Genome-wide direct quantification of in vivo mutagenesis using high-accuracy paired-end and complementary consensus sequencing.

Error-corrected next-generation sequencing (ecNGS) is an emerging technology for accurately measuring somatic mutations. Here, we report paired-end and complementary consensus sequencing (PECC-Seq), a high-accuracy ecNGS approach for genome-wide somatic mutation detection. We characterize a novel 2-aminoimidazolone lesion besides 7,8-dihydro-8-oxoguanine and the resulting end-repair artifacts originating from NGS library preparation that obscure the sequencing accuracy of NGS. We modify library preparation protocol for the enzymatic removal of end-repair artifacts and improve the accuracy of our previously developed duplex consensus sequencing method. Optimized PECC-Seq shows an error rate of <5 × 10-8 with consensus bases compressed from approximately 25 Gb of raw sequencing data, enabling the accurate detection of low-abundance somatic mutations. We apply PECC-Seq to the quantification of in vivo mutagenesis. Compared with the classic gpt gene mutation assay using gpt delta transgenic mice, PECC-Seq exhibits high sensitivity in quantitatively measuring dose-dependent mutagenesis induced by Aristolochic acid I (AAI). Moreover, PECC-Seq specifically characterizes the distinct genome-wide mutational signatures of AAI, Benzo[a]pyrene, N-Nitroso-N-ethylurea and N-nitrosodiethylamine and reveals the mutational signature of Quinoline in common mouse models. Overall, our findings demonstrate that high-accuracy PECC-Seq is a promising tool for genome-wide somatic mutagenesis quantification and for in vivo mutagenicity testing.

Open versus robot-assisted retroperitoneal tumors resection involving inferior vena cava, abdominal aorta, and renal hilum: a comparative study.

INTRODUCTION: Surgery is currently the only effective treatment for retroperitoneal tumors that do not involve any specific organ. The use of robots for removing both benign and malignant retroperitoneal tumors is considered safe and feasible. However, there is insufficient evidence to determine whether robotic retroperitoneal tumor resection (RMBRs) is superior to open retroperitoneal malignant resection (OMBRs). This study compares the short-term outcomes of robotic excision of benign and malignant retroperitoneal tumors with open excision of the same-sized tumors. METHODS: The study compared demographics and outcomes of patients who underwent robotic resection (n = 54) vs open resection (n = 54) of retroperitoneal tumors between March 2018 and December 2022. A 1:1 matching analysis was conducted to ensure a fair comparison. RESULTS: The study found that RBMRs resulted in reduced operative time (OT), estimated blood loss (EBM), and postoperative hospital stay (PSH) when compared to OBMRs. Additionally, RBMRs reduced EBL, PHS, and OT for patients with malignant tumor involvement in major vessels. No significant differences were found in tumor size, blood transfusion rate, and morbidity rate between the RBMRs and OBMRs groups. CONCLUSION: When comparing RMBRs to OMBRs, it was observed that RMBR was associated with lower (EBL), shorter postoperative hospital stays (PHS), and reduced operative time (OT) in a specific group of patients with both benign and malignant tumors.

Application of next-generation sequencing in the detection of low-abundance mutations.

Mutation accumulation in somatic cells contributes to cancer development, aging and many non-malignant diseases. The true mutation frequency in normal cells is extremely low, which presents a challenge in detecting these mutations at such low frequencies. The emergence of next-generation sequencing (NGS) technology enables direct detection of rare mutations across the entire genome of any species. This breakthrough overcomes numerous limitations of traditional mutation detection techniques that rely on specific detection models and sites. However, conventional NGS is limited in its application for detecting low-frequency mutations due to its high sequencing error rate. To address this challenge, high-accuracy NGS sequencing techniques based on molecular consensus sequencing strategies have been developed. These techniques have the ability to correct sequencing errors, resulting in error rates lower than 10-7, are expected to serve as effective tools for low-frequency mutation detection. Error-corrected NGS (ecNGS) techniques hold great potential in various areas, including safety evaluation and research on environmental mutagens, risk assessment of cell and gene therapy drugs, population health risk monitoring, and fundamental research in life sciences. This review highlights a comprehensive review of the research progress in low-frequency mutation detection techniques based on NGS, and provides a glimpse into their potential applications. It also offers an outlook on the potential applications of these techniques, thereby providing valuable insights for further development, research, and application of this technology in relevant fields.

