Nogo-A Drives Alzheimer's Disease Progression by Inducing Tauopathy Vulnerability.

Tauopathies, a group of neurodegenerative disorders, are characterized by disrupted homeostasis of the microtubule binding protein tau. Nogo-A mainly hinders axonal growth and development in neurons, but the underlying mechanism of tau vulnerability has not been determined. Here, to gain more comprehensive insights into the impact of Nogo-A on tau protein expression, we showed that Nogo-A induces tau hyperphosphorylation, synapse loss and cognitive dysfunction. Consistent with the biological function of tau hyperphosphorylation, Nogo-A-induced tau hyperphosphorylation altered microtubule stability, which causes synaptic dysfunction. Mechanistically, Nogo-A-induced tau hyperphosphorylation was abolished by the Nogo-A antagonist NEP1-40 in primary neurons. Surprisingly, downregulation of Nogo-A in the hippocampus of AD mice (hTau. P301S) inhibited tau hyperphosphorylation at the AT8, Thr181, The231 and Ser404 sites and rescued synaptic loss and cognitive impairment in AD mice. Our findings exhibit a strong degree of consistency with Nogo-A-induced tauopathy vulnerability, reinforcing the coherence and reliability of our research. Furthermore, in mice, Nogo-A increases tauopathy vulnerability to exacerbate AD progression via ROCK/AKT/GSK3ß signaling. Together, our findings provide new insight into the function of Nogo-A in regulating tau hyperphosphorylation and reveal an effective treatment strategy for tauopathies.

EGFR-Activated JAK2/STAT3 Pathway Confers Neuroprotection in Spinal Cord Ischemia-Reperfusion Injury: Evidence from High-Throughput Sequencing and Experimental Models.

This study aimed to investigate the molecular mechanisms underlying spinal cord ischemia-reperfusion (SCI/R) injury. Through RNA-Seq high-throughput sequencing and bioinformatics analysis, we found that EGFR was downregulated in the spinal cord of SCI/R mice and may function via mediating the JAK2/STAT3 signaling pathway. In vitro cell experiments indicated that overexpression of EGFR activated the JAK2/STAT3 signaling pathway and reduced neuronal apoptosis levels. In vivo animal experiments further confirmed this conclusion, suggesting that EGFR inhibits SCI/R-induced neuronal apoptosis by activating the JAK2/STAT3 signaling pathway, thereby improving SCI/R-induced spinal cord injury in mice. This study revealed the molecular mechanisms of SCI/R injury and provided new therapeutic strategies for treating neuronal apoptosis.

Effects of different grassland utilization methods on the germinable soil seed bank of the Hulunbuir meadow steppe.

Seed banks are crucial regenerative resources for aboveground vegetation. The pattern of their changes holds immense significance in understanding alterations in the belowground seed bank. This understanding is pivotal for uncovering both short-term and long-term shifts in plant communities. Additionally, it contributes to the restoration of grassland ecosystems. To better protect grassland biodiversity and provide a theoretical basis for the restoration of degraded grasslands, in this study, the germination characteristics of soil seed banks in free-grazed, enclosed and mown areas were compared, and the results were combined with those of previous studies for a comprehensive analysis. The density of soil seed bank and perennial forage soil seed bank were significantly affected by different grassland utilization and soil depths. Grazing and enclosure grassland utilization methods increased the content of the soil seed bank, and mowing reduced the content of the seed bank. The soil seed bank density of perennial grasses accounted for the highest proportion under grazing, followed by mowing, and its lowest proportion was observed in the enclosures. Grazing not only facilitated the germination of the perennial grass seed bank but also substantially augmented its content. Mowing inhibited the germination of the upper growth grasses seed bank, which was particularly significant in the 0-2 cm soil layer under grazing. The content of the upper growth grasses seed bank affected the total seed bank to a certain extent, mainly in the 5-10 cm layer. The general correlations among the perennial grasses, upper growth grasses and soil germination seed bank resulted in 84.58% information extraction, and this information has practical significance for grassland ecological restoration.

Heavy grazing causes plant cluster fragmentation of sparse grasses.

