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Heterostyly is a floral polymorphism characterized by reciprocal herkogamy maintained through high levels of mating between morphs, serviced by appropriate pollinators. We studied how differential efficiency and abundance of distinct pollinators affect plant female reproduction in self- and intra-morph incompatible distylous Primula secundiflora. Bumblebees and syrphid flies were found to be the most abundant floral visitors. Bumblebees frequently exhibited nectar-robbing behavior. Because the robbing holes were always situated between the high- and low-level organs on both morphs, nectar-robbing bumblebees only pollinated S-styled flowers. L-styled flowers set four times as many seeds as did S-styled flowers after being visited by pollen-collecting syrphid flies. The natural female fecundity and the magnitude of pollen limitation varied between the morphs within populations because of the mosaic distribution of nectar-robbing bumblebees and syrphid flies. L-styled flowers and S-styled flowers set the same number of seeds after supplemental hand pollination, indicating equivalent female reproductive potential. We suggest that bumblebee nectar robbers and syrphid flies play an important role in sustaining the floral dimorphism of heterostyly in P. secundiflora because of their complementary roles in the pollination system. [ABSTRACT FROM AUTHOR]

Techniques of reduced‐representation sequencing (RRS) have revolutionized ecological and evolutionary genomics studies. Precise establishment of orthologs is a critical challenge for RRS, especially when a reference genome is absent. The proportion of shared heterozygous sites across samples is an alternative criterion for filtering paralogs. In the prevailing pipeline for variant calling of RRS data – PYRAD/IPYRAD, maxSH is an often overlooked parameter with implications to detecting and filtering paralogs according to shared heterozygosity. Using empirical genotyping by sequencing data of two primroses (Primula alpicola Stapf and Primula florindae Ward) and their putative hybrids, and extra data sets of Californian golden cup oaks, we explore the impact of maxSH on filtering paralogs and further downstream analyses. Our study sheds light on the simultaneous validity and risk of filtering paralogs using maxSH, and its significant effects on downstream analyses of outlier detection, population assignment, and demographic modeling, emphasizing the importance of attention to detail during bioinformatic processes. The mutual confirmation between results of population assignment and demographic modeling in this study suggested maxSH = 0.10 has a potentially excessive and asymmetrical effect on the removal of truly shared heterozygous sites as paralogs. These results indicate that hybridization origin hypotheses of putative hybrids represented by results with maxSH = 0.25 and 0.50 are more credible. In conclusion, we revealed the critical hazard of paralogs filtration according to sharing heterozygosity at first, so that we propose to use specific protocols, rather than maxSH, to filter potential paralogs for closely related lineages. [ABSTRACT FROM AUTHOR]

Roscoea stenophylla Y. L. Fan, a new species of the ginger family (Zingiberaceae) from Yunnan, China, is described. It morphologically resembles R. schneideriana (Loesener) Cowley in leaf shape, individual size, inflorescence shape and floral color, but is significantly distinguished by longer and narrower leaves, longer corolla tubes, reflexed labellum, inflated and not hooked stigma, and anther bent into a semicircle (U type) without inflation at the end of the connective spurs. [ABSTRACT FROM AUTHOR]

Osteoarthritis is a degenerative joint disease that affects the aging population worldwide. It has an underlying inflammatory cause that leads to loss of chondrocytes, reducing the cartilage layer at the affected joints. Compounds with anti-inflammatory properties are potential therapeutic agents for osteoarthritis. Curcumin, derived from species of the Curcuma, is an anti-inflammatory compound. The purpose of this review is to summarize the anti-osteoarthritic effects of curcumin from clinical and preclinical studies. Many clinical trials have been conducted to determine curcumin's effectiveness in osteoarthritis patients. Available studies have shown that curcumin prevents chondrocyte apoptosis and inhibits the release of proteoglycans and metalloproteinases as well as the expression of cyclooxygenase, prostaglandin E-2, and inflammatory cytokines in chondrocytes. The mechanism of action of curcumin also involves multiple cell signaling pathways, including Nuclear factor kappa-B(NF-κB), Mitogen-activated protein kinase (MAPK), Wnt/β-catenin pathway (Wnt/β-catenin), The Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3), Nuclear factor erythroid 2-related factor 2/antioxidant response elements/heme oxygenase-1(Nrf2/ARE/HO-1), and Phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling pathways. Curcumin further reduced the release of inflammatory factors and apoptosis by inhibiting the activation of NF-κB. In addition, curcumin modulates the MAPK, Nrf2/ARE/HO-1, and PI3K/Akt/mTOR signaling pathways and affects cell proliferation and apoptosis processes, a series of effects that together promote the healthy state of chondrocytes. In conclusion, curcumin, as a natural plant compound, exhibits significant anti-inflammatory potential by modulating inflammatory factors associated with articular osteoarthritis through multiple mechanisms. Its protective effects on articular cartilage and synovium make it a promising candidate for the treatment of OA. Future studies should further explore the mechanism of action of curcumin and its optimal dosage and therapeutic regimen in clinical applications, to provide more effective therapeutic options for osteoarthritis patients. [ABSTRACT FROM AUTHOR]