The whole-cell proteome shows the characteristics of macrolides-resistant Bordetella pertussis in China linked to the biofilm formation.

The macrolides-resistant Bordetella pertussis (MR-Bp) isolates in China evolved from the ptxP1/fhaB3 allele and rapidly became predominant, suggestive of an adaptive transmission ability. This was different from the global prevalent ptxP3 strains, in which MR-Bp was rarely reported. The study aimed to determine the underlying mechanism responsible for fitness and resistance in these two strains. We identify proteomic differences between ptxP1/fhaB3 and ptxP3/fhaB1 strains using tandem mass tag (TMT)-based proteomics. We then performed in-depth bioinformatic analysis to determine differentially expressed genes (DEGs), followed by gene ontology (GO), and protein-protein interaction (PPI) network analysis. Further parallel reaction monitoring (PRM) analysis confirmed the expression of four target proteins. Finally, the crystal violet method was used to determine biofilm-forming ability. The results showed that the main significantly different proteins between the two represent isolates were related to biofilm formation. Furthermore, we have confirmed that ptxP1/fhaB3 showed hyperbiofilm formation in comparison with ptxP3/fhaB1. It is suggested that the resistance and adaptability of ptxP1/fhaB3 strains may be related to the formation of biofilm through proteomics. In a word, we determined the significantly different proteins between the ptxP1/fhaB3 and ptxP3/fhaB1 strains through whole-cell proteome, which were related to biofilm formation.

[Mendelian randomization of diabetes and prostate cancer risk in East Asian population].

OBJECTIVE: Mendelian randomization (MR) was used to explore the causal relationship between diabetes (type 1 and type 2) and prostate cancer (PCa) in East Asian population. METHODS: Mendelian randomization is a causal inference method based on genetic variation, which uses the influence of randomly assigned genotypes in nature on phenotype to infer the impact of biological factors on diseases. This study used genetic variation genes related to inflammatory biomarkers as instrumental variables to improve inference, and patient data was obtained from the GWAS database's aggregated association results. In the individual sample, we estimated the correlation between instrumental variables (SNPs) and type 1 and type 2 diabetes, and screened out strongly related SNPs, and excluded SNPs related to prostate cancer. After screening, further sensitivity analysis and visualization of research results were carried out to test the blood glucose level and the causal relationship between diabetes and prostate cancer. RESULTS: Our MR analysis found that there was a negative causal relationship between the risk of prostate cancer and type 2 diabetes.The ratio of type 2 diabetes to prostate cancer causal relationship (OR)=1.0039, 95% confidence interval (CI)=(1.0008, 1.0071), P=0.013, while type 1 diabetes had fewer SNPs screened, Failed to conduct relevant follow-up analysis. CONCLUSIONS: Under Mendel's randomization hypothesis, our research results show that in the East East Asian population, the gene predicted type 2 diabetes and the occurrence of prostate cancer have a negative causal relationship, while the causal relationship between type 1 diabetes and prostate cancer is unknown due to the limited number of instrumental variables.

Application of 16 G Biopsy Needle in Transperineal Template-Guided Prostate Biopsy.

BACKGROUND: The aim of this study was to evaluate the clinical value of 16 G biopsy needle in transperineal template-guided prostate biopsy (TTPB), compared with 18 G biopsy needle. METHODS: The patients who underwent TTPB from August 2020 to February 2021 were randomized into 2 groups using a random number table. The control group (n = 65) and the observation group (n = 58) performed biopsy with 18 G (Bard MC l820) and 16 G (Bard MC l616) biopsy needles, respectively. Positive rate of biopsy, Gleason score, complications, and pain score were statistically analyzed. RESULTS: The age, prostate volume, PSA, and the number of cores were comparable between the 2 groups. The positive rate of biopsy in the observation group was 68.9% (40/58), meanwhile the control group was 46.2% (30/65). There was statistical difference between the 2 groups (p = 0.011). Gleason score of the observation group (8 [7-9]) was higher than that of the control group (8 [6-9]) (p = 0.038). There was no significant difference in pain score and complications including hematuria, hematospermia, perineal hematoma, infection, and urinary retention between the 2 groups (p > 0.05). CONCLUSIONS: 16 G biopsy needle significantly improved the positive rates and accurately evaluate the nature of lesions, meanwhile did not increase the incidence of complications compared with 18 G biopsy needle.

