Cost-effectiveness of mepolizumab for severe eosinophilic asthma in China.

OBJECTIVE: To evaluate the economic value of mepolizumab as an add-on therapy to the standard of care (SoC) for patients with severe eosinophilic asthma in China. METHODS: A Markov model with three health conditions was constructed to calculate the incremental cost per quality-adjusted life year (QALY) in mepolizumab with SoC and SoC only groups from the perspective of the Chinese healthcare system throughout an entire lifespan. The model was populated with local costs, while efficacy parameters were obtained from the global Phase III MENSA trial and mortality was derived from two surveys. One-way and probabilistic sensitivity analyses were conducted. Additional scenario analysis was used to estimate the cost-effectiveness impact of changes in the price of mepolizumab. RESULTS: Over the lifetime treatment horizon, the incremental cost-effectiveness ratio (ICER) of mepolizumab plus SoC compared to SoC alone was $170 648.73 per QALY. Sensitivity analyses focused on these results. Scenario analysis showed that mepolizumab would require a price reduction of at least 82% to reach the current willingness-to-pay (WTP=$38 223.34/QALY) threshold. CONCLUSION: Mepolizumab is not a cost-effective healthcare resource in China at its current pricing.

New Advances in the Use of 18F-FET PET in the Diagnosis and Management of Adult High-Grade Gliomas / 罕见病研究

Gliomas are the most common primary intracranial tumors in adults, among which high-grade glioma patients are characterized by short survival and poor prognosis. The diagnosis, treatment, evaluation of effective treatments, and prognosis prediction of high-grade gliomas are of great significance for improving patient survival. Conventional enhanced magnetic resonance imaging has deficiencies in delineating tumor extent, identifying tumor progression and treatment-related changes. Therefore, there is a broad consensus to incorporate amino acid PET, and 18F-FET PET inparticular, into the diagnostic and therapeutic process of high-grade gliomas. In this article, we review the new research progress of 18F-FET PET in the diagnosis and treatment of adult high-grade glioma in recent years.

New Advances in the Use of 18F-FET PET in the Diagnosis and Management of Adult High-Grade Gliomas / 罕见病研究

Gliomas are the most common primary intracranial tumors in adults, among which high-grade glioma patients are characterized by short survival and poor prognosis. The diagnosis, treatment, evaluation of effective treatments, and prognosis prediction of high-grade gliomas are of great significance for improving patient survival. Conventional enhanced magnetic resonance imaging has deficiencies in delineating tumor extent, identifying tumor progression and treatment-related changes. Therefore, there is a broad consensus to incorporate amino acid PET, and 18F-FET PET inparticular, into the diagnostic and therapeutic process of high-grade gliomas. In this article, we review the new research progress of 18F-FET PET in the diagnosis and treatment of adult high-grade glioma in recent years.

CRISPR-assisted transcription activation by phase-separation proteins

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has been widely used for genome engineering and transcriptional regulation in many different organisms. Current CRISPR-activation (CRISPRa) platforms often require multiple components because of inefficient transcriptional activation. Here, we fused different phase-separation proteins to dCas9-VPR (dCas9-VP64-P65-RTA) and observed robust increases in transcriptional activation efficiency. Notably, human NUP98 (nucleoporin 98) and FUS (fused in sarcoma) IDR domains were best at enhancing dCas9-VPR activity, with dCas9-VPR-FUS IDR (VPRF) outperforming the other CRISPRa systems tested in this study in both activation efficiency and system simplicity. dCas9-VPRF overcomes the target strand bias and widens gRNA designing windows without affecting the off-target effect of dCas9-VPR. These findings demonstrate the feasibility of using phase-separation proteins to assist in the regulation of gene expression and support the broad appeal of the dCas9-VPRF system in basic and clinical applications.

