In vitro protoscolicidal effects of lithocholic acid on protoscoleces of Echinococcus granulosus and its mechanism.

Surgery has been found to be the best choice of treatment for hydatidosis. However, leakage of cyst contents during surgery is the foremost reason for recurrence of hydatidosis. In this study, we investigated the in vitro efficacy of lithocholic acid (LCA) against Echinococcus granulosus protoscoleces. The protoscoleces were divided into a control group, an albendazole (ABZ) positive control group and LCA intervention groups at concentrations of 0.5, 1, 2, and 3 mmol/L and stained with 0.1% eosin for observation using an inverted microscope; the protoscolecal ultrastructure was examined with SEM and TEM; the activities of ROS, SOD, and caspase-3 were investigated using an ROS kit, SOD kit, and caspase-3 kit, respectively; the contents of HO-1 and NQO-1 were analyzed by enzyme-linked immunosorbent assay; and the expression level of cytochrome c (Ctyc) was analyzed by western blotting. Results: As the concentration of LCA increased, the survival rate of protoscoleces gradually decreased. The microstructure shows that the external shape and internal structure were gradually deformed and collapse. SOD, GSH, HO-1 and NQO-1 decreased more significantly in the 3 mmol/L LCA group. However, ROS levels gradually increased. LCA treatment for 3 days at all concentrations significantly increased caspase-3 activity and expression in a dose-dependent manner. LCA decreased the level of Ctyc protein in vitro. LCA demonstrated a parasiticidal effect on the protoscoleces of Echinococcus granulosus in vitro. LCA may induce apoptosis of E. granulosus protoscoleces by oxidative stress and mitochondrial pathways.

In vivo stress granule misprocessing evidenced in a FUS knock-in ALS mouse model.

Many RNA-binding proteins, including TDP-43, FUS, and TIA1, are stress granule components, dysfunction of which causes amyotrophic lateral sclerosis (ALS). However, whether a mutant RNA-binding protein disrupts stress granule processing in vivo in pathogenesis is unknown. Here we establish a FUS ALS mutation, p.R521C, knock-in mouse model that carries impaired motor ability and late-onset motor neuron loss. In disease-susceptible neurons, stress induces mislocalization of mutant FUS into stress granules and upregulation of ubiquitin, two hallmarks of disease pathology. Additionally, stress aggravates motor performance decline in the mutant mouse. By using two-photon imaging in TIA1-EGFP transduced animals, we document more intensely TIA1-EGFP-positive granules formed hours but cleared weeks after stress challenge in neurons in the mutant cortex. Moreover, neurons with severe granule misprocessing die days after stress challenge. Therefore, we argue that stress granule misprocessing is pathogenic in ALS, and the model we provide here is sound for further disease mechanistic study.

Dihydroartemisinin induces ER stress-dependent apoptosis of Echinococcus protoscoleces in vitro.

In this study, we investigated the effect of dihydroartemisinin on Echinococcus protoscoleces and explored the role of endoplasmic reticulum stress in this process. Echinococcus protoscoleces were collected and cultured in RPMI 1640 medium. Changes in the expressions of glucose-regulated protein 78 (GRP-78), caspase-12, and C/EBP homologous protein (CHOP) were assessed through confocal immunofluorescence and western blot analysis. Cell viability and morphological changes were observed under a light microscope. The ultrastructure of protoscoleces was observed by scanning electron microscopy and transmission electron microscopy. Caspase-3 activity was detected using an enzyme assay kit. After dihydroartemisinin treatment, the protoscoleces showed loss of viability, and morphological changes including soma contraction, blebs formation, hooks loss, microtrichia destruction, and development of lipid droplets was observed. The levels of caspase-12 and CHOP were increased within 2 days of dihydroartemisinin treatment. However, the levels of GRP-78, caspase-12, and CHOP were decreased in 4 days. Furthermore, caspase-3 activity was increased after treatment with different concentrations of dihydroartemisinin. Dihydroartemisinin can induce apoptosis in protoscoleces via the ER stress-caspase-3 apoptotic pathway in vitro. These results indicate that dihydroartemisinin is a potentially valuable therapeutic agent against echinococcosis.

Sodium arsenite augments sensitivity of Echinococcus granulosus protoscoleces to albendazole.

