Ethnopharmacological relevance: Reynoutria japonica Houtt is a medicinal plant renowned for its diverse pharmacological properties, including heat-clearing, toxin-removing, blood circulation promotion, blood stasis removal, diuretic action, and pain relief. The plant is commonly utilized in Traditional Chinese Medicine (TCM), and its major bioactive constituents consist of polydatin (PD) and resveratrol (RES). Aim of the study: To summarize the relevant targets of PD in various oxidative stress-related diseases through the activation of Silence information regulator1 (SIRT1). Furthermore, elucidating the pharmacological effects and signaling mechanisms to establish the basis for PD's secure clinical implementation and expanded range of application. Materials and methods: Literature published before November 2023 on the structural analysis and pharmacological activities of PD was collected using online databases such as Google Scholar, PubMed, and Web of Science. The keywords were "polydatin", "SIRT1" and "oxidative stress". The inclusion criteria were research articles published in English, including in vivo and in vitro experiments and clinical studies. Non-research articles such as reviews, meta-analyses, and letters were excluded. Results: PD has been found to have significantly protective and curative effects on diseases associated with oxidative stress by regulating SIRT1-related targets including peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1 alpha), nuclear factor erythroid2-related factor 2 (Nrf2), high mobility group box 1 protein (HMGB1), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), p38/p53, as well as endothelial nitric oxide synthase (eNOs), among others. Strong evidence suggests that PD is an effective natural product for treating diseases related to oxidative stress. Conclusion: PD holds promise as an effective treatment for a wide range of diseases, with SIRT1-mediated oxidative stress as its potential pathway.

Based on network pharmacology and in vitro experimental verification, this study was designed to explore the potential mechanism of polydatin（PD） against bacterial pneumonia from the point of its polarization modulation on macrophages. Drug targets were predicted by SwissTargetPrediction, SEA, TCMSP and PharmMapper databases; disease targets were obtained in GeneCards and DisGeNET databases; GO and KEGG enrichment analysis were established by DAVID databases; protein interaction networks were constructed using STRING databases. And in vitro cell experiments were carried out to further verify the results of network pharmacology. The RAW264.7 cells were divided into three groups: model control group（model, M） stimulated with LPS, PD experimental group treated with PD, and blank control group（Control, C）. CCK8 was used to detect the effect of different concentrations of PD on cell viability; ELISA was used to detect the secretion levels of NF-κB and IL-1β, the terminal effector molecules of SIRT1/HMGB1 signaling pathway, and the macrophage polarization factors MCP-1 and Arg1 in the cell supernatant; qPCR was used to detect the mRNA expression of SIRT1/HMGB1 signaling pathway related molecules SIRT1, HMGB1, TLR4 and TRAF6 in cell pellets. According to the results of network pharmacological analysis, there are 61 intersection targets were screened out, in which SIRT1, HMGB1, TLR4, TRAF6 and NF-κB were identified as subsequent experimental targets by combining the results of GO and KEGG enrichment analysis and protein interaction network construction.The results of cell viability experiments confirmed that the secretion levels of NF-κB, IL-1β and MCP-1 in the PD group were significantly lower, while Arg1 was significantly higher than those in the M group; the mRNA expression of SIRT1 in PD group was significantly up-regulated, but the mRNA expressions of HMGB1, TLR4 and TRAF6were significantly down-regulated. In summary, polydatin can regulate the expression of signaling pathway molecules such as TLR4 and NF-κB and polarizing factors MCP-1 and Arg1 through SIRT1/HMGB1, and modulate macrophage polarization, thus may be a potential drug for the treatment of bacterial pneumonia.

MicroRNAs (miRNAs), which were initially discovered in Caenorhabditis elegans, can regulate gene expression by recognizing cognate sequences and interfering with the transcriptional or translational machinery. The application of bioinformatics tools for structural analysis and target prediction has largely driven the investigation of certain miRNAs. Notably, it has been found that certain miRNAs which are widely involved in the inflammatory response and immune regulation are closely associated with the occurrence, development, and outcome of bacterial pneumonia. It has been shown that certain miRNA techniques can be used to identify related targets and explore associated signal transduction pathways. This enhances the understanding of bacterial pneumonia, notably for "refractory" or drug-resistant bacterial pneumonia. Although these miRNA-based methods may provide a basis for the clinical diagnosis and treatment of this disease, they still face various challenges, such as low sensitivity, poor specificity, low silencing efficiency, off-target effects, and toxic reactions. The opportunities and challenges of these methods have been completely reviewed, notably in bacterial pneumonia. With the continuous improvement of the current technology, the miRNA-based methods may surmount the aforementioned limitations, providing promising support for the clinical diagnosis and treatment of "refractory" or drug-resistant bacterial pneumonia.

