ß-sitosterol ameliorates influenza A virus-induced proinflammatory response and acute lung injury in mice by disrupting the cross-talk between RIG-I and IFN/STAT signaling.

ß-Sitosterol (24-ethyl-5-cholestene-3-ol) is a common phytosterol Chinese medical plants that has been shown to possess antioxidant and anti-inflammatory activity. In this study we investigated the effects of ß-sitosterol on influenza virus-induced inflammation and acute lung injury and the molecular mechanisms. We demonstrate that ß-sitosterol (150-450 µg/mL) dose-dependently suppresses inflammatory response through NF-κB and p38 mitogen-activated protein kinase (MAPK) signaling in influenza A virus (IAV)-infected cells, which was accompanied by decreased induction of interferons (IFNs) (including Type I and III IFN). Furthermore, we revealed that the anti-inflammatory effect of ß-sitosterol resulted from its inhibitory effect on retinoic acid-inducible gene I (RIG-I) signaling, led to decreased STAT1 signaling, thus affecting the transcriptional activity of ISGF3 (interferon-stimulated gene factor 3) complexes and resulting in abrogation of the IAV-induced proinflammatory amplification effect in IFN-sensitized cells. Moreover, ß-sitosterol treatment attenuated RIG-I-mediated apoptotic injury of alveolar epithelial cells (AEC) via downregulation of pro-apoptotic factors. In a mouse model of influenza, pre-administration of ß-sitosterol (50, 200 mg·kg-1·d-1, i.g., for 2 days) dose-dependently ameliorated IAV-mediated recruitment of pathogenic cytotoxic T cells and immune dysregulation. In addition, pre-administration of ß-sitosterol protected mice from lethal IAV infection. Our data suggest that ß-sitosterol blocks the immune response mediated by RIG-I signaling and deleterious IFN production, providing a potential benefit for the treatment of influenza.

A novel allosteric site in casein kinase 2α discovered using combining bioinformatics and biochemistry methods.

Casein kinase 2 (CK2) is a highly pleiotropic serine-threonine kinase, which catalyzed phosphorylation of more than 300 proteins that are implicated in regulation of many cellular functions, such as signal transduction, transcriptional control, apoptosis and the cell cycle. On the other hand, CK2 is abnormally elevated in a variety of tumors, and is considered as a promising therapeutic target. The currently available ATP-competitive CK2 inhibitors, however, lack selectivity, which has impeded their development in cancer therapy. Because allosteric inhibitors can avoid the shortcomings of conventional kinase inhibitors, this study was aimed to discover a new allosteric site in CK2α and to investigate the effects of mutations in this site on the activity of CK2α. Using Allosite based on protein dynamics and structural alignment, we predicted a new allosteric site that was partly located in the αC helix of CK2α. Five residues exposed on the surface of this site were mutated to validate the prediction. Kinetic analyses were performed using a luminescent ADP detection assay by varying the concentrations of a peptide substrate, and the results showed that the mutations I78C and I78W decreased CK2α activity, whereas V31R, K75E, I82C and P109C increased CK2α activity. Potential allosteric pathways were identified using the Monte Carlo path generation approach, and the results of these predicted allosteric pathways were consistent with the mutation analysis. Multiple sequence alignments of CK2α with the other kinases in the family were conducted using the ClustalX method, which revealed the diversity of the residues in the site. In conclusion, we identified a new allosteric site in CK2α that can be altered to modulate the activity of the kinase. Because of the high diversity of the residues in the site, the site can be targeted using rational drug design of specific CK2α inhibitors for biological relevance.

Simvastatin downregulates expression of TGF-ßRII and inhibits proliferation of A549 cells via ERK.