Cleistogenes songorica, as a clustered grass, is the main grassland flora of the Stipa breviflora desert grassland. Some studies have shown that the constructive species of S. breviflora (sparse cluster type) is prone to cluster fragmentation; however, research on C. songorica is relatively rare. Then will the C. songorica plant population (dense cluster type) also have cluster fragmentation under the influence of intense grazing? To answer this question, we used variance analysis and geo-statistical methods. The spatial distribution of C. songorica in S. breviflora desert steppe in Inner Mongolia was measured under four grazing intensities (no grazing, CK, 0 sheep·ha-1·half year-1; light grazing, LG, 0.93 sheep·ha-1·half year-1; moderate grazing, MG, 1.82 sheep·ha-1·half year-1; and heavy grazing, HG, 2.71 sheep·ha-1·half year-1) and four scales (10 cm × 10 cm, 20 cm × 20 cm, 25 cm × 25 cm, 50 cm × 50 cm). We then analyzed C. songorica whether fragmentation was present. The results showed that increased grazing intensity is associated with increased density and decreased height, coverage, and standing crop of C. songorica. The spatial distribution of C. songorica was affected by structural factors, and spatial heterogeneity decreased with increased spatial scale. With increased grazing intensity and spatial scale, the patch area of C. songorica gradually increased and tended toward band distribution. In summary, increased grazing intensity and spatial scale led to weakened heterogeneity of C. songorica spatial distribution and increased consistency.

Immune mechanism of low bone mineral density caused by ankylosing spondylitis based on bioinformatics and machine learning.

Background and Objective: This study aims to find the key immune genes and mechanisms of low bone mineral density (LBMD) in ankylosing spondylitis (AS) patients. Methods: AS and LBMD datasets were downloaded from the GEO database, and differential expression gene analysis was performed to obtain DEGs. Immune-related genes (IRGs) were obtained from ImmPort. Overlapping DEGs and IRGs got I-DEGs. Pearson coefficients were used to calculate DEGs and IRGs correlations in the AS and LBMD datasets. Louvain community discovery was used to cluster the co-expression network to get gene modules. The module most related to the immune module was defined as the key module. Metascape was used for enrichment analysis of key modules. Further, I-DEGs with the same trend in AS and LBMD were considered key I-DEGs. Multiple machine learning methods were used to construct diagnostic models based on key I-DEGs. IID database was used to find the context of I-DEGs, especially in the skeletal system. Gene-biological process and gene-pathway networks were constructed based on key I-DEGs. In addition, immune infiltration was analyzed on the AS dataset using the CIBERSORT algorithm. Results: A total of 19 genes were identified I-DEGs, of which IFNAR1, PIK3CG, PTGER2, TNF, and CCL3 were considered the key I-DEGs. These key I-DEGs had a good relationship with the hub genes of key modules. Multiple machine learning showed that key I-DEGs, as a signature, had an excellent diagnostic performance in both AS and LBMD, and the SVM model had the highest AUC value. Key I-DEGs were closely linked through bridge genes, especially in the skeletal system. Pathway analysis showed that PIK3CG, IFNAR1, CCL3, and TNF participated in NETs formation through pathways such as the MAPK signaling pathway. Immune infiltration analysis showed neutrophils had the most significant differences between case and control groups and a good correlation with key I-DEG. Conclusion: The key I-DEGs, TNF, CCL3, PIK3CG, PTGER2, and IFNAR1, can be utilized as biomarkers to determine the risk of LBMD in AS patients. They may affect neutrophil infiltration and NETs formation to influence the bone remodeling process in AS.

Fluoxetine modifies circadian rhythm by reducing melatonin content in zebrafish.