As far the current severe coronavirus disease 2019 (COVID-19), respiratory disease is still the biggest threat to human health. In addition, infectious respiratory diseases are particularly prominent. In addition to killing and clearing the infection pathogen directly, regulating the immune responses against the pathogens is also an important therapeutic modality. Sirtuins belong to NAD+-dependent class III histone deacetylases. Among 7 types of sirtuins, silent information regulator type-1 (SIRT1) played a multitasking role in modulating a wide range of physiological processes, including oxidative stress, inflammation, cell apoptosis, autophagy, antibacterial and antiviral functions. It showed a critical effect in regulating immune responses by deacetylation modification, especially through high-mobility group box 1 (HMGB1), a core molecule regulating the immune system. SIRT1 was associated with many respiratory diseases, including COVID-19 infection, bacterial pneumonia, tuberculosis, and so on. Here, we reviewed the latest research progress regarding the effects of SIRT1 on immune system in respiratory diseases. First, the structure and catalytic characteristics of SIRT1 were introduced. Next, the roles of SIRT1, and the mechanisms underlying the immune regulatory effect through HMGB1, as well as the specific activators/inhibitors of SIRT1, were elaborated. Finally, the multitasking roles of SIRT1 in several respiratory diseases were discussed separately. Taken together, this review implied that SIRT1 could serve as a promising specific therapeutic target for the treatment of respiratory diseases.

目的 初步探索芪归银方(Qiguiyin,QGY)对脂多糖(lipopolysaccharide,LPS)诱导的大鼠肺泡巨噬细胞NR8383免疫功能的调节作用,为揭示其抗炎分子机制提供实验基础.方法 以LPS刺激大鼠肺泡巨噬细胞NR8383,构建体外炎性细胞模型;设置空白对照组(control组)、模型对照组(LPS组)和芪归银实验组(LPS+QGY组),以CCK-8法检测QGY干预24、48及72h时巨噬细胞活力;以ELISA法检测QGY干预24、48及72h时NR8383炎性细胞上清液中白介素(interleukin,IL)-1β、肿瘤坏死因子(tumor necrosis factor,TNF)-α、单核细胞趋化蛋白(monocyte chemoattractant protein,MCP)-1和大鼠精氨酸酶(arginase,Arg)-1等免疫因子的浓度.结果 以1μg/ml的LPS成功构建了NR8383炎性细胞模型.0.125～2.000mg/ml的QGY在72h内对NR8383巨噬细胞无明显毒性不良反应;1～2mg/ml的QGY显著促进NR8383巨噬细胞的增殖(P<0.05).与control组比较,LPS组中IL-1β、TNF-α和MCP-1的含量均显著增高(P均<0.05),Arg-1的含量差异无统计学意义(P>0.05).与control组比较,1～2mg/ml的QGY可显著降低IL-1β和MCP-1的含量(P均<0.05),2mg/ml则可显著降低TNF-α 的含量(P<0.05).其中,2mg/ml的QGY作用48h,可显著降低TNF-α 和MCP-1的含量(P均<0.01).与control组比较,QGY对NR8383巨噬细胞Arg-1的含量比较,差异无统计学意义(P>0.05).结论 QGY对大鼠肺泡巨噬细胞NR8383有明确的免疫调节作用.

