Astrocytes regulate glial scar formation in cerebral ischemic stroke / 中国组织工程研究

BACKGROUND:Cerebral ischemic stroke is one of the main fatal and disabling diseases in the clinic,but only a few patients benefit from vascular recanalization in time,so it is urgent to explore new and effective therapy.As one of the critical pathological changes of ischemic stroke,the glial scar formed mainly by astrocytes is one major cause that hinders axonal regeneration and neurological recovery at the late stage of stroke. OBJECTIVE:To elucidate the pathological process and crucial signal regulatory mechanism of astrocytes in the formation of glial scar after ischemic stroke,as well as the potential therapeutic targets,to provide a theoretical reference for intervening astrocytic scar formation against ischemic stroke effectively,and novel strategies for promoting post-stroke rehabilitation. METHODS:The relevant articles published in CNKI,PubMed and Web of Science databases from 2010 to 2022 were retrieved.The search terms were"Ischemic stroke,Brain ischemi*,Cerebral ischemi*,Astrocyt*,Astroglia*,Glial scar,Gliosis,Astrogliosis"in Chinese and English.Finally,78 articles were included after screening and summarized. RESULTS AND CONCLUSION:(1)Astrocytes play an important role in the maintenance of central nervous system homeostasis.After ischemic stroke,astrocytes change from a resting state to an active state.According to the different severities of cerebral ischemic injury,astrocyte activation changes dynamically from swelling and proliferation to glial scar formation.(2)Mature astrocytes are stimulated to restart the cell cycle,then proliferate and migrate to lesions,which is the main source of the glial scar.Neural stem cells in the subventricular zone,neuron-glial antigen 2 precursor cells and ependymal precursor cells in the brain parenchyma can also differentiate into astrocytes.Endothelin-1,aquaporin 4,ciliary neurotrophic factor and connexins are involved in this process.In addition,chondroitin sulfate proteoglycan,as the main component of the extracellular matrix,forms the dense glial scar barrier with proliferated astrocytes,which hinders the polarization and extension of axons.(3)Activation or inhibition of crucial signal molecules involved in astrocyte activation,proliferation,migration and pro-inflammation functions regulate the glial scar formation.Transforming growth factor beta 1/Smad and Janus kinase/signal transducer and activator of transcription 3 are classical pathways related to astrogliosis,while receptor-interacting protein 1 kinase and glycogen synthase kinase 3β are significant molecules regulating the inflammatory response.However,there are relatively few studies on Smad ubiquitination regulatory factor 2 and Interleukin-17 and their downstream signaling pathways in glial scar formation,which are worthy of further exploration.(4)Drugs targeting astrogliosis-related signaling pathways,cell proliferation regulatory proteins and inflammatory factors effectively inhibit the formation of glial scar after cerebral ischemic stroke.Among them,the role of commonly used clinical drugs such as melatonin and valproic acid in regulating glial scar formation has been verified,which makes it possible to use drugs that inhibit glial scar formation to promote the recovery of neurological function in patients with stroke.(5)Considering the protective effects of glial scar in the acute phase,how to choose the appropriate intervention chance of drugs to maintain the protective effect of the glial scar while promoting nerve regeneration and repair in the local microenvironment is the direction of future efforts.

Expression and clinical value of miR-124 and miR-1976 in serum of patients with Parkinson's disease / 中华老年医学杂志

