Follicular helper T(Tfh)cells select mutated B cells in germinal centres,which can then differentiate into long-lived high affinity memory B cells and plasma cells.Tfh cells are regulated by a unique molecular program...Follicular helper T(Tfh)cells select mutated B cells in germinal centres,which can then differentiate into long-lived high affinity memory B cells and plasma cells.Tfh cells are regulated by a unique molecular programme orchestrated by the transcriptional repressor Bcl6.This transcription factor turns down expression of multiple genes,including transcriptional regulators of other T helper lineages and a vast amount of microRNAs.This enables Tfh cells to express a suite of chemokine receptors,stimulatory ligands and cytokines that enable migration into B-cell follicles,and provision of effective help to B cells.Not surprisingly,dysregulation of this powerful helper subset can lead to a range of autoantibody-mediated diseases;indeed,aberrant accumulation of Tfh cells has been linked with systemic lupus erythematosus,Sjogren’s disease and autoimmune arthritis.Here we dissect multiple checkpoints that operate throughout Tfh cell development and maturation to maintain immunological tolerance while mounting robust and long-lasting antibody responses.展开更多
The Sertoli cell tight junction （T J） is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated clau...The Sertoli cell tight junction （T J） is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine knockout results in infertility. We aimed to assess quantitatively the significance of the contribution of claudin-11 to TJ function, in vitro, using siRNA-mediated gene silencing. We also conducted an analysis of the contribution of occludin, another intrinsic transmembrane protein of the TJ. Silencing of claudin-11 and/or occludin was conducted using siRNA in an immature rat Sertoli cell culture model. Transepithelial electrical resistance was used to assess quantitatively TJ function throughout the culture. Two days after siRNA treatment, cells were fixed for immunocytochemical localization of junction proteins or lyzed for RT-PCR assessment of mRNA expression. Silencing of claudin-11, occludin, or both resulted in significant decreases in TJ function of 55% （P 〈 0.01）, 51% （P 〈 0.01）, and 62% （P 〈 0.01）, respectively. Data were concomitant with significant decreases in mRNA expression and marked reductions in the localization of targeted proteins to the Sertoli cell TJ. We provide quantitative evidence that claudin-11 contributes significantly （P 〈 0.01） to Sertoli cell TJ function in vitro. Interestingly, occludin, which is hormonally regulated but not implicated in infertility until late adulthood, is also a significant （P 〈 0.01） contributor to barrier function. Our data are consistent with in vivo studies that clearly demonstrate a role for these proteins in maintaining normal TJ barrier structure and function.展开更多
AIM: To investigate over-expression of Osteopontin (OPN) pathway expression and mechanisms of action in human alcoholic liver disease (ALD), in vivo and in vitro acute alcohol models.
Background:To generate and validate a method to estimate axial length estimated(AL_(est))from spherical equivalent(SE)and corneal curvature[keratometry(K)],and to determine if this AL_(est)can replace actual axial len...Background:To generate and validate a method to estimate axial length estimated(AL_(est))from spherical equivalent(SE)and corneal curvature[keratometry(K)],and to determine if this AL_(est)can replace actual axial length(AL_(act))for correcting transverse magnification error in optical coherence tomography angiography(OCTA)images using the Littmann-Bennett formula.Methods:Data from 1301 participants of the Raine Study Gen2-20 year follow-up were divided into two datasets to generate(n=650)and validate(n=651)a relationship between AL,SE,and K.The developed formula was then applied to a separate dataset of 46 participants with AL,SE,and K measurements and OCTA images to estimate and compare the performance of AL_(est)against AL_(act)in correcting transverse magnification error in OCTA images when measuring the foveal avascular zone area(FAZA).Results:The formula for AL_(est)yielded the equation:AL_(est)=2.102K−0.4125SE+7.268,R^(2)=0.794.There was good agreement between AL_(est)and AL_(act)for both study cohorts.The mean difference[standard deviation(SD)]between FAZA corrected with AL_(est)and AL_(act)was 0.002(0.015)mm^(2)with the 95%limits of agreement(LoA)of−0.027 to 0.031 mm^(2).In comparison,mean difference(SD)between FAZA uncorrected and corrected with AL_(act)was−0.005(0.030)mm^(2),with 95%LoA of−0.064 to 0.054 mm^(2).Conclusions:AL_(act)is more accurate than AL_(est)and hence should be used preferentially in magnification error correction in the clinical setting.FAZA corrected with AL_(est)is comparable to FAZA corrected with AL_(act),while FAZA measurements using images corrected with AL_(est)have a greater accuracy than measurements on uncorrected images.Hence,in the absence of AL_(act),clinicians should use AL_(est)to correct for magnification error as this provides for more accurate measurements of fundus parameters than uncorrected images.展开更多
Malignant pleural mesothelioma(MPM)is a rare and aggressive cancer with low survival probability as it is generally diagnosed at later stages.'Using a combination of immune-checkpoint inhibitors(ICls)ipilimumab(IP...Malignant pleural mesothelioma(MPM)is a rare and aggressive cancer with low survival probability as it is generally diagnosed at later stages.'Using a combination of immune-checkpoint inhibitors(ICls)ipilimumab(IPl)and nivolumab(NivO)as a first-line treatment for unresectable MPM,the CheckMate 743 trial reported higher overall survival and prolonged duration of response compared with traditional chemotherapy.展开更多
Human pluripotent stem cell(h PSC)-derived progenies are immature versions of cells,presenting a potential limitation to the accurate modelling of diseases associated with maturity or age.Hence,it is important to char...Human pluripotent stem cell(h PSC)-derived progenies are immature versions of cells,presenting a potential limitation to the accurate modelling of diseases associated with maturity or age.Hence,it is important to characterise how closely cells used in culture resemble their native counterparts.In order to select appropriate time points of retinal pigment epithelium(RPE)cultures that reflect native counterparts,we characterised the transcriptomic profiles of the h PSC-derived RPE cells from 1-and 12-month cultures.We differentiated the human embryonic stem cell line H9 into RPE cells,performed single-cell RNA-sequencing of a total of 16,576 cells to assess themolecular changes of the RPE cells across these two culture time points.Our results indicate the stability of the RPE transcriptomic signature,with no evidence of an epithelial–mesenchymal transition,and with the maturing populations of the RPE observed with time in culture.Assessment of Gene Ontology pathways revealed that as the cultures age,RPE cells upregulate expression of genes involved in metal binding and antioxidant functions.This might reflect an increased ability to handle oxidative stress as cells mature.Comparison with native human RPE data confirms a maturing transcriptional profile of RPE cells in culture.These results suggest that long-term in vitro culture of RPE cells allows the modelling of specific phenotypes observed in native mature tissues.Our work highlights the transcriptional landscape of h PSC-derived RPE cells as they age in culture,which provides a reference for native and patient samples to be benchmarked against.展开更多
