Sepsis-induced acute respiratory distress syndrome (ARDS) is a life-threatening inflammatory lung condition with high mortality and no specific pharmacological treatments. The complex spatial heterogeneity of the lung during sepsis hinders the discovery of effective therapies. This study aimed to use spatial transcriptomics to map the septic lung’s molecular landscape to identify and validate a novel therapeutic agent. We performed spatial transcriptomics on lung tissues from mice subjected to cecal ligation and puncture (CLP) to model sepsis. A computational drug screen identified Mavacamten. In vitro, lipopolysaccharide-stimulated murine alveolar epithelial cells were used to assess Mavacamten’s effects on inflammation and cell injury. In vivo, CLP mice received Mavacamten, and we assessed survival, lung function, pulmonary edema, histology, and inflammatory markers. The underlying mechanism was investigated by analyzing the PI3K/AKT/mTOR pathway and autophagy markers. Statistical analyses included ANOVA, t-tests, and Kaplan-Meier analysis. Spatial transcriptomics revealed distinct cellular clusters that were dramatically rearranged during sepsis. In vitro, Mavacamten significantly attenuated lipopolysaccharide-induced inflammation, cellular injury, and oxidative stress. In the CLP mouse model, Mavacamten treatment markedly improved 7-day survival, restored arterial oxygenation, reduced pulmonary edema, and lessened histological lung injury. Mavacamten also significantly lowered local and systemic pro-inflammatory cytokine levels. Mechanistically, Mavacamten reversed the sepsis-induced inhibition of autophagy and suppressed the activation of the PI3K/AKT/mTOR signaling pathway in lung tissues. Mavacamten confers robust protection against sepsis-induced ARDS in a preclinical model by mitigating inflammation and lung injury, leading to improved survival. Its therapeutic action is mediated by inhibiting the PI3K/AKT pathway and restoring protective autophagy. Mavacamten is a promising candidate for repurposing in the treatment of sepsis-induced ARDS.

Aims:This study aimed to explore the genotype and phenotype correlation of patients with multiple osteochondroma (MO), and validate phenotypic differences in ATDC5 cell model with EXT1 or EXT2 gene disruption (EXT1-/- or EXT2-/-). Methods:Mutation analysis was employed in 27 families with MO using polymerase chain reaction (PCR)-Sanger sequencing and targeted next-generation sequencing (t-NGS). ATDC5 cell model with EXT1-/- or EXT2-/- was established using CRISPR/Cas9 technology. Markers of chondrocyte proliferation and differentiation were assessed through quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and dyeing experiments at different timepoints. Results:A total of 27 pathogenic mutations were identified in EXT1 and EXT2 genes, in which nine were novel mutations. In terms of mutation rate, onset age, medical treatment age, and numbers of moderate/severe (II/III) patients, the numbers with EXT1 gene mutations were more than those with EXT2 gene mutations. ATDC5 cell model with EXT1-/- or EXT2-/- was successfully established. The proliferation rate of ATDC5 chondrocytes in mutant groups (EXT1-/- and EXT2-/-) was higher than in WT group. The expressions of proliferation markers, including α-1 chain of type II collagen (COL2A1), aggrecan (ACAN), and SRY-box transcription factor 9 (SOX9), were almost all higher in mutant groups than that in WT group. Moreover, the expression of differentiation indicators, including α-1 chain of type X collagen (COL10A1), Runt-related transcription factor 2 (RUNX2), and matrix metallopeptidase 13 (MMP13), was increased in mutant groups compared to WT group. The Alcian blue and Alizarin red staining obtained similar results. Conclusion:Clinical research identified nine novel mutations in EXT1 and EXT2 genes from 27 families with MO, and patients with EXT1 mutations have more severe symptoms than with EXT2 mutations. Cellular experiments suggested that EXT1 and EXT2 gene mutations promote proliferation and differentiation of chondrocytes, which were consistent with clinical research.

