Echinacoside attenuates glandular fibrosis in benign prostatic hyperplasia via inhibiting MKK6/MK2 signaling pathway

Background:Lower urinary tract symptoms commonly occur in the elderly population and seriously constrain the quality of life.Glandular fibrosis is an important pathobiological process in benign prostatic hyperplasia and is also a main inducing factor for benign prostatic hyperplasia-associated lower urinary tract symptoms.Cistanches species is an important herbal medicine resource and is traditionally used in ameliorating renal and prostatic defects.Methods:This study was to investigate the potential protective function of echinacoside(a bioactive compound from Cistanches)against prostatic fibrosis in mice and human benign prostatic hyperplasia epithelial-1 cell models.Results:It was found that echinacoside attenuated testosterone-induced prostatic hyperplasia and collagen deposition in mice,relieved prostate local inflammation and oxidative damage,and ameliorated prostatic epithelial-mesenchymal transition.Additionally,echinacoside inhibited the activation of the MKK6/MK2 signaling pathway both in vivo and in vitro.Conclusion:This study added new evidence for the anti-fibrotic function of echinacoside on the prostate and provided new insights for understanding its possible pharmacological mechanisms.

Therapeutic effects of Lingguizhugan decoction in a rat model of high-fat diet-induced insulin resistance

BACKGROUND Lingguizhugan(LGZG)decoction is a widely used classic Chinese medicine formula that was recently shown to improve high-fat diet(HFD)-induced insulin resistance(IR)in animal studies.AIM To assess the therapeutic effect of LGZG decoction on HFD-induced IR and explore the potential underlying mechanism.METHODS To establish an IR rat model,a 12-wk HFD was administered,followed by a 4-wk treatment with LGZG.The determination of IR status was achieved through the use of biochemical tests and oral glucose tolerance tests.Using a targeted metabolomics platform to analyze changes in serum metabolites,quantitative real-time PCR(qRT-PCR)was used to assess the gene expression of the ribosomal protein S6 kinase beta 1(S6K1).RESULTS In IR rats,LGZG decreased body weight and indices of hepatic steatosis.It effectively controlled blood glucose and food intake while protecting islet cells.Metabolite analysis revealed significant differences between the HFD and HFDLGZG groups.LGZG intervention reduced branched-chain amino acid levels.Levels of IR-related metabolites such as tryptophan,alanine,taurine,and asparagine decreased significantly.IR may be linked to amino acids due to the contemporaneous increase in S6K1 expression,as shown by qRT-PCR.CONCLUSIONS Our study strongly suggests that LGZG decoction reduces HFD-induced IR.LGZG may activate S6K1 via metabolic pathways.These findings lay the groundwork for the potential of LGZG as an IR treatment.

Repetitive traumatic brain injury–induced complement C1–related inflammation impairs long-term hippocampal neurogenesis

Repetitive traumatic brain injury impacts adult neurogenesis in the hippocampal dentate gyrus,leading to long-term cognitive impairment.However,the mechanism underlying this neurogenesis impairment remains unknown.In this study,we established a male mouse model of repetitive traumatic brain injury and performed long-term evaluation of neurogenesis of the hippocampal dentate gyrus after repetitive traumatic brain injury.Our results showed that repetitive traumatic brain injury inhibited neural stem cell proliferation and development,delayed neuronal maturation,and reduced the complexity of neuronal dendrites and spines.Mice with repetitive traumatic brain injuryalso showed deficits in spatial memory retrieval.Moreover,following repetitive traumatic brain injury,neuroinflammation was enhanced in the neurogenesis microenvironment where C1q levels were increased,C1q binding protein levels were decreased,and canonical Wnt/β-catenin signaling was downregulated.An inhibitor of C1 reversed the long-term impairment of neurogenesis induced by repetitive traumatic brain injury and improved neurological function.These findings suggest that repetitive traumatic brain injury–induced C1-related inflammation impairs long-term neurogenesis in the dentate gyrus and contributes to spatial memory retrieval dysfunction.

