1 If you visit Lopburi,you might feel like you are on the set of Rise of the Planet of the Apes.While James Franco isnt wandering down the streets,the city is overrun with so many monkeys that you might wonder if they...1 If you visit Lopburi,you might feel like you are on the set of Rise of the Planet of the Apes.While James Franco isnt wandering down the streets,the city is overrun with so many monkeys that you might wonder if theyre taking over.These arent just any monkeys;theyre lively,curious,and absolutely everywhere.展开更多
Complex brain diseases seriously endanger human health,and early diagnostic biomarkers and effective treatments are currently lacking.Due to ethical constraints on human research,establishing monkey models is crucial ...Complex brain diseases seriously endanger human health,and early diagnostic biomarkers and effective treatments are currently lacking.Due to ethical constraints on human research,establishing monkey models is crucial to address these issues.With the rapid development of technology,transgenic monkey models of a range of brain diseases,especially autism spectrum disorder(ASD),have been successfully established.However,to establish practical and effective brain disease models and subsequently apply them to disease mechanism and treatment studies,there is still a lack of a standard tool,i.e.,a system for collecting and analyzing the daily behaviors of brain disease model monkeys.Therefore,with the goal of undertaking a comprehensive and quantitative study of behavioral phenotypes,we established a standard daily behavior collection and analysis system,including behavioral data collection protocols and a monkey daily behavior ethogram(MDBE)for rhesus and cynomolgus monkeys,which are the most commonly used non-human primates in model construction.Then,we used ASD as an application example after referring to the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition,Text Revision(DSM-5-TR),which is widely used in clinical disease diagnosis to obtain ASD core clinical symptoms.We then established a sub-ethogram(ASD monkey core behavior ethogram(MCBE-ASD))specifically for quantitative assessment of the core clinical symptoms of an ASD monkey model based on MDBE.Subsequently,we demonstrated the high reproducibility of the system.展开更多
Background:Non-human primates(NPHs),such as rhesus macaques,cynomol-gus monkeys,and Assamese macaques,play a crucial role in biomedical research.However,baseline cytokine and electrolyte data for these three species,p...Background:Non-human primates(NPHs),such as rhesus macaques,cynomol-gus monkeys,and Assamese macaques,play a crucial role in biomedical research.However,baseline cytokine and electrolyte data for these three species,particularly data stratified by age and sex,are limited.Therefore,the aim of this study was to establish and analyze age-and sex-specific cytokine and electrolyte profiles in these three species.Methods:This study included 40 rhesus macaques(21 males,19 females),33 cyn-omolgus monkeys(17 males,16 females),and 45 Assamese macaques(25 males,20 females)classified by age(1-5 years,6-12 years,>13 years)and sex.The levels of 23 immune function indicators and 5 electrolyte indicators were measured.Results:Among the three monkey species,the levels of sCD40L,IL-18,MCP-1,MIP-1β,TGFa,K^(+),Na^(+),and Cl^(-)exhibited species-,sex-,and age-related differences.Comparison within the same species,sex had no significant impact on cytokine levels in NHPs but did affect electrolyte levels,particularly Cl^(-)and Na^(+)levels,in cynomol-gus monkeys and Assamese macaques.Electrolyte levels in NHPs were not affected by age,whereas the levels of certain cytokines,particularly sCD40L,GM-CSF,and IL-10,varied with age.The remaining 21 cytokines demonstrated no significant age-related changes.Conclusions:Significant variations in cytokine and electrolyte levels exist among dif-ferent monkey species,sexes,and age groups.This research provides valuable re-sources for NHP researchers and sets the stage for further exploring the impacts of sex and age on NHP physiology and immune function.展开更多
While viral infections can disturb the host gut microbiome,the dynamic alterations in microbial composition following infection remain poorly characterized.This study identified SRV-8-infected monkeys and classified t...While viral infections can disturb the host gut microbiome,the dynamic alterations in microbial composition following infection remain poorly characterized.This study identified SRV-8-infected monkeys and classified them into five groups based on infection progression.16S rRNA amplicon sequencing revealed significant alterations in the relative and inferred absolute abundance of bacterial genera UCG-002,Agathobacter,Coprococcus,and Holdemanella during the early stage of SRV-8 infection,coinciding with provirus formation.These microbial shifts were accompanied by functional modifications in bacterial communities at the same stage.In contrast,ITS amplicon sequencing indicated no significant differences in fungal composition between healthy wild-type and SRV-8-infected monkeys.Spearman correlation analyses demonstrated close interactions between intestinal bacteria and fungi following SRV-8 infection.Additionally,SRV-8 seropositive groups exhibited significantly elevated mRNA expression levels of pro-inflammatory(TNF-α,IFN-γ,IL-1β,and IL-6)and anti-inflammatory(IL-10)cytokine genes,highlighting close associations between inflammatory cytokines and immune responses.Overall,these findings provide a comprehensive characterization of bacterial and fungal microbiota dynamics and inflammatory cytokine responses associated with SRV-8 infection,clarifying the pathobiological mechanisms underlying SRV-8 infection from the perspective of the gut microbiome.展开更多
Nonhuman primates are increasingly being used as animal models in neuroscience research.However,efficient neuronal tracing techniques for labeling motor neurons and primary sensory afferents in the monkey spinal cord ...Nonhuman primates are increasingly being used as animal models in neuroscience research.However,efficient neuronal tracing techniques for labeling motor neurons and primary sensory afferents in the monkey spinal cord are lacking.Here,by injecting the cholera toxin B subunit into the sciatic nerve of a rhesus monkey,we successfully labeled the motor neurons and primary sensory afferents in the lumbar and sacralspinal cord.Labeled alpha motor neurons were located in lamina IX of the L6–S1 segments,which innervate both flexors and extensors.The labeled primary sensory afferents were mainly myelinated Aβfibers that terminated mostly in laminae I and II of the L4–L7 segments.Together with the labeled proprioceptive afferents,the primary sensory afferents formed excitatory synapses with multiple types of spinal neurons.In summary,our methods successfully traced neuronal connections in the monkey spinal cord and can be used in spinal cord studies when nonhuman primates are used.展开更多
