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UDP-glucuronosyltransferases (UGTs) are the main phase II drug-metabolizing enzymes mediating the most extensive glucuronidation-binding reaction in the human body. The UGT1A family is involved in more than half of glucuronidation reactions. However, significant differences exist in the distribution of UGT1As in vivo and the expression of UGT1As among individuals, and these differences are related to the occurrence of disease and differences in metabolism. In addition to genetic polymorphisms, there is now interest in the contribution of epigenetics and noncoding RNAs (especially miRNAs) to this differential change. Epigenetics regulates UGT1As pretranscriptionally through DNA methylation and histone modification, and miRNAs are considered the key mechanism of posttranscriptional regulation of UGT1As. Both epigenetic inheritance and miRNAs are involved in the differences in sex expression and in vivo distribution of UGT1As. Moreover, epigenetic changes early in life have been shown to affect gene expression throughout life. Here, we review and summarize the current regulatory role of epigenetics in the UGT1A family and discuss the relationship among epigenetics and UGT1A-related diseases and treatment, with references for future research.

Background: Pulsed radiofrequency (PRF) is a minimally invasive interventional technique that provides a novel and effective treatment strategy for neuropathic pain (NP). PRF is advantageous because it does not damage nerves and avoids sensory loss after treatment. At present, animal studies have demonstrated that PRF is safe and effective for relieving the NP associated with sciatic nerve damage in rats with chronic constriction injury (CCI). However, the mechanism through which this effect occurs is unknown. An increasing body of evidence shows that the expression of the P2X ligand-gated ion channel 3 (P2X3) receptor is closely related to NP; this study was to investigate whether the expression of this receptor is involved in NP relief due to PRF. Methods: A total of 36 healthy adult male Sprague-Dawley (SD) rats were randomly divided into three groups: Sham group, CCI group, and PRF group. The right sciatic nerve was ligated in CCI group and PRF group to establish a CCI model; the right sciatic nerve was separated but not ligated in Sham group. On day 14 after the operation, PRF was administered to the ligated sciatic nerve in PRF group (42°C, 45 V, 2 min). A non-live electrode was placed at the exposed sciatic nerve for the rats in Sham and CCI groups. The hindpaw withdrawal threshold (HWT) and thermal withdrawal latency (TWL) were measured at the right hindpaw at different time points before and after PRF or sham therapy. On day 28 after treatment, the dorsal root ganglion (DRG) and spinal dorsal horn of the right L4–6 were harvested from each group to determine the mRNA and protein levels of the P2X3 receptor. Results: On day 28 after PRF treatment, the HWT (8.33 ± 0.67 g vs. 3.62 ± 0.48 g) and TWL (25.42 ± 1.90 s vs. 15.10 ± 1.71 s) were significantly higher in PRF group as compared to CCI group (P < 0.05). The mRNA expression of the P2X3 receptor in the DRG in PRF group was 23.7% lower than that in CCI group (P < 0.05), in the spinal dorsal horns in PRF group was 22.7% lower than that in CCI group (P < 0.05). The protein expression of the P2X3 receptor in the DRG in PRF group was 27.8% lower than that in CCI group (P < 0.05), in the spinal dorsal horns in PRF group was 35.6% lower than that in CCI group (P < 0.05). Conclusion: PRF possibly reduces NP in CCI rats by inhibiting the expression of the P2X3 receptor in the L4–6 DRG and spinal dorsal horns. Key words: Pulsed radiofrequency; P2X3 receptor; Neuropathic pain; Chronic constriction injury

IntroductionTrigeminal neuralgia (TN) is characterised by a sudden, severe, electric shock like paroxysmal pain, which is almost always associated with triggers. Carbamazepine is the first-line medical management of TN. However, side effects are common. Currently, there is no ideal treatment for TN. Since there is a known abnormality of Na+ channels in the trigger zone, 5% lidocaine-medicated plaster (LMP), which can block the Na+ channels on Aδ and C fibres, is an effective treatment method in many chronic pain conditions. A case report has found the benefit of LMP for the treatment of TN without any side effects. Whether LMP is an option for the treatment of TN is worth exploring.Methods and analysisThe PATCH trial is a double-blind, enriched enrolment with randomised withdrawal, vehicle-controlled trial, aiming to explore the effects and safety of LMP in patients with TN. There is a 3-week initial open-label phase, followed by a 4-week double-blind treatment phase for responders. In the double-blind phase, patients will have to withdraw from this PATCH study if they meet one of the following criteria for treatment failure such as: >50% increase in pain intensity or paroxysms, lack of efficacy or side effects. The primary outcome will be the number of treatment failures. Adverse events will also be monitored throughout the study.Ethics and disseminationThis study protocol has been approved by the Institutional Review Board of Beijing Tiantan Hospital (approval number: KY 2020-102-02). The results will be disseminated in international academic meetings and published in peer-reviewed journals.Trial registration numberNCT04570293.