Effects of Moxibustion on NLRP3 Inflammasome and Caspase-12 in Synovium of Experimental RA Rabbits / 世界科学技术-中医药现代化

Objective To observe the effects of moxibustion on the expression of Nucleotide binding oligomeric domain like receptor thermoprotein domain associated protein 3(Nod-like receptor protein 3,NLRP3)inflammasome and Cysteine aspartate protease 12(Caspase-12)in synovial tissue of experimental Rheumatoid Arthritis(RA)rabbits,and explore the anti-inflammatory mechanism of moxibustion on RA rabbits.Methods Twenty-four Japanese big-eared white rabbits were randomly divided into 3 groups according to body weight and sex:the control group,the model group and the moxibustion group.The Freund's Complete Adjuvant(FCA)was injected into the joint cavities of two knee joints of rabbits at the dose of 0.5 mL·kg-1 in the model group and the moxibustion group,and the control group was injected with the same amount of normal saline as control.For the moxibustion group,scar free moxibustion was operated at bilateral"BL 23"and"ST 36"acupoints,moxibustion for 3 times per acupoint per day,continuous treatment for 6 days,rest for 1 day,as a course of treatment,a total of 3 courses of treatment.The histological changes of synovium samples of rabbit knee joints were observed under light microscope,real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of NLRP3,Cysteine aspartate protease 1(Caspase-1),Caspase-12 in synovial tissues and Enzyme-linked immuno sorbent assay(ELISA)was used to measure the change of Interleukin-1β(IL-1β)content in the synovial fluid of experimental animals.Results Compared with the control group,the degree of synovial lesions(inflammatory cell infiltration,synovial tissue hyperplasia,fibrous tissue hyperplasia,macrophage hyperplasia,etc.)and pathological score were increased(P<0.05),the expression of NLRP3,Caspase-1 mRNA were increased(P<0.05),the expression of Caspase-12 mRNA was significantly downregulated(P<0.01)and the expression content of IL-1β was significantly up-regulated(P<0.01)in the model group;Compared with the model group,the synovial lesion degree and pathological score were decreased(P<0.05),the expression of NLRP3,Caspase-1 mRNA were decreased(P<0.05),the expression of Caspase-12 mRNA was significantly increased(P<0.01)and the expression of IL-1β was decreased(P<0.05)in the moxibustion group.Conclusion Moxibustion can significantly inhibit the pathological process of knee synovitis in RA rabbits;Moxibustion at"Shenshu(BL 23)"and"Zusanli(ST 36)"acupoints can significantly up-regulate the expression of Caspase-12 and then inhibit the NLRP3 inflammasome,which may be one of mechanisms of moxibustion to reduce RA inflammatory response.

Study on the Effect of Lentivirus-Mediated NLRP3 Overexpression on Levels of Anti-CCP and RF in Synovial Fluid of Experimental RA Rabbits Regulated by Moxibustion / 世界科学技术-中医药现代化

Objective Observation of the anti-inflammatory and detumescence effects on experimental rheumatoid arthritis(RA)rabbits by moxibustion,With lentiviral vector mediated NLRP3 overexpression in RA animal models,to investigate the effect of moxibustion on anti-CCP/RF levels in synovial fluid of RA rabbit joints and its regulation mechanism of NLRP3.Methods Randomly divided thirty Japanese big-ear white rabbits into 5 groups:group of blank,group of model,group of moxibustion,NLRP3 overexpression group and NLRP3 overexpression negative group,with 6 rabbits in each group.Both joint of the model group,and the groups with moxibustion was injected into FCA(0.5 mL·kg-1),the blank group used the same method to inject sterile saline.On the 3rd day and the 10th day after treatment,10 μL of NLRP3 overexpression lentiviral vector was injected into the joint cavity of the NLRP3 overexpression group of rabbits.The rabbits in the NLRP3 overexpression negative group were injected with the same amount of LvGFP as Contrast.moxibustion bilateral"BL23","ST 36"of the groups with moxibustion(5 cones with one point in a day.6 days for 1 course,1 day for rest,3 courses of treatment in total.)After treatment,observe the circumference of each rabbit's left and right knee joints,the content of RF,anti-CCP,IL-18 in synovial fluid was detected by ELISA.Results Compared with the blank group,the left and right knee joint circumferences in the model group increased,and the anti-CCP,RF,and IL-18 contents in the synovial fluid of the knee joint increased significantly(P<0.01);compared with the modle group,The circumference of the knee joint decreased,and the content of anti-CCP,RF and IL-18 decreased significantly(P<0.01);compared with the moxibustion group,the circumference of the left and right knee joints in the NLRP3 overexpression group increased(P<0.05),the content of anti-CCP,RF and IL-18 increased(P<0.05).Conclusions Moxibustion has anti-inflammatory effects on experimental RA rabbits.Moxibustion at Shenshu(BL23)and Zusanli(ST36)can significantly reduce the level of anti-CCP,RF the immune effector,also IL-18,the in synovial fluid of RA rabbits knee joint,yet this effect can be declined significantly under NLRP3 overexpression,suggesting that moxibustion performed its anti-inflammatory regulation by suppressing abnormal immune function in experimental RA rabbits,which may be closely related to the NLRP3 expression.