白藜芦醇对革兰氏阴性及阳性菌、病毒、真菌及衣原体等抑制作用明确,可通过干扰细菌生物膜的形成、抑制药物外排泵的功能、增加细胞的通透性及修饰药物作用靶点等方面参与细菌耐药的调控.此外,白藜芦醇还可作为抗菌增效剂增加细菌对抗生素的敏感性,提高抗生素的疗效.白藜芦醇治疗耐药菌感染有多重优势,如不易引起细菌耐药、成分较为明确、易获取、抑菌谱广泛、抑菌机制多样、可协同增效抗生素等.然而,结合中药免疫调理特点阐释白藜芦醇对细菌耐药机制改变的报道尚不多见,可进一步加强结合HMGB1这一免疫网络核心调控分子的研究.此外,白藜芦醇及虎杖苷胃肠道吸收良好,为其给药方式及临床口服剂型的探索提供了保障,提示基于病原体/机体免疫,全面揭示了白藜芦醇调控细菌耐药机制的单体中药研发的迫切必要及潜在可能.

Ferroptosis is a recently identified iron-dependent form of nonapoptotic cell death characterized by reactive oxygen species (ROS) generation and lipid peroxidation. Here, we report a novel iron-dependent form of ferroptosis induced by labile iron and investigate the mechanism underlying this process. We find that labile iron-induced ferroptosis is distinct from canonical ferroptosis and is linked to the mitochondrial pathway. Specifically, the mitochondrial calcium uniporter mediates the ferroptosis induced by labile iron. Interestingly, cells undergoing labile iron-induced ferroptosis exhibit cytoplasmic features of oncosis and nuclear features of apoptosis. Furthermore, labile iron-induced ferroptosis involves a unique set of genes. Finally, labile iron-induced ferroptosis was observed in liver subjected to acute iron overload in vivo. Our study reveals a novel form of ferroptosis that may be implicated in diseases caused by acute injury.

Aim: This study aimed to determine the effect of mechanical ventilation (MV) on the differentiation and proliferation of diaphragm satellite cells. Methods: Diaphragm satellite cells were isolated from C57 mice receiving 6 h of MV with optimized magnetic-activated cell sorting (MACS) approach. The cells were stained with BrdU or antibody for differentiation marker MYH3. The expression of MyoD and myogenin was detected by real-time PCR. Results: Diaphragm satellite cells were successfully isolated from mice by using MACS with a set of optimized parameters. About 1.5 × 105 cells could be harvested from a diaphragm. Upon MV, the proliferation rate of diaphragm satellite cells was decreased from 88.74% to 81.92%, while the differentiation rate was increased from 17.94% to 27.58%, compared to controls. Moreover, the expression of MyoD and myogenin were significantly upregulated upon MV. Conclusions: We established a practical method to purify diaphragm satellite cells, and demonstrated that MV regulated the differentiation and proliferation of diaphragm satellite cells.

MicroRNAs(miRNAs)是一组高度保守的非蛋白编码核苷酸序列,长度约为20～22个核苷酸.miRNAs最早在线虫体内被发现,此后,对miRNAs的功能及作用机制的研究逐渐成为热点.目前,研究表明miRNAs参与肺的生长发育、炎性反应及免疫调控等,与呼吸系统疾病的发生、发展及转归有着密切的联系.明确miRNAs的作用靶点、运用miRNA干扰技术探索相关信号转导通路将有助于加深对相关疾病的理解,为临床诊治提供新的依据和方向.本文将对miRNAs的研究背景、形成机制、生物学功能,以及在各种呼吸系统炎性疾病中的研究及应用进展进行综述.以期为呼吸系统炎性疾病的诊治提供新的思路.

Ethnopharmacological relevance: Qiguiyin, a hospital preparation of traditional Chinese medicinal formula, is a combination of Astragalus hamosus L., Angelica sinensis (Oliv.) Diels, Lonicera sempervirens L., Artemisia annua L., and Polygonum cuspidatum Siebold & Zucc. at a ratio of 12:3:3:2:2. It has been used to treat severe pneu-monia caused by drug-resistant bacteria in clinical practice, while studies on its toxicological safety are rare in the literature. Aim of the study: In the present study, we aimed to develop a new application of Qiguiyin according to the general research routine of traditional Chinese medicine (TCM), and the toxicological effects of the Qiguiyin formula at the treatment phase and recovery phase were also evaluated. Materials and methods: The rats were administered with the Qiguiyin formula at 10, 30, and 50 times of the corresponding dosage in humans for 13 consecutive weeks. During 13 weeks of the treatment phase and 4 weeks of the recovery phase, the general signs of toxicity and mortality were monitored daily, and the body weight and food consumption were determined every week. Moreover, the hematology, biochemistry, urine, organ weights, and histopathology were analyzed, and the reproductive system was examined at the end of the treatment phase or recovery phase, respectively. Results: The toxicological results showed no deaths and no changes in general behavior. Moreover, there was no clinically significant effect of the Qiguiyin formula on body weight or food consumption in rats. Although the Qiguiyin formula resulted in some changes in hematological, biochemical, and urinary indexes, these alterations were not related to the treatment because they remained within normal ranges throughout the 17 weeks. Besides, the main organs were not affected basically. All the above-mentioned results showed no gender difference. Furthermore, a clinical dosage of 50 times of the Qiguiyin formula did not affect the reproductive system of female rats, while it could lead to atrophied seminiferous tubules in two out of 10 male rats. However, such abnormality could not be found at the end of the recovery phase. Conclusions: Overall, the Qiguiyin formula could be used safely. The administration at doses of less than 1000 g/ day for 13 weeks showed no distinct toxicity or side effects.