OpenNAU:An open-source platform for normalizing,analyzing,and visualizing cancer untargeted metabolomics data

Objective:As an important part of metabolomics analysis,untargeted metabolomics has become a powerful tool in the study of tumor mechanisms and the discovery of metabolic markers with high-throughput spectrometric data which also poses great challenges to data analysis,from the extraction of raw data to the identification of differential metabolites.To date,a large number of analytical tools and processes have been developed and constructed to serve untargeted metabolomics research.The different selection of analytical tools and parameter settings lead to varied results of untargeted metabolomics data.Our goal is to establish an easily operated platform and obtain a repeatable analysis result.Methods:We used the R language basic environment to construct the preprocessing system of the original data and the LAMP(Linux+Apache+MySQL+PHP)architecture to build a cloud mass spectrum data analysis system.Results:An open-source analysis software for untargeted metabolomics data(openNAU)was constructed.It includes the extraction of raw mass data and quality control for the identification of differential metabolic ion peaks.A reference metabolomics database based on public databases was also constructed.Conclusions:A complete analysis system platform for untargeted metabolomics was established.This platform provides a complete template interface for the addition and updating of the analysis process,so we can finish complex analyses of untargeted metabolomics with simple human-computer interactions.The source code can be downloaded from https://github.com/zjuRong/openNAU.

The role of periodontal disease in atherosclerotic cardiovascular disease

Atherosclerotic cardiovascular disease(ASCVD)includes a group of disorders of the heart and blood vessels and accounts for major morbidity and premature death worldwide.Periodontitis is a chronic inflammatory disease with the gradual destruction of supporting tissues around the teeth,including gingiva,periodontal ligament,alveolar bone,and cementum.Periodontitis has been found to potentially increase the risk of ASCVD.Generally,oral microorganisms and inflammation are the major factors for periodontitis to the incidence of ASCVD.Recently,evidence has shown that the loss of masticatory function is another important factor of periodontitis to the incidence of ASCVD.In this review,we illustrate the recent finding of the relationship between periodontitis and ASCVD,from a microscale perspective-oral microorganisms,inflammation,and tooth loss.With the high prevalence of periodontitis,it is important to add oral therapy as a regular ASCVD prevention strategy.Regular dental visits could be a helpful strategy for ASCVD patients or general medical practitioners.

Evaluation of Ultrasound-guided Genitofemoral Nerve Block Combined with Ilioinguinal/iliohypogastric Nerve Block during Inguinal Hernia Repair in the Elderly

To evaluate the anesthetic effect of ultrasound-guided(USG)ilioinguinal/iliohypogastric nerve(Ⅱ/IHN)block combined with genital branch of genitofemoral nerve(GFN)block in the elderly undergoing inguinal hernia repair,54 old patients(aged 60-96years,ASAⅠ-Ⅲ)with indirect hernia were enrolled and scheduled for unilateral tensiofree herniorrhaphy.Patients were grouped randomly to receive either USGⅡ/IHN plus GFN block(Group G)or USGⅡ/IHN block alone(GroupⅠ).The intraoperative visual analogue scale(VAS)scores were recorded at skin incision,at spermatic cord/round ligament traction and at sac ligation.The resting and dynamic VAS scores were recorded postoperatively.The requirements of extra sedatives and analgesics for intra-and postoperative analgesia were assessed.Occurrence of complications of the block,postoperative nausea and vomiting and femoral nerve palsy was also reported.Both groups showed similar sensory block.When stretching spermatic cord/round ligament,the patients in group G had significantly lower VAS scores than in group.And group G used much fewer adjuvant sedatives and analgesics to achieve adequate anaesthesia.In addition,group G was presented with better intraoperative anaesthesia and lower postoperative dynamic VAS scores at all time points tested.No significant difference was found in the postoperative requirement of rescue medication.Both groups showed no complications related to the block and group G reported no femoral nerve palsy.The addition of GFN block toⅡ/IHN block improves the quality of perioperative anesthesia and analgesia in the elderly and reduces the consumption of extra sedatives and analgesics during the surgery.

Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B

Objective: To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be used to find the relevant genes and interesting proteins associated with the development of hepatitis B. Methods: The total RNA from liver tissue with chronic hepa- titis B was extracted and the mRNA was purified using TRIZOL method. Switching mechanism at 5′ end of the RNA transcript (SMART) technique and CDS III/3′ primer were used for first-strand cDNA synthesis. Long distance polymerase chain reaction (LD PCR) was then used to synthesize the double-strand cDNA that was then digested by Sfi I and fractionated by CHROMA SPIN-400 column. The longer than 0.4 kb cDNAs were collected and ligated to λTriplEx2 vector. Then λ phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. Fourteen plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. Results: The titers of unamplifed and amplified libraries were 1.94×106 pfu/ml and 1.49×109 pfu/ml respectively. The percentages of recombinants from both libraries were 98.15% in unamplified library and 98.76% in amplified library. The lengths of the inserts were 1.23 kb in average, 1?2 kb in 64.29%, and 0.5?1.0 kb in 35.71%. Conclusion: A high quality cDNA library from human liver tissue with chronic hepatitis B was successfully constructed.

