Archaea is more important than bacteria in driving soil stoichiometry in phosphorus deficient habitats.

Phosphorus deficiency is a critical limit on the cycling of carbon (C), nitrogen (N) and phosphorus (P) in forest ecosystems. Despite the pivotal roles of microbes in driving the biogeochemical cycling of C/N/P, our knowledge on the relationships of soil bacteria and archaea to P deficiency in forest ecosystems remains scarce. Here, we studied 110 acidic soils (average pH 4.5) collected across 700-km subtropical forests with a gradient of available phosphorus (AP) ranging from 0.21 to 17.6 mg/kg. We analyzed the soil C/N/P stoichiometry and studied soil bacterial and archaeal diversity/abundance via high throughput sequencing and qPCR approaches. Our results show that soil P decoupled with N or C when below 3 mg/kg but coupled with C and N when above 3 mg/kg. Archaeal diversity and abundance were significantly higher in low AP (< 3 mg/kg) soils than in high AP (>3 mg/kg) soils, while bacterial were less changed. Compared with bacteria, archaea are more strongly related with soil stoichiometry (C:N, C:P, N:P), especially when AP was less than 3 mg/kg. Taxonomic and functional composition analysis further confirmed that archaeal rather than bacterial taxonomic composition was significantly related with functional composition of microbial communities. Taken together, our results show that archaea are more important than bacteria in driving soil stoichiometry in phosphorus deficient habitats and suggest a niche differentiation of soil bacteria and archaea in regulating the soil C/N/P cycling in subtropical forests.

[A calcium-sensing receptor polymorphism at E942K promotes the proliferation of gastric cancer cells via Ca2+ and ERK1/2 pathways].

The study was designed to investigate the effects and mechanism of a calcium-sensing receptor (CaSR) polymorphism at E942K on the proliferation of gastric cancer cells. Single nucleotide polymorphisms (SNPs) were detected between gastric cancers group and normal controls group by DNA sequence analysis. The cell model was constructed by transfection of E942K mutant plasmid and wild-type (WT) plasmid into SGC-7901 and HEK-293 cells. The effect of E942K mutation on cell proliferation ability was detected by CCK8 and cell clone formation experiments. The effect of E942K mutation on calcium signaling was detected by calcium imaging. Western blot experiments were used to detect changes in phosphorylation levels of key proteins ERK1/2 and ß-catenin in downstream signaling pathways after E942K mutation. The results showed that the mutation rate of E942K in gastric cancer group was significantly higher than that in normal control group (P < 0.05). CCK8 and cell clone formation experiments showed that E942K mutation significantly improved the proliferation ability of SGC-7901 gastric cancer cells and HEK-293 cells. E942K mutation enhanced calcium signaling in SGC-7901 and HEK-293 cells. E942K mutation enhanced ERK1/2 phosphorylation without affecting ß-catenin phosphorylation. The results suggest that E942K mutation in CaSR may ultimately promote the proliferation of gastric cancer cells by enhancing intracellular calcium signaling and ERK1/2 phosphorylation. These results have potential clinical implications for the diagnosis and targeted therapy of gastric cancer.

RNA-sequencing analysis of fungi-induced transcripts from the bamboo wireworm Melanotus cribricollis (Coleoptera: Elateridae) larvae.

Larvae of Melanotus cribricollis, feed on bamboo shoots and roots, causing serious damage to bamboo in Southern China. However, there is currently no effective control measure to limit the population of this underground pest. Previously, a new entomopathogenic fungal strain isolated from M. cribricollis larvae cadavers named Metarhizium pingshaense WP08 showed high pathogenic efficacy indoors, indicated that the fungus could be used as a bio-control measure. So far, the genetic backgrounds of both M. cribricollis and M. pingshaense WP08 were blank. Here, we analyzed the whole transcriptome of M. cribricollis larvae, infected with M. pingshaense WP08 or not, using high-throughput next generation sequencing technology. In addition, the transcriptome sequencing of M. pingshaense WP08 was also performed for data separation of those two non-model species. The reliability of the RNA-Seq data was also validated through qRT-PCR experiment. The de novo assembly, functional annotation, sequence comparison of four insect species, and analysis of DEGs, enriched pathways, GO terms and immune related candidate genes were operated. The results indicated that, multiple defense mechanisms of M. cribricollis larvae are initiated to protect against the more serious negative effects caused by fungal infection. To our knowledge, this was the first report of transcriptome analysis of Melanotus spp. infected with a fungus, and it could provide insights to further explore insect-fungi interaction mechanisms.

