Use of animal models in preclinical transplant research is essential to the optimization of human allografts for clinical transplantation.Animal models of organ donation and preservation help to advance and improve te...Use of animal models in preclinical transplant research is essential to the optimization of human allografts for clinical transplantation.Animal models of organ donation and preservation help to advance and improve technical elements of solid organ recovery and facilitate research of ischemia-reperfusion injury,organ preservation strategies,and future donor-based interventions.Important considerations include cost,public opinion regarding the conduct of animal research,translational value,and relevance of the animal model for clinical practice.We present an overview of two porcine models of organ donation:donation following brain death(DBD)and donation following circulatory death(DCD).The cardiovascular anatomy and physiology of pigs closely resembles those of humans,making this species the most appropriate for pre-clinical research.Pigs are also considered a potential source of organs for human heart and kidney xenotransplantation.It is imperative to minimize animal loss during procedures that are surgically complex.We present our experience with these models and describe in detail the use cases,procedural approach,challenges,alternatives,and limitations of each model.展开更多
Objective The present study aims to find a convenient, rapid, and stable method to establish bladder tumor in mice. Methods Female Balb/C-nu-nu nude mice （or female T739 mice） were narcotized by sodium pentobarbital...Objective The present study aims to find a convenient, rapid, and stable method to establish bladder tumor in mice. Methods Female Balb/C-nu-nu nude mice （or female T739 mice） were narcotized by sodium pentobarbital at a dosage of 60 mg/ kg. The stylet of the 24# venous retention needles was bent in a 5° to 7° angle at a distance of 15 mm from the needlepoint to form a circle with 2.61 mm to 3.66 mm radius when the stylet is rotated. The pipe casing was lubricated with liquid paraffin, and inserted into the bladder cavity. The drift angle stylet was inserted into the pipe casing slowly, rotated for five times, and then pulled out. A cell 6 suspension （0.1 mL） of approximately lx10 T24 cells （or BTT cells） was then injected imme＆ately. Results A total of 60 T739 mice and 60 Balb/C-nu-nu nude mice were inoculated with BTT cells and T24 cells, respectively. The bladder tumor incidence and the average survival time of the tumor-bearing mice were 100% and （26.69±9.24） d and 100% and （34.59±9.8） d for the T739 mice and Balb/C-nu-nu nude mice, respectively. Conclusions Using the drift angle stylet to injure the mucous membrane of the urinary bladder can establish a stable bladder transplantable tumor model in mice.展开更多
Obesity is associated with an increased risk of mortality from certain types of cancer, including cancer of the breast. Because obesity is associated with multiple risk factors, however, the exact reasons remain uncle...Obesity is associated with an increased risk of mortality from certain types of cancer, including cancer of the breast. Because obesity is associated with multiple risk factors, however, the exact reasons remain unclear. The objective of this study was to determine which of the risk factors associated with obesity are related to enhanced tumor development. The MMTV-PyMT mouse model develops mammary tumors which share numerous characteristics with those of humans. We challenged these mice with a high fat/high carbohydrate, high caloric (HC) diet, and looked for relationships between enhanced primary tumor development and adiposity, various aspects of glucose homeostasis, and metabolic factors. The HC diet enhanced tumor progression in PyMT mice. While mice on the HC diet also developed increased adiposity, hyperglycemia and hyperinsulinemia, none of these risk factors was found to be associated with the observed increases in tumor growth. Rather, we found that while overall, tumor growth was enhanced in HC diet-fed mice compared to those maintained on a regular diet, it was attenuated in individuals by an HC diet-induced increase in metabolic rate and decrease in respiratory exchange ratio. Tumor size in HC diet-fed mice was directly related to p38 phosphorylation and Bcl-2 inhibition, and the degree of vascularization of these tumors was closely and indirectly related to the rate of mouse oxygen consumption. The data suggest that an increase in metabolic rate and oxygen consumption, induced by the introduction of a high caloric diet, has a protective effect against tumor growth by increasing the activity levels of the tumor suppressor p38 and decreasing the activity of the antiapoptoic protein Bcl-2, as well as by reducing hypoxia-induced tumor vascularization.展开更多
Vitamin D deficiency has been associated with a wide range of diseases and multiple forms of cancer including breast, colon, and prostate cancers. Relatively recent work has demonstrated vitamin D to be critical in im...Vitamin D deficiency has been associated with a wide range of diseases and multiple forms of cancer including breast, colon, and prostate cancers. Relatively recent work has demonstrated vitamin D to be critical in immune function and therefore important in inflammatory diseases such as inflammatory bowel disease(IBD). Because vitamin D deficiency or insufficiency is increasingly prevalent around the world, with an estimated 30%-50% of children and adults at risk for vitamin D deficiency worldwide, it could have a significant impact on IBD. Epidemiologic studies suggest that low serum vitamin D levels are a risk factor for IBD and colon cancer, and vitamin D supplementation is associated with decreased colitis disease activity and/or alleviated symptoms. Patients diagnosed with IBD have a higher incidence of colorectal cancer than the general population, which supports the notion that inflammation plays a key role in cancer development and underscores the importance of understanding how vitamin D influences inflammation and its cancer-promoting effects. In addition to human epidemiological data, studies utilizing mouse models of colitis have shown that vitamin D is beneficial in preventing or ameliorating inflammation and clinical disease. The precise role of vitamin D on colitis is unknown; however, vitamin D regulates immune cell trafficking and differentiation, gut barrier function and antimicrobial peptide synthesis, all of which may be protective from IBD and colon cancer. Here we focus on effects of vitamin D on inflammation and inflammation-associated colon cancer and discuss the potential use of vitamin D for protection and treatment of IBD and colon cancer.展开更多
AIM: To develop a hepatocellular carcinoma (HCC) xenograft model for studying hepatitis C virus (HCV) replication in a mice, and antiviral treatment.METHODS: We developed a stable S3-green fluorescence protein (GFP) c...AIM: To develop a hepatocellular carcinoma (HCC) xenograft model for studying hepatitis C virus (HCV) replication in a mice, and antiviral treatment.METHODS: We developed a stable S3-green fluorescence protein (GFP) cell line that replicated the GFP-tagged HCV sub-genomic RNA derived from a highly efficient JFH1 virus. S3-GFP replicon cell line was injected subcutaneously into γ-irradiated SCID mice. We showed that the S3-GFP replicon cell line formed human HCC xenografts in SCID mice. Cells were isolated from subcutaneous tumors and then serially passaged multiple times in SCID mice by culturing in growth medium supplemented with G-418. The mouse-adapted S3-GFP replicon cells were implanted subcutaneously and also into the liver of SCID mice via intrasplenic infusion to study the replication of HCV in the HCC xenografts. The tumor model was validated for antiviral testing after intraperitoneal injection of interferon-α (IFN-α). RESULTS: A highly tumorigenic S3-GFP replicon cell line was developed that formed subcutaneous tumors within 2 wk and diffuse liver metastasis within 4 wk in SCID mice. Replication of HCV in the subcutaneous and liver tumors was confirmed by cell colony assay, detection of the viral RNA by ribonuclease protection assay and real-time quantitative reverse transcription polymerase chain reaction. High-level replication of HCV sub-genomic RNA in the tumor could be visualized by GFP expression using fluorescence microscopy. IFN-α cleared HCV RNA replication in the subcutaneous tumors within 2 wk and 4 wk in the liver tumor model. CONCLUSION: A non-infectious mouse model allows us to study replication of HCV in subcutaneous and metastatic liver tumors. Clearance of HCV by IFN-α supports use of this model to test other anti-HCV drugs.展开更多
