months, P = 0.045). CONCLUSIONS: RFA with heat target delivery chemotherapy facilitated better tumor coagulation necrosis without additional toxicity. This combined treatment may improve the clinical efficacy of RFA or free doxorubicin and prolong survival in patients with medium to large HCC.

Assessment of the major odor contributors and health risks of volatile compounds in three disposal technologies for municipal solid waste.

Gaseous emissions from municipal solid waste (MSW) disposal plants pose serious odor pollution and health risks. In this study, the emission of volatile organic compounds and carbon disulfide was compared in the main processing units of three disposal methods, i.e., landfilling, eco-mechanical biological treatment (EMBT) and anaerobic fermentation in a MSW disposal plant. Among the detected volatile compounds (VCs), the top ten odor compounds were methanethiol, dimethyl sulfide, dimethyl disulfide, carbon disulfide, styrene, m-xylene, 4-ethyltoluene, ethylbenzene, 2-hexyl ketone and n-hexane in the MSW disposal plant. Sulfur compounds were the main source of odor at the majority of sampling sites, and aromatic compounds were the dominant odor substrates at the tipping unit and sorting system of EMBT, while 2-hexanone was the major odor substrate at the tipping unit (AT) and sorting system (AS) of anaerobic fermentation and the landfill working surface. At AS and AT, the lifetime cancer risk values for 1,2-dichloroethane and trichloroethylene exceeded the carcinogenic risk value (>1.0E-04), and the hazard index values of naphthalene, trichloroethylene and acrolein all exceeded the acceptable level (>1). Therefore, special attention should be paid to VC emissions from MSW disposal facilities, and protection measures should be adopted for on-site workers to minimize health risks.

Bioconcentration and biotransformation of organophosphorus flame retardants (PFRs) in common carp (Cyprinus carpio).

Understanding the bioaccumulation and biotransformation of xenobiotic compounds is critical for evaluating their fate and potential toxicity in vivo. In the present study, the tissue specific accumulation and depuration of seven organophosphorus flame retardants (PFRs) in common carp (Cyprinus carpio) were investigated after exposing the fish to an environmental relevant level of PFRs. The log Kow of PFRs was significantly negatively correlated to the percentages of individual PFRs to the total PFRs in serum (p < 0.04), whereas significantly positive correlations were observed in all other tissues (p < 0.02). Significant correlations (p < 0.01) between the log Kow of PFRs and their log bioconcentration factor (BCFww) were also found in all investigated tissues except for serum. This suggests that the hydrophobicity of PFRs played a significant role in the distribution and body compartment accumulation of PFRs in common carp. The bioaccumulation potential of PFRs in serum was different from the other tissues, probably due to its specific properties. Dialkyl and/or diaryl phosphate esters (DAP) and hydroxylated PFRs (HO-PFRs) were quantified as the major metabolites. Their levels in liver and intestine were significantly higher than in other tissues. Biotransformation processes also played a crucial role in the accumulation of PFRs in fish. Our results provide critical information for further understanding the bioconcentration, tissue distribution and metabolism of PFRs in fish.

Stereoselective Oxidation Kinetics of Deoxycholate in Recombinant and Microsomal CYP3A Enzymes: Deoxycholate 19-Hydroxylation Is an In Vitro Marker of CYP3A7 Activity.

The primary bile acids (BAs) synthesized from cholesterol in the liver are converted to secondary BAs by gut microbiota. It was recently disclosed that the major secondary BA, deoxycholate (DCA) species, is stereoselectively oxidized to tertiary BAs exclusively by CYP3A enzymes. This work subsequently investigated the in vitro oxidation kinetics of DCA at C-1ß, C-3ß, C-4ß, C-5ß, C-6α, C-6ß, and C-19 in recombinant CYP3A enzymes and naive enzymes in human liver microsomes (HLMs). The stereoselective oxidation of DCA fit well with Hill kinetics at 1-300 µM in both recombinant CYP3A enzymes and pooled HLMs. With no contributions or trace contributions from CYP3A5, CYP3A7 favors oxidation at C-19, C-4ß, C-6α, C-3ß, and C-1ß, whereas CYP3A4 favors the oxidation at C-5ß and C-6ß compared with each other. Correlation between DCA oxidation and testosterone 6ß-hydroxylation in 14 adult single-donor HLMs provided proof-of-concept evidence that DCA 19-hydroxylation is an in vitro marker reaction for CYP3A7 activity, whereas oxidation at other sites represents mixed indicators for CYP3A4 and CYP3A7 activities. Deactivation caused by DCA-induced cytochrome P450-cytochrome P420 conversion, as shown by the spectral titrations of isolated CYP3A proteins, was observed when DCA levels were near or higher than the critical micelle concentration (about 1500 µM). Unlike CYP3A4, CYP3A7 showed abnormally elevated activities at 500 and 750 µM, which might be associated with an altered affinity for DCA multimers. The disclosed kinetic and functional roles of CYP3A isoforms in disposing of the gut bacteria-derived DCA may help in understanding the structural and functional mechanisms of CYP3A.

