Immunomodulatory roles of metalloproteinases in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic, autoimmune pathology characterized by persistent synovial inflammation and gradually advancing bone destruction. Matrix metalloproteinases (MMPs), as a family of zinc-containing enzymes, have been found to play an important role in degradation and remodeling of extracellular matrix (ECM). MMPs participate in processes of cell proliferation, migration, inflammation, and cell metabolism. A growing number of persons have paid attention to their function in inflammatory and immune diseases. In this review, the details of regulation of MMPs expression and its expression in RA are summarized. The role of MMPs in ECM remodeling, angiogenesis, oxidative and nitrosative stress, cell migration and invasion, cytokine and chemokine production, PANoptosis and bone destruction in RA disease are discussed. Additionally, the review summarizes clinical trials targeting MMPs in inflammatory disease and discusses the potential of MMP inhibition in the therapeutic context of RA. MMPs may serve as biomarkers for drug response, pathology stratification, and precision medicine to improve clinical management of rheumatoid arthritis.

[Ginkgo biloba Ketone Ester Tablets with different release rates prepared by fused deposition modeling 3D printing technology].

The present study prepared a new type of Ginkgo biloba ketone ester(GBE50) preparation from polyethylene glycol and croscarmellose sodium with good biocompatibility and a certain viscosity by fused deposition modeling(FDM)-type 3D printing technique. Firstly, a cylindrical 3D printing model with a diameter of 9.00 mm and a height of 4.50 mm was established. Subsequently, the 3D-GBE50 preparations with three paths(concentric, zigzag, and grid), different layer heights, and different filling gaps were designed and prepared after the optimization of the proportions of excipients. The morphology, size, chemical properties, and dissolution activity of the 3D-GBE50 preparations were fully characterized and investigated. The results showed that 3D-GBE50 preparations had smooth appearance, clear texture, standard friability, good thermal stability, and stable chemical properties. Moreover, the printing path, layer height, and filling gap were directly related to the release rate of 3D-GBE50 preparations. The dissolution of 3D-GBE50 tablets with zigzag printing path was the fastest, while the dissolution rates of 3D-GBE50 tablets with concentric circle and grid-shaped printing paths were slower than that of commercially available G. biloba Ketone Ester Tablets. In addition, the dissolution of 3D-GBE50 tablets was faster with higher layer height and wider filling gap. As revealed by the results, th FDM-type 3D printing technique can flexibly regulate the drug release activity via controlling the printing parameters, providing effective ideas and methods for the pre-paration of personalized pharmaceutical preparations.

Efficacy of combined naikan and morita therapies on psychological distress and posttraumatic growth in Chinese patients with advanced cancer: A randomized controlled trial.

BACKGROUND: Advanced cancer (AC) patients experience serious physical and psychological problems with the disease progression. When approaching the end of life, these patients have to cope with not only the bodily illness, but also the spiritual crisis. Conventional psychological treatments reduce distress to a certain extent, but for patients with AC, especially when they face progressive illness and are approaching death, their psychological problems are complex, and no simple solutions are in sight. Therefore, we designed this study to evaluate the efficacy of the combined Naikan therapy (NT) and Morita therapy (MT) on psychological distress and posttraumatic growth in patients with AC. METHOD: One hundred thirty patients newly diagnosed with AC were allocated randomly into treatment (n = 65) and control (n = 65) groups. Patients in the treatment group received combined NT and MT for 7 consecutive weeks, while the control group received normal medical treatments without NT and MT. Patients were assessed before and after the therapies. The primary outcome measures include distress thermometer (DT) and posttraumatic growth, and the secondary outcome measure contains the list of distress problems. RESULTS: At the post-treatment stage, the treatment group displayed a decreased score of psychological distress as compared to that in the control group, which accompanied by a higher post-traumatic growth total score and subscale scores in relationship to others, new possibilities, personal strength, spiritual changes, and appreciation of life. A significant decrease in fear, sleeping difficulty/insomnia, nervousness/anxiety, and loss of appetite was also observed in the treatment group. CONCLUSION: The results proved that the combined Naikan and Morita therapies decreased the psychological distress and improved the posttraumatic growth of the patients with AC. TRIAL REGISTRATION: ChiCTR1900026691.

[Effects of late sowing of two season crops on annual yield and resource use efficiency in rice-wheat double cropping system]. / 稻麦两熟系统适期晚播对周年产量和资源利用效率的影响.