Abstract: Background: Sepsis-Induced Myocardial Injury (SIMI) is a serious complication with high in-hospital and 28-day mortality. Dexmedetomidine (DEX), a selective α2-adrenergic receptor agonist, has shown anti-inflammatory and cardioprotective effects, but its impact on SIMI prognosis is unclear. Methods: This retrospective study used the MIMIC-IV database, including SIMI patients diagnosed per Sepsis-3 criteria. Propensity score matching (PSM) balanced baseline characteristics, and Cox regression analyzed the association between DEX use and mortality outcomes. External validation was performed using the MIMIC-III database. Results: Among 3,921 SIMI patients (375 DEX, 3,546 non-DEX), DEX use showed a trend toward lower 28-day mortality (HR 0.49, 95% CI 0.23-1.05, p = 0.068) and in-hospital mortality (HR 0.45, 95% CI 0.20-1.02, p = 0.055). After PSM, mortality remained lower in the DEX group. ICU and hospital stays were longer in the DEX group, but CRRT use showed no significant differences. External validation confirmed these findings. Conclusion: DEX use may reduce 28-day and in-hospital mortality in SIMI patients, providing preliminary evidence for its potential benefits. Further large-scale studies are needed to confirm these results. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement the National Traditional Chinese Medicine Priority Specialty Discipline Cultivation Program of China (2024-90). ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: https://mimic.mit.edu/docs/iv/ I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes The data used in this study were obtained from the publicly accessible MIMIC-IV database (https://mimic.mit.edu/docs/iv/), with all data anonymized to protect patient privacy.Research data are available in accordance with the database access policy.

Background: LncRNA taurine-upregulated gene 1 (TUG1) can regulate vascular endothelial cell injury, a critical mechanism in treating hemorrhagic shock and fluid resuscitation (HS/R). Therefore, this study explored the influence of TUG1 in HS/R. Methods: An in vivo rat model of ischemia-reperfusion (I/R) injury post-HS/R and an in vitro model of oxidative stress injury in rat cardiomyocyte cell line (H9C2) were constructed. In vivo, we silenced TUG1 and quantified its expression along with inflammatory factors through quantitative reverse transcription polymerase chain reaction (qRT-PCR), mean arterial pressure (MAP) detection and blood gas analysis. Myocardial functional impairment was assessed via Triphenyl-2H-Tetrazolium Chloride (TTC), Hematoxylin and eosin, and Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) stainings. Oxidative stress level in rat serum was measured. In vitro, we examined the changes of cell viability, apoptosis, oxidative stress levels, inflammatory factor secretion and nuclear factor-κB (NF-κB)/p65 expression by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), flow cytometry, Enzyme-linked immunosorbent assay (ELISA) and Western blot. Results: TUG1 level was elevated in rats of I/R model caused by HS/R. TUG1 silencing ameliorated the decline in MAP, acid-base imbalance and myocardial tissue damage, and suppressed oxidative stress and inflammatory factor levels in model rat. TUG1 silencing enhanced viability, impeded apoptosis, and reduced oxidative stress, inflammatory factor contents and NF-κB/p65 expression in H2O2 treated H9C2 cells. Conclusion: TUG1 participates in regulating oxidative stress damage and inflammation induced by HS/R.

Background:Xijiao Dihuang decoction (XJDHT), a traditional Chinese medicine, is widely used to treat patients with sepsis. However, the mechanisms underlying the effects of XJDHT on cardiac dysfunction have yet to be fully elucidated. The present study evaluated the potential utility of XJDHT in protecting against sepsis-induced cardiac dysfunction and myocardial injury.Methods:The mice were randomly divided into 3 groups and administered Lipopolysaccharide (LPS,10 mg/kg) or equivalent saline solution (control) and treated with XJDHT (10 g/kg/day) or saline by gavage for 72 hours. XJDHT was dissolved in 0.9% sodium chloride and administered at 200 μL per mouse. Transthoracic echocardiography, RNA-seq, TUNEL assays and hematoxylin and eosin (H&E) staining of cardiac tissues were performed.Results:Treatment with XJDHT significantly enhanced myocardial function and attenuated pathological change, infiltration of inflammatory cells, levels of TNF-α, IL-1β and expression of TLR4 and NF-κB in mice with sepsis. RNA sequencing and Kyoto Encyclopedia of Genes and Genomes pathway analyses identified 531 differentially expressed genes and multiple enriched signaling pathways including the PI3K/AKT pathway. Further, XJDHT attenuated cardiac apoptosis and decreased Bax protein expression while increasing protein levels of Bcl-2, PI3K, and p-AKT in cardiac tissues of mice with sepsis.Conclusion:In summary, XJDHT improves cardiac function in a murine model of sepsis by attenuating cardiac inflammation and apoptosis via suppressing the TLR4/NF-κB pathway and activating the PI3K/AKT pathway.