Neural tube defects(NTDs)are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure.Although folate supplementation has been shown to mitigate the incidence of NTDs,some cases,often...Neural tube defects(NTDs)are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure.Although folate supplementation has been shown to mitigate the incidence of NTDs,some cases,often attributable to genetic factors,remain unpreventable.The SHROOM3 gene has been implicated in NTD cases that are unresponsive to folate supplementation;at present,however,the underlying mechanism remains unclear.Neural tube morphogenesis is a complex process involving the folding of the planar epithelium of the neural plate.To determine the role of SHROOM3 in early developmental morphogenesis,we established a neuroepithelial organoid culture system derived from cynomolgus monkeys to closely mimic the in vivo neural plate phase.Loss of SHROOM3 resulted in shorter neuroepithelial cells and smaller nuclei.These morphological changes were attributed to the insufficient recruitment of cytoskeletal proteins,namely fibrous actin(F-actin),myosin II,and phospho-myosin light chain(PMLC),to the apical side of the neuroepithelial cells.Notably,these defects were not rescued by folate supplementation.RNA sequencing revealed that differentially expressed genes were enriched in biological processes associated with cellular and organ morphogenesis.In summary,we established an authentic in vitro system to study NTDs and identified a novel mechanism for NTDs that are unresponsive to folate supplementation.展开更多
Urban environments expose animals to abundant anthropogenic materials and foods that facilitate foraging innovations in species with oppor-tunistic diets and high behavioral flexibility.Neophilia and exploration tende...Urban environments expose animals to abundant anthropogenic materials and foods that facilitate foraging innovations in species with oppor-tunistic diets and high behavioral flexibility.Neophilia and exploration tendency are believed to be important behavioral traits for animals thriving in urban environments.Vervet monkeys(Chlorocebus pygerythrus)are one of few primate species that have successfully adapted to urban environments,thus making them an ideal species to study these traits.Using a within-species cross-habitat approach,we compared neophilia and exploration of novel objects(jointly referred to as"object curiosity")between semi-urban,wild,and captive monkeys to shed light on the cognitive traits facilitating urban living.To measure"object curiosity:"we exposed monkeys to various types of novel stimuli and compared their approaches and explorative behavior.Our results revealed differences in the number of approaches and explorative behavior toward novel stimuli between the habitat types considered.Captive vervet monkeys were significantly more explorative than both semi-urban and wild troops,sug-gesting that positive experiences with humans and lack of predation,rather than exposure to human materials per se,influence object curiosity.Across habitats,juvenile males were the most explorative age-sex class.This is likely due to males being the dispersing sex and juveniles being more motivated to learn about their environment.Additionally,we found that items potentially associated with human food,elicited stronger explorative responses in semi-urban monkeys than non-food related objects,suggesting that their motivation to explore might be driven by"anthrophilia'that is,their experience of rewarding foraging on similar anthropogenic food sources.We conclude that varying levels of exposure to humans,predation and pre-exposure to human food packaging explain variation in"object curiosity"in our sample of vervet monkeys.展开更多
Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-gr...Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-grade AAV capsids have been shown to offer access to cor-tical projection neurons across different species and enable the manipulation of neural function in non-human primates(NHPs).Here,we report the development of a novel retro-grade AAV capsid,AAV-DJ8R,which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques.In addition,intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations.Moreover,AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen.These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.展开更多
DNA methylation plays a crucial role in environmental adaptations.Here,using whole-genome bisulfite sequencing,we generated comprehensive genome-wide DNA methylation profiles for the high-altitude Yunnan snub-nosed mo...DNA methylation plays a crucial role in environmental adaptations.Here,using whole-genome bisulfite sequencing,we generated comprehensive genome-wide DNA methylation profiles for the high-altitude Yunnan snub-nosed monkey(Rhinopithecus bieti)and the closely related golden snub-nosed monkey(R.roxellana).Our findings indicated a slight increase in overall DNA methylation levels in golden snub-nosed monkeys compared to Yunnan snub-nosed monkeys,suggesting a higher prevalence of hypermethylated genomic regions in the former.Comparative genomic methylation analysis demonstrated that genes associated with differentially methylated regions were involved in membrane fusion,vesicular formation and trafficking,hemoglobin function,cell cycle regulation,and neuronal differentiation.These results suggest that the high-altitude-related epigenetic modifications are extensive,involving a complete adaptation process from the inhibition of single Ca^(2+)channel proteins to multiple proteins collaboratively enhancing vesicular function or inhibiting cell differentiation and proliferation.Functional assays demonstrated that overexpression or down-regulation of candidate genes,such as SNX10,TIMELESS,and CACYBP,influenced cell viability under stress conditions.Overall,this research suggests that comparing DNA methylation across closely related species can identify novel candidate genomic regions and genes associated with local adaptations,thereby deepening our understanding of the mechanisms underlying environmental adaptations.展开更多
