Heyingwuzi formulation alleviates diabetic retinopathy by promoting mitophagy via the HIF-1α/BNIP3/NIX axis

BACKGROUND Diabetic retinopathy(DR)is the primary cause of visual problems in patients with diabetes.The Heyingwuzi formulation(HYWZF)is effective against DR.AIM To determine the HYWZF prevention mechanisms,especially those underlying mitophagy.METHODS Human retinal capillary endothelial cells(HRCECs)were treated with high glucose(hg),HYWZF serum,PX-478,or Mdivi-1 in vitro.Then,cell counting kit-8,transwell,and tube formation assays were used to evaluate HRCEC proliferation,invasion,and tube formation,respectively.Transmission electron microscopy was used to assess mitochondrial morphology,and Western blotting was used to determine the protein levels.Flow cytometry was used to assess cell apoptosis,reactive oxygen species(ROS)production,and mitochondrial membrane potential.Moreover,C57BL/6 mice were established in vivo using streptozotocin and treated with HYWZF for four weeks.Blood glucose levels and body weight were monitored continuously.Changes in retinal characteristics were evaluated using hematoxylin and eosin,tar violet,and periodic acid-Schiff staining.Protein levels in retinal tissues were determined via Western blotting,immunohistochemistry,and immunostaining.RESULTS HYWZF inhibited excessive ROS production,apoptosis,tube formation,and invasion in hg-induced HRCECs via mitochondrial autophagy in vitro.It increased the mRNA expression levels of BCL2-interacting protein 3(BNIP3),FUN14 domain-containing 1,BNIP3-like(BNIP3L,also known as NIX),PARKIN,PTEN-induced kinase 1,and hypoxia-inducible factor(HIF)-1α.Moreover,it downregulated the protein levels of vascular endothelial cell growth factor and increased the light chain 3-II/I ratio.However,PX-478 and Mdivi-1 reversed these effects.Additionally,PX-478 and Mdivi-1 rescued the effects of HYWZF by decreasing oxidative stress and apoptosis and increasing mitophagy.HYWZF intervention improved the symptoms of diabetes,tissue damage,number of acellular capillaries,and oxidative stress in vivo.Furthermore,in vivo experiments confirmed the results of in vitro experiments.CONCLUSION HYWZF alleviated DR and associated damage by promoting mitophagy via the HIF-1α/BNIP3/NIX axis.

Magnesium-L-threonate treats Alzheimer's disease by modulating the microbiota-gut-brain axis

Disturbances in the microbiota-gut-brain axis may contribute to the development of Alzheimer's disease. Magnesium-L-threonate has recently been found to have protective effects on learning and memory in aged and Alzheimer's disease model mice. However, the effects of magnesium-L-threonate on the gut microbiota in Alzheimer's disease remain unknown. Previously, we reported that magnesium-L-threonate treatment improved cognition and reduced oxidative stress and inflammation in a double-transgenic line of Alzheimer's disease model mice expressing the amyloid-β precursor protein and mutant human presenilin 1(APP/PS1). Here, we performed 16S r RNA amplicon sequencing and liquid chromatography-mass spectrometry to analyze changes in the microbiome and serum metabolome following magnesium-Lthreonate exposure in a similar mouse model. Magnesium-L-threonate modulated the abundance of three genera in the gut microbiota, decreasing Allobaculum and increasing Bifidobacterium and Turicibacter. We also found that differential metabolites in the magnesiumL-threonate-regulated serum were enriched in various pathways associated with neurodegenerative diseases. The western blotting detection on intestinal tight junction proteins(zona occludens 1, occludin, and claudin-5) showed that magnesium-L-threonate repaired the intestinal barrier dysfunction of APP/PS1 mice. These findings suggest that magnesium-L-threonate may reduce the clinical manifestations of Alzheimer's disease through the microbiota-gut-brain axis in model mice, providing an experimental basis for the clinical treatment of Alzheimer's disease.