Background This study aims to develop and validate a novel mortality prediction model to forecast the 90-day mortality risk for patients with ACS (Acute Coronary Syndrome) after discharge. Methods We selected 1359 patients from the Medical Information Mart for Intensive Care IV (MIMIC-IV) database as our study cohort and collected 32 clinical indicators within the first 24 hours of their admission. By randomly assigning these patients to a training group and a validation group (with a ratio of 0.65:0.35), we used Least Absolute Shrinkage and Selection Operator (LASSO) regression and bidirectional stepwise logistic regression to identify 7 key variables. Based on these variables, we constructed a mortality prediction model. To evaluate the model's accuracy and reliability, we plotted the Receiver Operating Characteristic (ROC) curve, calculated the Area Under the Curve (AUC), sensitivity, and specificity, and performed calibration analysis, including plotting calibration curves, calculating Brier scores, and conducting Hosmer-Lemeshow goodness-of-fit tests. Additionally, through Decision Curve Analysis (DCA) and comparison with current clinical scoring systems, we further assessed the clinical utility of our model. Results Age, SOFA (Sepsis-related Organ Failure Assessment), APS III (Acute Physiology Score III), AG(Anion Gap), RR(Respiratory rate), INR(International normalized ratio), and BUN(Bun urea nitrogen) were identified as independent predictors of 90-day mortality risk. The model demonstrated good diagnostic performance in both the training and validation groups, with AUC values of 0.842 and 0.855, respectively. The Hosmer-Lemeshow test results indicated a good fit for both datasets, with P-values of 0.1626 and 0.4008. The Brier scores were 0.107 for the training set and 0.103 for the validation set, indicating the model's good predictive performance. Compared to existing scoring systems (SOFA, APSIII), DCA showed that our model could provide a higher net benefit in clinical applications. Conclusion We identified seven clinical indicators including age, SOFA, APSIII, AG, RR, INR, and BUN as independent prognostic factors for predicting the 90-day all-cause mortality in patients with ACS after discharge. This model can assist ICU physicians to quickly make preliminary clinical decisions for ACS patients in clinical practice.

目的 通过CYP2C19基因多态性的检测数据,筛选使用过伏立康唑的患者,探讨CYP2C19基因多态性与伏立康唑疗效之间是否具有临床关系,用以指导伏立康唑在临床上的个体化用药.方法 回顾性分析我院检测过CYP2C19基因型且使用伏立康唑的患者84例,通过设计疗效与不良反应量表对其进行评分,分析伏立康唑的疗效与CYP2C19基因型之间的关系.结果 伏立康唑的疗效在快代谢与中代谢的患者中较佳,慢代谢者疗效较差,差异有统计学意义(P＜0.05);不良反应比较,差异无统计学意义(P＞0.05).结论 伏立康唑的疗效与患者CYP2C19的基因型具有一定的临床关系,慢代谢患者使用伏立康唑的抗感染效果低于快、中代谢者,CYP2C19基因多态性对伏立康唑临床应用的影响主要体现在对伏立康唑给药方案的调整上,尤其是给药剂量的调整.