Fluoxetine (FLX), a selective serotonin reuptake inhibitor (SSRI), increases the serotonin levels in the brain to treat depression. Antidepressants have been demonstrated to modulate circadian rhythm, but the underlying mechanisms by which antidepressants regulate circadian rhythm require more research. This study aimed to investigate the role of FLX on circadian rhythm by analyzing the movement behavior and internal circadian oscillations in zebrafish. The results showed that the expression of clock genes clock1a and bmal1b was significantly down-regulated, and the amplitude reduction and phase shift were observed after FLX treatment. Furthermore, FLX exposure inhibited the expression of aanat2, which led to a decrease in nocturnal melatonin secretion. aanat2-/- larvae showed disrupted circadian rhythm. These findings may help reveal the effect of FLX exposure on the circadian rhythm and locomotor activity. It may provide theoretical data for the clinical application of FLX.

Heavy grazing reduced the spatial heterogeneity of Artemisia frigida in desert steppe.

BACKGROUND: Grazing disturbance plays an important role in the desert steppe ecosystem in Inner Mongolia, China. Previous studies found that grazing affected the spatial distribution of species in a community, and showed patchiness characteristics of species under different grazing treatments. Artemisia frigida is the dominant species and semi-shrub in desert steppe, and whether grazing interference will affect the spatial distribution of A. frigida is studied. In this study, geo-statistical methods were mainly used to study the spatial distribution characteristics of A. frigida population in desert steppe of Inner Mongolia at two scales (quadrat size 2.5 m × 2.5 m, 5 m × 5 m) and four stocking rates (control, CK, 0 sheep·ha-1·month-1; light grazing, LG, 0.15 sheep·ha-1·month-1, moderate grazing, MG, 0.30 sheep·ha-1·month-1, heavy grazing, HG, 0.45 sheep·ha-1·month-1). RESULTS: The results showed that the spatial distribution of A. frigida tended to be simplified with the increase of stocking rate, and tended to be banded with increased spatial scale. The density and height of A. frigida increased with increasing scale. With increased stocking rate, the density of A. frigida population decreased linearly, while its height decreased in a step-wise fashion. The spatial distribution of A. frigida was mainly affected by structural factors at different scales and stocking rate. The density of A. frigida was more sensitive to change in stocking rate, and the patchiness distribution of A. frigida was more obvious with increase in scale. CONCLUSIONS: Stocking rate has a strong regulatory effect on the spatial pattern of A. frigida population in the desert steppe. Heavy grazing reduced the spatial heterogeneity of A. frigida in the desert steppe. The smaller dominant populations are unfavourable for its survival in heavy grazing condition, and affects the stability and productivity of the grassland ecosystem.

Muscarinic acetylcholine receptors regulate inflammatory responses through arginases 1/2 in zebrafish.

Muscarinic acetylcholine receptors (mAChRs) are widely expressed in various effector cells and have been proved to play vital roles in smooth muscle contraction and digestive secretion. However, there are relatively few literatures revealing the roles of mAChRs in inflammatory processes, and its underlying regulatory mechanisms have not been elucidated. Taking the advantages of live imaging of zebrafish, we found that inhibition of mAChRs resulted in increased neutrophils recruitment and proinflammatory cytokines expression, whereas activation of mAChRs led to opposite outcome. Subsequently, we found that mAChRs regulated the expression of arginases (args), and pharmacological intervention of args level could reverse the effects of mAChRs on neutrophils migration and cytokines expression, suggesting that args are important downstream proteins of mAChRs that mediate the regulation of inflammatory response. In this study, we identified args as novel downstream proteins of mAChRs in inflammatory responses, providing additional evidence for system immune regulation of cholinergic receptors.

Hypoxia regulates cytokines expression and neutrophils migration by ERK signaling in zebrafish.

Normal dissolved oxygen in water is essential for maintaining the physiological functions of fish, but environmental pollution, such as eutrophication can lead to a decrease in oxygen content in water. How this reduction of dissolved oxygen in water affects the immune functions of fish and the potential regulatory mechanisms have not been thoroughly elucidated. In this study, we made full use of the aquatic model animal zebrafish to explore this question. In a model of LPS-induced inflammation, we found that hypoxia induced by infusing nitrogen into water increased the expression of pro-inflammatory cytokines, such as il-1ß, il-6, and il-8. In vivo imaging also showed that hypoxia significantly increased neutrophil migration to the site of caudal fin injury in the transgenic line. Subsequently, we found that the phosphorylation level of ERK protein was significantly activated upon hypoxia and proved the roles of ERK signaling in the expression of pro-inflammatory cytokines and neutrophil migration in zebrafish. This study indicated that reduced water oxygen significantly increases the inflammatory response of the zebrafish.

