Objective To investigate the correlation between the levels of Clara cell secretory protein (CC16) and pulmonary surfactant protein A (SP-A) in cord blood serum and severe ICP-induced lung injury in preterm infants. Methods Totally 102 cord blood samples of 34 ～ 37 weeks premature infants were collected from the Third Affiliated Hospital of Chongqing Medical University from September 2019 to May 2021. They were divided into two groups: ICP group (56 cases) and control group (46 cases). The levels of CC16 and SP-A in cord blood serum were detected by enzyme-linked immunosorbent assay (ELISA). The incidence of lung injury and the concentrations of CCSP and SP-A in cord blood serum were compared between the two groups. Results The incidence of lung injury in group ICP was significantly higher than that in control group, and the levels of CC16 and SP-A in cord blood of group ICP were lower than those of control group (P ＜ 0.05). Conclusion The detection of CC16 and SP-A levels in cord blood can be used as a reference for early prediction and diagnosis of lung injury in severe ICP preterm infants.

Thanks to containing tumor‐related molecules, small extracellular vesicles (sEVs) are emerging as biomarkers in tumor liquid biopsy and commonly employed to diagnose a specific cancer. However, a pan‐cancer screening method for pre‐symptomatic patients is crucial to achieving the early cancer detection. Herein, a lipid‐protein capture system on herringbone (HB) microfluidic chip ( HB EV‐Chips) to isolate multiple tumor‐derived sEVs is constructed. Phosphatidylserine (PS) is abnormally surface‐supposed on tumor‐derived sEVs and binds T‐cell immunoglobulin domain and mucin domain‐containing protein 4 (Tim4) in calcium‐dependent manner. PS + sEVs isolated by the HB EV‐Chips is perform on liquid chromatography electrospray ionization tandem mass spectrometry and it is found that PS + sEVs are highly correlated with tumor‐related pathway ( P < 0.05), such as Cdc42 protein signal transduction, neuropilin signaling pathway, and Wnt signaling pathway. And it is further validated in clinical sample and found that PS + sEV number shows statistical difference between cancer patients and healthy donors in plasma ( P < 0.01) and has efficient diagnostic power (area under curve = 0.86), suggesting PS + sEV as potential biomarker for pan‐cancer screening. The Tim‐4 functionalized device facilitates the early multiple cancer detection, providing the potential to be used as a rapid‐screening tool in clinical setting.

Abstract Extracellular vesicles (EVs) of nanoscale carry the biological information of parent cells, however, there is no consensus on the culture, collection, and purification of cell-derived EVs. We optimized the concentration of extracelluar vesicle (EV)-depleted serum (EDS), time of collection, and the extraction method for small EVs (sEVs) derived from A549 cells to improve the yield and quality of sEVs. Firstly, A549 cells were cultured in media containing a concentration gradient of EDS (10%, 5%, 3%, 1%, 0.5%, and 0%) for a prolonged time to increase sEV yield. The effects of ultracentrifugation and total exosome isolation kit(EI-kit) on EV-specific proteins were compared, and the biological activity of sEVs was evaluated. There were no significant differences in cell morphology and viability over the range of 1%-10% of the EDS concentration (p > 0.05). The sEVs collected continuously for five days from the cells cultured with 3% EDS were 40-200nm in diameter and of double-membrane, spherical structure with expression of the specific proteins TSG101 and CD9. The sEVs on fifth day could be endocytosed by A549cell which suggested that the sEVs were biologically active.The results of LC-MS/MS demonstrated 3%-sEVs (i.e., sEVs from cells cultured with 3% EDS) have a comprehensive profiling of sEV proteome. In conclusion, 3% EDS is suitable for cell culture, and the cell culture supernatants can be collected continuously for 5 days while maintaining the biological activity of sEVs, and ultracentrifugation is superior to EI in extracting sEVs derived from A549 cells.

Pan-Cancer Screening In the background of the inside cover, highlighting article number 2101115 by Kun Deng and co-workers, the body is primarily made up of lung, colorectum, pancreas and blood vessels, which represents the concept of pan-cancer screening in tumor liquid biopsy. The drop of blood entrances into HBEV-Chip are indicating the isolation of tumor-derived small extracellular vesicles from microlitre volume plasma for pan-cancer screening in clinic and the high quality materials for mass spectrometry detection in lab.