Objective: Polydactyly is characterized by multiple distinct heterogeneous phenotypes, the etiologies of which involve several genes. This study aimed to explore the genetic defects and further clarify the molecular mechanism of polydactyly in several Chinese families.Methods: Three families with diverse phenotypes of non-syndromic polydactyly were analyzed: two were cases of familial disease, whereas one was sporadic. PCR and Sanger sequencing were used to screen for pathogenic mutations in two known disease-associated genes, GLI3 and HOXD13, while bioinformatic analyses predicted the pathogenicity of the identified variants. Reverse transcription PCR was used to analyze the splicing effect of an intronic variant.Results: Two novel heterozygous frameshift mutations (c.4478delG/p.S1493Tfs*18; c.846_c.847insC/p.R283Qfs*21) were identified in the GLI3 gene from two of the pedigrees. Both c.4478delG and c.846_c.847insC were later confirmed in affected and unaffected members and normal controls, to truncate and disrupt the integrity of the GLI3 protein, reduce its level of expression, and disrupt its biological function through nonsense-mediated mRNA decay (NMD). In addition, a deep intron mutation (c.125-47 C>A) was detected in the GLI3 gene from the sporadic case, however, both bioinformatics analysis (HSF, splice AI, and CBS) and RT-PCR indicated that the variant c.125-47 C>A had minimal if any impact on splicing of the GLI3 gene.Conclusion: Two newly identified heterozygous frameshift mutations in the GLI3 gene were detected in two families with non-syndromic polydactyly, further extending the mutational spectrum of the GLI3 gene in non-syndromic polydactyly. Moreover, our study further expanded the phenotypic spectrum of non-syndromic polydactyly.

目的 采用小RNA(sRNA)测序技术检测清瘟败毒饮干预的脓毒症小鼠24?h脾组织微小RNA(miRNA)的表达变化,从而从非编码基因层面分析清瘟败毒饮抗脓毒性脾功能损伤的可能机制.方法 将45只SPF级健康BALB/C小鼠随机等分为对照组、脓毒症组和清瘟败毒饮组.清瘟败毒饮成分每剂含生石膏15?g、生地黄7.5?g、水牛角15?g、黄连2.5?g、栀子5?g、桔梗5?g、黄芩5?g、知母5?g、赤芍5?g、玄参5?g、连翘5?g、甘草5?g、丹皮5?g、竹叶5?g.脓毒症组和清瘟败毒饮组小鼠腹腔注射脂多糖(LPS)10?mg/kg构建脓毒症模型,对照组腹腔注射等量生理盐水.清瘟败毒饮组于制模前2?d开始给予清瘟败毒饮药液0.3?mL,每1?mL相当于生药1.5?g灌胃,术后继续胃饲,每日2次;对照组及脓毒症组给予等量生理盐水胃饲.术后24?h各组随机取5只小鼠脾组织,每组各取1只小鼠的脾组织于光镜下观察病理学改变,提取其余小鼠脾组织总RNA,采用sRNA测序技术进行miRNA检测,应用生物信息学软件分析脓毒症小鼠脾组织miRNA表达差异及靶基因的预测.?结果 光镜下显示,对照组小鼠脾组织结构正常;脓毒症组小鼠脾组织白髓总体结构稍紊乱,红白髓交界偏模糊,红髓充血明显;清瘟败毒饮组小鼠脾组织紊乱结构较前有所改善.与对照组比较,脓毒症组小鼠脾组织已知miRNA表达上调数为39个,下调数为16个;清瘟败毒饮组小鼠脾组织已知miRNA表达上调数为6个,下调数为24个.经LPS干预后,miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p表达明显上调;经清瘟败毒饮干预后,miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p表达出现一定程度下调.结论 清瘟败毒饮可改善脓毒症小鼠脾组织结构改变,可能通过下调miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p基因表达起到保护脓毒症小鼠脾损伤的作用,可为将来进一步研究提供思路.

OBJECTIVE To investigate the effect of CYP2C9 gene polymorphism on drug anticoagulant therapy and gastrointestinal adverse reactions in patients with coronary heart disease complicated with atrial fibrillation in a hospital, provide clinical evidence for rational drug use.METHODS From January 2018 to January 2020,33 patients in the top three hospitals who were diagnosed with coronary heart disease complicated with atrial fibrillation were enrolled in the study.Detect its genotype, the effect of drug anticoagulant treatment, the combination of drug use and the occurrence of gastrointestinal symptoms were observed.RESULTS The proportions of complete remission, partial remission and poor curative effect in patients with different CYP2C9 gene metabolism types were significantly different（P=0.025）,There was also a statistically significant difference in the composition ratio of curative effects among different drug groups（P=0.004）.There were statistically significant difference in the composition ratio of patients with gastrointestinal symptoms in different metabolic types and different combination regimen groups.CONCLUSION The type of CYP2C9 gene mutation has an impact on the effect of drug anticoagulant therapy and the appearance of gastrointestinal symptoms in patients with coronary heart disease complicated with atrial fibrillation.