The effect of the Formative evaluation on the medical student＇s career emotion, practicing skills and social adaptation

With the development of the knowledge economy, social professional quality of talent has an increasingly demanding. Medical students is China＇ s medical and health undertakings reserve personnel, it plays an important role for better protection of people＇ s health by strengthening vocational personality training medical students and improving professional quality. Therefore, we must pay attention to the medical student＇ s career awareness, ethics, professionalism, career aspirations and career interests of education on medical students to develop a healthy personality of the medical profession, and lay the foundation for the sustainable development of medical and health services.

The Role of the Examination Management for Medical Students in Humanities Education and Training Professional Quality

It has been proved by years of research that if the function of examination management cannot be fully understood or be used effectively, the humanities education and train of professional quality for medical students will not make a fundamental achievement. Because it is not only an academic training but also the examination management and the humanities cultivation have a close connection. The humanities cultivation which is based on the core of virtue requires a good campus environment. Therefore, we should adjust to the medical students＇ humanities quality training requirements, making the targeted reform and innovation of examination management.

A Cullin 5-based complex serves as an essential modulator of ORF9b stability in SARS-CoV-2 replication

The ORF9b protein,derived from the nucleocapsid’s open-reading frame in both SARS-CoV and SARS-CoV-2,serves as an accessory protein crucial for viral immune evasion by inhibiting the innate immune response.Despite its significance,the precise regulatory mechanisms underlying its function remain elusive.In the present study,we unveil that the ORF9b protein of SARS-CoV-2,including emerging mutant strains like Delta and Omicron,can undergo ubiquitination at the K67 site and subsequent degradation via the proteasome pathway,despite certain mutations present among these strains.Moreover,our investigation further uncovers the pivotal role of the translocase of the outer mitochondrial membrane 70(TOM70)as a substrate receptor,bridging ORF9b with heat shock protein 90 alpha(HSP90α)and Cullin 5(CUL5)to form a complex.Within this complex,CUL5 triggers the ubiquitination and degradation of ORF9b,acting as a host antiviral factor,while HSP90αfunctions to stabilize it.Notably,treatment with HSP90 inhibitors such as GA or 17-AAG accelerates the degradation of ORF9b,leading to a pronounced inhibition of SARS-CoV-2 replication.Single-cell sequencing data revealed an up-regulation of HSP90αin lung epithelial cells from COVID-19 patients,suggesting a potential mechanism by which SARS-CoV-2 may exploit HSP90αto evade the host immunity.Our study identifies the CUL5-TOM70-HSP90αcomplex as a critical regulator of ORF9b protein stability,shedding light on the intricate host–virus immune response dynamics and offering promising avenues for drug development against SARS-CoV-2 in clinical settings.

Photothermal effective CeO_(2)NPs combined in thermosensitive hydrogels with enhanced antibacterial,antioxidant and vascularization performance to accelerate infected diabetic wound healing

Chronic diabetic wound healing remains a formidable challenge due to susceptibility to bacterial infection,excessive oxidative stress,and poor angiogenesis.To address these issues,a sodium alginate(SA)based photothermal hydrogel dressing with multifunction was fabricated to facilitate wound treatment.Ceria nanoparticles(CeO_(2)NPs)was synthesized,and their antibacterial performance by near-infrared light triggered photothermal effects was first studied and verified in this work.In addition,to release CeO_(2)NPs to achieve antioxidation and pro-vascularization,thermosensitive gelatin(Gel)was utilized to embed the nanoparticles in advance and then composited in SA hydrogel networks.SA network was finally strengthened by acid soaking to form partially crystalline regions to act as natural crosslinkers.Results showed that the Gel/SA/CeO_(2)hydrogel displayed temperature-responsive release of CeO_(2)NPs,significant antibacterial and antioxidative activity,as well as the ability to remove without injury and promote infected diabetic wound healing with low cytotoxicity,according to antibacterial investigations,cell studies,and in vivo animal studies.This research offers not only a successful method for quickening the healing of diabetic wounds but also a fresh approach to the general use of CeO_(2)NPs.