Irisin/BDNF signaling in the muscle-brain axis and circadian system: A review

In mammals,the timing of physiological,biochemical and behavioral processes over a 24-h period is controlled by circadian rhythms.To entrain the master clock located in the suprachiasmatic nucleus of the hypothalamus to a precise 24-h rhythm,environmental zeitgebers are used by the circadian system.This is done primarily by signals from the retina via the retinohypothalamic tract,but other cues like exercise,feeding,temperature,anxiety,and social events have also been shown to act as non-photic zeitgebers.The recently identified myokine irisin is proposed to serve as an entraining non-photic signal of exercise.Irisin is a product of cleavage and modification from its precursor membrane fibronectin typeⅢdomain-containing protein 5(FNDC5)in response to exercise.Apart from well-known peripheral effects,such as inducing the"browning"of white adipocytes,irisin can penetrate the blood-brain barrier and display the effects on the brain.Experimental data suggest that FNDC5/irisin mediates the positive effects of physical activity on brain functions.In several brain areas,irisin induces the production of brain-derived neurotrophic factor(BDNF).In the master clock,a significant role in gating photic stimuli in the retinohypothalamic synapse for BDNF is suggested.However,the brain receptor for irisin remains unknown.In the current review,the interactions of physical activity and the irisin/BDNF axis with the circadian system are reconceptualized.

Brucea javanica oil emulsion improves the effect of radiotherapy on esophageal cancer cells by inhibiting cyclin D1-CDK4/6 axis

BACKGROUND Esophageal cancer is one of the most common cancers around the world, and it has high incidence and mortality rates. The conventional therapy for esophageal cancer is radiotherapy, although its effect is highly limited by the resistance of esophageal cancer cells. Thus, strong radiosensitizers can be very crucial during radiotherapy against esophageal cancer. Brucea javanica oil emulsion (BJOE) is a widely used drug against various cancers, such as liver, colon, and ovarian cancer. However, its anti-cancer effect and mechanism and the use of BJOE as a radiosensitizer have not been explored in esophageal cancer. AIM To evaluate the anti-cancer effect and mechanism of BJOE and explore the potential use of BJOE as a radiosensitizer during radiotherapy. METHODS The inhibitory effect of BJOE and its enhancement function with radiation on cell viability were examined with the calculated half-maximal effective concentration and half-maximal lethal concentration. The influence of BJOE on cell migration and invasion were measured with EC109 and JAR cells by wound-healing and transwell assay. Clonogenesis and apoptotic rate, which was measured by Hoechst staining, were investigated to confirm its enhancement function with radiation. To investigate the molecular pathway underlying the effect of BJOE, the expressions of several apoptosis- and cycle-related proteins was detected by western blotting.cell lines more than normal cell lines, and it markedly reduced migration and invasion in esophageal cancer cells (EC109 and JAR). Moreover, it promoted cell apoptosis and enhanced the effect of radiotherapy against esophageal cancerous cells. In the viability test, the values of half-maximal effective concentration and half-maximal lethal concentration were reduced. Compared to the control, only around 1/5 colonies formed when using BJOE and radiation together in the clonogenic assay. The apoptotic rate in EC109 was obviously promoted when BJOE was added during radiotherapy. Our study suggests that the expression of the apoptosis-proteins Bax and p21 were increased, while the expression of Bcl-2 was stable. Further detection of downstream proteins revealed that the expression of cyclin D1 and cyclin-dependent kinase 4/6 were significantly decreased. CONCLUSION BJOE has a strong anti-cancer effect on esophageal cancer and can be used as a radiosensitizer to promote apoptosis in cancerous esophageal cells via the cyclin D1-cyclin-dependent kinase 4/6 axis.

