Anti-inflammatory effects of steroidal alkaloids of Solanum lyratum Thunb.

Solanum lyratum Thunb., a traditional Chinese herbal medicine, has a promising background. However, the anti-inflammatory effects of its component steroid alkaloid have not been explored. In this study, animal and cell experiments were performed to investigate the anti-inflammatory effects and mechanism of action of Solanum lyratum Thunb steroid alkaloid (SLTSA), in order to provide evidence for its potential utilization. SLTSA effectively inhibited ear swelling and acute abdominal inflammation of mice. We observed concentration-dependent inhibition of pro-inflammatory cytokines by SLTSA, as confirmed by the ELISA and RT-qPCR results. Flow cytometry, immunofluorescence and RT-qPCR analyses revealed that SLTSA suppressed TLR4 expression. Western blot results indicated that SLTSA inhibited the activation of the TLR4/MyD88/NF-κB signaling pathway. Our study demonstrated that SLTSA possesses anti-inflammatory properties.

IFN-γ-/- Mice Resist Actinobacillus pleuropneumoniae Infection by Promoting Early Lung IL-18 Release and PMN-I Accumulation.

Porcine pleuropneumonia is a common infectious disease of pigs caused by Actinobacillus pleuropneumoniae Interferon gamma (IFN-γ) expression increases in the lung of pigs after A. pleuropneumoniae infection, but the role of IFN-γ during the infection is still obscure. In this study, an IFN-γ-/- mouse infection model was established, and bacterial load, levels of inflammatory cytokines, and types of neutrophils in the lungs were studied at different times post-A. pleuropneumoniae infection. We found that wild-type (WT) mice were more susceptible to A. pleuropneumoniae than IFN-γ-/- mice. At 6 h postinfection (hpi), the expression of interleukin 18 (IL-18) and IL-1ß in the lungs of IFN-γ-/- mice was significantly increased compared to WT mice. The bacterial load and levels of inflammatory cytokines (IL-1ß and IL-6) of IFN-γ-/- mice were significantly reduced at 12 hpi compared to WT mice. After an initial loss, the numbers of lung polymorphonuclear (PMN)-I cells dramatically increased in the lungs of IFN-γ-/- but not WT mice, whereas PMN-II cells continually decreased. Finally, in vivo administration of IL-18 significantly reduced clinical scores and bacterial load in the lungs of A. pleuropneumoniae-infected mice. This study identifies IFN-γ as a target for regulating the inflammatory response in the lung and provides a basis for understanding the course of clinical bacterial pneumonia and for the formulation of treatment protocols.

Identification of HMMR as a prognostic biomarker for patients with lung adenocarcinoma via integrated bioinformatics analysis.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most prevalent tumor in lung carcinoma cases and threatens human life seriously worldwide. Here we attempt to identify a prognostic biomarker and potential therapeutic target for LUAD patients. METHODS: Differentially expressed genes (DEGs) shared by GSE18842, GSE75037, GSE101929 and GSE19188 profiles were determined and used for protein-protein interaction analysis, enrichment analysis and clinical correlation analysis to search for the core gene, whose expression was further validated in multiple databases and LUAD cells (A549 and PC-9) by quantitative real-time PCR (qRT-PCR) and western blot analyses. Its prognostic value was estimated using the Kaplan-Meier method, meta-analysis and Cox regression analysis based on the Cancer Genome Atlas (TCGA) dataset and co-expression analysis was conducted using the Oncomine database. Gene Set Enrichment Analysis (GSEA) was performed to illuminate the potential functions of the core gene. RESULTS: A total of 115 shared DEGs were found, of which 24 DEGs were identified as candidate hub genes with potential functions associated with cell cycle and FOXM1 transcription factor network. Among these candidates, HMMR was identified as the core gene, which was highly expressed in LUAD as verified by multiple datasets and cell samples. Besides, high HMMR expression was found to independently predict poor survival in patients with LUAD. Co-expression analysis showed that HMMR was closely related to FOXM1 and was mainly involved in cell cycle as suggested by GSEA. CONCLUSION: HMMR might be served as an independent prognostic biomarker for LUAD patients, which needs further validation in subsequent studies.