A high-throughput microplate toxicity screening platform based on Caenorhabditis elegans.

Caenorhabditis elegans (C. elegans), an established model organism, has been widely used in environmental toxicology research. However, most of the current toxicity testing methods based on worms are time-consuming. In this study we aimed to develop an automated and highly-integrated platform for high-throughput and in situ toxicity testing. Considering the superiority of C. elegans as a neurotoxicological model, this platform mainly evaluates general toxicology and neurotoxicology endpoints, which are usually induced by metals and pesticides, the major environmental contaminants. Microplates were used as a worm culturing system, which have good compatibility with any commercial microplate applicable instruments. We developed a microfluidic-based module for worm dispensing, and an image acquisition/analysis module for monitoring worms and detecting toxicity endpoints in bright filed. These were collectively incorporated with a commercial pipetting workstation for automated food/drug delivery and a high-content analysis system for fluorescence detection. The integrated platform achieved an efficient on-demand worm dispensing, long-term maintenance, regular monitoring and imaging, survival assay and behavioral analyses, and visualized gene reporter assay. Moreover, "Lab on Web" was achieved by connecting the platform to the web for remote operation, worm monitoring, and phenotype calculation. To demonstrate the ability of the platform for automated toxicity testing assays; worms were treated with cadmium and longevity, neurotoxicity, developmental toxicity and gst-4 expression were evaluated. We determined its feasibility and proposed the potential application in high-throughput toxicity screening for environmental risk assessment in the nearest future.

Comparison of the Oncological and Functional Outcomes of Brachytherapy and Radical Prostatectomy for Localized Prostate Cancer.

Background and Objectives: To compare the oncological and functional outcomes of brachytherapy (BT) and radical prostatectomy (RP) in patients with localized prostate cancer (PCa). Materials and Methods: We retrospectively analyzed data from 557 patients with localized PCa who were treated with BT (n = 245) or RP (n = 312) at Northern Jiangsu People's Hospital between January 2012 and December 2017. Biochemical relapse-free survival (bRFS) and cancer-specific survival (CSS) were compared by treatment modality. Multivariate Cox regression analysis was used to evaluate bRFS. Health-related quality of life (HRQoL) was measured using the Expanded Prostate Cancer Index Composite (EPIC) questionnaire. Results: The BT group was older and had a higher initial PSA (iPSA). The 5-year bRFS was 82.9% in the BT group versus 80.1% in the RP group (p = 0.570). The 5-year CSS was 96.4% in the BT group versus 96.8% in the RP group (p = 0.967). Based on multivariate Cox regression analysis, Gleason score ≥ 8 was the main independent prognostic factor for bRFS. Regarding the HRQoL, compared with the baseline, both treatments produced a significant decrease in different aspects of HRQoL at 3, 6, and 12 months after treatment. Patients in the BT group had lower HRQoL with regard to urinary irritation/obstruction and bowel function or bother, while patients in the RP group had lower HRQoL concerning urinary incontinence and sexual function or bother. There was no significant difference in HRQoL aspects between the two groups after follow-up for 2 years compared with the baseline. Conclusions: BT provides equivalent oncological control outcomes in terms of bRFS and CSS for patients with localized PCa compared with RP. Gleason score ≥ 8 was the main independent prognostic factor for bRFS. BT had better HRQoL compared with RP, except for urinary irritation/obstruction and bowel function or bother, but returned to baseline after 2 years.

The Efficacy of Urinary Continence in Patients Undergoing Robot-Assisted Radical Prostatectomy with Bladder-Prostatic Muscle Reconstruction and Bladder Neck Eversion Anastomosis.

