Construction of antibiotic-induced depression mice model and the function of intestinal microbiota.

Many research studies focus on intestinal microbiota-related depression induced by the usage of antibiotics, but the use of antibiotics is fairly different. To construct an effective antibiotic-induced depression mice model and explore the effect of intestinal microbiota in antibiotic-induced depression, we used several kinds of antibiotic mixtures to induce mice depression and used depression-related behavioral tests and neurobiological factors to evaluate the construction of the antibiotic-induced depression mice model. SPSS statistical software was used to analyze the above data, and the optimal model was selected according to the stability of the results and the simplicity of the modeling methods. Metagenomic analysis and fecal microbiota transplantation (FMT) of intestinal microbiota from antibiotic-induced depression mice were performed to analyze the effect of intestinal microbiota. The results showed that antibiotic mixture A (1.25 µg/mL natamycin, 5 mg/mL neomycin sulfate, and 5 mg/mL bacitracin), antibiotic mixture B (24 mg/mL bacitracin, 24 mg/mL neomycin sulfate, 9.6 mg/mL ampicillin, 4.8 mg/mL meropenem, and 1.47 mg/mL vancomycin), and antibiotic solution D (only containing 5 mg/mL neomycin sulfate) could induce depression-like behavior in mice. By using these antibiotics, the concentrations of norepinephrine (NE), 5-hydroxytryptamine (5-HT), and brain-derived neurotrophic factor (BDNF) in mice hippocampus and prefrontal cortex tissues were significantly decreased. All the above results were consistent with those of chronic unpredictable mild stress (CUMS) depression mice. The FMT results showed that fecal microbiota from antibiotic-induced depressed mice transplanted into normal mice (8 weeks-old male C57BL/6J SPF mice) also could induce depression-like behavior and cause similar changes in neurobiological factors. Metagenomic analysis showed that the community structure of microbiota in the intestinal tract of antibiotic-induced depression mice was significantly different from that in control mice, the intestinal microbiota species diversity in antibiotic-induced depression mice was lower, the lipoic acid metabolism pathway was significantly activated, and the abundance of functional gene lipA was explicitly increased. Quantitative real-time PCR (qPCR) further verified the abundance of enriched bacteria in the intestinal microbiota of antibiotic-induced depression mice. In summary, the specific antibiotic mixtures can induce depression by causing changes in intestinal microbiota in mice. Antibiotic-induced depressed mice show differences in intestinal microbiota abundance, high enrichment of the unique metabolic pathway, and the functional gene.

Antibiotics-induced depression in mice via the microbiota-gut-brain axis.

BACKGROUND: Intestinal dysbacteriosis is associated with depression. This study aimed to establish an antibiotics-induced depression mouse model and explore the mechanism of antibiotic-induced depression. METHODS: C57BL/6 J mice were treated with antibiotics to prepare the antibiotic-induced depression mouse model. Behavioral tests and depression-related bio-markers were examined. To understand the abundance of different bacteria in intestinal flora and screen out the predominant bacterial species, metagenomic analysis of feces was carried out. Finally, we detected the expression of NF-κB-p65 and p-NF-κB-p65 in PFC and the hippocampus using Western blot. RESULTS: Mixtures A and B caused depression-like behavior in mice. Norepinephrine, 5-hydroxytryptamine, and brain-derived neurotrophic factor in hippocampus and PFC of antibiotic-induced depression mice significantly decreased. The serum adrenocorticotropic hormone and corticosterone concentrations increased. The abundance values of Bacteroides thetaiotaomicron, Klebsiella oxytoca, and Klebsiella aerogenes in antibiotic-induced depression mice significantly increased, and the characteristic KO genes and metabolic pathways in antibiotic-induced depression mice were significantly different with in CUMS depression mice (the positive control) and normal mice. The relative levels of p-NF-κB-p65 in antibiotics-induced depression mice were significantly higher than in normal mice. LIMITATIONS: How dysbacteriosis induces inflammation in the central nervous system is unclear. CONCLUSIONS: Specific antibiotic mixture can cause depression-like behavior and changes of depression-related bio-markers in mice. The antibiotic-induced depression mice display changes in the species and metabolism of intestinal bacterial flora. The activation of NF-κB inflammatory signaling pathway in the central nervous system may act as one of the mechanisms in the development of antibiotic-induced depression.

Identification of lncRNA expression profiles and analysis of ceRNA in the hippocampus of perinatal glyphosate-exposed mice.

