Excretory/secretory proteins of adult Toxocara canis induce changes in the expression of proteins involved in the NOD1-RIP2-NF-κB pathway and modulate cytokine production in mouse macrophages.

Dog roundworm (Toxocara canis) is the major causative agent of toxocarosis, a parasitic disease of both veterinary and medical importance. Knowledge gaps in fundamental and applied aspects hinder the control of this important zoonotic disease. To have a better understanding of Toxocara infection and host immune responses, mouse macrophages were exposed to excretory/secretory (ES) proteins released by adult worms of T. canis in vitro. The messenger RNA transcription and protein expression of nucleotide-binding oligomerization domain-containing protein 1 (NOD1), receptor interacting protein 2 (RIP2) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in macrophages were analysed using quantitative real-time PCR (qRT-PCR) and Western blot. The levels of tumour necrosis factor alpha (TNF-ɑ), interleukin-1 beta (IL-1ß) and IL-6 released by the stimulated macrophages were analysed using enzyme-linked immunosorbent assay. It was found that 20 µg/mL ES proteins of adult T. canis induced the expression of NOD1, RIP2 and NF-κB in mouse macrophages at both transcriptional and translational levels after 9 h of incubation in vitro. Incubation with 20 µg/mL ES proteins also modulated the production of pro-inflammatory cytokines TNF-ɑ, IL-1ß and IL-6 by the macrophages. Taken together, ES proteins of adult T. canis appeared to be able to affect the macrophage NOD1-RIP2-NF-κB signalling pathway, which might play a role in regulating the production of proinflammatory cytokines. Further investigation of these aspects should lead to a better understanding of immune recognition of and modulation by Toxocara canis in host animals.

[The clinical study of modified transseptal suturing after septoplasty].

Objective:To study whether modified transseptal suturing could be an alternative to packing after septoplasty. Method:We conducted a prospective randomized controlled trial. The study involved 40 patients who underwent septoplasty. Patients were randomly divided into two groups, one with anterior nasal packing and the other with modified transseptal suturing. Patients were asked to record their subjective discomfort levels on a visual analogue scale（VAS） during 48 h postoperatively. Both groups were compared for VAS, postoperative complications 2 weeks after septoplasty through nasal endoscope, the datum of rhinomanometry before surgery and 2 weeks after surgery. Result:The differences of mean postoperative VAS score between two groups were statistically significant（P<0.05）. The patients of nasal packing group experienced more discomforts than the other group. No statistically significant between two groups in terms of the datum of rhinomanometry before surgery and 2 weeks after surgery（P>0.05）. There were no bleeding, septal hematoma, nasal septum perforation and local infection in all patients 2 weeks after septoplasty. During follow-up 1 case was found with postoperative nasal adhesion, nasal vestibular swelling and fester were seen in two patients and nasal peculiar smell was found in 1 case in nasal packing group. We can find that there were a lot of blood crust in silicone ventilation tube about in 14 patients in packing group. However, the suturing group could see none of the above questions. Conclusion:Modified transseptal suturing is more simple and convenient than nasal packing. In a word, the use of modified transseptal suturing could be an alternative topacking after septoplasty.

Complete genome sequence of an M1 strain of Streptococcus pyogenes.

The 1,852,442-bp sequence of an M1 strain of Streptococcus pyogenes, a Gram-positive pathogen, has been determined and contains 1,752 predicted protein-encoding genes. Approximately one-third of these genes have no identifiable function, with the remainder falling into previously characterized categories of known microbial function. Consistent with the observation that S. pyogenes is responsible for a wider variety of human disease than any other bacterial species, more than 40 putative virulence-associated genes have been identified. Additional genes have been identified that encode proteins likely associated with microbial "molecular mimicry" of host characteristics and involved in rheumatic fever or acute glomerulonephritis. The complete or partial sequence of four different bacteriophage genomes is also present, with each containing genes for one or more previously undiscovered superantigen-like proteins. These prophage-associated genes encode at least six potential virulence factors, emphasizing the importance of bacteriophages in horizontal gene transfer and a possible mechanism for generating new strains with increased pathogenic potential.

Evidence of gamma-aminobutyric acidergic control over the catecholaminergic projection from the medulla oblongata to the central nucleus of the amygdala.

