Extended protective effects of three dimensional cultured human mesenchymal stromal cells in a neuroinflammation model

BACKGROUND Human mesenchymal stromal cells(MSCs)possess regenerative potential due to pluripotency and paracrine functions.However,their stemness and immunomod-ulatory capabilities are sub-optimal in conventional two-dimensional(2D)culture.AIM To enhance the efficiency and therapeutic efficacy of MSCs,an in vivo-like 3D culture condition was applied.METHODS MSCs were cultured on polystyrene(2D)or in a gellan gum-based 3D system.In vitro,prostaglandin-endoperoxide synthase 2,indoleamine-2,3-dioxygenase,heme oxygenase 1,and prostaglandin E synthase gene expression was quantified by quantitative real-time polymerase chain reaction.MSCs were incubated with lipopolysaccharide(LPS)-treated mouse splenocytes,and prostaglandin E2 and tumor necrosis factor-alpha levels were measured by enzyme linked immuno-sorbent assay.In vivo,LPS was injected into the lateral ventricle of mouse brain,and MSCs were administered intravenously the next day.Animals were sacrificed and analyzed on days 2 and 6.RESULTS Gellan gum polymer-based 3D culture significantly increased expression of octamer-binding transcription factor 4 and Nanog homeobox stemness markers in human MSCs compared to 2D culture.This 3D environment also heightened expression of cyclooxygenase-2 and heme-oxygenase 1,enzymes known for immunomodulatory functions,including production of prostaglandins and heme degradation,respectively.MSCs in 3D culture secreted more prostaglandin E2 and effectively suppressed tumor necrosis factor-alpha release from LPS-stimulated splenocytes and surpassed the efficiency of MSCs cultured in 2D.In a murine neuroinflammation model,intravenous injection of 3D-cultured MSCs significantly reduced ionized calcium-binding adaptor molecule 1 and glial fibrillary acidic protein expression,mitigating chronic inflammation more effectively than 2D-cultured MSCs.CONCLUSION The microenvironment established in 3D culture serves as an in vivo mimetic,enhancing the immunomodulatory function of MSCs.This suggests that engineered MSCs hold significant promise a potent tool for cell therapy.

Current status and challenges for cell-cultured milk technology: a systematic review

Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture.While most cellular agriculture is predominantly centered on the production of cultured meat,there is a growing demand for an understanding of the production techniques involved in dairy products within cellular agriculture.This review focuses on the current status of cellular agriculture in the dairy sector and technical challenges for cell-cultured milk production.Cellular agriculture technology in the dairy sector has been classified into fermentation-based and animal cell culture-based cellular agriculture.Currently,various companies synthesize milk components through precision fermentation technology.Nevertheless,several startup companies are pursuing animal cell-based technology,driven by public concerns regarding genetically modified organisms in precision fermentation technology.Hence,this review offers an up-to-date exploration of animal cell-based cellular agriculture to produce milk components,specifically emphasizing the structural,functional,and productive aspects of mammary epithelial cells,providing new information for industry and academia.

Effect of Butternut Squash (Cucurbita moschata) Seed Powder on the Chemical and Rheological Properties of Stirred Cultured Camel Milk and Yoghurt

Research shows that producing fermented camel milk is hard because of the milk’s inability to form a firm coagulum, attributed to low levels of κ-casein and ꞵ-lactoglobulin and the large casein micelle size, leading to a weak network of casein formation. In an effort to address this issue, researchers turned to corn starch as a thickening agent, discovering that a concentration of 2.0% effectively improved the viscosity and significantly reduced syneresis in stirred camel milk yoghurt and cultured camel milk. This study explores alternatives to corn starch, focusing on butternut squash seeds as a promising substitute due to their hydrocolloid composition. By incorporating butternut squash (Cucurbita moschata) seed powder (BSSP) as a thickening agent, this study aimed at enhancing the chemical and rheological properties of stirred camel milk yoghurt and cultured camel milk. Fermented camel milk was prepared using 4 litres of camel milk, 2% starter cultures (thermophilic culture for yoghurt and mesophilic aromatic culture for stirred cultured camel milk) and BSSP 0.0% (negative control), 0.4%, 0.8%, 1.2%, 1.6%, 2.0% mixed with 0.4% gelatin. 2.0% corn starch mixed with 0.4% gelatin was used as a standard for comparison. Results showed that increasing the BSSP level significantly (p < 0.05) decreased the moisture content while increasing the total solid content of stirred fermented camel milk products. There was an increase in ash content with an increase in BSSP levels. There was a significant (p < 0.05) reduction in the pH, with an increase in BSSP levels in stirred fermented camel milk samples. Increasing the concentration of BSSP from 0.4% to 2.0% resulted in a significant (p < 0.05) increase in viscosity and a reduction in syneresis of stirred camel milk yoghurt and stirred cultured camel milk samples. This study demonstrated that BSSP effectively enhances the viscosity, reduces syneresis and increases acidity in stirred fermented camel milk products during storage.