Background and Objectives: To evaluate the efficacy of bladder-prostatic muscle reconstruction and bladder neck eversion anastomosis in the recovery of urinary continence after robot-assisted radical prostatectomy (RARP). Materials and Methods: From January 2020 to May 2022, 69 patients who underwent RARP in our hospital were recruited. Thirty-seven patients underwent RARP with the Veil of Aphrodite technique (control group). On the basis of the control group, 32 patients underwent bladder-prostatic muscle reconstruction and bladder neck eversion anastomosis during RARP (observation group). The recovery of urinary continence was followed up at 24 h and 1, 4, 12, and 24 weeks after catheter removal. Results: There were no significant differences in operative time (127.76 ± 21.23 min vs. 118.85 ± 24.71 min), blood loss (118.27 ± 16.75 mL vs. 110.77 ± 19.63 mL), rate of leakage (3.13% vs. 2.70%), rate of positive surgical margin (6.25% vs. 10.81%), or postoperative Gleason score [7 (6−8) vs. 7 (7−8)] between the observation group and the control group (p > 0.05). After catheter removal, the rates of urinary continence at 24 h, 1 week, 4 weeks, 12 weeks, and 24 weeks were 46.88%, 68.75%, 84.38%, 90.63%, and 93.75% in the observation group, respectively. Meanwhile, the rates of urinary continence in the control group were 21.62%, 37.84%, 62.16%, 86.49%, and 91.89%, respectively. There was a significant difference between the two groups (p = 0.034), especially at 24 h, 1 week, and 4 weeks after catheter removal (p < 0.05). Conclusions: Bladder-prostatic muscle reconstruction and bladder neck eversion anastomosis were beneficial to the recovery of urinary continence after RARP, especially early urinary continence.

Photocatalytic Fuel Cell-Assisted Molecularly Imprinted Self-Powered Sensor: A Flexible and Sensitive Tool for Detecting Aflatoxin B1.

The self-powered electrochemical sensor has gained big achievements in energy and devices, but it is challenging in analytical application owing to its low energy conversion efficiency and limited selectivity caused by the plentiful interference in actual samples. Herein, a new self-powered biosensor was constructed by the integration of a photocatalytic fuel cell (PFC) with a molecular imprinting polymer (MIP) to achieve sensitive and specific detection of aflatoxin B1 (AFB1). Compared with other fuel cells, the PFC owns the advantages of low cost, high energy, good stability, and friendly environment by using light as the excitation source. MoS2-Ti3C2Tx MXene (MoS2-MX) served as the photoanode material for the first time by forming a heterojunction structure, which can enhance the photocurrent by about 3-fold and greatly improve the photoelectric conversion efficiency. Aiming at the poor selectivity of the self-powered sensor, the MIP was introduced to achieve the specific capture and separation of targets without sample pretreatment. Using the MIP and PFC as recognition and signal conversion elements, respectively, the proposed self-powered biosensor showed a wide dynamic range of 0.01-1000 ng/mL with a detection limit of 0.73 pg/mL, which opened opportunities to design more novel self-powered biosensors and promoted its application in food safety and environmental monitoring.

Benchmark dose analysis of multiple genotoxicity endpoints in gpt delta mice exposed to aristolochic acid I.

As the carcinogenic risk of herbs containing aristolochic acids (AAs) is a global health issue, quantitative evaluation of toxicity is needed for the regulatory decision-making and risk assessment of AAs. In this study, we selected AA I (AAI), the most abundant and representative compound in AAs, to treat transgenic gpt delta mice at six gradient doses ranging from 0.125 to 4 mg/kg/day for 28 days. AAI-DNA adduct frequencies and gpt gene mutation frequencies (MFs) in the kidney, as well as Pig-a gene MFs and micronucleated reticulocytes (MN-RETs) frequencies in peripheral blood, were monitored. The dose-response (DR) relationship data for these in vivo genotoxicity endpoints were quantitatively evaluated using an advanced benchmark dose (BMD) approach with different critical effect sizes (CESs; i.e., BMD5, BMD10, BMD50 and BMD100). The results showed that the AAI-DNA adduct frequencies, gpt MFs and the MN-RETs presented good DR relationship to the administrated doses, and the corresponding BMDL100 (the lower 90% confidence interval of the BMD100) values were 0.017, 0.509 and 3.9 mg/kg/day, respectively. No positive responses were observed in the Pig-a MFs due to bone marrow suppression caused by AAI. Overall, we quantitatively evaluated the genotoxicity of AAI at low doses for multiple endpoints for the first time. Comparisons of BMD100 values across different endpoints provide a basis for the risk assessment and regulatory decision-making of AAs and are also valuable for understanding the genotoxicity mechanism of AAs.

Risk factors for infection complications after transrectal ultrasound-guided transperineal prostate biopsy.

