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1.
Acta Pharmaceutica Sinica B ; (6): 781-794, 2024.
Article in English | WPRIM | ID: wpr-1011269

ABSTRACT

Small interfering RNA (siRNA) has a promising future in the treatment of ocular diseases due to its high efficiency, specificity, and low toxicity in inhibiting the expression of target genes and proteins. However, due to the unique anatomical structure of the eye and various barriers, delivering nucleic acids to the retina remains a significant challenge. In this study, we rationally design PACD, an A-B-C type non-viral vector copolymer composed of a hydrophilic PEG block (A), a siRNA binding block (B) and a pH-responsive block (C). PACDs can self-assemble into nanosized polymeric micelles that compact siRNAs into polyplexes through simple mixing. By evaluating its pH-responsive activity, gene silencing efficiency in retinal cells, intraocular distribution, and anti-angiogenesis therapy in a mouse model of hypoxia-induced angiogenesis, we demonstrate the efficiency and safety of PACD in delivering siRNA in the retina. We are surprised to discover that, the PACD/siRNA polyplexes exhibit remarkable intracellular endosomal escape efficiency, excellent gene silencing, and inhibit retinal angiogenesis. Our study provides design guidance for developing efficient nonviral ocular nucleic acid delivery systems.

2.
China Pharmacy ; (12): 1584-1589, 2023.
Article in Chinese | WPRIM | ID: wpr-977846

ABSTRACT

OBJECTIVE To explore the intervention effect and related mechanism of Tongxinluo capsule on renal fibrosis in rats with diabetic nephropathy (DN). METHODS Eight rats were selected as control group (ordinary feed), the remaining rats were given high-glucose and high-fat diet combined with ip injection of streptozotocin (35 mg/kg) to induce DN model. Model rats were randomly divided into model group (purified water), irbesartan group (positive control, 14.12 mg/kg) and Tongxinluo capsule group (0.3 g/kg), including 12 rats in the model group and 11 rats for each of the other two groups. All groups were given relevant medicine or water intragastrically, once a day, for 16 consecutive weeks. After the last medication, fasting blood glucose and 24 h urinary total protein (24 h UTP) were detected. Pathological changes in renal cortex of rats in each group were observed. Serum levels of tissue-type plasminogen activator (PA) and plasminogen activator inhibitor 1 (PAI-1) were measured. mRNA expressions of transforming growth factor-β(1 TGF-β1), type Ⅳ collagen(COL-Ⅳ), Wnt4 and β-catenin in renal cortex of rats were detected. The protein depositions or expressions of TGF-β1, COL-Ⅳ, focal adhesion kinase (FAK), integrin-linked kinase (ILK), E-cadherin, PA, PAI-1, Wnt4 and β-catenin in renal cortex of rats were observed or determined. RESULTS Compared with model group, 24 h UTP of rats in Tongxinluo capsule group were all significantly reduced (P<0.05); pathological damage and fibrosis of renal cortex were relieved; the expression of PA in serum and renal cortex was significantly increased, while PAI-1 level was significantly reduced (P<0.05); the depositions of COL-Ⅳ and TGF-β1 in renal cortex were all reduced, and corresponding mRNA expression was decreased significantly (P<0.05); the depositions of ILK and FAK were decreased, while the deposition of E-cadherin was increased; protein and mRNA expressions of Wnt4 and β-catenin were significantly reduced (P<0.05). CONCLUSIONS Tongxinluo capsule can relieve pathological damage to renal tissue and renal fibrosis of DN model rats, and reduce extracellular matrix deposition. The mechanism may be related to regulation of fibrinolytic system activity, the decrease of ILK and FAK expression, and inhibition of Wnt/β-catenin signaling pathway.