This study aimed to observe the effects of sodium arsenite (NaAsO2) on apoptosis of Echinococcus granulosus protoscoleces induced by albendazole (ABZ), and to explore the potential mechanism of NaAsO2. According to the following final concentrations, the experimental groups were divided into 10 µM NaAsO2, 20 µM NaAsO2, 80 µM ABZ, 10 µM NaAsO2+80 µM ABZ, and 20 µM NaAsO2+80 µM ABZ. Viability was detected with 0.1% eosin staining. The ultrastructural alterations were visualized by scanning electron microscopy. Caspase-3 activity was assessed with colorimetric assay. Meanwhile, ELISA or WST were applied to detect the activities of antioxidases in NaAsO2 treatment groups. The maximum protoscolicidal effect was seen with the combination 20 µM NaAsO2+80 µM ABZ. The ultrastructural damage detected after NaAsO2+ABZ incubation were greater than those caused by ABZ alone and its primary damage site was the tegument of the parasite. The caspase-3 activity was clearly higher in protoscoleces treated with the combination of NaAsO2+ABZ than when drugs were used separately. The activities of NQO-1, HO-1, GST, and SOD were significantly lower in protoscoleces incubated with NaAsO2 than the untreated controls (P < 0.05). According to our results, ABZ could induce protoscoleces apoptosis, and NaAsO2 could significantly augment sensitivity of protoscoleces to ABZ.

Toxic effects of arsenic trioxide on Echinococcus granulosus protoscoleces through ROS production, and Ca2+-ER stress-dependent apoptosis.

Cystic echinococcosis is a severe parasitic disease that commonly affects the liver and causes abscesses or rupture into the surrounding tissues, leading to multiple complications, such as shock, severe abdominal pain, and post-treatment abscess recurrence. Currently, there are no efficient measures to prevent these complications. We previously confirmed that arsenic trioxide (As2O3) exhibited in vitro cytotoxicity against Echinococcus granulosus protoscoleces. In the present study, we aimed to explore the mechanism of As2O3-induced E. granulosus protoscoleces apoptosis. After exposing E. granulosus protoscoleces to 0, 4, 6, and 8 µM As2O3, reactive oxygen species (ROS) level was detected by fluorescence microscopy; superoxide dismutase (SOD), and caspase-3 activities were measured; intracellular Ca2+ was detected by flow cytometry; GRP-78 and caspase-12 protein levels were measured by western blot analysis. Our results showed that the expression of caspase-3 was gradually increased and the expression of SOD was gradually decreased in As2O3-treated groups of protoscoleces. Simultaneously, fluorescence microscopy and flow cytometry showed that the ROS level and the intracellular Ca2+ level were increased in a time- and dose-dependent manner. Western blot analysis showed that the expressions of GRP-78 and caspase-12 were higher in As2O3-treated groups than in the control group. These results suggest that As2O3-induced apoptosis in E. granulosus protoscoleces is related to elevation of ROS level, disruption of intracellular Ca2+ homeostasis, and endoplasmic reticulum stress. These mechanisms can be targeted in the future by safer and more effective drugs to prevent recurrence of cystic echinococcosis.

A comparative study of magnetic resonance imaging on the gray matter and resting-state function in prodromal and first-episode schizophrenia.

It is very difficult to predict the future development possibility of schizophrenia through the clinical symptoms of the high-risk cases. Therefore, how to determine the possibility of developing into schizophrenia individuals before the onset of the diseases are particularly important. The study investigated cerebral gray matter volume differences and resting-state functional connections among patients with psychosis risk syndrome (PRS), patients with first-episode schizophrenic (FES), and healthy controls (HC), aiming to provide scientific clinical evidence for schizophrenia early identification and intervention. A total of 19 PRS patients, 18 FES patients, and 29 HC were recruited. Gray matter volume and amplitude of low-frequency fluctuation (fALFF) during resting-state functional studies were measured. Comparison of gray matter volumes showed that PRS and FES groups had common reduced gray matter volume in the right caudate. PRS and FES patients showed altered connectivity mainly in the semantic processing-related brain areas. fALFF analysis found that PRF and FES patients had significant differences in fALFF values of the brain region mainly located in the subcortical network, visual recognition network, and auditory network. In addition, PRF individuals had a higher fALFF value and a lower fALFF value in the anterior wedge of the cerebral network than the HC group. Gray matter volume loss between related brain areas might appear prior to illness onset. Similar fALFF values occurred in PRS and FES groups indicated that multiple brain regions of neuronal activity abnormalities and unconventional neural network mechanism have been existed in PRS patients.