Objective:To explore the mechanism of Xijiao Dihuang Ddecoction (XJDHT) against sepsis-induced liver injury based on transcriptomics.Methods:Sixty C57BL/6 mice were randomly (random number) divided into the sepsis group, sepsis treatment with XJDHT and control group, with 20 mice in each group. The sepsis mouse model was established by intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). The control group was intraperitoneally injected with the same amount of normal saline. The sepsis treatment with XJDHT group was injected with XJDHT (crude drug 187.5 mg) twice a day 2 days before modeling. After modeling, gastric feeding was continued twice a day, while the control group and sepsis group were gavaged with the same amount of normal saline. At 72 h after LPS intervention, 9 mice in each group were randomly selected. After anesthesia, part of the liver were taken for small RNA and RNA sequencing and analysis, and part of the liver were taken for pathological examination.Results:XJDHT could improve the histopathological changes of liver in septic mice, and alleviate some abnormally expressed microRNAs (mmu-mir-292a-5p, mmu-mir-871-3p, mmu-mir-653-5p, mmu-mir-293-5p, mmu-mir-155-3p, mmu-mir-346-5p, mmu-mir-187-5p, mmu-mir-3090-3p) and their target genes.Conclusions:XJDHT can reduce the liver histopathological changes in septic mice, and its mechanism may be related to XJDHT regulating the expression of important key genes of liver of sepsis like mmu-mir-187-5p and its target genes such as ADAM8, irak3 and PFKFB3

目的 制备穿心莲内酯MPEG-PDLLA(聚乙二醇单甲醚聚丙交酯)胶束使穿心莲内酯被包裹而溶解于水并对其进行含量测定.方法 采用薄膜蒸发法制备穿心莲内酯MPEG-PDLLA胶束;紫外分光光度计法测定含量、包封率;采用马尔文ZS90粒度仪测定胶束的粒径.结果 制备的穿心莲内酯MPEG-PDLLA胶束水溶性好、水溶液澄清透明,平均粒径约为27 nm.结论 难溶性药物穿心莲内酯制成可溶于水的胶束,有望成为穿心莲内酯的新剂型.

目的 采用Small RNA测序技术检测脓毒症小鼠24 h脾组织microRNA(miRNA)的表达变化,并从非编码基因层面分析脓毒症脾功能损伤的可能机制.方法 将30只SPF级健康BALB/C小鼠随机分为对照组和脓毒症组,每组15只.脓毒症组采用LPS 10 mg/kg腹腔注射进行建模,对照组仅予等量生理盐水腹腔注射.两组术毕均行肌肉注射平衡液5 ml/kg,并于术后24 h各组随机取5只小鼠的脾组织提取RNA,采用Small RNA测序技术进行miRNA检测,然后应用生物信息学软件分析脓毒症小鼠脾组织的miRNA表达差异及靶基因预测.结果 小鼠建模后24 h,与对照组相比,脓毒症小鼠脾脏组织已知及未知miRNA表达上调数分别为39个、3个,下调数分别为16个、3个.其中已知miRNA表达上、下调倍数最大的前5个基因分别为Rno-miR-217-5p,rno-mir-216a-5p,rnomiR-216b-5p,rno-miR-375-3p,rno-mir-216b-3p.下调倍数前5的miRNA:Rno-miR-138-5p,rno-miR-451a-3p,rno-miR-138-1-3p,rno-miR-202-5p,rno-miR-592.结论 LPS诱导脓毒症小鼠可出现miRNA表达量的改变,其上、下调倍数最显著的miRNA可能对预测LPS诱导的脓毒症脾功能损伤有一定指导意义.

