Intranasal immunisation with recombinant Toxoplasma gondii uridine phosphorylase confers resistance against acute toxoplasmosis in mice.

Toxoplasmosis is caused by Toxoplasma gondii, which infects all warm-blooded animals, including humans. Currently, control measures for T. gondii infection are insufficient due to the lack of effective medications or vaccines. In this paper, recombinant T. gondii uridine phosphorylase (rTgUPase) was expressed in Escherichia coli and purified via Ni2+-NTA agarose. rTgUPase was inoculated intranasally into BALB/c mice, and the induced immune responses were evaluated by mucosal and humoral antibody and cytokine assays and lymphoproliferative measurements. Moreover, the protective effect against the T. gondii RH strain infection was assessed by calculating the burdens of tachyzoites in the liver and brain and by recording the survival rate and time. Our results revealed that mice immunised with 30 µg rTgUPase produced significantly higher levels of secretory IgA (sIgA) in nasal, intestinal, vaginal and vesical washes and synthesised higher levels of total IgG, IgG1 and, in particular, IgG2a in their blood sera. rTgUPase immunisation increased the production of IFN-gamma, interleukin IL-2 and IL-4, but not IL-10 from isolated mouse spleen cells and enhanced splenocyte proliferation in vitro. rTgUPase-inoculated mice were effectively protected against infection with the T. gondii RH strain, showing considerable reduction of tachyzoite burdens in liver and brain tissues after 30 days of infection, and a 44.29% increase in survival rate during an acute challenge. The above findings show that intranasal inoculation with rTgUPase provoked mucosal, humoral and cellular immune responses and indicate that rTgUPase might serve as a promising vaccine candidate for protecting against toxoplasmosis.

Title: L'immunisation intranasale avec l'uridine phosphorylase recombinante de Toxoplasma gondii confère une résistance contre la toxoplasmose aiguë chez la souris. Abstract: La toxoplasmose est causée par Toxoplasma gondii, qui infecte tous les animaux à sang chaud, y compris les humains. Actuellement, les mesures de contrôle de l'infection à T. gondii sont insuffisantes en raison du manque de médicaments ou de vaccins efficaces. Dans cet article, l'uridine phosphorylase recombinante de T. gondii (rTgUPase) a été exprimée dans Escherichia coli et purifiée via de l'agarose Ni2+-NTA. La rTgUPase a été inoculée par voie intranasale à des souris BALB/c et les réponses immunitaires induites ont été évaluées par des dosages d'anticorps et de cytokines muqueuses et humorales et par des mesures de lymphoprolifération. De plus, l'effet protecteur contre l'infection par la souche RH de T. gondii a été évalué en calculant la charge de tachyzoïtes dans le foie et le cerveau et en enregistrant le taux et la durée de survie. Nos résultats ont révélé que les souris immunisées avec 30 µg de rTgUPase produisaient des taux significativement plus élevés d'IgA sécrétoires (sIgA) dans les lavages nasaux, intestinaux, vaginaux et vésicaux et synthétisaient des taux plus élevés d'IgG totales, d'IgG1 et, en particulier, d'IgG2a dans leur sérum sanguin. L'immunisation par la rTgUPase a augmenté la production d'IFN-gamma, d'interleukine IL-2 et IL-4, mais pas d'IL-10 à partir de cellules de rate de souris isolées et a amélioré la prolifération des splénocytes in vitro. Les souris inoculées par la rTgUPase ont été efficacement protégées contre l'infection par la souche RH de T. gondii, montrant une réduction considérable de la charge de tachyzoïtes dans les tissus hépatiques et cérébraux après 30 jours d'infection et une augmentation de 44,29 % du taux de survie lors d'une épreuve aiguë. Les résultats ci-dessus montrent que l'inoculation intranasale de rTgUPase provoque des réponses immunitaires muqueuses, humorales et cellulaires et indiquent que la rTgUPase pourrait servir de candidat vaccin prometteur pour la protection contre la toxoplasmose.

ANKRD49 promotes the invasion and metastasis of lung adenocarcinoma via a P38/ATF-2 signalling pathway.