OBJECTIVE: In order to understand the role of long noncoding RNAs (lncRNAs) played in the mechanisms of glyphosate neurotoxicity in neuronal development. METHODS: Perinatal glyphosate exposure (PGE) mouse model was constructed, and a lncRNA microarray was used to study the lncRNA expression changes in the hippocampus tissue of perinatal glyphosate exposure mice. Then we used GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) databases to analyze the function of the differentially expressed mRNAs and lncRNAs. RESULTS: LncRNA microarray analysis revealed that 1759 lncRNAs and 759 mRNAs were differentially expressed in the perinatal glyphosate exposure (PGE) mice group (G group) compared with the normal control mice group (C group). The functions of the DEmRNAs are involved in the cellular response to hormone stimulus. The ceRNA analysis showed that some interaction networks existed, including (ENSMUST00000137546, ENSMUST00000160950)/(miR-34a-3p, miR-130a-3p)/(Il12b, Irf1). Further analysis of the target mRNAs of miRNAs indicated that the possible functions involved the neuroactive ligand-receptor interaction and calcium signaling pathway, which are involved in perinatal glyphosate exposure-induced neurotoxicity. CONCLUSION: The aberrant expression of lncRNAs is related to the perinatal glyphosate-exposed neurotoxicity. These lncRNAs affect the target gene expression level, might by regulating neuroactive ligand-receptor interactions. The (ENSMUST00000137546, ENSMUST00000160950)/ (miRNA-34a-5p, miR-130a-3p) / mRNAs (e.g., Il12b, Irf1) interaction network may functions in perinatal glyphosate exposure-induced neurotoxicity.

Human immunodeficiency virus infection and mortality risk among lung cancer patients: A systematic review and meta-analysis.

BACKGROUND: Previous studies have suggested that patients with human immunodeficiency virus (HIV) infection are at higher risk of lung cancer, but the impact of HIV infection on the risk of mortality among lung cancer patients is still unclear. We conducted a systematic review and meta-analysis to clarify the association between HIV infection and mortality risk among lung cancer patients. METHODS: PubMed and Embase databases were searched to identify studies assessing the association between HIV infection and mortality risk among lung cancer patients. Only studies reporting adjusted relative risk (RR) of mortality among lung cancer patients with HIV infection were included. Meta-analysis of random-effect model was utilized to calculate the pooled RR with 95% confidence interval (CI). RESULTS: Twelve cohort studies were finally included. Compared with lung cancer patients without HIV infection, the pooled RR of mortality among lung cancer patients with HIV infection was 1.48 (95% CI, 1.22-1.78, P < .001; I = 88.6%). After excluding 2 studies with low quality, HIV infection was still significantly associated with an elevated risk of mortality among lung cancer patients (RR = 1.51, 95% CI, 1.25-1.82, P < .001; I = 89.8%). Sensitivity analysis showed that the statistical significance of the pooled RR was not changed by excluding any one study. CONCLUSION: The outcomes from the meta-analysis provide strong evidence for the elevated risk of mortality among lung cancer patients with HIV infection, and HIV infection is an important prognostic factor in lung cancer patients.

Effect of camptothecin on inducible nitric oxide synthase expression in the colon cancer SW480 cell line.

As a topoisomerase I inhibitor, camptothecin (CPT) is regarded as an effective antitumor agent. In an attempt to search for its novel anticancer mechanism, the present study evaluated the effects of CPT on inducible nitric oxide synthase (iNOS) in the human colon cancer SW480 cell line when stimulated with lipopolysaccharide (LPS) and interleukin (IL)-1ß. The data indicated that CPT significantly decreased NO production. Consistent with these observations, the protein and mRNA expression levels of iNOS were inhibited by CPT in a dose-dependent manner. Thus, the inhibitory effects of CPT on LPS/IL-1ß-stimulated NO production were likely mediated via the inhibition of iNOS gene transcription. From these results, we propose that the inhibition of NO biosynthesis by CPT may partially underlie the efficacy of this antitumor agent.

Perifosine inhibits S6K1-Gli1 signaling and enhances gemcitabine-induced anti-pancreatic cancer efficiency.