Investigation of the Acceptability of Cultured Meat in University Students

Background: Over the past 20 years, cultured meat has drawn a lot of public attention as a potential solution to issues with animal husbandry, including inadequate use of natural sources, improper animal welfare practices, and possible risks to public health and safety. The novel method of producing meat through culture reduces the need for animals to produce muscle fiber, thereby obviating the necessity for animal slaughter. Apart from its ethical advantages, cultured meat presents a possible way to fulfill the expanding need for food among growing populations. The purpose of this research was to find out whether Turkish students would be willing to pay for and accept cultured meat. Technique: Method: 371 university students who willingly consented to fill out a questionnaire and provide demographic data make up the research sample. Questions from previous studies on the acceptability of cultured meat were compiled to create the survey. The research’s data collection took place in March and April of 2022. The research was completed in June 2022 after the data had been processed and analyzed. Results: The results showed that the majority of participants were female and had omnivorous eating habits. Based on the results of the Bonferroni correction test, students with a higher intention to purchase and consume cultured meat were those who received economics and business education. Students with two years of university education had a higher overall survey score than those with four years of education (p < 0.05). Furthermore, it is discovered that there is a negative correlation between the participants’ ages and their Factor 2 (using cultured meat as an alternative to industrial meat) and Factor 3 (consuming and purchasing it) section points (r = -109, p = 0.036) (r = -0.121, p = 0.019). In conclusion, university students generally have a negative outlook on health-related issues, such as eating cultured meat as an alternative.

“Culture time”板块教学中跨文化意识培养的策略

在新课程改革的大环境中,文化意识成为英语学科核心素养的重要组成部分。语言是文化的载体,语言与文化是相互依赖、相互影响、相互制约的。学习语言的过程也是学习文化的过程。文章主要结合译林版英语五、六年级教材中的Culture time板块的教学案例,探讨如何在英语课堂教学实践中丰富文化素材、发掘其中的文化元素和领略中西方文化异同,以帮助学生有效学习文化知识,拓宽文化视野,培养文化意识。

Effects of Estradiol on Cashmere Goat Primary Hair Follicles Cultured in vitro

[Objective] The aim of this study was to preliminarily explore the effects of estradiol on morphology and growth of cashmere goat primary hair follicles. [Method] Cashmere goat primary hair follicles were cultured in serum-free Williams E media supplemented with different doses of 17 β-E2 (0, 0.1, 1.0, 10.0, 100.0 nmol/L), and their growth rates and morphological changes were observed. [Result] The growth rate of 0.1 nmol/L 17 β-E2 group was quite comparable with that of the control group(0 nmol/L), but the 17 β-E2 with concentrations of 1.0, 10.0 and 100.0 nmol/L displayed different degrees of inhibition on the growth of hair follicles. Different morphological changes of hair follicles could also be discovered in different concentration treatments. [Conclusion] The study laid a certain foundation for exploring the regulation mechanism of estrogen on growth of cashmere goat hair follicles.

Propagation of Abies beshanzuensis by Water Cultured Medium

[Objective] The experiment aimed to explore the influences of phytohormones (ABT and IAA) and nutrient solution on rooting of Abies beshanzuensis M.H.Wu by water cultured medium. [Method] The Abies beshanzuensis M.H.Wu were treated by water (CK), 10 mg/L ABT+ water, 10 mg/L IAA+ water, 10 mg/L ABT+ hoagland solution, 10 mg/L IAA+ hoagland solution, then the rooting process was observed and the formation rate of callus, rooting rate, number of rooting, and root length were investigated and analyzed. [Result] ABT and IAA had obvious influences on callus induction, rooting rate and the number of root of Abies beshanzuensis M.H.Wu by water culture, so they were suitable to be used in water propagation of Abies beshanzuensis M.H.Wu. The treatments of phytohormones had no regular influences on the longest root length and average root length. The nutrient solutions would not generate obvious influence on propagation of Abies beshanzuensis M.H.Wu at firstly stage, but they generated influence on root growth after rooting. [Conclusion] The research provided new ideas for propagation of Abies beshanzuensis M.H.Wu, which could make it out of endangerment situation quickly.