OBJECTIVE:To explore the genetic basis for a pedigree affected with hereditary multiple osteochondroma (HMO).METHODS:Peripheral blood samples were collected from the proband and members of his pedigree with informed consent. Following extraction of genomic DNA, all coding exons and flanking intronic sequences (-10 bp) of the EXT1 and EXT2 genes were subjected to targeted capture and next generation sequencing (NGS). Suspected variant was verified by Sanger sequencing.RESULTS:A heterozygous nonsense variant (c.1911C>A) was found in exon 10 of the EXT1 gene in the proband and his affected father but not in a healthy sister and normal controls. The variant was classified as a pathogenic based on the guidelines of the American College of Medical Genetics and Genomics (PVS1+PM2+PP1). Bioinformatic analysis predicted that the c.1911C>A variant may be disease-causing via nonsense-mediated mRNA decay and anomalous splicing.CONCLUSION:The c.1911C>A variant probably underlay the disease in this pedigree. Discovery of this variant enriched the variant spectrum of HMO.

目的 对比脓毒症组小鼠与正常对照组小鼠的肠道菌群,以明确肠道菌群组成特点.方法 将60只BALB/C小鼠随机等分为对照组(CF组)和脓毒症组(SF组)(每组各30只).SF组小鼠按脂多糖(LPS)10 mg/kg腹腔注射构建脓毒症模型;CF组则予等量生理盐水腹腔注射.术后24、72 h每组小鼠留取粪便标本行16sRNA测序.在72 h同时取盲肠组织做组织学检查.结果 LPS干预后72 h小鼠结肠局部黏膜上皮缺失,肠腺结构消失,炎性细胞浸润,上皮细胞肿胀.肠道菌群方面:SF组较CF组小鼠的肠道菌群多样性明显减少.CF组小鼠肠道菌群主要以拟杆菌门和厚壁菌门为主,但SF组小鼠厚壁菌属比例总体呈现减少趋势,变形菌门则表现为增长的趋势.LPS干预后24 h SF组小鼠肠道菌群出现紊乱:粪肠球菌属、大肠杆菌属、弗格森埃希菌属、缓慢葡萄球菌属及不可培养的大肠杆菌属明显增多(P<0.05),梭状芽孢菌属、毛螺科菌属、未培养的梭菌属则明显减少(P<0.05).LPS干预后72 h SF组小鼠肠道菌群的变化情况:铅黄肠球菌、粪肠球菌、大肠杆菌、费格森埃希菌、金氏拟杆菌、奇异变形杆菌、未培养的埃希菌属和肠球菌明显增多(P<0.05),鼠乳杆菌、未培养的拟杆菌属、约氏乳杆菌则明显减少(P<0.05).结论 脓毒症小鼠可致肠道微生态的紊乱,微生物的多样性减少,潜在致病菌的过度生长,同时炎症作用下肠道屏障的破坏可能为微生物移位创造条件,也可能成为病情进一步发展的重要推动力.

