CXCR2基因敲除HeLa细胞株构建及转录组测序分析

目的通过基因敲除和转录组测序探究CXCR2相关功能基因及信号通路,揭示其在宫颈癌中的分子作用。方法设计构建靶向CXCR2基因的CRISPR-Cas9敲除质粒载体,转染HeLa细胞并筛选获得CXCR2基因敲除的细胞株。转录组测序分析差异表达基因,并进行基因本体(GO)分类富集分析和京都基因与基因组百科全书(KEGG)通路富集分析筛选信号通路和重要差异表达基因,差异表达基因经实时荧光定量PCR方法验证。结果成功构建了CXCR2基因敲除细胞株,转录组测序获得1519个差异表达基因,其中上调表达基因428个,下调表达基因1091个。GO富集分析显示了细胞组分、生物学过程、分子功能方面富集量前20的条目;KEGG信号通路富集分析显示,富集基因最多的为肿瘤相关通路,其次为PI3K-Akt信号通路和HPV感染信号通路。通过韦恩图分析获得了7个共同差异表达基因:EGF、FN1、ITGA2B、CCND3、PIK3CD、PDGFRB、CDKN1A,其中前6个基因表达显著下调,CDKN1A表达显著上调。结论转录组测序分析结果进一步提示CXCR2在宫颈癌中发挥作用,并初步确定了与其相关的信号通路和关键功能基因,为宫颈癌的分子机制和临床研究提供了理论基础。

Engineering human spinal microphysiological systems to model opioid-induced tolerance

pioids are commonly used for treating chronic pain.However,with continued use,they may induce tolerance and/or hyperalgesia,which limits therapeutic efficacy.The human mechanisms of opioid-induced tolerance and hyperalgesia are significantly understudied,in part,because current models cannot fully recapitulate human pathology.Here,we engineered novel human spinal microphysiological systems(MPSs)integrated with plug-and-play neural activity sensing for modeling human nociception and opioid-induced tolerance.Each spinal MPS consists of a flattened human spinal cord organoid derived from human stem cells and a 3D printed organoid holder device for plug-and-play neural activity measurement.We found that the flattened organoid design of MPSs not only reduces hypoxia and necrosis in the organoids,but also promotes their neuron maturation,neural activity,and functional development.We further demonstrated that prolonged opioid exposure resulted in neurochemical correlates of opioid tolerance and hyperalgesia,as measured by altered neural activity,and downregulation ofμ-opioid receptor expression of human spinal MPSs.The MPSs are scalable,cost-effective,easy-to-use,and compatible with commonly-used well-plates,thus allowing plug-and-play measurements of neural activity.We believe the MPSs hold a promising translational potential for studying human pain etiology,screening new treatments,and validating novel therapeutics for human pain medicine.

High mobility group box 1 levels as potential predictors of asthma severity

To the Editor:Asthma is an abnormal chronic inflammatory disease characterized by the involvement of a complex network of cells.Barrier epithelial cells(ECs)represent the first line of defense and express pattern recognition receptors to recognize type-2 cell-mediated immune insults.An exaggerated abnormal barrier function or inadequate immune response may contribute to the pathophysiology of asthma.

Fine particulate matter (PM2.5):The culprit for chronic lung diseases in China

Air pollution is a world public health problem. Particulate matter (PM), a mix of solid and liquid particles in the air, becomes an increasing concern in the social and economic development of China. For decades, epidemiological studies have confirmed the association between fine particle pollutants and respiratory diseases. It has been reported in different populations that increased fine particulate matter (PM2.5) concentrations cause elevated susceptibility to respiratory diseases, including acute respiratory distress, asthma, chronic obstructive pulmonary disease, and lung cancer. This review will discuss the pathophysiology of PM2.5 in res-piratory diseases, which are helpful for the prevention of air pollution and treatment of respiratory tract inflammatory diseases.