SOCS1/JAK2/STAT3 axis regulates early brain injury induced by subarachnoid hemorrhage via inflammatory responses

The SOCS1/JAK2/STAT3 axis is strongly associated with tumor growth and progression,and participates in cytokine secretion in many diseases.However,the effects of the SOCS1/JAK2/STAT3 axis in experimental subarachnoid hemorrhage remain to be studied.A subarachnoid hemorrhage model was established in rats by infusing autologous blood into the optic chiasm pool.Some rats were first treated with JAK2/STAT3 small interfering RNA(Si-JAK2/Si-STAT3)or overexpression plasmids of JAK2/STAT3.In the brains of subarachnoid hemorrhage model rats,the expression levels of both JAK2 and STAT3 were upregulated and the expression of SOCS1 was downregulated,reaching a peak at 48 hours after injury.Simultaneously,the interactions between JAK2 and SOCS1 were reduced.In contrast,the interactions between JAK2 and STAT3 were markedly enhanced.Si-JAK2 and Si-STAT3 treatment alleviated cortical neuronal cell apoptosis and necrosis,destruction of the blood-brain barrier,brain edema,and cognitive functional impairment after subarachnoid hemorrhage.This was accompanied by decreased phosphorylation of JAK2 and STAT3 protein,decreased total levels of JAK2 and STAT3 protein,and increased SOCS1 protein expression.However,overexpression of JAK2 and STAT3 exerted opposite effects,aggravating subarachnoid hemorrhage-induced early brain injury.Si-JAK2 and Si-STAT3 inhibited M1-type microglial conversion and the release of pro-inflammatory factors(inducible nitric oxide synthase,interleukin-1β,and tumor necrosis factor-α)and increased the release of anti-inflammatory factors(arginase-1,interleukin-10,and interleukin-4).Furthermore,primary neurons stimulated with oxyhemoglobin were used to simulate subarachnoid hemorrhage in vitro,and the JAK2 inhibitor AG490 was used as an intervention.The in vitro results also suggested that neuronal protection is mediated by the inhibition of JAK2 and STAT3 expression.Together,our findings indicate that the SOCS1/JAK2/STAT3 axis contributes to early brain injury after subarachnoid hemorrhage both in vitro and in vivo by inducing inflammatory responses.This study was approved by the Animal Ethics Committee of Anhui Medical University and the First Affiliated Hospital of University of Science and Technology of China(approval No.LLSC-20180202)on March 1,2018.

Effect of sericin on diabetic hippocampal growth hormone/insulin-like growth factor 1 axis

Previous studies have shown that sericin extracted from silk cocoon significantly reduces blood glucose levels and protects the nervous system against diabetes mellitus. In this study, a rat type 2 diabetes mellitus model was established by intraperitoneal injection of 25 mg/kg streptozotocin for 3 successive days, following which the rats were treated with sericin for 35 days. After treatment, the blood glucose levels of the diabetic rats decreased significantly, the growth hormone level in serum and its expression in the hippocampus decreased significantly, while the insulin-like growth factor-1 level in serum and insulin-like growth factor-1 and growth hormone receptor expression in the hippocampus increased significantly. The experimental findings indicate that sericin improves disorders of the growth hormone/insulin-like growth factor 1 axis to alleviate hippocampal damage in diabetic rats.

Localization of Porcine Neturopetide Y-Y1 mRNA in Hypothalamus-Pituitary-Ovary Axis during Puberty

[ Objective] To observe the disposition and localization of neturopetide Y-Y1 （ NPY-Y1 ） mRNA in hypothalamus, pituitary and ovary during puberty. [ Methed] Three 60-day-old Sujiang sows weighing 20 kg and three 160-day-old Sujiang sows weighing 80 kg were selected and anaesthetized. The hypothalamus, pituitary and ovary were taken out for preparation of frozen sections. The expression and localization of the NPY-Y1 mRNA in hypothalamus-pituitary-ovary axis was observed by the in situ hybridization. The PBS was substituted for hybridization solution to set up a control. ~ Result] Positive hybridization signals of the NPY-Y1 mRNA were detected in the hypothalamus, pituitary and ovary at different develop- mental stages, and these signals were stronger in the 60-day-old sows （before the puberty） than in the 160-day-old sows （during the puberty）. E Conclusion] The NPY-Y1 mRNA is distributed in the hypothalamus-pituitary-ovary axis and can regulate reproductive process of sows.