Gene Expressions Underlying Mishandled Calcium Clearance and Elevated Generation of Reactive Oxygen Species in the Coronary Artery Smooth Muscle Cells of Chronic Heart Failure Rats

Background： The calcium clearance and reactive oxygen species （ROS） generations in the coronary artery smooth muscle cells in chronic heart failure （HF） have not been fully investigated. Therefore, we attempted to understand the gene expressions underlying the mishandling of calcium clearance and the accumulations of ROS. Methods： We initially established an animal model of chronic HF by making the left anterior descending coronary artery ligation （CAL） in rats, and then isolated the coronary artery vascular smooth muscle cells from the ischemic and the nonischemic parts of the coronary artery vessels in 12 weeks after CAL operation. The intracellular calcium concentration and ROS level were measured using flow cytometry, and the gene expressions of sarco/endoplasmic reticulum Ca2＋-ATPase （SERCA2a）, encoding sarcoplasmic reticulum Ca2＋-ATPase 2a, encoding sodium-calcium exchanger （NCX）, andp47phox encoding a subunit of the nicotinamide adenine dinucleotide phosphate （NADPH） oxidase were examined using real-time quantitative reverse transcription polymerase chain reaction and Western blotting, respectively. Results： We found that the calcium accumulation and ROS generation in the coronary artery smooth muscle cells isolated from either the ischemic or the nonischemic part of the CAL coronary artery vessel were significantly increased irrespective of blood supply （all P 〈 0.01 ）. Moreover, these were accompanied by the increased expressions of NCX and p47phox, the decreased expression of S ERCA2a, and the increased amount of phosphorylated forms of p47phox in NADPH oxidase （all P 〈 0.05）. Conclusions： Our results demonstrated that the disordered calcium clearance and the increased ROS generation occurred in the coronary artery smooth muscle cells in rats with chronic HF produced by ligation of the left anterior descending coronary artery （CAL）, and which was found to be disassociated from blood supply, and the increased generation of ROS in the ceils was found to make concomitancy to the increased activity of NADPH oxidase in cytoplasm.

Blocking CD38-driven fratricide among T cells enables effective antitumor activity by CD38-specific chimeric antigen receptor T cells

Chimeric antigen receptor T-cell(CAR T) therapy is a kind of effective cancer immunotherapy. However,designing CARs remains a challenge because many targetable antigens are shared by T cells and tumor cells. This shared expression of antigens can cause CAR T cell fratricide. CD38-targeting approaches(e.g.,daratumumab) have been used in clinical therapy and have shown promising results. CD38 is a kind of surface glycoprotein present in a variety of cells, such as T lymphocytes and tumor cells. It was previously reported that CD38-based CAR T cells may undergo apoptosis or T cell-mediated killing(fratricide) during cell manufacturing. In this study, a CAR containing a sequence targeting human CD38 was designed to be functional. To avoid fratricide driven by CD38 and ensure the production of CAR T cells, two distinct strategies based on antibodies(clone MM12 T or clone MM27) or proteins(H02 H or H08 H) were used to block CD38 or the CAR single-chain variable fragment(scFv) domain, respectively, on the T cell surface.The results indicated that the antibodies or proteins, especially the antibody MM27, could affect CAR T cells by inhibiting fratricide while promoting expansion and enrichment. Anti-CD38 CAR T cells exhibited robust and specific cytotoxicity to CD38+ cell lines and tumor cells. Furthermore, the levels of the proinflammatory factors TNF-a, IFN-g and IL-2 were significantly upregulated in the supernatants of A549CD38+ cells. Finally, significant control of disease progression was demonstrated in xenograft mouse models. In conclusion, these findings will help to further enhance the expansion, persistence and function of anti-CD38 CAR T cells in subsequent clinical trials.