Local injection of substance P increases bony formation during mandibular distraction osteogenesis in rats.

Substance P is a neuropeptide that is distributed in those sensory nerve fibres that innervate the medullary tissues of bone. It is a potent accelerator of proliferation and differentiation of osteoblasts in vitro. However, its capacity for promoting repair of mandibular defects is not known. We have investigated the osteogenic effects of local injections of substance P during mandibular distraction osteogenesis in rats. Twenty Sprague-Dawley rats were randomly assigned to 2 groups (n = 10 in each): substance P 10(-7) mmol/l in normal saline 0.2ml was injected into the experimental group, and saline alone into the controls. The mandibular distraction rate was 0.2mm every 12hours for 10 days. Daily injections of substance P or saline were given during the distraction period. Regeneration of bone was assessed quantitatively on days 15 and 29 using microcomputed tomography (microCT), and histological analysis. The rate of bony union in the group treated with substance P was significantly higher than that in the saline alone group on day 29 (p=0.001) The microCT images and quantitation showed more callus and more mature cortical bone when substance P was given than with control. Histological examination showed that cartilaginous tissues had formed in the middle of the distraction gaps in both groups. Bony bridges were seen only in the substance P group at the final time point (day 29). Injection of substance P into the gap of a rat mandible during mandibular distraction improved formation of good-quality bone and accelerated bony union.

[Electrophysiological and behavioral responses of male Apamea apameoides (Draudt) (Lepidoptera: Noctuidae) to sex pheromone components].

The sex pheromone gland extracts collected from calling females of Apamea apameoides (Lepidoptera: Noctuidae) were analyzed with GC-MS, the electrophysiological and behavioral responses of the male adults to serial dilutions of sex pheromone components and their synthetic blends were investigated with Y-tube olfactometer in laboratory and in bamboo forest field. The results indicated that (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol were the functional components in the sex pheromone gland extracts. Electroantennogram (EAG) recordings showed that sex pheromone gland extracts, (Z)-11-hexadecenyl acetate, (Z)-11-hexadecen-1-ol and the mixture of (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol all could elicit strong EAG responses, and the average EAG values increased with the increasing concentration of the sex pheromone. The blends of (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol at the ratio of 57:43 elicited a higher EAG value than each singular component did. The results of behavioral assay by Y-tube olfactometer accorded with those of EAG responses on the whole, and the mixture of (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol at the ratio of 57:43 was more attractive than each component alone. In field tests with silicone rubber as pheromone dispensers (concentration = 10(4) ng · uL(-1)), the average number of male adults captured per trap by the mixture was (48.5 ± 6.7).

Recruitment of exogenous mesenchymal stem cells in mandibular distraction osteogenesis by the stromal cell-derived factor-1/chemokine receptor-4 pathway in rats.

Distraction osteogenesis is widely used in orthopaedic and craniofacial surgery. However, its exact mechanism is still poorly understood. The purpose of this study was to find out whether there is systemic recruitment of mesenchymal stem cells (MSC) to the neocallus in the distraction gap by the stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis during osteogenesis. We examined the migration of MSC towards a gradient of SDF-1 in vitro. We also transplanted MSC labelled with green fluorescent protein (GFP) intravenously, with or without treatment with CXCR4-blocking antibody, into rats that had had unilateral mandibular distraction osteogenesis, and investigated the distribution of cells labelled with GFP in the soft callus after 24 h. We found that SDF-1 facilitated the migration potency of MSC both in vitro and in vivo, and this migration could be inhibited by AMD3100, an antagonist of CXCR4, and promoted by local infusion of exogenous SDF-1 into the distraction gap. This study provides a new insight into the molecular basis of how new bone is regenerated during distraction osteogenesis.