Objective: Antibiotic resistance of Pseudomonas aeruginosa (PA) that lowers the effectiveness of current treatments for pneumonia is a growing problem. Qi Gui Yin is a Chinese herbal medicine that has been used to improve the efficacy of antibiotic therapy against antibiotic-resistant bacteria. This study aimed to elucidate the mechanism by which Qi Gui Yin inhibits antibiotic resistance of PA. Methods: Active components of Qi Gui Yin were analyzed by chromatography. Isobaric Tags for Relative and Absolute Quantification (iTRAQ) technology was used to compare protein expression profiles of PA strains cultured in serum from rats that were and were not treated with Qi Gui Yin. Quantitative polymerase chain reaction (qPCR) analysis was performed to detect gene expression changes. Results: Proteomic analysis identified 76 differentially expressed proteins between PA strains cultured in serum from rats that were or were not treated with Qi Gui Yin. Bioinformatics analysis revealed that the largest number of differentially expressed proteins were associated with resistance mechanisms such as quorum sensing, bacterial biofilm formation, and active pumping. In addition, qPCR analysis confirmed that downregulation of iscU and arcA gene expression was associated with Qi Gui Yin treatment. Conclusions: Serum from Qi Gui Yin-treated rats could effectively inhibit antibiotic resistance of PA. Chlorogenic acid and astragaloside IV are the main components of Qi Gui Yin, which may mediate inhibition of antibiotic resistance. Our findings provide new insights into strategies involving Chinese herbal medicine that can be used to treat pneumonia caused by antibiotic-resistant bacteria.

Gram-negative bacteria (GNB) emerge as important pathogens causing pulmonary infection, which can develop into sepsis due to bacterial resistance to antibiotics. GNB pneumonia poses a huge social and economic burden all over the world. During GNB infection in the lung, Toll-like receptor 4 (TLR4) can form a complex with MD2 and CD14 after recognizing lipopolysaccharide of GNB, initiate the MyD88- and TRIF-dependent signalling pathways and stimulate host non-specific immune response. In this review, we summarize recent progress in our understanding of the role of TLR4 in GNB pneumonia. The latest experimental results, especially in TLR4 knockout animals, suggest a promising potential of targeting TLR4 signalling pathway for the treatment of GNB pneumonia. Furthermore, we highlight the benefits of Traditional Chinese Medicine as novel candidates for the therapy of GNB pneumonia due to the modulation of TLR4 signalling pathway. Finally, we discuss the promise and challenge in the development of TLR4-based drugs for GNB pneumonia.

目的:客观评价已建立的滴鼻途径感染致肺炎大鼠模型,为后续肺炎防治机制研究提供实验依据.方法:将24只SD雄性大鼠按随机数表法分为对照组和实验组,每组12只,两组均按干预后0 d、5 d、10 d和15 d分为4个亚组,每组3只.以滴鼻途径干预,采集干预0 d、5 d、10 d和15 d的各亚组肺泡灌洗液(BALF)及肺组织样本,检测其肺指数、BALF中白细胞数、免疫炎性因子IL-4、TNF-α和IFN-γ的表达及铜绿假单胞菌数量,并分别与对照组的相应亚组比较分析.结果:实验组大鼠肺指数在造模干预后,随时间延长5 d、10 d和15 d逐渐增大,与对照组相比差异有统计学意义(t=-2.46,t=-3.26,t=-3.73;P＜0.05);BALF中白细胞数均明显高于相应对照亚组,BALF中IL-4和TNF-α在造模干预后均呈增长趋势,两组比较差异有统计学意义(t=4.24,t=4.88;P＜0.05),IFN-γ则呈下降趋势;BALF中铜绿假单胞菌数在造模干预后,随时间延长呈增多趋势.结论:成功构建的滴鼻感染耐药铜绿假单胞菌致大鼠肺炎模型,可用于中医药防治肺炎的机制研究.