BACKGROUND The study on predicting the differentiation grade of colorectal cancer(CRC)based on magnetic resonance imaging(MRI)has not been reported yet.Developing a non-invasive model to predict the differentiation gr...BACKGROUND The study on predicting the differentiation grade of colorectal cancer(CRC)based on magnetic resonance imaging(MRI)has not been reported yet.Developing a non-invasive model to predict the differentiation grade of CRC is of great value.AIM To develop and validate machine learning-based models for predicting the differ-entiation grade of CRC based on T2-weighted images(T2WI).METHODS We retrospectively collected the preoperative imaging and clinical data of 315 patients with CRC who underwent surgery from March 2018 to July 2023.Patients were randomly assigned to a training cohort(n=220)or a validation cohort(n=95)at a 7:3 ratio.Lesions were delineated layer by layer on high-resolution T2WI.Least absolute shrinkage and selection operator regression was applied to screen for radiomic features.Radiomics and clinical models were constructed using the multilayer perceptron(MLP)algorithm.These radiomic features and clinically relevant variables(selected based on a significance level of P<0.05 in the training set)were used to construct radiomics-clinical models.The performance of the three models(clinical,radiomic,and radiomic-clinical model)were evaluated using the area under the curve(AUC),calibration curve and decision curve analysis(DCA).RESULTS After feature selection,eight radiomic features were retained from the initial 1781 features to construct the radiomic model.Eight different classifiers,including logistic regression,support vector machine,k-nearest neighbours,random forest,extreme trees,extreme gradient boosting,light gradient boosting machine,and MLP,were used to construct the model,with MLP demonstrating the best diagnostic performance.The AUC of the radiomic-clinical model was 0.862(95%CI:0.796-0.927)in the training cohort and 0.761(95%CI:0.635-0.887)in the validation cohort.The AUC for the radiomic model was 0.796(95%CI:0.723-0.869)in the training cohort and 0.735(95%CI:0.604-0.866)in the validation cohort.The clinical model achieved an AUC of 0.751(95%CI:0.661-0.842)in the training cohort and 展开更多
The major cause of death from breast cancer is not the primary turnout, but relapsing, drug-resistant, metastatic disease. Identifying factors that contribute to aggressive cancer offers important leads for therapy. I...The major cause of death from breast cancer is not the primary turnout, but relapsing, drug-resistant, metastatic disease. Identifying factors that contribute to aggressive cancer offers important leads for therapy. Inherent defence against carcinogens depends on the individual molecular make-up of each person. Important molecular determinants of these responses are under the control of the mouse double minute （MDM） family： comprised of the proteins MDM2 and MDM4. In normal, healthy adult cells, the MDM family functions to critically regulate measured, cellular responses to stress and subsequent recovery. Proper function of the MDM family is vital for normal breast development, but also for preserving genomic fidelity. The MDM family members are best characterized for their negative regulation of the major tumour suppressor p53 to modulate stress responses. Their impact on other cellular regulators is emerging. Inappropriately elevated protein levels of the MDM family are highly associated with an increased risk of cancer incidence. Exploration of the MDM family members as cancer therapeutic targets is relerant for designing tailored anti-cancer treatments, but successful approaches must strategically consider the impact on both the target cancer and adjacent healthy ceils and tissues. This review focuses on recent findings pertaining to the role of the MDM family in normal and malignant breast cells.展开更多
AIM:To investigate the safety and efficacy of adding bevacizumab to first-line chemotherapy in metastatic colorectal cancer patients with peritoneal disease.METHODS:We compared rates of gastrointestinal perforation in...AIM:To investigate the safety and efficacy of adding bevacizumab to first-line chemotherapy in metastatic colorectal cancer patients with peritoneal disease.METHODS:We compared rates of gastrointestinal perforation in patients with metastatic colorectal cancer and peritoneal disease receiving first-line chemotherapy with and without bevacizumab in three distinct cohorts:(1) the AGITG MAX trial(Phase Ⅲ randomised clinical trial comparing capecitabine vs capecitabine and bevacizumab vs capecitabine,bevacizumab and mitomycin C);(2) the prospective Treatment of Recurrent and Advanced Colorectal Cancer(TRACC) registry(any first-line regimen ± bevacizumab);and(3) two cancer centres in New South Wales,Australia [Macarthur Cancer Therapy Centre and Liverpool Cancer Therapy Centre(NSWCC) from January 2005 to Decenber 2012,(any first-line regimen ± bevacizumab).For the AGITG MAX trial capecitabine was compared to the other two arms(capecitabine/bevacizumab and capecitabine/bevacizumab/mitomycin C).In the AGITG MAX trial and the TRACC registry rates of gastrointestinal perforation were also collected in patients who did not have peritoneal metastases.Secondary endpoints included progression-free survival,chemotherapy duration,and overall survival.Time-toevent outcomes were estimated using the Kaplan-Meier method and compared using the log-rank test.RESULTS:Eighty-four MAX,179 TRACC and 69 NSWCC patients had peritoneal disease.There were no gastrointestinal perforations recorded in either the MAX subgroup or the NSWCC cohorts.Of the patients without peritoneal disease in the MAX trial,4/300(1.3%) in the bevacizumab arms had gastrointestinal perforations compared to 1/123(0.8%) in the capecitabine alone arm.In the TRACC registry 3/126(2.4%) patients who had received bevacizumab had a gastrointestinal perforation compared to 1/53(1.9%) in the chemotherapy alone arm.In a further analysis of patients without peritoneal metastases in the TRACC registry,the rate of gastrointestinal perforations was 9/369(2.4%) in the chemother展开更多
The treatment of advanced pancreatic cancer has not moved much beyond single agent gemcitabine until recently when protocols such as FOLFIRINOX(fluorouracil,leucovorin,irinotecan and oxaliplatin)and nab-paclitaxelgemc...The treatment of advanced pancreatic cancer has not moved much beyond single agent gemcitabine until recently when protocols such as FOLFIRINOX(fluorouracil,leucovorin,irinotecan and oxaliplatin)and nab-paclitaxelgemcitabine have demonstrated some improved outcomes.Advances in technology especially in massively parallel genome sequencing has progressed our understanding of the biology of pancreatic cancer especially the candidate signalling pathways that are involved in tumourogenesis and disease course.This has allowed identification of potentially actionable mutations that may be targeted by new biological agents.The heterogeneity of pancreatic cancer makes tumour tissue collection important with the aim of being able to personalise therapies for the individual as opposed to a one size fits all approach to treatment of the condition.This paper reviews the developments in this area of translational research and the ongoing clinical studies that will attempt to move this into the everyday oncology practice.展开更多