Study on the stability and cellular affinity of gelatin-polysaccharide composite films.

The gelatin film has great potential in biomedical applications, especially in wound healing. The combination of gelatin films and stem cells could further accelerate the skin regeneration. Although polysaccharide modification can improve the mechanical property and biological activity of gelatin films, information about the stability and cellular affinity is still limited. This study investigated the influence of polysaccharides on the stability and cellular affinity of gelatin films. Two kinds of gelatin-polysaccharide composite films, including gelatin-hyaluronic acid (G-HA) and gelatin-chitosan (G-CS), were prepared in this study. It was found that G-HA composite film had better short-term and long-term stability compared with G-CS composite film. And G-HA composite film also had better biological safety than G-CS film. Moreover, the surface of G-HA composite film supported the adhesion and growth of human umbilical cord Wharton's jelly-derived mesenchymal stem cells (WJ MSCs) better than G-CS film surface. These data illustrated that G-HA composite film has better stability and cellular affinity compared with G-CS film, which could be considered a promising delivery system of stem cells for further in vivo studies. Therefore, this work would be very helpful to optimize the preparation of gelatin-polysaccharide composite films.

Identification of foam cell biomarkers by microarray analysis.

BACKGROUND: Lipid infiltration and inflammatory response run through the occurrence of atherosclerosis. Differentiation into macrophages and foam cell formation are the key steps of AS. Aim of this study was that the differential gene expression between foam cells and macrophages was analyzed to search the key links of foam cell generation, so as to explore the pathogenesis of atherosclerosis and provide targets for the early screening and prevention of coronary artery disease (CAD). METHODS: The gene expression profiles of GSE9874 were downloaded from Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE9874) on GPL96 [HG-U133A] Affymetrix Human Genome U133. A total of 22,383 genes were analyzed for differentially expression genes (DEGs) by Bayes package. GO enrichment analysis and KEGG pathway analysis for DEGs were performed using KOBAS 3.0 software (Peking University, Beijing, China). STRING software (STRING 10.0; European Molecular Biology Laboratory, Heidelberg, Germany) was used to analyze the protein-protein interaction (PPI) of DEGs. RESULTS: A total of 167 DEGs between macrophages and foam cells were identified. Compared with macrophages, 102 genes were significantly upregulated and 65 genes were significantly downregulated (P < 0.01, fold-change > 1) in foam cells. DEGs were mainly enrich in 'sterol biosynthetic and metabolic process', 'cholesterol metabolic and biosynthetic process' by GO enrichment analysis. The results of KEGG pathway analysis showed all differential genes are involved in biological processes through 143 KEGG pathways. A PPI network of the DEGs was constructed and 10 outstanding genes of the PPI network was identified by using Cytoscape, which include HMGCR, SREBF2, LDLR, HMGCS1, FDFT1, LPL, DHCR24, SQLE, ABCA1 and FDPS. CONCLUSION: Lipid metabolism related genes and molecular pathways were the key to the transformation of macrophages into foam cells. Therefore, lipid metabolism disorder is the key to turn macrophages into foam cells, which plays a major role in CAD.

Dominant species' dominant role and spatial stability are enhanced with increasing stocking rate.

Stipa breviflora Grisb. (S. breviflora) is a dominant species in the desert steppe of northern China. Its function and role at the plant community level increases with increasing stocking rate. However, the response of spatial stability remains unclear. We selected treatment areas representing no grazing (CK), light grazing (LG), moderate grazing (MG) and heavy grazing (HG) in a long-term grazing experiment (2004-2017) in a S. breviflora desert steppe in Inner Mongolia, northern China. Using a mechanical sampling method, 40 m × 40 m representative sample plots were selected to obtain the height, coverage and density of the S. breviflora population and community, and we computed the standing crop of mechanical sampling quadrats based on a random sample of cutting quadrats. Analysis of standing crop, density of S. breviflora population and its ratio in the plant community showed that the dominant role of S. breviflora population in the plant community increased with increasing grazing intensity, while the spatial stability of S. breviflora population not only had many dimensions, but also many states. The dimension or combination of dimensions of its stability performance and its adaptive state varied under different disturbance intensities and frequencies.