Objective:To investigate the expression and clinical significance of microRNA-124(miR-124)and microRNA-1976(miR-1976)in the serum of patients with Parkinson's disease(PD).Methods:A total of 58 patients with PD were selected from September 2020 to June 2022 and categorized as the PD group.The Unified Parkinson's Disease Rating Scale(UPDRS)score was used to divide the PD patients into two groups: those with a UPDRS score≤60(25 patients)and those with a UPDRS score >60(33 patients). The Hoehn-Yahr grading scale was used to grade the PD patients.Additionally, 30 healthy individuals who had undergone a physical examination during the same period were selected as the control group.After collecting the subjects' serum, we performed real-time fluorescent quantitative PCR(qRT-PCR)to detect the expressions of miR-124 and miR-1976 in the serum.Logistic regression analysis was employed to analyze the influencing factors, and the diagnostic significance of serum miR-124 and miR-1976 in PD patients was evaluated using the receiver operating characteristic(ROC)curve.To predict the target genes of miR-1976, we utilized several software including TargetScan and Mirtarbase.Results:Compared to the control group, the PD group showed a significant down-regulation of serum miR-124 expression[(1.49±0.36) vs.(1.02±0.32)]( t=8.85, P<0.001), while miR-1976 expression was sharply up-regulated[(0.98±0.30) vs.(1.33±0.37)]( t=6.92, P<0.001). The low expression of serum miR-124 and the overexpression of miR-1976 were identified as independent risk factors for PD( OR>1, P<0.05). The Hoehn-Yahr rating of PD patients with a UPDRS score above 60 was higher than that of patients with a UPDRS score below 60[(3.42 ± 0.73) vs.(2.16 ± 0.42)]( t=3.05, P<0.05). However, there was no significant difference in serum miR-124 and miR-1976 expression between groups with different UPDRS scores[miR-124: (1.09±0.26) vs.(0.98±0.38)( t=0.89, P>0.05); miR-1976: (1.42±0.43) vs.(1.23±0.68)( t=0.62, P>0.05)]. The ROC analysis results demonstrated that miR-124 and miR-1976 had area under the curve(AUC)values of 0.832 and 0.797, respectively, in diagnosing PD.The corresponding cutoff values were 1.205 and 1.196, respectively.The sensitivity for miR-124 was 74.1%, while for miR-1976 it was 51.8%.The specificity for miR-124 was 77.8%, and for miR-1976 it was 90.1%.When both miR-124 and miR-1976 were combined in the diagnosis of PD, the AUC was 0.912, with a sensitivity of 76.4% and a specificity of 93.2%.Furthermore, it was found that miR-1976 targeted the PINK1 gene, suggesting its potential as a target gene in PD. Conclusions:The expression of miR-124 was found to be decreased in PD patients, while the expression of miR-1976 was increased.Both miR-124 and miR-1976 showed some reference value in PD diagnosis, and their combined diagnostic value was higher.This suggests that further study on their significance is warranted.However, it should be noted that the expressions of miR-124 and miR-1976 were not found to be correlated with the UPDRS score of PD patients.

Smooth Pursuit and Reflexive Saccade in Discriminating Multiple-System Atrophy With Predominant Parkinsonism From Parkinson’s Disease

Background@#and Purpose Performing the differential diagnosis of Parkinson’s disease (PD) and multiple-system atrophy of parkinsonian type (MSA-P) is challenging. The oculomotor performances of patients with PD and MSA-P were investigated to explore their potential role as a biomarker for this differentiation. @*Methods@#Reflexive saccades and smooth pursuit were examined in 56 patients with PD and 34 with MSA-P in the off-medication state. @*Results@#Patients with PD and MSA-P had similar oculomotor abnormalities of prolonged and hypometric reflexive saccades. The incidence rates of decreased reflexive saccadic velocity and saccadic smooth pursuit were significantly higher in MSA-P than in PD (p<0.05 for both). Multiple logistic regression analysis indicated that slowed reflexive saccades (odds ratio [OR]=8.14, 95% confidence interval [CI]=1.45–45.5) and saccadic smooth pursuit (OR=5.27, 95% CI=1.24–22.43) were significantly related to MSA-P. @*Conclusions@#The distinctive oculomotor abnormalities of saccadic smooth pursuit and slowed reflexive saccades in MSA-P may serve as useful biomarkers for discriminating MSA-P from PD.

Establishment of a nomogram model for predicting liver cirrhosis with esophagogastric variceal bleeding based on aspartate aminotransferase-to-platelet ratio index and platelet-albumin-bilirubin score / 临床肝胆病杂志

ObjectiveTo investigate the value of aspartate aminotransferase-to-platelet ratio index （APRI） and platelet-albumin-bilirubin （PALBI） score in predicting the risk of esophagogastric variceal bleeding in patients with liver cirrhosis. MethodsA total of 119 patients with liver cirrhosis who were admitted to The First Affiliated Hospital of Soochow University from May 2021 and June 2022 were enrolled， and clinical data， routine blood test results， serum biochemistry， and coagulation test results were collected from all patients. According to the presence or absence of esophagogastric variceal bleeding， the patients were divided into non-bleeding group with 59 patients and bleeding group with 60 patients， and a comparative analysis was performed for the two groups. The independent samples t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； the chi-squared test or the Fisher’s exact test was used for comparison of categorical data between groups. The multivariate Logistic regression analysis was used to identify the independent risk factors for esophagogastric variceal bleeding in patients with liver cirrhosis and establish a nomogram predictive model. ResultsThe male patients accounted for 75.00% in the bleeding group and 40.68% in the non-bleeding group， and there was a significant difference in sex composition between the two groups （χ2=14.384， P<0.001）. Chronic hepatitis B was the main etiology in both the bleeding group and the non-bleeding group （53.33% vs 38.98%）， and there was no significant difference in composition ratio between the two groups （χ2=2.464， P=0.116）. Compared with the non-bleeding group， the bleeding group had a significantly higher activity of AT-IIIA （t=3.329， P=0.001） and significantly lower levels of PLT， TBil， Ca， TC， and TT （all P<0.05）. There were significant differences in APRI and PALBI between the two groups （χ2=6.175 and 19.532， both P<0.05）. The binary logistic regression analysis showed that APRI （odds ratio ［OR］=0.309， 95% confidence interval ［CI］： 0.109‍ ‍—‍ ‍0.881， P=0.028）， PALBI （OR=7.667， 95%CI： 2.005‍ ‍—‍ ‍29.327， P=0.003）， Ca （OR=0.001， 95%CI： 0.000‍ ‍—‍ ‍0.141， P=0.007）， TC （OR=0.469， 95%CI： 0.226‍ ‍—‍ ‍0.973， P=0.042）， and TT （OR=0.599， 95%CI： 0.433‍ ‍—‍ ‍0.830， P=0.002） were independent influencing factors for esophagogastric variceal bleeding in liver cirrhosis. A nomogram model was established based on the above factors and had an index of concordance of 0.899 and a well-fitted calibration curve. ConclusionAPRI and PALBI have a good value in predicting esophagogastric variceal bleeding in patients with liver cirrhosis， and the nomogram model established based on this study can predict the incidence rate of esophagogastric variceal bleeding in patients with liver cirrhosis.