Effects of trigonelline inhibition of the Nrf2 transcription factor in vitro on Echinococcus granulosus.

The aim of this study was to investigate the impact of trigonelline (TRG) on Echinococcus granulosus, and to explore the inhibition impact of nuclear factor erythroid-2-related factor 2 (Nrf2) signaling pathway on E. granulosus protoscoleces. Echinococcus granulosus protoscoleces were incubated with various concentrations of TRG, and then Nrf2 protein expression and its localization in protoscoleces were detected by western blot analysis and immunofluorescence assay, respectively. Reactive oxygen species (ROS) level in protoscoleces was measured using ROS detection kit. Caspase-3 activity was measured using a caspase-3 activity assay kit, and NAD(P)H quinone oxidoreductase (NQO)-1 and heme oxygenase (HO)-1 activities in protoscoleces were measured by ELISA. The effect of TRG on protoscoleces viability was investigated using 0.1% eosin staining, and ultrastructural alterations in protoscoleces were examined by scanning electron microscopy (SEM). Immunolocalization experiment clearly showed that Nrf2 protein was predominantly present in cells of protoscoleces. TRG treatment reduced NQO-1 and HO-1 activities in protoscoleces, but could increase ROS level at early time. Protoscoleces could not survive when treated with 250 µM TRG for 12 days. SEM results showed that TRG-treated protoscoleces presented damage in the protoscoleces region, including hook deformation, lesions, and digitiform protuberance. Nrf2 protein expression was significantly decreased and caspase-3 activity was clearly increased in protoscoleces treated with TRG for 24 and 48 h, respectively, when compared with that in controls (P < 0.05). Our results demonstrated that TRG had scolicidal activity against E. granulosus protoscoleces. Nrf2 protein was mainly expressed in the cells and TRG could efficiently inhibit the Nrf2 signaling pathway in E. granulosus.

Improved electromagnetic wave absorption of Co nanoparticles decorated carbon nanotubes derived from synergistic magnetic and dielectric losses.

In this study, we report a feasible way to synthesize carbon nanotube nanocomposites deposited with cobalt nanoparticles (20-30 nm) on the surface (Co/CNTs) to serve as an electromagnetic (EM) wave absorption material. EM absorption measurements indicated that epoxy resin composites with 20 wt% Co/CNTs exhibited an effective EM absorption (RL < -10 dB) in the frequency range of 2.5-20 GHz with an absorber thickness of 1.0 to 6.0 mm. A strong absorption peak (RL = -36.5 dB) appeared at 4.1 GHz as the thickness was 4.0 mm, and the absorption bandwidth (RL < -10 dB) was in the frequency range of 3.6-4.6 GHz. The electromagnetic loss research suggested that the superior EM absorption performances including a light weight, strong absorption, broad frequency scope, and thin thickness could be ascribed to the synergistic effect of magnetic loss from Co nanoparticles and the dielectric loss of CNTs, resulting in better impedance matching.

Protoscolicidal effects of chenodeoxycholic acid on protoscoleces of Echinococcus granulosus.

Dissemination of protoscoleces-rich fluid during surgical operation for cystic echinococcosis is a major cause of its recurrence. Instillation of a scolicidal agent into hydatid cysts to reduce the risk of spillage of viable protoscoleces is an integral part of the surgical technique employed by many surgeons. In this study, the protoscolicidal effect of chenodeoxycholic acid (CDCA) was investigated. Freshly isolated protoscoleces were subjected to CDCA treatment (500, 1000, 2000, and 3000 µmol/L), and the effects on protoscoleces were investigated with the help of 0.1% eosin staining, electron microscopy, and colorimetric assay of caspase-3 like activity. Dose-dependent mortality of Echinococcus granulosus protoscoleces was observed within a few days of CDCA treatment. The treated protoscoleces showed loss of viability, and morphological changes such as contraction of the soma region, formation of blebs, rostellar disorganization, loss of hooks, destruction of microtriches, and formation of vesicles, lipid droplets, and lamellar bodies. Apoptosis was evident in the treated protoscoleces, as compared to the control group, which were cultivated for nearly 3 months. Our study indicates a therapeutic potential for CDCA as a protoscolicidal agent against E. granulosus. However, further studies are needed to test the long-term effects of CDCA in animal models.

Arsenic trioxide negatively affects Echinococcus granulosus.