Congenital unilateral proximal radioulnar synostosis: A surgical case report.

RATIONALE: Congenital proximal radioulnar synostosis is a rare genetic malformation of the upper limb. This deformity, which is found mainly in preschool-aged children, has no recognized diagnosis and treatment. Current diagnostic methods cannot effectively assess both bone structure and soft tissue abnormalities, and most surgical treatments introduce complications and do not prevent recurrence. More work is needed; therefore, to address the diagnosis and treatment of this disease. PATIENT CONCERNS: An 8-year-old male patient was hospitalized in our department. He reported deformity and limited motion in his right elbow for the past 2 years. He denied a traumatic or family history of bony malformation. The chief complaint at the time of the hospitalization was the limitation in forearm rotation. DIAGNOSIS: Digital radiography of the right elbow joint showed proximal radioulnar synostosis and a valgus deformity. A 3-dimensional computed tomography scan further showed proximal ulna and radius dysplasia as well as anterior dislocation of the radius head. The patient was diagnosed with congenital right proximal radioulnar synostosis. INTERVENTIONS: Surgical procedures included arthrolysis of the right proximal radioulnar joint, osteotomy of the proximal radius, internal fixation with Kirschner wires, and reconstruction of the annular ligament. The right elbow was immobilized in plaster in a flexion and supination position for 2 weeks. OUTCOMES: Recurrence of the right proximal radioulnar synostosis was observed during the 6-month follow-up, but the rotation function of the patient's forearm was significantly improved. LESSONS: The findings from this case suggest that we should carefully monitor all patients younger than 6 years old who report long-term issues with forearm rotation. This case also highlights the need to assess soft tissue and epiphysis abnormalities in addition to bone assessments via digital radiography and 3-dimensional computed tomography. We suggest that surgery should not be performed until the proximal radius epiphysis has closed. Not all cases require surgical treatment, but when surgery is needed, a suitable method should be selected according to the individual needs of the patient. Any surgery performed should treat both the bony malformations and soft tissue abnormalities to maximize the therapeutic effect and reduce complications during and after surgery.

Differential distributions of parvalbumin-positive interneurons in the sulci and gyri of the adult ferret cerebral cortex.

Although accumulating evidence suggests that there are significant anatomical and histological differences between the sulci and gyri of the cerebral cortex, whether there is a difference in the distribution of interneurons between the two cortical regions remains largely unknown. In this study, we systematically compared the distributions of parvalbumin-positive interneurons among three neighboring gyrus and sulcus pairs-coronal gyrus and cruciate sulcus, anterior ectosylvian gyrus and rostral suprasylvian sulcus, and posterior ectosylvian gyrus and pseudosylvian sulcus-in the adult ferret cerebral cortex. We proposed a method to partition sulci and gyri into several specific subregions through the deepest points of the sulci and the highest points of gyri in the inner and outer cortical contours of coronal sections. We found that the density of parvalbumin-positive interneurons in the gyri was significantly higher than that in the sulci. Further study revealed that the density of PV interneurons in superficial cortical layers (layers 2/3 and layer 4) was comparable among the three pairs of sulci and gyri. However, the density of parvalbumin-positive interneurons in cortical layers 5/6 was significantly higher in gyri than in sulci. These results indicate that parvalbumin-positive interneurons are differently distributed in infragranular layers of cortical sulci and gyri.

Grazing intensity enhances spatial aggregation of dominant species in a desert steppe.