PURPOSE We aimed to analyze the psychological status in breast cancer (BC) patients in the epicenter of the COVID-19 pandemic. PATIENTS AND METHODS A total of 658 individuals were recruited from multiple breast cancer centers in Hubei Province. Online questionnaires were conducted, and included demographic information, clinical features, and four Patient Reported Outcomes (PROs) scales (GAP-7, PHQ-9, ISI, and IES-R). Multivariable logistic regression analysis was designed to identify potential factors on mental health outcomes. RESULTS Questionnaires were collected from February 16 to 19, 2020, the peak time point of COVID-19 outbreak in China. 46.2% BC patients had to modify planned necessary anti-cancer treatment during the outbreak. 8.9% and 9.3% patients reported severe anxiety and severe depression, respectively. 20.8% and 4.0% patients showed severe distress and insomnia, respectively. Multivariable logistic regression analysis demonstrated poor general condition, shorter duration after BC diagnosis, aggressive BC molecular subtypes and close contact with COVID-19 patients as independent factors associated with anxiety. Poor general condition and CVC (central venous catheter) flushing delay were independently association factors depression. In terms of insomnia, poor generation condition was the only associated independent factor. Poor physical condition and treatment discontinuation were underlying risk factors for distress based on multivariable analysis. CONCLUSION High rates of anxiety, depression, distress, and insomnia were observed in patients with breast cancer during the COVID-19 outbreak. Special attention should be paid to psychological status of breast cancer patients especially those with poor general condition, treatment discontinuation, aggressive molecular subtypes and metastatic breast cancer., During the COVID-19 outbreak in China, fear about COVID-19, together with worry about progression of cancer, cause strong emotion stress in cancer patients. We evaluated patient reported outcome in 658 breast cancer patients and survivors recruited from multiple breast cancer centers in Hubei Province using four standardized assessment scales. Multivariable logistic regression analysis was used to identify potential affecting factors on mental health outcomes. High rates of anxiety, depression, distress, and insomnia were observed in patients with breast cancer during the COVID-19 outbreak. Based on our results, living in Wuhan, poor general condition by self-identification, shorter duration after breast cancer diagnosis, aggressive breast cancer molecular subtypes, metastatic breast cancer clinical stage, treatment discontinuation, CVC flushing delay, or close contact with COVID-19 patients are associated risk factors for poorer psychological status. Special attention should be paid to psychological status of breast cancer patients especially those with poor general condition, treatment discontinuation, aggressive molecular subtypes and metastatic breast cancer.

Premature ovarian insufficiency (POI) is a severe clinical syndrome defined by ovarian dysfunction in women less than 40 years old who generally manifest with infertility, menstrual disturbance, elevated gonadotrophins, and low estradiol levels. STAG3 is considered a genetic aetiology of POI, which facilitates entry of REC8 into the nucleus of a cell and plays an essential role in gametogenesis. At present, only six truncated variants associated with POI have been reported; there have been no reports of an in-frame variant of STAG3 causing POI. In this study, two novel homozygous in-frame variants (c.877_885del, p.293_295del; c.891_893dupTGA, p.297_298insAsp) in STAG3 were identified in two sisters with POI from a five-generation consanguineous Han Chinese family. To evaluate the effects of these two variants, we performed fluorescence localization and co-immunoprecipitation analyses using in vitro cell model. The two variants were shown to be pathogenic, as neither STAG3 nor REC8 entered nuclei, and interactions between mutant STAG3 and REC8 or SMC1A were absent. To the best of our knowledge, this is the first report on in-frame variants of STAG3 that cause POI. This finding extends the spectrum of variants in STAG3 and sheds new light on the genetic origins of POI.

