LncRNA PCGEM1 facilitates cervical cancer progression via miR-642a-5p/KIF5B axis

At present,the role of many long non-coding RNAs(lncRNAs)as tumor suppressors in the formation and development of cervical cancer(CC)has been studied.However,lncRNA prostate cancer gene expression marker 1(PCGEM1),whose high expression not only aggravates ovarian cancer but also can induce tumorigenesis and endometrial cancer progression,has not been studied in CC.The objective of this study was to investigate the expression and the underlying role of PCGEM1 in CC.The relative expression of PCGEM1 in CC cells was detected by real-time PCR.After the suppression of PCGEM1 expression by shRNA,the changes in the proliferation,migration,and invasion capacities were detected via CCK-8 assay,EdU assay,and colony formation assay wound healing assay.Transwell assay and the changes in expressions of epithelial-to-mesenchymal transition(EMT)markers were determined by western blot and immunofluorescence.The interplay among PCGEM1,miR-642a-5p,and kinesin family member 5B(KIF5B)was confirmed by bioinformatics analyses and luciferase reporter assay.Results showed that PCGEM1 expressions were up-regulated within CC cells.Cell viabilities,migration,and invasion were remarkably reduced after the suppression of PCGEM1 expression by shRNA in Hela and SiHa cells.N-cadherin was silenced,but E-cadherin expression was elevated by sh-PCGEM1.Moreover,by sponging miR-642a-5p in CC,PCGEM1 was verified as a competitive endogenous RNA(ceRNA)that modulates KIF5B levels.MiR-642a-5p down-regulation partially rescued sh-PCGEM1’s inhibitory effects on cell proliferation,migration,invasion,and EMT process.In conclusion,the PCGEM1/miR-642a-5p/KIF5B signaling axis might be a novel therapeutic target in CC.This study provides a research basis and new direction for targeted therapy of CC.

Effectiveness of a theory-based tailored mHealth physical activity intervention for women undergoing chemotherapy for breast cancer:A quasi-experimental study

Objectives:This study aimed to explore the effectiveness of the theory-based tailored mHealth physical activity(PA)intervention among patients with breast cancer undergoing chemotherapy.Methods:A quasi-experimental study design was adopted.A total of 60 breast cancer patients were selected from two tertiary hospitals in Shanghai and Hangzhou City from September 2019 to August 2021.According to the admission order,30 patients werefirst included in the control group,followed by 30 patients in the intervention group.A smartphone application(app)named“Breast Care”was developed based on social cognitive theory,self-efficacy theory,and the theory of planned behavior.The app integrated various functions,including information browsing,PA monitoring and feedback,symptom reporting,and social interaction.Patients in the intervention group received three months of personalized online PA guidance in addition to routine care.The control group received routine care.Baseline and post-intervention investigations after three months were conducted in two groups using the Short Form of International Physical Activity Questionnaire,the Hospital Anxiety and Depression Scale,and the Functional Assessment of Cancer TherapydBreast cancer.Results:After three months of intervention,compared to the control group,breast cancer patients in the intervention group showed significant improvements in walking,moderate PA,and overall PA(P<0.05).Compared to the baseline data,breast cancer patients in the intervention group had significant improvements in walking and overall PA after three months(P<0.05),whereas the control group experienced significant declines in walking,moderate PA,and overall PA after three months(P<0.05).There were statistically differences between the two groups in scores for anxiety,overall quality of life,and its dimensions,such as physical well-being,emotional well-being,and additional breast cancer well-being(P<0.05).Conclusions:The theory-based tailored mHealth PA intervention has demonstrated a positive impact on promoting PA behavior change and emotional management among breast cancer patients.The‘Breast Care’app integrated various practical behavior change strategies,offering valuable guidance for personalized remote rehabilitation support for cancer patients.