Prion diseases are infectious protein misfolding disorders of the central nervous system that result from misfolding of the cellular prion protein(PrPC)into the pathologic isoform PrPSc.Pathologic hallmarks of prion d...Prion diseases are infectious protein misfolding disorders of the central nervous system that result from misfolding of the cellular prion protein(PrPC)into the pathologic isoform PrPSc.Pathologic hallmarks of prion disease are depositions of pathological prion protein PrPSc,neuronal loss,spongiform degeneration and astrogliosis in the brain.Prion diseases affect human and animals,there is no effective therapy,and they invariably remain fatal.For a long time,neuronal loss was considered the sole reason for neurodegeneration in prion pathogenesis,and the contribution of non-neuronal cells like microglia and astrocytes was considered less important.Recent evidence suggests that neurodegeneration during prion pathogenesis is a consequence of a complex interplay between neuronal and non-neuronal cells in the brain,but the exact role of these non-neuronal cells during prion pathology is still elusive.Astrocytes are non-neuronal cells that regulate brain homeostasis under physiological conditions.However,astrocytes can deposit PrPSc aggregates and propagate prions in prion-infected brains.Additionally,sub-populations of reactive astrocytes that include neurotrophic and neurotoxic species have been identified,differentially expressed in the brain during prion infection.Revealing the exact role of astrocytes in prion disease is hampered by the lack of in vitro models of prion-infected astrocytes.Recently,we established a murine astrocyte cell line persistently infected with mouse-adapted prions,and showed how such astrocytes differentially process various prion strains.Considering the complexity of the role of astrocytes in prion pathogenesis,we need more in vitro and in vivo models for exploring the contribution of sub-populations of reactive astrocytes,their differential regulation of signaling cascades,and the interaction with neurons and microglia during prion pathogenesis.This will help to establish novel in vivo models and define new therapeutic targets against prion diseases.In this review,we will discuss the complex role of astrocytes in prion disease,the existing experimental resources,the challenges to analyze the contribution of astrocytes in prion disease pathogenesis,and future strategies to improve the understanding of their role in prion disease.展开更多
Background:Tiletamine/zolazepam is a dissociative anesthetic combination commonly used in small animals but information is limited in rats.The alpha-2 agonist,dexmedetomidine,has gained popularity in laboratory animal...Background:Tiletamine/zolazepam is a dissociative anesthetic combination commonly used in small animals but information is limited in rats.The alpha-2 agonist,dexmedetomidine,has gained popularity in laboratory animal anesthesia.Tramadol is a weak opioid mu agonist.The aim of this study was to assess whether the tiletamine/zolazepam/dexmedetomidine(ZD)combination effectively provides a surgical anesthesia plane comparable to tiletamine/zolazepam/dexmedetomidine with tramadol(ZDT)in a minor procedure in rats.Methods:Rats were induced with ZD or ZDT.After the loss of paw withdrawal,a small incision was made on the rats’left thighs as a surgical stimulus.Rats were maintained under a surgical anesthesia plane by assessing the loss of the paw withdrawal reflex for 45 minutes,then atipamezole was administered.Monitored anesthesia parameters included:(a)physiological parameters-pulse rate(PR),respiratory rate(RR),tissue oxygen saturation(%SpO 2),and body temperature;(b)duration parameters-induction time,onset and duration of surgical anesthesia plane,onset of recovery,and recovery time.Results:PR was significantly lower at 10 minutes in ZD and 5 minutes in ZDT groups.No difference was observed for RR,%SpO 2,and body temperature.Likewise,there were no differences for duration parameters:induction time was less than 3 minutes;onset and duration of surgical anesthesia plane were approximately 5 and 45 minutes,respectively;onset of recovery(time to move)was 51 minutes;and recovery time was 52 minutes,respectively.Conclusion:These data suggest the ZD combination provides a surgical anesthesia plane comparable to ZDT in a rat incisional pain model.展开更多
Resource-scarce regions with serious COVID-19 outbreaks do not have enough ventilators to support critically ill patients,and these shortages are especially devastating in developing countries.To help alleviate this s...Resource-scarce regions with serious COVID-19 outbreaks do not have enough ventilators to support critically ill patients,and these shortages are especially devastating in developing countries.To help alleviate this strain,we have designed and tested the accessible low-barrier in vivo-validated economical ventilator(ALIVE Vent),a COVID-19-inspired,cost-effective,open-source,in vivo-validated solution made from commercially available components.The ALIVE Vent operates using compressed oxygen and air to drive inspiration,while two solenoid valves ensure one-way flow and precise cycle timing.The device was functionally tested and profiled using a variable resistance and compliance artificial lung and validated in anesthetized large animals.Our functional test results revealed its effective operation under a wide variety of ventilation conditions defined by the American Association of Respiratory Care guidelines for ventilator stockpiling.The large animal test showed that our ventilator performed similarly if not better than a standard ventilator in maintaining optimal ventilation status.The FiO2,respiratory rate,inspiratory to expiratory time ratio,positive-end expiratory pressure,and peak inspiratory pressure were successfully maintained within normal,clinically validated ranges,and the animals were recovered without any complications.In regions with limited access to ventilators,the ALIVE Vent can help alleviate shortages,and we have ensured that all used materials are publicly available.While this pandemic has elucidated enormous global inequalities in healthcare,innovative,cost-effective solutions aimed at reducing socio-economic barriers,such as the ALIVE Vent,can help enable access to prompt healthcare and life saving technology on a global scale and beyond COVID-19.展开更多
Background:Extended-release buprenorphine(XR)is indicated for pain management in rodents,but little is known about its use in mice.This study aimed to investigate whether high dose XR effectively attenuates post-opera...Background:Extended-release buprenorphine(XR)is indicated for pain management in rodents,but little is known about its use in mice.This study aimed to investigate whether high dose XR effectively attenuates post-operative hypersensitivity better than low dose XR in a mouse model of incisional pain.Methods:Mice(n 44)were randomly assigned to 1 of 4 treatment groups:(a)saline(1 ml/kg SC,once);(b)sustained release buprenorphine(Bup-SR,1 mg/kg SC,once);(c)low dose extended-release buprenorphine(XR-lo,3.25 mg/kg SC,once);(d)high dose extended-release buprenorphine(XR-hi,6.5 mg/kg SC,once).On days1,0(4 hours),1,2,and 3,mechanical and thermal hypersensitivities were evaluated,and plasma buprenorphine concentrations were measured.Results:Mechanical(days 0-2)and thermal(days 0-1)hypersensitivities were ob-served in the saline group.Bup-SR,XR-lo,and XR-hi attenuated mechanical hyper-sensitivity on days 0,1,and 2.None of the treatment groups,except XR-Lo on day 0,attenuated thermal hypersensitivity on days 0 or 1.Plasma buprenorphine concen-tration peaked at 4 hours(day 0)in all treatment groups and remained greater than 1 ng/mL on days 0-2.No abnormal clinical observations or gross pathologic findings were seen in any groups.Conclusion:The results indicate XR-hi did not effectively attenuate post-operative hypersensitivity better than XR-lo.Thus both 3.25 and 6.5 mg/kg XR are recom-mended for attenuating post-operative hypersensitivity for at least up to 48 hours in mice.展开更多