3.
Journal of Geriatric Cardiology ; (12): 577-585, 2023.
Article in English | WPRIM | ID: wpr-1010185

ABSTRACT

OBJECTIVE@#To develop and validate a user-friendly risk score for older mitral regurgitation (MR) patients, referred to as the Elder-MR score.@*METHODS@#The China Senile Valvular Heart Disease (China-DVD) Cohort Study functioned as the development cohort, while the China Valvular Heart Disease (China-VHD) Study was employed for external validation. We included patients aged 60 years and above receiving medical treatment for moderate or severe MR (2274 patients in the development cohort and 1929 patients in the validation cohort). Candidate predictors were chosen using Cox's proportional hazards model and stepwise selection with Akaike's information criterion.@*RESULTS@#Eight predictors were identified: age ≥ 75 years, body mass index < 20 kg/m2, NYHA class III/IV, secondary MR, anemia, estimated glomerular filtration rate < 60 mL/min per 1.73 m2, albumin < 35 g/L, and left ventricular ejection fraction < 60%. The model displayed satisfactory performance in predicting one-year mortality in both the development cohort (C-statistic = 0.73, 95% CI: 0.69-0.77, Brier score = 0.06) and the validation cohort (C-statistic = 0.73, 95% CI: 0.68-0.78, Brier score = 0.06). The Elder-MR score ranges from 0 to 15 points. At a one-year follow-up, each point increase in the Elder-MR score represents a 1.27-fold risk of death (HR = 1.27, 95% CI: 1.21-1.34, P < 0.001) in the development cohort and a 1.24-fold risk of death (HR = 1.24, 95% CI: 1.17-1.30, P < 0.001) in the validation cohort. Compared to EuroSCORE II, the Elder-MR score demonstrated superior predictive accuracy for one-year mortality in the validation cohort (C-statistic = 0.71 vs. 0.70, net reclassification improvement = 0.320, P < 0.01; integrated discrimination improvement = 0.029, P < 0.01).@*CONCLUSIONS@#The Elder-MR score may serve as an effective risk stratification tool to assist clinical decision-making in older MR patients.

4.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 1104-1109, 2023.
Article in Chinese | WPRIM | ID: wpr-998236

ABSTRACT

ObjectiveTo develop a bilateral rehabilitation robot motion assistance strategy based on admittance control, so that rehabilitation physicians can assist patients in rehabilitation training through remote teaching. MethodsA bilateral remote rehabilitation platform with upper limb terminal traction was constructed. Based on the velocity admittance control, the interactive movement between the master robot and the rehabilitation physician was realized, and the position information transmission of the master-slave robot was realized through the communication framework built. The slave robot received the position coordinates of the main robot, and drove the patient to carry out rehabilitation exercises under the attitude admittance controller. ResultsThe robot could drive the patient to accurately track the trajectory of the doctor's teaching in real time, and improve the safety and compliance of the training and human-computer interaction. ConclusionBy introducing two admittance controllers, the trajectory of the physician's end can be accurately tracked when driving the patient's movement from the robotic arm, which effectively avoids the discomfort of the patient's arm in process of rehabilitation.

5.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 302-311, 2023.
Article in Chinese | WPRIM | ID: wpr-971872

ABSTRACT

ObjectiveTo review the development history of research on people with disabilities, summarize the patterns, characteristics and deficiencies in this discipline, and make suggestions for discipline development in the future. MethodsThe literature about disability from 1986 to 2018 were searched and retrieved on the CNKI. Valid literature were selected based on the title and abstract. Descriptive analyses were used to analyze the development of research on people with disabilities in China. VOSviewer was used to explore the cooperation among researchers and research hotspots in this field. ResultsA total of 2 267 papers were included. Researches on people with disabilities in China started in 1986 and then experienced rapid development driven by survey data, showing obvious stage characteristics. The foundation of academic cooperation networks has been formed initially, showing the comprehensive development of multiple themes. However, in the new stage, the lack of follow-up support for research infrastructure conditions, as well as the slow innovation of research theories and expansion of research contents may become key factors hindering the further development of the discipline. ConclusionThe research foundation should be consolidated in the future, including broadening cooperation and communication channels, strengthening disability statistics, and promoting cross-disciplinary research. Theoretical research should be strengthened by standardizing research methods and finding internalized theoretical innovation points combining the national conditions. Finally, research content should be enriched, especially by closely combining the current changes in the needs of people with disabilities and strengthening the research on disability prevention and control, health promotion, social integration, and social management of the people with disabilities.