Aim:Aberrant microRNA expression is a common event in cancer drug resistance,however its involvement in malignant pleural mesothelioma(MPM)drug resistance is largely unexplored.We aimed to investigate the contribution...Aim:Aberrant microRNA expression is a common event in cancer drug resistance,however its involvement in malignant pleural mesothelioma(MPM)drug resistance is largely unexplored.We aimed to investigate the contribution of microRNAs to the resistance to drugs commonly used in the treatment of MPM.Methods:Drug resistant MPM cell lines were generated by treatment with cisplatin,gemcitabine or vinorelbine.Expression of microRNAs was quantified using RT-qPCR.Apoptosis and drug sensitivity assays were carried out following transfection with microRNA mimics or BCL2 siRNAs combined with drugs.Results:Expression of miR-15a,miR-16 and miR-34a was downregulated in MPM cells with acquired drug resistance.Transfection with miR-15a or miR-16 mimics reversed the resistance to cisplatin,gemcitabine or vinorelbine,whereas miR-34a reversed cisplatin and vinorelbine resistance only.Similarly,in parental cell lines,miR-15a or miR-16 mimics sensitised cells to all drugs,whereas miR-34a increased response to cisplatin and vinorelbine.Increased microRNA expression increased drug-induced apoptosis and caused BCL2 mRNA and protein reduction.RNAi-mediated knockdown of BCL2 partly recapitulated the increase in drug sensitivity in cisplatin and vinorelbine treated cells.Conclusion:Drug-resistant MPM cell lines exhibited reduced expression of tumour suppressor microRNAs.Increasing tumour suppressor of microRNA expression sensitised both drug resistant and parental cell lines to chemotherapeutic agents,in part through targeting of BCL2.Taken together,these data suggest that miR-15a,miR-16 and miR-34a are involved in the acquired and intrinsic drug resistance phenotype of MPM cells.展开更多
Dear Editor,Despite the improved outcome of advanced estrogen receptor-positive(ER+)breast cancer patients treated with endocrine therapy in combination with either a cyclindependent kinase 4/6 inhibitor(CDK4/6i)or a ...Dear Editor,Despite the improved outcome of advanced estrogen receptor-positive(ER+)breast cancer patients treated with endocrine therapy in combination with either a cyclindependent kinase 4/6 inhibitor(CDK4/6i)or a phosphoinositide 3-kinase inhibitor(PI3Ki),the disease will eventually progress,and the optimal treatment strategy upon progression remains undefined[1-4].To address this,we developed MCF-7-and T47D-derived PIK3CAmutated breast cancer cell lines[5]resistant to combined CDK4/6i palbociclib and fulvestrant(MPF-R and TPFR)or combined PI3Ki alpelisib and fulvestrant(MAF-R and TAF-R),respectively(Supplementary Materials and Methods).Drug-sensitive isogenic cells(M-S and T-S)grown in parallel with MPF-R and TPF-R cells and the original MCF-7/S0.5 and T47D cells were analyzed for comparison.展开更多
Overnutrition causes hyperactivation of mTORC1-dependent negative feedback loops leading to the downregulation of insulin signaling and development of insulin resistance.In osteoblasts(OBs),insulin signaling plays a c...Overnutrition causes hyperactivation of mTORC1-dependent negative feedback loops leading to the downregulation of insulin signaling and development of insulin resistance.In osteoblasts(OBs),insulin signaling plays a crucial role in the control of systemic glucose homeostasis.We utilized mice with conditional deletion of Rptor to investigate how the loss of mTORC1 function in OB affects glucose metabolism under normal and overnutrition dietary states.Compared to the controls,chow-fed Rptorob−/−mice had substantially less fat mass and exhibited adipocyte hyperplasia.Remarkably,upon feeding with high-fat diet,mice with pre-and post-natal deletion of Rptor in OBs were protected from diet-induced obesity and exhibited improved glucose metabolism with lower fasting glucose and insulin levels,increased glucose tolerance and insulin sensitivity.This leanness and resistance to weight gain was not attributable to changes in food intake,physical activity or lipid absorption but instead was due to increased energy expenditure and greater whole-body substrate flexibility.RNA-seq revealed an increase in glycolysis and skeletal insulin signaling pathways,which correlated with the potentiation of insulin signaling and increased insulin-dependent glucose uptake in Rptorknockout osteoblasts.Collectively,these findings point to a critical role for the mTORC1 complex in the skeletal regulation of wholebody glucose metabolism and the skeletal development of insulin resistance.展开更多
BACKGROUND Aminoacyl tRNA synthetases/ligases(ARSs)are highly conserved enzymes involved in attaching amino acids to tRNA promoting protein synthesis.Although deficiencies of ARSs localized to the mitochondria classic...BACKGROUND Aminoacyl tRNA synthetases/ligases(ARSs)are highly conserved enzymes involved in attaching amino acids to tRNA promoting protein synthesis.Although deficiencies of ARSs localized to the mitochondria classically present with neuropathology,the clinical features of cytosolic ARS deficiencies are more variable.They have previously been associated with neonatal hepatitis,but never with early-onset inflammatory bowel disease.CASE SUMMARY A nine-year-old Bangladeshi boy presented with neonatal liver failure and deranged clotting,transaminitis and cholestasis.His parents were first cousins.Two older brothers and a sister were well.The patient suffered from loose stools from early infancy which became more troublesome and persistent from five years old with ten bloody motions a day.Repeated endoscopies showed persistent pancolitis,which was refractory to mesalazine,corticosteroids,azathioprine,sirolimus and anti-TNF(adalimumab)therapy,but has improved recently with subcutaneous methotrexate.Whole Genome Sequencing revealed a novel pathogenic missense variant(c.290A>G)in the cytosolic isoleucyl-tRNA synthetase gene,leading to an amino acid substitution(p.Asp97Gly).Pathogenic variants in other genes associated with inflammatory bowel disease(IBD)(ADAM17,EGFR,FOXP3,IL10RA,IL10RB,IL21R,NCF4,STAT3)were excluded.Cytokine assays demonstrated markedly elevated IL-2,IL-5,IL-13,IL-9 and IL-10 by the patient’s CD4+T-cells,while IL-17A,IL-17F,IFNβwere lower,and TNFαnot significantly different when compared to healthy controls.CONCLUSION This case report provides evidence that recessive mutations in cytosolic isoleucyltRNA synthetase are a novel monogenic cause of IBD,which should be considered,particularly in infants and children with a history of neonatal hepatitis and very early-onset IBD poorly responsive to treatment.展开更多
