[Component identification and analysis in vivo of Sanhan Huashi formula].

Sanhan Huashi formula(SHF) is the intermediate of a newly approved traditional Chinese medicine(TCM) Sanhan Huashi Granules for the treatment of COVID-19 infection. The chemical composition of SHF is complex since it contains 20 single herbal medicines. In this study, UHPLC-Orbitrap Exploris 240 was used to identify the chemical components in SHF and in rat plasma, lung and feces after oral administration of SHF, and heat map was plotted for characterizing the distribution of the chemical components. Chromatographic separation was conducted on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 µm) using 0.1% formic acid(A)-acetonitrile(B) as mobile phases in a gradient elution. Electrospray ionization(ESI) source was used to acquire data in positive and negative mode. By reference to quasi-molecular ions and MS/MS fragment ions and in combination with MS spectra of reference substances and compound information in literature reports, 80 components were identified in SHF, including 14 flavonoids, 13 coumarins, 5 lignans, 12 amino-compounds, 6 terpenes and 30 other compounds; 40 chemical components were identified in rat plasma, 27 in lung and 56 in feces. Component identification and characterization of SHF in vitro and in vivo lay foundations for disclosure of its pharmacodynamic substances and elucidation of the scientific connotation.

[Robot-assisted laparoscopic radical prostatectomy helps improve postoperative urinary continence recovery in patients with prostate cancer].

OBJECTIVE: To observe the recovery of urinary continence through postoperative rehabilitation training after robot-assisted laparoscopic radical prostatectomy (RARP) versus that after traditional laparoscopic radical prostatectomy (LRP). METHODS: This study included 64 cases of urinary incontinence after surgically treated for PCa from May 2017 to February 2021, 32 by RARP and the other 32 by LRP as the controls. All the patients received standard urinary continence rehabilitation training and routine nursing care postoperatively, followed by comparison of the rate and time of urinary incontinence recovery and the patients' scores on the quality of life (QOL) and satisfaction with treatment between the two groups. RESULTS: There were no statistically significant differences in age, PSA level or pathological stage between the two groups of patients (P > 0.05). After standard urinary continence rehabilitation training, the patients in the RARP group, compared with those in the LRP control, showed a lower grade of urinary incontinence (χ2 = 6.483, P = 0.039), a shorter mean duration of urinary incontinence per day, an earlier recovery of urinary continence (χ2 = 4.73, P = 0.030 at 1－3 months; χ2 = 12.696, P < 0.001 at 4－6 months), a higher rate of overall recovery (χ2 = 13.396, P = 0.004), and higher scores on QOL and satisfaction with treatment. CONCLUSION: RARP can effectively improve the recovery from postoperative urinary incontinence in PCa patients.

Lewis Acid Catalyzed [4 + 2] Cycloaddition of N-Tosylhydrazones with ortho-Quinone Methides.

A formal [4 + 2] cycloaddition of N-tosylhydrazones with ortho-quinone methides was developed, affording the facile synthesis of diverse 1,3-oxazine derivatives under mild conditions. In this transformation, N-tosylhydrazones are used as a 1,2-dipole synthon under base-free conditions. Moreover, the substrate scope is broad, and the products are formed with high diastereoselectivities in most of the cases.

Boron-Catalyzed Azide Insertion of α-Aryl α-Diazoesters.

A challenging metal-free azide insertion of α-aryl α-diazoesters in the presence of B(C6F5)3 (5 mol %) was developed for the first time. The reaction features an easy operation, wide substrate scope, and mild conditions and affords the corresponding products in moderate to high yields. More importantly, alkene and alkyne functional groups were well tolerated because no cyclopropanation or cyclopropenation was observed. Furthermore, the corresponding azide products could be converted to primary amines or 1,2,3-triazole derivatives after simple transformations.

[Simultaneous determination of 8 constituents of Desmodii Styracifolii Herba in rat bile with HPLC-MS/MS and their excretion pharmacokinetics].