The homogeneous balance of undetermined coefficients method is proposed to obtain not only exact solutions but also multi-symplectic structure of some nonlinear partial differential equations. Bilinear equation, N-soliton solutions, traveling wave solutions and multi-symplectic structure are obtained by applying the proposed method to the KdV equation. Accordingly, the definition and multi-symplectic structure of the generalized KdV-type equation are given. The proposed method is also a standard and computable method, which can be generalized to deal with some types of nonlinear partial differential equations. [ABSTRACT FROM AUTHOR]

Premise: Spatial and temporal resource allocations within inflorescences have been well‐studied in many plants based on flowering sequence or floral position. However, there had been few attempts to investigate architectural effects and resource competition in species where the blooming pattern does not follow a linear positional pattern within the inflorescence. Moreover, most flowering plants show female‐biased sex allocation in early or basal flowers, but it is unclear in species with inherent and changeless ovule production. Methods: We investigated intra‐inflorescence variation in reproductive traits of Salvia przewalskii, a perennial herb with 4‐ouvle ovary flowers and flowering sequence–floral position decoupled inflorescences. To detect the effects of resource competition and architectural effects on reproductive success, we manipulated inflorescence (removed floral buds by position and flowering sequence) and pollination (opened and supplemented pollination). Results: Pollen production and dry mass deceased from bottom to top flowers but did not significantly differ following flowering sequence, resulting in male‐biased sex allocation in basal flowers. The seed production, fruit set, and bud development exhibited significant declining trends from proximal to distal positions regardless of the thinning and pollen treatments. Meanwhile, the seed production, fruit set, and bud development success did not significant differ when thinning was conducted according to flowering sequence. Conclusions: Architectural effects plays a crucial role in resource allocation within decoupled flowering inflorescences. Moreover, our results highlighted that inherent floral traits such as changeless ovule production, may modify architectural effects on sex allocation. [ABSTRACT FROM AUTHOR]

Aim: The relationships between biome shifts and global environmental changes in temperate zone habitats have been extensively explored; yet, the historical dynamics of taxa found in the tropical rain forest (TRF) remain poorly known. This study aims to reconstruct the relationships between tropical rain forest shifts and global environmental changes through the patterns of historical biogeography of a pantropical family of monocots, the Zingiberaceae. Location: Global. Taxon: Zingiberaceae. Methods: We sampled DNA sequences (nrITS, trnK, trnL‐trnF and psbA‐trnH) from GenBank for 77% of the genera, including 30% of species, in the Zingiberaceae. Global fossil records of the Zingiberaceae were collected from literatures. Rates of speciation, extinction and diversification were estimated based on phylogenetic data and fossil records through methods implemented in BAMM. Ancestral ranges were estimated using single‐tree BioGeoBEARS and multiple‐trees BioGeoBEARS in RASP. Dispersal rate through time and dispersal rate among regions were calculated in R based on the result of ancestral estimation. Results: The common ancestor of the Zingiberaceae likely originated in northern Africa during the mid‐Cretaceous, with later dispersal to the Asian tropics. Indo‐Burma, rather than Malesia, was likely a provenance of the common ancestor of Alpinioideae–Zingiberoideae. Several abrupt shifts of evolutionary rates from the Palaeocene were synchronized with sudden global environmental changes. Main conclusions Integrating phylogenetic patterns with fossil records suggests that the Zingiberaceae dispersed to Asia through drift of the Indian Plate from Africa in the late Palaeocene. Formation of island chains, land corridors and warming temperatures facilitated the emigration of the Zingiberaceae to a broad distribution across the tropics. Moreover, dramatic fluctuations of the speciation rate of Zingiberoideae appear to have been synchronized with global climate fluctuations. In general, the evolutionary history of the Zingiberaceae broadens our understanding of the association between TRF shifts in distribution and past global environmental changes, especially the origin of TRF in Southeast Asia. [ABSTRACT FROM AUTHOR]