Comparison of in vivo versus in vitro fabrication of bone cement spacers in Masquelet technique for bone defects / 中华创伤骨科杂志

Objective:To compare in vivo versus in vitro fabrication of bone cement spacers in the treatment of bone defects by Masquelet technique. Methods:The data of 128 patients were analyzed retrospectively who had been treated for bone defects by Masquelet technique at Department of Orthopedics, Wuxi No. 9 People’s Hospital from January to August 2019. They were 74 males and 54 females, aged from 13 to 77 years. Their bone defects were traumatic in 54 cases and infectious in 74 cases. In 76 of them ( in vivo group), after a bone cement spacer was implanted into a bone defect during its dough phase, it was fabricated in vivo to form a cylindrical structure which was as large as or slightly larger than the defect size. In the other 52 cases ( in vitro group), before a bone cement spacer was implanted into a bone defect, it was fabricated in vivo during its dough phase into a cylindrical or block or bead chain or spherical form which was naturally solidificated at room temperature. The 2 groups were compared in terms of spacer filling time, bone healing time, delayed healing rate, infection control rate, spacer removal time, incidence of induced membrane or broken end bone lesion, as well as upper limb function evaluated by the Disability of the Arm, Shoulder and Hand Questionnaire (DASH) and the Paley lower limb grading at the last follow-up. Results:The 2 groups were comparable because there was no significant difference between them in gender, age, ratio of infected to non-infected cases, combined injuries, comorbidities or number of operations ( P>0.05). All the patients were followed up for 12 to 50 months (mean, 18.6 months). There were no significant differences between the 2 groups in spacer filling time, bone healing time, delayed healing rate, infection control rate or functional recovery for upper or lower limbs or for large or small bone defects (all P>0.05). In the in vivo group, for upper and lower limbs and for large and small bone defects respectively, the spacer removal time [(3.6±1.0) min, (4.1±1.1) min, (4.0±1.1) min and (3.9±1.0) min] and the incidence of induced membrane or broken end bone lesion [48.1%(13/27), 73.5%(36/49), 82.6%(39/46) and 66.7%(20/30)] were significantly longer or higher than those in the in vitro group [all (0.4±0.2) min; 3.2%(1/31), 9.5%(2/21), 0 (0/21) and 0 (0/31)] (all P<0.05). Conclusions:In the treatment of bone defects by Masquelet technique, in vivo and in vitro fabrication of bone cement spacers may lead to similar therapeutic effects. In vivo fabrication may be more suitable for lower limb, large or unstable bone defects but the spacer is not easy to remove and the induced membrane or bone ends are likely to get injured while in vitro fabrication may be more suitable for partial, small or upper limb defects because it may produce a variously shaped spacer.

The Shuganhuazheng Formula in Triple-Negative Breast Cancer: A Study Based on Network Pharmacology and In Vivo Experiments.

Breast cancer is the most common cancer in women. Among breast cancer subtypes, triple-negative breast cancer (TNBC) has the highest degree of malignancy and the worst prognosis. The Shuganhuazheng formula (SGHZF) is a traditional Chinese herbal formula for the treatment of TNBC, but the mechanism of SGHZF in the treatment of TNBC remains unclear. In this study, the therapeutic effect and mechanism of SGHZF against TNBC were preliminarily determined based on in vivo experimental verification and network pharmacology. In terms of therapeutic effects, the antitumour effect was verified by measuring and calculating tumour volume, and the expression of proto-oncogene c-Myc was verified by PCR. In terms of the mechanism, potential therapeutic targets were identified by overlapping the SGHZF-related and TNBC-related targets. After comprehensively analysing the results of the protein-protein interaction (PPI), gene ontology (GO) function, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, Akt and HIF-1α were selected for verification by using immunohistochemical and Western blot analyses. The results of the study indicated that SGHZF can inhibit breast tumour growth in mice and that the mechanism may be related to the inhibition of Akt and HIF-1α expression.