Lung adenocarcinoma (LUAD) is the most challenging neoplasm to treat in clinical practice. Ankyrin repeat domain 49 protein (ANKRD49) is highly expressed in several carcinomas; however, its pattern of expression and role in LUAD are not known. Tissue microarrays, immunohistochemistry, χ2 test, Spearman correlation analysis, Kaplan-Meier, log-rank test, and Cox's proportional hazard model were used to analyse the clinical cases. The effect of ANKRD49 on the LUAD was investigated using CCK-8, clonal formation, would healing, transwell assays, and nude mice experiment. Expressions of ANKRD49 and its associated downstream protein molecules were verified by real-time PCR, Western blot, immunohistochemistry, and/or immunofluorescence analyses. ANKRD49 expression was highly elevated in LUAD. The survival rate and Cox's modelling analysis indicated that there may be an independent prognostic indicator for LUAD patients. We also found that ANKRD49 promoted the invasion and migration in both in in vitro and in vivo assays, through upregulating matrix metalloproteinase (MMP)-2 and MMP-9 activities via the P38/ATF-2 signalling pathway Our findings suggest that ANKRD49 is a latent biomarker for evaluating LUAD prognosis and promotes the metastasis of A549 cells via upregulation of MMP-2 and MMP-9 in a P38/ATF-2 pathway-dependent manner.

[Effects of different disturbance patterns on stand structure of infected pure Pinus massoniana plantation]. / 不同干扰模式对受害马尾松人工纯林林分结构的影响.

To explore the effects of different disturbance patterns on restoring the health of an infected stand, concentrated disturbance of not cutting trees before 10 years after infection, mode-rate disturbance of cutting infected pine trees, and strong distrubance of cutting infected pine trees, the neighboring trees and poorly growing pine trees were compared in a pure Pinus massomiana plantation infected by Bursaphelenchus xylophlius in Anji, Zhejiang, China. After 16 years, the importance values of P. massoniana in the three treatments were: concentrated disturbance > mode-rate disturbance > strong disturbance. However, the importance values of broad-leaved trees showed the opposite trend. Compared with the concentrated disturbance, the average DBH of P. massoniana in the moderate and strong disturbance treatments were 1.2 and 1.7 times higher, respectively, and those of broad-leaved species were 1.3 and 1.9 times higher, respectively. The average height of pine trees in the moderate and strong disturbance treatments increased 1.5 and 2.0 times, respectively, and those of broad-leaved species 1.2 and 1.8 times, respectively. The tree volume per hectare in moderate and strong disturbance treatments were 5.2 and 3.8 times that of concentrated disturbance treatment, respectively. In the moderate and strong disturbance treatments, the number of trees in each diameter class was greater than in the concentrated disturbance treatment. The stand diameter distribution in the multi-storied moderate and strong disturbance treatments followed an inverse J-shaped curve. The species richness and biodiversity were significantly higher in the mode-rate and strong disturbance treatments than in the concentrated disturbance treatment. The individual size inequality and structural complexity indices followed the order of moderate disturbance > strong disturbance > concentrated disturbance. Under moderate and strong disturbance treatments, the single-storied and evenly aged pure P. massoniana plantation became multi-storied and unevenly aged mixed stands. All the three disturbance patterns promoted the succession of broad-leaved trees, with the pace of succession in the order of strong disturbance > moderate disturbance > concentrated disturbance. In conclusion, moderate disturbance achieved better restoration. Thinning pure P. massoniana plantation could accelerate the succession of a mixed stand to enhance resistance against Bursaphelenchus xylophlius invasion.

[Effects of target tree tending on community structure and diversity in subtropical natural secondary shrubs]. / ç›®æ ‡æ ‘æŠšè‚²å¯¹äºšçƒ­å¸¦å¤©ç„¶æ¬¡ç”ŸçŒä¸›ç¾¤è½ç»“æž„å’Œå¤šæ ·æ€§çš„å½±å“.

The typical natural secondary shrub community was chosen in Lin'an of Zhejiang Pro-vince to discover its possibility of restoration to arbor forest with three kinds of forest management models being taken, i.e., no care as control, closed forest management and target tree tending. Over four years growth, compared with control, closed forest management significantly increased average DBH and height by 130% and 50%, respectively, while 260% and 110% for target tree tending. In target tree tending plots, larger trees had been emerging with 4.5-8.5 cm diameter class and 4.5-8.5 m height class and formed a new storey of 4 m compared with control. The species biodiversity indexes at shrub layer were significantly increased in closed management plots, and did not decrease in target tree tending plots. Closed forest management did not change the tree species composition, following its previous succession direction. However, target tree tending increased the importance value of target species with the high potential succession direction of mixed coniferous-broadleaved forest. The results revealed that the secondary shrub community with target tree tending achieved more desired goals on DBH and height growth of dominant trees and species composition improvement compared with closed management. If the secondary shrub community could be managed when the operational conditions existed, target tree tending model should be selected to accelerate the restoration of shrub toward arbor forest.