It is known that the nucleus of the solitary tract (NTS) and the ventrolateral medulla (VLM) project to the central nucleus of the amygdala (Ce), conveying visceral information. Conversely, the Ce sends projections to the NTS and the VLM. To understand better the role of catecholamine and gamma-aminobutyric acid (GABA) in these reciprocal connections, experiments were performed by combining lectin-conjugated horseradish peroxidase (WGA-HRP) anterograde and retrograde transport with preembedding immunocytochemistry to detect tyrosine hydroxylase (TH), and postembedding immunocytochemistry to detect GABA. The light microscopic study suggested that the majority of neurons in the NTS and the VLM projecting to the Ce were TH immunoreactive (TH-IR). Most of them were located at the level of the obex. Under the electron microscope, the GABAergic and non-GABAergic terminals were found to form synaptic contacts with the TH-(IR) or Ce-projecting or TH-IR/Ce-projecting double-labelled neurons of the NTS and VLM. The GABAergic terminals mostly formed symmetrical synaptic contacts with the postsynaptic structure in which perikarya (14-19%), dendrites (79-84%), and spines (2%) were observed. Approximately 94% of the axon terminals in the NTS and 90% of those in the VLM arising from the Ce were GABAergic and appeared not to form synaptic contacts with the TH-IR or Ce-projecting neurons in these regions. The present results demonstrated that the catecholaminergic neurons of the NTS and VLM projecting to the Ce receive an extensive GABAergic innervation and that the amygdala projection to the medulla is mostly GABAergic.

GABAergic synapses upon neurons expressing substance P receptors in the nucleus of the solitary tract: an immunocytochemical electron microscope study in the rat.

Morphological substrates for interactions between gamma-aminobutyric acid (GABA) and substance P upon neurons expressing substance P receptor (SPR) in the nucleus of the solitary tract (NST) were investigated by immunocytochemical electron microscopy. In the NST of the rat, many GABA-like immunoreactive axon terminals were in symmetric synaptic contacts with dendritic profiles; they were observed on nearly a half of the SPR-like immunoreactive dendritic profiles in the medial part of the caudal half of the NST.

An indirect projection from the nucleus of the solitary tract to the central nucleus of the amygdala via the parabrachial nucleus in the rat: a light and electron microscopic study.

The morphological basis of how visceral information from the nucleus of the solitary tract (NTS) is relayed from the parabrachial nucleus (PBN) to the central nucleus of the amygdala (Ce) was studied at the light and electron microscopic levels using the anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHA-L), kainic acid degeneration, and retrograde tracing with horseradish peroxidase (HRP). After injection of PHA-L into the caudal NTS, anterogradely labeled fibers and terminals were predominantly distributed in the external lateral (el) and central lateral (cl) subnuclei of the PBN. After injection of HRP into the Ce, retrogradely labeled neurons in PBN were mainly distributed in the same areas. In double-labeling experiments, there was a clear overlap between neuronal elements labeled with HRP and PHA-L in the el and cl. At the electron microscopic level, the PHA-L-labeled axon terminals from the NTS mainly contained spherical agranular synaptic vesicles and formed asymmetric contacts with the postsynaptic dendrites or dendritic spines in PBN. After the lesioning agent kainic acid was injected into the NTS and HRP deposited in the Ce, it was found the afferent fibers from the NTS made direct synaptic contact with the lateral PBN neurons which in turn projected to Ce. Such evidence adds to our growing knowledge of regulation of visceral function in central nervous system and would be likely helpful for understanding the important roles of the NTS, PBN and Ce in the central control of cardiovascular, respiratory and gastrointestinal functions.

Projection from the ventrolateral medullary neurons containing tyrosine hydroxylase to the central amygdaloid nucleus in the rat.

By using a double-labeling produce of retrograde horseradish peroxidase (HRP) tracing and the immunocytochemical localization of tyrosine hydroxylase (TH), the present study ascertained that the axonal fibers of catecholaminergic neurons in the caudal ventrolateral medulla projected to the central amygdaloid nucleus in the rat. The majority of double-labeled cells were observed primarily around the level of the obex. 92% of HRP retrogradely labeled cells contained TH-like immunoreactive (TH-IR), but only 31% of TH-IR cells was labeled with HRP.

Serotonin-, substance P- or leucine-enkephalin-containing neurons in the midbrain periaqueductal gray and nucleus raphe dorsalis send projection fibers to the central amygdaloid nucleus in the rat.

By a double-labeling method combining the retrograde tracing of horseradish peroxidase and the immunocytochemical technique, serotonin-, substance P- or leucine-enkephalin-like immunoreactive neurons in the midbrain periaqueductal gray (PAG) and the nucleus raphe dorsalis (DR) of the rat were found to send projection fibers to the central amygdaloid nucleus bilaterally with an ipsilateral dominance. These PAG neurons were chiefly distributed in the ventrolateral PAG subdivision and the ventral parts of medial PAG subdivision at the middle and caudal levels of PAG.