Immune remodulation in pediatric inherited metabolic liver diseases

Inherited metabolic liver diseases arise from genetic mutations that lead to dis-ruptions in liver metabolic pathways and are predominantly observed in pedia-tric populations.The spectrum of genetic metabolic liver disorders is diverse,encompassing a range of conditions associated with aberrations in iron,copper,carbohydrate,lipid,protein,and amino acid metabolism.Historically,research in the domain of genetic metabolic liver diseases has predominantly concentrated on hepatic parenchymal cell alterations.Nevertheless,emerging studies suggest that inherited metabolic liver diseases exert significant influences on the immune microenvironment,both within the liver and systemically.This review endeavors to encapsulate the immunological features of genetic metabolic liver diseases,aiming to expand the horizons of researchers in this discipline,and to elucidate the underlying pathophysiological mechanisms pertinent to hereditary metabolic liver diseases and to propose innovative therapeutic approaches.

Delivery room resuscitation intensity and associated neonatal outcomes of 24^(+0)-31^(+6) weeks'preterm infants in China:a retrospective cross-sectional study

Background The aim of this study was to review current delivery room(DR)resuscitation intensity in Chinese tertiary neonatal intensive care units and to investigate the association between DR resuscitation intensity and short-term outcomes in preterm infants born at 24+0_31+6 weeks gestation age(GA).Methods This was a retrospective cross-sectional study.The source population was infants born at 24+0_31+6 weeks'GA who were enrolled in the Chinese Neonatal Network 2019 cohort.Eligible infants were categorized into five groups:(1)regular care;(2)oxygen supplementation and/or continuous positive airway pressure(O2/CPAP);(3)mask ventilation;(4)endotracheal intubation;and(5)cardiopulmonary resuscitation(CPR).The association between DR resuscitation and shortterm outcomes was evaluated by inverse propensity score-weighted logistic regression.Results Of 7939 infants included in this cohort,2419(30.5%)received regular care,1994(25.1%)received O,/CPAP,1436(18.1%)received mask ventilation,1769(22.3%)received endotracheal intubation,and 321(4.0%)received CPR in the DR.Advanced maternal age and maternal hypertension correlated with a higher need for resuscitation,and antenatal steroid use tended to be associated with a lower need for resuscitation(P<0.001).Severe brain impairment increased significantly with increasing amounts of resuscitation in DR after adjusting for perinatal factors.Resuscitation strategies vary widely between centers,with over 50%of preterm infants in eight centers requiring higher intensity resuscitation.Conclusions Increased intensity of DR interventions was associated with increased mortality and morbidities in very preterm infants in China.There is wide variation in resuscitative approaches across delivery centers,and ongoing quality improvement to standardize resuscitation practices is needed.

Shanghai Gynecologic Oncology Group’s consensus on the academic and industry’s clinical trial types

Shanghai Gynecologic Oncology Group(SGOG,www.ShanghaiGOG.org),[1]established in 2009,is a non-profit organization of clinical research and the full member of Gynecology Cancer Intergroup(GCIG)since 2012.It was guided by Drs.Jinghe Lang(China),Gavin Stuart(Canada)(2009-2020);Drs.Jinghe Lang(China),Ding Ma(China)(2021-)and was supported by Dr.Zeyi Cao(2009-2012).The mission of SGOG is to actively promote the investigator-initiated trial(IIT)in gynecologic cancers,particularly the innovative phase II clinical trials.Followed by the high quality of management in investigators’training,trial development,human resources.

Gut microbiota and diabetic kidney diseases: Pathogenesis and therapeutic perspectives

Diabetic kidney disease(DKD)is one of the major chronic complications of diabetes mellitus(DM),as well as a main cause of end-stage renal disease.Over the last few years,substantial research studies have revealed a contributory role of gut microbiota in the process of DM and DKD.Metabolites of gut microbiota like lipopolysaccharide,short-chain fatty acids,and trimethylamine N-oxide are key mediators of microbial–host crosstalk.However,the underlying mechanisms of how gut microbiota influences the onset and progression of DKD are relatively unknown.Besides,strategies to remodel the composition of gut microbiota or to reduce the metabolites of microbiota have been found recently,representing a new potential remedial target for DKD.In this minireview,we will address the possible contribution of the gut microbiota in the pathogenesis of DKD and its role as a therapeutic target.