Coxsackievirus B3(CVB3)is the pathogen causing hand,foot and mouth disease(HFMD),which manifests across a spectrum of clinical severity from mild to severe.However,CVB3-infected mouse models mainly demonstrate viral m...Coxsackievirus B3(CVB3)is the pathogen causing hand,foot and mouth disease(HFMD),which manifests across a spectrum of clinical severity from mild to severe.However,CVB3-infected mouse models mainly demonstrate viral myocarditis and pancreatitis,failing to replicate human HFMD symptoms.Although several enteroviruses have been evaluated in Syrian hamsters and rhesus monkeys,there is no comprehensive data on CVB3.In this study,we have first tested the susceptibility of Syrian hamsters to CVB3 infection via different routes.The results showed that Syrian hamsters were successfully infected with CVB3 by intraperitoneal injection or nasal drip,leading to nasopharyngeal colonization,acute severe pathological injury,and typical HFMD symptoms.Notably,the nasal drip group exhibited a longer viral excretion cycle and more severe pathological damage.In the subsequent study,rhesus monkeys infected with CVB3 through nasal drips also presented signs of HFMD symptoms,viral excretion,serum antibody conversion,viral nucleic acids and antigens,and the specific organ damages,particularly in the heart.Surprisingly,there were no significant differences in myocardial enzyme levels,and the clinical symptoms resembled those often associated with common,mild infections.In summary,the study successfully developed severe Syrian hamsters and mild rhesus monkey models for CVB3-induced HFMD.These models could serve as a basis for understanding the disease pathogenesis,conducting pre-trial prevention and evaluation,and implementing post-exposure intervention.展开更多
The objective of this study was to set up a rhesus monkey model of polycystic ovary syndrome（PCOS）,which is globally prevalent among reproductive-aged human women,and to understand the reproductive traits of PCOS fe...The objective of this study was to set up a rhesus monkey model of polycystic ovary syndrome（PCOS）,which is globally prevalent among reproductive-aged human women,and to understand the reproductive traits of PCOS female monkeys.Six adult female rhesus monkeys aged 6-10 a,were divided into a PCOS group and a control group.The PCOS group were given two cycles of subcutaneous injections of propionic acid testosterone（PAT）,3.5 mg/kg body weight,on day 1,day 3,and day 5 of the menstrual cycle,respectively,and then given muscle injections of human chorionic gonadotropin（HCG）,350 IU/kg body wtight,on day 7,day 9,and day 11,respectively.Results showed that high levels of serum LH and T [（5.35±0.17） IU/L and（7.58±0.14） ng/mL,respectively],and a high ratio value of LH/FSH（5.35/1.30=4.12） were observed in the PCOS group.No significant differences were found in serum FSH,E2,and P in the PCOS group compared with those of the control.Polycystic ovaries in the PCOS monkeys were recorded by live ultrasound.The blastocysts rates of the PCOS vs.the control were 23.53% vs.66.67%,and there was a significant difference between the two groups.This study shows that PAT coupled with HCG can induce PCOS in rhesus monkeys in the short term.The reproductive features of PCOS monkeys were similar to those of PCOS patients.展开更多
Rhesus monkey embryonic stem（rES） cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,wh...Rhesus monkey embryonic stem（rES） cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,which are composed of cholangiocytes.However,little is known about the role of Notch signaling in cholangiocytic commitment of ES cells.We analyzed the effect of Notch signaling on the induction of cholangiocyte-like cells from rES cells.About 80% of definitive endoderm（DE） cells were generated from rES cells after treatment with activin A.After treatment with BMP4 and FGF1 on matrigel coated wells in serum-free medium,rES-derived DE gave rise to cholangiocyte-like cells by expression of cholangiocytic specific proteins（CK7,CK18,CK19,CK20,and OV-6） and genes（GSTPi,IB4,and HNF1β）.At the same time,expression of Notch 1 and Notch 2 mRNA were detected during cell differentiation,as well as their downstream target genes such as Hes 1 and Hes 5.Inhibition of the Notch signal pathway by L-685458 resulted in decreased expression of Notch and their downstream genes.In addition,the proportion of cholangiocyte-like cells declined from ～90% to ～20%.These results suggest that Notch signaling may play a critical role in cholangiocytic development from ES cells.展开更多
Objective To investigate the expression of nerve growth factor （NGF） and glial cell line-derived neurotrophic factor （GDNF） in monkeys of resuscitation after selective cerebral ultraprofound hypothermia and blood ...Objective To investigate the expression of nerve growth factor （NGF） and glial cell line-derived neurotrophic factor （GDNF） in monkeys of resuscitation after selective cerebral ultraprofound hypothermia and blood flow occlusion. Methods The monkeys were immediately removed brain after death in operation of group A （identical temperature perfusion group） and group B （ultraprofound hypothermia perfusion group）. Immunohistochemical technique was used to determine frontal cellular expression of NGF and GDNF. Statistics were analyzed by ANOVA analyses with significance level at P 〈 0.05. Results The expressions of NGF and GDNF in the group B were significantly higher than those in the group A （P 〈 0.05）. Conclusion NGF and GDNF increased significantly in the monkeys of resuscitation after selective cerebral ultraprofound hypothermia and blood flow occlusion. It may be a protective mechanism for neuron survival and neural function recovery.展开更多
Guizhou golden monkey(Rhinopithecus brelichi)was a unique,endangered and endemic primate species in Guizhou Province.It was an isolated population caused by habitat loss and fragmentation due to the human disturbance ...Guizhou golden monkey(Rhinopithecus brelichi)was a unique,endangered and endemic primate species in Guizhou Province.It was an isolated population caused by habitat loss and fragmentation due to the human disturbance in the recently 30 years in China,only distributed in Fanjingshan National Natural Reserve,Guizhou Province.To know the background with demonic population structure,we sequenced 867 bp of the mitochondrial DNA D-loop from 312 fresh fecal samples,results showed there 11 haplotypes among these samples,h was 0.517,πwas 0.00413.It indicated that this species had the lowest genetic diversity among four golden monkeys in China and need strengthen the conversation concern for this species immediately.展开更多