Association between socioeconomic status and chronic obstructive pulmonary disease in Jiangsu province, China: a population-based study

Background:Chronic obstructive pulmonary disease(COPD)is a common public health problem worldwide.Recent studies have reported that socioeconomic status(SES)is related to the incidence of COPD.This study aimed to investigate the association between SES and COPD among adults in Jiangsu province,China,and to determine the possible direct and indirect effects of SES on the morbidity of COPD.Methods:A cross-sectional study was conducted among adults aged 40 years and above between May and December of 2015 in Jiangsu province,China.Participants were selected using a multistage sampling approach.COPD,the outcome variable,was diagnosed by physicians based on spirometry,respiratory symptoms,and risk factors.Education,occupation,and monthly family average income(FAI)were used to separately indicate SES as the explanatory variable.Mixed-effects logistic regression models were introduced to calculate odds ratios(ORs)and 95%confidence intervals(CIs)for examining the SES-COPD relationship.A pathway analysis was conducted to further explore the pulmonary function impairment of patients with different SES.Results:The mean age of the 2421 participants was 56.63±9.62 years.The prevalence of COPD was 11.8%(95%CI:10.5%–13.1%)among the overall sample population.After adjustment for age,gender,residence,outdoor and indoor air pollution,body weight status,cigarette smoking,and potential study area-level clustering effects,educational attainment was negatively associated with COPD prevalence in men;white collars were at lower risk(OR:0.60,95%CI:0.43–0.83)of experiencing COPD than blue collars;compared with those within the lower FAI subgroup,participants in the upper(OR:0.68,95%CI:0.49–0.97)tertiles were less likely to experience COPD.Such negative associations between all these three SES indicators and COPD were significant among men only.Education,FAI,and occupation had direct or indirect effects on pulmonary function including post-bronchodilator forced expiratory volume in 1 s/forced vital capacity(FEV1/FVC),FEV1,FVC,and FEV1 percentage of predicted.Education,FAI,and occupation had indirect effects on pulmonary function indices of all participants mainly through smoking status,indoor air pollution,and outdoor air pollution.We also found that occupation could affect post-bronchodilator FEV1/FVC through body mass index.Conclusions:Education,occupation,and FAI had an adverse relationship with COPD prevalence in Jiangsu province,China.SES has both direct and indirect associations with pulmonary function impairment.SES is of great significance for COPD morbidity.It is important that population-based COPD prevention strategies should be tailored for people with different SES.

Air pollutants and early origins of respiratory diseases

Air pollution is a global health threat and causes millions of human deaths annually. The late onset of respiratory diseases in children and adults due to prenatal or perinatal exposure to air pollutants is emerging as a critical concern in human health. Pregnancy and fetal development stages are highly susceptible to environmental exposure and tend to develop a long-term impact in later life. In this review, we briefly glance at the direct impact of outdoor and indoor air pollutants on lung diseases and pregnancy disorders. We further focus on lung complications in later life with early exposure to air pollutants. Epidemiological evidence is provided to show the association of prenatal or perinatal exposure to air pollutants with various adverse birth outcomes, such as preterm birth, lower birth weight, and lung developmental defects, which further associate with respiratory diseases and reduced lung function in children and adults. Mechanistic evidence is also discussed to support that air pollutants impact various cellular and molecular targets at early life, which link to the pathogenesis and altered immune responses related to abnormal respiratory functions and lung diseases in later life.

Sialic acid-binding immunoglobulin-like lectin 9 as a potential therapeutic target for chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease(COPD)has become the third-leading cause of death worldwide,which is a severe economic burden to the healthcare system.Chronic bronchitis is the most common condition that contributes to COPD,both locally and systemically.Neutrophilic inflammation predominates in the COPD airway wall and lumen.Logically,repression of neutrophilia is an essential fashion to COPD treatment.However,currently available anti-neutrophilic therapies provide little benefit in COPD patients and may have serious side effects.Thus,there is an urgent need to explore an effective and safe anti-neutrophilic approach that might delay progression of the disease.Sialic acid-binding immunoglobulin-like lectin(Siglec)-9 is a member of the Siglec cell surface immunoglobulin family.It is noteworthy that Siglec-9 is highly expressed on human neutrophils and monocytes.Ligation of Siglec-9 by chemical compounds or synthetic ligands induced apoptosis and autophagic-like cell death in human neutrophils.Furthermore,administration of antibody to Siglec-E,mouse functional ortholog of Siglec-9,restrained recruitment and activation of neutrophils in mouse models of airway inflammation in vivo.Given the critical role that neutrophils play in chronic bronchitis and emphysema,targeting Siglec-9 could be beneficial for the treatment of COPD,asthma,fibrosis,and related chronic inflammatory lung diseases.

