New aspects of a small GTPase RAB35 in brain development and function

In eukaryotic cells,organelles in the secretory,lysosomal,and endocytic pathways actively exchange biological materials with each other through intracellular membrane trafficking,which is the process of transporting the cargo of proteins,lipids,and other molecules to appropriate compartments via transport vesicles or intermediates.These processes are strictly regulated by various small GTPases such as the RAS-like in rat brain(RAB)protein family,which is the largest subfamily of the RAS superfamily.Dysfunction of membrane trafficking affects tissue homeostasis and leads to a wide range of diseases,including neurological disorders and neurodegenerative diseases.Therefore,it is important to understand the physiological and pathological roles of RAB proteins in brain function.RAB35,a member of the RAB family,is an evolutionarily conserved protein in metazoans.A wide range of studies using cultured mammalian cells and model organisms have revealed that RAB35 mediates various processes such as cytokinesis,endocytic recycling,actin bundling,and cell migration.RAB35 is also involved in neurite outgrowth and turnover of synaptic vesicles.We generated brain-specific Rab35 knockout mice to study the physiological roles of RAB35 in brain development and function.These mice exhibited defects in anxiety-related behaviors and spatial memory.Strikingly,RAB35 is required for the precise positioning of pyramidal neurons during hippocampal development,and thereby for normal hippocampal lamination.In contrast,layer formation in the cerebral cortex occurred superficially,even in the absence of RAB35,suggesting a predominant role for RAB35 in hippocampal development rather than in cerebral cortex development.Recent studies have suggested an association between RAB35 and neurodegenerative diseases,including Parkinson's disease and Alzheimer's disease.In this review,we provide an overview of the current understanding of subcellular functions of RAB35.We also provide insights into the physiological role of RAB35 in mammalian brain development and function,and discuss the involvement of RAB35 dysfunction in neurodegenerative diseases.

Establishment of human cerebral organoid systems to model early neural development and assess the central neurotoxicity of environmental toxins

Human brain development is a complex process,and animal models often have significant limitations.To address this,researchers have developed pluripotent stem cell-derived three-dimensional structures,known as brain-like organoids,to more accurately model early human brain development and disease.To enable more consistent and intuitive reproduction of early brain development,in this study,we incorporated forebrain organoid culture technology into the traditional unguided method of brain organoid culture.This involved embedding organoids in matrigel for only 7 days during the rapid expansion phase of the neural epithelium and then removing them from the matrigel for further cultivation,resulting in a new type of human brain organoid system.This cerebral organoid system replicated the temporospatial characteristics of early human brain development,including neuroepithelium derivation,neural progenitor cell production and maintenance,neuron differentiation and migration,and cortical layer patterning and formation,providing more consistent and reproducible organoids for developmental modeling and toxicology testing.As a proof of concept,we applied the heavy metal cadmium to this newly improved organoid system to test whether it could be used to evaluate the neurotoxicity of environmental toxins.Brain organoids exposed to cadmium for 7 or 14 days manifested severe damage and abnormalities in their neurodevelopmental patterns,including bursts of cortical cell death and premature differentiation.Cadmium exposure caused progressive depletion of neural progenitor cells and loss of organoid integrity,accompanied by compensatory cell proliferation at ectopic locations.The convenience,flexibility,and controllability of this newly developed organoid platform make it a powerful and affordable alternative to animal models for use in neurodevelopmental,neurological,and neurotoxicological studies.

Against all odds:The road to success in the development of human immune reconstitution mice

The mouse genome has a high degree of homology with the human genome,and its physiological,biochemical,and developmental regulation mechanisms are similar to those of humans;therefore,mice are widely used as experimental animals.However,it is undeniable that interspecies differences between humans and mice can lead to experimental errors.The differences in the immune system have become an impor-tant factor limiting current immunological research.The application of immunodefi-cient mice provides a possible solution to these problems.By transplanting human immune cells or tissues,such as peripheral blood mononuclear cells or hematopoietic stem cells,into immunodeficient mice,a human immune system can be reconstituted in the mouse body,and the engrafted immune cells can elicit human-specific immune responses.Researchers have been actively exploring the development and differen-tiation conditions of host recipient animals and grafts in order to achieve better im-mune reconstitution.Through genetic engineering methods,immunodeficient mice can be further modified to provide a favorable developmental and differentiation microenvironment for the grafts.From initially only being able to reconstruct single T lymphocyte lineages,it is now possible to reconstruct lymphoid and myeloid cells,providing important research tools for immunology-related studies.In this review,we compare the differences in immune systems of humans and mice,describe the devel-opment history of human immune reconstitution from the perspectives of immuno-deficient mice and grafts,and discuss the latest advances in enhancing the efficiency of human immune cell reconstitution,aiming to provide important references for im-munological related researches.

