Overexpression of low-density lipoprotein receptor prevents neurotoxic polarization of astrocytes via inhibiting NLRP3 inflammasome activation in experimental ischemic stroke

Neurotoxic astrocytes are a promising therapeutic target for the attenuation of cerebral ischemia/reperfusion injury.Low-density lipoprotein receptor,a classic cholesterol regulatory receptor,has been found to inhibit NLR family pyrin domain containing protein 3(NLRP3)inflammasome activation in neurons following ischemic stroke and to suppress the activation of microglia and astrocytes in individuals with Alzheimer’s disease.However,little is known about the effects of low-density lipoprotein receptor on astrocytic activation in ischemic stroke.To address this issue in the present study,we examined the mechanisms by which low-density lipoprotein receptor regulates astrocytic polarization in ischemic stroke models.First,we examined low-density lipoprotein receptor expression in astrocytes via immunofluorescence staining and western blotting analysis.We observed significant downregulation of low-density lipoprotein receptor following middle cerebral artery occlusion reperfusion and oxygen-glucose deprivation/reoxygenation.Second,we induced the astrocyte-specific overexpression of low-density lipoprotein receptor using astrocyte-specific adeno-associated virus.Low-density lipoprotein receptor overexpression in astrocytes improved neurological outcomes in middle cerebral artery occlusion mice and reversed neurotoxic astrocytes to create a neuroprotective phenotype.Finally,we found that the overexpression of low-density lipoprotein receptor inhibited NLRP3 inflammasome activation in oxygen-glucose deprivation/reoxygenation injured astrocytes and that the addition of nigericin,an NLRP3 agonist,restored the neurotoxic astrocyte phenotype.These findings suggest that low-density lipoprotein receptor could inhibit the NLRP3-meidiated neurotoxic polarization of astrocytes and that increasing low-density lipoprotein receptor in astrocytes might represent a novel strategy for treating cerebral ischemic stroke.

Piezo1 as a potential player in intracranial hemorrhage:From perspectives on biomechanics and hematoma metabolism

Intracranial hemorrhage(ICH)causes numerous neurological deficits and deaths worldwide each year,leaving a significant health burden on the public.The pathophysiology of ICH is complicated and involves both primary and secondary injuries.Hematoma,as the primary pathology of ICH,undergoes metabolism and triggers biochemical and biomechanical alterations in the brain,leading to the secondary injury.Past endeavors mainly aimed at biochemical-initiated mechanisms for causing secondary injury,which have made limited progress in recent years,although ICH itself is also highly biomechanics-related.The discovery of the mechanically-activated cation channel Piezo1 provides a new avenue to further explore the mechanisms underlying the secondary injury.The current article reviews the structure and gating mechanisms of Piezo1,its roles in the physiology/pathophysiology of neurons,astrocytes,microglia,and bone-marrow-derived macrophages,and especially its roles in erythrocytic turnover and iron metabolism,revealing a potential interplay between the biomechanics and biochemistry of hematoma in ICH.Collectively,these advances provide deeper insights into the secondary injury of ICH and lay the foundations for future research.

A Novel Method for Cross-Subject Human Activity Recognition with Wearable Sensors

Human Activity Recognition (HAR) is an important way for lower limb exoskeleton robots to implement human-computer collaboration with users. Most of the existing methods in this field focus on a simple scenario recognizing activities for specific users, which does not consider the individual differences among users and cannot adapt to new users. In order to improve the generalization ability of HAR model, this paper proposes a novel method that combines the theories in transfer learning and active learning to mitigate the cross-subject issue, so that it can enable lower limb exoskeleton robots being used in more complex scenarios. First, a neural network based on convolutional neural networks (CNN) is designed, which can extract temporal and spatial features from sensor signals collected from different parts of human body. It can recognize human activities with high accuracy after trained by labeled data. Second, in order to improve the cross-subject adaptation ability of the pre-trained model, we design a cross-subject HAR algorithm based on sparse interrogation and label propagation. Through leave-one-subject-out validation on two widely-used public datasets with existing methods, our method achieves average accuracies of 91.77% on DSAD and 80.97% on PAMAP2, respectively. The experimental results demonstrate the potential of implementing cross-subject HAR for lower limb exoskeleton robots.