Proposal of Ensifer psoraleae sp. nov., Ensifer sesbaniae sp. nov., Ensifer morelense comb. nov. and Ensifer americanum comb. nov.

In a survey of rhizobia associated with the native legumes in Yunnan Province, China, seven and nine strains isolated from the root nodules of Psoralea corylifolia, Sesbania cannabina and Medicago lupulina were respectively classified into the novel genomic species groups I and II in the genus Ensifer (former Sinorhizobium) based on the sequence analyses of the 16S rRNA gene. Analyses of concatenated housekeeping genes (atpD, recA and glnII) further revealed that they were distinct lineages in the genus, and group I was most similar to Ensifer terangae and Ensifer garamanticus (both with 94.2% similarity), while group II was most similar to Ensifer adhaerens (94.0%). These groups could be distinguished from closely related species by DNA-DNA relatedness, MALID-TOF MS, cellular fatty acid profiles and a series of phenotypic characters. Therefore, two novel species were proposed: Ensifer psoraleae sp. nov. (seven strains, type strain CCBAU 65732(T)=LMG 26835(T)=HAMBI 3286(T)) and Ensifer sesbaniae sp. nov. (nine strains, type strain CCBAU 65729(T)=LMG 26833(T)=HAMBI 3287(T)). They had a DNA G+C mol% (Tm) of 58.9 and 60.4, respectively. Both of the type strains formed effective nodules on common bean (Phaseolus vulgaris) and their hosts of origin. In addition, the previously described species Sinorhizobium morelense and Sinorhizobium americanum were renamed as Ensifer morelense comb. nov. and Ensifer americanum comb. nov. according to the accumulated data from different studies.

A comparison of stromal cell-derived factor-1 expression during distraction osteogenesis and bone fracture in the mandible.

Distraction osteogenesis (DO) has been a widely applied technique in orthopedics and craniofacial surgery. However, the exact molecular mechanism by which the mechanical stimulus is translated into biological signals is still poorly understood. In this study, we examined and compared the expression of stromal cell-derived factor-1 (SDF-1) during mandibular distraction osteogenesis and fracture in rats, respectively. Forty-eight male Sprague-Dawley rats were divided into 2 groups and received unilateral distraction osteogenesis and rigid internal fixation, respectively, after the osteotomy on the right mandible. The harvested mandibles were examined radiographically, histologically, and immunohistochemically. We found that the expression of SDF-1 was mainly detected in the osteoblasts and blood vessels, and there were more intensive expression of SDF-1 in DO zones than in bone fracture zones. The quantitative analysis by enzyme-linked immunosorbent assay showed that SDF-1 reached a greater peak and maintained a longer period of up-regulation in DO than in fracture healing (P < 0.05). These results suggest that the distraction procedure markedly promotes the high expression of SDF-1 which facilitates the induction of bone formation during DO.

WITHDRAWN: Ensifer psoraleae sp. nov. and Ensifer sesbaniae sp. nov., isolated from the root nodules of Psoralea corylifolia, Sesbania cannabina and other legumes.

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Effects of sympathetic innervation loss on mandibular distraction osteogenesis.