Abstract Background Cervical squamous cell carcinoma (CSCC), caused by the infection of high‐risk human papillomavirus, is one of the most common malignancies in women worldwide. Methods RNA expression data, including those from the Cancer Genome Atlas, Gene Expression Omnibus, and Genotype‐Tissue Expression databases, were used to identify the expression of RNAs in normal and tumor tissue. Correlation analysis was performed to identify the immune‐related long noncoding RNAs (IRLs) and hypoxia‐related genes (IRHs) that can influence the activity of the immune system. Prognosis models of immune‐related RNAs (IRRs) were used to construct a coexpression network of the immune system. We identified the role of IRRs in immunotherapy by correlation analysis with immune checkpoint genes (ICGs). We then validated the expression data by integrating two single‐cell sequencing data sets of CSCC to identify the key immune features. Results In total, six immune‐related gene (IRG), four IRL, and five IRH signatures that can significantly influence the characteristics of the tumor immune microenvironment (TIME) were selected using machine learning methods. The expression level of ICGs was significantly upregulated in GZMB+CD8+ T‐cells and tumor‐associated macrophages (TAMs) in tumor tissues. TGFBI+ TAMs are a kind of blood‐derived monocyte‐derived M0‐like TAM linked to hypoxia and a poor prognosis. IFI30+ M1‐like TAMs participate in the process of immune‐regulation and showed a role in the promotion of CD8+ T‐cells and Type 1 T helper (Th1)/Th2 cells in the coexpression network, together with several IRLs, IRGs, and ICGs. Conclusions CD16+ monocyte‐derived IFI30+ TAMs participated in our coexpression network to regulate the TIME, showing the potential to be a novel immunotherapy target. The enrichment of M0‐like TAMs was associated with a worse prognosis in the high‐risk score group with IRH signatures. Remarkably, M0‐like TAMs in tumor tissues overexpressed TGFBI and were associated with several well‐known tumor‐proliferation pathways.

外泌体是一类参与多种生理病理过程的细胞间信息交流载体,其脂质双分子层结构能有效维护所携带的生物信息分子的稳定性.因此,外泌体相对于血清、血浆等更加适合于组学研究,如蛋白质组学、转录组学及代谢组学等.随着质谱技术的不断发展,蛋白质作为诸多生物功能的直接执行者受到越来越多的关注,尤其是外泌体内的蛋白质及其生物功能.外泌体用于蛋白质组学的研究有助于阐明肿瘤的发生和发展机制,进而寻找特异性的肿瘤标志物和可用于精准治疗的靶点.

Exosomes, which are lipid membrane-enclosed nanovesicles (30-150 nm in diameter), contain abundant proteomic and genetic information. Specifically, tumor-derived exosomes have been recognized as ideal biomarkers for cancer diagnosis. However, effective methods for isolating as well as detecting exosomes are still challenging. Herein, we have developed an immunoaffinity-based microfluidic device HBEXO-Chip which can directly separate exosomes from plasma. This separation technology enables capturing the exosomes through tumor specific surface markers, such as the pancreatic cancer target Glypican-1 employed in this study, to prevent the interference of non-specific exosomes. Compared with the traditional exosomes separation workflow, (EXO)-E-HB-Chip shows excellent specific exosomes separation performance. We demonstrated that GPC1+ exosomes can distinguish pancreatic cancer from Pancreatitis group and healthy control group. Additionally, we identificate a miRNA signature for pancreatic cancer through RNA sequencing. Finally, we found 2 miRNAs with diagnostic value for pancreatic cancer in these differentially expressed miRNAs, namely miR-125b-5p and miR-214-3p.

BACKGROUND:Hypercoagulation starts as early as the first-trimester pregnancy and is a risk factor for thromboembolic events which are associated with miscarriage. Our study aimed to investigate coagulation, platelets, and fibrinolysis parameters alteration amongst trimester-specific normal pregnancy and first-trimester miscarriage patients. We also test the accuracy of haemostatic parameters determination for prediction of first-trimester miscarriage.METHODS:Retrospective investigation of 50 women whose most recent pregnancy had ended in the first trimester and 54 age-matched consecutive normal pregnancy between 2016 and 2019. Furthermore, 51 non-pregnant, age-matched women were included in parallel to healthy controls. Twelve screening tests for coagulation and platelet parameters were assessed.RESULTS:We found plasma levels of aPTT, FBG, and TT were significantly prolonged or decreased in miscarriage subjects than the corresponding first phase in normal pregnancies. PT, INR, aPTT, and d-dimer all shift back to normal in miscarriage patients compared with non-pregnant women. Shortened aPTT combined with TT and FBG can predicted the occurrence of first-trimester miscarriage with an AUC of 0.831.CONCLUSIONS:Routine assessment of aPTT combined with TT and FBG is a low-cost, widely available marker for prediction of first-trimester miscarriage.