Research and Analysis of Grammatical Error Correction Technology for Chinese Documents

With the widespread use of Chinese globally, the number of Chinese learners has been increasing, leading to various grammatical errors among beginners. Additionally, as domestic efforts to develop industrial information grow, electronic documents have also proliferated. When dealing with numerous electronic documents and texts written by Chinese beginners, manually written texts often contain hidden grammatical errors, posing a significant challenge to traditional manual proofreading. Correcting these grammatical errors is crucial to ensure fluency and readability. However, certain special types of text grammar or logical errors can have a huge impact, and manually proofreading a large number of texts individually is clearly impractical. Consequently, research on text error correction techniques has garnered significant attention in recent years. The advent and advancement of deep learning have paved the way for sequence-to-sequence learning methods to be extensively applied to the task of text error correction. This paper presents a comprehensive analysis of Chinese text grammar error correction technology, elaborates on its current research status, discusses existing problems, proposes preliminary solutions, and conducts experiments using judicial documents as an example. The aim is to provide a feasible research approach for Chinese text error correction technology.

Adaptive Robust Control with Leakage-Type Control Law for Trajectory Tracking of Exoskeleton Robots

This paper investigates the trajectory following problem of exoskeleton robots with numerous constraints. However, as a typical nonlinear system with variability and parameter uncertainty, it is difficult to accurately achieve the trajectory tracking control for exoskeletons. In this paper, we present a robust control of trajectory tracking control based on servo constraints. Firstly, we consider the uncertainties (e.g., modelling errors, initial condition deviations, structural vibrations, and other unknown external disturbances) in the exoskeleton system, which are time-varying and bounded. Secondly, we establish the dynamic model and formulate a close-loop connection between the dynamic model and the real world. Then, the trajectory tracking issue is regarded as a servo constraint problem, and an adaptive robust control with leakage-type adaptive law is proposed with the guaranteed Lyapunov stability. Finally, we conduct numerical simulations to verify the performance of the proposed controller.

CPG Human Motion Phase Recognition Algorithm for a Hip Exoskeleton with VSA Actuator

Due to the dynamic stiffness characteristics of human joints, it is easy to cause impact and disturbance on normal movements during exoskeleton assistance. This not only brings strict requirements for exoskeleton control design, but also makes it difficult to improve assistive level. The Variable Stiffness Actuator (VSA), as a physical variable stiffness mechanism, has the characteristics of dynamic stiffness adjustment and high stiffness control bandwidth, which is in line with the stiffness matching experiment. However, there are still few works exploring the assistive human stiffness matching experiment based on VSA. Therefore, this paper designs a hip exoskeleton based on VSA actuator and studies CPG human motion phase recognition algorithm. Firstly, this paper puts forward the requirements of variable stiffness experimental design and the output torque and variable stiffness dynamic response standards based on human lower limb motion parameters. Plate springs are used as elastic elements to establish the mechanical principle of variable stiffness, and a small variable stiffness actuator is designed based on the plate spring. Then the corresponding theoretical dynamic model is established and analyzed. Starting from the CPG phase recognition algorithm, this paper uses perturbation theory to expand the first-order CPG unit, obtains the phase convergence equation and verifies the phase convergence when using hip joint angle as the input signal with the same frequency, and then expands the second-order CPG unit under the premise of circular limit cycle and analyzes the frequency convergence criterion. Afterwards, this paper extracts the plate spring modal from Abaqus and generates the neutral file of the flexible body model to import into Adams, and conducts torque-stiffness one-way loading and reciprocating loading experiments on the variable stiffness mechanism. After that, Simulink is used to verify the validity of the criterion. Finally, based on the above criterions, the signal mean value is removed using feedback structure to complete the phase recognition algorithm for the human hip joint angle signal, and the convergence is verified using actual human walking data on flat ground.