Lung cancer is the leading cause of cancer-related death worldwide. Transforming growth factor-ß receptor II (TGF-ßRII) plays an important role in the regulation of proliferation and progression in cancer. Statins have been documented to exhibit anticancer and cancer chemopreventive properties. However, the effects and mechanisms of simvastatin on the development of lung cancer are still unclear. In the present study, quiescent A549 cells were treated in vitro with fetal bovine serum (FBS) in the presence or absence of simvastatin. MTT, Western blot, and real-time qPCR were used to detect cell viability, activation of ERK, and expression of TGF-ßRII at the protein and RNA level. Our results demonstrated that simvastatin inhibited activation of ERK, downregulated expression of TGF-ßRII, and suppressed A549 cell proliferation. Furthermore, the effects of simvastatin can be reversed by farnesyl pyrophosphate (FPP). Therefore, these results suggest that simvastatin may inhibit A549 cell proliferation and downregulate TGF-ßRII expression by inhibiting activation of ERK. Our findings may advance the current understanding of the effects of simvastatin on cancer progression and contribute to the study of cancer treatment.

Comparison of the outcomes of monopolar and bipolar radiofrequency ablation in surgical treatment of atrial fibrillation.

OBJECTIVE: To compare the therapeutic effects and safety of monopolar and bipolar radiofrequency (RF) ablation used during cardiac surgery to treat atrial fibrillation. METHODS: We retrospectively studied a total of 81 patients with chronic atrial fibrillation who underwent open cardiac surgery with concomitant RF ablation between January 2007 and March 2011. Fifty-eight patients received bipolar RF ablation and 23 received monopolar RF ablation, respectively. The sinus rhythm restoration rate, the procedural duration, the frequency of severe perioperative complications, and mortality were compared between the two groups. RESULTS: The sinus rhythm restoration rate did not differ significantly between the two groups after follow-up of 15.1 ± 12.6 months (P=0.199). The frequencies of severe perioperative complications and mortality were also similar in the two groups. The total procedural time using bipolar RF ablation was significantly shorter than that using monopolar ablation (19.7 ± 4.6 minutes vs. 28.1 ± 8.5 minutes, P< 0.001). CONCLUSIONS: Both monopolar and bipolar RF ablation are safe and effective in treating chronic atrial fibrillation patients during open cardiac surgery, but bipolar RF ablation is more convenient in practice.

[Clinical application of extracorporeal membrane oxygenation for treatment of adult refractory cardiogenic shock].

OBJECTIVE: To summarize the clinical experience of extracorporeal membrane oxygenation (ECMO) treatment for adult refractory cardiogenic shock. METHODS: From January 2003 to January 2011, patients with refractory cardiogenic shock required veno-arterial ECMO by failure of conventional therapy and intra-aortic balloon pump counterpulsation therapy were retrospectively studied. Patients with severe traumatic brain injury, advanced malignancies and multiple organ failure were excluded. Patients were divided into weaned group (n = 31) and not weaned group (n = 23) according to the ECMO weaning. RESULTS: The duration of ECMO was 24.16 (14.12, 56.75) hours. Twenty-two out of 31 patients in the weaned group survived and were discharged, 9 patients died after successfully weaned from ECMO (5 due to multisystem organ failure, 2 due to reoccurred cardiogenic shock, 1 due to infectious shock and 1 due to disseminated or diffuse intravascular coagulation). Pre-ECMO mean arterial pressure, ejection fraction, the duration of ECMO were significantly higher while pre-ECMO blood lactate [(8.64 ± 3.17) vs. (14.44 ± 2.52) , P < 0.01], the duration of ROSC [ (16.70 ± 5.29) vs. (35.64 ± 5.89), P < 0.01] and multisystem organ failure [0 vs. 17.4% (4/23) , P < 0.05] were lower in weaned group than in not wean group. CONCLUSIONS: ECMO is an effective mechanical assistant therapy strategy for adult refractory cardiogenic shock patients. Timely applying this strategy on suitable patients is crucial for the success of ECMO. Cardiac function and reversibility of heart failure are key factors determine the fate of weaned or not weaned ECMO in adult refractory cardiogenic shock patients.

Coinfection with influenza virus and non-typeable Haemophilus influenzae aggregates inflammatory lung injury and alters gut microbiota in COPD mice.