Parkinson's disease (PD) is the most common neurodegenerative movement disorder in the elderly. As the pathogenesis of PD is still not fully understood, medications with the capacity of halting the disease progres...Parkinson's disease (PD) is the most common neurodegenerative movement disorder in the elderly. As the pathogenesis of PD is still not fully understood, medications with the capacity of halting the disease progression are currently unavailable. The discovery of genes that are causative for, or increase susceptibility to PD is pivotal for the development of novel therapeutic approaches, as they are critical for the onset of PD and the molecular pathways underlying its pathogenesis. By reviewing relevant data, we discuss causative genes, and those associated with PD susceptibility and quantitative traits. Through Gene Ontology database and STRING analysis, we emphasize the roles of inorganic cation transmembrane transport pathways and hypothalamic pituitary thyroid axis, in addition to the established roles of inflammation/oxidative stress and mitochondrial dysfunction in the pathogenesis of PD. It is hoped these insights 1) untangle the clinical complex presentations of PD, 2) reveal the interwoven molecular network leading to PD, and 3) identify critical molecular targets to facilitate novel PD drug discovery, with a view to providing improved consultation and personalized medicine for patients with PD in the future.展开更多
Although doping with growth hormone （GH） is banned, there is anecdotal evidence that it is widely abused. GH is reportedly used often in combination with anabolic steroids at high doses for several months. Developme...Although doping with growth hormone （GH） is banned, there is anecdotal evidence that it is widely abused. GH is reportedly used often in combination with anabolic steroids at high doses for several months. Development of a robust test for GH has been challenging because recombinant human 22 kDa （22K） GH used in doping is indistinguishable analytically from endogenous GH and there are wide physiological fluctuations in circulating GH concentrations. One approach to GH testing is based on measurement of different circulating GH isoforms using immunoassays that differentiate between 22K and other GH isoforms. Administration of 22K GH results in a change in its abundance relative to other endogenous pituitary GH isoforms. The differential isoform method has been implemented; however, its utility is limited because of the short window of opportunity of detection. The second approach, which will extend the window of opportunity of detection, is based on the detection of increased levels of circulating GH-responsive proteins, such as insulin-like growth factor （IGF） axis and collagen peptides. Age and gender are the major determinants of variability for IGF-I and the collagen markers; therefore, a test based on these markers must take age into account for men and women. Extensive data is now available that validates the GH- responsive marker approach and implementation is now largely dependent on establishing an assured supply of standardized assays. Future directions will include more widespread implementation of both approaches by the World Anti-Doping Agency, possible use of other platforms for measurement and an athlete＇s passport to establish individual reference levels for biological parameters such as GH-responsive markers. Novel approaches include gene expression and proteomic profiling.展开更多
Novel visualization methods and strategies are necessary to cope with the deluge of datasets present in any scientific field to make discoveries and find answers to previously unanswered questions.These methods and st...Novel visualization methods and strategies are necessary to cope with the deluge of datasets present in any scientific field to make discoveries and find answers to previously unanswered questions.These methods and strategies should not only present scientific findings as images in a concise way but also need to be effective and expressive,which often remain untested.Here,we present Versus,a tool to enable easy image quality assessment and image ranking,utilizing a two-alternative forced choice methodology(2AFC)and an efficient ranking algorithm based on a binary search.The tool provides a systematic way of setting up evaluation experiments via the web without the necessity to install any additional software or require any programming skills.Furthermore,Versus can easily interface with crowdsourcing platforms,such as Amazon’s Mechanical Turk,or can be used as a stand-alone system to carry out evaluations with experts.We demonstrate the use of Versus by means of an image evaluation study,aiming to determine if hue,saturation,brightness,and texture are good indicators of uncertainty in three-dimensional protein structures.Drawing from the power of crowdsourcing,we argue that there is demand and also great potential for this tool to become a standard for simple and fast image evaluations,with the aim to test the effectiveness and expressiveness of scientific visualizations.展开更多
Cardiovascular disease(CVD) is the leading cause of morbidity and mortality among patients with diabetes mellitus,who have a risk of cardiovascular mortality two to four times that of people without diabetes.An indivi...Cardiovascular disease(CVD) is the leading cause of morbidity and mortality among patients with diabetes mellitus,who have a risk of cardiovascular mortality two to four times that of people without diabetes.An individualised approach to cardiovascular risk estimation and management is needed.Over the past decades,many risk scores have been developed to predict CVD.However,few have been externally validated in a diabetic population and limited studies have examined the impact of applying a prediction model in clinical practice.Currently,guidelines are focused on testing for CVD in symptomatic patients.Atypical symptoms or silent ischemia are more common in the diabetic population,and with additional markers of vascular disease such as erectile dysfunction and autonomic neuropathy,these guidelines can be difficult to interpret.We propose an algorithm incorporating cardiovascular risk scores in combination with typical and atypical signs and symptoms to alert clinicians to consider further investigation with provocative testing.The modalities for investigation of CVD are discussed.展开更多
文摘Follicular helper T(Tfh)cells select mutated B cells in germinal centres,which can then differentiate into long-lived high affinity memory B cells and plasma cells.Tfh cells are regulated by a unique molecular programme orchestrated by the transcriptional repressor Bcl6.This transcription factor turns down expression of multiple genes,including transcriptional regulators of other T helper lineages and a vast amount of microRNAs.This enables Tfh cells to express a suite of chemokine receptors,stimulatory ligands and cytokines that enable migration into B-cell follicles,and provision of effective help to B cells.Not surprisingly,dysregulation of this powerful helper subset can lead to a range of autoantibody-mediated diseases;indeed,aberrant accumulation of Tfh cells has been linked with systemic lupus erythematosus,Sjogren’s disease and autoimmune arthritis.Here we dissect multiple checkpoints that operate throughout Tfh cell development and maturation to maintain immunological tolerance while mounting robust and long-lasting antibody responses.