Weakly supervised multitask learning models to identify symptom onset time of unclear-onset intracerebral hemorrhage.

BACKGROUND: Approximately one-third of spontaneous intracerebral hemorrhage patients did not know the onset time and were excluded from studies about time-dependent treatments for hyperacute spontaneous intracerebral hemorrhage. AIMS: To help clinicians explore the benefit of time-dependent treatments for unclear-onset patients, we presented artificial intelligence models to identify onset time using non-contrast computed tomography (NCCT) based on weakly supervised multitask learning (WS-MTL) structure. METHODS: The patients with reliable symptom onset time (strong label) or repeat CT (weak label) were included and split into training set and test set (internal and external). The WS-MTL structure utilized strong and weak labels simultaneously to improve performance. The models included three binary classification models for classifying whether NCCT acquired within 6, 8 or 12 h for different treatments measured by area under curve, and a regression model for determining the exact onset time measured by mean absolute error. The generalizability of models was also explored in comprehensive analysis. RESULTS: A total of 4004 patients with 10,780 NCCT scans were included. The performance of WS-MTL classification model showed high accuracy, and that of regression model was satisfactory in ≤6 h subgroup. In comprehensive analysis, the WS-MTL showed better performance for larger hematomas and thinner scans. And the performance improved effectively as training amounts increasing and could be improved steadily through retraining. CONCLUSIONS: The WS-MTL models showed good performance and generalizability. Considering the large number of unclear-onset spontaneous intracerebral hemorrhage patients, it may be worth to integrate the WS-MTL model into clinical practice to identify the onset time.

Systemic lupus erythematosus combined with Aspergillus fumigatus brain abscess in 2 cases / 中华神经科杂志

Systemic lupus erythematosus (SLE) complicated with Aspergillus fumigatus brain abscess is rare and needs to be differentiated from bacterial brain abscess and neuropsychiatric lupus. This article reports 2 cases of surgically diagnosed SLE combined with Aspergillus fumigatus brain abscess. The first patient was a 34-year-old woman. Six months after the diagnosis of SLE, she developed convulsions and unconsciousness. She was diagnosed as neuropsychiatric lupus at the first hospitalization because of negative imaging. After discharge, repeated head magnetic resonance imaging revealed abscess-like signals. The second patient, a 20-year-old male, developed high fever, convulsions, and unconsciousness 3 years after the diagnosis of SLE, and head imaging showed an abscess-like signal. The etiology of the cerebrospinal fluid of the 2 patients was both negative, and the Aspergillus fumigatus brain abscess was diagnosed by pathology through abscess resection or drainage. After treatment with voriconazole, the symptoms were relieved and the lesions were subsided.

The Patterns of Morphological Change During Intracerebral Hemorrhage Expansion: A Multicenter Retrospective Cohort Study.