Treatment of CD20-directed Chimeric Antigen Receptor-modified T cells in patients with relapsed or refractory B-cell non-Hodgkin lymphoma:an early phase IIa trial report

Patients with relapsed or refractory non-Hodgkin lymphoma have a dismal prognosis.Chimeric Antigen Receptor(CAR)-modified T cells(CART cells)that targeted CD20 were effective in a phase I clinical trial for patients with advanced B-cell lymphomas.We performed a phase IIa trial to further assess the safety and efficacy of administering autologous anti-CD20 CART(CART-20)cells to patients with refractory or relapsed CD20^(+)B-cell lymphoma.Eleven patients were enrolled,and seven patients underwent cytoreductive chemotherapy to debulk the tumors and deplete the lymphocytes before receiving T-cell infusions.The overall objective response rate was 9 of 11(81.8%),with 6 complete remissions(CRs)and 3 partial remissions;no severe toxicity was observed.The median progression-free survival lasted for 46 months,and 1 patient had a 27-month continuous CR.A significant inverse correlation between the levels of the CAR gene and disease recurrence or progression was observed.Clinically,the lesions in special sites,specifically the spleen and testicle,were refractory to CART-20 treatment.Collectively,these results together with our data from phase I strongly demonstrated the feasibility and efficacy of CART-20 treatment in lymphomas and suggest large-scale patient recruitment in a future study.This study was registered at www.clinicaltrials.org as NCT01735604.

LILRB4 ITIMs mediate the T cell suppression and infiltration of acute myeloid leukemia cells

We recently demonstrated that leukocyte Ig-like receptor 4(LILRB4)expressed by monocytic acute myeloid leukemia(AML)cells mediates T-cell inhibition and leukemia cell infiltration via its intracellular domain.The cytoplasmic domain of LILRB4 contains three immunoreceptor tyrosine-based inhibitory motifs(ITIMs);the tyrosines at positions 360,412,and 442 are phosphorylation sites.Here,we analyzed how the ITIMs of LILRB4 in AML cells mediate its function.Our in vitro and in vivo data show that Y412 and Y442,but not Y360,of LILRB4 are required for T-cell inhibition,and all three ITIMs are needed for leukemia cell infiltration.We constructed chimeric proteins containing the extracellular domain of LILRB4 and the intracellular domain of LILRB1 and vice versa.The intracellular domain of LILRB4,but not that of LILRB1,mediates T-cell suppression and AML cell migration.Our studies thus defined the unique signaling roles of LILRB4 ITIMs in AML cells.

DNA methylation-mediated repression of miR-181a/135a/302c expression promotes the microsatellite-unstable colorectal cancer development and 5-FU resistance via targeting PLAG1

Micro sate Hite instability（MSI） defines a subtype of colorectal cancer（CRC） with typical clinicopathologic characteristics. CRCs with MSI（MSI CRCs） frequently acquire accelerated carcinogenesis and 5-FU resistance, and the exact underlying mechanism remains incompletely understood. Our previous study has identified the microRNA（miRNA） expression profile in MSI CRCs. In this study, three miRNAs（miR-181 a, miR-135 a and miR-302 c） were validated by qRT-PCR to be dramatically decreased in 67 CRC samples. Proliferation and apoptosis assays demonstrated that miR-181 a/135 a/302 c function as tumor suppressors via repressing PLAG1/IGF2 signaling. Moreover, we presented compelling evidence that restoration of miR-181 a/135 a/302 c expression promoted sensitivity of MSI CRC cells to 5-FU treatment. miR-181 a/135 a/302 c exerted their effect on chemoresistance through attenuating PLAG1 expression. Notably, the hypermethylation status of MSI CRC accounts for the decrements of miR-181 a/135 a/302 c. Our results contribute to a better understanding of the mechanism of chemo？resistance in MSI CRCs, and provide a clue for digging the bio markers and therapeutic targets for CRC patients.

An analytical biomarker for treatment of patients with recurrent B-ALL after remission induced by infusion of anti-CD19 chimeric antigen receptor T(CAR-T) cells