Cultured meat from muscle stem cells: A review of challenges and prospects

Growing muscle tissue in culture from animal stem cells to produce meat theoretically eliminates the need to sacrifice animals. So-called ＂cultured＂ or ＂synthetic＂ or ＂in vitro＂ meat could in theory be constructed with different characteristics and be produced faster and more efficiently than traditional meat. The technique to generate cultured muscle tissues from stem cells was described long ago, but has not yet been developed for the commercial production of cultured meat products. The technology is at an early stage and prerequisites of implementation include a reasonably high level of consumer acceptance, and the development of commercially-viable means of large scale production. Recent advancements in tissue culture techniques suggest that production may be economically feasible, provided it has physical properties in terms of colour, flavour, aroma, texture and palatability that are comparable to conventional meat. Although considerable progress has been made during recent years, important issues remain to be resolved, including the characterization of social and ethical constraints, the fine-tuning of culture conditions, and the development of culture media that are cost-effective and free of animal products. Consumer acceptance and confidence in in vitro produced cultured meat might be a significant impediment that hinders the marketing process.

Experimental study of bioartificial liver with cultured human liver cells

AIM To establish an extracorporeal bioartificial liver support system (EBLSS) using cultured human liver cells and to study its support effect for fulminant hepatic failure (FHF).METHODS The liver support experiment of EBLSS consisting of aggregates cultured human liver cells, hollow fiber bioreactor, and circulation unit was carried out in dizhepatic dogs.RESULTS The viability of isolated hepatocytes and nonparenchymal liver cells reached 96%. These cells were successfully cultured as multicellular spheroids with synthetic technique. The typical morphological appearance was retained up to the end of the artificial liver experiment. Compared with the control dogs treated with EBLSS without liver cells, the survival time of artificial liver support dogs was significantly prolonged. The changes of blood pressure, heart rate and ECG were slow. Both serum ammonia and lactate levels were significantly lowered at the 3rd h and 5th h. In addition, a good viability of human liver cells was noted after 5 h experiment.CONCLUSION EBLSS playing a metabolic role of cultured human hepatocytes, is capable of compensating the function of the liver, and could provide effective artificial liver support and therapy for patients with FHF.

Challenges and prospects for consumer acceptance of cultured meat

Consumer acceptance of cultured meat is expected to depend on a wide diversity of determinants ranging from technologyrelated perceptions to product-specific expectations, and including wider contextual factors like media coverage, public involvement, and trust in science, policy and society. This paper discusses the case of cultured meat against this multitude of possible determinants shaping future consumer acceptance or rejection. The paper also presents insights from a primary exploratory study performed in April 2013 with consumers from Flanders（Belgium）（n=180）. The concept of cultured meat was only known（unaided） by 13% of the study participants. After receiving basic information about what cultured meat is, participants expressed favorable expectations about the concept. Only 9% rejected the idea of trying cultured meat, while two thirds hesitated and about quarter indicated to be willing to try it. The provision of additional information about the environmental benefits of cultured meat compared to traditional meat resulted in 43% of the participants indicating to be willing to try this novel food, while another 51% indicated to be ‘maybe＇ willing to do so. Price and sensory expectations emerged as major obstacles. Consumers eating mostly vegetarian meals were less convinced that cultured meat might be healthy, suggesting that vegetarians may not be the ideal primary target group for this novel meat substitute. Although exploratory rather than conclusive, the findings generally underscore doubts among consumers about trying this product when it would become available, and therefore also the challenge for cultured meat to mimic traditional meat in terms of sensory quality at an affordable price in order to become acceptable for future consumers.

Cloning of the non-structural gene 3 of hepatitis C virus and its inducible expression in cultured cells

AIM To study the inducible expression of hepatitis C virus ns3 gene (HCV ns3) in eukaryotic cells.METHODS The ns3 gene was obtained from plasmid pBns3 by polymerase chain reaction and inserted into the cloning vector pGEM-T. Then, the ns3 was subcloned into the vector pMSG to generate dexamethasone (DM)-inducible expression plasmid pMSG-ns3. CHO cells were transfected by pMSG-ns3 using calcium phosphate precipitation method and cultivated for 12 h-24 h. The transfected cells were induced with DM and the transient expression of NS3 protein was analyzed by ELISA and Western-blot methods.RESULTS After treated with 3×10-8mol/ L DM, the expression of NS3 was observed in the transfected CHO cells. A slightly higher level of NS3 was shown along with the time of DM treatment.CONCLUSION The inducible expressing vector pMSG-ns3 might be helpful for further studies of the characteristics of the ns3 gene in vivo.