Effect of Chaihu Guizhitang on Triple-negative Breast Cancer Cells： Based on HIF-1α/VEGFA Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Chaihu Guizhitang on triple-negative breast cancer （TNBC） cells based on hypoxia-inducible factor-1α （HIF-1α）/vascular endothelial growth factor A （VEGFA） signaling pathway. MethodTNBC xenograft model was established and the cells were randomized into model group， capecitabine group （0.2 mg·kg-1）， Chaihu Guizhitang low-dose group， medium-dose group， and high-dose group （10.62， 21.23， 42.46 g·kg-1）， with 10 mice in each group. After 21 days of medication， the content of tumor necrosis factor-α （TNF-α） in serum was detected by enzyme-linked immunosorbent assay （ELISA）. The expression of HIF-1α mRNA was detected by real-time fluorogenic quantitative polymerase chain reaction （real-time PCR）. Immunohistochemistry （IHC） was employed to detect the expression of HIF-1α， TNF-α， and VEGFA in tumor tissues， and CD34 staining to examine the angiogenesis in tumor tissues. Microvessel density （MVD） was calculated， and the protein expression of HIF-1α， VEGFA， and epidermal growth factor receptor （EGFR） in tumor tissues was measured by Western blot. ResultCompared with the model group， the rest four groups showed low levels of TNF-α （P<0.01）， HIF-1α mRNA （P<0.01）， expression of HIF-1α， TNF-α， VEGFA， and CD34 in cells， and MVD （P<0.05， P<0.01）， and low protein levels of HIF-1α， VEGFA， and EGFR （P<0.01）. Compared with capecitabine group， medium-dose and high-dose Chaihu Guizhitang decreased the level of TNF-α （P<0.01）， HIF-1α mRNA （P<0.01）， expression of HIF-1α， TNF-α， and VEGFA in cells （P<0.01）， CD34 expression， MVD， and protein levels of HIF-1α， VEGFA， and EGFR （P<0.01）. ConclusionChaihu Guizhitang may inhibit the angiogenesis in TNBC cells by regulating the expression of HIF-1α/VEGFA signaling pathway， thus exerting anti-tumor effect.

Clinical application of transcatheter arterial methylene blue angiography in the localization of lower gastrointestinal arterial bleeding / 介入放射学杂志

Objective To evaluate the clinical application value of transcatheter arterial methylene blue angiography in the localization of lower gastrointestinal arterial bleeding.Methods Ten patients with lower gastrointestinal arterial bleeding received interventional celiac artery angiography.After the bleeding responsible arteries were identified,a microcatheter was super-selectively placed in the bleeding responsible artery.During surgical procedure,the methylene blue solution was injected through the microcatheter to display the bleeding segment of the intestinal tract,providing precise localization of the bleeding intestinal segment for surgical resection.Results Transcatheter arterial methylene blue angiography could clearly display the bleeding segment of the intestinal tract.The bleeding segments of the intestinal tract in the 10 patients were quickly and accurately removed.After surgery,the gastrointestinal bleeding stopped,and no surgery-related complications occurred.Conclusion Transcatheter arterial methylene blue angiography can accurately detect the arterial bleeding segment of the lower gastrointestinal tract,which provides precise localization for quickly removing the bleeding segment of intestinal tract,therefor,this technique is worthy of widespread clinical application.(J Intervent Radiol,2023,32:1230-1232)

elationship between preoperative fasting plasma glucose and postoperative pulmonary complications after lung resection in type 2 diabetic patients / 中国胸心血管外科临床杂志