Background: Acute exacerbation of chronic obstructive pulmonary disease (AECOPD) is associated with high mortality rates. Viral and bacterial coinfection is the primary cause of AECOPD. How coinfection with these microbes influences host inflammatory response and the gut microbiota composition is not entirely understood. Methods: We developed a mouse model of AECOPD by cigarette smoke exposure and sequential infection with influenza H1N1 virus and non-typeable Haemophilus influenzae (NTHi). Viral and bacterial titer was determined using MDCK cells and chocolate agar plates, respectively. The levels of cytokines, adhesion molecules, and inflammatory cells in the lungs were measured using Bio-Plex and flow cytometry assays. Gut microbiota was analyzed using 16S rRNA gene sequencing. Correlations between cytokines and gut microbiota were determined using Spearman's rank correlation coefficient test. Results: Coinfection with H1N1 and NTHi resulted in more severe lung injury, higher mortality, declined lung function in COPD mice. H1N1 enhanced NTHi growth in the lungs, but NTHi had no effect on H1N1. In addition, coinfection increased the levels of cytokines and adhesion molecules, as well as immune cells including total and M1 macrophages, neutrophils, monocytes, NK cells, and CD4 + T cells. In contrast, alveolar macrophages were depleted. Furthermore, coinfection caused a decline in the diversity of gut bacteria. Muribaculaceae, Lactobacillus, Akkermansia, Lachnospiraceae, and Rikenella were further found to be negatively correlated with cytokine levels, whereas Bacteroides was positively correlated. Conclusion: Coinfection with H1N1 and NTHi causes a deterioration in COPD mice due to increased lung inflammation, which is correlated with dysbiosis of the gut microbiota.

Personalized management of anastomotic leak after surgery for esophageal carcinoma.

OBJECTIVE: To summarize the management of anastomotic leak following surgery for esophageal carcinoma. METHODS: The medical records of the patients developing digestive tract leak after surgery for esophageal carcinoma in our hospital from January 2003 to March 2011 were retrospectively analyzed. RESULTS: A total of 36 patients were included, in whom 13 developed cervical anastomotic leak, 18 had intra-thoracic anastomotic leak, and 5 had intra-thoracic gastric necrosis. Of these patients, 7 were treated with resurgery, 6 with esophageal stent implantation, and 23 with conservative treatment. Treatment lasted for 5 to 181 days, averagely 47.0 +/- 31.9 days. After management, 9 patients died (25.0%). Among seven patients with resurgery, four had deceased, two were cured, and one developed leak again and was switched to conservative treatment until discharged. All the 6 patients treated with stent implantation were cured. Of the 24 patients receiving conservative treatment (including one switched from resurgery), 18 (75.0%) were cured and 1 was not cured but survived. CONCLUSIONS: Anastomotic leak following surgery for esophageal carcinoma should be treated individually based on the onset time, location, size, and extent of the leakage. Conservative treatment is still a safe and effective method. The efficacy of stent implantation needs further investigation to confirm.

[Research advances in direct interspecies electron transfer within microbes]. / å¾®ç”Ÿç‰©ç§é—´ç›´æŽ¥ç”µå­ä¼ é€’ç ”ç©¶è¿›å±•.

Hydrogen and formic acid have been considered as the intermediate electron transporters among microbes for a long time. In recent years, however, it has been found that direct interspecies electron transfer (DIET) might be an alternative beyond hydrogen/formic acid to transfer electron among microbes. As a new way of electron transfer among microbes, the electron transfer efficiency of DIET is higher than that of traditional hydrogen/formate transfer. The discovery of DIET has changed the traditional understanding that the growth and metabolism of microbial syntrophism must rely on electron carriers such as hydrogen or formic acid, and also has opened a new perspective for the study of microbial interaction. Although great progress has been made in the study of DIET, in-depth studies are still lacking on the microbes that can form co-culture via DIET, the mechanism of DIET, and the factors affecting DIET. In this review, we summarized the microbes that can form DIET, the mechanism underlying the extracellular electron transfer of microbe acted as electron donor in DIET, as well as the mechanism underlying the extracellular electron transfer of microbe acted as electron acceptor in DIET. The effects of conductive materials on DIET were elaborated, and several research directions for DIET were proposed, with the aim to mitigate performance degradation and facilitate research and development in this area.