The unreasonable resource allocation and lower resource use efficiency for rice-wheat double cropping system in Jianghuai region resulted from climate change severely limit the coordinated development of annually high yield and high efficiency crops. Optimizing seasonal resource allocation through sowing date adjustment is an important way to tap the annual high-yield potential and improve resource use efficiency. To quantify the effects of sowing date of rice and wheat on annual yield and resource allocation and utilization efficiency, field experiments were conducted in 2013-2015. Results showed that compared with the conventional rice-wheat cropping system (T2), the two seasons appropriate late-cast cropping system (T3) could coordinate resource allocation in the two seasons through the sowing date adjustment, and transfer the redundant radiation and heat resources in the wheat season to the rice. The distribution rate of accumulated temperature, radiation and rainfall resources for T3 were: rice season accounted for 60.5%, 46.5% and 56.7%, wheat season accounted for 36.3%, 50.0% and 40.9%, and the ratio between two seasons was 1.67, 0.94 and 1.39, respectively. Rice yield and its proportion of annual production were significantly increased. The wheat yield was significantly decreased, with the variation range being smaller than that of rice. The total annual yield was increased by 336.3 kg·hm-2 as compared with T2. The temperature, radiation and rainfall production efficiency for rice in T3 were increased by 9.8%, 5.6% and 8.3% in compared to T2, respectively. There was no significant difference in the climate resource utilization efficiency of wheat season. The annual resource production efficiency of T3 was increased by 4.8%, 3.1% and 6.0% over the T2, respectively. Earlier (T1) or latest sowing (T4) of two seasons cropping system was not appropriate for annual yield formation and resource utilization. In summary, improving resource utilization efficiency in rice season is the key way to increase annual grain yield potential in Jianghuai region. The results provided theoretical and practical bases for the excavation of yield potential of the regional annual cropping system and the adjustment of planting structure.

Role of miR-199a-5p in osteoblast differentiation by targeting TET2.

OBJECTIVE: miR-199a-5p was increased during osteoblast differentiation, which may target and regulate TET2, a gene attracted a lot of attention in the osteoblast differentiation in the past few years. However, the role of miR-199a-5p in osteoblast differentiation by targeting TET2 is not established. METHODS: The correlation between miR-199a-5p and TET2 was verified through dual luciferase reporter assay, and their expressions in human bone marrow stromal cells (hBMSCs) during the osteoblast differentiation were detected. hBMSCs were transfected with TET2 siRNA, miR-199a-5p mimic or/and TET2 CRISPR activation plasmid., and then prepared for the induction of osteoblast differentiation, followed by alkaline phosphatase (ALP) and alizarin red staining, qRT-PCR and Western blotting. In vivo, ovariectomized (OVX) mice were injected with agomir-miR-199a-5p, antagomiR-199a-5p or/and TET2 siRNA to calculate the BMD and BV/TV ratio of mice, as well as to measure the expressions of osteogenesis-related genes in bone tissues. RESULTS: A gradual increase of miR-199a-5p was observed in hBMSCs during the induction of osteoblast differentiation, while TET2 expression was decreased. Besides, miR-199a-5p was reduced in the bone tissue of OVX mice, while TET2 was up-regulated. In addition, overexpression of miR-199a-5p and inhibition of TET2 augmented ALP activity in hBMSCs, with the enhanced calcification and the up-regulated expressions of Runx2, OSX and OCN, which also increased the quality of bone in OVX mice accompanying the enhancement BV/TV ratio, BMD and osteogenesis-related genes. CONCLUSION: MiR-199a-5p may promote the osteoblast differentiation and prevent OVX-induced osteoporosis by targeting TET2.

Benefits and Risks of Subsection Laminectomy with Pedicle Screw Fixation for Ossification of the Ligamentum Flavum of the Thoracic Spine: A Retrospective Study of 30 Patients.