Predictive performance of'Diprifusor'TCI system in patients during upper abdominal surgery under propofol/fentanyl anesthesia

Objective:To evaluate the predictive performance of‘Diprifusor’TCI(target-controlled infusion)system for its betterapplication in clinical anesthesia.Methods:The predictive performance of a‘Diprifusor’TCI system was investigated in 27Chinese patients(16 males and 11 females)during upper abdominal surgery under total intravenous anesthesia(TIVA)withpropofol/fentanyl.Measnred arterial propofol concentrations were compared with the values predicted by the TCI infusion system.Performance was determined by the median performance error(MDPE),the median absolute performance error(MDAPE),thedivergence(the percentage change of the absolute PE with time),and the wobble(the median absolute deviation of each PE fromthe MDPE).Results:The median(range)values of 14.9%(-21.6%～42.9%)for MDPE,23.3%(6.9%～62.5%)for MDAPE,-1.9%h^(-1)(-32.7%～23.0% h^(-1))for divergence,and 18.9%(4.2%～59.6%)for wobble were obtained from 227 samples from all patients.For the studied population,the PE did not increase with time but with increasing target propofol concentration,particularly fol-lowing induction.Conclusions:The control of depth of anaesthesia was good in all patients undergoing upper abdominal surgicaloperation and the predictive performance of the‘Diprifusor’target controlled mthsion system was considered acceptable forclinical purposes.But the relatively bigger wobble showed that the pharmacokinetic model is not so suitable and requires im-provement.

New therapeutic options for persistent low-level viremia in patients with chronic hepatitis B virus infection:Increase of entecavir dosage

Chronic hepatitis B virus(HBV)infection(CHB)is a public health concern worldwide.Current therapies utilizing nucleos(t)ide analogs(NA)have not resulted in a complete cure for CHB.Furthermore,patients on long-term NA treatment often develop low-level viremia(LLV).Persistent LLV,in addition to causing the progression of liver disease or hepatocellular carcinoma,may shed light on the current plight of NA therapy.Here,we review the literature on LLV,NA treatment,and various doses of entecavir to find a strategy for improving the efficacy of this antiviral agent.For LLV patients,three therapeutic options are available,switching to another antiviral monotherapy,interferon-αswitching therapy,and continuing monotherapy.In real-world clinical practice,entecavir overdose has been used in antiviral therapy for CHB patients with NA refractory and persistent LLV,which encouraged us to conduct further in-depth literature survey on dosage and duration related entecavir studies.The studies of pharmacodynamics and pharmacokinetics show that entecavir has the maximal selected index for safety,and has great potential in inhibiting HBV replication,in all of the NAs.In the particular section of the drug approval package published by the United States Food and Drug Administration,entecavir doses 2.5-20 mg/d do not increase adverse events,and entecavir doses higher than 1.0 mg/d might improve the antiviral efficacy.The literature survey led us to two suggestions:(1)Increasing entecavir dose to 1.0 mg/d for the treatment of NA naïve patients with HBV DNA>2×106 IU/mL is feasible and would provide better prognosis;and(2)Further research is needed to assess the long-term toxic effects of higher entecavir doses(2.5 and 5.0 mg/d),which may prove beneficial in treating patients with prior NA treatment,partial virological response,or LLV state.