6.
China Journal of Chinese Materia Medica ; (24): 3565-3575, 2023.
Article in Chinese | WPRIM | ID: wpr-981488

ABSTRACT

This study aimed to investigate the underlying mechanism of Zhenwu Decoction in the treatment of heart failure by regulating electrical remodeling through the transient outward potassium current(I_(to))/voltage-gated potassium(Kv) channels. Five normal SD rats were intragastrically administered with Zhenwu Decoction granules to prepare drug-containing serum, and another seven normal SD rats received an equal amount of distilled water to prepare blank serum. H9c2 cardiomyocytes underwent conventional passage and were treated with angiotensin Ⅱ(AngⅡ) for 24 h. Subsequently, 2%, 4%, and 8% drug-containing serum, simvastatin(SIM), and BaCl_2 were used to interfere in H9c2 cardiomyocytes for 24 h. The cells were divided into a control group [N, 10% blank serum + 90% high-glucose DMEM(DMEM-H)], a model group(M, AngⅡ + 10% blank serum + 90% DMEM-H), a low-dose Zhenwu Decoction-containing serum group(Z1, AngⅡ + 2% drug-containing serum of Zhenwu Decoction + 8% blank serum + 90% DMEM-H), a medium-dose Zhenwu Decoction-containing serum group(Z2, AngⅡ + 4% drug-containing serum of Zhenwu Decoc-tion + 6% blank serum + 90% DMEM-H), a high-dose Zhenwu Decoction-containing serum group(Z3, AngⅡ + 8% drug-containing serum of Zhenwu Decoction + 2% blank serum + 90% DMEM-H), an inducer group(YD, AngⅡ + SIM + 10% blank serum + 90% DMEM-H), and an inhibitor group(YZ, AngⅡ + BaCl_2 + 10% blank serum + 90% DMEM-H). The content of ANP in cell extracts of each group was detected by ELISA. The relative mRNA expression levels of ANP, Kv1.4, Kv4.2, Kv4.3, DPP6, and KChIP2 were detected by real-time quantitative PCR. The protein expression of Kv1.4, Kv4.2, Kv4.3, DPP6, and KChIP2 was detected by Western blot. I_(to) was detected by the whole cell patch-clamp technique. The results showed that Zhenwu Decoction at low, medium, and high doses could effectively reduce the surface area of cardiomyocytes. Compared with the M group, the Z1, Z2, Z3, and YD groups showed decreased ANP content and mRNA level, increased protein and mRNA expression of Kv4.2, Kv4.3, DPP6, and KChIP2, and decreased protein and mRNA expression of Kv1.4, and the aforementioned changes were the most notable in the Z3 group. Compared with the N group, the Z1, Z2, and Z3 groups showed significantly increased peak current and current density of I_(to). The results indicate that Zhenwu Decoction can regulate myocardial remodeling and electrical remodeling by improving the expression trend of Kv1.4, Kv4.2, Kv4.3, KChIP2, and DPP6 proteins and inducing I_(to) to regulate Kv channels, which may be one of the mechanisms of Zhenwu Decoction in treating heart failure and related arrhythmias.


Subject(s)
Rats , Animals , Myocytes, Cardiac , Atrial Remodeling , Rats, Sprague-Dawley , Heart Failure/metabolism , RNA, Messenger/metabolism , Potassium
7.
Chinese Medical Sciences Journal ; (4): 191-205, 2023.
Article in English | WPRIM | ID: wpr-1008987

ABSTRACT

Background Kidney renal clear cell carcinoma (KIRC) is one of the most common renal malignancies with a high mortality rate. Cuproptosis, a novel form of cell death, is strongly linked to mitochondrial metabolism and is mediated by protein lipoylation, leading to a proteotoxic stress response and cell death. To date, few studies have ellucidated the holistic role of cuproptosis-related genes (CRGs) in the pathogenesis of KIRC.Methods We comprehensively and completely analyzed the RNA sequencing data and corresponding clinical information from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. We screened for differentially expressed CRGs and constructed a prognostic risk model using univariate and multivariate Cox proportional regression analyses. Kaplan-Meier analysis was performed and receiver operating characteristic (ROC) curves were plotted to predict the prognosis of KIRC patients. Functional enrichment analysis was utilized to explore the internal mechanisms. Immune-related functions were analyzed using single-sample gene set enrichment analysis (ssGSEA), tumour immune dysfunction and exclusion (TIDE) scores, and drug sensitivity analysis.Results We established a concise prognostic risk model consisting of four CRGs (DBT, DLAT, LIAS and PDHB) to predict the overall survival (OS) in KIRC patients. The results of the survival analysis indicated a significantly lower OS in the high-risk group as compared to the patients in the low-risk group. The area under the time-dependent ROC curve (AUC) at 1, 3, and 5 year was 0.691, 0.618, and 0.614 in KIRC. Functional enrichment analysis demonstrated that CRGs were significantly enriched in tricarboxylic acid (TCA) cycle-related processes and metabolism-related pathways. Sorafenib, doxorubicin, embelin, and vinorelbine were more sensitive in the high-risk group.Conclusions We constructed a concise CRGs risk model to evaluate the prognosis of KIRC patients and this may be a new direction for the diagnosis and treatment of KIRC.