Parallel Event of UNHRC50 Held in Beijing:Climate Change & Sustainable Development of Human Beings

On July 6,2022,a parallel event,organized during the 50th session of the United Nations Human Rights Council(HRC50),was co-hosted by Beijing NGO Network for International Exchanges(BNIE),China Association for NGO Cooperation(CANGO)and Beijing International Peace Culture Foundation(IPCF).The subject of this meeting focused on addressing climate change and promoting sustainable development of human beings.Experts,scholars and representatives of international organizations from relevant institutions at home and abroad had in-depth exchanges and discussions,making NGOs’contribution to international affairs and global ecological environment governance.

Study on the Epistemology Development of the Correlation between Human Beings and Nature

This study aimed to alert people that the harmony between human beings and nature has already become an urgent problem. All people are responsible for maintaining the harmony, which is the also the inexorable trend of historical development. Reviewing the evolution history of human kind, the relationship between human beings and nature from harmony to imbalance, from imbalance to a new harmony was analyzed from the macro perspective, and it was pointed out that the present society is in the transitional phase from imbalance to a new harmony and also a phase that people are consciously coordinating their relationship with nature. It was found through the study that a harmonious society relies on the harmony between human beings and nature, and the latter is a necessary condition for a real harmonious society, however, people always forget such a principle. The imbalanced relationship between human beings and nature has seriously influenced many aspects of social harmony, and become instable factors in China and international society, even threatened the development and survival of human beings. Thus, all people should make joint efforts in creating win-win conditions for the development between human beings and nature, and ensuring offspring a healthy and green earth.

On All-round Development of Human Rights in China

In October 2003, the Third Plenary Session of the Sixteenth Central Committee of the Communist Party of China （CPC） called for adherence to ＂the outlook toward development based on the fundamental interests of the people, toward an all-round, balanced and sustainable development.＂ The session also proposed to the National People＇s Congress （NPC） that respect for and protection of human rights be written into the country＇s Constitution. And accordingly, the NPC adopted a decision to revise the Constitution the following year. The call and the proposal were inter-related. The call sets the guideline, the theoretical basis and the orientation for the development of human rights in China. The pro- posal subsequently resulted in a revision of-the Constitution, the fundamental state law, which came as a legal guarantee for respect for and protection of human rights. Then, the ＂task of promoting an all-round development of the human rights cause＂ was written down into the country＇s Eleventh Five-Year Program for National Economic and Social Development （2006-2010）, which was approved by the NPC in March 2006. This was the first time that the development of human rights was written into a national program for social development. This means that an even closer link has been forged between China＇s human rights cause on one hand and the country＇s political, economic, cultural and social welfare undertakings on the other, thus preparing China＇s human rights cause for faster and sustainable development. We are now safe to predict that in the future, there will be even more and better rights the Chinese people are able to enjoy.

The Program of Action for All-round Development of China's Human Rights Cause on the New Journey in the New Era

The 20thNational Congress of the Communist Party of China was an epoch-making event in the history of China’s human rights development.Founded a century ago under the banner of“fighting for democracy and human rights”,the Communist Party of China has now embarked on a new journey for the development of human rights cause in a new era.