传统的医院教学和科研模式存在学科独立、知识结构单一、学科间交叉不足的缺点.这直接导致研究型医院处理疑难重症疾病的能力和科研创新动力不足.重庆医科大学附属教学医院引入中心制模式后, 学科间相互交融, 同时在探讨中引入先进的非医学学科理念, 如认知上提倡发展"团队医疗";技术上沟通合作如AI、纳米机器人和太赫兹等前沿技术.结合本科室的科研教学实践, 在以检验医师为出发点探讨重庆医科大学附属教学医院在当前中心制模式下教学、科研与学科建设状况, 为进一步改善检验医师融入医院中心制模式, 推动科研教学发展提供决策依据.

Methods based on potentiometric measurement have been developed for immunoassays, but most exhibit low sensitivities and are unsuitable for early diagnosis of disease. Herein we design a new potentiometric immunosensing platform for the sensitive detection of carcinoma antigen 15-3 (CA 15-3) by coupling with enzymatic biocatalytic precipitation and a nanogold labeling technique. The sandwich-type immunoreaction is carried out on a monoclonal anti-CA 15-3 capture antibody-modified working electrode, using horseradish peroxidase (HRP) and polyclonal anti-CA 15-3 detection antibody-labeled gold nanoparticles. Upon the introduction of target CA 15-3, the carried HRP molecules with an immunocomplex catalyze the oxidation of 4-chloro-1-naphthol (4-CN) into the insoluble benzo-4-chlorohexadienone. The formed product coated on the surface of the modified electrode results in a change of the electrical potential. Under optimal conditions, the shift in the electrical potential relative to the background signal increases with the increasing target CA 15-3 concentration, and exhibits a good linear relationship within the dynamic range of 0.01-30 U mL-1 at a detection limit of 7.8 mU mL-1. Good precision and reproducibility, and high specificity were acquired for the analysis of 15 human serum specimens, giving well matched results with those obtained from a human CA 15-3 ELISA kit.

A new homogeneous electrochemical sensing system was developed for sensitive detection of microRNA-21 (miRNA-21) based on target-induced glucose release from propylamine-functionalized mesoporous silica nanoparticle (MSN) with glucometer readout. Glucose molecules (as the signal tracers) were initially gated into the pores through the interaction of the negatively charged anchor DNA with the aminated MSN. Upon addition of target miRNA, the analyte competitively hybridized with anchor DNA to form the RNA-DNA duplex, thus resulting in detachment of anchor DNA from the MSN accompanying the pore opening. The loaded glucose molecules released out from the pores because of concentration gradients, which could be detected by using a portable personal glucometer (PGM). Experimental results indicated that the PGM signal increased with the increasing miRNA level, and exhibited a good linear dependence on the miRNA-21 concentration from 50 pM to 5.0 nM with a detection limit of 19 pM under optimum conditions. Additionally, multifunctional mesoporous silica nanoparticles also showed good stability and favorable selectivity, and satisfactory accuracy for the miRNA detection in cell lysates with quantitative real-time polymerase chain reaction (qRT-PCR). Such good analytical performance endows it as a promising scheme for the efficient and convenient detection of miRNA in clinical diagnosis and therapy.

Objective— Akkermansia muciniphila (A muciniphila) is a mucin-degrading bacterium that resides in the mucus layer whose abundance inversely correlates with body weight and the development of diabetes mellitus in mice and humans. The objective of this study was to explore the regulatory effect of A muciniphila on host lipoprotein metabolism, insulin sensitivity, and hepatic metabolic inflammation. Approach and Results— By establishing a novel mouse model that colonized the A muciniphila in the gastrointestinal tract of the cAMP-responsive binding protein H (CREBH)–deficient mouse and in vivo chylomicron assay, we found that increased colonization of A muciniphila in the gastrointestinal tract of wild-type mice protected mice from an acute fat load–induced hyperlipidemia compared with vehicle-treated mice. A muciniphila administration also significantly ameliorated chronic hypertriglyceridemia, improved insulin sensitivity, and prevented overproduction of postprandial chylomicrons in CREBH-null mice. Mechanistic studies revealed that increased A muciniphila colonization induced expression of low-density lipoprotein receptors and apolipoprotein E in the hepatocytes of CREBH-null mice, which facilitated the uptake of intermediate-density lipoprotein via the mediation of apolipoprotein B100 and apolipoprotein E, leading to the increased clearance of triglyceride-rich lipoprotein remnants, chylomicron remnants, and intermediate-density lipoproteins, from the circulation. Treatment with A muciniphila further improved hepatic endoplasmic reticulum stress and metabolic inflammation in CREBH-null mice. Conclusions— Increased colonization of the disease-protective gut bacteria A muciniphila protected the host from acute and chronic hyperlipidemia by enhancing the low-density lipoprotein receptor expression and alleviating hepatic endoplasmic reticulum stress and the inflammatory response in CREBH-null mice.