Background and Aim: The porcine heart bears the best resemblance to the human heart and remains the preferred preclinical model for anatomical, physiological, and medical device studies. In an effort to study phenomen...Background and Aim: The porcine heart bears the best resemblance to the human heart and remains the preferred preclinical model for anatomical, physiological, and medical device studies. In an effort to study phenomena related strictly to ischemia reperfusion and donor preservation protocols, it is essential to avoid the immune responses related to allotransplantation. Orthotopic auto-transplantation is a unique strategy to the field of cardiac transplantation for ex vivo experimentation. Nevertheless, auto-transplantation carries its own technical challenges related to insufficient length of the great vessels that are to be transected and re-anastomosed. Methods: A novel method for orthotopic cardiac auto-transplantation in the porcine model was developed and was described herein. Porcine models were used for ex vivo experimentation of a novel device to study ischemia reperfusion injury. Results: A total of five porcine models were used for ex vivo experimentation of a novel device to mitigate ischemia reperfusion injury and determine effects of donor preservation. Modifications to routine cardiac transplantation protocols to allow for successful auto-transplantation are described. Conclusion: Orthotopic cardiac auto-transplantation in the porcine model is a plausible and technically feasible method for reliable study of ischemia reperfusion injury and donor preservation protocols. Here, we describe methods for both direct orthotopic porcine cardiac auto-transplantations as well as a simplified protocol that can be substituted for full surgical auto-transplantation for the studies of preservation of donor hearts.展开更多
Carbon monoxide(CO) is an endogenous therapeutic gas with an anti-tumor effect. The precise delivery and controlled release of CO in tumor tissues play crucial roles in anti-cancer treatment. However, efficient in sit...Carbon monoxide(CO) is an endogenous therapeutic gas with an anti-tumor effect. The precise delivery and controlled release of CO in tumor tissues play crucial roles in anti-cancer treatment. However, efficient in situ generation of CO from metal-free COreleasing molecules(CORMs) remains a formidable challenge. Herein, we develop ultrasound(US)-driven self-decomposition porphyrin as organic and metal-free US-CORMs, which can spatiotemporally control the CO release(347 mmol CO/mol porphyrin) efficiently under physiologically harmless US conditions(1.0 MHz, 1.5 W/cm^(2), 50% duty cycle, 50 min). Moreover,porphyrin as a sonosensitizer can also generate reactive oxygen species(ROS) under US treatment to achieve sonodynamic therapy(SDT). Advanced functions of such porphyrin-based CORMs in CO gas-sonodynamic synergistic treatment have been demonstrated by evaluating the in vitro and in vivo anti-tumor effects.展开更多
In a recent publication in Cell,Li et al.reported that inhibiting the spliceosome in human pluripotent stem cells(hPSCs)can induce a stable totipotent state.1 This study signifies the importance of post-transcriptiona...In a recent publication in Cell,Li et al.reported that inhibiting the spliceosome in human pluripotent stem cells(hPSCs)can induce a stable totipotent state.1 This study signifies the importance of post-transcriptional regulation in controlling developmental potential and provides a powerful model system for studying the process of human pre-implantation development in vitro.展开更多
Background： Contribution of model for end-stage liver disease incorporating with serum sodium （MELD-Na） score in predicting acute kidney injury （AKI） after orthotopic liver transplantation （OLT） is yet to be id...Background： Contribution of model for end-stage liver disease incorporating with serum sodium （MELD-Na） score in predicting acute kidney injury （AKI） after orthotopic liver transplantation （OLT） is yet to be identified. This study assessed the prognostic value ot MELD-Na score for the development of AKl following OLT. Methods： Preoperative and surgery-related variables of 321 adult end-stage liver disease patients who underwent OLT in Fuzhou General Hospital were collected. PostoperativeAKI was defined and staged in accordance with the clinical practice guidelines developed by Kidney Disease： Improving Global Outcomes. Univariate and multivariate analysis was performed to determine the risk factors fnr AKI following OLT. The discriminating power of MELD/MELD-Na score on AKI outcome was evaluated by receiver operating characteristic （ROC） curve. Spearman＇s correlation analysis was used for identifying the correlated relationship between MELD/MELD-Na score and the severity levels of AKI. Results： The prevalence of AKI following OLT was in 206 out of 321 patients （64.2%）. Three risk lhctors for AKI post-OLT were presented, preoperative calculated MELD score （odds ratio [OR] = 1.048, P = 0.021）, intraoperative volume of red cell suspension transfusion （OR = 1.001, P 0.002）, and preoperative liver cirrhosis （OR = 2.015, P = 0.012）. Two areas under ROC curve （AUCs） of MELD/MELD-Na score predicting AKI were 0.688 and 0.672, respectively; the difference between two AUCs was not significant （Z= 1.952, P = 0.051）. The Spearman＇s correlation coefficients between MELD/MELD-Na score and the severity levels of AKI were 0.406 and 0.385 （P 0.001, 0.001）, respectively. Conclusions： We demonstrated that preoperative MELD score, intraoperative volume of red cell suspension transfusion and preoperative liver cirrhosis were risk factors for AKI following OLT. Furthermore, we preliminarily validated that MELD score seemed to have a stronger power discriminating AKI post-OLT than that of novel MELD-Na score.展开更多
Uncontrolled bleeding and infection can cause significant increases in mortalities.Hydrogel sealants have attracted extensive attention for their ability to control bleeding.However,because interfacial water is a form...Uncontrolled bleeding and infection can cause significant increases in mortalities.Hydrogel sealants have attracted extensive attention for their ability to control bleeding.However,because interfacial water is a formidable barrier to strong surface bonding,a challenge remains in finding a product that offers robust tissue adhesion combined with anti-infection properties.Inspired by the strong adhesive mechanism of biofilm and mussels,we report a novel dual bionic adhesive hydrogel(DBAH)based on chitosan grafted with methacrylate(CS-MA),dopamine(DA),and N-hydroxymethyl acrylamide(NMA)via a facile radical polymerization process.CS-MA and DA were simultaneously included in the adhesive polymer for imitating the two key adhesive components:polysaccharide intercellular adhesin(PIA)of staphylococci biofilm and 3,4-dihydroxy-L-phenylalanine(Dopa)of mussel foot protein,respectively.DBAH presented strong adhesion at 34 kPa even upon three cycles of full immersion in water and was able to withstand up to 168 mm Hg blood pressure,which is significantly higher than the 60–160 mm Hg measured in most clinical settings.Most importantly,these hydrogels presented outstanding hemostatic capability under wet and dynamic in vivo movements while displaying excellent antibacterial properties and biocompatibility.Therefore,DBAH represents a promising class of biomaterials for high-efficiency hemostasis and wound healing.展开更多