BACKGROUND: To investigate the influence factors of infection complications of transrectal ultrasound-guided transperineal prostate biopsy. METHODS: A total of 2192 patients who underwent prostate biopsy under transperineal prostate biopsy were analyzed retrospectively from December 2010 to May 2020.We collected the clinical characteristics and the incidence of complications, and used univariate and multivariate logistic regression analyses to analyze independent risk factors for infection complications after transperineal prostate biopsy. RESULTS: Univariate analysis showed that the following factors were associated with the infection complications: diabetes, bacterial prostatitis, history of urinary retention, history of urinary infection, and number of cores. Furthermore, multivariate logistic analysis revealed that diabetes (OR 2.037, 95% CI 1.143-3.572, P = 0.021) and history of urinary retention (OR 2.563, 95% CI 1.284-3.901, P = 0.013) were independent risk factors for infection complications after transperineal prostate biopsy. CONCLUSIONS: Patients with diabetes and history of urinary retention were more likely to have infection complications after transperineal prostate biopsy.

Aristolochic acid IVa forms DNA adducts in vitro but is non-genotoxic in vivo.

Aristolochic acids (AAs) are a family of natural compounds with AA I and AA II being known carcinogens, whose bioactivation causes DNA adducts formation. However, other congeners have rarely been investigated. This study aimed to investigate genotoxicity of AA IVa, which differs from AA I by a hydroxyl group, abundant in Aristolochiaceae plants. AA IVa reacted with 2'-deoxyadenosine (dA) and 2'-deoxyguanosine (dG) to form three dA and five dG adducts as identified by high-resolution mass spectrometry, among which two dA and three dG adducts were detected in reactions of AA IVa with calf thymus DNA (CT DNA). However, no DNA adducts were detected in the kidney, liver, and forestomach of orally dosed mice at 40 mg/kg/day for 2 days, and bone marrow micronucleus assay also yielded negative results. Pharmacokinetic analyses of metabolites in plasma indicated that AA IVa was mainly O-demethylated to produce a metabolite with two hydroxyl groups, probably facilitating its excretion. Meanwhile, no reduced metabolites were detected. The competitive reaction of AA I and AA IVa with CT DNA, with adducts levels varying with pH of reaction revealed that AA IVa was significantly less reactive than AA I, probably by hydroxyl deprotonation of AA IVa, which was explained by theoretical calculations for reaction barriers, energy levels of the molecular orbits, and charges at the reaction sites. In brief, although it could form DNA adducts in vitro, AA IVa was non-genotoxic in vivo, which was attributed to its low reactivity and biotransformation into an easily excreted metabolite rather than bioactivation.

Influence of Different Inactivation Methods on Severe Acute Respiratory Syndrome Coronavirus 2 RNA Copy Number.

The outbreak of coronavirus disease 2019 (COVID-19) has spread across the world and was characterized as a pandemic. To protect medical laboratory personnel from infection, most laboratories inactivate the virus causing COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in clinical samples before testing. However, the effect of inactivation on the detection results remains unknown. Here, we used a digital PCR assay to determine the absolute SARS-CoV-2 RNA copy number in 63 nasopharyngeal swab samples and assess the effect of inactivation methods on viral RNA copy number. Viral inactivation was performed by three different methods: (i) incubation with the TRIzol LS reagent for 10 min at room temperature, (ii) heating in a water bath at 56°C for 30 min, and (iii) high-temperature treatment, including autoclaving at 121°C for 20 min, boiling at 100°C for 20 min, and heating at 80°C for 20 min. Compared to the amount of RNA in the original sample, TRIzol treatment destroyed 47.54% of the nucleocapsid protein (N) gene and 39.85% of open reading frame (ORF) 1ab. For samples treated at 56°C for 30 min, the copy number of the N gene and ORF 1ab was reduced by 48.55% and 56.40%, respectively. The viral RNA copy number dropped by 50 to 66% after heating at 80°C for 20 min. Nearly no viral RNA was detected after autoclaving at 121°C or boiling at 100°C for 20 min. These results indicate that inactivation reduced the quantity of detectable viral RNA and may cause false-negative results, especially in weakly positive cases. Thus, use of the TRIzol reagent rather than heat inactivation is recommended for sample inactivation, as the TRIzol reagent had the least effect on the RNA copy number among the tested methods.

The global prevalence ptxP3 lineage of Bordetella pertussis was rare in young children with the co-purified aPV vaccination: a 5 years retrospective study.

