Insulin resistance is often characterized as the factor that contributes to the emergence of metabolic diseases. Hepatic microRNAs (miRNAs) played critical roles in the development of metabolic-associated steatotic liver disease (MASLD) and insulin resistance. To investigate the effects of hepatic miR-411-5p in regulating insulin resistance, the present study utilized primary mouse hepatocytes and mice with MASLD. Suppression of miR-411-5p decreased hepatocyte glycogen production and phosphorylation of AKT, but miR-411-5p mimic improved insulin sensitivity. Mechanistically, 3'-UTR of transcription factor Sp2 was one of the binding sites of miR-411-5p. Treatment of miR-411-5p mimic suppressed the Sp2 mRNA and protein levels, enhancing the insulin signaling activity in the primary mouse hepatocytes. Hepatocyte-specific overexpression of Sp2 induced hepatic lipid accumulation and activation of related metabolic pathways. In contrast, inhibition of miR-411-5p reversely upregulated the expression of Sp2 and exaggerated insulin resistance in primary hepatocytes and the mouse model. Similarly, miR-411-5p mimic decreased obesity-induced hyperinsulinemia, glucose intolerance, insulin intolerance, and pyruvate intolerance. Furthermore, the parameters of MASLD, including lipid deposits, inflammation, and fibrosis, were improved after miR-411-5p replenishment, but co-administration with adeno-associated virus (AAV)-Sp2 abolished these benefits in the obese mouse model. Taken together, these findings demonstrated that Sp2-dependent miR-411-5p action regulates insulin resistance and MASLD, which provides a therapeutic approach toward resolving insulin resistance.

Non-typeable Haemophilus influenzae (NTHi) is a common pathogen causing respiratory infections, including pneumonia in children, and can also be found in the upper respiratory tracts of asymptomatic individuals. This study examines genomic variations between NTHi strains from healthy children and those from children with acute or chronic community-acquired pneumonia (CAP). Using bacterial genome-wide association studies (bGWAS), we compared these strains to identify key differences. Our analysis revealed that approximately 32% of genes exhibit variations between commensal and pathogenic states. Notably, we identified changes in peptidoglycan biosynthesis pathways and significant virulence factors associated with pneumonia. Furthermore, we observed a significant difference in β-lactam resistance due to PBP3 mutations between the healthy and pneumonia groups, confirmed by the ampicillin susceptibility test and characterized by the mutation pattern D350N, S357N, S385T, L389F. These findings contribute valuable insights into the genomic basis of NTHi pathogenicity and may inform more targeted clinical diagnostics and treatments.

BACKGROUND:The aim of the study was to conduct a comprehensive performance evaluation of the Mindray CX-9000 fully automated coagulation analyzer for the detection of the seven coagulation items. METHODS:The performance of activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), thrombin time (TT), D-dimer (D-D), fibrinogen degradation product (FDP), and antithrombin Ⅲ (AT) was validated for precision, linear range, carryover contamination rate, reference interval validation, inter-method agreement, and anti-interference ability. RESULTS:The intra- and inter-precision (coefficient of variation, CV%) of all seven items was less than the target CV%; the carryover contamination rates for different concentrations and between items were < 10%. The slope of the linear regression equation for the theoretical and measured values of the linear range was within 1 ± 0.05 and R ≥ 0.975. The reference interval quoted from the manufacturer's reference interval passed ≥ 95%. The CX-9000 was compared with the results of the reference instrument STAGO R MAX (STA-R MAX) and the p-values for all items ranged from 0.822 to 0.987. Within the concentration range claimed by the manufacturer, the interference errors produced by all items met the manufacturer's claimed criteria, except for triglycerides which produced interference errors > 10% for the FIB, D-D, FDP, and bilirubin which produced interference errors for the FIB and D-D assays. CONCLUSIONS:The CX-9000 automatic coagulation analyzer has good stability and repeatability, a wide linear range of detection, low carryover contamination rate, and high resistance to interference, making it suitable for the testing of clinical specimens.