O-linked N-acetyl-glucosamine glycosylation(O-GlcNAcylation)of intracellular proteins is a dynamic process broadly implicated in age-related disease,yet it remains uncharacterized whether and how O-GlcNAcylation contr...O-linked N-acetyl-glucosamine glycosylation(O-GlcNAcylation)of intracellular proteins is a dynamic process broadly implicated in age-related disease,yet it remains uncharacterized whether and how O-GlcNAcylation contributes to the natural aging process.O-GlcNAc transferase(OGT)and the opposing enzyme O-GlcNAcase(OGA)control this nutrient-sensing protein modification in cells.Here,we show that global O-GlcNAc levels are increased in multiple tissues of aged mice.In aged liver,carbamoyl phosphate synthetase 1(CPS1)is among the most heavilyO-GlcNAcylated proteins.CPS1O-GlcNAcylation is reversed by calorie restriction and is sensitive to genetic and pharmacological manipulations of theO-GlcNAc pathway.High glucose stimulates CPS1O-GlcNAcylation and inhibits CPS1 activity.Liver-specific deletion of OGT potentiates CPS1 activity and renders CPS1 irresponsive to further stimulation by a prolonged fasting.Our results identify CPS1 O-GlcNAcylation as a key nutrient-sensing regulatory step in the urea cycle during aging and dietary restriction,implying a role for mitochondrial O-GlcNAcylation in nutritional regulation of longevity.展开更多
Objective:Herbal medicine is an important therapeutic option for benign prostatic hyperplasia(BPH),a common disease in older men that can seriously affect their quality of life.Currently,it is crucial to develop agent...Objective:Herbal medicine is an important therapeutic option for benign prostatic hyperplasia(BPH),a common disease in older men that can seriously affect their quality of life.Currently,it is crucial to develop agents with strong efficacy and few side effects.Herein we investigated the effects of the extract of Rauwolfia vomitoria,a shrub grown in West Africa,on BPH.Methods:Rats with testosterone-induced BPH were treated with R.vomitoria.Prostates were histologically analyzed by Hematoxylin and eosin staining.Proliferation index and the expression levels of androgen receptor and its associated proteins were quantified through immunohistochemistry and immunoblotting.Androgen receptor target genes were examined by quantitative real-time polymerase chain reaction.The sperm count and body weight of rats were also measured.Results:The oral administration of R.vomitoria extract significantly reduced the prostate weight and prostate weight index in BPH rats,supported by the decreased thickness of the prostate epithelial layer and increased lumen size.Similar effects were observed in the BPH rats treated with the reference drug,finasteride.R.vomitoria extract significantly reduced the testosterone-induced proliferation markers,including proliferating cell nuclear antigen and cyclin D1,in the prostate glands of BPH rats;it also reduced levels of androgen receptor,its associated protein steroid 5 a-reductase 1 and its downstream target genes(FK506-binding protein 5 and matrix metalloproteinase 2).Notably,compared with the finasteride group,R.vomitoria extract did not significantly reduce sperm count.Conclusion:R.vomitoria suppresses testosterone-induced BPH development.Due to its milder side effects,R.vomitoria could be a promising therapeutic agent for BPH.展开更多
How distinct transcriptional programs are enacted to generate cellular heterogeneity and plasticity,and enable complex fate decisions are important open questions.One key regulator is the cell’s epigenome state that ...How distinct transcriptional programs are enacted to generate cellular heterogeneity and plasticity,and enable complex fate decisions are important open questions.One key regulator is the cell’s epigenome state that drives distinct transcriptional programs by regulating chromatin accessibility.Genome-wide chromatin accessibility measurements can impart insights into regulatory sequences(in)accessible to DNA-binding proteins at a single-cell resolution.This review outlines molecular methods and bioinformatic tools for capturing cell-to-cell chromatin variation using single-cell assay for transposase-accessible chromatin using sequencing(scATAC-seq)in a scalable fashion.It also covers joint profiling of chromatin with transcriptome/proteome measurements,computational strategies to integrate multi-omic measurements,and predictive bioinformatic tools to infer chromatin accessibility from single-cell transcriptomic datasets.Methodological refinements that increase power for cell discovery through robust chromatin coverage and integrate measurements from multiple modalities will further expand our understanding of gene regulation during homeostasis and disease.展开更多
Sperm-specific phospholipase C zeta(PLCζ)initiates intracellular calcium(Ca2+)transients which drive a series of concurrent events collectively termed oocyte activation.Numerous investigations have linked abrogation ...Sperm-specific phospholipase C zeta(PLCζ)initiates intracellular calcium(Ca2+)transients which drive a series of concurrent events collectively termed oocyte activation.Numerous investigations have linked abrogation and absence/reduction of PLCζwith forms of male infertility in humans where oocyte activation fails.However,very few studies have examined potential relationships between PLCζand advancing male age,both of which are increasingly considered to be major effectors of male fertility.Initial efforts in humans may be hindered by inherent PLCζvariability within the human population,alongside a lack of sufficient controllable repeats.Herein,utilizing immunoblotting,immunofluorescence,and quantitative reverse transcription PCR(qRT-PCR)we examined for the first time PLCζprotein levels and localization patterns in sperm,and PLCζmRNA levels within testes,from mice at 8 weeks,12 weeks,24 weeks,and 36 weeks of age,from two separate strains of mice,C57BL/6(B6;inbred)and CD1(outbred).Collectively,advancing male age generally diminished levels and variability of PLCζprotein and mRNA in sperm and testes,respectively,when both strains were examined.Furthermore,advancing male age altered the predominant pattern of PLCζlocalization in mouse sperm,with younger mice exhibiting predominantly post-acrosomal,and older mice exhibiting both post-acrosomal and acrosomal populations of PLCζ.However,the specific pattern of such decline in levels of protein and mRNA was strain-specific.Collectively,our results demonstrate a negative relationship between advancing male age and PLCζlevels and localization patterns,indicating that aging male mice from different strains may serve as useful models to investigate PLCζin cases of male infertility and subfertility in humans.展开更多
A recent study published in Science by Shrock et al.1 examined how responses to COVID-19 severity differed amongst patients with different prior viral exposure history,finding notable correlations between both.Such se...A recent study published in Science by Shrock et al.1 examined how responses to COVID-19 severity differed amongst patients with different prior viral exposure history,finding notable correlations between both.Such serological profiling provides a window into differential viral responses amongst patients with diverse outcomes,potentially mediating improved therapeutics and vaccines for SARS-CoV-2.展开更多
基金University of Nebraska Collaborative Initiative,Grant/Award Number:Team Seed Grant#26685。
文摘Use of animal models in preclinical transplant research is essential to the optimization of human allografts for clinical transplantation.Animal models of organ donation and preservation help to advance and improve technical elements of solid organ recovery and facilitate research of ischemia-reperfusion injury,organ preservation strategies,and future donor-based interventions.Important considerations include cost,public opinion regarding the conduct of animal research,translational value,and relevance of the animal model for clinical practice.We present an overview of two porcine models of organ donation:donation following brain death(DBD)and donation following circulatory death(DCD).The cardiovascular anatomy and physiology of pigs closely resembles those of humans,making this species the most appropriate for pre-clinical research.Pigs are also considered a potential source of organs for human heart and kidney xenotransplantation.It is imperative to minimize animal loss during procedures that are surgically complex.We present our experience with these models and describe in detail the use cases,procedural approach,challenges,alternatives,and limitations of each model.