目的:初步探索芪归银对耐药铜绿假单胞菌(PA)致肺炎大鼠的免疫影响,为全面揭示其疗效机制奠定基础.方法:在空白和模型对照基础上,设置芪归银实验组与阿莫西林西药干预对照组.采集各组肺组织,并腹主动脉采血分离血清.检测肺组织中高迁移率族蛋白B1(HMGB1)的蛋白表达,血清中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的浓度,进一步深入检测肺组织中HMGB1及其受体晚期糖基化终末产物受体(RAGE)、Toll-样受体4(TLR4)的基因表达.结果:模型组大鼠肺组织HMGB1蛋白及基因表达较对照组均明显升高,芪归银可一定程度降低其蛋白和基因表达;但阿莫西林对HMGB1和受体RAGE、TLR4的基因表达影响更为显著.模型组大鼠血清中IL-1β和TNF-α浓度较对照组均明显升高,芪归银可降低二者浓度,其中,对TNF-α浓度影响较阿莫西林更为显著.结论:肺炎发生时,芪归银对肌体核心免疫网络因子HMGB1及其受体RAGE、TLR4表达均有影响,对重要信号通路效应分子IL-1β和TNF-α也有影响,更深入的疗效机制有待进一步研究揭示.

目的 检测低浓度左氧氟沙星对铜绿假单胞菌(PA)感染小鼠肺炎模型的影响,探索左氧氟沙星耐药产生的方法.方法 采用PA感染小鼠致肺炎模型,各实验中小鼠按体重随机分组,以不同的菌液浓度、感染次数、给药方法,以小鼠体重变化规律、小鼠肺指数为评价指标,分析左氧氟沙星的治疗效果,探讨模型建立的可行性.结果低浓度左氧氟沙星对PA(1×109cfu/mL)单次感染小鼠,模型组1、模型组2肺指数与正常组比较均有显著性差异(P<0.01);左氧氟沙星组2与模型组2比较无显著性差异.低浓度左氧氟沙星对PA(1×1010cfu/mL)单次感染小鼠,模型组1和模型组2与正常组比较,均有显著性差异(P<0.01).左氧氟沙星组3与模型组2比较,结果无显著性差异.左氧氟沙星组1、2均能显著性降低模型组肺指数;PA重复感染小鼠实验中,模型组1和模型组2与正常组比较,均有显著性差异(P<0.01).左氧氟沙星组2与模型组2比较,无显著性差异.各实验中,小鼠感染后第二天,体重较正常组降低,结果无显著性差异.结论 实验条件下,低浓度左氧氟沙星对PA(1×1010cfu/mL)单次感染小鼠、低浓度左氧氟沙星对PA重复感染小鼠中,动物模型可在一定程度上模拟临床中耐药产生的现象.

脓毒症高致死率使其成为人类健康的重大威胁,其发病机制尚未完全明确,已知免疫失衡是其主要发病机制之一.高迁移率族蛋白B1(High Mobility Group Box1 Protein,HMGB1)是致免疫失衡的关键因子,有RAGE、TLRs和CD24等多种受体,可通过与受体结合,介导多种信号转导通路,从而发挥免疫调节功能.中药以单体和复方治疗脓毒症均取得一定进展,有待进一步结合现代医学共同应对脓毒症挑战.