To establish a HPLC-MS/MS quantitative method for analysis of 8 constituents in rat bile. The method was applied in the biliary excretion study after oral administration of Desmodii Styracifolii Herba extract. The rat bile samples were collected during 0-1, 1-2, 2-4, 4-6, 6-8, 8-12, 12-24 h. Diamonsil C18column (4.6 mm×150 mm, 5 µm) was adopted and eluted with methanol and 0.01% acetic acid in a gradient mode. The flow rate was 0.8 mL•min⁻¹, and the column temperature was set at 40 ℃. The detection was carried out by a triple quadrupole linearion trap mass spectrometer in the negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. The calibration curves showed a good linearity, with correlation coefficients (r≥0.991 5) for all of the analytes within the concentration range. The intra-day and inter-day precisions (RSD) were both less than 15%, and the accuracies (RE) ranged from -15% to 15%. The method had a good stability, and was suitable for the content determination of 8 constituents in rat bile. The extraction recoveries of the 8 analytes were more than 63.2%, and the RSD of IS-normalized matrix factors were no more than 15%, which met the requirements for analysis. According to the biliary excretion study, the bile excretion rates of the 8 analytes were relatively low, with great difference among the individuals. The results showed that the established method was simple, selective and specific, thus could be applied in the biliary excretion study of the 8 analytes in rat bile.

Factors controlling the long-term temporal and spatial patterns of nitrate-nitrogen export in a dairy farming watershed.

It is difficult to investigate the factors that control the riverine nitrate-nitrogen (NO3--N) export in a watershed which gains or losses groundwater. To control the NO3--N contamination in these watersheds, it is necessary to investigate the factors that are related to the export of NO3--N that is only produced by the watershed itself. This study was conducted in the Shibetsu watershed located in eastern Hokkaido, Japan, which gains external groundwater contribution (EXT) and 34% of the annual NO3--N loading occurs through EXT. The riverine NO3--N exports from 1980 to 2009 were simulated by the SWAT model, and the factors controlling the temporal and spatial patterns of NO3--N exports were investigated without considering the EXT. The results show that hydrological events control NO3--N export at the seasonal scale, while the hydrological and biogeochemical processes are likely to control NO3--N export at the annual scale. There was an integrated effect among the land use, topography, and soil type related to denitrification process, that regulated the spatial patterns of NO3--N export. The spatial distribution of NO3--N export from hydrologic response units (HRUs) identified the agricultural areas with surplus N that are vulnerable to nitrate contamination. A new standard for the N fertilizer application rate including manure application should be given to control riverine NO3--N export. This study demonstrates that applying the SWAT model is an appropriate method to determine the temporal and spatial patterns of NO3--N export from the watershed which includes EXT and to identify the crucial pollution areas within a watershed in which the management practices can be improved to more effectively control NO3--N export to water bodies.

[A Study of the Relationship Among Genetic Distances, NIR Spectra Distances, and NIR-Based Identification Model Performance of the Seeds of Maize Iinbred Lines].

This paper explored the relationship among genetic distances, NIR spectra distances and NIR-based identification model performance of the seeds of maize inbred lines. Using 3 groups (total 15 pairs) of maize inbred lines whose genetic distaches are different as experimental materials, we calculates the genetic distance between these seeds with SSR markers and uses Euclidean distance between distributed center points of maize NIR spectrum in the PCA space as the distances of NIR spectrum. BPR method is used to build identification model of inbred lines and the identification accuracy is used as a measure of model identification performance. The results showed that, the correlation of genetic distance and spectra distancesis 0.9868, and it has a correlation of 0.9110 with the identification accuracy, which is highly correlated. This means near-Infrared spectrum of seedscan reflect genetic relationship of maize inbred lines. The smaller the genetic distance, the smaller the distance of spectrum, the poorer ability of model to identify. In practical application, near infrared spectrum analysis technology has the potential to be used to analyze maize inbred genetic relations, contributing much to genetic breeding, identification of species, purity sorting and so on. What's more, when creating a NIR-based identification model, the impact of the maize inbred lines which have closer genetic relationship should be fully considered.

[Comparative study on pharmacokinetics and tissue distribution of cis- and trans-2,3,5,4'-Tetrahydroxystilbene-2-O-beta-D-glucosides in mice].