Termination of a partial hydatidiform mole and coexisting fetus: A case report

BACKGROUND We describe the treatment strategy for a patient who was found to have a partial hydatidiform mole and coexisting fetus(PHMCF)during the second trimester.The patient was a 38-year-old Chinese woman who had become pregnant following in vitro fertilization and embryo transplantation.We wanted to determine the safest therapeutic strategy to terminate the PHMCF during the second trimester.CASE SUMMARY In this case,we present a patient who was found to have a PHMCF complicated with serious continuous vaginal bleeding and pre-eclampsia during the second trimester.After careful evaluation,the pregnancy was considered to be unsustainable and was terminated via caesarean section(CS).An infant with weak vital signs and a partially cystic placenta measuring 110 mm×95 mm×35 mm were delivered by CS.The patient was discharged after 4 d.The serum levels ofβ-human chorionic gonadotropin decreased to within a normal range 5 wk after the operation,and no evidence of persistent trophoblastic disease or lung metastases was noticed at the 6-mo follow-up.CONCLUSION CS termination of PHMCF during the second trimester may be a relatively safe therapeutic strategy.

Cultural competence of nurses in Pudong New Area,Shanghai:a mixed-method study

Objective:Cultural competence has gradually attracted attention from many countries,including China.This study was undertaken to determine the cultural competence of registered nurses in Shanghai,China,and to identify the cultural competence among registered nurses in Pudong New Area,Shanghai.Methods:Qualitative interviews were conducted in combination with a quantitative survey.Fifteen clinical nurses were interviewed,and 1088 clinical nurses were recruited for the survey with cultural competence scale for registered nurses,based on the results of the qualitative and quantitative studies.Results:The overall level of cultural competence among registered nurses in Shanghai's Pudong New Area was moderate.Among the seven dimensions,cultural encounter had the highest score,followed by cultural practice,cultural awareness,cultural desire,cultural skill,cultural experience,and cultural knowledge.Age,level of hospital care,mastery of secondary level,and studying overseas were the influencing factors.Conclusions:Hospitals and universities should be aware of the importance of studying cultural competence.Cultural competencerelated courses should be increased,and various forms of training should be undertaken to enhance the interest of nurses.

Development of a surface plasmon resonance biosensor for accurate and sensitive quantitation of small molecules in blood samples

Therapeutic drug monitoring(TDM)has played an important role in clinical medicine for precise dosing.Currently,chromatographic technology and immunoassay detection are widely used in TDM and have met most of the needs of clinical drug therapy.However,some problems still exist in practical applications,such as complicated operation and the influence of endogenous substances.Surface plasmon resonance(SPR)has been applied to detect the concentrations of small molecules,including pesticide residues in crops and antibiotics in milk,which indicates its potential for in vivo drug detection.In this study,a new SPR-based biosensor for detecting chloramphenicol(CAP)in blood samples was developed and validated using methodological verification,including precision,accuracy,matrix effect,and extraction recovery rate,and compared with the classic ultra-performance liquid chromatographyultraviolet(UPLC-UV)method.The detection range of SPR was 0.1-50 ng/mL and the limit of detection was 0.099±0.023 ng/mL,which was lower than that of UPLC-UV.The intra-day and inter-day accuracies of SPR were 98%-114% and 110%-122%,which met the analysis requirement.The results show that the SPR biosensor is identical to UPLC-UV in the detection of CAP in rat blood samples;moreover,the SPR biosensor has better sensitivity.Therefore,the present study shows that SPR technology can be used for the detection of small molecules in the blood samples and has the potential to become a method for therapeutic drug monitoring.

Development of an assessment and intervention protocol for postpartum hemorrhage in the mainland of China: an evidence-based method and Delphi consult