Akkermansia muciniphila ( A. muciniphila ) is a mucin-degrading bacterium that resides in the mucus layer whose abundance inversely correlates with body weight and the development of diabetes in mice and humans. The objective of this study was to explore the regulatory effect of A. muciniphila on host lipoprotein metabolism, insulin sensitivity and hepatic metabolic inflammation. By establishing a novel mouse model that colonized the A. muciniphila in the gastrointestinal(GI) tract of the cAMP-responsive binding protein H (CREBH)-deficient mouse and in vivo chylomicron assay, we found that increased colonization of A. muciniphila in the GI tract of wild-type mice protected mice from an acute fat load-induced hyperlipidemia compared to vehicle-treated mice. A. muciniphila administration also significantly ameliorated chronic hypertriglyceridemia, improved insulin sensitivity and prevented overproduction of postprandial chylomicrons in the CREBH-null mice. Mechanistic studies revealed that increased A. muciniphila colonization induced expression of low density lipoprotein (LDL) receptors and apolipoprotein E (apoE) in the hepatocytes of CREBH-null mice, which facilitated the uptake of intermediate density lipoprotein (IDL) via the mediation of apolipoprotein B100 (apoB) and apoE, leading to the increased clearance of triglyceride-rich very-low-density lipoprotein (VLDL) particles, the precursors of IDLs, from the circulation. Treatment with A. muciniphila further improved hepatic ER stress and metabolic inflammation in CREBH-null mice. Conclusion: Increased colonization of the disease-protective gut bacteria A. muciniphila protected the host from acute and chronic hyperlipidemia by enhancing the LDL receptor expression and alleviating hepatic ER stress and the inflammatory response in CREBH-null mice.

The activation of sterol regulatory element binding proteins (SREBPs) is regulated by insulin-induced genes 1 and 2 (Insig-1 and Insig-2) and SCAP. We previously reported that feeding R-α-lipoic acid (LA) to Zucker diabetic fatty (ZDF) rats improves severe hypertriglyceridemia. In this study, we investigated the role of cyclic AMP-responsive element binding protein H (CREBH) in the lipid-lowering mechanism of LA and its involvement in the SREBP-1c and Insig pathway. Incubation of McA cells with LA (0.2 mM) or glucose (6 mM) stimulated activation of CREBH. LA treatment further induced mRNA expression of Insig-1 and Insig-2a, but not Insig-2b, in glucose-treated cells. In vivo, feeding LA to obesity-induced hyperlipidemic ZDF rats activated hepatic CREBH and stimulated transcription and translation of Insig-1 and Insig-2a. Activation of CREBH and Insigs induced by LA suppressed processing of SREBP-1c precursor into nuclear SREBP-1c, which subsequently inhibited expression of genes involved in fatty acid synthesis, including FASN, ACC and SCD-1, and reduced triglyceride (TG) contents in both glucose-treated cells and ZDF rat livers. Additionally, LA treatment also decreased abundances of very low density lipoprotein (VLDL)-associated apolipoproteins, apoB100 and apoE, in glucose-treated cells and livers of ZDF rats, leading to decreased secretion of VLDL and improvement of hypertriglyceridemia. This study unveils a novel molecular mechanism whereby LA lowers TG via activation of hepatic CREBH and increased expression of Insig-1 and Insig-2a to inhibit de novo lipogenesis and VLDL secretion. These findings provide novel insight into the therapeutic potential of LA as an anti-hypertriglyceridemia dietary molecule.