Subject(s)
Humans , Carcinoma, Renal Cell/genetics , Immunotherapy , Kidney , Kidney Neoplasms/genetics , Prognosis , Copper , Apoptosis
8.
Acta Pharmaceutica Sinica B ; (6): 2234-2249, 2023.
Article in English | WPRIM | ID: wpr-982848

ABSTRACT

The many-banded krait, Bungarus multicinctus, has been recorded as the animal resource of JinQianBaiHuaShe in the Chinese Pharmacopoeia. Characterization of its venoms classified chief phyla of modern animal neurotoxins. However, the evolutionary origin and diversification of its neurotoxins as well as biosynthesis of its active compounds remain largely unknown due to the lack of its high-quality genome. Here, we present the 1.58 Gbp genome of B. multicinctus assembled into 18 chromosomes with contig/scaffold N50 of 7.53 Mbp/149.8 Mbp. Major bungarotoxin-coding genes were clustered within genome by family and found to be associated with ancient local duplications. The truncation of glycosylphosphatidylinositol anchor in the 3'-terminal of a LY6E paralog released modern three-finger toxins (3FTxs) from membrane tethering before the Colubroidea divergence. Subsequent expansion and mutations diversified and recruited these 3FTxs. After the cobra/krait divergence, the modern unit-B of β-bungarotoxin emerged with an extra cysteine residue. A subsequent point substitution in unit-A enabled the β-bungarotoxin covalent linkage. The B. multicinctus gene expression, chromatin topological organization, and histone modification characteristics were featured by transcriptome, proteome, chromatin conformation capture sequencing, and ChIP-seq. The results highlighted that venom production was under a sophisticated regulation. Our findings provide new insights into snake neurotoxin research, meanwhile will facilitate antivenom development, toxin-driven drug discovery and the quality control of JinQianBaiHuaShe.

9.
Chinese Journal of Schistosomiasis Control ; (6): 52-58, 2022.
Article in Chinese | WPRIM | ID: wpr-920744

ABSTRACT

Objective To identify the differentially expressed proteins in different liver tissues in the mouse model of alveolar echinococcosis using high-resolution mass spectrometry with data independent acquisition (DIA), and to identify the key proteins contributing to the pathogenesis of alveolar echinococcosis. Methods Protoscoleces were isolated from Microtus fuscus with alveolar echinococcosis and the experimental model of alveolar echinococcosis was established in female Kunming mice aged 6 to 8 weeks by infection with Echinococcus multilocularis protoscoleces. Mice were divided into the experimental and control groups, and animals in the experimental group was injected with approximately 3 000 protoscoleces, while mice in the control group were injected with the same volume of physiological saline. Mouse liver specimens were sampled from both groups one year post-infection and subjected to pathological examinations. In addition, the lesions (the lesion group) and peri-lesion specimens (the peri-lesion group) were sampled from the liver of mice in the experimental group and the normal liver specimens (the normal group) were sampled from mice in the control group for DIA proteomics analysis, and the differentially expressed proteins were subjected to bioinformatics analysis. Results A total of 1 020 differentially expressed proteins were identified between the lesion group and the normal group, including 671 up-regulated proteins and 349 down-regulated proteins, and 495 differentially expressed proteins were identified between the peri-lesion group and the normal group, including 327 up-regulated proteins and 168 down-regulated proteins. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that these differentially expressed proteins were involved in peroxisome, peroxisome proliferator-activated receptor (PPAR) and fatty acid degradation pathways, and the peroxisome and PPAR signaling pathways were found to correlate with liver injury. Several differentially expressed proteins that may contribute to the pathogenesis of alveolar echinococcosis were identified in these two pathways, including fatty acid binding protein 1 (Fabp1), Acyl-CoA synthetase long chain family member 1 (Acsl1), Acyl-CoA oxidase 1 (Acox1), Enoyl-CoA hydratase and 3-hydroxyacyl CoA dehydrogenase (Ehhadh) and Acetyl-Coenzyme A acyltransferase 1B (Acaa1b), which were down-regulated in mice in the experimental group. Conclusion A large number of differentially expressed proteins are identified in the liver of the mouse model of alveolar echinococcosis, and Fabp1, Acsl1, Acox1, Ehhadh and Acaa1b may contribute to the pathogenesis of alveolar echinococcosis.