BMPRⅡ^(+)neural precursor cells isolated and characterized from organotypic neurospheres:an in vitro model of human fetal spinal cord development

Roof plate secretion of bone morphogenetic proteins(BMPs)directs the cellular fate of sensory neurons during spinal cord development,including the formation of the ascending sensory columns,though their biology is not well understood.Type-ⅡBMP receptor(BMPRⅡ),the cognate receptor,is expressed by neural precursor cells during embryogenesis;however,an in vitro method of enriching BMPRⅡ^(+)human neural precursor cells(hNPCs)from the fetal spinal cord is absent.Immunofluorescence was undertaken on intact second-trimester human fetal spinal cord using antibodies to BMPRⅡand leukemia inhibitory factor(LIF).Regions of highest BMPRⅡ^(+)immunofluorescence localized to sensory columns.Parenchymal and meningeal-associated BMPRⅡ^(+)vascular cells were identified in both intact fetal spinal cord and cortex by co-positivity with vascular lineage markers,CD34/CD39.LIF immunostaining identified a population of somas concentrated in dorsal and ventral horn interneurons,mirroring the expression of LIF receptor/CD118.A combination of LIF supplementation and high-density culture maintained culture growth beyond 10 passages,while synergistically increasing the proportion of neurospheres with a stratified,cytoarchitecture.These neurospheres were characterized by BMPRⅡ^(+)/MAP2ab^(+/–)/βⅢ-tubulin^(+)/nestin^(–)/vimentin^(–)/GFAP^(–)/NeuN^(–)surface hNPCs surrounding a heterogeneous core ofβⅢ-tubulin^(+)/nestin^(+)/vimentin^(+)/GFAP^(+)/MAP2ab^(–)/NeuN^(–)multipotent precursors.Dissociated cultures from tripotential neurospheres contained neuronal(βⅢ-tubulin^(+)),astrocytic(GFAP+),and oligodendrocytic(O4+)lineage cells.Fluorescence-activated cell sorting-sorted BMPRⅡ^(+)hNPCs were MAP2ab^(+/–)/βⅢ-tubulin^(+)/GFAP^(–)/O4^(–)in culture.This is the first isolation of BMPRⅡ^(+)hNPCs identified and characterized in human fetal spinal cords.Our data show that LIF combines synergistically with high-density reaggregate cultures to support the organotypic reorganization of neurospheres,characterized by surface BMPRⅡ^(+)hNPCs.Our study has provided a new methodology for an in vitro model capable of amplifying human fetal spinal cord cell numbers for>10 passages.Investigations of the role BMPRⅡplays in spinal cord development have primarily relied upon mouse and rat models,with interpolations to human development being derived through inference.Because of significant species differences between murine biology and human,including anatomical dissimilarities in central nervous system(CNS)structure,the findings made in murine models cannot be presumed to apply to human spinal cord development.For these reasons,our human in vitro model offers a novel tool to better understand neurodevelopmental pathways,including BMP signaling,as well as spinal cord injury research and testing drug therapies.

Treatment of spinal cord injury with biomaterials and stem cell therapy in non-human primates and humans

Spinal cord injury results in the loss of sensory,motor,and autonomic functions,which almost always produces permanent physical disability.Thus,in the search for more effective treatments than those already applied for years,which are not entirely efficient,researches have been able to demonstrate the potential of biological strategies using biomaterials to tissue manufacturing through bioengineering and stem cell therapy as a neuroregenerative approach,seeking to promote neuronal recovery after spinal cord injury.Each of these strategies has been developed and meticulously evaluated in several animal models with the aim of analyzing the potential of interventions for neuronal repair and,consequently,boosting functional recovery.Although the majority of experimental research has been conducted in rodents,there is increasing recognition of the importance,and need,of evaluating the safety and efficacy of these interventions in non-human primates before moving to clinical trials involving therapies potentially promising in humans.This article is a literature review from databases(PubMed,Science Direct,Elsevier,Scielo,Redalyc,Cochrane,and NCBI)from 10 years ago to date,using keywords(spinal cord injury,cell therapy,non-human primates,humans,and bioengineering in spinal cord injury).From 110 retrieved articles,after two selection rounds based on inclusion and exclusion criteria,21 articles were analyzed.Thus,this review arises from the need to recognize the experimental therapeutic advances applied in non-human primates and even humans,aimed at deepening these strategies and identifying the advantages and influence of the results on extrapolation for clinical applicability in humans.