OBJECTIVE: To develop a method for simultaneous determination of cis-2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside (cis-THSG) and trans-2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside (trans-THSG) in mice and comparative study of cis-THSG and trans-THSG in pharmacokinetic and tissue distribution. METHODS: Analyses was performed using a Diamonsil C,18 (250 mm x 4.6 mm, 5 microm) column with acetonitrile-methanol-0.1% acetic acid (11:7: 82) as the mobile phase at a flow rate of 1.0 mL/min. Polydatin was took as the internal standard. The detection wavelength was set at 285 and 320 nm. RESULTS: Intra-day and inter-day RSD were less than 9.47%. The recoveries and stabilities( RE) were ranged from -9.69% to 4.18% and from -9.49% to 1.33%, respectively. The pharmacokinetic parameters showed that the elimination half-life of cis-THSG was longer than that of trans-THSG and the biological availability of cis-THSG was higher than that of trans-THSG. Both cis-THSG and trans-THSG were widely distributed in the tissues. At the same dosage,the tissue concentrations of trans-THSG were greater than those of cis-THSG. CONCLUSION: The method is specific,sensitive, simple, rapid and suitable for simultaneous determination of cis-THSG and trans-THSG in mice. The results of pharmacokinetic and tissue distribution indicated that there was significant difference between cis-THSG and trans-THSG in vivo.

Cell-free DNA testing for early hepatocellular carcinoma surveillance.

BACKGROUND: Liver cirrhosis (LC) is the highest risk factor for hepatocellular carcinoma (HCC) development worldwide. The efficacy of the guideline-recommended surveillance methods for patients with LC remains unpromising. METHODS: A total of 4367 LCs not previously known to have HCC and 510 HCCs from 16 hospitals across 11 provinces of China were recruited in this multi-center, large-scale, cross-sectional study. Participants were divided into Stage Ⅰ cohort (510 HCCs and 2074 LCs) and Stage Ⅱ cohort (2293 LCs) according to their enrollment time and underwent Tri-phasic CT/enhanced MRI, US, AFP, and cell-free DNA (cfDNA). A screening model called PreCar Score was established based on five features of cfDNA using Stage Ⅰ cohort. Surveillance performance of PreCar Score alone or in combination with US/AFP was evaluated in Stage Ⅱ cohort. FINDINGS: PreCar Score showed a significantly higher sensitivity for the detection of early/very early HCC (Barcelona stage A/0) in contrast to US (sensitivity of 51.32% [95% CI: 39.66%-62.84%] at 95.53% [95% CI: 94.62%-96.38%] specificity for PreCar Score; sensitivity of 23.68% [95% CI: 14.99%-35.07%] at 99.37% [95% CI: 98.91%-99.64%] specificity for US) (P < 0.01, Fisher's exact test). PreCar Score plus US further achieved a higher sensitivity of 60.53% at 95.08% specificity for early/very early HCC screening. INTERPRETATION: Our study developed and validated a cfDNA-based screening tool (PreCar Score) for HCC in cohorts at high risk. The combination of PreCar Score and US can serve as a promising and practical strategy for routine HCC care. FUNDING: A full list of funding bodies that contributed to this study can be found in Acknowledgments section.

Influence of RRM, RGG and Potential Phosphorylated Sites in Cold-Inducible Protein RBM3 on its Subcellular Localization and Neuroprotective Effects.

BACKGROUND: As a potent mediator of hypothermic neuroprotection, the cold-inducible protein RBM3 is characterized with one RNA-recognition motifs (RRM) and one arginine-glycine-rich (RGG) domain. It is known that these conserved domains are required for nuclear localization in some RNA-binding proteins. However, little is known about the actual role of RRM and RGG domains in subcellular localization of RBM3. METHODS: To clarify it, various mutants of human Rbm3 gene were constructed. Plasmids were transfected into cells and the localization of RBM3 protein and its varias mutants in cells and role in neuroprotection. RESULTS: In human neuroblastoma SH-SY5Y cells, either a truncation of RRM domain (aa 1-86) or RGG domain (aa 87-157) led to an obvious cytoplasmic distribution, compared to a predominant nuclear localization of whole RBM3 protein (aa 1-157). In contrast, mutants in several potential phosphorylated sites of RBM3, including Ser102, Tyr129, Ser147, and Tyr155, did not alter the nuclear localization of RBM3. Similarly, mutants in two Di-RGG motif sites also did not affect the subcellular distribution of RBM3. Lastly, the role of Di-RGG motif in RGG domains was further investigated. The mutant of double arginines in either Di-RGG motif-1 (Arg87/90) or -2 (Arg99/105) exhibited a higher cytoplasmic localization, indicating that both Di-RGG motifs are required for nucleic localization of RBM3. CONCLUSIONS: Our data suggest that RRM and RGG domains are both required for the nuclear localization of RBM3, with two Di-RGG domain being crucial for nucleocytoplasmic shuttling of RBM3.