Spillage of cyst contents during surgery is the major cause of recurrences of hydatidosis, also called cystic echinococcosis (CE). Currently, many scolicidal agents are used for inactivation of the cyst contents. However, due to complications in the use of those agents, new and more-effective treatment options are urgently needed. The aim of this study was to investigate the in vitro efficacy of arsenic trioxide (ATO) against Echinococcus granulosus protoscolices. Protoscolices of E. granulosus were incubated in vitro with 2, 4, 6, and 8 µmol/liter ATO; viability of protoscolices was assessed daily by microscopic observation of movements and 0.1% eosin staining. A small sample from each culture was processed for scanning and transmission electron microscopy. ATO demonstrated a potent ability to kill protoscolices, suggesting that ATO may represent a new strategy in treating hydatid cyst echinococcosis. However, the in vivo efficacy and possible side effects of ATO need to be explored.

Effects of protoscoleces excretory-secretory products of Echinococcus granulosus on hepatocyte growth, function, and glucose metabolism.

Cystic echinococcosis (CE) is one of the most widespread and harmful zoonotic parasitic diseases, which most commonly affects the liver. In this study, we characterized multiple changes in mouse hepatocytes following treatment with excretory-secretory products (ESPs) of Echinococcus granulosus protoscoleces (Eg-PSCs) by a factorial experiment. The cell counting kit-8 assay (CCK-8), the 5-ethynyl-2'-deoxyuridine (EdU) assay, and flow cytometry were used to detect the growth of hepatocytes. Inverted microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) were used to observe the morphology and ultrastructure of hepatocytes. An automatic biochemical analyzer and an ELISA detection kit were used to determine six conventional hepatocyte enzymatic indices, the levels of five hepatocyte-synthesized substances, and the contents of glucose and lactate. Western blot analysis was conducted to analyze the protein expression of three apoptosis-related proteins, Bax, Bcl-2, cleaved caspase-3, and six glucose metabolism pathways rate-limiting enzymes in hepatocytes. The results showed that ESPs inhibited hepatocyte proliferation and promoted hepatocyte apoptosis. The cell membrane and microvilli of hepatocytes changed, and the nucleus, mitochondria and rough endoplasmic reticulum were damaged to varying degrees. The contents of iron, albumin (ALB), uric acid (UA) and urea were increased, and the activities of six enzymes in hepatocytes were increased except for the decrease of transferrin (TRF). The expression levels of all six key enzymes in the glucose metabolism pathway in hepatocytes were reduced. Our characterization provides a basis for further research on the pathogenesis, prevention and treatment of CE.

The relationship between obesity and neurocognitive function in Chinese patients with schizophrenia.

BACKGROUND: Studies have reported that up to 60% of individuals with schizophrenia are overweight or obese. This study explored the relationship between obesity and cognitive performance in Chinese patients with schizophrenia. METHODS: Outpatients with schizophrenia aged 18-50 years were recruited from 10 study sites across China. Demographic and clinical information was collected. A neuropsychological battery including tests of attention, processing speed, learning/memory, and executive functioning was used to assess cognitive function, and these 4 individual domains were transformed into a neurocognitive composite z score. In addition, height and weight were measured to calculate body mass index (BMI). Patients were categorized into 4 groups (underweight, normal weight, overweight and obese) based on BMI cutoff values for Asian populations recommended by the World Health Organization. RESULTS: A total number of 896 patients were enrolled into the study. Fifty-four percent of participants were overweight or obese. A higher BMI was significantly associated with lower scores on the Wechsler Memory Scale-Revised (WMS-R) Visual Reproduction subscale, the Wechsler Adult Intelligence Scale-Revised (WAIS-R) Digit Symbol subscale, and the composite z score (p's ≤ 0.024). Obese patients with schizophrenia had significantly lower scores than normal weight patients on the Trail Making Test B, the WMS-R Visual Reproduction subscale, the WAIS Digit Symbol subscale, and the composite z score (p's ≤ 0.004). CONCLUSIONS: Our study suggests that, in addition to its well established risk for various cardiometabolic conditions, obesity is also associated with decreased cognitive function in Chinese patients with schizophrenia. Future studies should explore if weight loss and management can improve cognitive function in obese patients who suffer from schizophrenia.

In vitro and in vivo treatments of Echinococcus granulosus with Huaier aqueous extract and albendazole liposome.