Abstract Sepsis is a systemic inflammatory response syndrome caused by the invasion of pathogenic microorganisms such as bacteria into the body. PANoptosis is an inflammatory programmed cell death with key characteristics of pyroptosis, apoptosis, and/or neoptosis. At present, there is no strong evidence to suggest that the prognosis of sepsis is closely related to PANoptosis. In this study, 38 key differentially expressed genes(DEGs) were obtained by analyzing DEGs in sepsis microarray data GSE65685 and GSE95233 and crossing them with the PANopotosis gene set. Then, gene features were screened through univariate analysis, lasso regression analysis, and multivariate COX regression analysis to construct a prognosis model consisting of three predictive features: IKBKB, AIM2, and CTSG. We used Kaplan Meier (K-M) survival analysis, receiver operating characteristic (ROC) time curve analysis, internal validation, and principal component analysis to evaluate the performance of the prognostic model. In addition, sepsis patients were divided into high-risk and low-risk groups based on risk scores and gene set enrichment analysis (GSEA) results, and significant differences were found in multiple immune cell functions and immune related KEGG signaling pathways. Subsequently, scRNA seq data and immune cell infiltration analysis showed that the IKBKB and AMI2 genes were highly expressed in all immune cells of sepsis patients, while the CTSG gene was mainly highly expressed in monocytes, neutrophils, NK cells, and proliferative T cells. In the analysis of target genes for 16 immunosuppressive drugs, only CSF3 was highly expressed in high-risk patients, indicating that CSF3 may be the most promising target for treating sepsis.

脓肿分枝杆菌极少引起正常人群感染,但诊断和治疗较难.该菌可引起创伤、文身、皮外伤、外科手术后的皮肤感染[1],在免疫功能抑制人群中可能引起肺部感染和全身性感染[2].由于该菌对多种抗生素耐药,感染的诊断和治疗常依赖病原学检查结果.

Emerging evidence suggests that atherosclerosis, one of the leading phenotypes of cardiovascular diseases, is a chronic inflammatory disease. During the atherosclerotic process, immune cells play critical roles in vascular inflammation and plaque formation. Meanwhile, gastrointestinal disorder is considered a risk factor in mediating the atherosclerotic process. The present study aimed to utilize sivelestat, a selective inhibitor of neutrophil elastase, to investigate its pharmacological benefits on atherosclerosis and disclose the gastrointestinal–vascular interaction. The activation of intestinal neutrophil was increased during atherosclerotic development in Western diet-fed ApoE-/- mice. Administration of sivelestat attenuated atherosclerotic phenotypes, including decreasing toxic lipid accumulation, vascular monocyte infiltration, and inflammatory cytokines. Sivelestat decreased intestinal permeability and endotoxemia in atherosclerotic mice. Mechanistically, sivelestat upregulated the expression of zonula occludens-1 in the atherosclerotic mice and recombinant neutrophil elastase protein-treated intestinal epithelial cells. Meanwhile, treatment of sivelestat suppressed the intestinal expression of inflammatory cytokines and NF-κB activity. In contrast, administration of lipopolysaccharides abolished the anti-atherosclerotic benefits of sivelestat in the Western diet-fed ApoE-/- mice. Further clinical correlation study showed that the circulating endotoxin level and intestinal neutrophil elastase activity were positively correlated with carotid intima-medial thickness in recruited subjects. In conclusion, sivelestat had pharmacological applications in protection against atherosclerosis, and intestinal homeostasis played one of the critical roles in atherosclerotic development.