Localization and potential function of androgen receptor in rat salivary gland

Aim： To investigate the localization and quantity of androgen receptor （AR） in the salivary glands of rats with further analysis on the effect of castration. Methods： Sixty male Wistar rats, aged 30-60 days, were randomly divided into three groups （castrated, sham-operated and normal controls） with 20 rats in each group. The rats in the castrated group were castrated and the submaxillary glands were removed after 1 week. The salivary glands of the rats in the sham-operated and the normal control groups were also removed. Parts of the salivary glands were fixed for immuohistochemistry and in situ hybridization assays. Other parts were used for Western blot. Results： AR immunoreactivity in the three groups was localized in the glandular epithelial cells of the serous acinus and the glandular duct of the salivary gland, mainly in the nuclei. AR mRNA hybridization signals in the salivary glands of the castrated group were mainly distributed in the epithelial cells of the convoluted and secretary ducts; AR mRNA in the sham-operated and the normal control groups were found in the epithelial cells of the convoluted, the secretary and the excretory ducts. The quantity of AR in the salivary glands was decreased significantly in the castrated rats compared with the sham-operated and the normal controls. Moreover, epidermal growth factor （EGF） secreted by the salivary glands was also decreased in the castrated rats. Conclusion： Castration appears to affect the production of AR in the salivary gland and the distribution of the AR mRNA and could further affect the function of the salivary gland. The changes of AR and the distribution of AR mRNA may play an important role in the interactions between the testes and the salivary gland. （Asian J Androl 2005 Sep; 7： 295-301）

Follicle-stimulating hormone autoantibody is involved in idiopathic spermatogenic dysfunction

Aim： To detect the anti-follicle-stimulating hormone （FSH） antibody in idiopathic infertile patients and fertile subjects in order to determine the role of this antibody in patients with spermatogenic dysfunction. Methods： The anti-FSH antibody in serum was detected by an enzyme-linked immunosorbent assay （ELISA）. The functional and structural integrity of the sperm membrane was evaluated with hypo-osmotic swelling （HOS） test and the ultrastructure of the spermatozoa was investigated by transmission electron microscopy （TEM）. Results： The extent of positive FSH antibody in the patients with oligozoospermia and/or asthenozoospermia was significantly higher than that in the fertile subjects and infertile patients with normal sperm concentration and motility, but it was significantly lower than that in the patients with azoospermia. The extent of anti-FSH antibody in the patients with azoospermia was significantly greater than that in patients with oligospermia and/or asthenospermia, infertile people with normal sperm density and motility and fertile people. The hypo-osmotic swelling test showed that the percentage of HOS-positive spermatozoa （swollen） was 45.1% ±3.5% in the FSH antibody-positive group and 59.1% ± 6.2% in the FSH antibody-negative control group. The percentage of functional membrane damage to spermatozoa was significantly higher in the anti- FSH antibody-positive group than in the control group. TEM showed that the outer acrosomal membrane was located far from the nucleus, and detachment of the acrosome was found in the FSH autoantibody-positive group. Conclusion： These data suggest that the presence of anti-FSH antibody is strongly correlated with the sperm quantity and quality in idiopathic male infertility. Anti-FSH antibody may be an important factor causing spermatogenic dysfunction and infertility.

Ultrasound-guided Open Nephron Sparing Surgery without Renal Artery Occlusion for Central Renal Tumors

From January 2008 to January 2013, 11 patients with central renal tumors underwent ultrasound-guided open nephron sparing surgery（ONSS） without renal artery occlusion. We removed the lesions, and the cut edges of the tumors were negative. Thus, we deduced that ultrasound-guided ONSS is suitable for the cases with obscure tumor boundary or multiple lesions. It could achieve the purpose of thoroughly removing lesions, as well as to expand the application range of nephron sparing surgery.

Liver injury changes the biological characters of serum small extracellular vesicles and reprograms hepatic macrophages in mice