文摘The Sertoli cell tight junction （T J） is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine knockout results in infertility. We aimed to assess quantitatively the significance of the contribution of claudin-11 to TJ function, in vitro, using siRNA-mediated gene silencing. We also conducted an analysis of the contribution of occludin, another intrinsic transmembrane protein of the TJ. Silencing of claudin-11 and/or occludin was conducted using siRNA in an immature rat Sertoli cell culture model. Transepithelial electrical resistance was used to assess quantitatively TJ function throughout the culture. Two days after siRNA treatment, cells were fixed for immunocytochemical localization of junction proteins or lyzed for RT-PCR assessment of mRNA expression. Silencing of claudin-11, occludin, or both resulted in significant decreases in TJ function of 55% （P 〈 0.01）, 51% （P 〈 0.01）, and 62% （P 〈 0.01）, respectively. Data were concomitant with significant decreases in mRNA expression and marked reductions in the localization of targeted proteins to the Sertoli cell TJ. We provide quantitative evidence that claudin-11 contributes significantly （P 〈 0.01） to Sertoli cell TJ function in vitro. Interestingly, occludin, which is hormonally regulated but not implicated in infertility until late adulthood, is also a significant （P 〈 0.01） contributor to barrier function. Our data are consistent with in vivo studies that clearly demonstrate a role for these proteins in maintaining normal TJ barrier structure and function.
基金Supported by Philanthropic Anonymous Sourcethe University of Sydney Bridging Support Grant,in part for Honours ProjectSupported by the National Health and Medical Research Council,No.NHMRC Practitioner Research Fellowship for PH support
文摘AIM: To investigate over-expression of Osteopontin (OPN) pathway expression and mechanisms of action in human alcoholic liver disease (ALD), in vivo and in vitro acute alcohol models.
文摘Background:To generate and validate a method to estimate axial length estimated(AL_(est))from spherical equivalent(SE)and corneal curvature[keratometry(K)],and to determine if this AL_(est)can replace actual axial length(AL_(act))for correcting transverse magnification error in optical coherence tomography angiography(OCTA)images using the Littmann-Bennett formula.Methods:Data from 1301 participants of the Raine Study Gen2-20 year follow-up were divided into two datasets to generate(n=650)and validate(n=651)a relationship between AL,SE,and K.The developed formula was then applied to a separate dataset of 46 participants with AL,SE,and K measurements and OCTA images to estimate and compare the performance of AL_(est)against AL_(act)in correcting transverse magnification error in OCTA images when measuring the foveal avascular zone area(FAZA).Results:The formula for AL_(est)yielded the equation:AL_(est)=2.102K−0.4125SE+7.268,R^(2)=0.794.There was good agreement between AL_(est)and AL_(act)for both study cohorts.The mean difference[standard deviation(SD)]between FAZA corrected with AL_(est)and AL_(act)was 0.002(0.015)mm^(2)with the 95%limits of agreement(LoA)of−0.027 to 0.031 mm^(2).In comparison,mean difference(SD)between FAZA uncorrected and corrected with AL_(act)was−0.005(0.030)mm^(2),with 95%LoA of−0.064 to 0.054 mm^(2).Conclusions:AL_(act)is more accurate than AL_(est)and hence should be used preferentially in magnification error correction in the clinical setting.FAZA corrected with AL_(est)is comparable to FAZA corrected with AL_(act),while FAZA measurements using images corrected with AL_(est)have a greater accuracy than measurements on uncorrected images.Hence,in the absence of AL_(act),clinicians should use AL_(est)to correct for magnification error as this provides for more accurate measurements of fundus parameters than uncorrected images.
基金The NCT04631731 is funded by (i)Western Sydney Local Health District Research and Education Grant 2021, (ii)Bristol Myers Squibb (CA209-6KR),and (iii)BGI ANZ Genetic Service Grant 2021.
文摘Malignant pleural mesothelioma(MPM)is a rare and aggressive cancer with low survival probability as it is generally diagnosed at later stages.'Using a combination of immune-checkpoint inhibitors(ICls)ipilimumab(IPl)and nivolumab(NivO)as a first-line treatment for unresectable MPM,the CheckMate 743 trial reported higher overall survival and prolonged duration of response compared with traditional chemotherapy.