10.
Chinese Journal of Schistosomiasis Control ; (6): 41-51, 2022.
Article in Chinese | WPRIM | ID: wpr-920743

ABSTRACT

Objective To identify the differentially expressed proteins in different liver tissues in the mouse model of cystic echinococcosis (CE), so as to provide insights into the research and development of therapeutic drugs targeting CE. Methods Female Kunming mice at ages of 6 to 8 weeks were randomly assigned into the CE group and the control group. Mice in the CE group were intraperitoneally infected with 2 000 Echinococcus multilocularis protoscoleces, while mice in the control group were injected with the same volume of physiological saline. All mice in both groups were sacrificed after breeding for 350 d, and the lesions (the lesion group) and peri-lesion specimens (the peri-lesion group) were sampled from the liver of mice in the CE group and the normal liver specimens (the normal group) were sampled from mice in the control group for data independent acquisition (DIA) proteomics analysis, and the differentially expressed proteins were subjected to Gene Ontology (GO) term enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Results A total of 26 differentially expressed proteins were identified between the lesion group and the normal group and between the peri-lesion group and the normal group, including 8 up-regulated proteins and 18 down-regulated proteins. GO term enrichment analysis showed that these differentially expressed proteins were predominantly enriched in endoplasmic reticulum membrane (biological components), oxidoreductase activity (molecular function) and oxoacid metabolic process and monocarboxylic acid metabolic process (biological processes). KEGG pathway enrichment analysis revealed that the differentially expressed protein Acyl-CoA oxidase 1 (Acox1), which contributed to primary bile acid biosynthesis during the fatty acid oxidation, was involved in peroxisome signaling pathway, and the differentially expressed protein fatty acid binding protein 1 (Fabp1), which contributed to fatty acid transport, was involved in the peroxisome proliferator-activated receptor (PPAR) signaling pathway. Conclusion Differentially expressed proteins are identified in the liver specimens between mouse models of CE and normal mice, and some differentially expressed proteins may serve as potential drug targets for CE.

11.
Chinese Journal of Medical Instrumentation ; (6): 137-140, 2022.
Article in Chinese | WPRIM | ID: wpr-928874

ABSTRACT

Spatiotemporal gait parameters provide important information for the rehabilitation of patients with gait dysfunction. These parameters are often obtained by complex systems such as optical motioncapture system and pressure plates. However, these systems cannot be deployed at the lower-limb rehabilitation robot easily because of high costs, large area occupation and wearable requirements. We present a gait measurement system with a Light Detection And Ranging(LIDAR) laser sensor based on the lower-limb rehabilitation robot. Firstly, to calculate gait parameters, the data are aggregated into left and right legs by the clustering algorithm and the legs contour is fitted with two circles respectively according to the least square method. Then, the spatiotemporal gait parameters are defined based on the time and position of initial contact(IC) and toe off(TO). Finally, to verify the validity of the proposed system, we compared the results of the proposed system with a 3D motion capture system based on a lower-limb rehabilitation robot. Experimental results showed that the gait detection system can measure the parameters within a small range of error that testified the validation of the proposed system. This system proved to be a valid and reliable method for the measurement of gait parameters.


Subject(s)
Humans , Biomechanical Phenomena , Gait , Lasers , Lower Extremity , Motion , Robotics
12.
Chinese Journal of Medical Instrumentation ; (6): 21-27, 2022.
Article in Chinese | WPRIM | ID: wpr-928851

ABSTRACT

This study evaluates the static balance ability of human body based on lower limb rehabilitation robot.According to the balance parameters obtained from the movement trajectory of the center of human pelvis, SPSS statistical software was used to verify that there was significant difference between the two groups (P<0.01). Principal component analysis is used to allocate the weight of each parameter and establish the comprehensive evaluation value. The comprehensive evaluation value of the control group was 0.383±0.038, and the experimental group was 0.875±0.136. When the subject's comprehensive evaluation value is between 0.739 and 1.011, it indicates the presence of balance dysfunction, and when it is between 0.345~0.421, it indicates that the balance of the lower limbs of the subject is normal. Experimental results show that this evaluation method can objectively and quantitatively reflect the static equilibrium state of human body.


Subject(s)
Humans , Lower Extremity , Movement , Pelvis , Postural Balance
13.
Chinese Journal of Neurology ; (12): 612-619, 2021.
Article in Chinese | WPRIM | ID: wpr-885472

ABSTRACT

Epilepsy is one of the most common diseases of the central nervous system, affecting tens of millions of people around the world. Most of clinically used antiepileptic drugs are based on ion mechanism to antagonize epileptic seizures, targeted to various ion channels or ion channel receptors. However, with the in-depth research on the pathogenesis of epilepsy, the non-ionic antiepileptic mechanism has increasingly become the key to the control of various intractable epilepsy, and the relevant drugs have gradually achieved clinical transformation. In this paper, non-ionic antiepileptic mechanisms are classified to clinical and preclinical types according to whether clinical transformation has been achieved. The application of non-ionic antiepileptic drugs in refractory epilepsy was mainly introduced, including everolimus, cannabidiol, fenfluramine, padsevonil, medium chain triglyceride modified ketogenic diet, and anakinra. Additionally, some preclinical non-ionic antiepileptic mechanisms such as prostaglandin, adenosine, metabolic glutamate receptor and mitochondrial mechanism are briefly introduced. The authors believe that the current stage of ionic antiepileptic drugs research has reached the bottleneck of transformation and it is difficult to achieve a major breakthrough in the mechanism, but there are broader research prospects in non-ionic antiepileptic mechanisms because a large number of them have not yet been clinically transformed. From a deeper perspective, some non-ionic antiepileptic mechanisms may have been involved in the fundamental mechanism of epileptogenesis, and they may be the prospect for the future treatment of refractory epilepsy.