Objectives: Hemorrhage expansion (HE) is a common and serious condition in patients with intracerebral hemorrhage (ICH). In contrast to the volume changes, little is known about the morphological changes that occur during HE. We developed a novel method to explore the patterns of morphological change and investigate the clinical significance of this change in ICH patients. Methods: The morphological changes in the hematomas of ICH patients with available paired non-contrast CT data were described in quantitative terms, including the diameters of each hematoma in three dimensions, the longitudinal axis type, the surface regularity (SR) index, the length and direction changes of the diameters, and the distance and direction of movement of the center of the hematoma. The patterns were explored by descriptive analysis and difference analysis in subgroups. We also established a prognostic nomogram model for poor outcomes in ICH patients using both morphological changes and clinical parameters. Results: A total of 1,094 eligible patients from four medical centers met the inclusion criteria. In 266 (24.3%) cases, the hematomas enlarged; the median absolute increase in volume was 14.0 [interquartile range (IQR), 17.9] mL. The initial hematomas tended to have a more irregular shape, reflected by a larger surface regularity index, than the developed hematomas. In subtentorial and deep supratentorial hematomas, the center moved in the direction of gravity. The distance of center movement and the length changes of the diameters were small, with median values of less than 4 mm. The most common longitudinal axis type was anterior-posterior (64.7%), and the axis type did not change between initial and repeat imaging in most patients (95.2%). A prognostic nomogram model including lateral expansion, a parameter of morphological change, showed good performance in predicting poor clinical outcomes in ICH patients. Conclusions: The present study provides a morphological perspective on HE using a novel automatic approach. We identified certain patterns of morphological change in HE, and we believe that some morphological change parameters could help physicians predict the prognosis of ICH patients.

Efficacies of bevacizumab-related therapies in recurrent high-grade glioma: a single-center study / 中华神经医学杂志

Objective:To explore the efficacies of bevacizumab monotherapy and combination therapy of bevacizumab with irinotecan, semustine and cisplatin in patients with recurrent high-grade glioma.Methods:Seventy patients with recurrent high-grade glioma admitted to our hospital from January 2011 to November 2019 were chosen in our study; 38 patients received bevacizumab monotherapy, 13 patients accepted bevacizumab and semustine combination therapy, 11 patients received bevacizumab and cisplatin combination therapy, and 8 patients accepted bevacizumab and irinotecan combination therapy. Survival statuses (progression-free survival [PFS] and overall survival [OS]) of these patients were retrospectively analyzed.Results:The median OS and median PFS of the enrolled patients were 12.83 months and 6.23 months, respectively. The median OS and median PFS of patients accepted bevacizumab monotherapy were 10.92 months and 5.03 months, respectively. The median OS and median PFS of patients accepted bevacizumab and semustine combination therapy were 16.30 months and 6.77 months, respectively. The median OS in patients accepted bevacizumab and irinotecan combination therapy and patients accepted bevacizumab and cisplatin combination therapy was 11.90 months and 14.40 months, respectively.Conclusion:Bevacizumab by different therapy methods enjoys good efficacy; bevacizumab monotherapy or combination therapy can be recommended for recurrent high-grade glioma.

Live cell imaging and proteomic profiling of endogenous NEAT1 lncRNA by CRISPR/Cas9-mediated knock-in

In mammalian cells, long noncoding RNAs (lncRNAs) form complexes with proteins to execute various biological functions such as gene transcription, RNA processing and other signaling activities. However, methods to track endogenous lncRNA dynamics in live cells and screen for lncRNA interacting proteins are limited. Here, we report the development of CERTIS (CRISPR-mediated Endogenous lncRNA Tracking and Immunoprecipitation System) to visualize and isolate endogenous lncRNA, by precisely inserting a 24-repeat MS2 tag into the distal end of lncRNA locus through the CRISPR/Cas9 technology. In this study, we show that CERTIS effectively labeled the paraspeckle lncRNA NEAT1 without disturbing its physiological properties and could monitor the endogenous expression variation of NEAT1. In addition, CERTIS displayed superior performance on both short- and long-term tracking of NEAT1 dynamics in live cells. We found that NEAT1 and paraspeckles were sensitive to topoisomerase I specific inhibitors. Moreover, RNA Immunoprecipitation (RIP) of the MS2-tagged NEAT1 lncRNA successfully revealed several new protein components of paraspeckle. Our results support CERTIS as a tool suitable to track both spatial and temporal lncRNA regulation in live cells as well as study the lncRNA-protein interactomes.

