Increasing evidence has revealed that maternal cytomegalovirus (CMV) infection may be associated with neurodevelopmental disorders in offspring.Potential relevance between the placental inflammation and CMV-related autism has been reported by clinical observation.Meanwhile,abnormal expression of Toll-like receptor 2 (TLR2) and TLR4 in placenta of patients with chorioamnionitis was observed in multiple studies.IL-6 and IL-10 are two important maternal inflammatory mediators involved in neurodevelopmental disorders.To investigate whether murine CMV (MCMV) infection causes alterations in placental IL-6/10 and TLR2/4 levels,we analyzed the dynamic changes in gene expression of TLR2/4 and IL-6/10 in placentas following acute MCMV infection.Mouse model of acute MCMV infection during pregnancy was created,and pre-pregnant MCMV infected,lipopolysaccharide (LPS)-treated and uninfected mice were used as controls.At E13.5,E14.5 and E18.5,placentas and fetal brains were harvested and mRNA expression levels of placental TLR2/4 and IL-6/10 were analyzed.The results showed that after acute MCMV infection,the expression levels of placental TLR2/4 and IL-6 were elevated at E13.5,accompanied by obvious placental inflammation and reduction of placenta and fetal brain weights.However,LPS 50 μg/kg could decrease the IL-6 expression at E13.5 and E14.5.This suggests that acute MCMV infection during pregnancy could up-regulate the gene expression of TLR2/4 in placental trophoblasts and activate them to produce more proinflammatory cytokine IL-6.High dose of LPS stimulation (50 tg/kg) during pregnancy can lead to down-regulation of IL-6 levels in the late stage.Imbalance ofIL-6 expression in placenta might be associated with the neurodevelopmental disorders in progeny.

BACKGROUNDMurine cytomegalovirus (MCMV) early protein M112-113 is involved in viral DNA replication and believed to play a crucial role in the viral pathogenesis. To investigate the biological function of M112-113 protein in the pathogenesis of the brain disorders caused by cytomegalovirus (CMV), a screening for proteins interacting with M112-113 was performed by a yeast two-hybrid system.

METHODSBait plasmid pGBKT7-M112-113 was constructed and transformed into AH109 yeast. After confirmation of the expression of MCMV M112-113 in yeast, the bait yeast was mated with a prey yeast containing mouse brain cDNA library plasmid to screen the proteins interacting with M112-113. Interactions between M112-113 and the obtained proteins were verified by yeast two-hybrid assay and chemiluminescent co-immunoprecipitaion.

RESULTSTwo proteins interacting with M112-113 were identified, including metastasis-associated 1 (MTA1) and zinc finger, CCHC domain containing 18 (ZCCHC18). M112-113 protein could interact with MTA1 or ZCCHC18 in yeast and mammalian cells.

CONCLUSIONThe interactions of M112-113 with MTA1 or ZCCHC18 may be related to the pathogenesis of MCMV-associated disease in central nervous system.

Animals ; Brain ; metabolism ; Cell Line ; Humans ; Immunoprecipitation ; Mice ; Muromegalovirus ; metabolism ; Plasmids ; Protein Binding ; Two-Hybrid System Techniques ; Viral Proteins ; metabolism

Objective:To investigate the role of pro-inflammatory cytokine IL-17 involved in the pathogenesis of cytomegalovirus hepatitis in vivo.Methods:First of all,disseminated infection model was established.Then,mice were randomly divided into 4 groups:normal control group,MCMV-infected control group,IL-17 blockade group,and isotype control group.Mice were sacrificed on day 7 after infection.The levels of IL-17 protein were detected by Western blot.Hematoxylin eosin (HE) staining was performed to evaluate the pathologic change of the liver.Serum ALT levels were detected by a Roche DPPI biochemical analyzer.The level of serum IL-17 was measured by double antibody sandwich ELISA.The expressions of mRNA of IL-17R,IFN-γand IL-10 in liver were detected by RT-PCR.Results:Compared with MCMV-infected mice and isotype control,the blockade of IL-17 inhibited the expression of IL-17 protein in liver (P＜0.05).The degree of liver damage reduced obviously.The serum ALT was significantly lower [(146±15)vs (102±11)vs (37±12),P＜0.05].The level of serum IL-17 was relatively reduced[(719.76±6.06)vs (722.1±4.62) vs (707.53 ±8.58),P＜0.05].The expression of IFN-γmRNA [(0.56± 0.06)vs (0.55±0.13)vs (0.96±0.2),P＜0.05] and IL-10 mRNA[(0.55±0.073) vs (0.51 ±0.07) vs (0.903 ±0.18),P＜0.05] increased significantly,while that of IL-17R did not change apparently[(0.81±0.16)vs (0.89±0.38) vs (0.87±0.23),P＞0.05].Conclusion:The increased expression of pro-inflammatory cytokine IL-17 is involved in the pathogenesis of immune injury in cytomegalovirus hepatitis.The blockade of IL-17 is helpful to relieve the liver damage and improve the liver function.

In the present study, four new sesquiterpenoids, chimonols A-D (compounds 1-4), together with four known compounds (5-8) were isolated from the EtOAc extract of Chimonanthus praecox Link. The structures of these new compounds were elucidated on the basis of spectroscopic techniques (UV, IR, MS, and 1D and 2D NMR), and their absolute configurations were established by comparing experimental and calculated electronic circular dichroism (ECD) spectra. Compounds 1-8 were evaluated for antimicrobial activities and the minimum inhibitory concentrations (MICs) were determined by the broth microdilution method in 96-well culture plates. Compounds 1, 2, and 7 exhibited weak antibacterial effects for S. aureus (ATCC 6538), E. coli (ATCC 11775), and P. aeruginosa (ATCC 10145) with MIC values being 158-249 µg·mL. Compounds 3-7 showed activities against C. glabrata (ATCC 2001) and S. aureus (ATCC 43300) with MIC values being 128-197 µg·mL. Compounds 1-4 showed activity against S. aureus (ATCC 25923) with MIC values being 162-254 µg·mL. The present study provided a basis for future evaluation of these compounds as antibacterial agents.
Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Calycanthaceae ; chemistry ; Escherichia coli ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Sesquiterpenes ; chemistry ; isolation & purification ; pharmacology ; Staphylococcus aureus ; drug effects

To address the increasing need for detecting and validating protein biomarkers in clinical specimens, mass spectrometry (MS)-based targeted proteomic techniques, including the selected reaction monitoring (SRM), parallel reaction monitoring (PRM), and massively parallel data-independent acquisition (DIA), have been developed. For optimal performance, they require the fragment ion spectra of targeted peptides as prior knowledge. In this report, we describe a MS pipe-line and spectral resource to support targeted proteomics studies for human tissue samples. To build the spectral resource, we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on Docker. We then applied the workflow to gen-erate DPHL, a comprehensive DIA pan-human library, from 1096 data-dependent acquisition (DDA) MS raw files for 16 types of cancer samples. This extensive spectral resource was then applied to a proteomic study of 17 prostate cancer (PCa) patients. Thereafter, PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was validated. As a second application, the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma (DLBCL) patients and 18 healthy control subjects. Differentially expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker discovery. DPHL is freely accessible at https://www.iprox.org/page/project.html?id=IPX0001400000.