The development of a cSMART-based integrated model for hepatocellular carcinoma diagnosis.

BACKGROUND: Hepatocellular carcinoma (HCC) generally arises from a background of liver cirrhosis (LC). Patients with cirrhosis and suspected HCC are recommended to undergo serum biomarker tests and imaging diagnostic evaluation. However, the performance of routine diagnostic methods in detecting early HCC remains unpromising. METHODS: Here, we conducted a large-scale, multicenter study of 1675 participants including 490 healthy controls, 577 LC patients, and 608 HCC patients from nine clinical centers across nine provinces of China, profiled gene mutation signatures of cell-free DNA (cfDNA) using Circulating Single-Molecule Amplification and Resequencing Technology (cSMART) through detecting 931 mutation sites across 21 genes. RESULTS: An integrated diagnostic model called "Combined method" was developed by combining three mutation sites and three serum biomarkers. Combined method outperformed AFP in the diagnosis of HCC, especially early HCC, with sensitivities of 81.25% for all stages and 66.67% for early HCC, respectively. Importantly, the integrated model exhibited high accuracy in differentiating AFP-negative, AFP-L3-negative, and PIVKA-II-negative HCCs from LCs.

Enhancement of photoheterotrophic biohydrogen production at elevated temperatures by the expression of a thermophilic clostridial hydrogenase.

The working temperature of a photobioreactor under sunlight can be elevated above the optimal growth temperature of a microorganism. To improve the biohydrogen productivity of photosynthetic bacteria at higher temperatures, a [FeFe]-hydrogenase gene from the thermophile Clostridium thermocellum was expressed in the mesophile Rhodopseudomonas palustris CGA009 (strain CGA-CThydA) using a log-phase expression promoter P( pckA ) to drive the expression of heterogeneous hydrogenase gene. In contrast, a mesophilic Clostridium acetobutylicum [FeFe]-hydrogenase gene was also constructed and expressed in R. palustris (strain CGA-CAhydA). Both transgenic strains were tested for cell growth, in vivo hydrogen production rate, and in vitro hydrogenase activity at elevated temperatures. Although both CGA-CThydA and CGA-CAhydA strains demonstrated enhanced growth over the vector control at temperatures above 38 °C, CGA-CThydA produced more hydrogen than the other strains. The in vitro hydrogenase activity assay, measured at 40 °C, confirmed that the activity of the CGA-CThydA hydrogenase was higher than the CGA-CAhydA hydrogenase. These results showed that the expression of a thermophilic [FeFe]-hydrogenase in R. palustris increased the growth rate and biohydrogen production at elevated temperatures. This transgenic strategy can be applied to a broad range of purple photosynthetic bacteria used to produce biohydrogen under sunlight.

Association between Sleep and Suicidal Ideation in Chinese Undergraduate Students.

Suicide is an important global public health issue, which deserves more attention. This study aims to examine the relative independent relationship between suicide ideation and subjective sleep quality, sleep hygiene, and insomnia symptoms in undergraduate students in China. This population-based study included 2379 undergraduate students aged 18-26, randomly recruited from three public universities in Shanghai. The participants completed four questionnaires: the Pittsburgh Sleep Quality Index; Sleep Hygiene Practice Scale; Insomnia Severity Index; and the Symptom Checklist 90 (specifically the depression and anxiety dimensions and Q15-suicide ideation). The results of Spearman's correlation analysis indicate that poor sleep quality, short sleep duration, poor sleep hygiene, and insomnia symptoms were all associated with suicidal ideation in undergraduate students. However, according to the results of the hierarchical linear regression, no experience of sharing a bedroom at home, poor relationship with roommates, short sleep duration, sleep medicine use, and good daytime function were related to suicidal ideation, after controlling for the symptoms of depression and anxiety, which may be important in the identification of suicidal ideation. Sleep problems are highly discoverable and modifiable, and have a low sense of shame, therefore, sleep interventions for individuals with suicidal ideation and poor sleep quality may be an efficient and effective approach to suicide prevention.