Effects of P301L-TAU on post-translational modifications of microtubules in human iPSC-derived cortical neurons and TAU transgenic mice

TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal function and regulated via a complex set of post-translational modifications,changes of which affect microtubule stability and dynamics,microtubule interaction with other proteins and cellular structures,and mediate recruitment of microtubule-severing enzymes.As impairment of microtubule dynamics causes neuronal dysfunction,we hypothesize cognitive impairment in human disease to be impacted by impairment of microtubule dynamics.We therefore aimed to study the effects of a disease-causing mutation of TAU(P301L)on the levels and localization of microtubule post-translational modifications indicative of microtubule stability and dynamics,to assess whether P301L-TAU causes stability-changing modifications to microtubules.To investigate TAU localization,phosphorylation,and effects on tubulin post-translational modifications,we expressed wild-type or P301L-TAU in human MAPT-KO induced pluripotent stem cell-derived neurons(i Neurons)and studied TAU in neurons in the hippocampus of mice transgenic for human P301L-TAU(p R5 mice).Human neurons expressing the longest TAU isoform(2N4R)with the P301L mutation showed increased TAU phosphorylation at the AT8,but not the p-Ser-262 epitope,and increased polyglutamylation and acetylation of microtubules compared with endogenous TAU-expressing neurons.P301L-TAU showed pronounced somatodendritic presence,but also successful axonal enrichment and a similar axodendritic distribution comparable to exogenously expressed 2N4R-wildtype-TAU.P301L-TAU-expressing hippocampal neurons in transgenic mice showed prominent missorting and tauopathy-typical AT8-phosphorylation of TAU and increased polyglutamylation,but reduced acetylation,of microtubules compared with non-transgenic littermates.In sum,P301L-TAU results in changes in microtubule PTMs,suggestive of impairment of microtubule stability.This is accompanied by missorting and aggregation of TAU in mice but not in i Neurons.Microtubule PTMs/impairment may be of key importance in tauopathies.

Postnatal development of rat retina:a continuous observation and comparison between the organotypic retinal explant model and in vivo development

The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation.

The Human Sustainable Development under Biologically Evolutionary Background

Evolution,adaptation and selection always exist in the evolutionary process of any species,which is the result of interrelation and interaction between living things and their environments,and the incarnation of natural laws in the evolutionary process of any species. Only if accurately understanding their connotation and interrelation among them,taking the evolution of living things as the line with centralizing the human sustainable development,dialectically unifying the active adaptation of the human being to their environments and oriented selection of nature and the society to species,we can acquire a better understanding of evolution of living things and a deeper understanding of the human sustainable development under biologically evolutionary background and uphold the true Darwinism,modern evolutionary point of view and the concept of current sustainable development in theory and in practice.

Viewing Chinese modernization and Its Requirements for Human Rights Protection from the Perspective of Human Development

Modernization,as a specific stage in the historical development of human society,possesses a multi-layered composite structure,with each layer playing both a promoting and inhibiting role in human development and dignity simultaneously.Chinese modernization is a strategic choice made based on lessons learned from various experiences of modernization processes in other countries,reflecting judgments on the benefits and choices on the values of human development and dignity in modernization.It values the promotion of the free and well-rounded development of all individuals as its standard,demanding the full realization of the positive effects of modernization on human development and the overcoming of its negative effects.Human rights,aiming to promote the free and well-rounded development of every person as the ultimate value goal,are not only an intrinsic structural element of the modernization stage but also a social normative tool that restrains the direction of modernization development,aligning closely with the values pursued by Chinese modernization.Chinese modernization poses dual requirements for human rights protection:on the one hand,it demands the affirmation and promotion of the positive effects of modernization on human development and dignity through human rights standards,and on the other hand,it requires the prevention and mitigation of the negative effects of modernization on human development and dignity through human rights standards.There exists a certain tension between these two dimensions,necessitating an appropriate balance and thereby presenting corresponding requirements for China’s approach to human rights protection.