基金supported by a grant from the Shanxi Science and Technology Department,China (Noo2010K01)
文摘Objective The present study aims to find a convenient, rapid, and stable method to establish bladder tumor in mice. Methods Female Balb/C-nu-nu nude mice （or female T739 mice） were narcotized by sodium pentobarbital at a dosage of 60 mg/ kg. The stylet of the 24# venous retention needles was bent in a 5° to 7° angle at a distance of 15 mm from the needlepoint to form a circle with 2.61 mm to 3.66 mm radius when the stylet is rotated. The pipe casing was lubricated with liquid paraffin, and inserted into the bladder cavity. The drift angle stylet was inserted into the pipe casing slowly, rotated for five times, and then pulled out. A cell 6 suspension （0.1 mL） of approximately lx10 T24 cells （or BTT cells） was then injected imme＆ately. Results A total of 60 T739 mice and 60 Balb/C-nu-nu nude mice were inoculated with BTT cells and T24 cells, respectively. The bladder tumor incidence and the average survival time of the tumor-bearing mice were 100% and （26.69±9.24） d and 100% and （34.59±9.8） d for the T739 mice and Balb/C-nu-nu nude mice, respectively. Conclusions Using the drift angle stylet to injure the mucous membrane of the urinary bladder can establish a stable bladder transplantable tumor model in mice.
文摘Obesity is associated with an increased risk of mortality from certain types of cancer, including cancer of the breast. Because obesity is associated with multiple risk factors, however, the exact reasons remain unclear. The objective of this study was to determine which of the risk factors associated with obesity are related to enhanced tumor development. The MMTV-PyMT mouse model develops mammary tumors which share numerous characteristics with those of humans. We challenged these mice with a high fat/high carbohydrate, high caloric (HC) diet, and looked for relationships between enhanced primary tumor development and adiposity, various aspects of glucose homeostasis, and metabolic factors. The HC diet enhanced tumor progression in PyMT mice. While mice on the HC diet also developed increased adiposity, hyperglycemia and hyperinsulinemia, none of these risk factors was found to be associated with the observed increases in tumor growth. Rather, we found that while overall, tumor growth was enhanced in HC diet-fed mice compared to those maintained on a regular diet, it was attenuated in individuals by an HC diet-induced increase in metabolic rate and decrease in respiratory exchange ratio. Tumor size in HC diet-fed mice was directly related to p38 phosphorylation and Bcl-2 inhibition, and the degree of vascularization of these tumors was closely and indirectly related to the rate of mouse oxygen consumption. The data suggest that an increase in metabolic rate and oxygen consumption, induced by the introduction of a high caloric diet, has a protective effect against tumor growth by increasing the activity levels of the tumor suppressor p38 and decreasing the activity of the antiapoptoic protein Bcl-2, as well as by reducing hypoxia-induced tumor vascularization.
基金Supported by Grant No.AICR 09A136-RevNIH R21 CA149995-01A1 and NIH 5T32DK007742-17
文摘Vitamin D deficiency has been associated with a wide range of diseases and multiple forms of cancer including breast, colon, and prostate cancers. Relatively recent work has demonstrated vitamin D to be critical in immune function and therefore important in inflammatory diseases such as inflammatory bowel disease(IBD). Because vitamin D deficiency or insufficiency is increasingly prevalent around the world, with an estimated 30%-50% of children and adults at risk for vitamin D deficiency worldwide, it could have a significant impact on IBD. Epidemiologic studies suggest that low serum vitamin D levels are a risk factor for IBD and colon cancer, and vitamin D supplementation is associated with decreased colitis disease activity and/or alleviated symptoms. Patients diagnosed with IBD have a higher incidence of colorectal cancer than the general population, which supports the notion that inflammation plays a key role in cancer development and underscores the importance of understanding how vitamin D influences inflammation and its cancer-promoting effects. In addition to human epidemiological data, studies utilizing mouse models of colitis have shown that vitamin D is beneficial in preventing or ameliorating inflammation and clinical disease. The precise role of vitamin D on colitis is unknown; however, vitamin D regulates immune cell trafficking and differentiation, gut barrier function and antimicrobial peptide synthesis, all of which may be protective from IBD and colon cancer. Here we focus on effects of vitamin D on inflammation and inflammation-associated colon cancer and discuss the potential use of vitamin D for protection and treatment of IBD and colon cancer.
基金Supported by Funds received from the National Cancer Institute (CA127481,CA129776)Geyer Foundation,New York,Louisiana Cancer Research Consortium and Tulane Cancer Center
文摘AIM: To develop a hepatocellular carcinoma (HCC) xenograft model for studying hepatitis C virus (HCV) replication in a mice, and antiviral treatment.METHODS: We developed a stable S3-green fluorescence protein (GFP) cell line that replicated the GFP-tagged HCV sub-genomic RNA derived from a highly efficient JFH1 virus. S3-GFP replicon cell line was injected subcutaneously into γ-irradiated SCID mice. We showed that the S3-GFP replicon cell line formed human HCC xenografts in SCID mice. Cells were isolated from subcutaneous tumors and then serially passaged multiple times in SCID mice by culturing in growth medium supplemented with G-418. The mouse-adapted S3-GFP replicon cells were implanted subcutaneously and also into the liver of SCID mice via intrasplenic infusion to study the replication of HCV in the HCC xenografts. The tumor model was validated for antiviral testing after intraperitoneal injection of interferon-α (IFN-α). RESULTS: A highly tumorigenic S3-GFP replicon cell line was developed that formed subcutaneous tumors within 2 wk and diffuse liver metastasis within 4 wk in SCID mice. Replication of HCV in the subcutaneous and liver tumors was confirmed by cell colony assay, detection of the viral RNA by ribonuclease protection assay and real-time quantitative reverse transcription polymerase chain reaction. High-level replication of HCV sub-genomic RNA in the tumor could be visualized by GFP expression using fluorescence microscopy. IFN-α cleared HCV RNA replication in the subcutaneous tumors within 2 wk and 4 wk in the liver tumor model. CONCLUSION: A non-infectious mouse model allows us to study replication of HCV in subcutaneous and metastatic liver tumors. Clearance of HCV by IFN-α supports use of this model to test other anti-HCV drugs.