Rhesus monkey can not achieve natural delivery due to various reasons,and cesarean section becomes an important midwifery to get infant monkeys. After caesarean section,the pregnant monkey is weak and postoperative wo...Rhesus monkey can not achieve natural delivery due to various reasons,and cesarean section becomes an important midwifery to get infant monkeys. After caesarean section,the pregnant monkey is weak and postoperative wound pain,so it can not personally feed infant monkeys which must be artificially fed. Thus,establishing suitable feeding management program is very important for improving survival rate of infant rhesus monkey and maintaining good health. We summarized food preparation method for infant rhesus monkeys as well as temperature setting and light control,and established the nursing program for newborn infant monkey and daily management process for infant monkeys.展开更多
Gene editing in model organisms has provided critical insights into brain development and diseases. Here, we report the generation of a cynomolgus monkey （Macaca fascicularis） carrying MECP2 mutations using transcri...Gene editing in model organisms has provided critical insights into brain development and diseases. Here, we report the generation of a cynomolgus monkey （Macaca fascicularis） carrying MECP2 mutations using transcription activator-like effector nucleases （TALENs）-mediated gene targeting. After injecting TALENs mRNA into monkey zygotes achieved by in vitro fertilization and embryo transplantation into surrogate monkeys, we obtained one male newborn monkey with an MECP2 deletion caused by frame- shifting mutation in various tissues. The monkey carrying the MECP2 mutation failed to survive after birth, due to either the toxicity of TALENs or the critical requirement of MECP2 for neural development. The level of MeCP2 protein was essentially depleted in the monkey＇s brain. This study demonstrates the feasibility of introducing genetic mutations in non-human primates by site-specific gene-editing methods.展开更多
The purpose of this research was to study the pharmacokinetics and the bioavailability of recombinant human parathyroid hormone [rhPTH （1-34）] in Rhesus monkeys after single and multiple subcutaneous administration....The purpose of this research was to study the pharmacokinetics and the bioavailability of recombinant human parathyroid hormone [rhPTH （1-34）] in Rhesus monkeys after single and multiple subcutaneous administration. An immunoradiometric assay （IRMA） was used to determine the plasma drug concentration of rhFFH （1-34） after giving single dose of 10, 20 and 40 ug/kg and daily dose of 40 ug/kg for 7 d by subcutaneous administration, and intravenous injection of 20 ug/kg in Rhesus monkeys. The pharmacokinetic parameters were calculated by noncompartmental analysis. The drug plasma level quantitation range was from 0.027 to 2.22 ng/mL. The intra- and inter-assay precision （CV） of analysis were less than 15%, and the average recovery was about 93.0% ± 8.6% - 116.5% ± 14.0%. After subcutaneous administration of rhPTH（1-34） at dose of 10, 20 and 40 ug/kg, the average Tmax was 0.67, 0.5 and 0.83 h, Cmax were 1.85 ± 0.05, 3.23 ± 0.25 and 7.15 ± 1.19 ng/mL, the AUC（0-∞） were 3.4 ± 0.6, 10.7 ± 1.3 and 12.6 ± 1.5 ng/h/mL, and terminal-phase elimination T1/2 were 0.72 ± 0.10, 1.15 ± 0.10 and 1.03 ± 0.06 h, respectively. The absolute bioavailability of rhPTH （1-34） was 46.96% after subcutaneous administration of 20 ug/kg. There was no evidence of accumulation during systemic exposure of rhPTH （1-34） upon multiple dosing in Rhesus monkeys. The IRMA assay method provide reasonable sensitivity and specificity for the pharrnacokinetic study of rhPTH （1-34） after subcutaneous or intravenous administration in Rhesus monkeys. The pharmacokinetic characteristic of rhPTH （1-34） in monkeys shows linear relationship with the dose administered subcutaneously.展开更多
Go/NoGo tasks are a useful behavioral model in the study of cognitive neurosciences. The present developmental study is aimed at establishing a developmental protocol of Go/NoGo visual-discrimination tasks to investig...Go/NoGo tasks are a useful behavioral model in the study of cognitive neurosciences. The present developmental study is aimed at establishing a developmental protocol of Go/NoGo visual-discrimination tasks to investigate more cognitive process. We used two rhesus monkeys to test our procedures. Our results suggested that the monkeys quickly learned Go/NoGo visual-discrimination tasks, and performed NoGo tasks better and easier than Go tasks. Using this visual-discrimination task, we can easily study related cognitive neurosciences.展开更多
Aim： To examine the possible effect of heat treatment on expression of heat shock proteins （Hsps） 105, 70, and 60 in primary monkey Sertoli cells and to evaluate the possible signal pathways. Methods： Western blot...Aim： To examine the possible effect of heat treatment on expression of heat shock proteins （Hsps） 105, 70, and 60 in primary monkey Sertoli cells and to evaluate the possible signal pathways. Methods： Western blot analysis, realtime polymerase chain reaction （PCR）, and confocal immunohistochemistry were used to analyze mRNA and protein levels of the Hsps in response to 43~C treatment of Sertoli cells isolated from pubertal monkey testes. Results： Staining with Hoechst 33342 indicated Sertoli cells did not undergo apoptosis after heat treatment. Hspl05 was expressed in cytoplasm of untreated Sertoli cells. Both Hspl05 mRNA and protein levels were increased approximately 20-fold compared to those of the untreated controls at 12 h after heat treatment. Untreated Sertoli cells did not express Hsp70, but heat stress induced its expression in the cell cytoplasm. The time-course of changes in Hsp70 was similar to that of Hsp105. In contrast to Hsp105 and Hsp70, the change in Hsp60 expression was much less obvious. The protein level between 12 h and 48 h after heat treatment was only approximately 1.5-fold that of the untreated control. Extracellular regulated kinase （ERK） 1/2 inhibitor （U0126） or phosphoinositide kinase-3 （PI3K） inhibitor （LY294002） could partially block the response of Hspl05 and Hsp70 induced by heat treatment. Conclusion： These results indicate that the heat-induced expression of the three types of Hsp in monkey Sertoli cells might be regulated by ERK and/or PI3K signal pathways, but the profile of their expression is different, suggesting that they might have different regulatory functions in Sertoli cells.展开更多
文摘1 If you visit Lopburi,you might feel like you are on the set of Rise of the Planet of the Apes.While James Franco isnt wandering down the streets,the city is overrun with so many monkeys that you might wonder if theyre taking over.These arent just any monkeys;theyre lively,curious,and absolutely everywhere.