14.
Journal of Biomedical Engineering ; (6): 129-135, 2020.
Article in Chinese | WPRIM | ID: wpr-788887

ABSTRACT

In order to stimulate the patients' active participation in the process of robot-assisted rehabilitation training of stroke patients, the rehabilitation robots should provide assistant torque to patients according to their rehabilitation needs. This paper proposed an assist-as-needed control strategy for wrist rehabilitation robots. Firstly, the ability evaluation rules were formulated and the patient's ability was evaluated according to the rules. Then the controller was designed. Based on the evaluation results, the controller can calculate the assistant torque needed by the patient to complete the rehabilitation training task and send commands to motor. Finally, the motor is controlled to output the commanded value, which assists the patient to complete the rehabilitation training task. The control strategy was implemented to the wrist function rehabilitation robot, which could achieve the training effect of assist-as-needed and could avoid the surge of assistance torque. In addition, therapists can adjust multiple parameters in the ability evaluation rules online to customize the difficulty of tasks for patients with different rehabilitation status. The method proposed in this paper does not rely on the information from force sensor, which reduces development costs and is easy to implement.

15.
Chinese Journal of Neurology ; (12): 96-102, 2020.
Article in Chinese | WPRIM | ID: wpr-799510

ABSTRACT

Objective@#To investigate the effect of glycogen synthase kinase 3β (GSK3β) on the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 (Aβ31-35) in HT22 cells.@*Methods@#HT22 mouse hippocampal cells were divided into control group, Aβ31-35 group and LiCl+Aβ31-35 group by random number table method in the present study. Cells were synchronized to G0/G1 phase by 1% serum starvation for 1 hour (circadian time 0 (CT0)). Cell viability was detected by the cell counting kit-8 assay. The mRNA expression of clock gene Bmal1 was examined by real-time PCR at different CT times. The expression of GSK3β and BMAL1 protein was detected by Western blotting.@*Results@#Compared with the control group, Aβ31-35 induced the decreased expression of Bmal1 mRNA; The expression of both Bmal1 mRNA and BMAL1 protein was decreased significantly at CT20 (Bmal1 mRNA: 0.38±0.06 vs 0.83±0.08, t=4.549, P=0.001; BMAL1 protein: 0.67±0.04 vs 1.00±0.04, t=5.943, P<0.001). In the Aβ31-35 group, GSK3β activity was increased and the ratio of phosphorylated GSK3βS9 to GSK3β was decreased compared to the control group (0.66±0.08 vs 1.02±0.14, t=2.217, P=0.025). Aβ31-35 decreased the viability of HT22 cells (71.85%±6.20% in the Aβ31-35 group vs 98.14%±2.68% in the control group, t=3.891, P=0.006), and the GSK3β inhibitor LiCl pretreatment effectively reversed the decline of the viability induced by Aβ31-35 (90.74%±5.74% in the LiCl+Aβ31-35 group vs 71.85%±6.20% in the Aβ31-35 group, t=3.412, P=0.010). LiCl (in the LiCl+Aβ31-35 group) increased the expression of Bmal1 mRNA and BMAL1 protein significantly at CT20 compared with the Aβ31-35 group (Bmal1 mRNA: 0.72±0.05 vs 0.38±0.06, t=4.378, P=0.001; BMAL1 protein: 0.90±0.04 vs 0.67±0.04, t=4.052, P=0.002).@*Conclusion@#Increased GSK3β activity involved in the decreased expression of Bmal1 induced by Aβ31-35 in HT22 cells.