Objective: Postpartum hemorrhage(PPH) is a leading cause of maternal death. Although guidelines have been updated, those with detailed protocols are limited for nursing practice. This study aims at establishing an early assessment and intervention protocol as a toolkit for PPH for midwives and obstetrical nurses.Methods: Employing the evidence-based method, a systematic Internet search of guidelines was conducted and appraisal of literatures was conducted with AGREE system and Oxman-Guyatt Overview Quality Assessment Questionnaire(OQAQ), according to which a protocol draft was therefore developed. Then, a two-round modified Delphi method was utilized to reach a consensus of the protocol built on best practices. Selection criteria for each intervention measure included consensus level with a threshold of 70%, mean of importance(M) >3.5, and coefficient of variation(CV) <0.25. Reliability of experts' opinion was calculated by positive coefficient and authoritative coefficient. Items without consistency were enlisted in the second-round consult. When all items met the selection criteria, the protocol would be finally formulated.Results: A 122-measure protocol was established, including prevention, assessment, and intervention of PPH. With a panel of 14 experts participated in the consult, the positivity coefficient was 0.93 and 1.00 for two rounds, respectively, and the authority coefficient was 0.88. After a two-round consult and revision of the draft, the final program was formulated, containing 5 first-level indexes and 14 second-level indexes with a total item of 120.Conclusions: The PPH protocol, based on high-quality evidences, was formulated with a two-round Delphi method, which can provide insight for midwives and obstetrical nurses to effectively deal with PPH.

Gene-knockout by iSTOP enables rapid reproductive disease modeling and phenotyping in germ cells of the founder generation

Cytosine base editing achieves C·G-to-T·A substitutions and can convert four codons(CAA/CAG/CGA/TGG)into STOP-codons(induction of STOP-codons,iSTOP)to knock out genes with reduced mosaicism.iSTOP enables direct phenotyping in founders’somatic cells,but it remains unknown whether this works in founders’germ cells so as to rapidly reveal novel genes for fertility.Here,we initially establish that iSTOP in mouse zygotes enables functional characterization of known genes in founders’germ cells:Cfap43-iSTOP male founders manifest expected sperm features resembling human“multiple morphological abnormalities of the flagella”syndrome(i.e.,MMAF-like features),while oocytes of Zp3-iSTOP female founders have no zona pellucida.We further illustrate iSTOP’s utility for dissecting the functions of unknown genes with Ccdc183,observing MMAF-like features and male infertility in Ccdc183-iSTOP founders,phenotypes concordant with those of Ccdc183-KO offspring.We ultimately establish that CCDC183 is essential for sperm morphogenesis through regulating the assembly of outer dynein arms and participating in the intra-flagellar transport.Our study demonstrates iSTOP as an efficient tool for direct reproductive disease modeling and phenotyping in germ cells of the founder generation,and rapidly reveals the essentiality of Ccdc183 in fertility,thus providing a time-saving approach for validating genetic defects(like nonsense mutations)for human infertility.

多囊卵巢综合征无排卵的胰岛素信号和雄激素合成的新遗传风险和代谢特征

促排卵是多囊卵巢综合征(PCOS)不孕症的一线治疗方案。卵巢对促排卵治疗的排卵应答差被认为与胰岛素抵抗和高雄激素血症相关。在一个包含1000名PCOS不孕妇女(PCOSAct)的前瞻性队列中,我们开展了一项全外显子联合靶向单核苷酸多态性(SNP)测序以及代谢组学研究。在全基因组水平找出与无排卵显著相关的常见变异和罕见突变,并通过机器学习算法构建排卵预测模型。研究发现,ZNF438基因中标记为rs2994652(p=2.47×10^(-8))的常见变异和REC114基因中的一个罕见功能突变(rs182542888,p=5.79×10^(-6))与促排卵治疗失败显著相关。携带rs2994652 A等位基因和REC114 p.Val101Leu(rs182542888)的PCOS不孕妇女进行促排卵治疗的总排卵率更低(分别为比值比(OR)=1.96,95%置信区间(CI)[1.55~2.49];OR=11.52,95%CI[3.08~43.05]),出现排卵的间隔时间更长(平均56.7天vs.49.0天,p<0.001;78.1天vs.68.6天,p=0.014)。对于rs2994652突变者,L-苯丙氨酸水平升高并与胰岛素抵抗稳态模型(HOMA-IR)指数(r=0.22,p=0.05)和空腹血糖(r=0.33,p=0.003)呈正相关;对于rs182542888突变者,花生四烯酸代谢产物水平下降并与升高的抗苗勒管激素(r=-0.51,p=0.01)和总睾酮(r=-0.71,p=0.02)呈负相关。整合基因变异位点、代谢产物及临床特征的联合预测模型可提高对排卵的预测能力[曲线下面积(AUC)=76.7%]。ZNF438基因的一个常见变异和REC114基因的一个罕见功能突变,以及与二者相关的苯丙氨酸和花生四烯酸代谢物改变,与PCOS女性不孕症的促排卵治疗失败相关。

