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
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
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
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
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
For group-living primates, the information on postconflict management is crucial for understanding primate competition and cooperation. However, such information is poorly known for snub-nosed monkeys, especially for ...For group-living primates, the information on postconflict management is crucial for understanding primate competition and cooperation. However, such information is poorly known for snub-nosed monkeys, especially for wild populations. In this study, from September 2007 to June 2008, we investigated postconflict behavior among adult females Sichuan snub-nosed monkeys Rhinopithecus roxellana within one-male units in a wild, provisioned group in the Qinling Mountains of China by means of the time-rule method and the PC-MC method. We obtained a total of 81 PC-MC pairs and each individual was involved in only 0.004 aggressive behavior per observation hour. The first affiliative behavior was more likely to occur within the first minute after a conflict. The postconflict affiliative behaviors most often seen were contact-sit, embrace and grooming. The affiliative contacts between adult females occur due to selective attraction, i.e. reconciliation. The pattern of postconflict affiliation demonstrates that the R. roxellana belongs to a tolerant species.展开更多
Hypobaric hypoxia(HH)exposure can cause serious brain injury as well as life-threatening cerebral edema in severe cases.Previous studies on the mechanisms of HH-induced brain injury have been conducted primarily using...Hypobaric hypoxia(HH)exposure can cause serious brain injury as well as life-threatening cerebral edema in severe cases.Previous studies on the mechanisms of HH-induced brain injury have been conducted primarily using non-primate animal models that are genetically distant to humans,thus hindering the development of disease treatment.Here,we report that cynomolgus monkeys(Macaca fascicularis)exposed to acute HH developed human-like HH syndrome involving severe brain injury and abnormal behavior.Transcriptome profiling of white blood cells and brain tissue from monkeys exposed to increasing altitude revealed the central role of the HIF-1 and other novel signaling pathways,such as the vitamin D receptor(VDR)signaling pathway,in co-regulating HH-induced inflammation processes.We also observed profound transcriptomic alterations in brains after exposure to acute HH,including the activation of angiogenesis and impairment of aerobic respiration and protein folding processes,which likely underlie the pathological effects of HH-induced brain injury.Administration of progesterone(PROG)and steroid neuroprotectant 5α-androst-3β,5,6β-triol(TRIOL)significantly attenuated brain injuries and rescued the transcriptomic changes induced by acute HH.Functional investigation of the affected genes suggested that these two neuroprotectants protect the brain by targeting different pathways,with PROG enhancing erythropoiesis and TRIOL suppressing glutamate-induced excitotoxicity.Thus,this study advances our understanding of the pathology induced by acute HH and provides potential compounds for the development of neuroprotectant drugs for therapeutic treatment.展开更多
In vitro culture of spermatogonial stem cells （SSCs） has generally been performed using two-dimensional （2D） culture systems; however, such cultures have not led to the development of complete spermatogenesis. It ...In vitro culture of spermatogonial stem cells （SSCs） has generally been performed using two-dimensional （2D） culture systems; however, such cultures have not led to the development of complete spermatogenesis. It seems that 2D systems do not replicate optimal conditions of the seminiferous tubules （including those generated by the SSC niche） and necessary for spermatogenesis. Recently, one of our laboratories has been able to induce proliferation and differentiation of mouse testicular germ cells to meiotic and postmeiotic stages including generation of sperm in a 3D soft agar culture system （SACS） and a 3D methylcellulose culture system （MCS）. It was suggested that SACS and MCS form a special 3D microenvironment that mimics germ cell niche formation in the seminiferous tubules, and thus permits mouse spermatogenesis in vitro. In this review, we （1） provide a brief overview of the differences in spermatogenesis in rodents and primates, （2） summarize data related to attempts to generate sperm in vitro, （3） report for the first time formation of colonies/clusters of cells and differentiation of meiotic （expression of CREM-1） and postmeiotic （expression of acrosin） germ cells from undifferentiated spermatogonia isolated from the testis of prepubertal rhesus monkeys and cultured in SACS and MCS, and （4） indicate research needed to optimize 3D systems for in vitro primate spermatogenesis and for possible future application to man.展开更多
Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technolo...Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technologies are based on these systems. Although many viral vectors are used in rodents, their full application has been limited in non-human primates. To identify viral vectors that can stably and effectively express exogenous genes within non- human primates, eleven commonly used recombinant adeno-associated viral and lentiviral vectors, each carrying a gene to express green or red fluorescence, were injected into the parietal cortex of four rhesus monkeys. The expression of fluorescent cells was used to quantify transfection efficiency. Histological results revealed that recombinant adeno-associated viral vectors, especially the serotype 2/9 coupled with the cytomegalovirus, human synapsin I, or Ca2~/calmodulin-dependent protein kinase II promoters, and lentiviral vector coupled with the human ubiquitin C promoter, induced higher expression of fluorescent cells, representing high transfection efficiency. This is the first comparison of transfection efficiencies of different viral vectors carrying different promoters and serotypes in non-human primates （NHPs）. These results can be used as an aid to select optimal vectors to transfer exogenous genes into the central nervous system of non-human primates.展开更多
