RESUMEN
BACKGROUND: The effects of a solitary neonatal exposure to anesthesia plus surgery (anesthesia/surgery) on cognitive function and the underlying mechanism in developing brains remains largely undetermined. We, therefore, set out to investigate the impact of single exposure to anesthesia/surgery in neonatal mice. METHODS: Six-day-old male and female mice received abdominal surgery under 3% sevoflurane plus 50% oxygen for 2 h. The new object recognition (NOR) and Morris water maze (MWM) were used to evaluate cognitive function in young adult mice. Western blot, ELISA and RT-PCR were used to measure levels of NR2B and IL-6 in medial prefrontal cortex and IL-6 in blood of the mice. We employed NR2B siRNA and IL-6 antibody in the interaction studies. RESULTS: The anesthesia/surgery decreased the ratio of novel time to novel plus familiar time in NOR and the number of platform crossings, but not escape latency, in MWM compared to sham condition. The mice in anesthesia/surgery group had increased NR2B expression in medial prefrontal cortex, and IL-6 amounts in blood and medial prefrontal cortex. Local injection of NR2B siRNA in medial prefrontal cortex alleviated the anesthesia/surgery-induced cognitive impairment. IL-6 antibody mitigated the anesthesia/surgery-induced upregulation of NR2B and cognitive impairment in young adult mice. CONCLUSIONS: These results suggest that a single neonatal exposure to anesthesia/surgery causes impairment of memory, but not learning, in young adult mice through IL-6-regulated increases in NR2B concentrations in medial prefrontal cortex, highlighting the need for further research on the underlying mechanisms of anesthesia/surgery's impact on cognitive function in developing brains.
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Anestesia , Anestésicos por Inhalación , Disfunción Cognitiva , Animales , Ratones , Masculino , Femenino , Animales Recién Nacidos , Anestésicos por Inhalación/toxicidad , Interleucina-6/genética , Anestesia/efectos adversos , ARN Interferente PequeñoRESUMEN
High-altitude polycythemia (HAPC) is characterized by excessive proliferation of erythrocytes, resulting from the hypobaric hypoxia condition in high altitude. The genetic variants and molecular mechanisms of HAPC remain unclear in highlanders. We recruited 141 Tibetan dwellers, including 70 HAPC patients and 71 healthy controls, to detect the possible genetic variants associated with the disease; and performed targeted sequencing on 529 genes associated with the oxygen metabolism and erythrocyte regulation, utilized unconditional logistic regression analysis and GO (gene ontology) analysis to investigate the genetic variations of HAPC. We identified 12 single nucleotide variants, harbored in 12 genes, associated with the risk of HAPC (4.7 ≤ odd ratios ≤ 13.6; 7.6E - 08 ≤ p-value ≤ 1E - 04). The pathway enrichment study of these genes indicated the three pathways, the PI3K-AKT pathway, JAK-STAT pathway, and HIF-1 pathway, are essential, which p-values as 3.70E - 08, 1.28 E - 07, and 3.98 E - 06, respectively. We are hopeful that our results will provide a reference for the etiology research of HAPC. However, additional genetic risk factors and functional investigations are necessary to confirm our results further. Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-021-01474-1.
RESUMEN
The relapse of fear memory remains a clinical challenge in treatment of fear-related disorders. Here we tested the effects and underlying mechanisms of probiotics treatment after fear conditioning on fear extinction. We found that fear conditioning induced synapse loss, microglial activation, and synaptic phagocytosis of activated microglial cells in hippocampal dentate gyrus of mice. And probiotics treatment (1 capsule/day/mice) after fear conditioning for 27 days inhibited these changes, promoted fear extinction, and inhibited the recovery of fear memory even 7 days after extinction. 16S rRNA gene sequencing demonstrated that probiotics supplement after fear conditioning partially normalized fear conditioning-induced dysbiosis of gut microbiota. In addition, we also found that repopulation of microglial cells in fear conditioning mice via PLX3397 treatment promoted long-term extinction of fear memory. Probiotics treatment after fear conditioning inhibited microglial activation and had similar therapeutic effects as the microglial cell repopulation induced by PLX3397 treatment. These data showed that (1) probiotics treatment after fear conditioning might promote long-term fear extinction which could be associated with the mitigation of synaptic pruning of activated microglial cells; (2) probiotics may be applicable as therapeutic strategy to inhibit microglial activation and treat fear-related disorders.
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Extinción Psicológica/efectos de los fármacos , Miedo/efectos de los fármacos , Microglía/efectos de los fármacos , Probióticos/farmacología , Animales , Giro Dentado/efectos de los fármacos , Homólogo 4 de la Proteína Discs Large/metabolismo , Microbioma Gastrointestinal/efectos de los fármacos , Hipocampo/efectos de los fármacos , Masculino , Memoria/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Plasticidad Neuronal/efectos de los fármacos , Recurrencia , Sinapsis/efectos de los fármacos , Sinaptofisina/metabolismoRESUMEN
We have studied the fluorescence of Mg-Al-Eu ternary layered hydroxides (TLH) quenched by tryptophan (Trp). IR spectroscopy was used to evaluate the change of Trp structure which was caused by TLH. XRD and TG-DTA results further suggested a structural change of Trp after being reacted with TLH. XPS characterization confirmed a strong chemical reaction between Trp and TLH. These studies may present more direct evidence to explain the interrelation between the structural change of Trp and the fluorescent quenching of TLH.