16.
Chinese Journal of Neurology ; (12): 96-102, 2020.
Article in Chinese | WPRIM | ID: wpr-799509

ABSTRACT

Objective@#To investigate the effect of glycogen synthase kinase 3β (GSK3β) on the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 (Aβ31-35) in HT22 cells.@*Methods@#HT22 mouse hippocampal cells were divided into control group, Aβ31-35 group and LiCl+Aβ 31-35 group by random number table method in the present study. Cells were synchronized to G0/G1 phase by 1% serum starvation for 1 hour (circadian time 0 (CT0)). Cell viability was detected by the cell counting kit-8 assay. The mRNA expression of clock gene Bmal1 was examined by real-time PCR at different CT times. The expression of GSK3β and BMAL1 protein was detected by Western blotting.@*Results@#Compared with the control group, Aβ31-35 induced the decreased expression of Bmal1 mRNA; The expression of both Bmal1 mRNA and BMAL1 protein was decreased significantly at CT20 (Bmal1 mRNA: 0.38±0.06 vs 0.83±0.08, t=4.549, P=0.001; BMAL1 protein: 0.67±0.04 vs 1.00±0.04, t=5.943, P<0.001). In the Aβ31-35 group, GSK3β activity was increased and the ratio of phosphorylated GSK3βS9 to GSK3β was decreased compared to the control group (0.66±0.08 vs 1.02±0.14, t=2.217, P=0.025). Aβ31-35 decreased the viability of HT22 cells (71.85%±6.20% in the Aβ31-35 group vs 98.14%±2.68% in the control group, t=3.891, P=0.006), and the GSK3β inhibitor LiCl pretreatment effectively reversed the decline of the viability induced by Aβ31-35 (90.74%±5.74% in the LiCl+Aβ31-35 group vs 71.85%±6.20% in the Aβ31-35 group, t=3.412, P=0.010). LiCl (in the LiCl+Aβ31-35 group) increased the expression of Bmal1 mRNA and BMAL1 protein significantly at CT20 compared with the Aβ31-35 group (Bmal1 mRNA: 0.72±0.05 vs 0.38±0.06, t=4.378, P=0.001; BMAL1 protein: 0.90±0.04 vs 0.67±0.04, t=4.052, P=0.002).@*Conclusion@#Increased GSK3β activity involved in the decreased expression of Bmal1 induced by Aβ31-35 in HT22 cells.

17.
Chinese Journal of Neurology ; (12): 96-102, 2020.
Article in Chinese | WPRIM | ID: wpr-870771

ABSTRACT

Objective To investigate the effect of glycogen synthase kinase 3β (GSK3β) on the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 (Aβ31-35) in HT22 cells.Methods HT22 mouse hippocampal cells were divided into control group,Aβ31-35 group and LiCl+Aβ 31-35 group by random number table method in the present study.Cells were synchronized to G0/G1 phase by 1% serum starvation for 1 hour (circadian time 0 (CT0)).Cell viability was detected by the cell counting kit-8 assay.The mRNA expression of clock gene Bmal1 was examined by real-time PCR at different CT times.The expression of GSK3β and BMAL1 protein was detected by Western blotting.Results Compared with the control group,Aβ31-35 induced the decreased expression of Bmal1 mRNA;The expression of both Bmal1 mRNA and BMAL1 protein was decreased significantly at CT20 (Bmal1 mRNA:0.38±0.06 vs 0.83±0.08,t=4.549,P=0.001;BMAL1 protein:0.67±0.04 vs 1.00±0.04,t=5.943,P<0.001).In the Aβ31-35group,GSK3β activity was increased and the ratio of phosphorylated GSK3βS9 to GSK3β was decreased compared to the control group (0.66±0.08 vs 1.02±0.14,t=2.217,P=0.025).Aβ31-35 decreased the viability of HT22 cells (71.85%±6.20% in the Aβ31-35 group vs 98.14%±2.68% in the control group,t=3.891,P=0.006),and the GSK3β inhibitor LiC1 pretreatment effectively reversed the decline of the viability induced by Aβ31-35 (90.74%±5.74% in the LiCl+Aβ31-35 group vs 71.85%±6.20% in the Aβ31-35 group,t=3.412,P=0.010).LiCl (in the LiCl+Aβ31-35 group) increased the expression of Bmal1 mRNA and BMAL1 protein significantly at CT20 compared with the Aβ31-35 group (Bmal1 mRNA:0.72±0.05 vs 0.38±0.06,t=4.378,P=0.001;BMAL1 protein:0.90±0.04 vs 0.67±0.04,t=4.052,P=0.002).Conclusion Increased GSK3β activity involved in the decreased expression of Bmal 1 induced by Aβ31-35 in HT22 cells.

18.
Asian Journal of Andrology ; (6): 345-350, 2019.
Article in English | WPRIM | ID: wpr-1009652

ABSTRACT

While it is known that spermatogonial stem cells (SSCs) initiate the production of male germ cells, the mechanisms of SSC self-renewal, proliferation, and differentiation remain poorly understood. We have previously identified Strawberry Notch 1 (SBNO1), a vertebrate strawberry notch family protein, in the proteome profile for mouse SSC maturation and differentiation, revealing SBNO1 is associated with neonatal testicular development. To explore further the location and function of SBNO1 in the testes, we performed Sbno1 gene knockdown in mice to study the effects of SBNO1 on neonatal testicular and SSC development. Our results revealed that SBNO1 is required for neonatal testicular and SSC development in mice. Particularly, in vitro Sbno1 gene knockdown with morpholino oligonucleotides caused a reduction of SSCs and inactivation of the noncanonical Wnt pathway, through Jun N-terminal kinases. Our study suggests SBNO1 maintains SSCs by promoting the noncanonical Wnt pathway.