Aim： To assess the spatiotemporal changes in the expression of extracellular signal-regulated kinases 1 and 2 （ERK1/ 2）, c-Jun N-terminal kinases （JNK） and p38 mitogen-activated protein kinases （MAPK） in respon...Aim： To assess the spatiotemporal changes in the expression of extracellular signal-regulated kinases 1 and 2 （ERK1/ 2）, c-Jun N-terminal kinases （JNK） and p38 mitogen-activated protein kinases （MAPK） in response to heat stress in the cryptorchid testis, and to investigate a possible relation to Sertoli cell dedifferentiation. Methods： Immunohistochemistry and western blot were used to examine the expression and activation of ERK1/2, p38 and JNK in the cryptorchid testis at various stages after experimental cryptorchidism. Results： The abdominal temperature did not obviously change the total ERK1/2 expression but significantly activated phospho-ERK1/2 in the Sertoli cells of the cryptorchid testis. Heat stress increased total JNK expression in the Sertoli cells of the cryptorchid testis but did not activate phospho-JNK. Neither total p38 nor phospho-p38 was induced by heat stress in the Sertoli cells of the cryptorchid testis. Changes in the spatiotemporal expression of cytokeratin 18 （CK18）, a marker of immature or undifferentiated Sertoli cells, were induced in the cryptorchid testis in a pattern similar to the activation of ERK1/2. Condusion： The activation of ERK1/2 in the testis may be related to dedifferentiation of Sertoli cells under heat stress induced by experimental cryptorchidism.展开更多
Dear Editor,Since the global eradication of smallpox in the 1980s,monkeypox has become the most severe infectious disease caused by pathogenic orthopoxvirus(OPXV)infection.Monkeypox virus(MPXV)is the causative pathoge...Dear Editor,Since the global eradication of smallpox in the 1980s,monkeypox has become the most severe infectious disease caused by pathogenic orthopoxvirus(OPXV)infection.Monkeypox virus(MPXV)is the causative pathogen of monkeypox and was classified as a category of human infectious pathogen by theMinistry of Health of the People's Republic ofChina in 2006.Monkeypox has traditionally been endemic in West and Central Africa since it was discovered nearly 70 years ago(Breman et al.,1980).However,beginning in early May 2022,a sudden outbreak of monkeypox has taken place and remains ongoing in more than 100 nonendemic countries and territories located at Europe,Americas,and Asia(https://worldhealthorg.shinyapps.io/mpx_global/).展开更多
Whether direct manipulation of Parkinson’s disease(PD)risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue.Here,we used an adeno-associated virus serotype 9(AAV9)-deliver...Whether direct manipulation of Parkinson’s disease(PD)risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue.Here,we used an adeno-associated virus serotype 9(AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras(SNs)of two monkey groups:an old group and a middle-aged group.After the operation,the old group exhibited all the classic PD symptoms,including bradykinesia,tremor,and postural instability,accompanied by key pathological hallmarks of PD,such as severe nigral dopaminergic neuron loss(>64%)and evidentα-synuclein pathology in the gene-edited SN.In contrast,the phenotype of their middle-aged counterparts,which also showed clear PD symptoms and pathological hallmarks,were less severe.In addition to the higher final total PD scores and more severe pathological changes,the old group were also more susceptible to gene editing by showing a faster process of PD progression.These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys.Taken together,this system can effectively develop a large number of genetically-edited PD monkeys in a short time(6–10 months),and thus provides a practical transgenic monkey model for future PD studies.展开更多
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.
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 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.
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.
基金The Natural Science Foundation of China (No.30970444, No. 30770375, No. 30630016)the Cosmo Oil Eco Card Fund of Japan (2005-2010) support
文摘For group-living primates, the information on postconflict management is crucial for understanding primate competition and cooperation. However, such information is poorly known for snub-nosed monkeys, especially for wild populations. In this study, from September 2007 to June 2008, we investigated postconflict behavior among adult females Sichuan snub-nosed monkeys Rhinopithecus roxellana within one-male units in a wild, provisioned group in the Qinling Mountains of China by means of the time-rule method and the PC-MC method. We obtained a total of 81 PC-MC pairs and each individual was involved in only 0.004 aggressive behavior per observation hour. The first affiliative behavior was more likely to occur within the first minute after a conflict. The postconflict affiliative behaviors most often seen were contact-sit, embrace and grooming. The affiliative contacts between adult females occur due to selective attraction, i.e. reconciliation. The pattern of postconflict affiliation demonstrates that the R. roxellana belongs to a tolerant species.