Continuum of Host-Gut Microbial Co-metabolism: Host CYP3A4/3A7 are Responsible for Tertiary Oxidations of Deoxycholate Species.

The gut microbiota modifies endogenous primary bile acids (BAs) to produce exogenous secondary BAs, which may be further metabolized by cytochrome P450 enzymes (P450s). Our primary aim was to examine how the host adapts to the stress of microbe-derived secondary BAs by P450-mediated oxidative modifications on the steroid nucleus. Five unconjugated tri-hydroxyl BAs that were structurally and/or biologically associated with deoxycholate (DCA) were determined in human biologic samples by liquid chromatography-tandem mass spectrometry in combination with enzyme-digestion techniques. They were identified as DCA-19-ol, DCA-6ß-ol, DCA-5ß-ol, DCA-6α-ol, DCA-1ß-ol, and DCA-4ß-ol based on matching in-laboratory synthesized standards. Metabolic inhibition assays in human liver microsomes and recombinant P450 assays revealed that CYP3A4 and CYP3A7 were responsible for the regioselective oxidations of both DCA and its conjugated forms, glycodeoxycholate (GDCA) and taurodeoxycholate (TDCA). The modification of secondary BAs to tertiary BAs defines a host liver (primary BAs)-gut microbiota (secondary BAs)-host liver (tertiary BAs) axis. The regioselective oxidations of DCA, GDCA, and TDCA by CYP3A4 and CYP3A7 may help eliminate host-toxic DCA species. The 19- and 4ß-hydroxylation of DCA species demonstrated outstanding CYP3A7 selectivity and may be useful as indicators of CYP3A7 activity.

The value of KRAS gene status in predicting local tumor progression of colorectal liver metastases following radiofrequency ablation.

PURPOSE: We investigated the relationships between KRAS gene status and local tumor progression (LTP) of colorectal liver metastases (CLMs) after treatment with percutaneous ultrasound-guided radiofrequency ablation (RFA). MATERIALS AND METHODS: Clinical and imaging data from 76 patients (154 lesions) with CLM who underwent percutaneous ultrasound-guided RFA and had KRAS gene test results between January 2012 and June 2016 were analyzed. The average lesion size was 2.3 ± 1.0 cm (range 0.9-5.7 cm); 38 cases (82 lesions) had wild-type KRAS, and 38 cases (72 lesions) had KRAS mutations. RESULTS: The technique effectiveness was 98.1% (151/154), and the LTP rate was 18.2% (28/154) after RFA, which was performed between January 2012 and November 2017. The mean and median follow-up were 32.7 ± 2.5 and 32.0 ± 2.6 months (range 1-70 months), respectively. Cumulative LTP rates at 6 months and 1, 2 and 3 years post-RFA for all patients were 7.4, 14.5, 17.8 and 19.2%, respectively. The LTP rate for patients with mutant KRAS (27.8% [20/72]) was significantly higher than that in patients with wild-type KRAS (9.8% [8/82]; p = .004). The cumulative LTP rates at 6 months and 1, 2 and 3 years post-RFA were 4.0, 11.1, 11.1 and 11.1%, respectively, for patients with wild-type KRAS and 11.2, 18.4, 25.2 and 36.2%, respectively, for patient with mutant KRAS (p = .011). Univariate (p = .011) and multivariate analyses (p = .005) showed that KRAS genotype in liver metastases was predictive of LTP. Multivariate analysis also showed that ablation margin size (p< .001) and modified clinical risk score (CRS; p = .033) were independent prognostic factors for LTP. CONCLUSIONS: KRAS gene status of liver metastatic lesions was associated with LTP rates after RFA of CLM. Ablation margin size and modified CRS were also independent prognostic factors for LTP.