BACKGROUND Serum small extracellular vesicles(sEVs)and their small RNA(sRNA)cargoes could be promising biomarkers for the diagnosis of liver injury.However,the dynamic changes in serum sEVs and their sRNA components during liver injury have not been well characterized.Given that hepatic macrophages can quickly clear intravenously injected sEVs,the effect of liver injury-related serum sEVs on hepatic macrophages deserves to be explored.AIM To identify the characteristics of serum sEVs and the sRNAs during liver injury and explore their effects on hepatic macrophages.METHODS To identify serum sEV biomarkers for liver injury,we established a CCL4-induced mouse liver injury model in C57BL/6 mice to simulate acute liver injury(ALI),chronic liver injury(CLI)and recovery.Serum sEVs were obtained and characterized by transmission electron microscopy and nanoparticle tracking analysis.Serum sEV sRNAs were profiled by sRNA sequencing.Differentially expressed microRNAs(miRNAs)were compared to mouse liver-enriched miRNAs and previously reported circulating miRNAs related to human liver diseases.The biological significance was evaluated by Ingenuity Pathway Analysis of altered sEV miRNAs and conditioned cultures of ALI serum sEVs with primary hepatic macrophages.RESULTS We found that both ALI and CLI changed the concentration and morphology of serum sEVs.The proportion of serum sEV miRNAs increased upon liver injury,with the liver as the primary contributor.The altered serum sEV miRNAs based on mouse studies were consistent with human liver disease-related circulating miRNAs.We established serum sEV miRNA signatures for ALI and CLI and a panel of miRNAs(miR-122-5p,miR-192-5p,and miR-22-3p)as a common marker for liver injury.The differential serum sEV miRNAs in ALI contributed mainly to liver steatosis and inflammation,while those in CLI contributed primarily to hepatocellular carcinoma and hyperplasia.ALI serum sEVs decreased both CD86 and CD206 expression in monocyte-derived macrophages but increased CD206 expression in resident macrophages in vitro.CONCLUSION Serum sEVs acquired different concentrations,sizes,morphologies and sRNA contents upon liver injury and could change the phenotype of liver macrophages.Serum sEVs therefore have good diagnostic and therapeutic potential for liver injury.

Cerebrospinal fluid and plasma propofol concentration during total intravenous anaesthesia of patients undergoing elective intracranial tumor removal

Objective： The aim of this paper is to compare the propofol concentration in plasma and cerebrospinal fluid （CSF） in patients scheduled for intracranial tumor removal and anaesthetized using propofol as part of a total intravenous anaesthesia technique. Methods： Twenty-seven patients （ASA Ⅰ-Ⅱ） scheduled for elective intracranial tumor removal were studied. Anesthesia was induced with 2 mg/kg propofol for 5 min and infused at 10 mg/（kg·h） for 5 min and then stopped. CSF and arterial blood were collected simultaneously before infusion of propofol and at different time points after infusion ofpropofol according to bispectral index （BIS） values. Concentrations of propofol in plasma and CSF were measured by HPLC with fluorescence detection. The correlation coefficient and regression equation between plasma and CSF concentration of propofol were worked out by linear simple regression. Results： The propofol CSF concentration that we measured was 1.46% of the plasma concentration. The coefficient of relation between plasma and CSF concentration was 76.7%. Conclusions： The propofol CSF concentration was positively correlated with and much lower than the plasma concentration. Discrepancies may result from high plasma protein binding of propofol, intracranial pathology and sampling volume.

Protective effects of ferulic acid against ionizing radiation-induced oxidative damage in rat lens through activating Nrf2 signal pathway

AIM: To examine the protection of ferulic acid(FA) against ionizing radiation(IR)-induced lens injury in rats, as well as the underlying mechanisms.METHODS: FA(50 mg/kg) was administered to rats for 4 consecutive days before they were given 10 Gy γ-radiation, as well as for 3 consecutive days afterward. Two weeks after radiation, the eye tissues were collected. Histological alterations were evaluated by hematoxylineosin staining. Enzyme linked immunosorbent assay(ELISA) was utilized to assess the activities of glutathione reductase(GR) and superoxide dismutase(SOD), as well as the levels of glutathione(GSH) and malondialdehyde(MDA) in the lenses. The protein and m RNA levels of Bcl-2, caspase-3, Bax, heme oxygenase-1(HO-1), and glutamatecysteine ligase catalytic subunit(GCLC) were quantified using Western blot and quantitative reverse transcription polymerase chain reaction, respectively. With nuclear extracts, the nuclear factor erythroid-2 related factor(Nrf2) protein expressions in the nuclei were also measured.RESULTS: Rats exposed to IR showed lens histological alterations which could be alleviated by FA. FA treatment reversed apoptosis-related markers in IR-induced lens, as evidenced by lower levels of Bax and caspase-3 and higher level of Bcl-2. Furthermore, IR induced oxidative damage manifested by decreased GSH level, increased MDA level, and decreased SOD and GR activities. FA boosted nuclear translocation of Nrf2 and increased the expressions of HO-1 and GCLC to inhibit oxidative stress, as evidenced by an increase in GSH, a decrease in MDA, and an increase in GR and SOD activities.CONCLUSION: FA may work well in preventing and treating IR-induced cataract through promoting the Nrf2 signal pathway to attenuate oxidative damage and cell apoptosis.