基金supported by the National Natural Science Foundation of China(81773711)to W.Y.Strategic Priority Research Program of the Chinese Academy of Sciences(XDB13000000)+6 种基金Lundbeck Foundation Grant(R190-2014-2827)Carlsberg Foundation Grant(CF16-0663)to G.J.Z.Science and Technology Program of Guangzhou,China(201704020103)to W.Y.Introduction of Innovative R&D Team Program of Guangdong Province(2013Y104)Leading Talent Project in Science and Technology of Guangzhou Development District(2019-L002)National Major Scientific and Technological Special Project for“Significant New Drugs Development”(2016ZX09101026)to S.Z.L.Key Projects of the Military Science and Technology PLA(AWS14C007 and AWS16J023)to Y.Q.G
文摘Hypobaric hypoxia(HH)exposure can cause serious brain injury as well as life-threatening cerebral edema in severe cases.Previous studies on the mechanisms of HH-induced brain injury have been conducted primarily using non-primate animal models that are genetically distant to humans,thus hindering the development of disease treatment.Here,we report that cynomolgus monkeys(Macaca fascicularis)exposed to acute HH developed human-like HH syndrome involving severe brain injury and abnormal behavior.Transcriptome profiling of white blood cells and brain tissue from monkeys exposed to increasing altitude revealed the central role of the HIF-1 and other novel signaling pathways,such as the vitamin D receptor(VDR)signaling pathway,in co-regulating HH-induced inflammation processes.We also observed profound transcriptomic alterations in brains after exposure to acute HH,including the activation of angiogenesis and impairment of aerobic respiration and protein folding processes,which likely underlie the pathological effects of HH-induced brain injury.Administration of progesterone(PROG)and steroid neuroprotectant 5α-androst-3β,5,6β-triol(TRIOL)significantly attenuated brain injuries and rescued the transcriptomic changes induced by acute HH.Functional investigation of the affected genes suggested that these two neuroprotectants protect the brain by targeting different pathways,with PROG enhancing erythropoiesis and TRIOL suppressing glutamate-induced excitotoxicity.Thus,this study advances our understanding of the pathology induced by acute HH and provides potential compounds for the development of neuroprotectant drugs for therapeutic treatment.
基金Supported by National High Technology Research and Development Program of China 863 Programs No.2006AA02A141 and No.2012AA020505the Medical Research Fund of Guangdong Province No.2009164
文摘AIM: To evaluate a hybrid bioartificial liver support system (HBALSS) in cynomolgus monkeys with acute liver failure.
文摘In vitro culture of spermatogonial stem cells （SSCs） has generally been performed using two-dimensional （2D） culture systems; however, such cultures have not led to the development of complete spermatogenesis. It seems that 2D systems do not replicate optimal conditions of the seminiferous tubules （including those generated by the SSC niche） and necessary for spermatogenesis. Recently, one of our laboratories has been able to induce proliferation and differentiation of mouse testicular germ cells to meiotic and postmeiotic stages including generation of sperm in a 3D soft agar culture system （SACS） and a 3D methylcellulose culture system （MCS）. It was suggested that SACS and MCS form a special 3D microenvironment that mimics germ cell niche formation in the seminiferous tubules, and thus permits mouse spermatogenesis in vitro. In this review, we （1） provide a brief overview of the differences in spermatogenesis in rodents and primates, （2） summarize data related to attempts to generate sperm in vitro, （3） report for the first time formation of colonies/clusters of cells and differentiation of meiotic （expression of CREM-1） and postmeiotic （expression of acrosin） germ cells from undifferentiated spermatogonia isolated from the testis of prepubertal rhesus monkeys and cultured in SACS and MCS, and （4） indicate research needed to optimize 3D systems for in vitro primate spermatogenesis and for possible future application to man.
基金supported by the National Program on Key Basic Research Project(973 Programs 2015CB755605)the National Natural Science Foundation of China(81471312)
文摘Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technologies are based on these systems. Although many viral vectors are used in rodents, their full application has been limited in non-human primates. To identify viral vectors that can stably and effectively express exogenous genes within non- human primates, eleven commonly used recombinant adeno-associated viral and lentiviral vectors, each carrying a gene to express green or red fluorescence, were injected into the parietal cortex of four rhesus monkeys. The expression of fluorescent cells was used to quantify transfection efficiency. Histological results revealed that recombinant adeno-associated viral vectors, especially the serotype 2/9 coupled with the cytomegalovirus, human synapsin I, or Ca2~/calmodulin-dependent protein kinase II promoters, and lentiviral vector coupled with the human ubiquitin C promoter, induced higher expression of fluorescent cells, representing high transfection efficiency. This is the first comparison of transfection efficiencies of different viral vectors carrying different promoters and serotypes in non-human primates （NHPs）. These results can be used as an aid to select optimal vectors to transfer exogenous genes into the central nervous system of non-human primates.