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Aluminio/química , Europio/química , Fluorescencia , Hidróxidos/química , Magnesio/química , Triptófano/análisis , Espectrofotometría , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Termogravimetría , Rayos XRESUMEN
We reported for the first time the fluorescence sensitivity of a Mg-Al-Eu ternary layered double hydroxide (LDH) to phenylalanine (Phe). As shown in fluorescent spectra, the red emissions resulted from (5)D(0)-(7)F(J) transition (J=1, 2) of Eu(3+) were quenched by Phe, and a strong peak around at 492 nm emerged. A strong interaction between the Mg-Al-Eu LDH and Phe was manifested by markedly different chemical shift positions of the Mg 2p, Al 2p, Eu 4d, O 1 s, and C 1 s peaks in the XPS spectra from those of the LDHs. These results showed that the charges transfer from Phe to the trivalent Eu ions may be taken place.
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Aluminio/química , Europio/química , Fluorescencia , Hidróxidos/química , Magnesio/química , Fenilalanina/análisis , Espectrometría de Fluorescencia , Espectrofotometría , Rayos XRESUMEN
1071-bp fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3' and 5' ends of the incomplete expression sequence tag (EST) of succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) were amplified by the anchored PCR with 2 pairs of primers designed according to the EST of SjSDISP and the sequence of multiclone sites of the library vector. Sequence analysis indicated that the fragment was a full-length cDNA with a complete open reading frame (ORF), encoding 278 amino acid residues. The fragment was cloned into prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE and Western-blot analyses showed that the recombinant protein was about 32 kD and could be recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Compared with the FCA controls, mice vaccinated with rSjSDISP (test) or rSjGST (positive control) all revealed high levels of specific antibody and significant reduction in worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs. These results suggest that SjSDISP may be a novel and partially protective vaccine candidate against schistosomiasis. In contrast to the worm burden reduction rate, the higher degree of egg reduction rate in the test group also suggested that SjSDISP vaccine may primarily play a role in anti-embryonation or anti-fecundity immunity.
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Schistosoma japonicum/enzimología , Schistosoma japonicum/genética , Succinato Deshidrogenasa/genética , Animales , Clonación Molecular , Cartilla de ADN , ADN Complementario/genética , ADN de Helmintos/genética , ADN de Helmintos/aislamiento & purificación , Femenino , Biblioteca de Genes , Proteínas del Helminto/genética , Humanos , Proteínas Hierro-Azufre/genética , Ratones , Ratones Endogámicos BALB C , PlásmidosRESUMEN
To observe the in vitro expression of DNA vaccine pcDNA3-Sj22.7 and its immunological effect in mice, the recombinant plasmid pcDNA3-Sj22.7 was used to transfect HeLa cells with liposome-mediated method and the expression of Sj22.7 mRNA and protein was examined using reverse transcription-polymerase chain reaction, sodium dodecylsulfate-polyacrylamide gel electrophoresis and Western blot. Then, the ability of pcDNA3-Sj22.7 to protect against Schistosoma japonicum challenge infections was analyzed according to worm reduction rate and egg reduction rate after vaccination of mice. The serum levels of specific IgG antibody and T lymphocyte proliferation response were also determined. After the challenge infection, Sj22.7-driven interferon (IFN)-gamma and interleukin (IL)-4 was also quantified. Results showed that pcDNA3-Sj22.7 could express Sj22.7 mRNA and protein in vitro. Immunization resulted in a worm reduction rate of 29.70%, egg reduction rate of 47.25% (liver) and 51.73% (intestine), and egg reduction rate of 25.90% (eggs per female), suggesting induction of significant anti-fecundity in the pcDNA3-Sj22.7 group. Enzyme-linked immunosorbent assay and Western blot analysis indicated that immunized mice generated specific IgG against Sj22.7. T lymphocytes from mice immunized with pcDNA3-Sj22.7 showed a significant proliferation response to rSj22.7. The culture of spleen cells showed that secretion of IFN-gamma increased but IL-4 decreased. The results indicate hat DNA vaccination by pcDNA3-Sj22.7 is sufficient to elicit significant levels of protective immunity against S. japonicum infection. The DNA vaccine could induce significant cellular and humoral immune response, and display predominant T helper cell type 1 type immune responses, which contribute to the protective immunity against challenge infection in mice.
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Antígenos Helmínticos/genética , Schistosoma japonicum/inmunología , Esquistosomiasis Japónica/prevención & control , Vacunación , Vacunas de ADN/inmunología , Animales , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/inmunología , Granuloma/patología , Interferón gamma/biosíntesis , Interleucina-4/biosíntesis , Activación de Linfocitos , Esquistosomiasis Japónica/patología , Linfocitos T/inmunologíaRESUMEN
OBJECTIVE: To clone the full-length gene encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) Chinese strain and express it in Escherichia coli. METHODS: According to the published incomplete EST (BU804141) of SjSDISP and the sequence of multiclone sites of lambda gt11 vector, 2 pairs of primers were designed and synthesized. Then the 3' and 5'ends of the EST of the SjSDISP from adult Schistosoma japonicum cDNA library were amplified by anchored PCR. After sequencing, a full-length cDNA sequence of the SjSDISP was obtained, and then it was cloned into prokaryotic expression vector pGEX-4T-1. Identified by agarosed gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for the expression under the temperature-dependent condition and Western blot analysis. RESULTS: A 1,071 bp sequence was obtained. Sequence analysis showed that the fragment contained a complete open reading frame (ORF), encoding 278 amino acid residues. This target fragment was cloned into the prokaryotic expression vector pGEX-4T-1, and expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of the expressed fusion recombinant product was 56 kD. Western blot showed that the recombinant protein was recognized by polyclonal rabbit antiserum immunized with Schistosoma japonicum adult worm antigen. CONCLUSION: Cloning of the full-length gene encoding SjSDISP and its bacterial expression were successfully done.