基金supported by the Key-Area Research and Development Program of Guangdong Province(No.2019B030335001)the STI2030-Major Projects(No.2021ZD0200900)+9 种基金the National Key Research and Development Program of China(Nos.2021YFF0702700 and 2018YFA0801403)the National Natural Science Foundation of China(Nos.81960422,32100801,82101241,82360226,82160923,82260929,82374425,and 32460194)the Strategic Priority Research Program of the Chinese Academy of Sciences(CAS)(No.XDB32060200)the STI2030-Major Projects(Nos.2022ZD0205200 and 2022ZD0212700)the Science and Technology Department of Yunnan Province(Nos.202305AH340006 and 202305AH340007)the Yunnan Fundamental Research Projects(Nos.202201AT070153 and 202201AT070139)the Science and Technology Project of Yunnan Province(No.202101AY070001-001)the Yunnan Revitalization Talent Support Program(No.YNWR-QNBJ2019-043)the CAS“Light of West China”Programthe Yunnan Revitalization Talents Support Plan。
文摘Complex brain diseases seriously endanger human health,and early diagnostic biomarkers and effective treatments are currently lacking.Due to ethical constraints on human research,establishing monkey models is crucial to address these issues.With the rapid development of technology,transgenic monkey models of a range of brain diseases,especially autism spectrum disorder(ASD),have been successfully established.However,to establish practical and effective brain disease models and subsequently apply them to disease mechanism and treatment studies,there is still a lack of a standard tool,i.e.,a system for collecting and analyzing the daily behaviors of brain disease model monkeys.Therefore,with the goal of undertaking a comprehensive and quantitative study of behavioral phenotypes,we established a standard daily behavior collection and analysis system,including behavioral data collection protocols and a monkey daily behavior ethogram(MDBE)for rhesus and cynomolgus monkeys,which are the most commonly used non-human primates in model construction.Then,we used ASD as an application example after referring to the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition,Text Revision(DSM-5-TR),which is widely used in clinical disease diagnosis to obtain ASD core clinical symptoms.We then established a sub-ethogram(ASD monkey core behavior ethogram(MCBE-ASD))specifically for quantitative assessment of the core clinical symptoms of an ASD monkey model based on MDBE.Subsequently,we demonstrated the high reproducibility of the system.
基金National Resources Center for Non Human PrimatesNational Key R&D Project of China,Grant/Award Number:2021YFF0702804。
文摘Background:Non-human primates(NPHs),such as rhesus macaques,cynomol-gus monkeys,and Assamese macaques,play a crucial role in biomedical research.However,baseline cytokine and electrolyte data for these three species,particularly data stratified by age and sex,are limited.Therefore,the aim of this study was to establish and analyze age-and sex-specific cytokine and electrolyte profiles in these three species.Methods:This study included 40 rhesus macaques(21 males,19 females),33 cyn-omolgus monkeys(17 males,16 females),and 45 Assamese macaques(25 males,20 females)classified by age(1-5 years,6-12 years,>13 years)and sex.The levels of 23 immune function indicators and 5 electrolyte indicators were measured.Results:Among the three monkey species,the levels of sCD40L,IL-18,MCP-1,MIP-1β,TGFa,K^(+),Na^(+),and Cl^(-)exhibited species-,sex-,and age-related differences.Comparison within the same species,sex had no significant impact on cytokine levels in NHPs but did affect electrolyte levels,particularly Cl^(-)and Na^(+)levels,in cynomol-gus monkeys and Assamese macaques.Electrolyte levels in NHPs were not affected by age,whereas the levels of certain cytokines,particularly sCD40L,GM-CSF,and IL-10,varied with age.The remaining 21 cytokines demonstrated no significant age-related changes.Conclusions:Significant variations in cytokine and electrolyte levels exist among dif-ferent monkey species,sexes,and age groups.This research provides valuable re-sources for NHP researchers and sets the stage for further exploring the impacts of sex and age on NHP physiology and immune function.
基金supported by the National Science and Technology Innovation 2030 Major Program(2021ZD0200900)National Key Research and Development Program of China(2022YFF0710901)+3 种基金National Natural Science Foundation of China(82021001,31825018)Biological Resources Program of the Chinese Academy of Sciences(KFJBRP-005)111 Project D18007a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘While viral infections can disturb the host gut microbiome,the dynamic alterations in microbial composition following infection remain poorly characterized.This study identified SRV-8-infected monkeys and classified them into five groups based on infection progression.16S rRNA amplicon sequencing revealed significant alterations in the relative and inferred absolute abundance of bacterial genera UCG-002,Agathobacter,Coprococcus,and Holdemanella during the early stage of SRV-8 infection,coinciding with provirus formation.These microbial shifts were accompanied by functional modifications in bacterial communities at the same stage.In contrast,ITS amplicon sequencing indicated no significant differences in fungal composition between healthy wild-type and SRV-8-infected monkeys.Spearman correlation analyses demonstrated close interactions between intestinal bacteria and fungi following SRV-8 infection.Additionally,SRV-8 seropositive groups exhibited significantly elevated mRNA expression levels of pro-inflammatory(TNF-α,IFN-γ,IL-1β,and IL-6)and anti-inflammatory(IL-10)cytokine genes,highlighting close associations between inflammatory cytokines and immune responses.Overall,these findings provide a comprehensive characterization of bacterial and fungal microbiota dynamics and inflammatory cytokine responses associated with SRV-8 infection,clarifying the pathobiological mechanisms underlying SRV-8 infection from the perspective of the gut microbiome.
基金supported by a grant from Ministry of Science and Technology China,No.2022ZD0204704(to WW)the National Natural Science Foundation of China,No.82301572(to XZ)the China Postdoctoral Science Foundation,No.2023M731202(to XZ)。
文摘Nonhuman primates are increasingly being used as animal models in neuroscience research.However,efficient neuronal tracing techniques for labeling motor neurons and primary sensory afferents in the monkey spinal cord are lacking.Here,by injecting the cholera toxin B subunit into the sciatic nerve of a rhesus monkey,we successfully labeled the motor neurons and primary sensory afferents in the lumbar and sacralspinal cord.Labeled alpha motor neurons were located in lamina IX of the L6–S1 segments,which innervate both flexors and extensors.The labeled primary sensory afferents were mainly myelinated Aβfibers that terminated mostly in laminae I and II of the L4–L7 segments.Together with the labeled proprioceptive afferents,the primary sensory afferents formed excitatory synapses with multiple types of spinal neurons.In summary,our methods successfully traced neuronal connections in the monkey spinal cord and can be used in spinal cord studies when nonhuman primates are used.