Background Interleukin-17 (IL-17), the characteristic cytokine secreted by T helper 17 lymphocytes (Th17 cells), plays a pivotal role in host defense and many inflammatory or autoimmune diseases. The aim of this study was to obtain purified protein caprine IL-17A (cIL-17A) as an antigen for preparing an IL-17A-specific monoclonal antibody (mAb). Results The coding sequence (CDS) region of cIL-17A was cloned from the peripheral blood mononuclear cells (PBMCs) of dairy goats and then inserted into the expression vector PET 32a and transformed into competent TransB (DE3) cells. Recombinant fusion protein obtained under optimized conditions was used to immunize BALB/c mice for preparing monoclonal antibodies. Finally, the supernatants of two hybridoma cell lines showing positive reaction with the recombinant fusion protein and negative reaction with fusion tags of PET 32a were collected for western blot, immunofluorescence (IF) and immunohistochemistry (IHC) analysis. Our results showed that the maximum amount of soluble protein could be obtained directly in the supernatant when the recombinant expression cells were induced by isopropyl-β-d-thiogalactoside (IPTG) at a concentration of 0.3 mmol/L at 16 °C for 42 h. Western blot analysis showed that the mAb H8 could recognize the eukaryotically expressed cIL-17A in the supernatant of transfected HEK293T cells. Immunofluorescence and immunohistochemistry assays showed that mAb H8 could strongly recognize both the eukaryotically expressed and natural cIL-17A. Conclusions The monoclonal antibody mAb H8 prepared in this study may be a potential tool for the detection of cIL-17A and beneficial for investigating the pathogenesis of various IL-17-associated diseases. Electronic supplementary material The online version of this article (10.1186/s12896-019-0543-5) contains supplementary material, which is available to authorized users.

Sepsis remains a significant health care issue in clinical practice due to its high mortality rate and healthcare cost, despite extensive efforts to better understand the pathophysiology of sepsis. The systemic inflammatory response often leads to severe liver injury, even acute liver dysfunction and failure. Acetic acid, as a type of chemical compound, has been reported to be an emerging drug for improving metabolic syndrome and inhibiting inflammation in rats and human. To verify the effects of acetic acid in protecting the liver and reducing the inflammatory response, a septic mouse model was established by cecal ligation and puncture (CLP), and then the CLP-model mice were treated with acetic acid or PBS. Following the treatment, it was determined that, in CLP-model mice, acetic acid could alleviate the inflammatory response by decreasing the expression of cytokines including interleukin-6 and tumor necrosis factor-α. Additionally, acetic acid also alleviated the liver injury, and the levels of alanine aminotransaminase, aspartate aminotransferase, Toll-like receptor (TLR)4 and nuclear factor-κB (NF-κB) were decreased. The expression of tripartite motif-containing protein (TRIM)40 was also upregulated significantly. Therefore, the authors of the current study hypothesized that acetic acid could decrease the inflammatory response by increasing the expression of TRIM40 and TRIM40 may regulate the activity of the TLR4 signaling pathway. To further illustrate the interaction between TRIM40 and the TLR4 signaling pathway, the authors collected macrophages from the peritoneal cavity by intraperitoneally administering mice with 5 ml ice-cold normal saline. Following the collection, peritoneal macrophages were treated with acetic acid, TRIM40 small interfering RNA or PBS. It was demonstrated that acetic acid upregulated the expression of TRIM40. When TRIM40 was silenced, the protective effect of acetic acid would be reversed as well. The results suggested that TRIM40 could act on and downregulate the activity of the TLR4 signaling pathway. TRIM40 is possibly the major target for acetic acid, which may function as a protective factor in septic mice.

strongBACKGROUND/strongAcute respiratory distress syndrome (ARDS), which is characterized by severe hypoxemia (PaO2/FIO2 ≤300 mmHg), is usually companied by uncontrolled inflammation, oxidative injury, and the damage to the alveolar-capillary barrier. Severe ARDS is usually companied with acute lung injury that worsen the patients' condition. HIPK1 is a modulator of homeodomain-containing transcription factors and regulates multiple cellular biological process associated with inflammation and anti-stress responses.strongMATERIAL AND METHODS/strongWe used an LPS-induced mouse acute lung injury (ALI) model to investigate the possible role of HIPK1 in ALI pathophysiology.strongRESULTS/strongWe found the HIPK1 was elevated in ALI model mice while interference of HIPK1 by siRNA attenuated the inflammation and oxidative stress indicators (H2O2, O-2, and NO). Further research found HIPK1 interference enhanced the autophagy.strongCONCLUSIONS/strongDecreased HIPK1 in ALI showed protective effects in attenuating inflammation and oxidative stress and enhancing autophagy, indicating HIPK1 as a possible target in ALI management.