基金supported by grants from Alberta Innovates/Alberta Prion Research Institute(APRI grants 201600010 and 201900008)(to HMS)ST had received a University of Calgary Eyes High,Killam and Alberta Innovates Health Solution(AIHS)doctoral fellowship.
文摘Prion diseases are infectious protein misfolding disorders of the central nervous system that result from misfolding of the cellular prion protein(PrPC)into the pathologic isoform PrPSc.Pathologic hallmarks of prion disease are depositions of pathological prion protein PrPSc,neuronal loss,spongiform degeneration and astrogliosis in the brain.Prion diseases affect human and animals,there is no effective therapy,and they invariably remain fatal.For a long time,neuronal loss was considered the sole reason for neurodegeneration in prion pathogenesis,and the contribution of non-neuronal cells like microglia and astrocytes was considered less important.Recent evidence suggests that neurodegeneration during prion pathogenesis is a consequence of a complex interplay between neuronal and non-neuronal cells in the brain,but the exact role of these non-neuronal cells during prion pathology is still elusive.Astrocytes are non-neuronal cells that regulate brain homeostasis under physiological conditions.However,astrocytes can deposit PrPSc aggregates and propagate prions in prion-infected brains.Additionally,sub-populations of reactive astrocytes that include neurotrophic and neurotoxic species have been identified,differentially expressed in the brain during prion infection.Revealing the exact role of astrocytes in prion disease is hampered by the lack of in vitro models of prion-infected astrocytes.Recently,we established a murine astrocyte cell line persistently infected with mouse-adapted prions,and showed how such astrocytes differentially process various prion strains.Considering the complexity of the role of astrocytes in prion pathogenesis,we need more in vitro and in vivo models for exploring the contribution of sub-populations of reactive astrocytes,their differential regulation of signaling cascades,and the interaction with neurons and microglia during prion pathogenesis.This will help to establish novel in vivo models and define new therapeutic targets against prion diseases.In this review,we will discuss the complex role of astrocytes in prion disease,the existing experimental resources,the challenges to analyze the contribution of astrocytes in prion disease pathogenesis,and future strategies to improve the understanding of their role in prion disease.
文摘Background:Tiletamine/zolazepam is a dissociative anesthetic combination commonly used in small animals but information is limited in rats.The alpha-2 agonist,dexmedetomidine,has gained popularity in laboratory animal anesthesia.Tramadol is a weak opioid mu agonist.The aim of this study was to assess whether the tiletamine/zolazepam/dexmedetomidine(ZD)combination effectively provides a surgical anesthesia plane comparable to tiletamine/zolazepam/dexmedetomidine with tramadol(ZDT)in a minor procedure in rats.Methods:Rats were induced with ZD or ZDT.After the loss of paw withdrawal,a small incision was made on the rats’left thighs as a surgical stimulus.Rats were maintained under a surgical anesthesia plane by assessing the loss of the paw withdrawal reflex for 45 minutes,then atipamezole was administered.Monitored anesthesia parameters included:(a)physiological parameters-pulse rate(PR),respiratory rate(RR),tissue oxygen saturation(%SpO 2),and body temperature;(b)duration parameters-induction time,onset and duration of surgical anesthesia plane,onset of recovery,and recovery time.Results:PR was significantly lower at 10 minutes in ZD and 5 minutes in ZDT groups.No difference was observed for RR,%SpO 2,and body temperature.Likewise,there were no differences for duration parameters:induction time was less than 3 minutes;onset and duration of surgical anesthesia plane were approximately 5 and 45 minutes,respectively;onset of recovery(time to move)was 51 minutes;and recovery time was 52 minutes,respectively.Conclusion:These data suggest the ZD combination provides a surgical anesthesia plane comparable to ZDT in a rat incisional pain model.
基金the National Institutes of Health(NIH R01 HL089315-01 and NIH R01 HL152155,YJW)the Thoracic Surgery Foundation Resident Research Fellowship(YZ)the National Science Foundation Graduate Research Fellowship Program(AMI).
文摘Resource-scarce regions with serious COVID-19 outbreaks do not have enough ventilators to support critically ill patients,and these shortages are especially devastating in developing countries.To help alleviate this strain,we have designed and tested the accessible low-barrier in vivo-validated economical ventilator(ALIVE Vent),a COVID-19-inspired,cost-effective,open-source,in vivo-validated solution made from commercially available components.The ALIVE Vent operates using compressed oxygen and air to drive inspiration,while two solenoid valves ensure one-way flow and precise cycle timing.The device was functionally tested and profiled using a variable resistance and compliance artificial lung and validated in anesthetized large animals.Our functional test results revealed its effective operation under a wide variety of ventilation conditions defined by the American Association of Respiratory Care guidelines for ventilator stockpiling.The large animal test showed that our ventilator performed similarly if not better than a standard ventilator in maintaining optimal ventilation status.The FiO2,respiratory rate,inspiratory to expiratory time ratio,positive-end expiratory pressure,and peak inspiratory pressure were successfully maintained within normal,clinically validated ranges,and the animals were recovered without any complications.In regions with limited access to ventilators,the ALIVE Vent can help alleviate shortages,and we have ensured that all used materials are publicly available.While this pandemic has elucidated enormous global inequalities in healthcare,innovative,cost-effective solutions aimed at reducing socio-economic barriers,such as the ALIVE Vent,can help enable access to prompt healthcare and life saving technology on a global scale and beyond COVID-19.
文摘Background:Extended-release buprenorphine(XR)is indicated for pain management in rodents,but little is known about its use in mice.This study aimed to investigate whether high dose XR effectively attenuates post-operative hypersensitivity better than low dose XR in a mouse model of incisional pain.Methods:Mice(n 44)were randomly assigned to 1 of 4 treatment groups:(a)saline(1 ml/kg SC,once);(b)sustained release buprenorphine(Bup-SR,1 mg/kg SC,once);(c)low dose extended-release buprenorphine(XR-lo,3.25 mg/kg SC,once);(d)high dose extended-release buprenorphine(XR-hi,6.5 mg/kg SC,once).On days1,0(4 hours),1,2,and 3,mechanical and thermal hypersensitivities were evaluated,and plasma buprenorphine concentrations were measured.Results:Mechanical(days 0-2)and thermal(days 0-1)hypersensitivities were ob-served in the saline group.Bup-SR,XR-lo,and XR-hi attenuated mechanical hyper-sensitivity on days 0,1,and 2.None of the treatment groups,except XR-Lo on day 0,attenuated thermal hypersensitivity on days 0 or 1.Plasma buprenorphine concen-tration peaked at 4 hours(day 0)in all treatment groups and remained greater than 1 ng/mL on days 0-2.No abnormal clinical observations or gross pathologic findings were seen in any groups.Conclusion:The results indicate XR-hi did not effectively attenuate post-operative hypersensitivity better than XR-lo.Thus both 3.25 and 6.5 mg/kg XR are recom-mended for attenuating post-operative hypersensitivity for at least up to 48 hours in mice.