The aim of this study was to investigate the in vitro and in vivo efficacies of chemotherapy employing albendazole liposome (L-ABZ), Huaier aqueous extract, and a Huaier aqueous extract/L-ABZ combination against Echinococcus granulosus. Protoscolices of E. granulosus were incubated in vitro with the two drugs, either separately or in combination, at the following final concentrations: 2 mg/mL Huaier aqueous extract, 10 µg/mL L-ABZ, and 2 mg/mL Huaier aqueous extract + 10 µg/mL L-ABZ. Huaier aqueous extract and L-ABZ displayed slower protoscolicidal activity when applied separately than when used in combination. The maximum protoscolicidal effect was found with the combination Huaier aqueous extract + L-ABZ. Despite the low Huaier aqueous extract + L-ABZ concentrations used, protoscolex viability dropped rapidly. In vivo studies were performed on mice injected with protoscolices of E. granulosus. Huaier aqueous extract and L-ABZ were administered three times a week for a period of 4 months by the oral route. Huaier aqueous extract in E. granulosus-infected mice was effective. Combined application of both drugs did increase the treatment efficacy. In conclusion, the outcomes obtained clearly demonstrated that in vitro and in vivo treatment with Huaier aqueous extract and L-ABZ is effective against E. granulosus.

In vitro effects of SB202190 on Echinococcus granulosus.

Spillage of cyst contents during surgical operation is the major cause of recurrence after hydatid cyst surgery. Instillation of a scolicidal agent into a hepatic hydatid cyst is the most commonly employed measure to prevent this complication. SB202190 is a pyridinyl imidazole derivative and is known to be a specific inhibitor of p38 MAPK. In the present study, the scolicidal effect of SB202190 was investigated. Freshly isolated Echinococcus granulosus protoscolices were subjected to SB202190 treatment (10, 20, 40, and 80 µM), and the effects on parasite viability were monitored by trypan blue staining. Corresponding effects were visualized by scanning and transmission electron microscopy. Dose-dependent protoscolex death within a few days of SB202190 treatment was observed. Although the in vitro scolicidal effect of SB202190 was satisfactory, the in vivo efficacy of this drug and also possible side effects remain to be further investigated.

[Advances in research on the MAPK signal transduction pathway of Echinococcus].

Mitogen-activated protein kinase (MAPK) is an important signaling transduction molecules, which can enter the nucleus and activate target gene when it was stimulated and become phosphorylation. MAPK signaling pathway is closely associated with various diseases. Recent studies have indicated that MAPK signaling transduction pathway is also involved in the growth and development of Echinococcus. This review summarizes the progress on the relationship between MAPK signal pathway and Echinococcus.

Propofol Induces the Expression of Nrf2 and HO-1 in Echinococcus granulosus via the JNK and p38 Pathway In Vitro.

The purpose of this study was to establish the relationship between mitogen-activated protein kinase (MAPK) and Nrf2 signaling pathways in Echinococcus granulosus (E. granulosus). E. granulosus protoscoleces (PSCs) cultured in vitro were divided into different groups: a control group, PSCs were pretreated with various concentrations of propofol followed by exposure to hydrogen peroxide (H2O2), and PSCs were pretreated with MAPK inhibitors, then co-treated with propofol and incubated in the presence of H2O2. PSCs activity was observed under an inverted microscope and survival rate was calculated. Reactive oxygen species (ROS) was detected by fluorescence microscopy, western blotting was used to detect the expression of Nrf2, Bcl-2, and heme oxygenase 1 (HO-1) in the PSCs among different groups. Pretreatment of PSCs with 0-1 mM propofol for 8 h prevented PSCs death after exposure to 0.5 mM H2O2. PSCs were pretreated with PD98059, SB202190, or SP600125 for 2 h, co-treated with propofol for an additional 8 h, and then exposed to 0.5 mM H2O2 for 6 h. On day 6, the PSCs viability was 42% and 39% in the p38 and JNK inhibitor groups, respectively. Additionally, pretreatment with propofol significantly attenuated the generation of ROS following H2O2 treatment. Propofol increased the expression of Nrf2, HO-1, and BCL2 compared with that of the control group. Pretreatment PSCs with SP600125 or SB202190, co-incubation with propofol and H2O2, can reduce the expression of Nrf2, HO-1, and BCL2 (p < 0.05). These results suggest that propofol induces an upregulated expression of HO-1 and Nrf2 by activation of the JNK and p38 MAPK signaling pathways. This study highlights the cross role of metabolic regulation of ROS signaling and targeting signalling pathways that may provide a promising strategy for the treatment of E. granulosus disease.