@#Objective     To explore the relationship between preoperative fasting plasma glucose (FPG) and postoperative pulmonary complications (PPCs) in type 2 diabetic patients undergoing elective thoracoscopic lung resection, and provide a reference for prediction and prevention of PPCs in the clinic. Methods     A retrospective analysis was performed on the type 2 diabetic patients who underwent elective thoracoscopic lung resection for the first time in our hospital from January 2017 to March 2021. According to the level of FPG one day before the operation, the patients were divided into three groups: a hypoglycemia group (<6.1 mmol/L), a medium level blood glucose group (≥6.1 mmol/L and <8.0 mmol/L) and a high blood glucose group (≥8.0 mmol/L). Besides, the patients were divided into a PPCs group and a non-PPCs group according to whether PPCs occurred. The risk factors for PPCs were analyzed by logistic regression analysis, and the predictive value of preoperative FPG level on PPCs was estimated by the area under the receiver operating characteristic curve (AUC). Results     A total of 130 patients were included, including 75 (57.7%) males and 55 (42.3%) females with an average age of 63.5±9.0 years. Logistic regression analysis showed that compared to non-PPCs patients, the level of preoperative FPG (P=0.023) and smoking history ratio (P=0.036) were higher and the operation time was longer (P=0.004) in the PPCs patients. High FPG level on preoperative day 1 and longer operation time were associated with PPCs risk. Besides, the preoperative FPG of 6.79 mmol/L was the threshold value to predict the occurrence of PPCs [AUC=0.653, 95%CI (0.559, 0.747), P=0.003]. Conclusion     There is a certain correlation between preoperative FPG level and postoperative PPCs, which may be used as an index to predict the occurrence of PPCs.

Discovery of the mechanisms of acupuncture in the treatment of migraine based on functional magnetic resonance imaging and omics / 医学前沿

Migraine is one of the most prevalent and disabling neurological disease, but the current pharmacotherapies show limited efficacy and often accompanied by adverse effects. Acupuncture is a promising complementary therapy, but further clinical evidence is needed. The influence of acupuncture on migraine is not an immediate effect, and its mechanism remains unclear. This study aims to provide further clinical evidence for the anti-migraine effects of acupuncture and explore the mechanism involved. A randomized controlled trial was performed among 10 normal controls and 38 migraineurs. The migraineurs were divided into blank control, sham acupuncture, and acupuncture groups. Patients were subjected to two courses of treatment, and each treatment lasted for 5 days, with an interval of 1 day between the two courses. The effectiveness of treatment was evaluated using pain questionnaire. The functional magnetic resonance imaging (fMRI) data were analyzed for investigating brain changes induced by treatments. Blood plasma was collected for metabolomics and proteomics studies. Correlation and mediation analyses were performed to investigate the interaction between clinical, fMRI and omics changes. Results showed that acupuncture effectively relieved migraine symptoms in a way different from sham acupuncture in terms of curative effect, affected brain regions, and signaling pathways. The anti-migraine mechanism involves a complex network related to the regulation of the response to hypoxic stress, reversal of brain energy imbalance, and regulation of inflammation. The brain regions of migraineurs affected by acupuncture include the lingual gyrus, default mode network, and cerebellum. The effect of acupuncture on patients' metabolites/proteins may precede that of the brain.

Mechanism of miR⁃148b⁃3p regulating proliferation of keloid derived fibroblasts / 安徽医科大学学报

Objective @#To analyze the effect of miR⁃148b⁃3p on the proliferation of keloid derived fibroblasts. @*Methods @#The expression levels of miR⁃148b⁃3p and SPARC in human keloid derived fibroblasts (HKF) and normal human fibroblasts (NFS) were analyzed by real time PCR. The expression level of SPARC protein was detected by Western blot. The effects of miR⁃148b⁃3p and SPARC on HKF proliferation were analyzed by CCK⁃8 method the luciferase reporter plasmid was constructed. The targeted binding site of miR⁃148b⁃3p and the target gene was analyzed by luciferase reporter gene method.@*Results @#miR⁃148b⁃3p was low expressed in HKF and SPARC was high expressed in HKF. Transfection of miR⁃148b⁃3p in HKF cells could down regulate the expression of SPARC and inhibit cell proliferation. Online analysis software predicted that miR⁃148b⁃3p could target the 3 ′⁃ UTR binding SPARC ; The results of dual luciferase reporter gene further confirmed that miR⁃148b⁃3p could target the 3 ′⁃ UTR of SPARC. Transfection of SPARC eukaryotic expression plasmid into HKF transfected with miR⁃148b⁃3p could counteract the effect of miR⁃148b⁃3p and restore cell proliferation. @*Conclusion @# miR⁃148b⁃3p can inhibit the proliferation of HKF by targeting the 3 ′⁃ UTR of SPARC and inhibiting its expression.