目的:探究肝纤维化标志物和HBV-DNA联合检测在慢性HBV感染后疾病进程中的应用价值.方法:回顾性分析2017年1月-2019年12月诊断为HBV携带者58例(对照组)、慢性乙型肝炎患者107例(慢性乙型肝炎组)、乙肝肝硬化患者34例(肝硬化组)的病历资料,检测并观察三组的肝纤维化标志物HA、PCⅢ、Ⅳ-C、LN和HBV-DNA载量,分析各指标之间及各指标与疾病进程的相关性,并绘制受试者工作特征(ROC)曲线.结果:肝纤维化四项指标HA、PCⅢ、Ⅳ-C、LN在对照组、慢性乙型肝炎组、肝硬化组中的含量依次增加,HBV-DNA载量在慢性乙型肝炎组中最高,其次是对照组;各指标在各组中的检测结果比较,差异均有统计学意义(P<0.05).血清肝纤维化四项指标HA、PCⅢ、Ⅳ-C、LN之间均呈正相关关系(P<0.01),其中PCⅢ与Ⅳ-C之间为高度相关(rs=0.897,P<0.01).血清肝纤维化四项指标HA、PCⅢ、Ⅳ-C、LN与HBV感染后疾病发展呈正相关(P<0.05),其中HA与HBV感染后疾病进程显著相关(rs=0.548,P<0.01).HA、PCⅢ、Ⅳ-C、LN、HBV-DNA单一检测对应AUC分别为0.925、0.929、0.916、0.899、0.630,联合检测对应AUC为0.959.结论:HBV在慢性乙型肝炎患者中呈现较高水平的复制,在肝硬化阶段复制水平降低.肝纤维化四项指标在HBV感染后随着疾病发展含量升高,其含量与疾病发展呈正相关关系,HA、PCⅢ、Ⅳ-C、LN四项指标之间也具有正相关关系,其中PCⅢ与Ⅳ-C含量高度相关,HA含量与疾病进程明显相关.联合检测肝纤维化四项血清学标志物与HBV-DNA载量可提高肝纤维化的诊断效能.

ObjectiveTo investigate the effect of hepatitis B virus (HBV) infection on the activation of hepatic stellate cells (HSCs) and its mechanism of action. MethodsA total of 30 plasma samples of chronic hepatitis B patients, 42 plasma samples of hepatitis B cirrhosis patients, 30 plasma samples of hepatocellular carcinoma patients, and 18 plasma samples of the individuals undergoing physical examination were collected from November 2020 to January 2021, and ELISA was used to measure the content of hepatitis B X protein (HBx), transforming growth factor-β1 (TGFβ1), dopamine beta-hydroxylase (DBH), and hydroxyproline (HYP) in plasma and conditioned medium. LO2 cells were used to establish a cell line with stable overexpression of HBx (LO2-HBx) and negative control cells (LO2-con), and a conditioned medium was prepared for LO2-HBx, LO2-Con, and LO2 cells (Mock), respectively; human HSC cell line LX-2 was incubated and divided into LX-2/LO2-HBx, LX-2/LO2-con, and LX-2/Mock groups, and CCK-8 assay was used to measure the change in cell proliferation. LX-2 cells were stimulated by rhTGFβ1, and the cells in the LX-2/LO2-HBx group were treated with a TGFβ1 receptor inhibitor. Quantitative real-time PCR and Western blot were used to measure the expression of HBx in LO2 cells and the expression of alpha-smooth muscle actin (α-SMA), collagen type I alpha 1 (Col1A1), DBH, and TGFβ1 in the above LX-2 cells. An analysis of variance was used for comparison between multiple groups, and the Bonferroni method was used for further comparison; the t-test was used for comparison between two groups; the Pearson method was used for correlation analysis. ResultsLO2-HBx stably expressed HBx protein and showed an increase in the content of TGFβ1 in supernatant (F=324.701, P＜0.01). The co-cultured LX-2/LO2-HBx group had a significant change in cell morphology, with the presence of cell shrinkage, extended cytoplasmic process, and reduced lipid droplets, and compared with the LX-2/LO2-con group, the LX-2/LO2-HBx group had significant increases in proliferative activity (P＜005) and the mRNA and protein expression levels of α-SMA and Col1A1 (mRNA: F=144.712 and 76.680, both P＜001; protein: F=234.142 and 528.708, both P＜0.001). The LX-2/LO2-HBx group had significant increases in the content of TGFβ1 (F=29.382, P＜001) and DBH (F=42.662, P＜0.01). With the increase in the stimulating concentration of rhTGFβ1, there were significant increases in the expression of α-SMA (F=1 794.031, P＜0.01), Col1A1 (F=91.340, P＜0.01), and DBH (F=2 501.011, P＜0.01), which reached the peak values at the rhTGFβ1 concentration of 10 ng/ml, and after a TGFβ1 receptor inhibitor was added to the conditioned medium, the LO2-HBx group had significant reductions in the expression of DBH and Col1A1 compared with the control group (t=3.603 and 5798, both P＜0.05). Compared with the healthy control group, the chronic hepatitis B, liver cirrhosis, and hepatocellular carcinoma groups had significant increases in the plasma levels of TGFβ1 (F=51.188, P＜0.001), HBx (F=39.227, P＜0.001), DBH (F=34431, P＜0.001), and HYP (F=16.211, P＜0.001), and a positive correlation was observed between plasma HBx and TGFβ1, between TGFβ1 and DBH, and between HYP and DBH (r=0.931, 0.863, and 0.765, all P＜0.001). ConclusionHBx protein can promote the secretion of TGFβ1 in LO2 cells, induce the proliferation and activation of LX-2 cells, promote the development of liver fibrosis, and upregulate the expression of TGFβ1 and DBH in LX-2 cells, and rhTGFβ1 stimulation can induce the activation of LX-2 cells and the upregulation of DBH expression.