Anti-CD19 chimeric antigen receptor-modified T(CAR-T-19) cells have emerged as a powerful targeted immunotherapy for B-cell lineage acute lymphoblastic leukemia with a remarkable clinical response in recent trials. Nonetheless, few data are available on the subsequent clinical monitoring and treatment of the patients, especially those with disease recurrence after CAR-T-19 cell infusion. Here, we analyzed three patients who survived after our phase I clinical trial and who were studied by means of biomarkers reflecting persistence of CAR-T-19 cells in vivo and predictive factors directing further treatment. One patient achieved 9-week sustained complete remission and subsequently received an allogeneic hematopoietic stem cell transplant. Another patient who showed relapse after 20 weeks without detectable leukemia in the cerebrospinal fluid after CAR-T-19 cell treatment was able to achieve a morphological remission under the influence of stand-alone low-dose chemotherapeutic agents. The third patient gradually developed extensive extramedullary involvement in tissues with scarce immune-cell infiltration during a long period of hematopoietic remission after CAR-T-19 cell therapy. Long-term and discontinuous increases in serum cytokines(mainly interleukin 6 and C-reactive protein) were identified in two patients(Nos. 1 and 6) even though only a low copy number of CAR molecules could be detected in their peripheral blood. This finding was suggestive of persistent functional activity of CAR-T-19 cells. Combined analyses of laboratory biomarkers with their clinical manifestations before and after salvage treatment showed that the persistent immunosurveillance mediated by CAR-T-19 cells would inevitably potentiate the leukemia-killing effectiveness of subsequent chemotherapy in patients who showed relapse after CAR-T-19-induced remission.

Long-term safety and efficacy of CART-20 cells in patients with refractory or relapsed B-cell non-Hodgkin lymphoma:5-years follow-up results of the phase I and IIa trials

For refractory or relapsed(r/r)B-cell non-Hodgkin lymphoma(NHL),the response rates to conventional salvage chemotherapy are 27–44%.1 Chimeric antigen receptors(CARs)efficiently redirect T-cell specificity and cytotoxicity to cells expressing the targeted Ag in an HLA-independent manner.2 The early phase clinical trials of CART cells for(r/r)B-cell NHL have demonstrated promising results..

The shape effect of reconstituted high-density lipoprotein nanocarriers on brain delivery and Aβ clearance

Accumulation of extracellular β-amyloid （Aβ） is crucial for the pathogenesis of Alzheimer＇s disease （AD）, and the development of novel therapeutic agents that can both accelerate Aβ clearance and inhibit the subsequent pathological cascades is regarded as a promising strategy for AD management. In our previous study, we have constructed discoidal apolipoprotein E3-reconstituted high-density lipoprotein （ApoE3-rHDL） as an efficient nanoplatform that can penetrate the blood-brain barrier and accelerate Aβ clearance for a combination treatment of AD. To further improve its drug loading capacity, we hypothesized that spherical rHDL might serve as a more powerful nanocarrier if it has the same brain delivery and Aβ clearance abilities as the discoidal rHDL does. To evaluate the potential of spherical rHDL as a promising alternative for the combination therapy for AD, here, we investigated the effect of the shape of rHDL on its brain delivery, Aβ clearance, and anti-AD efficacy. We found that spherical rHDL had stronger Aβ-binding affinity than discoidal rHDL did, more effectively facilitated microglial uptake and degradation of Aβ-42, achieved better brain distribution after intravenous administration, and more powerfully reduced Aβ deposition, decreased microglia activation, attenuated neurological damage, and rescued memory deficits in a mouse model of AD. Among the rHDLs evaluated, monosialotetrahexosyl ganglioside-incorporated spherical rHDL exerted the best effect. The findings of this study for the first time show a shape effect of an rHDL nanocarrier on its biological functions and suggest that a spherical lipoprotein-mimic nanocarrier may serve as a more efficient multifunctional nanoplatforrn for AD therapy.

The macrodomain family:Rethinking an ancient domain from evolutionary perspectives

The reasons why certain domains evolve much slower than others is unclear.The notion that functionally more important genes evolve more slowly than less important genes is one of the few commonly believed principles of molecular evolution.The macrodomain(also known as the X domain) is an ancient,slowly evolving and highly conserved structural domain found in proteins throughout all of the kingdoms and was first discovered nearly two decades ago with the isolation and cloning of macroH2A1.Macrodomains,which are functionally promiscuous,have been studied intensively for the past decade due to their importance in the regulation of cellular responses to DNA damage,chromatin remodeling,transcription and tumorigenesis.Recent structural,phylogenetic and biological analyses,however,suggest the need for some reconsideration of the evolutionary advantage of concentrating such a plethora of diverse functions into the macrodomain and of how macrodomains could perform so many functions.In this article,we focus on macrodomains that are evolving slowly and broadly discuss the potential relationship between the biological evolution and functional diversity of macrodomains.