BACKGROUND This study aimed to evaluate the effectiveness of subsection laminectomy with pedicle screw fixation (SLPF) for the treatment of ossification of the ligamentum flavum of the thoracic spine. MATERIAL AND METHODS Thirty patients (age, 40-71 years) with ossification of the ligamentum flavum of the thoracic spine underwent SLPF (13 men, 17 women). Operative time, intraoperative blood loss, preoperative and postoperative change in thoracic kyphosis, and perioperative complications were recorded. The Japanese Orthopedic Association (JOA) score for severity of myelopathy and the American Spinal Injury Association (ASIA) motor and sensory impairment scale were used before and after surgery. RESULTS Mean operative time for SLPF was 208.4±38.3 min and mean intraoperative blood loss was 689.3±171.7 ml. The mean JOA score significantly increased from 5.7±1.9 before surgery to 8.8±2.2 at one month after surgery and 9.3±2.7 at the last follow-up (P<0.01). Postoperative improvement in neurological function increased by 68.3±14.4%. The postoperative ASIA grades significantly improved compared with the preoperative grades (P<0.01). The mean local Cobb angle significantly decreased from 17.8±4.3° before surgery to 15.4±3.6° at one month after surgery and 15.8±3.8° at the last follow-up (P<0.01). Three patients (10%) had operative cerebrospinal fluid (CSF) leak. Postoperatively, one patient had neurological deterioration, two patients had deep venous thrombosis (DVT), and one patient developed a wound infection. CONCLUSIONS SLPF was an effective procedure for the treatment of ossification of the ligamentum flavum of the thoracic spine.

[Cloning and expression of SmDXS2 gene in Swertia mussotii].

1-deoxy-D-xylulose-5-phosphate synthase2(DXS2) is the first key enzyme of the MEP pathway,which plays an important role in terpene biosynthesis of plants. According to the data of Swertia mussotii transcriptome, DXS2 gene(Gen Bank number MH535905) was cloned and named as Sm DXS2. The bioinformatics results showed that Sm DXS2 has no intron,with a 2 145 bp open reading frame encoding a polypeptide of 714 amino acids. They are belonging to 20 kinds of amino acids,and the most abundant amino acids include Ala,Gly and Trp. The predicted protein molecular weight was 76. 91 k Da and its theoretical isoelectric point(p I) was6. 5,which belonging to a hydrophilic protein. α-Helix and loop were the major motifs of predicted secondary structure of DXS2. The three function domains are TPP＿superfamily,Transket＿pyr＿ superfamily and Transketolase＿C superfamily,respectively. The Sm DXS2 protein shared high identity with other DXS2 proteins of plants. Phylogenetic analysis showed that Sm DXS2 protein is grouped with the gentian DXS2 protein. The recombinant protein of Sm DXS2 gene in Escherichia coli was approximately 92. 00 k Da(containing sumo-His tag protein 13 k Da),which was consistent with the anticipated size.This work will provide a foundation for further functional research of Sm DXS2 protein and increasing the product of iridoid compound by genetic engineering in S. mussotii.

[Effects of nitrogen fertilizer reduction management on photosynthesis and chlorophyll fluorescence characteristics of sweetpotato]. / å‡æ°®è¿ç­¹å¯¹ç”˜è–¯å ‰åˆä½œç”¨å’Œå¶ç»¿ç´ è§å ‰ç‰¹æ€§çš„å½±å“.

Crop productivity depends on photosynthetic source capacity. Appropriate nitrogen (N) fertilizer management is beneficial for improving growth, photosynthetic capacity and thereby increasing crops yield. A two-year pot experiment was conducted with four N treatments, i.e., conventional basal application 100 kg N·hm-2 as control (FP), a total of 80 kg N·hm-2 applied either 100% at basal application (JS), 100% at tuber initiation stage (35 d after transplant, KS), 50% at basal application and 50% at tuber initiation stage (35 d after transplant, FS), to examine the effects of reduced nitrogen fertilizer combined with application methods on the photosynthesis and chlorophyll fluorescence characteristics of sweetpotato (Ipomoea batatas) during summer 2016 and 2017. The results showed that the conventional basal application of a reduced N rate decreased photosynthesis of sweetpotato during the final growth phases compared to conventional application, dressing application relatively delayed late-season leaf senescence as indicated by the increased net photosynthetic rate (Pn), stomatal conductance (gs), intercellular CO2 concentration (Ci) and chlorophyll (Chl a+b) content during tuber expansion period. Split application of N fertilizer had noticeably higher Pn, gs, Ci and Chl a+b than other treatments. Furthermore, split application of N fertilizer had a significantly higher photochemical efficiency of photosystem 2 (Fv/Fm), quantum yield of electron transport (ΦPS2), and photochemical quenching co-efficient (qP), but lower initial fluorescence (Fo) and non-photochemical quenching coefficient (NPQ) during tuber expansion period. The improved photosynthesis by split N was due to both increased Fv/Fm with higher electron transfer rate and reduced thermal dissipation of light energy in the tuber expansion period. Results were consistent between two sweetpotato cultivars across years. The results indicated that one-time fertilization at basal or tuber initiation stage were not conducive to sweetpotato leaf photosynthesis. The split N application was more beneficial in terms of delaying late-season leaf senescence, extending leaf function period, enhancing photosynthesis and biomass production under reduced N application rate, which would be beneficial for sweetpotato yield.