Coiled-coil domain-containing 38 is required for acrosome biogenesis and fibrous sheath assembly in mice

During spermiogenesis,haploid spermatids undergo dramatic morphological changes to form slender sperm flagella and cap-like acrosomes,which are required for successful fertilization.Severe deformities in flagella cause a male infertility syndrome,multiple morphological abnormalities of the flagella(MMAF),while acrosomal hypoplasia in some cases leads to sub-optimal embryonic developmental potential.However,evidence regarding the occurrence of acrosomal hypoplasia in MMAF is limited.Here,we report the generation of base-edited mice knocked out for coiled-coil domain-containing 38(Ccdc38)via inducing a nonsense mutation and find that the males are infertile.The Ccdc38-KO sperm display acrosomal hypoplasia and typical MMAF phenotypes.We find that the acrosomal membrane is loosely anchored to the nucleus and fibrous sheaths are disorganized in Ccdc38-KO sperm.Further analyses reveal that Ccdc38 knockout causes a decreased level of TEKT3,a protein associated with acrosome biogenesis,in testes and an aberrant distribution of TEKT3 in sperm.We finally show that intracytoplasmic sperm injection overcomes Ccdc38-related infertility.Our study thus reveals a previously unknown role for CCDC38 in acrosome biogenesis and provides additional evidence for the occurrence of acrosomal hypoplasia in MMAF.

Comprehensive transcriptional atlas of human adenomyosis deciphered by the integration of single-cell RNA-sequencing and spatial transcriptomics

Adenomyosis is a poorly understood gynecological disorder lacking effective treatments.Controversy persists regarding“invagination”and“metaplasia”theories.The endometrial-myometrial junction(EMJ)connects the endometrium and myometrium and is important for diagnosing and classifying adenomyosis,but its in-depth study is just beginning.Using single-cell RNA sequencing and spatial profiling,we mapped transcriptional alterations across eutopic endometrium,lesions,and EMJ.Within lesions,we identified unique epithelial(LGR5+)and invasive stromal(PKIB+)subpopulations,along with WFDC1+progenitor cells,supporting a complex interplay between“invagination”and“metaplasia”theories of pathogenesis.Further,we observed endothelial cell heterogeneity and abnormal angiogenic signaling involving vascular endothelial growth factor and angiopoietin pathways.Cell-cell communication differed markedly between ectopic and eutopic endometrium,with aberrant signaling in lesions involving pleiotrophin,TWEAK,and WNT cascades.This study reveals unique stem cell-like and invasive cell subpopulations within adenomyosis lesions identified,dysfunctional signaling,and EMJ abnormalities critical to developing precise diagnostic and therapeutic strategies.

Loss of Tet hydroxymethylase activity causes mouse embryonic stem cell differentiation bias and developmental defects

The TET family is well known for active DNA demethylation and plays important roles in regulating transcription,the epigenome and development.Nevertheless,previous studies using knockdown(KD)or knockout(KO)models to investigate the function of TET have faced challenges in distinguishing its enzymatic and nonenzymatic roles,as well as compensatory effects among TET family members,which has made the understanding of the enzymatic role of TET not accurate enough.To solve this problem,we successfully generated mice catalytically inactive for specific Tet members(Tetm/m).We observed that,compared with the reported KO mice,mutant mice exhibited distinct developmental defects,including growth retardation,sex imbalance,infertility,and perinatal lethality.Notably,Tetm/mmouse embryonic stem cells(mESCs)were successfully established but entered an impaired developmental program,demonstrating extended pluripotency and defects in ectodermal differentiation caused by abnormal DNA methylation.Intriguingly,Tet3,traditionally considered less critical for m ESCs due to its lower expression level,had a significant impact on the global hydroxymethylation,gene expression,and differentiation potential of mESCs.Notably,there were common regulatory regions between Tet1 and Tet3 in pluripotency regulation.In summary,our study provides a more accurate reference for the functional mechanism of Tet hydroxymethylase activity in mouse development and ESC pluripotency regulation.