A high-efficiency and versatile CRISPR/Cas9-mediated HDR-based biallelic editing system / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9), the third-generation genome editing tool, has been favored because of its high efficiency and clear system composition. In this technology, the introduced double-strand breaks (DSBs) are mainly repaired by non-homologous end joining (NHEJ) or homology-directed repair (HDR) pathways. The high-fidelity HDR pathway is used for genome modification, which can introduce artificially controllable insertions, deletions, or substitutions carried by the donor templates. Although high-level knock-out can be easily achieved by NHEJ, accurate HDR-mediated knock-in remains a technical challenge. In most circumstances, although both alleles are broken by endonucleases, only one can be repaired by HDR, and the other one is usually recombined by NHEJ. For gene function studies or disease model establishment, biallelic editing to generate homozygous cell lines and homozygotes is needed to ensure consistent phenotypes. Thus, there is an urgent need for an efficient biallelic editing system. Here, we developed three pairs of integrated selection systems, where each of the two selection cassettes contained one drug-screening gene and one fluorescent marker. Flanked by homologous arms containing the mutated sequences, the selection cassettes were integrated into the target site, mediated by CRISPR/Cas9-induced HDR. Positively targeted cell clones were massively enriched by fluorescent microscopy after screening for drug resistance. We tested this novel method on the amyloid precursor protein (APP) and presenilin 1 (PSEN1) loci and demonstrated up to 82.0% biallelic editing efficiency after optimization. Our results indicate that this strategy can provide a new efficient approach for biallelic editing and lay a foundation for establishment of an easier and more efficient disease model.

Targeting a cryptic allosteric site of SIRT6 with small-molecule inhibitors that inhibit the migration of pancreatic cancer cells

SIRT6 belongs to the conserved NAD+-dependent deacetylase superfamily and mediates multiple biological and pathological processes. Targeting SIRT6 by allosteric modulators represents a novel direction for therapeutics, which can overcome the selectivity problem caused by the structural similarity of orthosteric sites among deacetylases. Here, developing a reversed allosteric strategy AlloReverse, we identified a cryptic allosteric site, Pocket Z, which was only induced by the bi-directional allosteric signal triggered upon orthosteric binding of NAD+. Based on Pocket Z, we discovered an SIRT6 allosteric inhibitor named JYQ-42. JYQ-42 selectively targets SIRT6 among other histone deacetylases and effectively inhibits SIRT6 deacetylation, with an IC50 of 2.33 μmol/L. JYQ-42 significantly suppresses SIRT6-mediated cancer cell migration and pro-inflammatory cytokine production. JYQ-42, to our knowledge, is the most potent and selective allosteric SIRT6 inhibitor. This study provides a novel strategy for allosteric drug design and will help in the challenging development of therapeutic agents that can selectively bind SIRT6.

Role of Yes-associated protein 1 in acute lung injury in septic mice: the relationship with ferroptosis / 中华麻醉学杂志

Objective:To evaluate the role of Yes-associated protein 1 (YAP1) in acute lung injury (ALI) and the relationship with ferroptosis in septic mice.Methods:Twenty-four male wild-type mice and 24 YAP1 conditional knockout mice, aged 9-10 weeks, weighing 22-25 g, were divided into 2 groups ( n=12 each) using a random number table method: wild-type sham operation group (WT+ Sham group) and wild-type sepsis-induced ALI group (WT+ ALI group); YAP1 conditional knockout sham operation group (CKO+ Sham group) and YAP1 conditional knockout sepsis-induced ALI group (CKO+ ALI group). The sepsis-induced ALI model was developed by cecal ligation and perforation (CLP) in anesthetized animals.The bronchoalveolar lavage fluid (BALF) was collected at 24 h after CLP to determine the protein concentration (by bicinchoninic acid method) and concentrations of interleukin-1beta (IL-1β) and tumor necrosis factor-α (TNF-α) (by enzyme-linked immunosorbent assay). Mice were then sacrificed, and the lung tissues were obtained for examination of ultrastructure (using a transmission electron microscope) and for determination of wet/dry lung weight ratio (W/D ratio), contents of Fe 2+ , malondialdehyde (MDA) and glutathione (GSH) (by colorimetric assay), and expression of YAP1, glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and solute carrier family 7 member 11 (SLC7A11) (by Western blot). Results:Compared with WT+ Sham group, the concentrations of protein in BALF, IL-1β and TNF-α were significantly increased, W/D ratio and contents of Fe 2+ and MDA were increased, GSH contents were decreased, the expression of GPX4 and SLC7A11 was down-regulated, ACSL4 expression was up-regulated ( P<0.05), alveolar epithelial cells showed characteristic changes of ferroptosis with mitochondrial shrinkage and decreased mitochondrial cristae in WT+ ALI group.Compared with WT+ CLP and CKO+ Sham groups, the concentrations of protein in BALF, IL-1β and TNF-α were significantly increased, W/D ratio and contents of Fe 2+ and MDA were increased, GSH contents were decreased, the expression of GPX4 and SLC7A11 was down-regulated, ACSL4 expression was up-regulated ( P<0.05), and the mitochondria in alveolar epithelial cells in lung tissues shrank obviously, and the mitochondrial cristae were reduced or even disappeared in CKO+ CLP group ( P<0.05). Conclusions:YAP1 is involved in the endogenous protective mechanism against ALI, which is related to inhibition of ferroptosis in septic mice.