Alternatives for large-scale production of cultured beef: A review

Cultured beef is a method where stem cells from skeletal muscle of cows are cultured in vitro to gain edible muscle tissue. For large-scale production of cultured beef, the culture technique needs to become more efficient than today＇s 2-dimensional（2D） standard technique that was used to make the first cultured hamburger. Options for efficient large-scale production of stem cells are to culture cells on microcarriers, either in suspension or in a packed bed bioreactor, or to culture aggregated cells in suspension. We discuss the pros and cons of these systems as well as the possibilities to use the systems for tissue culture. Either of the production systems needs to be optimized to achieve an efficient production of cultured beef. It is anticipated that the optimization of large-scale cell culture as performed for other stem cells can be translated into successful protocols for bovine satellite cells resulting in resource and cost efficient cultured beef.

Effect of endotoxin on fibronectin synthesis of rat primary cultured hepatocytes

AIM To investigate the effect of endotoxin on liver fibrosis and further define the role of hepatocytes in production of fibronectin in primary liver cell culture by endotoxin.METHODS After isolation and seeding of hepatocytes, the obtained cells were added to various doses (0, 5, 10, 15 and 20mg/L) of LPS treated culture media. The cells were collected and counted at various periods (0, 12, 24, 48, 72, 96, 120h). The concentrations of fibronectin were tested by electrophoresis.RESULTS The fibronectin levels tended to increase with prolongation of culture time. There was a sharp increase after 72h in 10 or 15 LPS treated group. The peak level of fibronectin was above 20mg/L. However, cell proliferation was inhibited during the course. Cell number of untreated control group (4.6±0.1×106) was about three-fold that of 20 LPS treated group (1.6±0.2×106) at 120h.CONCLUSION Hepatocytes have a potent ability to produce fibronectin stimulated by endotoxin, suggesting that hepatocytes might participate in the process of liver fibrosis.

Primary porcine hepatocytes with portal vein serum cultured on microcarriers or in spheroidal aggregates

AIM To develop a culture mode providingdurable biomaterials with high yields andactivities used in bioartificial liver.METHODS Hepatocytes were isolated from awhole pig liver by Seglen’s method of orthotopicperfusion with collagenase.In culture onmicrocarriers,primary porcine hepatocyteswere inoculated at a concentration of 5×10~7/mLinto the static culture systems containing 2 g/LCytodex-3,then supplemented with 100 mL/Lfetal calf serum(FCS)or 100 mL/L porcineportal vein serum(PPVS)respectively.Inspheroidal aggregate culture hepatocytes wereinoculated into 100 mL siliconized flasks at aconcentration of 5.0×10~6/mL.RESULTS In culture on microcarriershepatocytes tended to aggregate on Cytodex-3obviously after being inoculated.Typical multi-cellular aggregated spheroids could be found inthe two systems 24 h-48 h after hepatocyteswere cultured.The morphological charact-eristics and synthetic functions were maintainedfor 5 wk in FCS culture system and 8 wk in PPVSculture system.In spheroidal aggregate cultureabout 80%-90% isolated hepatocytes becameaggregated spheroids 24h after cultured insuspension and mean diameter of the spheroidswas 100μm.The relationship among thehepatocytes resembled that in the liver in vivo.Synthetic functions of albumin and urea of the spheroids were twice those of hepatocytescultured on monolayers.CONCLUSION As high-yields and high-activitymodes of culture on microcarriers or inspheroidal aggregate culture with portal veinserum are promising to provide biomaterials forbioartificial liver(BAL)efficiently.