Transcriptome Dynamics and Cell Dialogs Between Oocytes and Granulosa Cells in Mouse Follicle Development

The development and maturation of follicles is a sophisticated and multistage process.The dynamic gene expression of oocytes and their surrounding somatic cells and the dialogs between these cells are critical to this process.In this study,we accurately classified the oocyte and follicle development into nine stages and profiled the gene expression of mouse oocytes and their surrounding granulosa cells and cumulus cells.The clustering of the transcriptomes showed the trajectories of two distinct development courses of oocytes and their surrounding somatic cells.Gene expression changes precipitously increased at Type 4 stage and drastically dropped afterward within both oocytes and granulosa cells.Moreover,the number of differentially expressed genes between oocytes and granulosa cells dramatically increased at Type 4 stage,most of which persistently passed on to the later stages.Strikingly,cell communications within and between oocytes and granulosa cells became active from Type 4 stage onward.Cell dialogs connected oocytes and granulosa cells in both unidirectional and bidirectional manners.TGFB2/3,TGFBR2/3,INHBA/B,and ACVR1/1B/2B of TGF-βsignaling pathway functioned in the follicle development.NOTCH signaling pathway regulated the development of granulosa cells.Additionally,many maternally DNA methylation-or H3K27me3-imprinted genes remained active in granulosa cells but silent in oocytes during oogenesis.Collectively,Type 4 stage is the key turning point when significant transcription changes diverge the fate of oocytes and granulosa cells,and the cell dialogs become active to assure follicle development.These findings shed new insights on the transcriptome dynamics and cell dialogs facilitating the development and maturation of oocytes and follicles.

miR-125b Confers Resistance of Ovarian Cancer Cells to Cisplatin by Targeting Pro-apoptotic Bcl-2 Antagonist Killer 1

Chemotherapy is the preferred therapeutic approach for advanced ovarian cancer,but a successful long-term treatment is prevented by the development of drug resistance.Recent works have underlined the involvement of non-coding RNAs,microRNAs（miRNAs） in cancer development,with several conjectures regarding their possible involvement in the evolution of drug resistance.This study is to investigate the promoting effects and mechanism of miR-125b involved in the development of chemoresistance in ovarian cancer.The different expression of miR-125b in cisplatin-sensitive ovarian cancer cell line（OV2008） and its resistant variant（C13＊） was identified by real-time PCR.An in vitro cytotoxicity assay and apoptosis assay using CCK-8 assay and flow cytometry,were carried out to detect the effect of miR-125b and Bak1 on cisplatin resistance of cells.Real-time PCR,Western blotting and luciferase reporter assay were used to detect whether Bak1 is a target of miR-125b.As compared with OV2008 cells,the expression levels of miR-125b in C13＊ cells were increased.It was found that the up-regulation of microRNA-125b caused a marked inhibition of cisplatin-induced cytotoxicity and apoptosis and a subsequent increase in the resistance to cisplatin in OV2008 and C13＊ cells.Moreover,Bak1 was a direct target of miR-125b,and down-regulation of Bak1 suppressed cisplatin-induced apoptosis and led to an increased resistance to cisplatin.Our study indicates that miR-125b has a significantly promoting effect on chemoresistance of C13＊ cells and up-regulation of miR-125b expression contributes to cisplatin resistance through suppression of Bak1 expression.This finding has important implications in the development of targeted therapeutics for overcoming cisplatin resistance in ovarian cancer.