Diagnostic Value of Arrival Time Parametric Imaging Using Contrast-Enhanced Ultrasonography in Superficial Enlarged Lymph Nodes.

OBJECTIVE: The aim of this study was to evaluate the value of arrival-time parametric imaging for differential diagnosis of superficial enlarged lymph nodes. METHODS: Patients with lymphadenopathy who received contrast-enhanced ultrasonography (CEUS) and biopsy were included in this study. Following CEUS, a prototype software of the arrival-time parametric imaging system was used to analyze the video footage. Arrival-time patterns during the arterial phase were evaluated. The quantitative parameters including arrival time of periphery, arrival time of center, and the travel time (△T) were calculated. RESULTS: A total of 145 lymph nodes were analyzed. Arrival-time parametric imaging showed that 80.3% of metastatic lymph nodes and 68.4% of lymphoid tuberculosis presented a centripetal perfusion pattern, 76.5% of lymphoma showed complete homogeneous enhancement, and 81.2% of reactive lymph nodes had centrifugal patterns. The arrival time of periphery (sec) of metastatic lymph nodes was substantially earlier than that of lymphoma (11.0 ± 3.1 versus 12.6 ± 3.6; P < .05). The arrival time of center (sec) of metastatic lymph nodes was obviously later than that of lymphoma and reactive lymph nodes (13.4 ± 3.3 versus 10.5 ± 2.9 and 10.6 ± 1.5; P < .05). The travel time (△T) (sec) in metastatic lymph nodes was substantially longer than in reactive lymph nodes and lymphoma (4.2 ± 2.1 versus 2.3 ± 1.6 and 2.9 ± 2.5; P < .05). At a △T cutoff value of 2.75 seconds (using the receiver operating characteristic curve), the sensitivity and specificity in differentiating metastatic lymph nodes from benign lymph nodes (lymphoid tuberculosis and reactive lymph nodes) were 78.9% and 64.7%, respectively. CONCLUSIONS: Enhanced patterns and parameters of arrival-time parametric imaging during CEUS could provide more information for the differential diagnosis of enlarged superficial lymph nodes.

Diagnostic Value of Contrast-Enhanced Ultrasonography and Time-Intensity Curve in Differential Diagnosis of Cervical Metastatic and Tuberculous Lymph Nodes.

OBJECTIVES: To evaluate the diagnostic value of contrast-enhanced ultrasonography (CEUS) in the differential diagnosis of tuberculous and metastatic lymph nodes. METHODS: Nineteen cervical tuberculous lymph nodes and 43 cervical metastatic lymph nodes were investigated. The CEUS perfusion patterns and parameters of time-intensity curve (TIC) were analyzed. Diagnostic accuracy and consistency of two physicians were compared before and after CEUS and TIC analysis. RESULTS: Conventional ultrasonography showed no significant differences between tuberculous and metastatic lymph nodes. Concentric enhancement at the arterial phase of CEUS occurred in 15 of 19 (78.9%) tuberculous lymph nodes and 42 of 43 (97.7%) metastatic lymph nodes (P < .05). For the TIC curve, a steep descending curve with an apparent notch was commonly found in tuberculous lymph nodes (13 of 16). Although a shallow descending curve was common (40 of 43) in metastatic lymph nodes, most did not have a notch on the curve (39 of 43) (P < .01). The k-value and the peak intensity (PI) value of tuberculous lymph nodes were significantly higher and the △PI value was significantly lower than that of metastatic lymph nodes (P < .05, respectively). Kappa values for the diagnosis consistency of the two physicians before and after CEUS and TIC analysis were 0.582 and 0.761, respectively. Diagnostic accuracy before and after CEUS and TIC analysis was 47.4% (28 of 59) and 96.6% (57 of 59), respectively (P < .001). CONCLUSIONS: Contrast-enhanced ultrasonography with TIC analysis is helpful in differentiating tuberculous from metastatic lymph nodes.