基金supported by the National Natural Science Foundation of China (81930121,82125008 to Y.C.C.)National Key Research and Development Program of China (2018YFA0107902 to Y.C.C.and 2018YFA0801403 to Z.B.W.)+1 种基金Major Basic Research Project of Science and Technology of Yunnan (202001BC070001 to Y.C.C.)Natural Science Foundation of Yunnan Province (202102AA100053 to Y.C.C.)。
文摘Neural tube defects(NTDs)are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure.Although folate supplementation has been shown to mitigate the incidence of NTDs,some cases,often attributable to genetic factors,remain unpreventable.The SHROOM3 gene has been implicated in NTD cases that are unresponsive to folate supplementation;at present,however,the underlying mechanism remains unclear.Neural tube morphogenesis is a complex process involving the folding of the planar epithelium of the neural plate.To determine the role of SHROOM3 in early developmental morphogenesis,we established a neuroepithelial organoid culture system derived from cynomolgus monkeys to closely mimic the in vivo neural plate phase.Loss of SHROOM3 resulted in shorter neuroepithelial cells and smaller nuclei.These morphological changes were attributed to the insufficient recruitment of cytoskeletal proteins,namely fibrous actin(F-actin),myosin II,and phospho-myosin light chain(PMLC),to the apical side of the neuroepithelial cells.Notably,these defects were not rescued by folate supplementation.RNA sequencing revealed that differentially expressed genes were enriched in biological processes associated with cellular and organ morphogenesis.In summary,we established an authentic in vitro system to study NTDs and identified a novel mechanism for NTDs that are unresponsive to folate supplementation.
基金the funding agencies behind this research,the Finnish KONE Foundation,the Wenner Gren Foundation,and the Swiss National Science Foundation(CRSK-3_220769)awarded to SE.
文摘Urban environments expose animals to abundant anthropogenic materials and foods that facilitate foraging innovations in species with oppor-tunistic diets and high behavioral flexibility.Neophilia and exploration tendency are believed to be important behavioral traits for animals thriving in urban environments.Vervet monkeys(Chlorocebus pygerythrus)are one of few primate species that have successfully adapted to urban environments,thus making them an ideal species to study these traits.Using a within-species cross-habitat approach,we compared neophilia and exploration of novel objects(jointly referred to as"object curiosity")between semi-urban,wild,and captive monkeys to shed light on the cognitive traits facilitating urban living.To measure"object curiosity:"we exposed monkeys to various types of novel stimuli and compared their approaches and explorative behavior.Our results revealed differences in the number of approaches and explorative behavior toward novel stimuli between the habitat types considered.Captive vervet monkeys were significantly more explorative than both semi-urban and wild troops,sug-gesting that positive experiences with humans and lack of predation,rather than exposure to human materials per se,influence object curiosity.Across habitats,juvenile males were the most explorative age-sex class.This is likely due to males being the dispersing sex and juveniles being more motivated to learn about their environment.Additionally,we found that items potentially associated with human food,elicited stronger explorative responses in semi-urban monkeys than non-food related objects,suggesting that their motivation to explore might be driven by"anthrophilia'that is,their experience of rewarding foraging on similar anthropogenic food sources.We conclude that varying levels of exposure to humans,predation and pre-exposure to human food packaging explain variation in"object curiosity"in our sample of vervet monkeys.
基金supported by Ministry of Science and Technology of China(2019YFA0903803 and 2018YFA0801404)National Natural Science Foundation of China(31871090,32000730,81961128019,and 81901397)+7 种基金Shenzhen Science and Technology Innovation Commission(JCYJ20190809171003698,JCYJ202103243001018,JCYJ20180507182505475,and JCYJ20180504165804015)Shenzhen Technological Research Center for Primate Translational Medicine(F-2021-Z99-504979)Youth Innovation Promotion Association(CAS 2017120)Chinese Academy of Sciences International Partnership Program(172644KYSB20170004)China Postdoctoral Science Foundation(2019M653115.)CAS Key Laboratory of Brain Connectome and Manipulation(2019DP173024)Guangdong Provincial Key Laboratory of Brain Connectome and Behavior(2017B030301017)International Science and Technology Cooperation Base of Guangdong Province(2019A050505008).
文摘Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-grade AAV capsids have been shown to offer access to cor-tical projection neurons across different species and enable the manipulation of neural function in non-human primates(NHPs).Here,we report the development of a novel retro-grade AAV capsid,AAV-DJ8R,which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques.In addition,intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations.Moreover,AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen.These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.
基金supported by the National Natural Science Foundation of China(32330015,31821001)Strategic Priority Research Program of the Chinese Academy of Sciences(XDB31000000)。
文摘DNA methylation plays a crucial role in environmental adaptations.Here,using whole-genome bisulfite sequencing,we generated comprehensive genome-wide DNA methylation profiles for the high-altitude Yunnan snub-nosed monkey(Rhinopithecus bieti)and the closely related golden snub-nosed monkey(R.roxellana).Our findings indicated a slight increase in overall DNA methylation levels in golden snub-nosed monkeys compared to Yunnan snub-nosed monkeys,suggesting a higher prevalence of hypermethylated genomic regions in the former.Comparative genomic methylation analysis demonstrated that genes associated with differentially methylated regions were involved in membrane fusion,vesicular formation and trafficking,hemoglobin function,cell cycle regulation,and neuronal differentiation.These results suggest that the high-altitude-related epigenetic modifications are extensive,involving a complete adaptation process from the inhibition of single Ca^(2+)channel proteins to multiple proteins collaboratively enhancing vesicular function or inhibiting cell differentiation and proliferation.Functional assays demonstrated that overexpression or down-regulation of candidate genes,such as SNX10,TIMELESS,and CACYBP,influenced cell viability under stress conditions.Overall,this research suggests that comparing DNA methylation across closely related species can identify novel candidate genomic regions and genes associated with local adaptations,thereby deepening our understanding of the mechanisms underlying environmental adaptations.
基金supported by several key projects,the Medical and Health Science and Technology Innovation Project of the Chinese Academy of Medical Sciences(CIFMS,2016-I2M-2-001)the National Resource Center for Non-Human Primates,Major Science and Technology Special Projects in Yunnan Province,Kunming Science and Technology Innovation and Service Capacity Enhancement Program Key Projects(2016-2-R-07674)+3 种基金the CAMS Innovation Fund for Medical Sciences(CIFMS,2018-I2M-3-002 and 2021-I2M-1-024)the National Key R&D Project of China(2021YFF0702804)Peking Union Medical College-Central University Basic Scientific Research Business Fee(Project number.:3332023079)Yunnan Province Applied Basic Research Special Project-General Project(project number:202401CF070048,202301AT070367).