基金Acknowledgment This study was supported by the National Natural Science Foundation of China （30230190）, the National Basic Science Research and Development Project （973） （G1999055901） and the Chinese Academy of Sciences （CAS） Knowledge Innovation Program （KSCX-2-SW-201）.
文摘Aim： To assess the spatiotemporal changes in the expression of extracellular signal-regulated kinases 1 and 2 （ERK1/ 2）, c-Jun N-terminal kinases （JNK） and p38 mitogen-activated protein kinases （MAPK） in response to heat stress in the cryptorchid testis, and to investigate a possible relation to Sertoli cell dedifferentiation. Methods： Immunohistochemistry and western blot were used to examine the expression and activation of ERK1/2, p38 and JNK in the cryptorchid testis at various stages after experimental cryptorchidism. Results： The abdominal temperature did not obviously change the total ERK1/2 expression but significantly activated phospho-ERK1/2 in the Sertoli cells of the cryptorchid testis. Heat stress increased total JNK expression in the Sertoli cells of the cryptorchid testis but did not activate phospho-JNK. Neither total p38 nor phospho-p38 was induced by heat stress in the Sertoli cells of the cryptorchid testis. Changes in the spatiotemporal expression of cytokeratin 18 （CK18）, a marker of immature or undifferentiated Sertoli cells, were induced in the cryptorchid testis in a pattern similar to the activation of ERK1/2. Condusion： The activation of ERK1/2 in the testis may be related to dedifferentiation of Sertoli cells under heat stress induced by experimental cryptorchidism.
基金supported by the National Science and Technology Major Project of China(2018ZX10711001-006).
文摘Dear Editor,Since the global eradication of smallpox in the 1980s,monkeypox has become the most severe infectious disease caused by pathogenic orthopoxvirus(OPXV)infection.Monkeypox virus(MPXV)is the causative pathogen of monkeypox and was classified as a category of human infectious pathogen by theMinistry of Health of the People's Republic ofChina in 2006.Monkeypox has traditionally been endemic in West and Central Africa since it was discovered nearly 70 years ago(Breman et al.,1980).However,beginning in early May 2022,a sudden outbreak of monkeypox has taken place and remains ongoing in more than 100 nonendemic countries and territories located at Europe,Americas,and Asia(https://worldhealthorg.shinyapps.io/mpx_global/).
基金This work was supported by the National Key R&D Program of China(2018YFA0801403)the Key-Area Research and Development Program of Guangdong Province(2019B030335001)+6 种基金the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB32060200)the National Program for Key Basic Research Projects(973 Program:2015CB755605)the National Natural Science Foundation of China(81471312,81771387,81460352,81500983,31700897,31700910,31800901,31625013,and 91732302)the Applied Basic Research Programs of Science and Technology Commission Foundation of Yunnan Province(2017FB109,2018FB052,2018FB053,2019FA007,and 202001AT070130)Chinese Academy of Sciences"Light of West China"Program,Shanghai Brain-Intelligence Project from Science and Technology Commission of Shanghai Municipality(16JC1420501)Shanghai Municipal Science and Technology Major Project(2018SHZDZX05)Open Large Infrastructure Research of Chinese Academy of Sciences,and China Postdoctoral Science Foundation(2018M631105).
文摘Whether direct manipulation of Parkinson’s disease(PD)risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue.Here,we used an adeno-associated virus serotype 9(AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras(SNs)of two monkey groups:an old group and a middle-aged group.After the operation,the old group exhibited all the classic PD symptoms,including bradykinesia,tremor,and postural instability,accompanied by key pathological hallmarks of PD,such as severe nigral dopaminergic neuron loss(>64%)and evidentα-synuclein pathology in the gene-edited SN.In contrast,the phenotype of their middle-aged counterparts,which also showed clear PD symptoms and pathological hallmarks,were less severe.In addition to the higher final total PD scores and more severe pathological changes,the old group were also more susceptible to gene editing by showing a faster process of PD progression.These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys.Taken together,this system can effectively develop a large number of genetically-edited PD monkeys in a short time(6–10 months),and thus provides a practical transgenic monkey model for future PD studies.