全世界有超过2.4亿人感染乙型肝炎病毒(HBV),15％～40％的未接受治疗的慢性HBV感染者可进展为肝硬化,最终导致肝衰竭和肝癌[1-2].HBV是大小约为3.2 kb的部分环状双链DNA,其在DNA聚合酶作用下可转化形成共价闭合环状DNA (cccDNA).肝细胞中的两个高活性启动子EnhancerⅠ(En1)、 EnhancerⅡ(En2)与肝细胞表面表达的特异性受体蛋白及细胞内富集的各类转录调控因子共同决定了HBV的嗜肝特性[3].想通过药物彻底消灭慢性HBV携带者体内的全部病毒十分困难,因此,寻找新的HBV检测标志物长链非编码RNA(lncRNA)在早期诊断HBV感染及调控HBV的复制过程有着重要的意义.

BACKGROUND:This study aimed to develop a method for assessing the sensitivity and diagnostic performance of the neutrophil surface CD64 stimulation index (SI) in tuberculosis infection.METHODS:A total of 149 samples were divided into three groups (tuberculosis group, n = 51; nontuberculosis infection group, n = 50; and healthy control group, n = 48). Flow cytometry was used to detect the sensitivity of CD64 SI on the surface of neutrophils. The sensitivities of CD64 SI before and after stimulation with ESAT-6 and CFP-10 antigens were compared using interferon-gamma release assay-enzyme-linked immunosorbent assay (IGRA-ELISA).RESULTS:The diagnostic threshold for CD64 SI based on the receiver operating characteristic curve was found to be 2.025, which is the standard for judging tuberculosis infection. The IGRA-ELISA and the CD64 SI assays were highly consistent with a kappa value of 0.635 (p < 0.003, 95% CI: 0.002 - 0.003).CONCLUSIONS:The neutrophil surface CD64 SI value detection method may serve as one of the new diagnostic methods for active Mycobacterium tuberculosis infection.

Hepatitis B virus (HBV) infection remains a global public health problem. Nearly 257 million people worldwide have been infected with HBV, resulting in 887,000 people dying of cirrhosis or liver cancer caused by chronic hepatitis B (CHB) annually. Therefore, identification of new targets against HBV is urgently needed. Long noncoding RNAs (LncRNAs) have gained widespread attention in recent years due to their function in cancer, inflammation and other diseases. Notably, a growing number of lncRNAs have been found to play a role in HBV development. In the present study, we first identified a famous lncRNA, HOTAIR, which was significantly up-regulated in HBV-infected cells and PBMCs from CHB patients. Furthermore, we evaluated the clinical relevance of HOTAIR in 20 CHB patients and found that higher levels of HOTAIR expression were associated with higher ALT/AST levels and were positively correlated with HBsAg and HBV DNA levels. In addition, functional analysis showed that HOTAIR promoted HBV transcription and replication by elevating the activities of HBV promoters via modulation of the levels of cccDNA-bound SP1. In conclusion, our study reveals that HOTAIR expression is correlated with the clinicopathological and physiological characteristics of HBV. Thus, HOTAIR may serve as a novel HBV diagnostic and therapeutic biomarker based on its ability to facilitate HBV transcription and replication.