基金a National Health and Medical Research Council(NHMRC-Australia)Practitioner Fellowship(awarded to AWH)Career Development Fellowship(awarded to JEP)+10 种基金Senior Research Fellowship(Grant No.1154389,awarded to AP)an Australian Research Council Future Fellowship(Grant No.FT 140100047,awarded to AP)NHMRC project grants(Grant Nos.1138253 awarded to ELF and AP,as well as 1062820 and 1124812 awarded to SHN)a NHMRC synergy grant(Grant No.1181010 awarded to ELF and AP)grants from the Macular Disease Foundation Australia(awarded to AP,JEPAWH)the Jack Brockhoff Foundation(awarded to GEL)the DHB Foundation(awarded to GEL and AP)the Ophthalmic Research Institute of Australia(awarded to AP and AWH)Stem Cells Australia-the Australian Research Council Special Research Initiative in Stem Cell Science(awarded to SHN,AWH,JEP,and AP)the TMG Family Fund(awarded to AP and GEL)。
文摘Human pluripotent stem cell(h PSC)-derived progenies are immature versions of cells,presenting a potential limitation to the accurate modelling of diseases associated with maturity or age.Hence,it is important to characterise how closely cells used in culture resemble their native counterparts.In order to select appropriate time points of retinal pigment epithelium(RPE)cultures that reflect native counterparts,we characterised the transcriptomic profiles of the h PSC-derived RPE cells from 1-and 12-month cultures.We differentiated the human embryonic stem cell line H9 into RPE cells,performed single-cell RNA-sequencing of a total of 16,576 cells to assess themolecular changes of the RPE cells across these two culture time points.Our results indicate the stability of the RPE transcriptomic signature,with no evidence of an epithelial–mesenchymal transition,and with the maturing populations of the RPE observed with time in culture.Assessment of Gene Ontology pathways revealed that as the cultures age,RPE cells upregulate expression of genes involved in metal binding and antioxidant functions.This might reflect an increased ability to handle oxidative stress as cells mature.Comparison with native human RPE data confirms a maturing transcriptional profile of RPE cells in culture.These results suggest that long-term in vitro culture of RPE cells allows the modelling of specific phenotypes observed in native mature tissues.Our work highlights the transcriptional landscape of h PSC-derived RPE cells as they age in culture,which provides a reference for native and patient samples to be benchmarked against.
基金the Fujian Province Clinical Key Specialty Construction Project,No.2022884Quanzhou Science and Technology Plan Project,No.2021N034S+1 种基金The Youth Research Project of Fujian Provincial Health Commission,No.2022QNA067Malignant Tumor Clinical Medicine Research Center,No.2020N090s.
文摘BACKGROUND The study on predicting the differentiation grade of colorectal cancer(CRC)based on magnetic resonance imaging(MRI)has not been reported yet.Developing a non-invasive model to predict the differentiation grade of CRC is of great value.AIM To develop and validate machine learning-based models for predicting the differ-entiation grade of CRC based on T2-weighted images(T2WI).METHODS We retrospectively collected the preoperative imaging and clinical data of 315 patients with CRC who underwent surgery from March 2018 to July 2023.Patients were randomly assigned to a training cohort(n=220)or a validation cohort(n=95)at a 7:3 ratio.Lesions were delineated layer by layer on high-resolution T2WI.Least absolute shrinkage and selection operator regression was applied to screen for radiomic features.Radiomics and clinical models were constructed using the multilayer perceptron(MLP)algorithm.These radiomic features and clinically relevant variables(selected based on a significance level of P<0.05 in the training set)were used to construct radiomics-clinical models.The performance of the three models(clinical,radiomic,and radiomic-clinical model)were evaluated using the area under the curve(AUC),calibration curve and decision curve analysis(DCA).RESULTS After feature selection,eight radiomic features were retained from the initial 1781 features to construct the radiomic model.Eight different classifiers,including logistic regression,support vector machine,k-nearest neighbours,random forest,extreme trees,extreme gradient boosting,light gradient boosting machine,and MLP,were used to construct the model,with MLP demonstrating the best diagnostic performance.The AUC of the radiomic-clinical model was 0.862(95%CI:0.796-0.927)in the training cohort and 0.761(95%CI:0.635-0.887)in the validation cohort.The AUC for the radiomic model was 0.796(95%CI:0.723-0.869)in the training cohort and 0.735(95%CI:0.604-0.866)in the validation cohort.The clinical model achieved an AUC of 0.751(95%CI:0.661-0.842)in the training cohort and
文摘The major cause of death from breast cancer is not the primary turnout, but relapsing, drug-resistant, metastatic disease. Identifying factors that contribute to aggressive cancer offers important leads for therapy. Inherent defence against carcinogens depends on the individual molecular make-up of each person. Important molecular determinants of these responses are under the control of the mouse double minute （MDM） family： comprised of the proteins MDM2 and MDM4. In normal, healthy adult cells, the MDM family functions to critically regulate measured, cellular responses to stress and subsequent recovery. Proper function of the MDM family is vital for normal breast development, but also for preserving genomic fidelity. The MDM family members are best characterized for their negative regulation of the major tumour suppressor p53 to modulate stress responses. Their impact on other cellular regulators is emerging. Inappropriately elevated protein levels of the MDM family are highly associated with an increased risk of cancer incidence. Exploration of the MDM family members as cancer therapeutic targets is relerant for designing tailored anti-cancer treatments, but successful approaches must strategically consider the impact on both the target cancer and adjacent healthy ceils and tissues. This review focuses on recent findings pertaining to the role of the MDM family in normal and malignant breast cells.
基金the National Health and Medical Research Council Clinical Trials Centre were received from Cancer Australia and the Cancer Institute New South Wales for the AGITG MAX Trial
文摘AIM:To investigate the safety and efficacy of adding bevacizumab to first-line chemotherapy in metastatic colorectal cancer patients with peritoneal disease.METHODS:We compared rates of gastrointestinal perforation in patients with metastatic colorectal cancer and peritoneal disease receiving first-line chemotherapy with and without bevacizumab in three distinct cohorts:(1) the AGITG MAX trial(Phase Ⅲ randomised clinical trial comparing capecitabine vs capecitabine and bevacizumab vs capecitabine,bevacizumab and mitomycin C);(2) the prospective Treatment of Recurrent and Advanced Colorectal Cancer(TRACC) registry(any first-line regimen ± bevacizumab);and(3) two cancer centres in New South Wales,Australia [Macarthur Cancer Therapy Centre and Liverpool Cancer Therapy Centre(NSWCC) from January 2005 to Decenber 2012,(any first-line regimen ± bevacizumab).For the AGITG MAX trial capecitabine was compared to the other two arms(capecitabine/bevacizumab and capecitabine/bevacizumab/mitomycin C).In the AGITG MAX trial and the TRACC registry rates of gastrointestinal perforation were also collected in patients who did not have peritoneal metastases.Secondary endpoints included progression-free survival,chemotherapy duration,and overall survival.Time-toevent outcomes were estimated using the Kaplan-Meier method and compared using the log-rank test.RESULTS:Eighty-four MAX,179 TRACC and 69 NSWCC patients had peritoneal disease.There were no gastrointestinal perforations recorded in either the MAX subgroup or the NSWCC cohorts.Of the patients without peritoneal disease in the MAX trial,4/300(1.3%) in the bevacizumab arms had gastrointestinal perforations compared to 1/123(0.8%) in the capecitabine alone arm.In the TRACC registry 3/126(2.4%) patients who had received bevacizumab had a gastrointestinal perforation compared to 1/53(1.9%) in the chemotherapy alone arm.In a further analysis of patients without peritoneal metastases in the TRACC registry,the rate of gastrointestinal perforations was 9/369(2.4%) in the chemother
基金Supported by NHMRC,Pancare Australia,Sydney Catalyst,Royal Australasian College of Physicians to Chin VTNHMRC Programme Grant to Sjoquist KM
文摘The treatment of advanced pancreatic cancer has not moved much beyond single agent gemcitabine until recently when protocols such as FOLFIRINOX(fluorouracil,leucovorin,irinotecan and oxaliplatin)and nab-paclitaxelgemcitabine have demonstrated some improved outcomes.Advances in technology especially in massively parallel genome sequencing has progressed our understanding of the biology of pancreatic cancer especially the candidate signalling pathways that are involved in tumourogenesis and disease course.This has allowed identification of potentially actionable mutations that may be targeted by new biological agents.The heterogeneity of pancreatic cancer makes tumour tissue collection important with the aim of being able to personalise therapies for the individual as opposed to a one size fits all approach to treatment of the condition.This paper reviews the developments in this area of translational research and the ongoing clinical studies that will attempt to move this into the everyday oncology practice.