Relationship between Nrf2 and ferroptosis during lung ischemia-reperfusion in rats / 中华麻醉学杂志

Objective:To evaluate the relationship between nuclear factor E2-related factor 2 (Nrf2) and ferroptosis during lung ischemia-reperfusion (I/R) in rats.Methods:Fifty-four healthy male Sprague-Dawley rats, weighing 220-250 g, were divided into 3 groups ( n=18 each) using a random number table method: sham operation group (Sham group), lung I/R group (IR group), and lung I/R+ Nrf2 agonist sulforaphane group (IR+ SFP group). Lung I/R model was developed by clamping the left pulmonary hilum for 60 min followed by 120 min of reperfusion.In IR+ SFP group, SFP 500 μg/kg was intraperitoneally injected at 3 days before lung ischemia once a day for 3 consecutive days, and the model was developed at 2 h after the end of administration.The rats were sacrificed at the end of reperfusion, and the bronchoalveolar lavage fluid (BALF) was collected to determine the protein concentration (using bicinchoninic acid method), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) concentrations (by enzyme-linked immunosorbent assay). The animals were then sacrificed, and lung tissue specimens were harvested for microscopic examination of the pathological changes (with a transmission electron microscope) and for determination of wet/dry weight (W/D) ratio, contents of iron, malondialdehyde (MDA) and glutathione (GSH) (by chemical colorimetric) and expression of nuclear Nrf2, glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4) (by Western blot). Results:Compared with Sham group, the concentrations of protein, IL-6 and TNF-α in BALF, W/D ratio, and contents of Fe 2+ and MDA were significantly increased, GSH content was decreased, GPX4 expression was down-regulated, the expression of nuclear Nrf2and ACSL4 was up-regulated ( P<0.05), and the mitochondrial morphology of type Ⅱalveolar epithelial cells showed the characteristic changes of ferroptosis, including the presence of smaller mitochondria and reduced cristae in IR group.Compared with IR group, the concentrations of protein, IL-6 and TNF-α in BALF, W/D ratio, and contents of Fe 2+ and MDA were significantly decreased, GSH content was increased, the expression of nuclear Nrf2 and GPX4 expression was up-regulated, ACSL4 expression was down-regulated ( P<0.05), and the pathological changes of lung tissues were significantly attenuated in IR+ SFP group. Conclusions:Nrf2 can inhibit ferroptosis during lung I/R and is involved in the endogenous protective mechanism in rats.

Deep learning: a new multifaceted tool assisting biomedical research and clinical diagnosis / 国际生物医学工程杂志

Artificial intelligence (AI) is one of the hottest research topics. The development of AI technology not only brings convenience to people's lives, but also integrates with other frontier fields to aid in data processing and result prediction. Deep learning is one of the emerging technologies that demonstrate outstanding performances. In this paper, the wide application of deep learning technology in many fields of biomedicine was summarized, common methods and models were briefly introduced including artificial neural network, deep neural network, convolutional neural network and recurrent neural network. Besides, the application of deep learning in biomedical image analysis, omics data processing and protein spatial structure prediction was summarized, and its limitations and development prospects in the above applications were briefly discussed.

Construction and verification of an intelligent measurement model for diabetic foot ulcer / 中南大学学报(医学版)

OBJECTIVES@#The measurement of diabetic foot ulcers is important for the success in diabetic foot ulcer management. At present, it lacks the accurate and convenient measurement tools in clinical. In recent years, artificial intelligence technology has demonstrated the potential application value in the field of image segmentation and recognition. This study aims to construct an intelligent measurement model of diabetic foot ulcers based on the deep learning method, and to conduct preliminary verification.@*METHODS@#The data of 1 042 diabetic foot ulcers clinical samples were collected. The ulcers and color areas were manually labeled, of which 782 were used as the training data set and 260 as the test data set. The Mask RCNN ulcer tissue color semantic segmentation and RetinaNet scale digital scale target detection were used to build a model. The training data set was input into the model and iterated. The test data set was used to verify the intelligent measurement model.@*RESULTS@#This study established an intelligent measurement model of diabetic foot ulcers based on deep learning. The mean average precision@.5 intersection over union (mAP@.5IOU) of the color region segmentation in the training set and the test set were 87.9% and 63.9%, respectively; the mAP@.5IOU of the ruler scale digital detection in the training set and the test set were 96.5% and 83.4%, respectively. Compared with the manual measurement result of the test sample, the average error of the intelligent measurement result was about 3 mm.@*CONCLUSIONS@#The intelligent measurement model has good accuracy and robustness in measuring the diabetic foot ulcers. Future research can further optimize the model with larger-scale data samples.