Systematic Evaluation of Efficacy and Safety of Acupuncture Treatment for Patients with Atrial Fibrillation.

BACKGROUND: Atrial fibrillation (AF) is one of the most common types of arrhythmia diagnosed in clinical practice. Due to its negative effects on people's physical and mental health, it is necessary to prevent and treat AF. Recently, scholars have found that acupuncture can be used to treat AF, but some scholars have questioned its therapeutic efficacy. AIM: Therefore, this study was performed to assess the efficacy and safety of acupuncture treatment for AF patients. METHODS: Previously published research articles were retrieved from six databases, and the data was analysed using RevMan5.3 software with a statistically significant difference defined as P < 0.05. RESULTS: A total of 8 relevant kinds of literature were retrieved containing 633 AF patients (323 in the treatment group and 310 in the control group). Acupuncture treatment increased the total efficacy and the rate of AF cardioversion to sinus rhythm (RR: 1.38; 95% CI: 1.25 to 1.53 vs RR: 1.40;95% CI: 1.16 to 1.69; each P < 0.05), and decreased the time of AF cardioversion to sinus rhythm, the heart rate and incidence of adverse effects (RR: -3.95; 95% CI: -4.98 to -2.91 vs RR: -14.54; 95% CI: -24.09 to -5.00 vs RR: 0.48; 95% CI: 0.21 to 1.11, each P < 0.05). There was difference between retention time more and less than 30 minutes (I2 = 74.9%, P = 0.05). The funnel plot displayed a symmetrical and funnel-form shape, indicating low bias. CONCLUSION: Acupuncture has a good therapeutic effect and safety profile on patients with AF, and its application in clinical practice should be considered.

Self-Assembled Ti4+@Biospore Microspheres for Sensitive DNA Analysis.

Ti4+ can be chemically adsorbed and assembled on the surface of the modified spore to form highly monodispersed Ti4+@spore microspheres. Moreover, we for the first time found that these biomicrospheres exhibit differential affinities toward ssDNA and dsDNA. As a principle-of-proof, we exploited the self-assembled Ti4+@spore microspheres for a hybridization analysis. Interestingly, in the hybridization analysis, residual ssDNA probes are selectively adsorbed on Ti4+@spore microspheres at pH 5.0 and then removed via centrifugation. By taking advantage of this property, the signal-to-noise ratio for DNA analysis was considerably increased by reducing the noise caused by the residual ssDNA probes. The proposed method features easy operation, high specificity, and sensitivity and thus exhibits potential for further applications on DNA biosensing.

Pnserpin: A Novel Serine Protease Inhibitor from Extremophile Pyrobaculum neutrophilum.

Serine protease inhibitors (serpins) are native inhibitors of serine proteases, constituting a large protein family with members spread over eukaryotes and prokaryotes. However, only very few prokaryotic serpins, especially from extremophiles, have been characterized to date. In this study, Pnserpin, a putative serine protease inhibitor from the thermophile Pyrobaculum neutrophilum, was overexpressed in Escherichia coli for purification and characterization. It irreversibly inhibits chymotrypsin-, trypsin-, elastase-, and subtilisin-like proteases in a temperature range from 20 to 100 °C in a concentration-dependent manner. The stoichiometry of inhibition (SI) of Pnserpin for proteases decreases as the temperature increases, indicating that the inhibitory activity of Pnserpin increases with the temperature. SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) showed that Pnserpin inhibits proteases by forming a SDS-resistant covalent complex. Homology modeling and molecular dynamic simulations predicted that Pnserpin can form a stable common serpin fold. Results of the present work will help in understanding the structural and functional characteristics of thermophilic serpin and will broaden the current knowledge about serpins from extremophiles.

Hydrogelation of the Short Self-Assembling Peptide I3QGK Regulated by Transglutaminase and Use for Rapid Hemostasis.