文摘Background and Aim: The porcine heart bears the best resemblance to the human heart and remains the preferred preclinical model for anatomical, physiological, and medical device studies. In an effort to study phenomena related strictly to ischemia reperfusion and donor preservation protocols, it is essential to avoid the immune responses related to allotransplantation. Orthotopic auto-transplantation is a unique strategy to the field of cardiac transplantation for ex vivo experimentation. Nevertheless, auto-transplantation carries its own technical challenges related to insufficient length of the great vessels that are to be transected and re-anastomosed. Methods: A novel method for orthotopic cardiac auto-transplantation in the porcine model was developed and was described herein. Porcine models were used for ex vivo experimentation of a novel device to study ischemia reperfusion injury. Results: A total of five porcine models were used for ex vivo experimentation of a novel device to mitigate ischemia reperfusion injury and determine effects of donor preservation. Modifications to routine cardiac transplantation protocols to allow for successful auto-transplantation are described. Conclusion: Orthotopic cardiac auto-transplantation in the porcine model is a plausible and technically feasible method for reliable study of ischemia reperfusion injury and donor preservation protocols. Here, we describe methods for both direct orthotopic porcine cardiac auto-transplantations as well as a simplified protocol that can be substituted for full surgical auto-transplantation for the studies of preservation of donor hearts.
基金supported by the National Natural Science Foundation of China (51703018, 22375027)the Natural Science Foundation of Jiangsu Province (BK20221265, BK20211100)+1 种基金the Fundamental Research Funds for the Central Universities (DUT21YG133, DUT22YG224)the Research Funds from Liaoning Cancer Hospital(2024ZLKF-35)。
文摘Carbon monoxide(CO) is an endogenous therapeutic gas with an anti-tumor effect. The precise delivery and controlled release of CO in tumor tissues play crucial roles in anti-cancer treatment. However, efficient in situ generation of CO from metal-free COreleasing molecules(CORMs) remains a formidable challenge. Herein, we develop ultrasound(US)-driven self-decomposition porphyrin as organic and metal-free US-CORMs, which can spatiotemporally control the CO release(347 mmol CO/mol porphyrin) efficiently under physiologically harmless US conditions(1.0 MHz, 1.5 W/cm^(2), 50% duty cycle, 50 min). Moreover,porphyrin as a sonosensitizer can also generate reactive oxygen species(ROS) under US treatment to achieve sonodynamic therapy(SDT). Advanced functions of such porphyrin-based CORMs in CO gas-sonodynamic synergistic treatment have been demonstrated by evaluating the in vitro and in vivo anti-tumor effects.
基金K.-P.K.acknowledges support from the Korean Fund for Regenerative Medicine(KFRM)grant funded by the Korea government(the Ministry of Science and ICT,the Ministry of Health&Welfare)(22A0203L1)J.K.acknowledges support from the German Research Council(DFG,Project 496710327)Federal Ministry for Education and Research(BMBF)with the grant number 03ZU1202FA.
文摘In a recent publication in Cell,Li et al.reported that inhibiting the spliceosome in human pluripotent stem cells(hPSCs)can induce a stable totipotent state.1 This study signifies the importance of post-transcriptional regulation in controlling developmental potential and provides a powerful model system for studying the process of human pre-implantation development in vitro.
基金This study was supported by grants from Fujian Province Science and Technology Program （No. 2015Y0026） and Science and Technology Programs of Natural Science Foundation （No. 2016J01585）.
文摘Background： Contribution of model for end-stage liver disease incorporating with serum sodium （MELD-Na） score in predicting acute kidney injury （AKI） after orthotopic liver transplantation （OLT） is yet to be identified. This study assessed the prognostic value ot MELD-Na score for the development of AKl following OLT. Methods： Preoperative and surgery-related variables of 321 adult end-stage liver disease patients who underwent OLT in Fuzhou General Hospital were collected. PostoperativeAKI was defined and staged in accordance with the clinical practice guidelines developed by Kidney Disease： Improving Global Outcomes. Univariate and multivariate analysis was performed to determine the risk factors fnr AKI following OLT. The discriminating power of MELD/MELD-Na score on AKI outcome was evaluated by receiver operating characteristic （ROC） curve. Spearman＇s correlation analysis was used for identifying the correlated relationship between MELD/MELD-Na score and the severity levels of AKI. Results： The prevalence of AKI following OLT was in 206 out of 321 patients （64.2%）. Three risk lhctors for AKI post-OLT were presented, preoperative calculated MELD score （odds ratio [OR] = 1.048, P = 0.021）, intraoperative volume of red cell suspension transfusion （OR = 1.001, P 0.002）, and preoperative liver cirrhosis （OR = 2.015, P = 0.012）. Two areas under ROC curve （AUCs） of MELD/MELD-Na score predicting AKI were 0.688 and 0.672, respectively; the difference between two AUCs was not significant （Z= 1.952, P = 0.051）. The Spearman＇s correlation coefficients between MELD/MELD-Na score and the severity levels of AKI were 0.406 and 0.385 （P 0.001, 0.001）, respectively. Conclusions： We demonstrated that preoperative MELD score, intraoperative volume of red cell suspension transfusion and preoperative liver cirrhosis were risk factors for AKI following OLT. Furthermore, we preliminarily validated that MELD score seemed to have a stronger power discriminating AKI post-OLT than that of novel MELD-Na score.
基金supported by the National Key R&D Program of China(2019YFA0905203)National Natural Science Foundation of China(51703095)+4 种基金Natural Science Foundation of Jiangsu Province(BK20171010)the State Key Laboratory of Materials-Oriented Chemical Engineering(ZK201905)the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture(XTB1804)Jiangsu Agricultural Science and Technology Innovation Fund(CX(19)3115)the China Postdoctoral Science Foundation(2019M661814).
文摘Uncontrolled bleeding and infection can cause significant increases in mortalities.Hydrogel sealants have attracted extensive attention for their ability to control bleeding.However,because interfacial water is a formidable barrier to strong surface bonding,a challenge remains in finding a product that offers robust tissue adhesion combined with anti-infection properties.Inspired by the strong adhesive mechanism of biofilm and mussels,we report a novel dual bionic adhesive hydrogel(DBAH)based on chitosan grafted with methacrylate(CS-MA),dopamine(DA),and N-hydroxymethyl acrylamide(NMA)via a facile radical polymerization process.CS-MA and DA were simultaneously included in the adhesive polymer for imitating the two key adhesive components:polysaccharide intercellular adhesin(PIA)of staphylococci biofilm and 3,4-dihydroxy-L-phenylalanine(Dopa)of mussel foot protein,respectively.DBAH presented strong adhesion at 34 kPa even upon three cycles of full immersion in water and was able to withstand up to 168 mm Hg blood pressure,which is significantly higher than the 60–160 mm Hg measured in most clinical settings.Most importantly,these hydrogels presented outstanding hemostatic capability under wet and dynamic in vivo movements while displaying excellent antibacterial properties and biocompatibility.Therefore,DBAH represents a promising class of biomaterials for high-efficiency hemostasis and wound healing.