Improving antibody affinity through in vitro mutagenesis in complementarity determining regions

High-affinity antibodies are widely used in diagnostics and for the treatment of human diseases.However,most antibodies are isolated from semi-synthetic libraries by phage display and do not possess in vivo affinity maturation,which is triggered by antigen immunization.It is therefore necessary to engineer the affinity of these antibodies by way of in vitro assaying.In this study,we optimized the affinity of two human monoclonal antibodies which were isolated by phage display in a previous related study.For the 42A1 antibody,which targets the liver cancer antigen glypican-3,the variant T57H in the second complementarity-determining region of the heavy chain(CDR-H2)exhibited a 2.6-fold improvement in affinity,as well as enhanced cell-binding activity.For the I4A3 antibody to severe acute respiratory syndrome coronavirus 2,beneficial single mutations in CDR-H2 and CDR-H3 were randomly combined to select the best synergistic mutations.Among these,the mutation S53P-S98T improved binding affinity(about 3.7 fold)and the neutralizing activity(about 12 fold)compared to the parent antibody.Taken together,single mutations of key residues in antibody CDRs were enough to increase binding affinity with improved antibody functions.The mutagenic combination of key residues in different CDRs creates additive enhancements.Therefore,this study provides a safe and effective in vitro strategy for optimizing antibody affinity.

Promoting chondrogenesis by targeted delivery to the degenerating cartilage in early treatment of osteoarthritis

Osteoarthritis(OA)is a highly incident total joint degenerative disease with cartilage degeneration as the primary pathogenesis.The cartilage matrix is mainly composed of collagen,a matrix protein with a hallmark triplehelix structure,which unfolds with collagen degradation on the cartilage surface.A collagen hybridizing peptide(CHP)is a synthetic peptide that binds the denatured collagen triple helix,conferring a potential diseasetargeting possibility for early-stage OA.Here,we constructed an albumin nanoparticle(An)conjugated with CHP,loaded with a chondrogenesis-promoting small molecule drug,kartogenin(KGN).The CHP-KGN-An particle exhibited sustained release of KGN in vitro and prolonged in vivo retention selectively within the degenerated cartilage in the knee joints of model mice with early-stage OA.Compared to treatment with KGN alone,CHP-KGN-An robustly attenuated cartilage degradation,synovitis,osteophyte formation,and subchondral bone sclerosis in OA model mice and exhibited a more prominent effect on physical activity improvement and pain alleviation.Our study showcases that targeting the degenerated cartilage by collagen hybridization can remarkably promote the efficacy of small molecule drugs and may provide a novel delivery strategy for earlystage OA therapeutics.