Protective effects of celastrol against γ irradiation-induced oxidative stress in human umbilical vein endothelial cells.

High-dose ionizing radiation can cause harmful effects on the cardiovascular system. Notably, endothelial cells are critical targets in radiation-induced damage. γ radiation exerts its biological effects through the radiolysis of water, which further generates ROS and induces lipid peroxidation and DNA damage. The present study aimed to evaluate the potential protective effects of celastrol against γ radiation-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). HUVECs were exposed to γ radiation at different doses with or without celastrol treatment. Cell viability and cytotoxicity, migratory ability, ROS production, lipid peroxidation, oxidative DNA damage and antioxidative enzyme levels were evaluated in HUVECs at 24 h post-irradiation. It was observed that HUVECs exhibited decreased cell viability, increased cytotoxicity and a decreased migratory ability after exposure to 20-Gy γ radiation. Celastrol treatment concentration-dependently reversed these effects. γ irradiation was also demonstrated to increase the production of ROS, enhance lipid peroxidation and oxidative DNA damage and decrease the levels of SOD, catalase, GST and GPx in HUVECs. These detrimental effects were blocked by treatment with celastrol for 24 h. These data suggested that celastrol not only attenuated γ radiation-induced cytotoxicity, but also effectively blocked oxidative stress in HUVECs. As an antioxidant agent, celastrol may have potential protective effects in HUVECs against γ irradiation-induced injury.

Effects of sealing the intramedullary femoral canal in total knee arthroplasty: A randomized study.

BACKGROUND: This study investigates the clinical effects of sealing the femoral canal by intramedullary alignment instrumentation in total knee arthroplasty (TKA). METHODS: One hundred twenty consecutive patients with knee osteoarthritis, who underwent unilateral TKA, were enrolled in the study and equally randomized into 2 groups: the sealing group and the control group. In the sealing group, the femoral canal was sealed with autogenous bone and cement using intramedullary alignment instrumentation, while the femoral hole was left open for patients in the control group. Blood loss, hemoglobin (Hb) reduction, and other parameters were recorded, as well as the duration of hospital stay and complications. The Hospital for Special Surgery (HSS) knee score was used to assess knee function at the final follow-up appointment. RESULTS: The calculated blood loss, hidden blood loss, transfusion requirements, drainage volume, and Hb reduction measurements were significantly different (P < .05) between the 2 groups. There were no significant differences in the surgery time, intraoperative blood loss, length of hospital stay, HSS score or complications between the 2 groups (P > .05). CONCLUSIONS: Sealing the intramedullary canal with autologous bone and a cement plug is an effective method for reducing blood loss and decreasing blood transfusion requirements during TKA procedures that have increasing complication rates.

Cathepsin L is involved in 6-hydroxydopamine induced apoptosis of SH-SY5Y neuroblastoma cells.

Our previous study reported that cathepsin L may contribute to the death of dopaminergic neurons in rodent model of Parkinson's disease (PD). In this study we detected the changes in the expression of lysosomal cathepsin L in cellular models of PD. In human neuroblastoma SH-SY5Y cells, treatment with 6-hydroxydopamine caused an increase in cathepsin L immunoreactivity in the cytoplasm and an increased production of the active form of cathepsin L. The contribution of cathepsin L to 6-OHDA-induced NF-κB activation and death of SH-SY5Y neuroblastoma cells were evaluated with an irreversible inhibitor of cathepsin L, Z-FY(t-Bu)-DMK. 6-OHDA-induced IκB-α degradation, NF-κB p65 nuclear translocation, p53 and PUMA expression were partially blocked by Z-FY(t-Bu)-DMK. In addition, Z-FY(t-Bu)- DMK modulated the Bcl-2 family levels, and suppressed caspase-3 activation. These data indicate that cathepsin L may be involved in 6-OHDA-induced apoptosis and Parkinsonian neurodegeneration.