BACKGROUND: The global prevalent ptxP3 strains varies from about 10% to about 50% of circulating B. pertussis population in different areas of China. METHODS: To investigate the difference of vaccination status between different genotypes in the circulating B. pertussis after 10 years of acellular pertussis vaccine (aPV) used in China. The nasopharyngeal swabs and isolates of B. pertussis from these patients were used to perform genotyping of antigen genes. We use antibiotic susceptibility test against erythromycin and sequencing methods for site 2047 of 23S rRNA to determine the resistance status. RESULTS: The ptxP1 allele with erythromycin resistant (ER) B. pertussis infection (total of 449 subjects) consisted of 84.70 to 96.70% from 2012 to 2016 in this study. Vaccinated with co-purified aPV was found in 133(133/403,33.0%), 1(1/9,11.1%) and 2(2/21,9.5%) in ptxP1/fhaB3-ER, ptxP1/fhaB2-ES and ptxP3/fhaB2-ES B. pertussis infected children each, which showed a significant difference (χ2 = 6.87, P = 0.032). CONCLUSIONS: The ptxP3-ES B. pertussis was rare while the ptxP1-ER B. pertussis was steadily increased in Xi'an, China from 2012 to 2016, where co-purified aPV was prevalent used. This pose a hypothesis that the co-purified aPV might protect against ptxP3 strains more efficient, which generated a rare chance for ptxP3 strains to be under the antibiotic pressure and further developed to be erythromycin resistance. A further cohort study and the mechanisms of the additional antigen proteins of co-purified aPV protected against B. pertussis should be consideration.

Detection of genome-wide low-frequency mutations with Paired-End and Complementary Consensus Sequencing (PECC-Seq) revealed end-repair-derived artifacts as residual errors.

To improve the accuracy and the cost-efficiency of next-generation sequencing in ultralow-frequency mutation detection, we developed the Paired-End and Complementary Consensus Sequencing (PECC-Seq), a PCR-free duplex consensus sequencing approach. PECC-Seq employed shear points as endogenous barcodes to identify consensus sequences from the overlap in the shortened, complementary DNA strand-derived paired-end reads for sequencing error correction. With the high accuracy of PECC-Seq, we identified the characteristic base substitution errors introduced by the end-repair process of mechanical fragmentation-based library preparations, which were prominent at the terminal 7 bp of the library fragments in the 5'-NpCpA-3' and 5'-NpCpT-3' trinucleotide context. As demonstrated at the human genome scale (TK6 cells), after removing these potential end-repair artifacts from the terminal 7 bp, PECC-Seq could reduce the sequencing error frequency to mid-10-7 with a relatively low sequencing depth. For TA base pairs, the background error rate could be suppressed to mid-10-8. In mutagen-treated (6 µg/mL methyl methanesulfonate or 12 µg/mL N-nitroso-N-ethylurea) TK6, increases in mutagen treatment-related mutant frequencies could be detected, indicating the potential of PECC-Seq in detecting genome-wide ultra-rare mutations. In addition, our finding on the patterns of end-repair artifacts may provide new insights into further reducing technical errors not only for PECC-Seq, but also for other next-generation sequencing techniques.

Genotoxic effects of imidacloprid in human lymphoblastoid TK6 cells.

Among neonicotinoid insecticides, the fastest growing class of insecticides worldwide over the past decade, imidacloprid (IMI) is the most widely used one. The effects of IMI on human health, especially on genetic toxicity have gradually aroused more attention. In this study, a combined in vitro approach employing the thymidine kinase (TK) gene mutation assay, the comet assay and the micronucleus test was taken to assess the genotoxicity of IMI. The mechanism behind IMI was also explored by measuring reactive oxygen species (ROS) in the human lymphoblastoid TK6 cells. The cells were treated with 0.01, 0.1, 1, 5, and 10 µg/mL IMI, and ROS generation was measured by the use of 2,7,-dichlorofluorescin diacetate (DCFH-DA) assay. IMI significantly increased the micronucleus (MN) frequency, TK mutations and DNA damage with a dose-effect relationship, and the lowest effective concentration in those tests was 0.1 µg/mL. However, no obvious change of intracellular ROS was observed for any concentrations. The results indicate that IMI has potential genotoxic effects on TK6 cells, but ROS does not seem to be involved as a mechanism of genotoxicity under the experimental conditions.

An Adaption of Human-Induced Hepatocytes to In Vitro Genetic Toxicity Tests.