目的:探讨芪归银方对铜绿假单胞菌的作用,对比评价该方水提醇沉与水提醇提提取物的有效性.方法:采用小鼠感染铜绿假单胞菌致肺炎模型,观察苠归银方提取物对该模型小鼠的治疗作用;采用肉汤二倍稀释法,检测芪归银方体外干预细菌生长的作用.结果:芪归银方两种提取物均可明显降低耐药铜绿假单胞菌致小鼠感染肺炎模型的肺指数,与耐药菌株模型对照组比较有显著性差异(P＜0.01);在同等剂量下,芪归银方水提醇沉提取物对该模型小鼠肺指数抑制率高于水提醇提组;体外对菌株生长均有不同程度的干预作用,其中水提醇沉与水提醇提提取物对菌株PA-S的MIC、MBC值分别为0.50g/mL、＞1.00g/mL,0.51g/mL、＞1.00g/mL;对菌株PA-R的MIC、MBC值分别为0.25、0.50g/mL,0.25、0.51g/mL.结论:芪归银方对铜绿假单胞菌有一定的干预作用,且水提醇沉提取物的作用优于水提醇提提取物.

Objective To explore the preparation of a rat model of pneumonia model induced by Pseudomonas aeruginosa( PA) using different methods,and to lay the foundation for further studies. Methods 48 SD rats were randomly divided into 4 groups:the control group (A), the intratracheal injection group (B), the trachea cannulation group (C) and the intranasal inoculation group ( D) . After intervention with different treatment modalities, the body weight,tempera-ture,white blood cell count and lung pathological changes in the rats of all groups were detected at 5, 10, 15 days. Results 1. The behavior, body weight, temperature, leukocytes and pathological inflammatory changes of the lung in rats of the model groups were significantly different from that of control group. 2. Pseudomonas aeruginosa was detected in rats of all the model groups, but the control group was negative. Conclusions Rat model of Pseudomonas aeruginosa infected pneu?monia can be successfully established by intranasal inoculation. This method can avoid the inflammatory interference from operation, and is simple and suitable for popularization.

Objective To investigate the clinical distribution and drug resistance characteristics of Klebsiella pneumonia in different hospitals,to analyze KPC gene,and to provide the basis for rational drug selection.Methods The pathogenic bacteria were isolated from clinical samples and identified by the two hospitals in 2015 respectively.The sample characteristics in clinical departments and type distribution were investigated.The results about drug resistance were verified with K-B method.All the Klebsiella pneumonia resistant to imipenem and meropenem were selected,of which the KPC genes were detected by PCR.Results The 5 075 and 1 694 strains bacteria were isolated from two hospitals respectively,including 524 strains Klebsiella pneumoniae and 286 strains respectively,which distributed mainly in ICU and respiratory department and from sputum,urine and blood samples.The drug resistance of Klebsiella pneumoniae in Tianjin Fourth Hospital were compound sulfamethoxazole,ampicillin,nitrofurantoin,ampicillin / sulbactam and piperacillin in order,while those in the other hospital were compound sulfamethoxazole,ampicillin,nitrofurantoin,ampicillin / sulbactam and cefazolin orderly.Some Klebsiella pneumoniae resistant to imipenem and meropenem could express the KPC gene.Conclusion The distribution characteristics and sample types of the two hospitals were similar,and the drug resistance spectrum was slightly different.These indicated that the respiratory tract infection in the respiration department and ICU should be paid attention to,and the rational drug was selected according to the drug resistance gene expression.

Interleukin 15 (IL-15) is a cytokine exhibiting antitumor characteristic similar to that of IL-2. However, in human tissues and cells, IL-15 expression and secretion is very limited, suggesting IL-15 functions mainly intracellularly. In the present study, we assessed the effects of transfecting NCI-H446 small cell lung cancer cells with genes encoding three IL-15 variants: prototypical IL-15, mature IL-15 peptide, and modified IL-15 in which the IL-2 signal peptide is substituted for the native signal peptide. NCI-H446 cells transfected with empty plasmid served as the control group. We found that IL-15 transfection effectively inhibited NCI-H446 cell proliferation and arrested cell cycle progression, with the modified IL-15 carrying the IL-2 signal peptide exerting the greatest effect. Consistent with those findings, expression each of the three IL-15 variants reduced growth of NCI-H446 xenograph tumors, and the modified IL-15 again showed the greatest effect. In addition, IL-15 expression led to down-regulation of the positive cell cycle regulators cyclin E and CDK2 and up-regulation of the negative cycle regulators p21 and Rb. These findings suggest IL-15 acts as a tumor suppressor that inhibits tumor cell proliferation by inducing cell cycle arrest.