Hsa_circ_0128846 knockdown attenuates the progression of pancreatic cancer by targeting miR-1270/NR3C1 axis.

The considerable role of circular RNAs (circRNAs) make them prospective biomarkers in cancer therapy. Our study aimed to unveil the function of circ_0128846 in pancreatic cancer (PC). The expressions of circ_0128846, miR-1270 and NR3C1 mRNA were measured via RT-qPCR. The expressions of NR3C1 protein and apoptosis-related markers (Bax and Bcl-2) were measured via western blotting. CCK-8, colony-forming, or wound healing assay was respectively utilized to identify cell proliferation, growth and migration. Xenograft model was developed to evaluate tumor growth affected by circ_0128846 in vivo. The putative binding between miR-1270 and circ_0128846 or NR3C1 was testified by dual-luciferase reporter, RIP or pull-down assay. Circ_0128846 showed elevated expression in PC. Circ_0128846 deficiency restrained cancer cell proliferation, colony formation and migratory ability, enhanced cell apoptotic rate, and also impeded tumor development in vivo. Circ_0128846 directly targeted miR-1270 whose expression was declined in PC. The suppressive effects of silencing circ_0128846 on PC cell malignant phenotypes were largely reversed by miR-1270 inhibition. NR3C1 was targeted by miR-1270 and was highly regulated in PC. The repressive effects of NR3C1 knockdown on PC cell malignant phenotypes were partly abolished by miR-1270 inhibition. Circ_0128846 deficiency blocked PC progression via mediating the miR-1270/NR3C1 pathway, which partly illustrated PC pathogenesis.

Treatment of complicated hepatic cystic hydatidosis with intrabiliary rupture by pericystectomy in combination with Roux-en-Y hepaticojejunostomy.

This study retrospectively reviewed 9 cases of complicated hepatic cystic hydatidosis with intrabiliary rupture who were surgically treated with pericystectomy in combination with Roux-en-Y hepaticojejunostomy in our hospital from 2004 to 2010. The clinical features, results of laboratory tests, B-mode ultrasonography and CT, post-operative recovery, days of hospital stay after the operation and post-operative complications were statistically analyzed and the patients were followed up. The subjects in our series included 7 males and 2 females, whose average age was 50.78±7.58 years. Before operation, 9 patients suffered from pain of the right upper quadrant and jaundice, which, in 4 cases (44.45%), were accompanied with fever and chills. Preoperative B-mode ultrosonography and CT showed that all the 9 patients had single hydatid cyst, with their diameter being 9.33±1.58 cm on average. The lesions involved segments V, VI in 6 cases, and segment IV in 3 cases. By WHO classification, 7 cases were classified as CE3 and 2 cases as CE4. They all had choledochectasia. The subjects underwent the surgery uneventfully. Intraoperatively, 2-4 biliary fistula orifices were found, with the average of the orifice being (0.79±0.20) cm. After the operation, one patient developed incision infection, one had pulmonary infection and one suffered from reflux cholangitis. No anastomotic leaks or peri-operative deaths took place and follow-up revealed no recurrence and implantative metastasis. We are led to conclude that pericystectomy in combination with Roux-en-Y hepaticojejunostomy can achieve satisfactory results for the treatment of complicated hepatic cystic hydatidosis with intrabiliary rupture.

Dual-gRNA approach with limited off-target effect corrects C9ORF72 repeat expansion in vivo.

C9ORF72 GGGGCC repeat expansion is the most common genetic cause for amyotrophic lateral sclerosis and frontotemporal dementia, which generates abnormal DNA and RNA structures and produces toxic proteins. Recently, efficacy of CRISPR/Cas9-mediated editing has been proven in treatment of disease. However, DNA low complexity surrounding C9ORF72 expansion increases the off-target risks. Here we provide a dual-gRNA design outside of the low complexity region which enables us to remove the repeat DNA in a 'cutting-deletion-fusion' manner with a high fusion efficiency (50%). Our dual-gRNA design limits off-target effect and does not significantly affect C9ORF72 expression. In neurons carrying patient C9ORF72 expansion, our approach removes the repeat DNA and corrects the RNA foci in vitro and in vivo. Therefore, we conclude that our proof-of-concept design correct C9ORF72 repeat expansion, which may have potential therapeutic value for the patients.