Understanding how grazing activity drives plant community structure or the distribution of specific species in a community remains a major challenge in community ecology. The patchiness or spatial aggregation of specific species can be quantified by analyzing their relative coordinates in the community. Using variance and geostatistical analysis methods, we examined the quantitative characteristics and spatial distribution of Stipa breviflora in a desert steppe in northern China under four different grazing intensities (no grazing, NG, light grazing, LG, moderate grazing, MG, and heavy grazing, HG) at three small spatial scales (10 × 10 cm, 20 × 20 cm, 25 × 25 cm). We found that grazing significantly increased cover, density, and proportion in standing crop of S. breviflora, but decreased height. The spatial distribution of S. breviflora was strongly dependent upon the sampling unit and grazing intensity. The patchiness of S. breviflora reduced with sampling scale, and spatial distribution of S. breviflora was mainly determined by structural factors. The intact clusters of S. breviflora were more fragmented with increasing grazing intensity and offspring clusters spread out from the center of the parent plant. These findings suggest that spatial aggregation can enhance the ability of S. breviflora to tolerate grazing and that smaller isolated clusters are beneficial to the survival of this dominant species under heavy grazing.

Syncope as the initial presentation of pulmonary embolism in a young adult with testicular tumor: A case report and literature review.

RATIONALE: Venous thrombus embolism (VTE) includes deep-vein thrombosis (DVT) and pulmonary embolism (PE) which may be an initial symptom for patients with cancer. PE has diverse clinical manifestations and is a rare complication of testicular tumor (TT). PATIENT CONCERNS: Here, we report a 21-year-old man admitted to our hospital due to syncope. DIAGNOSES: Clinical examinations upon admission demonstrated PE resulting in syncope. Further, a malignant TT, liver metastasis, and inferior vena cava (IVC) thrombosis were diagnosed. INTERVENTIONS: Low molecular heparin was administered immediately after PE was diagnosed. OUTCOMES: The patient suffered from cardiac arrest on hospitalization. LESSONS: Physicians should consider the possibility of TT when a young male patient presents with syncope and is diagnosed with PE that cannot be explained by a common cause. Treatment for TT and PE should be performed as early as possible to improve the prognosis of patients combine with TT and PE.

Managed grassland alters soil N dynamics and N2O emissions in temperate steppe.

Reclamation of degraded grasslands as managed grasslands has been increasingly accelerated in recent years in China. Land use change affects soil nitrogen (N) dynamics and nitrous oxide (N2O) emissions. However, it remains unclear how large-scale grassland reclamation will impact the grassland ecosystem as a whole. Here, we investigated the effects of the conversion from native to managed grasslands on soil N dynamics and N2O emissions by field experiments in Hulunber in northern China. Soil (0-10cm), nitrate (NO3-), ammonium (NH4+), and microbial N were measured in plots in a temperate steppe (Leymus chinensis grassland) and two managed grasslands (Medicago sativa and Bromus inermis grasslands) in 2011 and 2012. The results showed conversion of L. chinensis grassland to M. sativa or B. inermis grasslands decreased concentrations of NO3--N, but did not change NH4+-N. Soil microbial N was slightly decreased by the conversion of L. chinensis grassland to M. sativa, but increased by the conversion to B. inermis. The conversion of L. chinensis grassland to M. sativa (i.e., a legume grass) increased N2O emissions by 26.2%, while the conversion to the B. inermis (i.e., a non-legume grass) reduced N2O emissions by 33.1%. The conversion from native to managed grasslands caused large created variations in soil NO3--N and NH4+-N concentrations. Net N mineralization rates did not change significantly in growing season or vegetation type, but to net nitrification rate. These results provide evidence on how reclamation may impact the grassland ecosystem in terms of N dynamics and N2O emissions.

Dihydroartemisinin induces apoptosis and downregulates glucose metabolism in JF-305 pancreatic cancer cells.