Live cell imaging and proteomic profiling of endogenous NEAT1 lncRNA by CRISPR/Cas9-mediated knock-in

In mammalian cells, long noncoding RNAs (lncRNAs) form complexes with proteins to execute various biological functions such as gene transcription, RNA processing and other signaling activities. However, methods to track endogenous lncRNA dynamics in live cells and screen for lncRNA interacting proteins are limited. Here, we report the development of CERTIS (CRISPR-mediated Endogenous lncRNA Tracking and Immunoprecipitation System) to visualize and isolate endogenous lncRNA, by precisely inserting a 24-repeat MS2 tag into the distal end of lncRNA locus through the CRISPR/Cas9 technology. In this study, we show that CERTIS effectively labeled the paraspeckle lncRNA NEAT1 without disturbing its physiological properties and could monitor the endogenous expression variation of NEAT1. In addition, CERTIS displayed superior performance on both short- and long-term tracking of NEAT1 dynamics in live cells. We found that NEAT1 and paraspeckles were sensitive to topoisomerase I specific inhibitors. Moreover, RNA Immunoprecipitation (RIP) of the MS2-tagged NEAT1 lncRNA successfully revealed several new protein components of paraspeckle. Our results support CERTIS as a tool suitable to track both spatial and temporal lncRNA regulation in live cells as well as study the lncRNA-protein interactomes.

Live cell imaging and proteomic profiling of endogenous NEAT1 lncRNA by CRISPR/Cas9-mediated knock-in

In mammalian cells, long noncoding RNAs (lncRNAs) form complexes with proteins to execute various biological functions such as gene transcription, RNA processing and other signaling activities. However, methods to track endogenous lncRNA dynamics in live cells and screen for lncRNA interacting proteins are limited. Here, we report the development of CERTIS (CRISPR-mediated Endogenous lncRNA Tracking and Immunoprecipitation System) to visualize and isolate endogenous lncRNA, by precisely inserting a 24-repeat MS2 tag into the distal end of lncRNA locus through the CRISPR/Cas9 technology. In this study, we show that CERTIS effectively labeled the paraspeckle lncRNA NEAT1 without disturbing its physiological properties and could monitor the endogenous expression variation of NEAT1. In addition, CERTIS displayed superior performance on both short- and long-term tracking of NEAT1 dynamics in live cells. We found that NEAT1 and paraspeckles were sensitive to topoisomerase I specific inhibitors. Moreover, RNA Immunoprecipitation (RIP) of the MS2-tagged NEAT1 lncRNA successfully revealed several new protein components of paraspeckle. Our results support CERTIS as a tool suitable to track both spatial and temporal lncRNA regulation in live cells as well as study the lncRNA-protein interactomes.

Etiology and risk factors for meningitis and/or bacteremia after craniotomy for patients with glioma / 中南大学学报(医学版)