Pathologic complete response after neoadjuvant tislelizumab and chemotherapy for Pancoast tumor: A case report.

A 60-year-old man was hospitalized because of numbness and weakness in the right upper limb. Magnetic resonance imaging revealed a large mass in the right upper lobe invading the right eighth cervical and first thoracic nerve root. Biopsy pathology confirmed primary lung adenocarcinoma with a clinical stage of cT4N0M0 IIIA, negative for anaplastic lymphoma kinase fusion gene and epidermal growth factor receptor mutations but positive for programmed death ligand 1 (3%). Neoadjuvant tislelizumab and chemotherapy were offered to this patient with Pancoast tumor, and tumor shrinkage of 71% was achieved. After the operation, surgical pathology indicated pathologic complete response (pCR). Circulating tumor cells testing was negative after the first adjuvant treatment. In this case, we provide real-world evidence of encouraging pCR with neoadjuvant tislelizumab and chemotherapy for a patient with Pancoast tumor.

Novel Fatty Acid in Cordyceps Suppresses Influenza A (H1N1) Virus-Induced Proinflammatory Response Through Regulating Innate Signaling Pathways.

Influenza virus (IV) infections usually cause acute lung injury characterized by exaggerated proinflammatory responses. The paucity of therapeutic strategies that target host immune response to attenuate lung injury poses a substantial challenge in management of IV infections. In this study, we chemically synthesized a novel fatty acid (2Z,4E)-deca-2,4-dienoic acid (DDEA) identified from Chinese Cordyceps by using UHPLC-Q-TOF-MS techniques. The DDEA did not inhibit H1N1 virus replication but attenuated proinflammatory responses by reducing mRNA and protein levels of TNF-α, IFN-α, IFN-ß, IL-6, CXCL-8/IL-8, CCL-2/MCP-1, CXCL-10/IP-10, CCL-3/MIP-1α, and CCL-4/MIP-1ß in A549 cells and U937-derived macrophages. The anti-inflammatory effect occurred through downregulations of TLR-3-, RIG-I-, and type I IFN-activated innate immune signaling pathways. Altogether, our results indicate that DDEA may potentially be used as an anti-inflammatory therapy for the treatment of IV infections.

Pump scheme for gain-flattened Raman fiber amplifiers using improved particle swarm optimization and modified shooting algorithm.

An effective pump scheme for the design of broadband and flat gain spectrum Raman fiber amplifiers is proposed. This novel approach uses a new shooting algorithm based on a modified Newton-Raphson method and a contraction factor to solve the two point boundary problems of Raman coupled equations more stably and efficiently. In combination with an improved particle swarm optimization method, which improves the efficiency and convergence rate by introducing a new parameter called velocity acceptability probability, this scheme optimizes the wavelengths and power levels for the pumps quickly and accurately. Several broadband Raman fiber amplifiers in C+L band with optimized pump parameters are designed. An amplifier of 4 pumps is designed to deliver an average on-off gain of 13.3 dB for a bandwidth of 80 nm, with about +/-0.5 dB in band maximum gain ripples.

µ-Cyanido-1:2κN:C-tricyanido-2κC-(rac-5,5,7,12,12,14-hexa-methyl-1,4,8,11-tetra-aza-cyclo-tetra-decane-1κN,N',N'',N''')dinickel(II) N,N-di-methyl-formamide monosolvate hemi-hydrate.