Background: The Notch signaling pathway plays an important role in cancer, but the mechanism by which Notch1 participates in invasion and migration of hepatocellular carcinoma (HCC) cells is unclear. Aims: Our purpose is to confirm the anti-invasion and anti-migration effects of the down-regulation of Notch1 in HCC cells. Methods: The invasion and migration capacities of HCC cells were detected with Transwell cell culture chambers. The expressions of Notch1, Notch1 intracellular domain (N1ICD), E-cadherin, Snail, and cyclooxygenase-2 (COX-2) were analyzed by RT-PCR and/or western blotting. Notch1 and Snail were down-regulated by RNA interference, and COX-2 was inhibited by NS-398. Cell apoptosis was analyzed by MTT and flow cytometry. Results: In HCC cells, Snail, Notch1, and COX-2 were up-regulated, and E-cadherin was down-regulated in mRNA and/or protein levels. The down-regulation of Snail or Notch1 or the inhibition of COX-2, respectively, can increase the mRNA and protein expressions of E-cadherin and decrease the invasion and migration capabilities of HCC cell. Down-regulated Notch1 or inhibited COX-2 can reduce the mRNA and protein expressions of Snail. The down-regulation of Notch1 can also reduce the protein expression of COX-2. However, exogenous PGE2 can reverse the role of down-regulated Notch1. The results of MTT and flow cytometry showed that down-regulated Notch1 did not affect HCC cell viability. Conclusions: Down-regulated Notch1 may be an effective approach to inactivating Snail/E-cadherin by regulating COX-2, which results in inhibiting the invasion and migration of HCC cells. The inhibitory effects of down-regulated Notch1 on cell invasion and migration were independent of apoptosis. [ABSTRACT FROM AUTHOR]

Aim: Adaptation, migration and extinction of species is closely associated with climate changes and shape the distribution of biodiversity. The adaptive responses of species in the biodiversity hotspot, the Hengduan Mountains, to climate change remain poorly understood. Location: The Hengduan Mountains, southeast of the Qinghai‐Tibet Plateau. Methods: The principles of phylogeography and landscape genomics were applied to the endemic species Roscoea humeana in the Hengduan Mountains by genotyping by sequencing data. A total of 5,902 single‐nucleotide polymorphisms were used to analyse the genetic structure/diversity and changes in effective population size over time. Species distribution modelling, principal component analysis and gradient forest analysis were used to explore associations between climate change and genetic variation. Results: The population size of R. humeana rapidly increased after the Last Glacial Maximum. The contribution of climate to genetic variation was greater than that of geography. Precipitation during the warmest season played a pivotal role in the adaptation to climate changes. Loci associated with drought tolerance and anti‐ultraviolet radiation were identified, suggesting local adaptation of R. humeana to alpine environments. Main conclusions: These results suggest that the current genetic structure and diversity of R. humeana were shaped by Quaternary climate fluctuations and persistence of microrefugia in the Hengduan Mountains. The results further suggest that R. humeana can survive in future refugia within the area where the warmest quarter precipitation is higher than 560 mm, and local adaptation to drought tolerance may be beneficial for its acclimation to future climate changes. [ABSTRACT FROM AUTHOR]

Keywords: alpine ginger; autonomous selfing; pollination; pollinator failure; reproductive assurance; Roscoea nepalensis; stigmatic droplet EN alpine ginger autonomous selfing pollination pollinator failure reproductive assurance Roscoea nepalensis stigmatic droplet 1 5 5 07/05/21 20210701 NES 210701 The genus I Roscoea i , a Himalayan endemic and only alpine member of the predominately tropical family Zingiberaceae, is well-known for possessing elaborate floral traits commonly associated with biotic pollination, and particularly with long-tongued fly pollination. Our current findings on the pollination mechanism of I R. nepalensis i , i.e., the occurrence of dual autonomous pollination mechanisms, with the unusual use of two mechanisms distributed among different flowers, during the potentially competing stage is new to the broad range of pollination mechanisms evolved in angiosperms. Consistent with these floral traits, most I Roscoea i spp. exhibit either obligate or partial biotic pollination (Zhang et al. 2010, Fan and Li 2012, Paudel et al. 2015, 2017, 2019), while entirely autonomous self-pollination is rare (Zhang and Li 2008). [Extracted from the article]