目的 基于mRNA测序(RNA-seq)技术,采用生物学信息分析软件分析清瘟败毒饮对脓毒症大鼠心肌组织中白细胞介素-17(IL-17)信号转导通路相关基因表达的变化,阐明清瘟败毒饮对脓毒症心肌的保护机制.方法 清洁级健康雄性Wistar大鼠45只,随机分为脓毒症模型组、清瘟败毒饮组和假手术组,每组15只.采用盲肠结扎穿孔术(CLP)制作脓毒症大鼠模型,假手术组仅开腹、关腹与复苏,不进行CLP.清瘟败毒饮组大鼠在制模前2?d给予清瘟败毒饮(组成:生地黄7.5?g,生石膏、水牛角各15?g,黄连2.5?g,栀子、桔梗、黄芩、知母、赤芍、玄参、连翘、甘草、丹皮、竹叶各5?g)胃饲,每日2次,术后继续中药灌胃24?h.3组术毕均予5?mL/kg平衡液肌肉注射.术后24?h取大鼠肌组织提取总RNA,采用RNA-seq技术对mRNA表达量进行检测,并应用京都基因和基因组数据库通路富集方法(KEGG?Pathway)分析清瘟败毒饮对脓毒症大鼠心肌组织IL-17信号通路相关基因表达的影响.结果 RNA-seq检测结果显示,与假手术组相比,脓毒症模型组大鼠心肌组织表达上调的基因有900个,下调的基因有1507个;清瘟败毒饮组大鼠心肌组织表达上调的基因有1288个,下调的基因有1870个.KEGG?Pathway分析结果显示,与假手术组相比,脓毒症模型组大鼠心肌组织IL-17信号通路相关基因表达上调的有10个,下调的有5个;清瘟败毒饮组大鼠心肌组织IL-17信号通路基因表达上调的有11个,下调的有6个.其中脓毒症模型组及清瘟败毒饮组同时上调的基因有7个〔CXC趋化因子配体(CXCL1、CXCL2)、白细胞介素-6(IL-6)、环氧合酶-2(COX-2)、粒细胞集落刺激因子(G-CSF)、基质金属蛋白酶9?(MMP-9)、CCAAT/增强子结合蛋白β(C/EBPβ)〕;同时下调的基因有5个〔转化生长因子激酶1(TAK1)、核转录因子-κB(NF-κB)、泛素特异性蛋白酶25(USP25)、细胞外信号调节激酶(ERK)、天冬氨酸特异性半胱氨酸蛋白酶(CASP)〕;脓毒症模型组上调而清瘟败毒饮组正常表达的基因有2个〔CXCL5、白细胞介素-1β(IL-1β)〕;?脓毒症模型组下调而清瘟败毒饮组正常表达的基因有1个〔热休克蛋白90(HSP90)〕;脓毒症模型组正常表达而清瘟败毒饮组上调的基因有3个〔激活蛋白-1(AP-1)、IκB激酶-β(IKK-β)、CC趋化因子7(CCL7)〕;脓毒症模型组正常表达而清瘟败毒饮组下调的基因有1个(ERK);脓毒症模型组上调而清瘟败毒饮组下调的基因有1个〔IκB激酶-α(IKK-α)〕;脓毒症模型组下调而清瘟败毒饮组上调的基因有1个〔NF-κB抑制因子-α(IκB-α)〕.结论 清瘟败毒饮可通过调节IL-17信号通路相关基因的表达,对脓毒症大鼠心脏起保护作用.

目的 采用16sRNA测序技术检测大黄对脓毒症小鼠早期肠道菌群的变化情况,分析大黄对脓毒症小鼠的治疗作用机制.方法 将45只BALB/C小鼠随机等分为3组,即对照组、脓毒症组、脓毒症大黄干预组.实验前正常饮食、饮水.术前2 d脓毒症大黄干预组按生药量50 mg/kg灌胃给药,每天2次;对照组、脓毒症组给予等量生理盐水灌胃.3组小鼠灌胃给药2 d后,脓毒症组及脓毒症大黄干预组小鼠按LPS 10 mg/kg腹腔注射构建脓毒症模型,对照组予等量生理盐水腹腔注射.建模后24 h取盲肠组织行病理检查以及粪便组织16sRNA检测.结果 经大黄干预后,脓毒症小鼠盲肠组织结构的改变得到一定程度的缓解.脓毒症组小鼠厚壁菌属比例明显减少,拟杆菌属相对增多.脓毒症大黄干预组小鼠拟杆菌属、厚壁菌属、弯曲菌属均有减少,而变形菌属明显增多.结论 大黄干预后,脓毒症小鼠肠道菌群进一步变化,变形杆菌属相对增多为优势菌群;结合病理结果,考虑大黄对脓毒症小鼠的治疗作用主要是通过保护肠道黏膜屏障而起到保护作用.