Insulin resistant is commonly associated with both increased circulating levels of tumor necrosis factor‐alpha (TNF‐alpha) and hepatic overproduction of very low density lipoproteins (VLDL). The cAMP responsive binding protein H (CREBH) is a transcription factor whose expression is induced by proinflammatory cytokines. In this study, our objective was to investigate the role of CREBH in mediating lipogenic diet induced hyperlipoproteinemia in the insulin resistance rodent models. By using the wild type and CREBH‐null mouse models combined with fatty acid gavage and TNF‐alpha treatment, we demonstrated that depletion of CREBH reduced both mRNA and protein expression of apolipoprotein B (apoB), a key structure protein in VLDL particle. In vitro, transient expression of CREBH cDNAs in McA cells induced significant increase of apoB mRNA and protein expression and VLDL secretion which indicated the positive regulatory impact of CREBH on apoB biosynthesis. Furthermore, treating McA cells with TNF‐alpha activated CREBH and increased apoB protein level. Challenging the CREBH‐null mice with a dose of long chain fatty acid through gavage or TNF‐alpha treatment failed to stimulate apoB expression and the subsequent VLDL secretion which resulted in hepatic steatosis. Insulin treatment synergized the effect of CREBH on VLDL secretion which may be a contributing factor in insulin resistance induced hyperlipidemia. In conclusion, these data demonstrated that CREBH is the molecular link that mediates inflammatory cytokine signaling to overproduction of VLDL in hepatic insulin resistance. This novel finding provides new mechanistic insight in inflammatory cytokine induced hyperlipidemia in metabolic diseases

Activation of the sterol regulatory element-binding proteins (SREBPs), a step regulated by a cluster of ER-resident proteins, Insig-1, Insig-2 and SCAP, is rate limiting in hepatic de novo lipogenesis. We previously reported that feeding R-alpha-lipoic acid (LA) to ZDF (fa/fa) rats improves severe hypertriglyceridemia and lowers abdominal fat mass by inhibiting expression of genes involved in hepatic long-chain fatty acids and triacylglycerol syntheses. In this study, we characterized a novel mechanism of action of LA that explains its triacylglycerol lowering properties. Dietary LA activates liver specific transcription factor cAMP responsive element binding protein H (CREBH), which in turn enhances transcription and translation of Insig-1 and Insig-2. Chromatin immunoprecipitation (ChIP) assay demonstrated interaction between CREBH and the promoter of Insig-2 but not Insig-1. The increased abundance of Insig-1 and Insig-2 proteins contributes to sequester SREBP-1c and SREBP-2 in the ER and prevents their translocation to the Golgi apparatus where they would become activated. As a consequence, mRNA expression of genes involved in fatty acid and cholesterol synthesis, including FASN, ACC, SCD-1, HMGCR and LDL receptor, were significantly decreased in LA-fed animals versus pair-fed controls. Concomitantly, the assembly and secretion of very-low-density lipoproteins (VLDL) by primary hepatocytes were suppressed in the LA-fed ZDF rats as indicated by the decrease in VLDL-associated apolipoprotein B and apolipoprotein E. In vitro, treating a rat McA-RH7777 hepatoma cells with LA (200 micromole) activated CREBH, induced expression of Insig-1 and Insig-2, and hindered the palmitic acid-induced synthesis of triacylglycerol. This study provides new mechanistic insight into the triacylglycerol lowering properties of LA and supports the therapeutic potential of LA against hypertriglyceridemia.

Emerging evidence demonstrates a close interplay between disturbances in mitochondrial function and ER homeostasis in the development of the metabolic syndrome. The present investigation sought to advance our understanding of the communication between mitochondrial dysfunction and ER stress in the onset of hepatic steatosis in male rodents with defective peroxisome proliferator-activated receptor-α (PPARα) signaling. Genetic depletion of PPARα or perturbation of PPARα signaling by high-fructose diet compromised the functional activity of metabolic enzymes involved in mitochondrial fatty acid β-oxidation and induced hepatic mitochondrial stress in rats and mice. Inhibition of PPARα activity further enhanced the expression of apolipoprotein B (apoB) mRNA and protein, which was associated with reduced mRNA expression of the sarco/endoplasmic reticulum calcium ATPase (SERCA), the induction of hepatic ER stress, and hepatic steatosis. Restoration of PPARα activity recovered the metabolic function of the mitochondria and ER, alleviated systemic hypertriglyceridemia, and improved hepatic steatosis. These findings unveil novel roles for PPARα in mediating stress signals between hepatic subcellular stress-responding machinery and in the onset of hepatic steatosis under conditions of metabolic stress.

患者男,49岁,汉族,已婚,海南陵水黎族自治县农民,常年农耕.于2012年5月月 26日入院,主诉腹泻8d,腹胀4 d,意识不清伴低氧血症2h余.20年前因睾丸鞘膜积液行手术治疗,2个月前于外院诊断天疱疮.无其他病史和过敏史.患者自5月17日无明显诱因出现腹泻,黄色稀水样便,每次量约30～ 50 ml,10次/d,无发热、畏寒,无恶心、呕吐,无腹胀、腹泻、腹痛,无黑便及黏液脓血便;腹泻症状持续4 d无缓解.