Objective:To evaluate the incidence and microbiological profile of meningitis and/or bacteremia after craniotomy in patients with glioma and analyze the risk factors relevant to postoperative meningitis.Methods:All demographic data,etiological data and clinical data for hospitalized patients who underwent craniotomy in Dept.of Neurosurgery of Peking Union Medical College Hospital between Jan.2013 to Dec.2017 were recorded.Results:The incidence of post-craniotomy meningitiswas 7.73％,bacteremia was 2.21％ and meningitis combined with bacteremia was 1.10％.The positive rate for cerebrospinal fluid culture was 78.57％.Among the 11 cases of meningitis,20 bacteria were detected,and the most common gram-positive bacteria were:epidermal Staphylococcus (3 case,15.0％),and hemolytic Staphylococcus (3 case,15.0％);the most common gram-negative one was Klebsiella pneumoniae (2 case,10.0％).Among the 5 cases of bacteremia,Klebsiella pneumoniae was the most common one (3 cases,60.0％).Most of the gram-positive bacteria were sensitive to vancomycin and were resistant to penicillin G.Most of the gram-negative bacteria were sensitive to the 3rd generation of cephalosporin and meropenem.Among all the 16 patients,13 were good in prognosis (81.3％) while 3 (18.8％) were poor in prognosis,with 2 deaths and 1 discharged automatically.Among the 11 patients with meningitis,7 patients (63.6％) were male and 7(63.6％) were older than 50 years old.The lesionsin the 6 patients located in the frontal lobe (54.5％) and the glioma in 6 patients was at WHO grade Ⅳ (54.5％).Three patients were diagnosed as diabetes before the surgery (27.3％).Conclusion:Postoperative meningitis and bacteremia can occur in patients with glioma.The pathogens and drug susceptibility are different.The risk factors for meningitis after craniotomy should be concerned before making a treatment plan.

Effects of moxibustion at different time points on foot swelling, serum level of TNF-α and circadian rhythm in rats with rheumatoid arthritis / 中国针灸

OBJECTIVE@#To study the effects of moxibustion at different time points on serum level of tumor necrosis factor-α (TNF-α) in rats with rheumatoid arthritis (RA), and to explore its regulation mechanism on circadian rhythm.@*METHODS@#A total of 96 Sprague-Dawley (SD) adult rats were randomly assigned into a blank group, a model group, a moxibustion at 5-7 AM group and a moxibustion at 5-7 PM group, 24 rats in each group, half male and half female. Each group was further divided into a 0 AM group, a 6 AM group, a 12 N group and a 6 PM group, 6 rats in each group. All rats were treated with the 12 h/12 h light-dark cycle in the whole process of experiment. Except for the blank group, all rats were treated with intracutaneous injection of freund's complete adjuvant (FCA) at right foot pad to establish the RA model. The rats at the two moxibustion groups were treated with grain-sized moxibustion at "Shenshu" (BL 23) and "Zusanli" (ST 36) at 5-7 AM and 5-7 PM, respectively, one side per treatment, once a day; six treatments were taken as one course and 3 courses were given with an interval of one day between courses. The rats in the remaining groups were treated with identical fixation but without moxibustion intervention. The right foot volume was measured before model establishment, after model establishment and after treatment. The blood samples were collected after treatment and the serum level of TNF-α was measured by ELISA. The SPSS 21.0 software and Halberg Cosinor were adopted to analyze the experiment data.@*RESULTS@#After treatment, compared with the blank group, the foot swelling severity was significantly increased in the model group, moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (all <0.01); compared with the model group, the foot swelling severity was significantly reduced in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (both <0.01). Compared with the blank group, the serum level of TNF-α was increased significantly in the model group and moxibustion at 5-7 AM group (both <0.05); compared with the model group, the serum level of TNF-α was reduced significantly in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (both <0.05). The serum level of TNF-α showed circadian rhythm in all the groups (all <0.05), and the peak appeared at night phase; compared with the blank group, the median value of TNF-α was increased significantly in the model group (<0.05), the peak phase was delayed and the amplitude was increased (<0.05); compared with the model group, the median value of TNF-α was significantly reduced in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (<0.01), the peak phase was advanced and the amplitude was reduced (<0.05).@*CONCLUSION@#Moxibustion could effectively reduce the serum level of TNF-α to relieve the foot swelling severity in RA rats. Moxibustion could regulate the circadian rhythm of TNF-α to play its effects on the inhibition of the synthesis of TNF-α. No efficacy is observed between the treatment at 5-7 AM and 5-7 PM.