基金This work was supported by a Cancer Institute New South Wales Program Grant(van Zandwijk N and Reid G)and a Sydney Catalyst PhD fellowship(Williams M).
文摘Aim:Aberrant microRNA expression is a common event in cancer drug resistance,however its involvement in malignant pleural mesothelioma(MPM)drug resistance is largely unexplored.We aimed to investigate the contribution of microRNAs to the resistance to drugs commonly used in the treatment of MPM.Methods:Drug resistant MPM cell lines were generated by treatment with cisplatin,gemcitabine or vinorelbine.Expression of microRNAs was quantified using RT-qPCR.Apoptosis and drug sensitivity assays were carried out following transfection with microRNA mimics or BCL2 siRNAs combined with drugs.Results:Expression of miR-15a,miR-16 and miR-34a was downregulated in MPM cells with acquired drug resistance.Transfection with miR-15a or miR-16 mimics reversed the resistance to cisplatin,gemcitabine or vinorelbine,whereas miR-34a reversed cisplatin and vinorelbine resistance only.Similarly,in parental cell lines,miR-15a or miR-16 mimics sensitised cells to all drugs,whereas miR-34a increased response to cisplatin and vinorelbine.Increased microRNA expression increased drug-induced apoptosis and caused BCL2 mRNA and protein reduction.RNAi-mediated knockdown of BCL2 partly recapitulated the increase in drug sensitivity in cisplatin and vinorelbine treated cells.Conclusion:Drug-resistant MPM cell lines exhibited reduced expression of tumour suppressor microRNAs.Increasing tumour suppressor of microRNA expression sensitised both drug resistant and parental cell lines to chemotherapeutic agents,in part through targeting of BCL2.Taken together,these data suggest that miR-15a,miR-16 and miR-34a are involved in the acquired and intrinsic drug resistance phenotype of MPM cells.
基金funded in part by grants from the Danish Cancer Society(to Henrik J.Ditzel)Health Insurance“Denmark”(to Henrik J.Ditzel)+1 种基金Academy of Geriatric Cancer Research(AgeCare)(to Henrik J.Ditzel)Pink Tribute(to Henrik J.Ditzel).
文摘Dear Editor,Despite the improved outcome of advanced estrogen receptor-positive(ER+)breast cancer patients treated with endocrine therapy in combination with either a cyclindependent kinase 4/6 inhibitor(CDK4/6i)or a phosphoinositide 3-kinase inhibitor(PI3Ki),the disease will eventually progress,and the optimal treatment strategy upon progression remains undefined[1-4].To address this,we developed MCF-7-and T47D-derived PIK3CAmutated breast cancer cell lines[5]resistant to combined CDK4/6i palbociclib and fulvestrant(MPF-R and TPFR)or combined PI3Ki alpelisib and fulvestrant(MAF-R and TAF-R),respectively(Supplementary Materials and Methods).Drug-sensitive isogenic cells(M-S and T-S)grown in parallel with MPF-R and TPF-R cells and the original MCF-7/S0.5 and T47D cells were analyzed for comparison.
基金the National Health and Medical Research Council of Australia(APP1109207,awarded to ACWZ,PMB,and CGP)Australian Research Council(DP160100454,awarded to ACWZ and PMB)+1 种基金Diabetes Australia Research Program(awarded to ACWZ,SF and SM)an Australia Postgraduate Award(PT).
文摘Overnutrition causes hyperactivation of mTORC1-dependent negative feedback loops leading to the downregulation of insulin signaling and development of insulin resistance.In osteoblasts(OBs),insulin signaling plays a crucial role in the control of systemic glucose homeostasis.We utilized mice with conditional deletion of Rptor to investigate how the loss of mTORC1 function in OB affects glucose metabolism under normal and overnutrition dietary states.Compared to the controls,chow-fed Rptorob−/−mice had substantially less fat mass and exhibited adipocyte hyperplasia.Remarkably,upon feeding with high-fat diet,mice with pre-and post-natal deletion of Rptor in OBs were protected from diet-induced obesity and exhibited improved glucose metabolism with lower fasting glucose and insulin levels,increased glucose tolerance and insulin sensitivity.This leanness and resistance to weight gain was not attributable to changes in food intake,physical activity or lipid absorption but instead was due to increased energy expenditure and greater whole-body substrate flexibility.RNA-seq revealed an increase in glycolysis and skeletal insulin signaling pathways,which correlated with the potentiation of insulin signaling and increased insulin-dependent glucose uptake in Rptorknockout osteoblasts.Collectively,these findings point to a critical role for the mTORC1 complex in the skeletal regulation of wholebody glucose metabolism and the skeletal development of insulin resistance.