Metabolic activation is essential in standard in vitro genotoxicity test systems. At present, there is a lack of suitable cell models that can express the major characteristics of liver function for predicting substance toxicity in humans. Human-induced hepatocytes (hiHeps), which have been generated from fibroblasts by lentiviral expression of liver transcription factors, can express hepatic gene programs and can be expanded in vitro and display functional characteristics of mature hepatocytes, including cytochrome P450 enzyme activity and biliary drug clearance. Our purpose was to investigate whether hiHeps could be used as a more suitable model for genotoxicity evaluation of chemicals. Therefore, a direct mutagen, methylmethanesulfonate (MMS), and five promutagens [2-nitrofluorene (2-NF), benzo[a]pyrene (B[a]P), aflatoxin B1, cyclophosphamide and N-nitrosodiethylamine] were tested by the cytokinesis-block micronucleus test and the comet assay. Results from genotoxicity tests showed that the micronucleus frequencies were significantly increased by all of the six clastogens tested. Moreover, MMS, 2-NF and B[a]P induced significant increases in the % Tail DNA in the comet assay. In conclusion, our findings from the preliminary study demonstrated that hiHeps could detect the genotoxicity of indirect carcinogens, suggesting their potential to be applied as an effective tool for in vitro genotoxicity assessments.

Pelvic plexus block to provide better anesthesia in transperineal template-guided prostate biopsy: a randomised controlled trial.

BACKGROUND: To evaluate the efficacy of pelvic plexus block (PPB) in relief pain during transperineal template-guided prostate biopsy (TTPB), compared with conventional periprostatic nerve block (PNB). METHODS: From July 2016 to August 2017, 245 patients who were performed TTPB in Clinical Medical College of Yangzhou University were recruited. The patients were randomized into three groups using a random number table. Group-1 received prostate capsule local anesthesia with 22 ml of 1% lidocaine. Group-2 additionally received PNB on the basis of Group-1. To perform PNB, 5 ml 1% lidocaine was injected into the region of prostatic neurovascular bundle situated in the angle of prostate-bladder-seminal vesicle. Group-3 received prostate capsule local anesthesia plus PPB (5 ml of 1% lidocaine injection into the pelvic plexus which located on lateral to the bilateral seminal vesicle apex). The patients' pain and satisfaction were evaluated by visual analogue scale and visual numerical scale, respectively. RESULTS: The age, total prostate volume, PSA and the number of cores were comparable among the three groups. The visual analog scale scores of group-3 were significantly lower than group-2 during biopsy (P = 0.003). Conversely, the visual numeric scale scores were higher in group-3 (P = 0.039). Both the group-2 and group-3 outperformed the group-1 in alleviating pain and had a higher quantification of satisfaction. There were no significant differences in the pain scores or the satisfaction scores at 30 min after the procedure among the three groups. CONCLUSIONS: The analgesic efficacy of PPB guided by Doppler ultrasound in TTPB was better than that of PNB and both were superior to no nerve block. TRIAL REGISTRATION: ChiCTR-IOR-17013533 , 01/06/2016.

Secondary Management for Recurrent Ureteropelvic Junction Obstruction after Pyeloplasty: A Comparison of Re-Do Robot-Assisted Laparoscopic Pyeloplasty and Conventional Laparoscopic Pyeloplasty.

OBJECTIVE: To describe and analyze our experience with secondary robot-assisted laparoscopic pyeloplasty (RALP) and conventional laparoscopic pyeloplasty (LP) in treating recurrent ureteropelvic junction obstruction (UPJO) after primary pyeloplasty. METHODS: Patients who underwent secondary RALP or LP for recurrent UPJO were retrospectively analyzed. Baseline characteristics, detailed history of previous pyeloplasty, operative profile and follow-up data were collected and analyzed. RESULTS: Among 29 patients presented with recurrent UPJO, 15 patients underwent secondary RALP. Both groups (RALP or LP) were comparable in baseline characteristics and detailed history of previous pyeloplasty. The mean operative time, suturing time, and hospitalization time of patients in RALP group were significantly less than those in LP group. The mean operative time of RALP group was 2.1 h, while the mean operative time of LP group was 3.23 h. The average suturing time of LP (62.43 min) is about 3 times that of RALP (21.47 min). The overall mean follow-up data was 23 months. The success rate of the RALP group and LP group was 87.7 and 85.7% respectively. CONCLUSION: Compared to LP, RALP may be a better choice for the treatment of recurrent UPJO. Further high-quality clinical studies are needed to confirm the superior nature of RALP.