Sexual interference between male and female function in hermaphrodite plants is reduced by protandry. In environments with insufficient pollinator service, prolongation of male function owing to limited pollen removal could restrict the duration of female function and lower seed production. We provide evidence that this form of sexual conflict has played a role in the spread of females in gynodioecious populations of Cyananthus delavayi in the pollen‐limited environments in which this subalpine species occurs. Using field experiments involving artificial pollen removal from the strongly protandrous flowers of hermaphrodites, we demonstrated a trade‐off between male‐ and female‐phase duration with no influence on overall floral longevity. Pollen removal at the beginning of anthesis resulted in hermaphrodite seed production matching that of females. In contrast, restricted pollen removal increased the duration of male function at the expense of female function lowering maternal fertility compared to females. This pattern was evident in five populations with females experiencing a twofold average seed fertility advantage compared to hermaphrodites. Gynodioecy often appears to evolve from protandrous ancestors and pollen limitation is widespread in flowering plants suggesting that sexual conflict may play an unappreciated role in the evolution of this form of sexual dimorphism. [ABSTRACT FROM AUTHOR]

Summary: Water‐mediated fertilization is ubiquitous in early land plants. This ancestral mode of fertilization has, however, generally been considered to have been lost during the evolutionary history of terrestrial flowering plants.We investigated reproductive mechanisms in the subtropical ginger Cautleya gracilis (Zingiberaceae), which has two pollen conditions – granular and filiform masses – depending on external conditions. We tested whether rain transformed granular pollen into filiform masses and whether this then promoted pollen‐tube growth and fertilization of ovules. Using experimental manipulations in the field we investigated the contribution of water‐mediated fertilization to seed production.Rain caused granular pollen to form filiform masses of germinating pollen tubes, which transported sperm to ovules, resulting in fertilization and seed set. Flowers exposed to rain produced significantly more seeds than those protected from the rain, which retained granular pollen. Insect pollination made only a limited contribution to seed set because rainy conditions limited pollinator service.Our results reveal a previously undescribed fertilization mechanism in flowering plants involving water‐mediated fertilization stimulated by rain. Water‐mediated fertilization is likely to be adaptive in the subtropical monsoon environments in which C. gracilis occurs by ensuring reproductive assurance when persistent rain prevents insect‐mediated pollination. [ABSTRACT FROM AUTHOR]

Aim: Ecological differentiation of closely related species has attracted wide attention to explore its evolutionary significance in speciation. In particular, the prevalence of ecological speciation of sister species driven by Quaternary climate changes is debated. Here, we used two parapatric sister species, Roscoea humeana and R. cautleoides, to test whether ecological speciation was associated with Quaternary climate changes in these taxa. Location: Hengduan-Mountain Region (HMR), south-west of China. Methods: Nuclear ribosomal (ITS) and chloroplast DNA (cpDNA: psbA-trnH and trnL-trnF) were sequenced from 200 individuals. Gene genealogies were reconstructed using network, Bayesian inference and maximum likelihood methods. Interspecific divergence times were estimated with an isolation with migration model. Suitable habitats for each species were estimated with ecological niche modelling using Maxent and BIOCLIM. Niche overlap and niche divergence were tested using Schoener's D and Warren's I. Principal components analysis (PCA), multivariate analysis of variance (MANOVA) and discriminant function analysis (DFA) were also used to test ecological differentiation. Results: Gene genealogies indicated that the low-elevational species R. cautleoides was derived from the high-elevational species R. humeana. Divergence among species occurred during the middle to late Quaternary (0.01-1.48 Ma). The species distribution and phylogeographical pattern were inconsistent with a geographical barrier. Niche models and statistical analyses showed significant ecological differentiation. Main conclusions: Divergence between R. humeana and R. cautleoides was recent and coincident with Quaternary climate cycles. Ecological niche differentiation within Quaternary refugia may have promoted speciation. [ABSTRACT FROM AUTHOR]