To explore the feasibility of sentinel lymph node (SLN) identification by contrast-enhanced ultrasound (CEUS) in pre-operative breast cancer patients and the value of enhancement patterns for diagnosing lymph node metastases and characterising axillary nodal burden. 110 consecutive breast cancer patients were enrolled. Before the surgery, microbubbles were injected intradermally. The lymphatic drainage pathway was detected to identify the SLNs. Blue dye and indocyanine green (ICG) fluorescence were used to trace SLNs during the operation. The enhancement patterns of SLNs were recorded and compared with the final pathological diagnosis. SLN detection rate was 96.4 % of 110 patients, 134 SLNs were detected in total. The sensitivity, specificity, positive-predictive value, negative-predictive value and accuracy of predicting SLNs metastases by CEUS enhancement patterns were 100 %, 52.0 %, 43.4 %, 100 % and 64.9 %, respectively. No metastatic SLNs were presented as homogeneous enhancement. Low nodal burden with 0–2 SLN metastases in 92.5 % nodes presented as heterogeneous enhancement. No enhancement pattern was proved to be SLN metastases in 100 % patients. CEUS is a feasible approach for SLN identification. CEUS enhancement patterns can be helpful in recognising metastatic SLNs and nodal burden. • CEUS is a feasible approach for SLN identification and characterisation. • The enhancement patterns on CEUS can be helpful in recognising metastasised SLNs. • Homogeneous enhancement pattern has the highest negative-predictive value.

INTRODUCTION The Saccharomyces cerevisiae 2.0 project (Sc2.0) aims to modify the yeast genome with a series of densely spaced designer changes. Both a synthetic yeast chromosome arm (synIXR) and the entirely synthetic chromosome (synIII) function with high fitness in yeast. For designer genome synthesis projects, precise engineering of the physical sequence to match the specified design is important for the systematic evaluation of underlying design principles. Yeast can maintain nuclear chromosomes as rings, occurring by chance at repeated sequences, although the cyclized format is unfavorable in meiosis given the possibility of dicentric chromosome formation from meiotic recombination. Here, we describe the de novo synthesis of synthetic yeast chromosome V (synV) in the “Build-A-Genome China” course, perfectly matching the designer sequence and bearing loxPsym sites, distinguishable watermarks, and all the other features of the synthetic genome. We generated a ring synV derivative with user-specified cyclization coordinates and characterized its performance in mitosis and meiosis. RATIONALE Systematic evaluation of underlying Sc2.0 design principles requires that the final assembled synthetic genome perfectly match the designed sequence. Given the size of yeast chromosomes, synthetic chromosome construction is performed iteratively, and new mutations and unpredictable events may occur during synthesis; even a very small number of unintentional nucleotide changes across the genome could have substantial effects on phenotype. Therefore, precisely matching the physical sequence to the designed sequence is crucial for verification of the design principles in genome synthesis. Ring chromosomes can extend those design principles to provide a model for genomic rearrangement, ring chromosome evolution, and human ring chromosome disorders. RESULTS We chemically synthesized, assembled, and incorporated designer chromosome synV (536,024 base pairs) of S. cerevisiae according to Sc2.0 principles, based on the complete nucleotide sequence of native yeast chromosome V (576,874 base pairs). This work was performed as part of the “Build-A-Genome China” course in Tianjin University. We corrected all mutations found—including duplications, substitutions, and indels—in the initial synV strain by using integrative cotransformation of the precise desired changes and by means of a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)–based method. Altogether, 3331 corrected base pairs were required to match to the designed sequence. We generated a strain that exactly matches all designer sequence changes that displays high fitness under a variety of culture conditions. All corrections were verified with whole-genome sequencing; RNA sequencing revealed only minor changes in gene expression—most notably, decreases in expression of genes relocated near synthetic telomeres as a result of design. We constructed a functional circular synV (ring_synV) derivative in yeast by precisely joining both chromosome ends (telomeres) at specified coordinates. The ring chromosome showed restoration of subtelomeric gene expression levels. The ring_synV strain exhibited fitness comparable with that of the linear synV strain, revealed no change in sporulation frequency, but notably reduced spore viability. In meiosis, heterozygous or homozygous diploid ring_wtV and ring_synV chromosomes behaved similarly, exhibiting substantially higher frequency of the formation of zero-spore tetrads, a type that was not seen in the rod chromosome diploids. Rod synV chromosomes went through meiosis with high spore viability, despite no effort having been made to preserve meiotic competency in the design of synV. CONCLUSION The perfect designer-matched synthetic chromosome V provides strategies to edit sequence variants and correct unpredictable events, such as off-target integration of extra copies of synthetic DNA elsewhere in the genome. We also constructed a ring synthetic chromosome derivative and evaluated its fitness and stability in yeast. Both synV and synVI can be circularized and can power yeast cell growth without affecting fitness when gene content is maintained. These fitness and stability phenotypes of the ring synthetic chromosome in yeast provide a model system with which to probe the mechanism of human ring chromosome disorders. Synthesis, cyclization, and characterization of synV . ( A ) Synthetic chromosome V (synV, 536,024 base pairs) was designed in silico from native chromosome V (wtV, 576,874 base pairs), with extensive genotype modification designed to be phenotypically neutral. ( B ) CRISPR/Cas9 strategy for multiplex repair. ( C ) Colonies of wtV, synV, and ring_synV strains.