Exosomes derived from 3D-cultured MSCs improve therapeutic effects in periodontitis and experimental colitis and restore the Th17 cell/Treg balance in inflamed periodontium

Although mesenchymal stem cell-derived exosomes(MSC-exos)have been shown to have therapeutic effects in experimental periodontitis,their drawbacks,such as low yield and limited efficacy,have hampered their clinical application.These drawbacks can be largely reduced by replacing the traditional 2D culture system with a 3D system.However,the potential function of MSC-exos produced by 3D culture(3D-exos)in periodontitis remains elusive.This study showed that compared with MSC-exos generated via 2D culture(2D-exos),3D-exos showed enhanced anti-inflammatory effects in a ligature-induced model of periodontitis by restoring the reactive T helper 17(Th17)cell/Treg balance in inflamed periodontal tissues.Mechanistically,3D-exos exhibited greater enrichment of miR-1246,which can suppress the expression of Nfat5,a key factor that mediates Th17 cell polarization in a sequence-dependent manner.Furthermore,we found that recovery of the Th17 cell/Treg balance in the inflamed periodontium by the local injection of 3D-exos attenuated experimental colitis.Our study not only showed that by restoring the Th17 cell/Treg balance through the miR-1246/Nfat5 axis,the 3D culture system improved the function of MSC-exos in the treatment of periodontitis,but also it provided a basis for treating inflammatory bowel disease(IBD)by restoring immune responses in the inflamed periodontium.

Structural Analysis of Water-soluble Polysaccharide PIP_1 Extracted from the Cultured Mycelium of Phellinus igniarius

Water-soluble crude polyseccharide（PIP） was extracted from cultured mycelium of the fungus Phellinus igniarius. After ethanol precipitation and sepharose CL-6B gel filtration, the fraction of PIP1 was obtained, which was shown to be a homogeneous polysaccharide by means of high-performance liquid chromatography. The structure of PIPt was determined by using several methods. C.,C analysis indicates that PIP1 is composed of the monosaccharides of glucose, galactose, and mannose. Their malar ratio is 3. 70： 4. 06： 1.00. The molar weight was estimated to be 17 kd via HPLC. IR, GC, partial hydrolysis with acid, pefiedate oxidation, Smith degradation, methylation, and GC-MS analysis were used for the structural analyses of PIP1. The results show that PIP1 has a small quantity of branch structure, The main glycosidic linkage of PIP1 has a β-configurafion. The main chain is made up of a large mass of glucose （ 1→3 ） and few mannose （ 1→4 ） ; the side chain is composed of glucose （ 1 →3 ） and galactose （ 1→6 ） ; the nonreduced end is composed of galactose and glucose. The side chains are branched at 6-0 of glucose（ 1→3,6） and mannose（1→4,6）. On an average, there are three branches among 20 residues. It is presumable that the existence of 1,3-linked Glc in the main and side chains is the main reason for its higher antitumor activity.

Effect of exogenous GA3 on flowering quality, endogenous hormones, and hormone-and flowering-associated gene expression in forcing-cultured tree peony（Paeonia suffruticosa）

Gibberellins(GAs)promote flowering in the forcing-cultured tree peony(Paeonia suffruticosa),however,the mechanism of regulating flowering is not fully understood.In this study,exogenous GA3 was applied to five-year-old Luoyang Hong plants to explore responses in terms of endogenous hormones,flowering quality,and the hormone-and flowering-associated gene expression.Exogenous GA3 application significantly promoted flower bud development and new branch growth,as well as improved flowering quality.Exogenous GA3 application also stimulated the synthesis of endogenous GA3 and indole-3-acetic acid(IAA)but reduced abscisic acid(ABA)levels.To further elucidate the regulatory mechanism,eight genes for GA biosynthesis and signaling,including PsCPS,PsKS,PsGA3ox,PsGA2ox,PsGID1b,PsGID1c,PsDELLA,and PsGID2 were cloned for the first time,and sequence analysis was also performed.The results suggested that all the cloned genes have conserved structure as each homologous gene reported in the other species.Phylogenetic trees constructed by the each cloned gene showed that the phylogenetic evolutionary relationship of P.suffruticosa was closely related to Vitis vinifera.The expression patterns of the above genes,and genes for ABA and IAA biosynthetic and signaling,and the flowering time were also investigated.Most of the above genes showed higher expression in the control buds than those in the GA3 treated buds at six developmental stages,whereas the expression levels of PsSOC1 and PsSPL9 were up-regulated by GA3 treatment.The results also showed that the GA-biosynthetic and signaling pathways are conserved in tree peony,and the PsCPS,PsGA3ox,PsGA2ox,PsGID1,PsDELLA,and PsGID2 genes are necessary for feedback regulation of GAs.Furthermore,hormone changes promoted PsSOC1 and PsSPL9 expression,and repressed PsSVP expression,which contributed to the improvement flowering quality in tree peony of forcing culture.