Metformin promotes anti-tumor immunity in STK11 mutant NSCLC through AXIN1-dependent upregulation of multiple nucleotide metabolites

Background:Metformin has pleiotropic effects beyond glucose reduction,including tumor inhibition and immune regulation.It enhanced the anti-tumor effects of programmed cell death protein 1(PD-1)inhibitors in serine/threonine kinase 11(STK11)mutant non-small cell lung cancer(NSCLC)through an axis inhibition protein 1(AXIN1)-dependent manner.However,the alterations of tumor metabolism and metabolites upon metformin administration remain unclear.Methods:We performed untargeted metabolomics using liquid chromatography(LC)-mass spectrometry(MS)/MS system and conducted cell experiments to verify the results of bioinformatics analysis.Results:According to the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway database,most metabolites were annotated into metabolism,including nucleotide metabolism.Next,the differentially expressed metabolites in H460(refers to H460 cells),H460_met(refers to metformin-treated H460 cells),and H460_KO_met(refers to metformin-treated Axin1-/-H460 cells)were distributed into six clusters based on expression patterns.The clusters with a reversed expression pattern upon metformin treatment were selected for further analysis.We screened out metabolic pathways through KEGG pathway enrichment analysis and found that multiple nucleotide metabolites enriched in this pathway were upregulated.Furthermore,these metabolites enhanced the cytotoxicity of activated T cells on H460 cells in vitro and can activate the stimulator of the interferon genes(STING)pathway independently of AXIN1.Conclusion:Relying on AXIN1,metformin upregulated multiple nucleotide metabolites which promoted STING signaling and the killing of activated T cells in STK11 mutant NSCLC,indicating a potential immunotherapeutic strategy for STK11 mutant NSCLC.

MYOD1 inhibits avian adipocyte differentiation via miRNA-206/KLF4 axis

Background:A considerable number of muscle development-related genes were differentially expressed in the early stage of avian adipocyte differentiation.However,the functions of them in adipocyte differentiation remain largely known.In this study,the myoblast determination protein 1(MYOD1)was selected as a representative of muscle development.We investigated its expression,function,and regulation in avian adipocyte differentiation.Results:The expression of MYOD1 decreased significantly in the early stage of avian adipocyte differentiation.CRIS PR/Cas9-mediated deletion of MYOD1 induced adipocyte differentiation,whereas over-expression of MYOD1 inhibited adipogenesis.The mRNA-seq data showed that MYOD1 could perturb the lipid biosynthetic process during differentiation.Our results showed that MYOD1 directly up-regulates the miR-206 expression by binding the upstream 1200 bp region of miR-206.Then,over-expression of miR-206 can inhibit the adipogenesis.Furthermore,MYOD1 affected the expression of endogenous miR-206 and its target gene Kruppel-like factor 4(KLF4),which is an important activator of adipogenesis.Accordingly,the inhibition of miR-206 or over-expression of KLF4 could counteract the inhibitory effect of MYOD1 on adipocyte differentiation.Conclusions:Our results establish that MYOD1 inhibits adipocyte differentiation by up-regulating miR-206 to suppress the KLF4 expression.These findings identify a novel function of MYOD1 in adipocyte differentiation,suggesting a potential role in body-fat distribution regulation.

Oridonin restores hepatic lipid homeostasis in an LXRa-ATGL/EPT1 axis-dependent manner