PURPOSE: The pancreatic cancer has extremely low overall 5-year survival, and gemcitabine is the only approved single agent for pancreatic cancer treatment. METHODS: In the present study, we investigated the potential effect of perifosine, a novel Akt inhibitor on gemcitabine-induced anti-pancreatic cancer effect both in vivo and in vitro. RESULTS: We showed that sub-cytotoxic low concentration of perifosine dramatically enhanced gemcitabine-induced cytotoxicity in cultured pancreatic cancer cells. Perifosine inhibited Akt-mammalian target of rapamycin and Erk-mitogen-activated protein kinase activation in pancreatic cancer cells. Meanwhile, perifosine suppressed the hedgehog signaling, as it inhibited glioma-associated oncogenes (Gli) 1 activation and decreased its target protein patched 1 (PTCH1) expression. Our data demonstrated that perifosine blocked p70S6K1 (S6K1) activation, thus disrupting S6K1-Gli1 association and subsequent Gli1 activation. The reduction of S6K1 or Gli1 expression by target siRNAs inhibited PTCH1 expression and enhanced gemcitabine-induced cytotoxicity in pancreatic cancer cells. Significantly, perifosine dramatically enhanced gemcitabine-mediated antitumor effect in a PANC-1 xenograft severe combined immunodeficiency mice model. CONCLUSIONS: In summary, we conclude that perifosine sensitizes gemcitabine-mediated anti-pancreatic cancer efficiency through regulating multiple signaling pathways.

Huanglian-Jie-Du-Tang extract protects against chronic brain injury after focal cerebral ischemia via hypoxia-inducible-factor-1α-regulated vascular endothelial growth factor signaling in mice.

Huanglian-Jie-Du-Tang (HJDT) is a traditional Chinese herbal formula which is widely used clinically. In this study, we investigated the effects of an aqueous (HJDTaq) and an ethanolic (HJDTet) extract of HJDT on chronic brain injury after focal cerebral ischemia in mice. The ischemia was induced by occlusion of the right middle cerebral artery for 30 min. HJDTaq (4 g/kg) and HJDTet (200, 400, 800 mg/kg) were orally administered for 21 d from day 7 before ischemia to day 14 after ischemia. The survival rate decreased to less than 50% at 35 d after ischemia. HJDTet at 400 mg/kg increased the survival rate. HJDTaq (4 g/kg) and HJDTet (400, 800 mg/kg) significantly attenuated the neurological dysfunction, brain atrophy and infarct volume after ischemia. There were few cells positive for CD31, hypoxia-inducible-factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and Flk-1 in the sham control. After ischemia, the number increased. HJDTaq (4 g/kg) and HJDTet (400 or 800 mg/kg) further increased the numbers of CD31, HIF-1α, VEGF and Flk-1-positive cells in the ischemic hemisphere. We conclude that HJDTaq and HJDTet have neuroprotective effects on chronic brain injury after focal cerebral ischemia and lead to accelerated angiogenesis by HIF-1α-regulated VEGF signaling.

NS398 induces apoptosis in non-small cell lung cancer cells.

PURPOSE: Non-steroidal anti-inflammatory drugs (NSAIDs) can induce the apoptosis of many tumor cells and inhibit their growth. NS398 is an NSAID that inhibits COX-2 expression and induces tumor apoptosis via other pathways. The current study aims to observe the effects of NS398 on A549 cell apoptosis and investigate the apoptosis mechanism. METHODS: The A549 cells were treated with different NS398 concentrations. The growth inhibition of A549 cell was analyzed via a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and morphologic alterations were observed to detect apoptosis. The expression of survivin and caspase-3 mRNA was quantified via reverse transcriptase polymerase chain reaction, and the expression of caspase-3 and p-AKT protein was detected via western blot analysis. RESULTS: The MTT results show that NS398 inhibits A549 cell growth. The inhibition rate of NS398 (400 µmol/L) on A549 cells is up to 66.95% after 48 h of treatment. Simultaneously, the morphology experiment revealed significant apoptotic characteristics in A549 cells, such as green nuclear plasmid and different degrees of nuclear fragmentation. The expression of survivin mRNA was significantly reduced (P < 0.05, P < 0.01, and P < 0.001) and that of caspase-3 mRNA was significantly increased (P < 0.05 and P < 0.001) in the group treated with NS398 for 24 h in a dose-dependent manner. On the other hand, survivin and p-AKT were expressed at low levels (P < 0.01 and P < 0.001) and caspase-3 was increased significantly (P < 0.05 and P < 0.001) in the group treated with NS398 for 48 h in a dose-dependent manner. CONCLUSION: The current study proves that NS398 induces apoptosis in A549 cells, thereby inhibiting tumor growth. This function of NS398 may be related to the inhibition of AKT phosphorylation and survivin protein downregulation.

[Comparison of behavioral effects of psychoactive drugs between two strains of mice].