基金was supported by grants from National Institutes of Health(R01DK089098 and P01DK57751)American Diabetes Association(1-19-IBS-119)+1 种基金X.Y.and a Glenn/AFAR Scholarship for Research in the Biology of Aging to M.-D.LYale School of Medicine and by the Office of The Director,National Institutes of Health(S10OD02365101A1,S10OD019967,and S10OD018034).
文摘O-linked N-acetyl-glucosamine glycosylation(O-GlcNAcylation)of intracellular proteins is a dynamic process broadly implicated in age-related disease,yet it remains uncharacterized whether and how O-GlcNAcylation contributes to the natural aging process.O-GlcNAc transferase(OGT)and the opposing enzyme O-GlcNAcase(OGA)control this nutrient-sensing protein modification in cells.Here,we show that global O-GlcNAc levels are increased in multiple tissues of aged mice.In aged liver,carbamoyl phosphate synthetase 1(CPS1)is among the most heavilyO-GlcNAcylated proteins.CPS1O-GlcNAcylation is reversed by calorie restriction and is sensitive to genetic and pharmacological manipulations of theO-GlcNAc pathway.High glucose stimulates CPS1O-GlcNAcylation and inhibits CPS1 activity.Liver-specific deletion of OGT potentiates CPS1 activity and renders CPS1 irresponsive to further stimulation by a prolonged fasting.Our results identify CPS1 O-GlcNAcylation as a key nutrient-sensing regulatory step in the urea cycle during aging and dietary restriction,implying a role for mitochondrial O-GlcNAcylation in nutritional regulation of longevity.
基金supported by The Beljanski Foundation(to JY)and Military Laboratory Animal Fund(Grant No.SYDW[2017]15to TF)。
文摘Objective:Herbal medicine is an important therapeutic option for benign prostatic hyperplasia(BPH),a common disease in older men that can seriously affect their quality of life.Currently,it is crucial to develop agents with strong efficacy and few side effects.Herein we investigated the effects of the extract of Rauwolfia vomitoria,a shrub grown in West Africa,on BPH.Methods:Rats with testosterone-induced BPH were treated with R.vomitoria.Prostates were histologically analyzed by Hematoxylin and eosin staining.Proliferation index and the expression levels of androgen receptor and its associated proteins were quantified through immunohistochemistry and immunoblotting.Androgen receptor target genes were examined by quantitative real-time polymerase chain reaction.The sperm count and body weight of rats were also measured.Results:The oral administration of R.vomitoria extract significantly reduced the prostate weight and prostate weight index in BPH rats,supported by the decreased thickness of the prostate epithelial layer and increased lumen size.Similar effects were observed in the BPH rats treated with the reference drug,finasteride.R.vomitoria extract significantly reduced the testosterone-induced proliferation markers,including proliferating cell nuclear antigen and cyclin D1,in the prostate glands of BPH rats;it also reduced levels of androgen receptor,its associated protein steroid 5 a-reductase 1 and its downstream target genes(FK506-binding protein 5 and matrix metalloproteinase 2).Notably,compared with the finasteride group,R.vomitoria extract did not significantly reduce sperm count.Conclusion:R.vomitoria suppresses testosterone-induced BPH development.Due to its milder side effects,R.vomitoria could be a promising therapeutic agent for BPH.
基金a Cancer Research Society award(Scholarships for the New Generation of Scientists).NJB was supported by Multiple Myeloma Research Foundation.ATS was supported by the National Institutes of Health(NIHGrant No.K08CA230188)+2 种基金the Parker Institute for Cancer Immunotherapy,a Cancer Research Institute Technology Impact Award,a Career Award for Medical Scientists from the Burroughs Wellcome Fund,and the Human Vaccines Project Michelson Prize.HJ and WZ were supported by the National Human Genome Research Institute of NIH(Grant Nos.R01HG009518 and R01HG010889)JAS was supported by an Alberta Children’s Hospital Research Institute Postdoctoral Fellowship.AJ was supported by an Alberta Innovates Summer Studentship.JB was supported by Canadian Institutes of Health Research(PJT-401394)Calgary Firefighters Burn Treatment Society.
文摘How distinct transcriptional programs are enacted to generate cellular heterogeneity and plasticity,and enable complex fate decisions are important open questions.One key regulator is the cell’s epigenome state that drives distinct transcriptional programs by regulating chromatin accessibility.Genome-wide chromatin accessibility measurements can impart insights into regulatory sequences(in)accessible to DNA-binding proteins at a single-cell resolution.This review outlines molecular methods and bioinformatic tools for capturing cell-to-cell chromatin variation using single-cell assay for transposase-accessible chromatin using sequencing(scATAC-seq)in a scalable fashion.It also covers joint profiling of chromatin with transcriptome/proteome measurements,computational strategies to integrate multi-omic measurements,and predictive bioinformatic tools to infer chromatin accessibility from single-cell transcriptomic datasets.Methodological refinements that increase power for cell discovery through robust chromatin coverage and integrate measurements from multiple modalities will further expand our understanding of gene regulation during homeostasis and disease.
基金This study was supported by a National Science,Technology,and Innovation plan(NSTIP)project grant(15-MED4186-20)awarded by the King Abdulaziz City for Science and Technology(KACST)to JK,AMA,and FAL.
文摘Sperm-specific phospholipase C zeta(PLCζ)initiates intracellular calcium(Ca2+)transients which drive a series of concurrent events collectively termed oocyte activation.Numerous investigations have linked abrogation and absence/reduction of PLCζwith forms of male infertility in humans where oocyte activation fails.However,very few studies have examined potential relationships between PLCζand advancing male age,both of which are increasingly considered to be major effectors of male fertility.Initial efforts in humans may be hindered by inherent PLCζvariability within the human population,alongside a lack of sufficient controllable repeats.Herein,utilizing immunoblotting,immunofluorescence,and quantitative reverse transcription PCR(qRT-PCR)we examined for the first time PLCζprotein levels and localization patterns in sperm,and PLCζmRNA levels within testes,from mice at 8 weeks,12 weeks,24 weeks,and 36 weeks of age,from two separate strains of mice,C57BL/6(B6;inbred)and CD1(outbred).Collectively,advancing male age generally diminished levels and variability of PLCζprotein and mRNA in sperm and testes,respectively,when both strains were examined.Furthermore,advancing male age altered the predominant pattern of PLCζlocalization in mouse sperm,with younger mice exhibiting predominantly post-acrosomal,and older mice exhibiting both post-acrosomal and acrosomal populations of PLCζ.However,the specific pattern of such decline in levels of protein and mRNA was strain-specific.Collectively,our results demonstrate a negative relationship between advancing male age and PLCζlevels and localization patterns,indicating that aging male mice from different strains may serve as useful models to investigate PLCζin cases of male infertility and subfertility in humans.
基金This work was supported by a COVID-19 project grant(#951)awarded by the Saudi Arabian Ministry of Health awarded to JK,KAK,and AY.
文摘A recent study published in Science by Shrock et al.1 examined how responses to COVID-19 severity differed amongst patients with different prior viral exposure history,finding notable correlations between both.Such serological profiling provides a window into differential viral responses amongst patients with diverse outcomes,potentially mediating improved therapeutics and vaccines for SARS-CoV-2.