Targeting RAS phosphorylation in cancer therapy: Mechanisms and modulators

RAS, a member of the small GTPase family, functions as a binary switch by shifting between inactive GDP-loaded and active GTP-loaded state. RAS gain-of-function mutations are one of the leading causes in human oncogenesis, accounting for ∼19% of the global cancer burden. As a well-recognized target in malignancy, RAS has been intensively studied in the past decades. Despite the sustained efforts, many failures occurred in the earlier exploration and resulted in an 'undruggable' feature of RAS proteins. Phosphorylation at several residues has been recently determined as regulators for wild-type and mutated RAS proteins. Therefore, the development of RAS inhibitors directly targeting the RAS mutants or towards upstream regulatory kinases supplies a novel direction for tackling the anti-RAS difficulties. A better understanding of RAS phosphorylation can contribute to future therapeutic strategies. In this review, we comprehensively summarized the current advances in RAS phosphorylation and provided mechanistic insights into the signaling transduction of associated pathways. Importantly, the preclinical and clinical success in developing anti-RAS drugs targeting the upstream kinases and potential directions of harnessing allostery to target RAS phosphorylation sites were also discussed.

Clinical analysis of bone mineral density in patients with newly diagnosed untreated systemic lupus erythematous / 中华风湿病学杂志

Objective:The study was aimed to investigate the bone mineral density (BMD) status of newly diagnosed systemic lupus erythematous (SLE) patients.Methods:Newly diagnosed SLE patients and healthy controls in Peking University Third Hospital from 2014 to 2018 were enrolled into this cross-sectional study. Medical records including systemic lupus erythematosus disease activity index (SLEDAI)-2000 and BMD of the lumbar vertebrae and hips measured by dual-energy X-ray absorptiometry were collected. Patients were divided into normal and low BMD groups. Parameters were compared by Student- t test, Mann-Whitney U test and χ 2 test. Results:Eighty-nine patients and 20 healthy controls were included. Approximately 52.8% of the SLE patients had low BMD. Compared to the healthy control group, the BMD of lumbar spine and hip was obviously lower than in the newly diagnosed SLE group[(0.97±0.14) g/cm 2, (1.08±0.10) g/cm 2, t=3.548, P<0.01; (0.88±0.15) g/cm 2, (1.00±0.08) g/cm 2, t=3.878, P<0.01]. The BMD of lumbar spine and hip in the low BMD group was lower than that in the normal BMD group [(0.88±0.10) g/cm 2, (0.80±0.11) g/cm 2; (1.07±0.11) g/cm 2, (0.97±0.13) g/cm 2, respectively]. Compared with the normal BMD group, the body mass index (BMI) was significantly lower [(19.2±2.0) kg/m 2, (23.2±3.6) kg/m 2 respectively, t=3.678, P<0.01], the disease duration was longer [(45.7±7.7) weeks, (16.0±19.5) weeks, respectively, t=-3.306, P<0.01). Patients with low BMD tended to have lower 25-hydroxy-vitamin-D(25-OH-VD 3) level and higher bone metabolism marker levels (Degradation products of collagen B) [(9±5) nmol/L vs (12±7) nmol/L, t=1.385, P>0.05; 0.62(0.21, 1.61) ng/ml vs 0.43(0.19, 0.88) ng/ml, Z=0.624, P>0.05], although their differences didn't reach the statistical significant difference. Conclusion:Newly diagnosed SLE patients' BMD is lower than the healthy people. About 52.8% newly diagnosed untreated SLE patients have low BMD. Lower BMI and longer disease duration are their clinical characteristics. High bone metabolism marker levels, low 25-hydroxy-vitamin-D level might have clinical significance, although the current findings do not find statistically significant difference. Large sample size is required for subsequent research analysis.

Role of Erbin in muramyl dipeptide-induced inflammatory responses in macrophages of mice / 中华麻醉学杂志

Objective:To evaluate the role of ErbB2-interacting protein (Erbin) in muramyl dipeptide (MDP)-induced inflammatory responses in the macrophages of mice.Methods:Erbin gene knockout RAW264.7 cell line (Erbin -/ -RAW264.7) was constructed by CRISPR/CAS9 gene-editing technology.RAW264.7 cells were cultured in vitro.Each type of cells was divided into 2 groups ( n=16 each)by a random number table method: RAW264.7 group, RAW264.7 plus MDP group, erbin -/ -RAW264.7 group, and erbin -/ -RAW264.7 plus MDP group.In each MDP group, cells were incubated with 10 μg/ml MDP for 6 h, then immunofluorescence was used to determine the expression of nuclear factor kappa B (NF-κB) p65, and the concentrations of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the culture medium were determined by enzyme-linked immunosorbent assay. Results:Compared with RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased( P<0.05), NF-κB p65 moved to the nucleus, and the red fluorescence area was increased in RAW264.7+ MDP group.Compared with RAW264.7+ MDP group and Erbin -/- RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased ( P<0.05), NF-κB p65 moved more markedly to the nucleus, and the red fluorescence area was increased in Erbin -/-RAW264.7+ MDP group. Conclusion:Erbin inhibits MDP-induced inflammatory responses in macrophages through inhibiting the activity of NF-κB p65 in mice.