Long-Term Prognostic Factors in Patients With Antineutrophil Cytoplasmic Antibody-Associated Vasculitis: A 15-Year Multicenter Retrospective Study.

Background: Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a multisystem autoimmune disease with small-vessel involvement. In AAV, microscopic polyangiitis (MPA) and granulomatosis with polyangiitis (GPA) are major clinicopathologic variants. In addition, myeloperoxidase (MPO) and proteinase 3 (PR3) are major target antigens. The objective of the study was to explore the predictive factors for long-term survival in AAV patients. Materials and Methods: A multicenter retrospective study was carried out on 407 patients between 2005 and 2020. Clinical parameters were obtained from laboratory tests including the ANCA types, antinuclear antibody (ANA), extractable nuclear antigen (ENA), anti-streptolysin O (ASO), glomerular filtration rate (GFR), and the laboratory examinations for the blood routine, liver function, renal function, and immunity, etc. The data for clinical parameters were collected from electronic medical records (EMRs), and the data for patient survival were acquired through regular follow-up. The association of clinical parameters with overall survival (OS) along with 3-year and 5-year survival rates was analyzed, and the nomogram as a predictive model was established according to the analysis results. Results: In the present study, 336 (82.6%) patients and 46 (11.3%) patients were diagnosed with MPA and GPA, respectively. The mean and median OS for all the patients were 2,285 and 2,290 days, respectively. The 1-year, 3-year, 5-year, and 10-year cumulative survival rates for all the patients were 84.2%, 76.3%, 57.2%, and 32.4%, respectively. Univariate and multivariate survival analyses indicated that the independent prognostic factors included age, pathological categories (MPA, GPA, and other types), serum ANCA types (negative or positive for MPO and/or PR3), ANA, ASO, GFR, lymphocyte, neutrophil-to-lymphocyte ratio (NLR), and C-reactive protein (CRP), and these clinical parameters except for ASO were used to construct a nomogram. The nomogram for 3-year and 5-year survival rates had a C-index of 0.721 (95% CI 0.676-0.766). The calibration curves showed that the predicted values of the nomogram for 3-year and 5-year survival rates were generally consistent with practical observed values, and decision curve analysis (DCA) further demonstrated the practicability and accuracy of the predictive model. Conclusion: Laboratory tests at diagnosis have great significance in the prediction of long-term survival in AAV patients.

[Dynamic expression of tropomyosin 1 in rat model of hepatic fibrosis and hepatic stellate cells].

OBJECTIVE: To investigate the dynamic expression of TPM1 in rat model of hepatic fibrosis and hepatic stellate cells induced by TGFß1. METHOD: Thirty male SD rats were divided into control group (n = 6) and model group (n = 24). The rat model of hepatic fibrosis was established by intraperitoneal injection of dimethylnitrosamine(DMN). The sera were collected from portal vein and liver tissues were taken from animals 2, 4, 6, 8 weeks HSC-T6 cells were cultured and induced 48 hours by 5 ng/ml TGF-ß1. The pathological changes of liver were observed by Hematoxylin-Eosin and Masson Staining. Reverse Transcription-polymerase chain reaction (RT-PCR), immunohistochemistry and Western-blotting were used to determine the mRNA and protein expressions of TPM1, TGFß1 and α-SMA in rat models and HSC-T6 cells and the localization of TPM1 in rat models. RESULTS: Rat models of hepatic fibrosis were successfully established. TPM1 was lowly stained in the wall of blood vessels in portal areas in normal livers, in fibrotic livers TPM1 was mainly stained along the fibrotic septum. The mRNA and protein expressions of TPM1 and α-SMA in rat models of hepatic fibrosis increased at the week 2 and peaked at week 6, which was statistical significance compared to control group, P < 0.05; TGF-ß1 increased at week 2 and it was higher than the levels in other groups at week 4, which was statistical significance compared to control group P < 0.05; Correlation analysis showed that TPM1 positively correlated with α-SMA and TGF-ß1, rs = 0.688, rs = 0.692, P < 0.01. In HSC-T6, the mRNA expressions of TPM1 and α-SMA increased after being induced by TGF -beta1. compare with control group, the differences were significant, P less than 0.05. CONCLUSION: TPM1 may be playing an important role in the occurrence and development of liver fibrosis. Maybe it could become a potential therapeutic target for hepatic fibrosis.