Hepatosteatosis is characterized by abnormal accumulation of triglycerides(TG),leading to prolonged and chronic inflammatory infiltration.To date,there is still a lack of effective and economical therapies for hepatosteatosis.Oridonin(ORI)is a major bioactive component extracted from the traditional Chinese medicinal herb Rabdosia rubescens.In this paper,we showed that ORI exerted significant protective effects against hepatic steatosis,inflammation and fibrosis,which was dependent on LXRa signaling.It is reported that LXRa regulated lipid homeostasis between triglyceride(TG)and phosphatidylethanolamine(PE)by promoting ATGL and EPT1 expression.Therefore,we implemented the lipidomic strategy and luciferase reporter assay to verify that ORI contributed to the homeostasis of lipids via the regulation of the ATGL gene associated with TG hydrolysis and the EPT1 gene related to PE synthesis in a LXRadependent manner,and the results showed the TG reduction and PE elevation.In detail,hepatic TG overload and lipotoxicity were reversed after ORI treatment by modulating the ATGL and EPT1 genes,respectively.Taken together,the data provide mechanistic insights to explain the bioactivity of ORI in attenuating TG accumulation and cytotoxicity and introduce exciting opportunities for developing novel natural activators of the LXRa-ATGL/EPT1 axis for pharmacologically treating hepatosteatosis and metabolic disorders.

Longitudinal assessment of peripheral organ metabolism and the gut microbiota in an APP/PS1 transgenic mouse model of Alzheimer’s disease

Alzheimer’s disease not only affects the brain,but also induces metabolic dysfunction in peripheral organs and alters the gut microbiota.The aim of this study was to investigate systemic changes that occur in Alzheimer’s disease,in particular the association between changes in peripheral organ metabolism,changes in gut microbial composition,and Alzheimer’s disease development.To do this,we analyzed peripheral organ metabolism and the gut microbiota in amyloid precursor protein-presenilin 1(APP/PS1)transgenic and control mice at 3,6,9,and 12 months of age.Twelve-month-old APP/PS1 mice exhibited cognitive impairment,Alzheimer’s disease-related brain changes,distinctive metabolic disturbances in peripheral organs and fecal samples(as detected by untargeted metabolomics sequencing),and substantial changes in gut microbial composition compared with younger APP/PS1 mice.Notably,a strong correlation emerged between the gut microbiota and kidney metabolism in APP/PS1 mice.These findings suggest that alterations in peripheral organ metabolism and the gut microbiota are closely related to Alzheimer’s disease development,indicating potential new directions for therapeutic strategies.

Sleeve gastrectomy ameliorates endothelial function and prevents lung cancer by normalizing endothelin-1 axis in obese and diabetic rats

BACKGROUND Previous evidence has implied that obesity is an independent risk factor for developing cancer.Being closely related to obesity,type 2 diabetes mellitus provides a suitable environment for the formation and metastasis of tumors through multiple pathways.Although bariatric surgeries are effective in preventing and lowering the risk of various types of cancer,the underlying mechanisms of this effect are not clearly elucidated.AIM To uncover the role and effect of sleeve gastrectomy(SG)in preventing lung cancer in obese and diabetic rats.METHODS SG was performed on obese and diabetic Wistar rats,and the postoperative transcriptional and translational alterations of the endothelin-1(ET-1)axis in the lungs were compared to sham-operated obese and diabetic rats and age-matched healthy controls to assess the improvements in endothelial function and risk of developing lung cancer at the postoperative 4 th,8 th,and 12 th weeks.The risk wasalso evaluated using nuclear phosphorylation of H2 A histone family member X as a marker of DNA damage(double-strand break).RESULTS Compared to obese and diabetic sham-operated rats,SG brought a significant reduction to body weight,food intake,and fasting blood glucose while improving oral glucose tolerance and insulin sensitivity.In addition,ameliorated levels of gene and protein expression in the ET-1 axis as well as reduced DNA damage indicated improved endothelial function and a lower risk of developing lung cancer after the surgery.CONCLUSION Apart from eliminating metabolic disorders,SG improves endothelial function and plays a protective role in preventing lung cancer via normalized ET-1 axis and reduced DNA damage.

Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(银杏蜜环口服溶液) attenuates inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis in post stroke depression

OBJECTIVE The Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(Yinxingmihuan Koufu Rongye,YXMH),a representative drug for"Treating both Brain and Heart",showed considerable clinical effects in isch⁃emic cardiovascular and cerebral vascular diseases.Recently,it is reported that YXMH has the potential for treating myocardial and cerebral ischemia related mental disorders,such as post stroke depression(PSD)and chronic heart disease(CHD)associated anxiety disorder.However,its mechanism has not been clearly elucidated.Meanwhile,increasing evidence revealed that there are close functional links between depression and habenular nucleus.The present study investigates the underlying mechanism of YXMH on attenuating the inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis in in a rat model of PSD.METHODS Rats were randomly devided into sham group,model group,Ginaton group(18 mg·kg^-1),Armillariella Mellea group(600 mg·kg-1),Fluoxetine group(10 mg·kg^-1),YXMH high-dose group(618 mg·kg^-1)and YXMH low-dose group(309 mg·kg^-1).The PSD model was induced by transarterial microembolization combined with sleep deprivation(2-Chloro-D-phenylalanine,PCPA,IH,200 mg·kg^-1,for 3 times,before the behavior test)in SD male rats.Then rats were treated with corresponding medicaments through gavage once a day until 3 weeks later,followed by body mass measurement,neurological deficit score evaluation,gripping strength and thermal withdrawl latency measurement,as well as depression related behavioral indicators,the open field test(OFT)and sucrose preference test.The pathological morphological changes of habenular nucleus was observed by HE staining,the expression of IBA-1 was measured and analyzed by immunohistochemistry staining,and alterations of proteins and genes related to the CX3CL1-CX3CR1 axis were analyzed using Western blotting(CX3CL1,CX3CR1)and real-time polymerase chain reaction(PCR)(CX3CL1,CX3CR1).RESULTS Compared with the sham group,rats in the model group manifested as decreased body mass,deficient neurological behavior and gripping strength,reduced loco⁃motor activity and sugar water consumption,as well as elevated thermal withdrawl latency(P<0.05,P<0.01).Mean⁃while,the pathological morphology of the habenular nucleus on the ischemic hemisphere showed significant neuronal degeneration,microglial proliferation,inflammatory cells and glia cells infiltration,together with up-regualted expression of IBA-1,CX3CL1,CX3CR1 protein and CX3CL1,CX3CR1 mRNA.YXMH attenuated inflammation of microglia in habenular nucleus through improving pathological morphology,inhibiting IBA-1 activation,down-regulating the expres⁃sion of CX3CL1 and CX3CR1 proteins and genes,and thus improved the behavior performance of ischemic injury and depression.CONCLUSION YXMH ameliorates neurological deficit and depressive behavior in rat model of PSD induced by transarterial microembolization combined with sleep deprivation,and the mechanism is probably related to attenu⁃ating inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis.

Analysis of feasibility of application of Beishu point acupressure therapy in treating gastroesophageal reflux disease via AMPK / ULK1 mediated autophagy pathway based on "adjusting central axis via pivot" theory

Gastroesophageal reflux disease(GERD)is a digestive system disease characterized by uncomfortable symptoms caused by reflux of gastric contents.It has increased sharply with the development of my country’s society and economy.If there is no reasonable and effective Prevention and treatment measures will inevitably increase the financial burden of patients,and also pose a major threat to the quality of life and health of patients.Cell signal transduction mediated by various receptors participates in the regulation mechanism of the body's various levels of biological functions.By inhibiting or activating its functions,the purpose of curing diseases can be achieved,and cell signal transduction has been used in traditional Chinese medicine.Studying.The theory of"adjusting the central axis"was explored by Professor Xie Sheng through decades of clinical experience.It has been proven in practice to treat GERD.It starts from the model of TCM viscera and expounds that the pathogenesis of GERD involves multiple viscera.Multi-system and multi-factor,explain the correlation of the disease with a variety of zang-fu syndromes,and use this as a basis to guide the clinical use of hidden prescriptions.The back-shu pointer therapy can prevent GERD by correcting the unbalanced state of the viscera and qi machine,and promoting the junction of the two channels of Ren and Du.Based on the theory of"adjusting the hub by the pivot",this article expounds the pathogenesis of GERD from the perspective of traditional Chinese medicine.By consulting the literature and combining with the previous research,it proposes to analyze the methods and methods of Backshu pointer therapy to prevent and treat GERD from the AMPK/ULK1 mediated autophagy pathway.