Neuronal-like cell differentiation of non-adherent bone marrow cell-derived mesenchymal stem cells

Non-adherent bone marrow cell-derived mesenchymal stem cells from C57BL/6J mice were sepa- rated and cultured using the ＂pour-off＂ method. Non-adherent bone marrow cell-derived mesen- chymal stem ceils developed colony-forming unit-fibroblasts, and could be expanded by supple- mentation with epidermal growth factor. Immunocytochemistry showed that the non-adherent bone marrow cell-derived mesenchymal stem cells exposed to basic fibroblast growth factor/epidermal growth factor/nerve growth factor expressed the neuron specific markers, neurofilament-200 and NeuN, in vitro. Non-adherent bone marrow cell-derived mesenchymal stem cells from 13-galactosidase transgenic mice were also transplanted into focal ischemic brain （right corpus striatum） of C57BL/6J mice. At 8 weeks, cells positive for LacZ and 13-galactosidase staining were observed in the ischemic tissues, and cells co-labeled with both 13-galactosidase and NeuN were seen by double immunohistochemical staining. These findings suggest that the non-adherent bone marrow cell-derived mesenchymal stem cells could differentiate into neuronal-like cells in vitro and in vivo.

Response of gut microbiota to serum metabolome changes in intrahepatic cholestasis of pregnant patients

BACKGROUND Intrahepatic cholestasis in pregnancy(ICP)is the most common liver disease during pregnancy,and its exact etiology and course of progression are still poorly understood.AIM To investigate the link between the gut microbiota and serum metabolome in ICP patients.METHODS In this study,a total of 30 patients were recruited,including 15 patients with ICP(disease group)and 15 healthy pregnant patients(healthy group).The serum nontarget metabolomes from both groups were determined.Amplification of the 16S rRNA V3-V4 region was performed using fecal samples from the disease and healthy groups.By comparing the differences in the microbiota and metabolite compositions between the two groups,the relationship between the gut microbiota and serum metabolites was also investigated.RESULTS The Kyoto Encyclopedia of Genes and Genomes analysis results showed that the primary bile acid biosynthesis,bile secretion and taurine and hypotaurine metabolism pathways were enriched in the ICP patients compared with the healthy controls.In addition,some pathways related to protein metabolism were also enriched in the ICP patients.The principal coordination analysis results showed that there was a distinct difference in the gut microbiota composition(beta diversity)between the ICP patients and healthy controls.At the phylum level,we observed that the relative abundance of Firmicutes was higher in the healthy group,while Bacteroidetes were enriched in the disease group.At the genus level,most of the bacteria depleted in ICP are able to produce short-chain fatty acids(e.g.,Faecalibacterium,Blautia and Eubacterium hallii),while the bacteria enriched in ICP are associated with bile acid metabolism(e.g.,Parabacteroides and Bilophila).Our results also showed that specific genera were associated with the serum metabolome.CONCLUSION Our study showed that the serum metabolome was altered in ICP patients compared to healthy controls,with significant differences in the bile,taurine and hypotaurine metabolite pathways.Alterations in the metabolization of these pathways may lead to disturbances in the gut microbiota,which may further affect the course of progression of ICP.

IDDoR:a novel reporter mouse system for simultaneous and quantitative in vivo analysis of both DNA double-strand break repair pathways

Dear Editor,Two distinct pathways,non-homologous end joining(NHEJ)and homologous recombination(HR)repair,have evolved to repair DNA double-strand breaks(DSBs),the most deleterious type of DNA damage.Impaired NHEJ and HR are often associated with a high incidence of tumorigenesis and the early onset of aging.A number of methods and tools have been developed to help elucidate the regulatory mechanisms of the two repair pathways in the past.In vitro biochemical assays employ 32P-labeled broken DNA fragments and cell extracts to evaluate DSB repair capacity.At DSB sites,several DSB repair factors form foci,which can be visualized through immunofluorescence and microscopy.Among them,phosphorylated H2AX at the Ser139 residue is often considered a DSB marker,and its formation and removal at different time points post the induction of DSBs can be used to reflect the repair capacity and genomic instability.In addition,by quantifying the kinetics of the recruitment of specific NHEJ and HR factors such as 53BP1,DNA-PKcs,MRE11,RPA2,and RAD51,one would be able to make judgments on whether NHEJ,HR,or both pathways is/are affected and sometimes at which steps DSB repair pathways are regulated.An alternative approach for kinetic analysis is to quantify the microirradiation-induced recruitment of repair factors.Moreover,comet assays are also utilized to measure DNA damage-induced genomic stability at the single-cell level.However,with these assays,it is still difficult to measure NHEJ and HR efficiency in a relatively less time-consuming and more quantitative manner.