Effect of oxycodone hydrochloride injection on focal cerebral ischemia-reperfusion injury in rats / 中华急诊医学杂志

Objective:To investigate the effect of oxycodone hydrochloride injection pretreatment on focal cerebral ischemia-reperfusion injury in rats.Methods:Seventy-two male SD rats weighing 200-250 g were randomly divided into 3 groups( n=24 each group): sham operation group (sham group), focal cerebral I/R group (I/R group), and oxycodone hydrochloride injection group (Oxy group). Focal cerebral I/R was induced by middle cerebral artery occlusion for 2 h followed by reperfusion. In the Oxy group, oxycodone hydrochloride 0.5 mg/kg was injected iv at 5 min before ischemia. While the same volume of saline (1 mL) was injected in the sham group and I/R group. The neurological deficit score (NDS) was assessed at 24 h of reperfusion, the rats were then sacrificed, and their brains were immediately removed for determination of brain water content and the infarct volume, and the histopathological changes were observed after HE staining. The levels of cytokines (TNF-α, IL-1β and IL-10) in the ischemia cortex were quantified by ELISA. MPO activity in the ischemia cortex was assessed. Western blot was used to detect the expression of NF-κB in the ischemia cortex. The data were analyzed using SPSS 20.0 software, multiple-group comparisons were performed using one-way ANOVA, and LSD- t test was used for pairwise comparison between groups. A P<0.05 was considered statistically significant different. Results:Compared with the sham group, NDS, brain water content, relative infarction volume and rate of nerve cell necrosis were significantly increased in the I/R and Oxy groups (all P<0.05). Levels of TNF-α, IL-1β, IL-10, NF-κB and the activities of MPO were increased in the ischemia cortex (all P<0.05). Compared the Oxy group with the I/R group, NDS, brain water content, relative infarction volume and rate of nerve cell necrosis were significantly decreased [(1.7±0.9) vs (2.6±1.1);(79.2±2.4)% vs (84.7±4.2)%; (23.0±5.4)% vs (34.8±6.0)%; (25.2±12.4)% vs (54.8±14.8)%, all P<0.05]. The levels of TNF-α, IL-1β, relative expression of NF-κB, and the activities of MPO were significantly decreased in the ischemia cortex [(4.4±1.2) pg/mg vs (6.5±1.2) pg/mg; (5.4±0.7) pg/mg vs (7.8±0.8) pg/mg; (0.83±0.11) vs (1.23±0.33); (0.27±0.09) U/g vs (0.36±0.14) U/g, all P<0.05] , while the concentration of IL-10 was significantly increased [(20.9±4.5) pg/mg vs (9.2±1.6) pg/mg, t=6.036, P=0.000 1]. Conclusions:Oxycodone hydrochloride can attenuate focal cerebral I/R injury through inhibiting NF-κB activity.

Diabetes Mellitus and Cause-Specific Mortality: A Population-Based Study

BACKGROUND: To investigate whether diabetes contributes to mortality for major types of diseases. METHODS: Six National Health and Nutrition Examination Survey data cycles (1999 to 2000, 2001 to 2002, 2003 to 2004, 2005 to 2006, 2007 to 2008, and 2009 to 2010) and their linked mortality files were used. A population of 15,513 participants was included according to the availability of diabetes and mortality status. RESULTS: Participants with diabetes tended to have higher all-cause mortality and mortality due to cardiovascular disease, cancer, chronic lower respiratory diseases, cerebrovascular disease, influenza and pneumonia, and kidney disease. Confounder-adjusted Cox proportional hazard models showed that both diagnosed diabetes category (yes or no) and diabetes status (diabetes, prediabetes, or no diabetes) were associated with all-cause mortality and with mortality due to cardiovascular disease, chronic lower respiratory diseases, influenza and pneumonia, and kidney disease. No associations were found for cancer-, accidents-, or Alzheimer's disease-related mortality. CONCLUSION: The current study's findings provide epidemiological evidence that diagnosed diabetes at the baseline is associated with increased mortality risk due to cardiovascular disease, chronic lower respiratory diseases, influenza and pneumonia, and kidney disease, but not with cancer or Alzheimer's disease.