Cancer cell promotion of glycolysis provides a promising therapeutic target for cancer treatment. Dihydroartemisinin (DHA) displays cytotoxicity to multiple human tumor cells. However, its effects on pancreatic cancer cells are not well studied. The objective of this study was to investigate the effect of DHA on glucose metabolism and cell viability in JF-305 pancreatic cancer cells. To achieve these goals, cell viability was measured with MTT assay, and the occurrence of apoptosis was detected. Glucose uptake, lactate production, and ATP content were measured. Western blotting was used for the detection of apoptosis-related protein expression. The result showed that DHA caused significant reduction in JF-305 cell viability, arrested the cell phase in G2/M, induced apoptosis, and decreased the mitochondrial membrane potential and accumulated ROS. DHA also inhibited glucose uptake, lactate generation, and ATP production. Western blotting showed that treatment with DHA increased the activity of caspase-9 and caspase-3, downregulated Bcl-2 expression, and upregulated the expression levels of Bax and Cyto C. Meanwhile, DHA downregulated the Akt/mTOR signaling pathway and inhibited glucose transporter 1 expression. Our data suggest that DHA treatment increased the apoptosis of JF-305 pancreatic cancer cells, and the effect of apoptosis may be associated with the inhibition of glycolysis.

[Dihydroartemisinin inhibits proliferation of pancreatic cancer JF-305 cells by regulating expression of apoptosis related proteins and production of reactive oxygen species].

To investigate the effect of dihydroartemisinin on apoptosis of human pancreatic cancer cell line JF-305 and the role of reactive oxygen species(ROS) in the apoptosis of JF-305 cells induced by dihydroartemisinin. MTT assays were used to detect effect of different concentrations of dihydroartemisinin on cells proliferation of JF-305 lines. Cell cycle was detected by flow cytometry, and the apoptotic morphology was observed by Hoechst 333258 fluorescence staining. Annexin V fluorescence staining was used to detect the apoptosis changes of JF-305 cells, while DCFH-DA was used to detect the changes of ROS during apoptosis process. Western blot was used to detect the protein expression changes of Bax, Bcl-2, Cleaved caspase-3, Cleaved caspase-9 and Cyto C. As compared with the control group, the JF-305 cells proliferation was inhibited significantly(P<0.05) after treatment with different concentrations of dihydroartemisimin for 48 h; cell cycle was blocked in the G2/M phase; apoptotic morphology of nuclear condensation, aggregation, and fragmentation was found, and the apoptosis ratio was increased(P<0.05). DCFH-DA detection showed that the cell ROS was increased significantly after dihydroartemisinin treatment(P<0.05). Western blot results showed that the expression of Bcl-2 protein was down-regulated; the expression of Bax protein was up-regulated; the ration of Bax/Bcl-2 was increased and the protein expression levels of Cleaved caspase-3, Cleaved caspase-9 and Cyto C were increased after dihydroartemisinin treatment. Therefore, dihydroartemisinin could induce apoptosis of JF-305 cells, and the possible mechanism may be related to the formation and increasing of ROS.

Effects of Saikosaponin D on apoptosis in human U87 glioblastoma cells.

Saikosaponin D (SSd) is a type of Saponin derivative, which is a component extracted from Bupleurum falactum. SSd has been reported to exert anticancer activities. However, the effects of SSd on gliomas have not been elucidated. The aim of the present study was to investigate the pharmacological functions and potential molecular mechanisms of SSd in human U87 glioblastoma cells. The cells were treated with SSd at various concentrations for 48 h, the cell viability was assessed with 3­(4,5­dimethylthiazol­2­yl)­2,5­diphenyltetrazolium assay, and the activation of Akt, extracellular signal­regulated kinases (ERK), c­Jun N­terminal kinases (JNK) and caspase­3 was assessed by western blotting. In addition, apoptosis levels were analyzed with Hoechst 33258 and Annexin V staining. The results demonstrated that treatment of the U87 glioma cells with SSd markedly suppressed cell proliferation in a dose­dependent manner. Meanwhile, SSd treatment enhanced apoptosis in the U87 cells. Furthermore, SSd significantly inhibited the phosphorylation of Akt and ERK, and promoted phosphorylated­JNK and cleaved caspase­3 expression. The present study revealed the potential therapeutic effects of SSd in the treatment of gliomas, and the cytotoxic effects of SSd in U87 cells were at least partly attributed to the depression of phosphatidylinositol 3­kinase/Akt and ERK protein expression levels, and activation of JNK and caspase-3 expression.