文摘Coxsackievirus B3(CVB3)is the pathogen causing hand,foot and mouth disease(HFMD),which manifests across a spectrum of clinical severity from mild to severe.However,CVB3-infected mouse models mainly demonstrate viral myocarditis and pancreatitis,failing to replicate human HFMD symptoms.Although several enteroviruses have been evaluated in Syrian hamsters and rhesus monkeys,there is no comprehensive data on CVB3.In this study,we have first tested the susceptibility of Syrian hamsters to CVB3 infection via different routes.The results showed that Syrian hamsters were successfully infected with CVB3 by intraperitoneal injection or nasal drip,leading to nasopharyngeal colonization,acute severe pathological injury,and typical HFMD symptoms.Notably,the nasal drip group exhibited a longer viral excretion cycle and more severe pathological damage.In the subsequent study,rhesus monkeys infected with CVB3 through nasal drips also presented signs of HFMD symptoms,viral excretion,serum antibody conversion,viral nucleic acids and antigens,and the specific organ damages,particularly in the heart.Surprisingly,there were no significant differences in myocardial enzyme levels,and the clinical symptoms resembled those often associated with common,mild infections.In summary,the study successfully developed severe Syrian hamsters and mild rhesus monkey models for CVB3-induced HFMD.These models could serve as a basis for understanding the disease pathogenesis,conducting pre-trial prevention and evaluation,and implementing post-exposure intervention.
基金supported by Yunnan Key Laboratory of Animal Reproductive Biology(2010-03)
文摘The objective of this study was to set up a rhesus monkey model of polycystic ovary syndrome（PCOS）,which is globally prevalent among reproductive-aged human women,and to understand the reproductive traits of PCOS female monkeys.Six adult female rhesus monkeys aged 6-10 a,were divided into a PCOS group and a control group.The PCOS group were given two cycles of subcutaneous injections of propionic acid testosterone（PAT）,3.5 mg/kg body weight,on day 1,day 3,and day 5 of the menstrual cycle,respectively,and then given muscle injections of human chorionic gonadotropin（HCG）,350 IU/kg body wtight,on day 7,day 9,and day 11,respectively.Results showed that high levels of serum LH and T [（5.35±0.17） IU/L and（7.58±0.14） ng/mL,respectively],and a high ratio value of LH/FSH（5.35/1.30=4.12） were observed in the PCOS group.No significant differences were found in serum FSH,E2,and P in the PCOS group compared with those of the control.Polycystic ovaries in the PCOS monkeys were recorded by live ultrasound.The blastocysts rates of the PCOS vs.the control were 23.53% vs.66.67%,and there was a significant difference between the two groups.This study shows that PAT coupled with HCG can induce PCOS in rhesus monkeys in the short term.The reproductive features of PCOS monkeys were similar to those of PCOS patients.
基金supported by research grants from Zhejiang Natural Sciences Foundation of China (Y2110911 Y2080996)the National Key Technologies R&D Program of China (2007CB947701)
文摘Rhesus monkey embryonic stem（rES） cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,which are composed of cholangiocytes.However,little is known about the role of Notch signaling in cholangiocytic commitment of ES cells.We analyzed the effect of Notch signaling on the induction of cholangiocyte-like cells from rES cells.About 80% of definitive endoderm（DE） cells were generated from rES cells after treatment with activin A.After treatment with BMP4 and FGF1 on matrigel coated wells in serum-free medium,rES-derived DE gave rise to cholangiocyte-like cells by expression of cholangiocytic specific proteins（CK7,CK18,CK19,CK20,and OV-6） and genes（GSTPi,IB4,and HNF1β）.At the same time,expression of Notch 1 and Notch 2 mRNA were detected during cell differentiation,as well as their downstream target genes such as Hes 1 and Hes 5.Inhibition of the Notch signal pathway by L-685458 resulted in decreased expression of Notch and their downstream genes.In addition,the proportion of cholangiocyte-like cells declined from ～90% to ～20%.These results suggest that Notch signaling may play a critical role in cholangiocytic development from ES cells.
基金This work was supported by the Key Program of Natural Science Foundation of Yunnan Province, China （No. 2003C0010Z）.
文摘Objective To investigate the expression of nerve growth factor （NGF） and glial cell line-derived neurotrophic factor （GDNF） in monkeys of resuscitation after selective cerebral ultraprofound hypothermia and blood flow occlusion. Methods The monkeys were immediately removed brain after death in operation of group A （identical temperature perfusion group） and group B （ultraprofound hypothermia perfusion group）. Immunohistochemical technique was used to determine frontal cellular expression of NGF and GDNF. Statistics were analyzed by ANOVA analyses with significance level at P 〈 0.05. Results The expressions of NGF and GDNF in the group B were significantly higher than those in the group A （P 〈 0.05）. Conclusion NGF and GDNF increased significantly in the monkeys of resuscitation after selective cerebral ultraprofound hypothermia and blood flow occlusion. It may be a protective mechanism for neuron survival and neural function recovery.
文摘Guizhou golden monkey(Rhinopithecus brelichi)was a unique,endangered and endemic primate species in Guizhou Province.It was an isolated population caused by habitat loss and fragmentation due to the human disturbance in the recently 30 years in China,only distributed in Fanjingshan National Natural Reserve,Guizhou Province.To know the background with demonic population structure,we sequenced 867 bp of the mitochondrial DNA D-loop from 312 fresh fecal samples,results showed there 11 haplotypes among these samples,h was 0.517,πwas 0.00413.It indicated that this species had the lowest genetic diversity among four golden monkeys in China and need strengthen the conversation concern for this species immediately.