Background: Multiple osteochondroma (MO), an autosomal dominant genetic disease, is caused by heterozygous mutations in the EXT1 and EXT2 genes. Approximately 80% of pathogenic mutations are nonsense/missense mutations, small indels, and splicing mutations. Splicing mutations, particularly at the 3 ' and 5 ' splice sites, disrupt normal mRNA processing and cause exon skipping or aberrant splicing, ultimately resulting in protein truncation and loss of function. Methods: Polymerase chain reaction (PCR) and Sanger sequencing were applied to detect subtle mutations in a Chinese family with MO, the pathogenicity of a splicing variant was predicted by bioinformatics and further verified using a minigene splicing assay. Results: A novel and heterozygous splicing mutation, c.626 + 2_626 + 5delTAGG, was identified in the EXT2 gene of the proband and the father by PCR and Sanger sequencing, whereas the unaffected mother and brother had wild-type alleles at the same site. Bioinformatics predicted that the 5 ' splicing site of exon 3 in the EXT2 gene was destroyed due to this mutation. A hybrid minigene splicing assay (HMSA) indicated that the mutation disturbed the normal splicing of the EXT2 gene mRNA and led to a deletion of 79 bp at the 5 ' end of exon 3, which resulted in aberrant splicing of exon 3 and introduced an earlier stop codon in the EXT2 gene. Conclusion: A novel splicing mutation was identified that produced the MO phenotype through aberrant splicing in a Chinese family. This observation, expands our knowledge of the spectrum of molecular pathogenic mechanisms leading to aberrant mRNA splicing.

目的 探讨犀角地黄汤对脓毒症大鼠脾脏组织NF-κB信号通路相关基因表达的影响.方法 将45只健康雄性Wistar大鼠随机为为对照组、脓毒症组、犀角地黄汤干预组,各15只.脓毒症组采用盲肠结扎穿孔术(CLP)建模.犀角地黄汤干预组大鼠除了CLP外,术前2天开始中药胃饲,每天上、下午各给药1次,术后连续胃饲2d.对照组仅行开腹、关腹,但不予盲肠结扎穿孔.3组术毕均肌肉注射平衡液5 mL/kg.术后48 h取脾组织提取RNA后采用RNA-seq进行基因检测,应用生物信息学方法分析脓毒症大鼠犀角地黄汤干预后NF-κB信号通路相关基因的表达变化.结果 脓毒症组及犀角地黄汤干预组,脾组织NF-κB信号通路相关基因出现同时表达上调的有2个,同时下调有18个,脓毒症组上调而犀角地黄汤干预组正常表达的有3个,仅脓毒症组下调而犀角地黄汤干预组正常表达的有5个,脓毒症组正常表达而犀角地黄汤干预组表达上调有4个(IL-1β、TNFα、MIP-1β、ICAM).结论 犀角地黄汤可能通过调节脓毒症大鼠脾组织NF-κB信号通路相关基因IL-1β、TNFα、MIP-1β、ICAM表达起到改善脓毒症晚期脾功能障碍的作用.

目的 基于TNF信号通路探讨清瘟败毒饮抗脓毒性脾功能损伤的可能机制.方法 选择清洁级健康雄性Wistar大鼠45只,随机分为对照组、脓毒症组(A组)、清瘟败毒饮干预组(B组),每组15只.A组大鼠采用盲肠结扎穿孔术(CLP)进行复制脓毒症模型;B组大鼠除CLP外,于术前2天给予中药胃饲2次/d,术后继续中药灌胃;对照组实施除CLP外的开腹、关腹.3组术毕均予5 ml/kg平衡液肌肉注射.术后24 h取脾组织提取总RNA后行RNA-seq对mRNA表达量进行检测,并应用生物信息学方法分析清瘟败毒饮对脓毒症大鼠脾组织TNF信号通路相关基因的表达变化.结果 相对于对照组,A、B组大鼠脾组织TNF信号通路相关基因中,同时上调的基因有19个,同时下调的基因有8个,下调转正常的基因1个,正常转上调的基因5个,由上调转为正常的基因7个(Ccl20、Cxcl10、IL-1b、IL6、Bcl3、Mmp9、Sele),正常转为下调的基因13个(ASK1、IKKα、CASP8、ITCH、p38、Drp1、IKKs、p38、Ccl5、Csf1、IL18R1、IL15、Ifi47).结论 清瘟败毒饮可通过调节TNF信号通路相关基因的过度表达,从而对脓毒症大鼠脾功能起到保护作用.