Metabolome and proteome analyses reveal transcriptional misregulation in glycolysis of engineered E. coli.

Synthetic metabolic pathways are a burden for engineered bacteria, but the underlying mechanisms often remain elusive. Here we show that the misregulated activity of the transcription factor Cra is responsible for the growth burden of glycerol overproducing E. coli. Glycerol production decreases the concentration of fructose-1,6-bisphoshate (FBP), which then activates Cra resulting in the downregulation of glycolytic enzymes and upregulation of gluconeogenesis enzymes. Because cells grow on glucose, the improper activation of gluconeogenesis and the concomitant inhibition of glycolysis likely impairs growth at higher induction of the glycerol pathway. We solve this misregulation by engineering a Cra-binding site in the promoter controlling the expression of the rate limiting enzyme of the glycerol pathway to maintain FBP levels sufficiently high. We show the broad applicability of this approach by engineering Cra-dependent regulation into a set of constitutive and inducible promoters, and use one of them to overproduce carotenoids in E. coli.

Multi-omics Analysis of CRISPRi-Knockdowns Identifies Mechanisms that Buffer Decreases of Enzymes in E. coli Metabolism.

Enzymes maintain metabolism, and their concentration affects cellular fitness: high enzyme levels are costly, and low enzyme levels can limit metabolic flux. Here, we used CRISPR interference (CRISPRi) to study the consequences of decreasing E. coli enzymes below wild-type levels. A pooled CRISPRi screen with 7,177 strains demonstrates that metabolism buffers fitness defects for hours after the induction of CRISPRi. We characterized the metabolome and proteome responses in 30 CRISPRi strains and elucidated three gene-specific buffering mechanisms: ornithine buffered the knockdown of carbamoyl phosphate synthetase (CarAB) by increasing CarAB activity, S-adenosylmethionine buffered the knockdown of homocysteine transmethylase (MetE) by de-repressing expression of the methionine pathway, and 6-phosphogluconate buffered the knockdown of 6-phosphogluconate dehydrogenase (Gnd) by activating a bypass. In total, this work demonstrates that CRISPRi screens can reveal global sources of metabolic robustness and identify local regulatory mechanisms that buffer decreases of specific enzymes. A record of this paper's transparent peer review process is included in the Supplemental Information.

The Landscape of Cell-Free HBV Integrations and Mutations in Cirrhosis and Hepatocellular Carcinoma Patients.

PURPOSE: Intratumoral hepatitis B virus (HBV) integrations and mutations are related to hepatocellular carcinoma (HCC) progression. Circulating cell-free DNA (cfDNA) has shown itself as a powerful noninvasive biomarker for cancer. However, the HBV integration and mutation landscape on cfDNA remains unclear. EXPERIMENTAL DESIGN: A cSMART (Circulating Single-Molecule Amplification and Resequencing Technology)-based method (SIM) was developed to simultaneously investigate HBV integration and mutation landscapes on cfDNA with HBV-specific primers covering the whole HBV genome. Patients with HCC (n = 481) and liver cirrhosis (LC; n = 517) were recruited in the study. RESULTS: A total of 6,861 integration breakpoints including TERT and KMT2B were discovered in HCC cfDNA, more than in LC. The concentration of circulating tumor DNA (ctDNA) was positively correlated with the detection rate of these integration hotspots and total HBV integration events in cfDNA. To track the origin of HBV integrations in cfDNA, whole-genome sequencing (WGS) was performed on their paired tumor tissues. The paired comparison of WGS data from tumor tissues and SIM data from cfDNA confirmed most recurrent integration events in cfDNA originated from tumor tissue. The mutational landscape across the whole HBV genome was first generated for both HBV genotype C and B. A region from nt1100 to nt1500 containing multiple HCC risk mutation sites (OR > 1) was identified as a potential HCC-related mutational hot zone. CONCLUSIONS: Our study provides an in-depth delineation of HBV integration/mutation landscapes at cfDNA level and did a comparative analysis with their paired tissues. These findings shed light on the possibilities of noninvasive detection of virus insertion/mutation.