Sympathetic nerve system has been proved to have important regulative effects to bone mass. However, the role of sympathetic nerve system in distraction osteogenesis (DO) is unclear. Here we show that the sympathetic nerve system plays an important role in mandibular DO. Thirty male Sprague-Dawley rats were divided into 2 groups at random. Right-side mandibular DO was performed on the 15 rats in control group (group A). Bilateral transection of cervical sympathetic trunk and right-side mandibular DO were performed on the 15 rats in the experimental group (group B). After operation, quantitative general observations, micro-computed tomography bone morphology analysis, and hematoxylin-eosin staining osseous tissue on new osteotylus in distraction gap were performed at consolidation time of 1, 14, and 28 days. SPSS 12.0 software package was used for statistical analysis. At 1 and 14 days of consolidation time, there was more continuous bone formation in the experimental group than that of the control group as determined by gross observation. Bone formation parameters including bone mineral density, bone volume-total volume ratio, bone trabeculae number as determined by micro-CT, and histological study of the test group were significantly higher than those of the control group (P < 0.05). No significant difference was noted between the 2 groups on consolidation time of 28 days. Our study suggested that the sympathetic innervation loss could improve mandibular DO and new bone formation, and the sympathetic nerve system might negatively regulate the process of DO.

[Calling and mating behaviors of bamboo shoot borer Kumasia kumaso].

The calling and mating behaviors of Kumasia kumaso were studied in laboratory at (25 1) degrees C and (60 +/- 10) % RH under a cycle 14 L:10 D photo period, and the EAG response of male K. kumaso to female K. kumaso sex gland extracts was also tested. The calling and mating behaviors of K. kumaso could only be observed in scotophase. The females began calling at the first scotophase following emergence, and the peak of calling occurred during the second scotophase and decreased thereafter. The calling of the females commenced 0-4 h after dark, reached the maximum between the 5th and the 7th hour of the scotophase, and terminated during the last 1-2.5 h of the scotophase. Moth age had significant effects on the calling patterns. With increasing moth age, the onset time of calling advanced gradually, and the mean number of calling bouts as well as the calling length per bout had a gradual increase from calling day 1 to day 4 but decreased on day 5. The peak time and terminated time of calling advanced with increasing moth age. The mating of K. kumaso adults initiated during the first scotophase and terminated till the 5th scotophase following emergence, and the peak of mating was observed during the second scotophase. The peak of mating was observed 5.5-7.0 h after dark, and advanced with increasing moth age. Moth age affected the mean onset time of mating and the copulation duration significantly. With increasing moth age, the mean onset time of mating advanced, and the copulation duration decreased. Sex ratio had significant effects on the mating behavior as well. Treatment 2 female:1 male showed a significantly higher mating percentage, but an earlier onset time of mating and shorter copulation duration, as compared with treatment 1 female:1 male. The results of EAG test indicated that the male adults showed a significant EAG response to the sex gland extracts of 2-day-old virgin females.

[Establishment of animal model of temporomandibular joint synovitis and its histological investigation].

OBJECTIVE: To establish a stable animal model of temporomandibular joint (TMJ) synovitis. METHODS: Sixteen 6-week-old male SD rats were classified into four groups, control group, occlusal dimension increase group, masseter resection group, occlusal dimension increase group and masseter resection group. The rats in the occlusal dimension increase group were adhered composite resin to their maxillary molars in order to increase the occlusal vertical dimension when they were 9-week-old. The rats in the masseter resection group were cut off their bilateral masseter muscles when they were 6-week-old. In the occlusal dimension increase group and masseter resection group, rats' bilateral masseter muscles were resected and occlusal vertical dimension was increased. All rats were sacrificed at their 10 weeks old. TMJ samples were prepared for histology to evaluate the animal model. RESULTS: The control group showed non-inflammatory changes. The occlusal dimension increase group and the masseter resection group showed vascular dilation and synovial lining proliferation, but there were no statistically significant differences between the two groups (P > 0.05). Compared to the two disposed groups, the occlusal dimension increase group and masseter resection group showed significant inflammatory changes (P < 0.05), including synovial lining proliferation, vascular dilation and fibrin deposit. CONCLUSION: The animal model of TMJ synovitis created in the present investigation could simulate the real pathological features of synovitis in vivo, and this animal model showed the obvious merits of high stability and reproduction.