DeepNitro: Prediction of Protein Nitration and Nitrosylation Sites by Deep Learning / 基因组蛋白质组与生物信息学报·英文版

Protein nitration and nitrosylation are essential post-translational modifications (PTMs) involved in many fundamental cellular processes. Recent studies have revealed that excessive levels of nitration and nitrosylation in some critical proteins are linked to numerous chronic diseases. Therefore, the identification of substrates that undergo such modifications in a site-specific manner is an important research topic in the community and will provide candidates for targeted therapy. In this study, we aimed to develop a computational tool for predicting nitration and nitrosylation sites in proteins. We first constructed four types of encoding features, including positional amino acid distributions, sequence contextual dependencies, physicochemical properties, and position-specific scoring features, to represent the modified residues. Based on these encoding features, we established a predictor called DeepNitro using deep learning methods for predicting protein nitration and nitrosylation. Using n-fold cross-validation, our evaluation shows great AUC values for DeepNitro, 0.65 for tyrosine nitration, 0.80 for tryptophan nitration, and 0.70 for cysteine nitrosylation, respectively, demonstrating the robustness and reliability of our tool. Also, when tested in the independent dataset, DeepNitro is substantially superior to other similar tools with a 7%-42% improvement in the prediction performance. Taken together, the application of deep learning method and novel encoding schemes, especially the position-specific scoring feature, greatly improves the accuracy of nitration and nitrosylation site prediction and may facilitate the prediction of other PTM sites. DeepNitro is implemented in JAVA and PHP and is freely available for academic research at http://deepnitro.renlab.org.

Effects and possible mechanisms of nerve growth factor on serum levels of VEGF and SDF-1 protein expression in rats with focal cerebral ischemia / 实用医学杂志

Objective To investigate the effects and the possible mechanisms of nerve growth factor (NGF) on serum levels of vascular endothelial growth factor (VEGF) and stromal cell-derived factor 1 (SDF-1) protein expression in rats with focal cerebral ischemia.Methods Male Sprague-Dawley rats weighting 200 ～ 250 g were subjected to middle cerebral occlusion (MCAO).MCAO rats were randomly divided into NGF group,saline control group and NGF+PI3K antagonist group (n =12).The neurological function was evaluated on the 4th,7th day after MCAO,and the serum levels of VEGF and SDF-1 protein were measured by ELISA.Results The neurological function was better in rats of the NGF group than those of the saline control group and NGF+PI3K antagonist on the 4th,7th day after MCAO (P ＜ 0.05).The serum levels of VEGF and SDF-1 protein of the NGF group was significantly higher than that of the saline control group and NGF+PI3K antagonist group on the 4th day after MCAO (P ＜ 0.05).Conclusions Treatment with NGF may improve neurological function of rats with focal cerebral ischemia,and upregulate serum levels of VEGF and SDF-1 protein expression.PI3K/AKT signal pathway may have attended the above regulation.

Correction of β-thalassemia mutant by base editor in human embryos

β-Thalassemia is a global health issue, caused by mutations in the HBB gene. Among these mutations, HBB -28 (A>G) mutations is one of the three most common mutations in China and Southeast Asia patients with β-thalassemia. Correcting this mutation in human embryos may prevent the disease being passed onto future generations and cure anemia. Here we report the first study using base editor (BE) system to correct disease mutant in human embryos. Firstly, we produced a 293T cell line with an exogenous HBB -28 (A>G) mutant fragment for gRNAs and targeting efficiency evaluation. Then we collected primary skin fibroblast cells from a β-thalassemia patient with HBB -28 (A>G) homozygous mutation. Data showed that base editor could precisely correct HBB -28 (A>G) mutation in the patient's primary cells. To model homozygous mutation disease embryos, we constructed nuclear transfer embryos by fusing the lymphocyte or skin fibroblast cells with enucleated in vitro matured (IVM) oocytes. Notably, the gene correction efficiency was over 23.0% in these embryos by base editor. Although these embryos were still mosaic, the percentage of repaired blastomeres was over 20.0%. In addition, we found that base editor variants, with narrowed deamination window, could promote G-to-A conversion at HBB -28 site precisely in human embryos. Collectively, this study demonstrated the feasibility of curing genetic disease in human somatic cells and embryos by base editor system.