Background and Aims Flower morphology and inflorescence architecture affect pollinator foraging behaviour and thereby influence the process of pollination and the reproductive success of plants. This study explored possible ecological functions of the lever-like stamens and the floral design in Salvia cyclostegia. Methods Flower construction was experimentally manipulated by removing either the lower lever arms or the upper fertile thecae of the two stamens from a flower. The two types of manipulated individuals were intermixed with the control ones and randomly distributed in the population. Key Results Removing the sterile lower lever arms significantly reduced handling time per flower of the main pollinator, Bombus personatus. Interestingly, this manipulation did not increase the number of flowers probed per plant visit, but instead reduced it, i.e. shortened the visit sequence of the bumble-bees. Both loss of staminal lever function by removing lower lever arms and exclusion of self pollen by removing upper fertile thecae significantly reduced seed set per flower and seed set per plant. Both the manipulations interacted significantly with inflorescence size for the effect on female reproductive output. Conclusions Though the intact flowers demand a long handling time for pollinators, the reversible staminal lever is of advantage by promoting dispersal of pollen and thus the male function. The particular floral design in S. cyclostegia contributes to the floral constancy of B. personatus bumble-bees, with the lower lever arms acting as an optical cue for foraging cognition. [ABSTRACT FROM PUBLISHER]

Abstract: This study investigates whether activation of p38 MAPK by the up-regulation of HSP 70 participates in the induction of brain ischemic tolerance by limb ischemic preconditioning (LIP). Western blot and immunohistochemical assays indicated that p38 MAPK activation occurred earlier than HSP 70 induction in the CA1 region of the hippocampus after LIP. P-p38 MAPK expression was up-regulated at 6h and reached its peak 12h after LIP, while HSP 70 expression was not significantly increased until 1day and peaked 2days after LIP. Neuropathological evaluation by thionin staining showed that quercetin (4ml/kg, 50mg/kg, intraperitoneal injection), an inhibitor of HSP 70, blocked the protective effect of LIP against delayed neuronal death that is normally induced by lethal brain ischemic insult, indicating that HSP 70 participates in the induction of brain ischemic tolerance by LIP. Furthermore, SB 203580, an inhibitor of HSP 70, inhibited HSP 70 activation in the CA1 region of the hippocampus induced by LIP either with or without the presence of subsequent brain ischemic insult. Based on the above results, it can be concluded that activation of p38 MAPK participates in the brain ischemic tolerance induced by LIP at least partly by the up-regulation of HSP 70 expression. [Copyright &y& Elsevier]

Abstract: Little information is currently available regarding emissions of biogenic volatile organic compounds (BVOCs) in southern Asia. To address the need for BVOC emission estimates in regional atmospheric chemistry simulations, 95 common plant species were screened for emissions of BVOC in and near the Xishuangbanna Tropical Biological Gardens in southern Yunnan Province, Peoples’ Republic of China in February 2003. In situ measurements with leaf cuvettes and branch bag enclosures were used in combination with portable gas chromatography, flame ionization, photoionization, and mass spectral detection to identify and quantify BVOC emissions. Forty-four of the species examined emitted isoprene at rates exceeding 20μg Cg−1 (leaf dry weight) h−1. An emphasis was placed on the genus Ficus, which is important in the region and occupies a wide range of ecological niches. Several species in the footprint of a nearby flux tower were also examined. Several palm species and an abundant fern (Cyclosorus parasiticus) emitted substantial amounts of isoprene, and probably accounted for observed daytime mean isoprene fluxes from the understory of a Hevea brasiliensis plantation of 1.0 and 0.15mgCm−2 h−1 during the wet and dry seasons, respectively. These measurements verify that both the forest floor and canopy in this region can be sources of isoprene. Monoterpene emissions exceeded 1.0μg-Cg−1 (leaf dry weight) h−1 from only 4 of 38 species surveyed, including some Ficus species and H. brasiliensis. However most of the trees of the latter species were sparsely foliated due to dry season senescence, and emission factors are approximately an order of magnitude lower than those reported during the wet season. BVOC emission rates and physiology of many species are impacted by reduced moisture availability, especially Mangifera indica. South Asia is a region undergoing rapid landuse change and forest plantation establishment, with large increases in area of high BVOC-emitting species in the genera Bambusa, Elaeis, Eucalyptus, Hevea, Pinus, and Populus (among others). This could result in profound changes in atmospheric chemistry in these regions, for instance, terpene emissions from H. brasiliensis could increase wet season biogenic organic aerosol burdens by approximately a factor of 2 in the Xishuangbanna region. Increases in plantation area established with high isoprene emitting species, (e.g. Bambusa spp. and Eucalyptus spp.) are also projected for China and other parts of Southeast Asia in the near future. Thus, landcover change in South Asian landscapes is usually associated with large increases in BVOC flux with the potential to alter the atmospheric chemical composition and air quality over this rapidly developing region. [Copyright &y& Elsevier]