基金Supported by Yunnan Technology Innovation Talent Project(2014HB090)Kunming Science and Technology Planning Project(2014-04-A-S-02-3074)
文摘Rhesus monkey can not achieve natural delivery due to various reasons,and cesarean section becomes an important midwifery to get infant monkeys. After caesarean section,the pregnant monkey is weak and postoperative wound pain,so it can not personally feed infant monkeys which must be artificially fed. Thus,establishing suitable feeding management program is very important for improving survival rate of infant rhesus monkey and maintaining good health. We summarized food preparation method for infant rhesus monkeys as well as temperature setting and light control,and established the nursing program for newborn infant monkey and daily management process for infant monkeys.
基金supported by the National Basic Research Development Program of China (2011CBA00400 and 2011CB809102)the CAS Strategic Priority Research Program of China (XDB02050400)+2 种基金the National Key Technology R&D Program of China (2014BAI03B00)the CAS Hundreds of Talents Program of China (to Z.Q. and Q.S.)the National Science Foundation of China (91232712)
文摘Gene editing in model organisms has provided critical insights into brain development and diseases. Here, we report the generation of a cynomolgus monkey （Macaca fascicularis） carrying MECP2 mutations using transcription activator-like effector nucleases （TALENs）-mediated gene targeting. After injecting TALENs mRNA into monkey zygotes achieved by in vitro fertilization and embryo transplantation into surrogate monkeys, we obtained one male newborn monkey with an MECP2 deletion caused by frame- shifting mutation in various tissues. The monkey carrying the MECP2 mutation failed to survive after birth, due to either the toxicity of TALENs or the critical requirement of MECP2 for neural development. The level of MeCP2 protein was essentially depleted in the monkey＇s brain. This study demonstrates the feasibility of introducing genetic mutations in non-human primates by site-specific gene-editing methods.
文摘The purpose of this research was to study the pharmacokinetics and the bioavailability of recombinant human parathyroid hormone [rhPTH （1-34）] in Rhesus monkeys after single and multiple subcutaneous administration. An immunoradiometric assay （IRMA） was used to determine the plasma drug concentration of rhFFH （1-34） after giving single dose of 10, 20 and 40 ug/kg and daily dose of 40 ug/kg for 7 d by subcutaneous administration, and intravenous injection of 20 ug/kg in Rhesus monkeys. The pharmacokinetic parameters were calculated by noncompartmental analysis. The drug plasma level quantitation range was from 0.027 to 2.22 ng/mL. The intra- and inter-assay precision （CV） of analysis were less than 15%, and the average recovery was about 93.0% ± 8.6% - 116.5% ± 14.0%. After subcutaneous administration of rhPTH（1-34） at dose of 10, 20 and 40 ug/kg, the average Tmax was 0.67, 0.5 and 0.83 h, Cmax were 1.85 ± 0.05, 3.23 ± 0.25 and 7.15 ± 1.19 ng/mL, the AUC（0-∞） were 3.4 ± 0.6, 10.7 ± 1.3 and 12.6 ± 1.5 ng/h/mL, and terminal-phase elimination T1/2 were 0.72 ± 0.10, 1.15 ± 0.10 and 1.03 ± 0.06 h, respectively. The absolute bioavailability of rhPTH （1-34） was 46.96% after subcutaneous administration of 20 ug/kg. There was no evidence of accumulation during systemic exposure of rhPTH （1-34） upon multiple dosing in Rhesus monkeys. The IRMA assay method provide reasonable sensitivity and specificity for the pharrnacokinetic study of rhPTH （1-34） after subcutaneous or intravenous administration in Rhesus monkeys. The pharmacokinetic characteristic of rhPTH （1-34） in monkeys shows linear relationship with the dose administered subcutaneously.
基金Chinese Academy of Science Grants (KSCX2-SWfor M.Y.) , National Basic Research Programof China , Chinese NationalScience Foundation (30470553) Programof Chinese Academy of Science (KJCX1-01 , KJCX1-09)
文摘Go/NoGo tasks are a useful behavioral model in the study of cognitive neurosciences. The present developmental study is aimed at establishing a developmental protocol of Go/NoGo visual-discrimination tasks to investigate more cognitive process. We used two rhesus monkeys to test our procedures. Our results suggested that the monkeys quickly learned Go/NoGo visual-discrimination tasks, and performed NoGo tasks better and easier than Go tasks. Using this visual-discrimination task, we can easily study related cognitive neurosciences.
基金Acknowledgment This study was supported by the ＂973＂ project （No. 2006CB504001）, the Major Research Plan （No. 2006CB944001）, the CAS Innovation Project （KSCA2- YW-R-55）, the National Natural Science Foundation of China （No. 3061800530230190 30600311）, and the Beijing Natural Science Foundation （No. 5073032）.
文摘Aim： To examine the possible effect of heat treatment on expression of heat shock proteins （Hsps） 105, 70, and 60 in primary monkey Sertoli cells and to evaluate the possible signal pathways. Methods： Western blot analysis, realtime polymerase chain reaction （PCR）, and confocal immunohistochemistry were used to analyze mRNA and protein levels of the Hsps in response to 43~C treatment of Sertoli cells isolated from pubertal monkey testes. Results： Staining with Hoechst 33342 indicated Sertoli cells did not undergo apoptosis after heat treatment. Hspl05 was expressed in cytoplasm of untreated Sertoli cells. Both Hspl05 mRNA and protein levels were increased approximately 20-fold compared to those of the untreated controls at 12 h after heat treatment. Untreated Sertoli cells did not express Hsp70, but heat stress induced its expression in the cell cytoplasm. The time-course of changes in Hsp70 was similar to that of Hsp105. In contrast to Hsp105 and Hsp70, the change in Hsp60 expression was much less obvious. The protein level between 12 h and 48 h after heat treatment was only approximately 1.5-fold that of the untreated control. Extracellular regulated kinase （ERK） 1/2 inhibitor （U0126） or phosphoinositide kinase-3 （PI3K） inhibitor （LY294002） could partially block the response of Hspl05 and Hsp70 induced by heat treatment. Conclusion： These results indicate that the heat-induced expression of the three types of Hsp in monkey Sertoli cells might be regulated by ERK and/or PI3K signal pathways, but the profile of their expression is different, suggesting that they might have different regulatory functions in Sertoli cells.