Effective gene editing by high-fidelity base editor 2 in mouse zygotes

Targeted point mutagenesis through homologous recombination has been widely used in genetic studies and holds considerable promise for repairing disease-causing mutations in patients. However, problems such as mosaicism and low mutagenesis efficiency continue to pose challenges to clinical application of such approaches. Recently, a base editor (BE) system built on cytidine (C) deaminase and CRISPR/Cas9 technology was developed as an alternative method for targeted point mutagenesis in plant, yeast, and human cells. Base editors convert C in the deamination window to thymidine (T) efficiently, however, it remains unclear whether targeted base editing in mouse embryos is feasible. In this report, we generated a modified high-fidelity version of base editor 2 (HF2-BE2), and investigated its base editing efficacy in mouse embryos. We found that HF2-BE2 could convert C to T efficiently, with up to 100% biallelic mutation efficiency in mouse embryos. Unlike BE3, HF2-BE2 could convert C to T on both the target and non-target strand, expanding the editing scope of base editors. Surprisingly, we found HF2-BE2 could also deaminate C that was proximal to the gRNA-binding region. Taken together, our work demonstrates the feasibility of generating point mutations in mouse by base editing, and underscores the need to carefully optimize base editing systems in order to eliminate proximal-site deamination.

Effects of grain-sized moxibustion from 7 am to 9 am on circadian rhythm of inflammatory factor IL-6 in rats with rheumatoid arthritis / 中国针灸

<p><b>OBJECTIVE</b>To explore the rhythm regulatory mechanism of interleukin-6 (IL-6) in the process of moxibustion for rheumatoid arthritis (RA).</p><p><b>METHODS</b>A total of 144 Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group, a moxibustion group, a sham operation group, an operation group, an operation+moxibustion group, 24 rats in each one. Each group was divided into 4 time points (0:00 am, 6:00' am, 12:00 am, 6:00 pm), 6 rats in each time point. The Light-Dark 12 : 12 was given in all rats for light-dark cycle. Except the blank group, rats in the remaining groups were treated with intracutaneous injection of freund's complete adjuvant at right-side foot to establish the model of RA. After the model establishment, bilateral adrenal, glands were removed in the operation group and operation + moxibustion group, while those in the sham operation group were not removed with identical operation procedure. Rats in the moxibustion group and operation + moxibustion group were treated with grain-sized moxibustion from 7:00 am to 9:00 am at "Shenshu" (BL 23) and "Zusanli" (ST 36) once everyday, 6 times were taken as one session and 3 sessions were required tatclly, while rats in the remaining groups received identical fixation without moxibustion. The general health state and foot volume of rats were measured before model establishment, after establishment and after treatment. After treatment, rats were sacrificed at each time point to collect the blood sample and measure the content of IL-6 by using enzymne-immunoassay method.</p><p><b>RESULTS</b>Compared with the blank group, the foot swelling in the model group was obviously increased (P<0. 05); the IL-6 maintained circadian rhythm (P<0. 05), but the peak phase had a backward trend, famplitude had an increased trend and the median was significantly lifted (P<0. 05). Compared with model group, !the foot swelling in the moxibustion group was obviously decreased (P<0. 05); the IL-6 maintained circadian. rhythm (P<0. 05), and the peak phase had a forward trend, amplitude had a decreased trend and the median was significantly reduced (P<0. 05). Compared with the moxibustion group, the foot swelling in the operation--moxibustion group was obviously increased (P < 0.05); the IL-6 maintained circadian rhythm (P < 0.5), but the peak phase moved forwrd, and the median was significantly elevated (P < 0.05).</p><p><b>CONCLUSION</b>The IL-6 in plasma maintains significant pathological circadian rhythm in RA rats; with the complete hypothalamic-pituitary-adrenal axis, moxibustion is likely to regulate the circadian rhythm of IL-6 to play an important role of anti-inflammatory effect in RA rats.</p>