Application of labeled radioimmunoimaging tracing in detecting pulmonary embolism in rabbits after bone cement perfusion and relevant treatment effects.

BACKGROUND: During the process of bone cement joint replacement, some patients show a series of complications, such as a sudden drop in blood pressure or dyspnea. The cause of the complication is considered to be due to emboli caused by the femur prosthesis insertion. The purpose of the present study was to detect the pulmonary embolism in rabbits after bone cement perfusion by radioimmunoimaging, and to explore its protective measures. METHODS: Forty rabbits, 2.5 - 3.0 kg weight, were randomly assigned to four groups, with ten rabbits in each group. Group I (no intervention): Bone cement perfusion was done after medullary cavity reaming and pressurizing. Group II (epinephrine hydrochloride intervention): The medullary cavity was rinsed with a 1:10 000 normal saline-diluted epinephrine hydrochloride solution followed by bone cement perfusion after medullary cavity reaming and pressurizing. Group III (fibrin sealant intervention): The medullary cavity was precoated with fibrin sealant followed by bone cement perfusion after medullary cavity reaming and pressurizing. Group IV (blank control group): The medullary cavity was not perfused with bone cement after reaming. In each group, the rabbits underwent femoral head resection and medullary cavity reaming. Before bone cement perfusion, 2 ml of developing tracer was injected through the ear vein. Radionuclide imaging was performed at 60, 120, and 180 minutes after bone cement perfusion, and the pulmonary radioactivity in vivo was measured. The rabbits were immediately sacrificed, and the pulmonary tissue was removed and its radioactivity was measured in vitro. Pulmonary tissue was then fixed and the pulmonary embolism and the associated pathological changes were observed. RESULTS: The pulmonary radioactivity in vivo was measured at 60, 120, and 180 minutes after bone cement perfusion. The radioactivities of the four groups were 11.67 ± 2.16, 14.59 ± 2.92 and 18.43 ± 4.83 in group I; 8.37 ± 3.05, 10.35 ± 2.24 and 11.48 ± 2.96 in group II; 3.91 ± 1.19, 5.53 ± 2.95 and 7.25 ± 1.26 in group III; 1.04 ± 0.35, 1.14 ± 0.87 and 1.43 ± 0.97 in group IV. The radioactivities of groups I, II, III at 60, 120 and 180 minutes were significantly higher than group IV (P < 0.05). The pulmonary embolism could be detected. Pretreatment with epinephrine hydrochloride and fibrin sealant significantly decreased the pulmonary radioactivity in group II and group III, but it was still higher than in the group IV. CONCLUSIONS: Radioimmunoimaging is an alternative method for the dynamic observation of rabbit pulmonary embolism after bone cement perfusion. Radioimmunoimaging is the optional way to evaluate the effect of pretreatment with epinephrine hydrochloride or fibrin sealant on pulmonary embolism after bone cement perfusion.