Effects of aminoguanidine on nitric oxide production induced by inflammatory cytokines and endotoxin in cultured rat hepatocytes

AIM: To study the effects of aminoguanidine (AG) and two L-arginine analogues N(omega)-nitro-L-arginine methyl ester (L-NAME) and N(omega)-nitro-L-arginine (L-NNA) on nitric oxide (NO) production induced by cytokines (TNF-alpha, IL-1 beta, and IFN-gamma) and bacterial lipopolysaccharide (LPS) mixture (CM) in the cultured rat hepatocytes, and examine their mechanisms action. METHODS: Rat hepatocytes were incubated with AG, L-NAME, L-NNA, Actinomycin D (ActD) and dexamethasone in a medium containing CM (LPS plus TNF-alpha, IL-1 beta, and IFN-gamma) for 24h. NO production in the cultured supernatant was measured with the Griess reaction. Intracellular cGMP level was detected with radioimmunoassy. RESULTS: NO production was markedly blocked by AG and L-NAME in a dose-dependent manner under inflammatory stimuli condition triggered by CM in vitro. The rate of the maximum inhibitory effects of L-NAME (38.9%) was less potent than that obtained with AG(53.7%, P 【 0.05). There was no significant difference between the inhibitory effects of AG and two L-arginine analogues on intracellular cGMP accumulation in rat cultured hepatocytes. Non-specific NOS expression inhibitor dexamethasone (DEX)and iNOS mRNA transcriptional inhibitor ActD also significantly inhibited CM-induced NO production. AG(0.1 mmol x L(-1)) and ActD (0.2 ng x L(-1)) were equipotent in decreasing NO production induced by inflammatory stimuli in vitro, and both effects were more potent than that induced by non-selectivity NOS activity inhibitor L-NAME (0.1 mmol x L(-1)) under similar stimuli conditions (P【0.01). CONCLUSION: AG is a potent selective inhibitor of inducible isoform of NOS,and the mechanism of action may be not only competitive inhibition in the substrate level, but also the gene expression level in rat hepatocytes.

The environmental prospects of cultured meat in China

To deal with concerns in China about environmental degradation and a growth in population accompanied by increased consumption of livestock products, a meat alternative is required. This study compared the environmental impacts of producing different protein sources for nutrition, including crops, livestock products, and cultured meat. The results showed that cultured meat has the lowest land use per unit of protein and unit of human digestible energy. China＇s crops have the lowest energy use and greenhouse gas（GHG） emissions per unit of energy and protein. The energy use in cultured meat production is slightly higher than that of current pork production in China, whereas GHG emissions are lower. It is concluded that the overall impact of replacing livestock products with cultured meat would be beneficial for China＇s environment and would potentially improve food security because less land is needed to produce the same amount of protein and energy.

Oxidative damage of primary cultured hippocampal neurons Does androgen have an antagonistic effect?

BACKGROUND： Evidence illustrates that androgen has a neuroprotective role. However, whether androgen also has the protective effect on hippocampal neurons during free radical mediated injury remains unclear. OBJECTIVE： To investigate the neuroprotective effect of androgen on hippocampal neurons during free radical damage. DESIGN, TIME AND SETTING： A controlled in vitro experiment was performed at the Department of Human Anatomy, Cell Culture Lab, and Neuroendocrinology Lab, Basic Medical School, Hebei Medical University from February to June 2009. MATERIALS： Testosterone was provided by Tianjin Jinyao Amino Acid Company, China. METHODS： Primary cultured neurons from 24 Sprague Dawley rats were randomly assigned into four groups： control, H202, testosterone, and testosterone （pre-added） plus H2O2 groups. MAIN OUTCOME MEASURES： The positive cell ratio of microtubule associated protein-Ⅱ and neuron specific enolase was determined by immunocytochemistry. Neuronal morphology was observed by hematoxylin-eosin staining and Nissl staining. Cell vitality and viability were determined using an inverted phase contrast microscope. The content of nitric oxide synthase, malondialdehyde, and superoxide dismutase were measured with a spectrophotometer. RESULTS： As compared with the control group, cell vitality and viability, and superoxide dismutase level were significantly decreased in the H202 group （P 〈 0.05）, while nitric oxide synthase and malondialdehyde levels were significantly increased （P 〈 0.05）. Neuronal vitality and viability as well as superoxide dismutase level in the testosterone plus H2O2 group were significantly greater than in the H2O2 group （P 〈 0.05）, and nitric oxide synthase and malondialdehyde levels were significantly less than in the H2O2 group （P〈 0.05）. CONCLUSION： Androgen partially reversed H2O2-induced neuronal damage and protected neurons.