The self-assembly of short peptides is a promising route to the creation of smart biomaterials. To combine peptide self-assembly with enzymatic catalysis, we design an amphiphilic short peptide I3QGK that can self-assemble into long nanoribbons in aqueous solution. Upon addition of transglutaminase (TGase), the peptide solution undergoes a distinct sol-gel transition to form a rigid hydrogel, which shows strong shear-thinning and immediate recovery properties. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) measurements indicate the occurrence of considerable nanofibers in addition to the original nanoribbons. Liquid chromatography and mass spectrometry analyses reveal the enzymatic formation of peptide dimers from monomers through intermolecular ε-(γ-glutamyl)lysine isopeptide bonding. The dimers rapidly self-assemble into flexible and entangled nanofibers, and the coexistence of the original nanoribbons and the newly created nanofibers is responsible for hydrogelation. Factor XIII in blood is converted by thrombin to an active TGase (Factor XIIIa) during bleeding, so the peptide solution shows a more rapid and effective hemostasis via a combination of gelling blood and promoting platelet adhesion, relative to other hemostasis methods or materials. These features of I3QGK, together with its low cytotoxicity against normal mammalian cells and noninduction of nonspecific immunogenic responses, endow it with great potential for future clinical hemostasis applications.

PGBL inhibits the RAW 264.7 cells to express inflammatory factor.

Polysaccharides derived from Ginkgo biloba leaf (PGBL) is a kind of active ingredient came out from ginkgo biloba leaf extractions. Previous studies have shown that PGBL has a good anti-inflammatory effect. However, the mechanism is not clear. This study is to investigate the modulated immunity effect of PGBL on RAW264.7 cells. Here we showed that lipopolysaccharide (LPS) induces the expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and this induction can be repressed by PGBL treatment both in protein level and mRNA level, and PGBL strongly reduced the translocation of nuclear factor (NF)-κB to the cell nucleus. These findings demonstrate that PGBL can decrease the sensitivity of monocytes to LPS, and PGBL has applications in systemic inflammation and immune diseases.

Purified polysaccharide from Ginkgo biloba leaves inhibits P-selectin-mediated leucocyte adhesion and inflammation.

AIM: To investigate the anti-inflammatory mechanism of the polysaccharides of Ginkgo biloba leaves (PGBL) by inhibiting leucocyte adhesion. METHODS: The rough PGBL were isolated and purified. The anti-inflammatory effects of purified PGBL (p-PGBL) were assayed by ear edema induced by xylol and the acute peritonitis model in mice. The effect of p-PGBL on inhibiting the interaction between P-selectin and its ligands was investigated by flow cytometry and flow chamber. RESULTS: p-PGBL could effectively inhibit the acute inflammation in mice and interfere with the adhesion of HL-60 cells, a human leukaemia cell line, or neutrophils to P-selectin in static conditions, as well as the adhesion of neutrophils to Chinese hamster ovary cells expressing human P-selectin and human umbilical vein endothelial cells in flow conditions in a dose-dependant manner. CONCLUSIONS: p-PGBL can inhibit the inflammatory process through interfering with the interaction between P-selectin and its ligands.

P-Selectin-mediated acute inflammation can be blocked by chemically modified heparin, RO-heparin.

Selectins are carbohydrate-binding cell adhesion molecules that play a major role in the initiation of inflammatory responses. Heparin can bind to P-selectin, and its anti-inflammatory property is mainly due to inhibition of P-selectin. However, the strong anticoagulant activity of heparin limits its clinical use. We prepared periodate-oxidized, borohydride-reduced heparin (RO-heparin) by chemical modification and tested its anticoagulant and anti-inflammatory activities. Activated partial thromboplastin time (aPTT) assays showed that, compared with heparin, RO-heparin had greatly reduced anticoagulant activity. Intravenous administration of this compound led to reduction in the peritoneal infiltration of neutrophils in a mouse acute inflammation model. In vitro cell adhesion experiments demonstrated that the effect of RO-heparin on inflammatory responses was mainly due to inhibiting the interaction of P-selectin with its ligands. These results indicate that RO-heparin may be a safer treatment for inflammation than heparin, especially when selectin is targeted.