The effect of vitamin D on sperm motility and the underlying mechanism

Vitamin D deficiency is a common health issue around the world. We therefore evaluated the associations of semen quality with both serum and seminal plasma vitamin D levels and studied the mechanisms underlying these by incubating spermatozoa with 1,25(OH)2D in vitro. Two hun dred and twenty-two men were in eluded in our study. Vitamin D was detected using an electrochemilumi nesce nee method. Spermatozoa used for in vitro experiments were isolated by density gradient centrifugation. Positive relationships of serum 25(OH)D with semen volume and seminal plasma fructose were identified. Seminal plasma 25(OH)D level showed no relationship with serum 25(OH)D level, while it was inversely associated with sperm concentration and positively correlated with semen volume and sperm kinetic values. In vitro, sperm kinetic parameters in creased after in cubation with 1,25(OH)2D, especially upon in cubation for 30 min with it at a concen tration of 0.1 nmol l-1. Under these in cubation conditions, the upward migratio n of spermatozoa in creased remarkably with increasing ade nosine triphosphate (ATP) con centratio n. The concentrati on of cyclic ade nosine mono phosphate (cAMP) and the activity of protei n kinase A (PKA) were both elevated, and the PKA inhibitor, N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H89) reversed the in crease of ATP producti on. The conce ntrations of cytoplasmic calcium ions and n icotinamide adenine dinucleotide (NADH) were both enhanced, while mitochondrial calcium uniporter (MCU) inhibitor, Ruthenium 360 (Ru360) did not reverse the increase of ATP production. Therefore, seminal plasma vitamin D may be invoIved in regulating sperm motility, and 1,25(OH)2D may enhance sperm motility by promoting the synthesis of ATP both through the cAMP/PKA pathway and the in crease in in tracellular calcium ions.

Primary obturator foramen pregnancy： a case report and review of literature

Ectopic pregnancy, in which the fertilized ovum implants on any tissue other than the endometrium, is the most common life-threatening emergency in the first trimester of pregnancy. The incidence of ectopic pregnancy is 0.25%-1.0% of all pregnancies, and has increased greatly in the last few decades, from 4.5 per 1000 pregnancies in 1970 to an estimated 19.7 per 1000 pregnancies in 1992.^1

Sturge-Weber syndrome： a case report and review of literatures

Sturge-Weber syndrome （SWS）, or encephalo- trigeminal angiomatosis, is a rare, congenitalneurocutaneous syndrome characterized by unilateral facial cutaneous vascular malformation （nevus flammeus or port-wine stain） in association with ipsilateral leptomeningeal angiomatosis. Prevalence is approximately one per 50 000 live births. Males and females are equally affected and there is no racial bias. 1,2 The common clinical manifestations of SWS include progressive seizures, unilateral cutaneous vascular nevus following the ophthalmic divisions of the trigeminal nerve, ipsilateral glaucoma, contralateral hemiparesis, hemiatrophy, hemianopia and mental retardation. The radiographic hallmark of SWS is ＂tram-line＂ or gyriform calcifications usually involving the occipital and parietal lobes. Histologic studies have revealed that intracranial lesions of SWS display as leptomeningeal angiomatosis, and gyriform calcifications, neuronal loss, astrogliosis in underlying brain tissue.

Annexin A5 regulates Leydig cell testosterone production via ERK1/2 pathway

This study was to investigate the effect of annexin A5 on testosterone secretion from primary rat Leydig cells and the underlying mechanisms. Isolated rat Leydig ceils were treated with annexin A5. Testosterone production was detected by chemiluminescence assay. The protein and mRNA of Steroidogenic acute regulatory （STAR）, P450scc, 3β-hydroxysteroid dehydrogenase （3β-HSD）, 17β-hydroxysteroid dehydrogenase （17β-HSD）, and 17β-hydroxylase were examined by Western blotting and semi-quantitative RT-PCR, respectively. Annexin A5 significantly stimulated testosterone secretion from rat Leydig cells in dose- and time-dependent manners and increased mRNA and protein expression of STAR, P450scc, 3β-HSD, and 17β-HSD but not 17α-hydroxylase. Annexin A5 knockdown by siRNA significantly decreased the level of testosterone and protein expression of P450scc, 3β-HSD, and 17β-HSD. The significant activation of ERK1/2 signaling was observed at 5, 10, and 30 min after annexin A5 treatment. After the pretreatment of Leydig cells with ERK inhibitor PD98059 （50 μmol l^-1） for 20 min, the effects of annexin A5 on promoting testosterone secretion and increasing the expression of P450scc, 3β-HSD, and 17β-HSD were completely abrogated （P 〈 0.05）. Thus, ERK1/2 signaling is involved in the roles of annexin A5 in mediating testosterone production and the expression of P450scc, 3β-HSD, and 17β-HSD in Leydig cells.