SDC1 acts as a novel molecular marker and prevents rheumatoid arthritis and modulates inflammatory response by targeting the miR-4531/SDC1 axis

Objective:Rheumatoid arthritis(RA)is a systemic autoimmune disease characterized by chronic erosive arthritis.Due to the lack of effective biomarkers for diagnosis and treatment,RA patients have many complications in the later stage,seriously affecting their quality of life.Thus,this study was conducted to investigate new therapeutic targets and to discover diagnostic biomarkers in RA.Methods:In this study,the expression profiles of GSE55235 and GSE55457 were downloaded from the Gene Expression Omnibus database to obtain DEGs between RA and healthy samples.Genetic Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed on the common genes existing in the RA-related modules.Additionally,we used the STRING database to construct the protein‒protein interaction network.Furthermore,we established the interaction analysis of Hub Genes and microRNA(miRNA)and verified the 10 Hub genes through the GSE77298 dataset and quantitative real-time polymerase chain reaction Results:276 and 69 DEGs were screened from the GSE55235 dataset and GSE55457 dataset,respectively.Then,we obtained 42 up-regulated genes in two chip datasets intersection.Genetic Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis of the 42 up-regulated genes showed that they were mainly concentrated in immune response-activating cell surface receptor signaling pathway,etc.Furthermore,the protein-protein interaction network indicated that 10 hub genes are closely related to RA,including MS4A1,CD27,LCK,CD79A,SDC1,CXCL9,CXCL10,CXCL13,IGLL5,and IGJ.In addition,we found that miR-4531 is the same target miRNAs between MS4A1 and SDC1 through messenger RNA-miRNA co-expression network.Finally,the GSE77298 gene chip and quantitative real-time polymerase chain reaction verified the expression of 10 Hub genes.The six Hub genes of CD27,SDC1,CXCL9,CXCL10,CXCL13,and IGJ are significantly increased.Conclusions:We found that SDC1 may be a novel molecular marker for the prevention and treatment of RA.The miR-4531/SDC1 regulatory axis may play a key role in this process.In conclusion,our study not only provides potential biomarkers for the diagnosis and treatment of RA,but also provides a basis and new targets for further revealing the potential mechanism of RA occurrence and development and discovering targeted drugs.

EPO在缺铁性贫血大鼠GH-IGF-1轴中的作用研究

目的探析EPO在缺铁性贫血大鼠GH-IGF-1轴中的作用价值。方法选取50只清洁级SD大鼠,均为雌性,鼠龄为3周龄,体重为60-65 g,通过特制饲料来制备缺铁性贫血大鼠模型,饲养7 d后按照体重随机分组,将其分为常规组(25例)与观察组(25例),给予常规组大鼠生理盐水,给予观察组大鼠重组人工红细胞生成素注射液,ELISA试验检查大鼠血清中的GH、IGF-1含量。结果干预前,两组大鼠血清GH水平无明显差异,差异无统计学意义(P>0.05);干预后,观察组大鼠血清GH水平明显低于常规组,差异有统计学意义(P<0.05);干预前,两组大鼠血清IGF-1水平无明显差异,差异无统计学意义(P>0.05);干预后,观察组大鼠血清IGF-1水平明显低于常规组,差异有统计学意义(P<0.05)。结论缺铁性贫血个体中体重下降、生长缓慢与血清中的GH、IGF-1水平有关,缺铁抑制了GH-IGF-1轴,对大鼠的生长发育造成影响,而将EPO升高能够对GH、IGF-1进行调节,对体重方面的控制有着积极意义。