Histamine Excites Rat GABAergic Ventral Pallidum Neurons via Co-activation of H1 and H2 Receptors

The ventral pallidum(VP) is a crucial component of the limbic loop of the basal ganglia and participates in the regulation of reward, motivation, and emotion.Although the VP receives afferent inputs from the central histaminergic system, little is known about the effect of histamine on the VP and the underlying receptor mechanism. Here, we showed that histamine, a hypothalamicderived neuromodulator, directly depolarized and excited the GABAergic VP neurons which comprise a major cell type in the VP and are responsible for encoding cues of incentive salience and reward hedonics. Both postsynaptic histamine H1 and H2 receptors were found to be expressed in the GABAergic VP neurons and co-mediate the excitatory effect of histamine. These results suggested that the central histaminergic system may actively participate in VP-mediated motivational and emotional behaviors via direct modulation of the GABAergic VP neurons. Our findings also have implications for the role of histamine and the central histaminergic system in psychiatric disorders.

How will telomeric complex be further contributed to our longevity?—The potential novel biomarkers of telomere complex counteracting both aging and cancer

With the smooth move towards the coming expected clinical reports of anticancer pharmaceutical molecules targeting telomeres and telomerase,and also with the exciting success in the extension of lifespan by regulating telomerase activity without increased onset of oncogenesis in laboratory mouse models(Garcia-Cao et al.,2006;Jaskelioff et al.,2011),we are convinced that targeting telomeres based on telomerase will be a potential approach to conquer both aging and cancer and the idea of longevity seems to be no more mysterious.More interestingly,emerging evidences from clinical research reveal that other telomeric factors,like specifi c telomeric binding proteins and nonspecific telomere associated proteins also show crucial importance in aging and oncogenesis.This stems from their roles in the stability of telomere structure and in the inhibition of DNA damage response at telomeres.Uncapping these proteins from chromosome ends leads to dramatic telomere loss and telomere dysfunction which is more abrupt than those induced by telomerase inactivation.Abnormal expression of these factors results in developmental failure,aging and even oncogenesis evidenced by several experimental models and clinical cases,indicating telomere specifi c proteins and its associated proteins have complimentary roles to telomerase in telomere protection and controlling cellular fate.Thus,these telomeric factors might be potential clinical biomarkers for early detection or even therapeutic targets of aging and cancer.Future studies to elucidate how these proteins function in telomere protection might benefit patients suffering aging or cancer who are not sensitive to telomerase mediation.

The evaluation of reverse shoulder lateralization on deltoid forces and scapular fracture risk:A computational study

Reverse shoulder arthroplasty(RSA)can treat severe rotator cuff deficiency,but its medialized design of the shoulder's center of rotation(CoR)has been associated with scapular notching.Although lateralization of CoR provides a larger impingement-free range of motion,the changes in component positioning alter the biomechanics and may cause unforeseen complications.This work quantified the muscle forces and predicted the scapula fracture risk by coupling dynamic simulation analysis with finite element modeling.To identify bone failure patterns,the results were analyzed using three common failure measures.A parametric study showed greater lateralization produced higher strain/stress concentrations in the scapular spine(Levy Region Type II),with approximately a 10%increase for the 12 mm lateralized scenario,compared to a neutral configuration.Significant differences in fracture risk patterns were found between the maximum principal stress/strain results and the von Mises stress results.The lateralized configurations could increase the muscle and joint reaction forces during abduction and induce scapular fracture.Studying the effects of RSA lateralization on scapular fracture risk can help guide the continued optimization of RSA performance and surgical techniques.The findings of relationships between the loading style and bone failure measures can provide valuable insight into the investigation of bone failure criteria.