The two Ni(II) atoms in the title complex, [Ni(2)(CN)(4)(C(16)H(36)N(4))]·C(3)H(7)NO·0.5H(2)O, are bridged by a cyanide ion. The macrocycle folds around one Ni(II) atom, which is five-coordinated in an NiN(5) square-pyramidal geometry. The other Ni(II) atom is surrounded by the cyanide ions in an NiN(4) square-planar geometry. The dimethyl-formamide solvent mol-ecule is disordered over two positions in a 0.62 (1):0.38 (1) ratio and the water mol-ecule is disordered about a center of inversion. The dinuclear mol-ecule and solvent mol-ecules are linked by N-H⋯O, N-H⋯N and O-H⋯O hydrogen bonds, forming a three-dimensional network.

Total alkaloids from Alstonia scholaris inhibit influenza a virus replication and lung immunopathology by regulating the innate immune response.

BACKGROUND: Alstonia scholaris is a folk medicine used to treat cough, asthma and chronic obstructive pulmonary disease in China. Total alkaloids (TA) from A. scholaris exhibit anti-inflammatory properties in acute respiratory disease, which suggests their possible anti-inflammatory effect on influenza virus infection. PURPOSE: To assess the clinical use of TA by demonstrating their anti-influenza and anti-inflammatory effects and the possible mechanism underlying the effect of TA on influenza A virus (IAV) infection in vitro and to reveal the inhibitory effect of TA on lung immunopathology caused by IAV infection. METHODS: Antiviral and anti-inflammatory activities were assessed in Madin-Darby canine kidney (MDCK) and A549 cells and U937-derived macrophages infected with influenza A/PR/8/34 (H1N1) virus. Proinflammatory cytokine levels were measured by real-time quantitative PCR and Bio-Plex assays. The activation of innate immune signaling induced by H1N1 virus in the absence or presence of TA was detected in A549 cells by Western blot. Furthermore, mice were infected intranasally with H1N1 virus and treated with TA (50, 25 and 12.5 mg/kg/d) or oseltamivir (60 mg/kg/d) for 5 days in vivo. The survival rates and body weight were recorded, and the viral titer, proinflammatory cytokine levels, innate immune cell populations and histopathological changes in the lungs were analyzed. RESULTS: TA significantly inhibited viral replication in A549 cells and U937-derived macrophages and markedly reduced cytokine and chemokine production at the mRNA and protein levels. Furthermore, TA blocked the activation of pattern recognition receptor (PRR)- and IFN-activated signal transduction in A549 cells. Critically, TA also increased the survival rate, reduced the viral titer, suppressed proinflammatory cytokine production and innate immune cell infiltration and improved lung histopathology in a lethal PR8 mouse model. CONCLUSION: TA exhibits anti-viral and anti-inflammatory effects against IAV infection by interfering with PRR- and IFN-activated signal transduction.

[bcl-2 expression in small cell lung cancer: a mechanism for apoptosis antagonism and immune evasion].

OBJECTIVE: To observe bcl-2 expression and its correlation with apoptosis antagonism and immune evasion of small cell lung cancer cell line and explore the application of bcl-2 antisense thio-oligonucleotide (bcl-2 SON) in gene therapy for lung cancer. METHODS: Western blotting was performed to detect bcl-2 expression in NCI-H69 cell line expressing Bcl-2 protein treated with bcl-2 SON. The treated NCI-H69 cells, along with the cells without bcl-2 SON treatment and NCI-H82 cells that did not express Bcl-2 protein, were respectively co-cultured with tumor-infiltrating lymphocytes (TILs) isolated from fresh tumor samples, and the apoptosis of the lung cancer cells was assessed by JAM assay. RESULTS: Western blotting revealed obvious inhibition of bcl-2 expression in NCI-H69 cells in response to bcl-2 SON treatment. JAM assay showed that the apoptosis of bcl-2 SON-treated H69 cells and NCI-H82 cells increased with the elevation of TIL ratio in the co-culture, while such changes were not observed in NCI-H69 cells without bcl-2 SON treatment. CONCLUSION: Small cell lung cancer cells expressing bcl-2 may antagonize the antitumor immune attack in the host, and bcl-2 SON may provide an effective alternative in gene therapy for small cell lung cancer.