The relationship between DNA fragmentation and the intensity of morphologically abnormal human spermatozoa

Objective:To determine the relationship between teratozoospermia and sperm DNA fragmentation(SDF)in the human ejaculate.Methods:This retrospective study included 100 normozoospermic men as a control cohort(abnormal forms>14%),210 patients with a high level of abnormal forms(≤4%)and 65 patients presenting with a moderate level of abnormal forms(>4%to≤14%)based on the World Health Organization definitions.Sperm morphology was assessed using bright field microscopy.Sperm DNA fragmentation was assessed using the sperm chromatin dispersion assay.Non-parametric analyses were conducted to examine the relationship between abnormal sperm morphology and sperm DNA fragmentation;receiver operating characteristic(ROC)analyses were conducted to assess sensitivity and specificity of this relationship.Results:A correlation analysis revealed that the higher the proportion of abnormal spermatozoa in the ejaculate,the higher the level of SDF(Spearman's Rho=-0.230;P<0.001).Significant differences in the proportion of SDF were found when all cohorts were compared(P<0.001);these significant differences were also retained when the different cohorts were compared pairwise.ROC analysis showed a moderate but significant predictive value for SDF to differentiate patients with different levels of teratozoospemia.Conclusions:Although analysis of a more continuous range of values for teratozoospermia would help further clarify any causal relationship with SDF,there is clearly a synergistic or coincident affiliation between these variables that needs to be acknowledged by the clinician when interpreting the spermiogram.

Promote the All-round Development of China's Human Right Cause in the New Era——Summary of the Symposium on “Deeply Studying and Implementing the Spirit of the 20th CPC National Congress and Adhering to the Chinese Path of Human Rights Development”

On November 8, 2022, the symposium on “Deeply Studying and Implementing the Spirit of the 20thCPC National Congress and Adhering to the Chinese Path of Human Rights Development” was held by the China Society for Human Rights Studies in Beijing. Focusing on “the leadership of the CPC and human rights development”, “adapting Marxism to the Chinese context and the needs of our times and human rights theories in contemporary China”, “the Chinese path to modernization and the new form of human rights civilization”, “Chinese human rights solutions and global human rights governance”, “making a better China’s story of advancing human rights and constructing human rights discourse system”, and other topics, the participants reached broad consensus after discussion, and the symposium achieved fruitful results.

Changes in human settlement environments and their drivers in valley cities located in arid and semi-arid regions:A case study of Lanzhou in Western China

Development of urban human settlement environments(HSEs)is an integral part of promoting high-quality and sustainable regional development and constructing a beautiful China.The city of Lanzhou,located at the geometric center of China,is the only provincial capital traversed by the Yellow River.Given the constraints posed by the valley topography and the need for economic development,the development of this HSE,which is located within an arid region,poses considerable challenges.Evidently,an understanding of the evolution of HSEs and drivers of changes in them contributes to high-quality,sustainable urban development in arid and semi-arid regions.An analytical model was developed using the parameters of relief degree of land surface,human comfort days,the land cover index,nighttime light index,and precipitation.This model was used in combination with population density and the gross domestic product to analyze the spatial distribution of Lanzhou's HSE and its drivers.The results showed that landscapes in Lanzhou underwent significant changes between 2000 and 2022,with an increase in building-up land(+0.946%),cultivated land(+0.134%),and forest land(+0.018%)and a decrease in grassland(-1.10%).There was significant outward expansion of the main urban zone of Lanzhou and of various county towns,with the increase in building-up land being most prominent.During this period,there were significant changes in the periphery of the core urban area and county towns in Lanzhou,with decreases moving from the urban center(the highest value)to the surrounding areas(Yongdeng County had the lowest value).The correlation between the HSE and population density grew stronger in Anning and Chengguan Districts but became weaker in Xigu and Qilihe Districts.Spatiotemporal variations in the HSE were primarily caused by climate change,followed by human activities,and were also influenced by the valley topography.Overall,the spatial distribution of population density and the HSE in Lanzhou demonstrated good consistency under the in-fluence of economic development and urbanization.

Pioneering a New Realm of Human Rights Civilization in the Chinese Path to Modernization——An Overview of the Symposium on“The Chinese Path to Modernization and the Promotion of Free and Comprehensive Human Development”

On October 14,2023,a symposium themed“The Chinese Path to Modernization and the Promotion of Free and Comprehensive Human Development”was held in Changchun,Jilin Province,China.More than fifty experts,scholars,and researchers from national institutions and universities engaged in discussions and exchanges on human rights on the Chinese path to modernization,including the path,practice,knowledge systems,and civilization forms of human rights.This symposium played a significant role in advancing the construction of the disciplinary system,academic system,and discourse system of human rights in China.