Application of labeled radioimmunoimaging tracing in detecting pulmonary embolism in rabbits after bone cement perfusion and relevant treatment effects / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>During the process of bone cement joint replacement, some patients show a series of complications, such as a sudden drop in blood pressure or dyspnea. The cause of the complication is considered to be due to emboli caused by the femur prosthesis insertion. The purpose of the present study was to detect the pulmonary embolism in rabbits after bone cement perfusion by radioimmunoimaging, and to explore its protective measures.</p><p><b>METHODS</b>Forty rabbits, 2.5 - 3.0 kg weight, were randomly assigned to four groups, with ten rabbits in each group. Group I (no intervention): Bone cement perfusion was done after medullary cavity reaming and pressurizing. Group II (epinephrine hydrochloride intervention): The medullary cavity was rinsed with a 1:10 000 normal saline-diluted epinephrine hydrochloride solution followed by bone cement perfusion after medullary cavity reaming and pressurizing. Group III (fibrin sealant intervention): The medullary cavity was precoated with fibrin sealant followed by bone cement perfusion after medullary cavity reaming and pressurizing. Group IV (blank control group): The medullary cavity was not perfused with bone cement after reaming. In each group, the rabbits underwent femoral head resection and medullary cavity reaming. Before bone cement perfusion, 2 ml of developing tracer was injected through the ear vein. Radionuclide imaging was performed at 60, 120, and 180 minutes after bone cement perfusion, and the pulmonary radioactivity in vivo was measured. The rabbits were immediately sacrificed, and the pulmonary tissue was removed and its radioactivity was measured in vitro. Pulmonary tissue was then fixed and the pulmonary embolism and the associated pathological changes were observed.</p><p><b>RESULTS</b>The pulmonary radioactivity in vivo was measured at 60, 120, and 180 minutes after bone cement perfusion. The radioactivities of the four groups were 11.67 ± 2.16, 14.59 ± 2.92 and 18.43 ± 4.83 in group I; 8.37 ± 3.05, 10.35 ± 2.24 and 11.48 ± 2.96 in group II; 3.91 ± 1.19, 5.53 ± 2.95 and 7.25 ± 1.26 in group III; 1.04 ± 0.35, 1.14 ± 0.87 and 1.43 ± 0.97 in group IV. The radioactivities of groups I, II, III at 60, 120 and 180 minutes were significantly higher than group IV (P < 0.05). The pulmonary embolism could be detected. Pretreatment with epinephrine hydrochloride and fibrin sealant significantly decreased the pulmonary radioactivity in group II and group III, but it was still higher than in the group IV.</p><p><b>CONCLUSIONS</b>Radioimmunoimaging is an alternative method for the dynamic observation of rabbit pulmonary embolism after bone cement perfusion. Radioimmunoimaging is the optional way to evaluate the effect of pretreatment with epinephrine hydrochloride or fibrin sealant on pulmonary embolism after bone cement perfusion.</p>

Novel poly(ethylene-co-acrylic acid) nanofibrous biomaterials for peptide synthesis and biomedical applications.

Poly(ethylene-co-acrylic acid) (PE-co-AA) fibers in sizes of 200-500 nm were prepared by using a novel melt-extrusion-extraction fabrication process. The thermoplastic nanofibers could be controllably dispersed and reassembled by a novel solvent exchange filtration method. The dispersed PE-co-AA nanofibers possess active surface areas and could directly conduct chemical reactions on surfaces. Surface modifications and organic synthesis on the nanofibers were proven effective and controllable after the dispersion. Multistep synthesis of biomolecules, such as peptide ligand HWRGWV against Fc portion of human IgG, was successful. The surface-anchored ligand has shown bioactivity through selective binding to and staining by human IgG-alkaline phosphatase conjugate. Another peptide, LXY3, a selective cyclic peptide ligand against alpha3beta1 integrin of MDA-MB-231 breast cancer cells, was also prepared on the surfaces of the dispersed nanofibers. The results showed that MDA-MB-231 cells were able to specifically bind to and grow on surfaces of the nanofibers that were functionalized with LXY3.

Olomoucine inhibits cathepsin L nuclear translocation, activates autophagy and attenuates toxicity of 6-hydroxydopamine.

The finding of nuclear translocation of cathepsin L and its ability to process the CDP/Cux transcription factor uncovers an important role of cathepsin L in control of cell cycle progression. As the expression of certain cell cycle regulators is associated with nigral neuronal death, the present study was sought to investigate if nuclear translocation of cathepsin L and expression of certain cyclins were induced in DA neurons by 6-hydroxydopamine (6-OHDA). The neuroprotective effects of the cell cycle inhibitor olomoucine against 6-OHDA-induced death of nigral neurons were examined. Using immunocytochemistry and real-time PCR we demonstrated that cyclin D1, cyclin B1 and proliferating cell nuclear antigen (PCNA) were aberrantly expressed in some dopaminergic neurons after 6-OHDA infusion. The nuclear translocation of cathepsin L and up-regulation of LC3, a protein involved in autophagy, were observed in nigral DA neurons. Olomoucine, a cyclin dependent kinase (CDK) inhibitor, reduced contralateral rotations and the loss of TH-positive neurons in substantia nigra induced by lesion with 6-OHDA. Pretreatment of rats or primary DA neurons with olomoucine resulted in a partial blockade of nuclear translocation of cathepsin L. Olomoucine also increased the expression of punctate LC3 immunoreactivity, indicating activation of autophagy. These findings suggest that olomoucine may exert neuroprotective effects through inhibiting cathepsin L nuclear translocation and activating autophagy.