OBJECTIVE: To compare the behavioral effects of psychoactive drugs between two strains of mice. METHODS: The Kunming (KM) and ICR mice were injected intraperitoneally with caffeine (3, 10, 30, 100 mg/kg), ephedrine (3, 10, 30, 100 mg/kg), diazepam (1, 3,1 0 mg/kg) and chloral hydrate (10, 30, 100 mg/kg), respectively. Ten min after injection, the locomotor activity in the open field was recorded for 2 h. The total distance, the distance ratio to total distance and the time in central region were analyzed for each drugs. Thirty min after injection, the latent time in the passive avoidance test was measured in a shuttle box. RESULTS: Caffeine and diazepam prolonged the latent time, and ephedrine and chloral hydrate decreased the latent time, but there were no differences between the two strains. The two strains of mice exhibited significant differences in the total distance after injection of ephedrine 10 mg/kg, diazepam 3 mg/kg and chloral hydrate 100 mg/kg. Compared to KM mice, ICR mice exhibited an increase in the distance ratio and the time in central region after injection of ephedrine 10-100 mg/kg, but a decrease after diazepam 3-10 mg/kg. CONCLUSION: KM and ICR mice show no differences in latent time, but significant differences in the total distance, the distance ratio and the time in central region in the locomotor activity. Therefore, selection of mouse strains is important in the study of psychoactive drugs.

[Recombinant expression of the fusion antigen based on Treponema pallidum TpN17 and TpN47 epitope peptides and establishment and application of the associated ELISA].

Using recombinant TpNs proteins of Treponema pallidum as antigens, ELISAs are proved to be of higher sensitivity and specificity. However, they can be further increased by using multiple TpNs antigens. According to the epitope analysis, we firstly used linking primers PCRs to obtain an artificial fusion gene segment tpE17-47 containing epitopes of both TpN17 and TpN47. Subsequently, we conducted the prokaryotic expression systems of entire tpN17 and tpN47 genes and tpE17-47 fusion gene. SDS-PAGE analysis and BioRad Gel Image Analysis System showed that the recombinant proteins rTpN17, rTpN47 and rTpE17-47 expressed stably, with 36%, 20% and 28% yields of total bacterial protein, respectively. After purified by Ni-NTA affinity chromatography, all the three recombinant proteins could be recognized by T. pallidum antibody positive sera from syphilis patients. The positive rate of rTpE17-47-ELISA for detecting serum specimens in clinically 630 cases with syphilis was 98.6%. This rate was slightly higher than that by Treponema pallidum particle agglutination (TPPA) (97.9%) (P > 0.05), but significantly higher than those by rTpN17-ELISA (83.8%), rTpN47-ELISA (83.3%) and rapid plasma reagin (RPR) (72.1%) (P < 0.01). Furthermore, both ELISAs and TPPA for detecting the serum specimens in 25 cases with SLE, 36 cases with RA and 250 healthy cases were all negative. RPR showed positive in 1 case with SLE, 2 cases with RA and 2 healthy cases. This could be a novel serological screening or diagnostic method of syphilis with advantages of quickness, convenience, safety, sensitivity and specificity.

[Relationship of angiopoietins expression with microvessel density (MVD) in human colorectal tumors].

OBJECTIVE: To investigate the expression of angiopoietin (Ang)-1 and Ang-2 in colorectal tumors and its relations to microvessel density (MVD) in tumor tissue. METHODS: Ang-1, Ang-2 and factor VIII-related antigen were stained immunohistochemically in 91 cases of primary colorectal adenocarcinoma, 20 cases of colorectal adenoma and 24 cases of normal colorectal mucosal tissue, and MVD was also assayed in above tissue specimens. RESULT: (1) A significantly higher Ang-1 (7.07+/-2.00) was observed in normal tissue compared with 1.75 +/-1.98 in the adenoma and 1.40 +/- 1.22 in the adenocarcinoma (P<0.01). (2) Ang-2 protein positive rate in adenocarcinoma was significantly higher than that in normal tissue and adenoma (P<0.01). The expression of Ang-2 in adenocarcinoma was closely associated with poor differentiation and vessel invasion. (3) There were significant correlations between Ang-1 and Ang-2 (r=-0.338, P<0.01), Ang-1 and MVD (r=-0.388, P<0.01), Ang-2 and MVD (r=0.594, P<0.01) in the 135 cases. CONCLUSION: The overexpression of Ang-2 may play an important role in angiogenesis of colorectal adenocarcinoma. It can be regarded as an index for malignancy and prognosis in colorectal adenocarcinoma.