文摘BACKGROUND Aminoacyl tRNA synthetases/ligases(ARSs)are highly conserved enzymes involved in attaching amino acids to tRNA promoting protein synthesis.Although deficiencies of ARSs localized to the mitochondria classically present with neuropathology,the clinical features of cytosolic ARS deficiencies are more variable.They have previously been associated with neonatal hepatitis,but never with early-onset inflammatory bowel disease.CASE SUMMARY A nine-year-old Bangladeshi boy presented with neonatal liver failure and deranged clotting,transaminitis and cholestasis.His parents were first cousins.Two older brothers and a sister were well.The patient suffered from loose stools from early infancy which became more troublesome and persistent from five years old with ten bloody motions a day.Repeated endoscopies showed persistent pancolitis,which was refractory to mesalazine,corticosteroids,azathioprine,sirolimus and anti-TNF(adalimumab)therapy,but has improved recently with subcutaneous methotrexate.Whole Genome Sequencing revealed a novel pathogenic missense variant(c.290A>G)in the cytosolic isoleucyl-tRNA synthetase gene,leading to an amino acid substitution(p.Asp97Gly).Pathogenic variants in other genes associated with inflammatory bowel disease(IBD)(ADAM17,EGFR,FOXP3,IL10RA,IL10RB,IL21R,NCF4,STAT3)were excluded.Cytokine assays demonstrated markedly elevated IL-2,IL-5,IL-13,IL-9 and IL-10 by the patient’s CD4+T-cells,while IL-17A,IL-17F,IFNβwere lower,and TNFαnot significantly different when compared to healthy controls.CONCLUSION This case report provides evidence that recessive mutations in cytosolic isoleucyltRNA synthetase are a novel monogenic cause of IBD,which should be considered,particularly in infants and children with a history of neonatal hepatitis and very early-onset IBD poorly responsive to treatment.
基金supported by National Natural Science Foundation of China(No.NSFC 82071417,YH)AC received grant funding from the Australian government.
文摘Parkinson's disease (PD) is the most common neurodegenerative movement disorder in the elderly. As the pathogenesis of PD is still not fully understood, medications with the capacity of halting the disease progression are currently unavailable. The discovery of genes that are causative for, or increase susceptibility to PD is pivotal for the development of novel therapeutic approaches, as they are critical for the onset of PD and the molecular pathways underlying its pathogenesis. By reviewing relevant data, we discuss causative genes, and those associated with PD susceptibility and quantitative traits. Through Gene Ontology database and STRING analysis, we emphasize the roles of inorganic cation transmembrane transport pathways and hypothalamic pituitary thyroid axis, in addition to the established roles of inflammation/oxidative stress and mitochondrial dysfunction in the pathogenesis of PD. It is hoped these insights 1) untangle the clinical complex presentations of PD, 2) reveal the interwoven molecular network leading to PD, and 3) identify critical molecular targets to facilitate novel PD drug discovery, with a view to providing improved consultation and personalized medicine for patients with PD in the future.
文摘Although doping with growth hormone （GH） is banned, there is anecdotal evidence that it is widely abused. GH is reportedly used often in combination with anabolic steroids at high doses for several months. Development of a robust test for GH has been challenging because recombinant human 22 kDa （22K） GH used in doping is indistinguishable analytically from endogenous GH and there are wide physiological fluctuations in circulating GH concentrations. One approach to GH testing is based on measurement of different circulating GH isoforms using immunoassays that differentiate between 22K and other GH isoforms. Administration of 22K GH results in a change in its abundance relative to other endogenous pituitary GH isoforms. The differential isoform method has been implemented; however, its utility is limited because of the short window of opportunity of detection. The second approach, which will extend the window of opportunity of detection, is based on the detection of increased levels of circulating GH-responsive proteins, such as insulin-like growth factor （IGF） axis and collagen peptides. Age and gender are the major determinants of variability for IGF-I and the collagen markers; therefore, a test based on these markers must take age into account for men and women. Extensive data is now available that validates the GH- responsive marker approach and implementation is now largely dependent on establishing an assured supply of standardized assays. Future directions will include more widespread implementation of both approaches by the World Anti-Doping Agency, possible use of other platforms for measurement and an athlete＇s passport to establish individual reference levels for biological parameters such as GH-responsive markers. Novel approaches include gene expression and proteomic profiling.
基金This work was supported by CSIRO’s OCE Science Leader programme and Computational and Simulation Sciences platformpartly by the Australian Research Council under Linkage Project LP140100574。
文摘Novel visualization methods and strategies are necessary to cope with the deluge of datasets present in any scientific field to make discoveries and find answers to previously unanswered questions.These methods and strategies should not only present scientific findings as images in a concise way but also need to be effective and expressive,which often remain untested.Here,we present Versus,a tool to enable easy image quality assessment and image ranking,utilizing a two-alternative forced choice methodology(2AFC)and an efficient ranking algorithm based on a binary search.The tool provides a systematic way of setting up evaluation experiments via the web without the necessity to install any additional software or require any programming skills.Furthermore,Versus can easily interface with crowdsourcing platforms,such as Amazon’s Mechanical Turk,or can be used as a stand-alone system to carry out evaluations with experts.We demonstrate the use of Versus by means of an image evaluation study,aiming to determine if hue,saturation,brightness,and texture are good indicators of uncertainty in three-dimensional protein structures.Drawing from the power of crowdsourcing,we argue that there is demand and also great potential for this tool to become a standard for simple and fast image evaluations,with the aim to test the effectiveness and expressiveness of scientific visualizations.
文摘Cardiovascular disease(CVD) is the leading cause of morbidity and mortality among patients with diabetes mellitus,who have a risk of cardiovascular mortality two to four times that of people without diabetes.An individualised approach to cardiovascular risk estimation and management is needed.Over the past decades,many risk scores have been developed to predict CVD.However,few have been externally validated in a diabetic population and limited studies have examined the impact of applying a prediction model in clinical practice.Currently,guidelines are focused on testing for CVD in symptomatic patients.Atypical symptoms or silent ischemia are more common in the diabetic population,and with additional markers of vascular disease such as erectile dysfunction and autonomic neuropathy,these guidelines can be difficult to interpret.We propose an algorithm incorporating cardiovascular risk scores in combination with typical and atypical signs and symptoms to alert clinicians to consider further investigation with provocative testing.The modalities for investigation of CVD are discussed.