Research on the Constitutional,Historical and Practical Logic of the Chinese Path of Human Rights Development——From the Perspective of the Inclusion of Human Rights Clause in the Constitution

The inclusion of the human rights clause in the Chi-nese Constitution is a concern and expression of the spirit of the Con-stitution,which has laid the institutional regulations of the fundamen-tal law for the development of human rights in China,provided the principles and value norms of the highest level of effectiveness for the legal protection of human rights,and built a profound constitutional basis for the formation and improvement of the Chinese path of human rights development.The human rights clause is not only a summariza-tion and affirmation of the historical practice of Chinese human rights development under the leadership of the Communist Party of China,but also a new starting point for the development of human rights in China under the leadership of the Communist Party of China.It marks that the development of human rights in China has entered a new era.The Party and the state have finally embarked on a path of human rights development with Chinese characteristics by formulating and implementing the Human Rights Action Plan of China,eliminating ab-solute poverty through the national poverty alleviation campaign,and promoting human rights protection through the rule of law.

Enhancing Erectile Function and Alleviating Andropause Symptoms: Clinical Efficacy of a Human Stem Cell Conditioned Medium Cream

Erectile dysfunction (ED) is increasingly prevalent in Japan, exceeding 30%, and increasing with age. Unhealthy lifestyle habits, obesity, insufficient exercise, and smoking have been implicated in its pathogenesis, along with endothelial dysfunction of the corpora cavernosa and impaired blood flow to the penis considered underlying factors. However, the current treatments are limited to Phosphodiesterase-5 (PDE5) inhibitors. ED is the primary symptom of andropathy. This study reports the clinical efficacy of human stem cell-conditioned medium cream for ED treatment. Ten men without underlying diseases suspected of andropause with ED (mean age 43.2 ± 4.4 y, Hb 15.2 ± 0.6 gm/dL, AST/ALT 30.2/37.9 ± 12.4/14.0, eGFR 82.7 ± 12.4 mL/min/1.73 m2) were targeted. The cream was applied twice daily to the genital and scrotal areas. The erectile hardness score (EHS), International Index of Erectile Function-5 (IIEF-5), and Aging Male Symptoms (AMS) scale were used to evaluate the participants before and 30 days after use, and the results were compared using paired t-tests. The post-use qualitative opinions were collected through interviews. Significant improvements were observed compared to baseline in the IIEF-5 (11.8 ± 4.6→17.2 ± 5.1, P < 0.001), and AMS (46.3 ± 6.7→37.6 ± 5.3, P < 0.001) scores post cream use. EHS did not show a statistically significant difference, but a trend towards improvement was observed. Qualitative feedback included increased morning erection, improved maintenance of erection during intercourse, and reduced post work fatigue. Human stem cell-conditioned medium contains endothelial growth factors that potentially contribute to the improvement of ED and andropause by enhancing corporal endothelial function. Future studies should include control groups to further investigate the efficacy of these treatments.

Impacts of Human Activities and Season on Species Diversity and Abundance of Butterflies in Mpanga Kipengere Game Reserve and Surrounding Farmlands, Tanzania

Maintaining natural habitats is crucial for the preservation of insects and other species that indicate environmental changes. However, the Mpanga/Kipengere Game Reserve and its surrounding farmlands are facing disturbance due to human activities, which is putting many wildlife species, particularly larger mammals, at risk. To determine the impact of human activities on butterfly species diversity and abundance in the reserve and its surrounding areas, we conducted a study from November 2021 to October 2023. We collected butterfly data using transect walks and baited traps in two habitat types. Our study yielded 2799 butterfly Individuals ranging in 124 species divided into five families habitat, season, and anthropogenic factors are significant environmental variables influencing species diversity and abundance of butterflies. Therefore, it’s important to protect habitat and dry-season water for the conservation of invertebrates such as butterflies. Our study findings provide essential information for ecological monitoring and future assessment of the Mpanga/Kipengere Game Reserve ecosystem health.