运用CRISPR/Cas9技术构建敲除体轴抑制蛋白1(AXIN1)基因的ACT-1人未分化甲状腺癌细胞系

目的构建体轴抑制蛋白1(AXIN1)基因敲除的ACT-1人未分化甲状腺癌单克隆细胞系。方法用分子克隆技术及成簇的规律间隔的短回文重复序列/Cas9核酸酶(CRISPR/Cas9)基因编辑技术构建AXIN1基因敲除的单克隆细胞系;采用实时荧光定量PCR检测ACT-1细胞的AXIN1 mRNA水平,Western blot法检测AXIN1蛋白水平。结果通过T7检测成功获得有效的两个单链导向RNA(sgRNA)Cr3及Cr5;通过酶切鉴定及测序成功构建携带绿色荧光蛋白(GFP)的靶向AXIN1的sgRNA病毒载体;成功获得包括阴性对照在内的4个单克隆ACT-1未分化甲状腺癌细胞系;敲除组细胞AXIN1 mRNA及蛋白水平均明显降低。结论采用CRISPR-Cas9技术成功建立了敲除AXIN1基因的ACT-1未分化甲状腺癌细胞系。

木犀草素对神经性疼痛模型大鼠MCP-1/CCR2信号轴的影响

目的:探讨木犀草素调节单核细胞趋化蛋白-1(MCP-1)/CC趋化因子受体2(CCR2)信号轴,对神经性疼痛(NP)模型大鼠神经胶质细胞激活和免疫炎症的影响。方法:采用坐骨神经慢性压迫损伤法构建大鼠NP模型。将大鼠按随机数字表法分为模型组、阿魏酸钠组及木犀草素低、中、高剂量组,每组12只;另选择同期12只大鼠为假手术组。各组大鼠腹腔注射及灌胃相应药物,每天1次,连续14 d。测定大鼠机械性缩足反射阈值(MWT)和热刺激缩足反射潜伏期(TWL);实时荧光定量聚合酶链反应(qRT-PCR)及免疫组织化学法检测脊髓中胶质纤维酸性蛋白(GFAP)及小胶质细胞标志物离子钙接头蛋白分子1(Iba-1)mRNA及蛋白表达;流式细胞仪检测大鼠外周血中CD4^(+)、CD8^(+)水平;苏木素-伊红(HE)染色观察大鼠脊髓病理损伤情况;酶联免疫吸附试验(ELISA)检测脊髓中炎症因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子(TNF-α)水平;蛋白质印迹法(Western blotting)检测脊髓中MCP-1、CCR2蛋白表达。结果:与假手术组比较,模型组大鼠MWT、TWL、CD4^(+)T细胞比例及CD4^(+)/CD8^(+)比值降低,Iba-1、GFAP mRNA及蛋白表达、CD8^(+)T细胞比例、炎症因子水平(IL-6、IL-1β、TNF-α)、MCP-1及CCR2蛋白表达升高(P<0.05);与模型组比较,木犀草素低、中、高剂量组及阿魏酸钠组大鼠MWT、TWL、CD4^(+)T细胞比例及CD4^(+)/CD8^(+)比值升高,Iba-1、GFAP mRNA及蛋白表达、CD8^(+)T细胞比例、炎症因子水平(IL-6、IL-1β、TNF-α)、MCP-1及CCR2蛋白表达降低,且木犀草素作用效果呈剂量依赖性(P<0.05)。结论:木犀草素具有抗炎、增强免疫、抑制胶质细胞活化,缓解NP的作用,其作用机制可能与抑制MCP-1/CCR2信号轴的激活有关。

SINGULAR INTEGRAL EQUATIONS ON THE REAL AXIS WITH SOLUTIONS HAVING SINGULARITIES OF HIGHER ORDER

We transform the singular integral equations with solutions simultaneously having singularities of higher order at infinite point and at several finite points on the real axis into ones along a closed contour with solutions having singularities of higher order, and for the former obtain the extended Neother theorem of complete equation as well as the solutions and the solvable conditions of characteristic equation from the latter. The conclusions drawn by this article contain special cases discussed before.