目的:分析卡氏肺孢子菌感染患者临床诊治及病原体的检测,探讨实验室卡氏肺孢子菌对六胺银染色法检测的影响因素及其镜下的特征.方法:抽取医院微生物室2017年9月-2018年3月间采用六胺银染色法检测的15例标本,分析其中1例急性淋巴细胞白血病患儿伴有肺孢子菌感染引起呼吸衰竭病例的诊治过程检测结果及肺孢子菌镜下的特征.结果:15例卡氏肺孢子菌六胺银染色标本中,其中阳性标本为6例;此6例病例均为免疫力低下的非艾滋病患者,由于检测病原体的及时和准确性,无死亡病例发生,其中2例转外院治疗,4例治愈后康复出院.结论:对于免疫力低下的肺炎患者,常规治疗无效的,应考虑可能是卡氏肺孢子菌感染.

目的 优化免疫组织化学法检测三阴乳腺癌(TNBC)组织中Ki-67抗原实验条件,实现检测技术标准化.方法 20例诊断为TNBC的组织标本,使用标准免疫组织化学检测Ki-67抗原.针对不理想的结果 对实验条件进行调整,直至获得最佳结果.使用图像分析软件对阳性染色细胞计数分析.结果通过对脱蜡 、孵一抗 、分化时间控制等因素的优化,得到更为理想的免疫组织化学结果.阳性细胞数和总细胞数通过软件灰度值分析来获得.最终20例标本均为Ki-67阳性高表达.结论 改良的免疫组织化学染色法对TNBC组织中Ki-67抗原检测结果较传统法更加精准 、结果更为可靠.

Background Acute meningitis and encephalitis syndromes (AMES) is a severe neurological infection which causes high case fatality and severe sequelae in children. To determine the etiology of childhood AMES in Shenzhen, a hospital-based study was undertaken. Methods A total of 240 cerebrospinal fluid (CSF) samples from 171 children meeting the case definition were included and screened for 12 common causative organisms. The clinical data and conventional testing results were collected and analyzed. Whole genome sequencing was performed on a Neisseria meningitidis isolate. Results A pathogen was found in 85 (49.7%) cases; Group B Streptococcus (GBS) was detected in 17 cases, Escherichia coli in 15, Streptococcus pneumoniae in 14, enterovirus (EV) in 13, herpes simplex virus (HSV) in 3, N. meningitidis in 1, Haemophilus influenzae in 1, and others in 23. Notably, HSV was found after 43 days of treatment. Twelve GBS and 6 E. coli meningitis were found in neonates aged less than 1 month; 13 pneumococcal meningitis in children aged > 3 months; and 12 EV infections in children aged > 1 year old. The multilocus sequence typing of serogroup B N. meningitidis isolate was ST-3200/CC4821. High resistance rate to tetracycline (75%), penicillin (75%), and trimethoprim/sulfamethoxazole (75%) was found in 4 of S. pneumoniae isolates; clindamycin (100%) and tetracycline (100%) in 9 of GBS; and ampicillin (75%) and trimethoprim/sulfamethoxazole (67%) in 12 of E. coli . Conclusions The prevalence of N. meningitidis and JEV was very low and the cases of childhood AMES were mainly caused by other pathogens. GBS and E. coli were the main causative organisms in neonates, while S. pneumoniae and EV were mainly found in older children. HSV could be persistently found in the CSF samples despite of the treatment. A better prevention strategy for GBS, the introduction of pneumococcal vaccine, and incorporation of PCR methods were recommended.

Gastric cancer (GC) is one of the leading causes of cancer-related death in both men and women due to delayed diagnosis and high metastatic frequency. Extracellular vesicles (EVs) are membrane-bound nanovesicles which are released by cells into body fluids such as plasma, saliva, breast milk, cerebrospinal fluid, semen, urine, lymphatic fluid, amniotic fluid, sputum and synovial fluid. EVs deliver almost all types of biomolecules such as proteins, nucleic acids, metabolites, and even pharmacological compounds. These bioactive molecules can be delivered to recipient cells to influence their biological properties, modify surrounding microenvironment and distant targets. The extensive exploration of EVs enhances our comprehension of GC biology referring to tumor growth, metastasis, immune response and evasion, chemoresistance and treatment. In this review, we will sum up the effects of GC-derived EVs to the tumor microenvironment. Moreover, we will also summarize the function of microenvironment-derived EVs in GC and discuss how the bidirectional communication between tumor and microenvironment affect GC growth, metastatic behavior, immune response, and drug resistance. At last, we prospect the clinical application viewpoint of EVs in GC.