Subject(s)
Animals , Male , Mice , Adult Germline Stem Cells/metabolism , Cell Proliferation/physiology , Gene Knockdown Techniques , Proteome , Repressor Proteins/metabolism , Testis/metabolism , Wnt Signaling Pathway/physiology
19.
Chinese Journal of Practical Gynecology and Obstetrics ; (12): 321-324, 2019.
Article in Chinese | WPRIM | ID: wpr-816185

ABSTRACT

OBJECTIVE: To investigate the effect of endometrial thickness(EMT)on the day of hCG administration on preg⁃nancy outcome in the fresh in vitro fertilization/intracytoplasmic sperm injection(IVF/ICSI)cycle.METHODS:A retrospec⁃tive analysis was conducted of 3601 IVF/ICSI-ET cycles between January 1,2015 and December 31,2015 in eight repro⁃ductive centers.The endometrial thickness was measured on hCG injection day and the distribution of endometrial thick⁃ness and pregnancy outcome was drawn.Patients were divided into two groups based on the EMT:group A(289 cycles,EMT<8 mm),group B(3312 cycles,EMT≥duration of pregnancy and birth weight of single gestation were compared.RESULTS:Group A had significantly lower clini⁃cal pregnancy rate(46.0% vs.55.9%,P=0.001),live birth rates(35.3% vs. 47.0%,P=0.000)and higher pregnancy loss rate(23.3% vs.15.8%,P=0.024)compared with group B,while there was no significant difference in duration of pregnan⁃cy or birth weight of single gestation.Logistic regression analyses showed that clinical pregnancy rate(aOR=1.492,P=0.001)and live birth rate(aOR=1.621,P=0.000)increased in group B compared with group A,when correcting for the women age,BMI and transfer embryo numbers.CONCLUSION:The endometrial thickness on the day of hCG administra⁃tion affects the pregnancy outcome in the fresh IVF/ICSI cycle.When EMT<8 mm,the clinical pregnancy rate and live birth rate of IVF-ET are lower,and the patient should be well informed before making the decision of em⁃bryo transfer.However,endometrial thickness on the day of hCG administration does not affect the duration of preg⁃nancy and the birth weight of the fetus.

20.
Chinese Journal of Laboratory Medicine ; (12): 262-269, 2019.
Article in Chinese | WPRIM | ID: wpr-746279

ABSTRACT

Objective To explore the molecular pathogenesis of 3 Glanzmann's thrombasthenia pedigree by using bioinformatics software and provide evidence for in vitro experiments. Methods The genetic analysis of 3 pedigree diagnosed as Glanzmann's thrombasthenia was carried out. Clustalx-2.1 win software was used to analyze the conservatism of mutant sites in homologous sequences. Bioinformatics software such as PolyPhen-2, PROVEAN, SIFT and Mutationtaster was used to analyze the biological effect of mutation. SPDBV software constructed the molecular structure model of mutant protein and evaluated the influence of mutation on protein structure. Results The "new mutations" found in 3 Glanzmann's thrombasthenia pedigree were ITGA2B:c. 814G>C (p. Val272Leu), ITGA2B:c. 432G>A (p. Trp144Ter) and ACTN1:c. 2458A>G (p. Ile820Val). All three mutations were highly conserved among homologous species. Mutationtaster software showed that 3 new mutations were likely pathogenic. PolyPhen-2 and PROVEAN software showed ITGA2B p.Val272Leu and ACTN1 p.Ile820Val were benign and SIFT software showed that ITGA2B p. Val272Leu were likely pathogenic, while ACTN1 p. Ile820Val is benign. The result of SPDBV software showed that the Val272 of ITGA2B was transformed to Leu, neutralizing all the original hydrogen bond. The Trp144 of ITGA2B is transformed to Ter, resulting in the truncated proteins with only 113 amino acid residues. All these mutations affected the molecular structure of GPⅡb, resulting in a decrease ofGPⅡb/Ⅲa expression. When the Ile820 of ACTN1 is transformed to Val, onlyretained the hydrogen bond of Ile820 and Asp822, neutralized the rest hydrogen bond, whichaffected the molecular structure and protein function of ACTN1. Conclusion The mutations of ITGA2B:c.814G>C (p.VAL272LEU), ITGA2B:c.432G>A (p.Trp144Ter) and ACTN1:c.2458A>G (p.Ile820Val) are pathogenic.

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