[Distribution Characteristics and Source Identification of Organophosphate Esters in Key Waters Areas of Taihu Lake].

Organophosphate esters (OPEs) are ubiquitous in the environment and pose potential risks to ecosystems in that they produce cytotoxicity, genetic toxicity, developmental toxicity, neurotoxicity, and the endocrine disrupting effect. This study aimed to investigate the distribution, sources, and ecological risk of thirteen OPEs in industrial wastewater, influents and effluents of wastewater treatment plants, and the surface waters of key areas (Changzhou City) in Taihu Lake. The results showed that ∑OPEs in industrial wastewater ranged from 91.70-840.52 ng·L-1. The profiles varied from different industries; however, tris (1-chloro-2-propyl) phosphate (TCPP), triethyl phosphate (TEP), and tris (2-chloroethyl) phosphate (TCEP) were the dominant compounds. The ∑OPEs from the wastewater treatment plants were relatively higher, ranging from 1859.59-2778.57 ng·L-1. They are rather resistant to traditional wastewater treatment techniques. The removal rate of ∑OPEs, aryl OPEs, and chlorinated OPEs was 14.97%, 97.91%, and 4.37% (for W1) and 17.32%, 90.83%, and 5.40% (for W2), respectively. The concentration of OPEs in the surface water was in the range of 219.47-689.85 ng·L-1, which was lower than that in wastewater. In addition, TEP, TCPP and TCEP predominated in the surface water samples. A principle component analysis (PCA) indicated that the industrial wastewater, emissions of vehicles, and the release of building materials may be the main sources of OPEs in the study area. A risk quotient was used to assess the ecological risk of OPEs to aquatic organisms in the surface water. Most of the OPEs showed low ecological risk, but 2-ethylhexyl diphenyl phosphate (EHDPP) posed a moderate risk in some sites, and the possible ecological effects caused by the OPEs should not be ignored in the key areas of Taihu Lake.

[Pollution Characteristics and Risk Assessment of Typical Organophosphate Esters in Beijing Municipal Wastewater Treatment Plant and the Receiving Water].

Organophosphate esters (OPEs) are ubiquitous in the environment and pose a potential threat to ecosystems and human health. A method for the determination of eight OPEs by ultra-performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS) was established. The recovery rates of eight target compounds with different solid-phase extraction columns, different eluents, and different eluent volumes were compared. The results showed that using ENVI-18 column enrichment, OPEs were eluted with 8 mL acetonitrile containing 25% (volume fraction) dichloromethane, and the labeled recovery rate of the target compound was 92.5%-102.2%. The recoveries of different matrix samples were 88.5%-116.1% and relative standard deviation was 1.7%-9.9%. The concentration range of 8 different detectable organophosphate esters in the effluent of sewage treatment plant is 85.9-235.4 ng·L-1 during the six-day sampling process, permissive river downstream of the six-day ΣOPEs average total concentration was 130.3 ng·L-1, higher than the 119.4 ng·L-1 upstream water concentration, but lower than the sewage treatment plant effluent concentration of total 162.5ng·L-1. The study shows that the sewage treatment plant cannot completely remove OPEs; for triethyl phosphate (TEP) and 3 (2-ethyl hexyl) phosphate ester (TEHP) there exists a negative removal phenomenon, whereas for other OPEs the removal rate was between 14.1% and 84.9%, and the total ΣOPEs removal rate by the sewage plant was 50.0%. The TPhP in the effluent of the sewage treatment plant has medium environmental risk (RQ>0.1), and other organophosphates have low environmental risk (RQ<0.1); however, the long-term mixing effects of organophosphate esters on the ecosystem of the receiving river should not be ignored.