Abstract: Canopy scale fluxes of isoprene and monoterpenes were investigated in both wet and dry seasons above a rubber tree (Hevea brasiliensis)/secondary tropical forest in the Yunnan province of southwestern China. Drought conditions were unusually high during the dry season experiment. The eddy covariance measurement technique was used to measure isoprene fluxes, while monoterpene fluxes were modeled based on leaf level emission measurements. Maximum observed isoprene fluxes occurred during the wet season and daytime average fluxes were about 1mg Cm-2h-1. Dry season fluxes were much lower with a daytime average of 0.15mg Cm-2h-1. Wet season isoprene fluxes compare quite well with isoprene fluxes observed from other tropical forests. Monoterpene fluxes came, almost entirely, from Hevea brasiliensis, which is a light-dependent monoterpene emitter. Modeled wet season total monoterpene fluxes were about 2mg Cm-2h-1 (average for the daytime), and in the dry season were undetectable. Extreme drought conditions, and the drought deciduous nature of Hevea brasiliensis may be the cause of the low dry season fluxes. [Copyright &y& Elsevier]

[Display omitted] • Clear phylogenetic relationships were reconstructed by 32,375 unlinked SNPs from restriction site-associated DNA sequencing. • The first divergence of Roscoea was the result of vicariance driven by the rapid uplift of the Himalayas. • Speciation occurred from highland toward lowland driven by climate cooling after the first divergence. • Interspecific introgressions are ancient and no introgression among extant species. • This study has significant contributions to understand the causes of extraordinary high mountain biodiversity. The Himalayas with dramatic elevation gradient is one of the global biodiversity hotspots. Although origin of biodiversity of the Himalayas is of great concern, the speciation process within the Himalayas is poorly known. Roscoea within the Himalayas serve as a good model system to test the speciation process along an elevation gradient. 32,375 unlinked SNPs were used to reconstruct phylogenetic relationships and introgression analyses in D -statistics and Fastsimicoal 2. Species distribution modeling (SDM) was used to simulate habitat shift of Roscoea species during climate changes. Phylogeny suggested that the speciation order, except R. capitata , was from highland to lowland. D -statistics analyses suggested significant bidirectional ancient introgression between elevation-neighboring clades but no introgression between R. capitata and othern clades and no introgression among extant species. Fastsimicoal 2 suggested interspecific introgressions were asymmetric. SDM predicted that habitats of Roscoea shifted to low elevation during cooling age. These results suggested that the sudden uplift of the Himalayas likely promoted speciation by vicariance, and climate cooling drove species divergence towards lower elevation. This study provides explanations for the origin of biodiversity within the Himalayas, and an insight to understand speciation along elevation in the mountainous regions. [ABSTRACT FROM AUTHOR]

Objective: To explore the potential mechanism of lysionotin in treating glioma., Methods: First, target prediction based on Bernoulli Naïve Bayes profiling and pathway enrichment was used to predict the biological activity of lysionotin. The binding between 5-lipoxygenase (5-LO) and lysionotin was detected by surface plasmon resonance (SPR) and molecular docking, and the inhibitory effects of lysionotin on 5-LO and proliferation of glioma were determined using enzyme inhibition assay in vitro and cell viability analysis, respectively. Furthermore, the pharmaceutical effect of lysionotin was explored by cell survival rate analysis and liquid chromatography with tandem mass spectrometry (LC-MS/MS). The protein expression, intracellular calcium ion concentration and cytoskeleton detection were revealed by Western blot, flow cytometry and fluorescence labeling, respectively., Results: Target prediction and pathway enrichment revealed that lysionotin inhibited 5-LO, a key enzyme involved in the arachidonic acid metabolism pathway, to inhibit the proliferation of glioma. Molecular docking results demonstrated that 5-LO can be binding to lysionotin through hydrogen bonds, forming bonds with His600, Gln557, Asn554, and His372. SPR analysis further confirmed the interaction between 5-LO and lysionotin. Furthermore, enzyme inhibition assay in vitro and cell survival rate analysis revealed that 50% inhibition concentration of lysionotin and the median effective concentration of lysionotin were 90 and 16.58 µmol/L, respectively, and the results of LC-MS/MS showed that lysionotin inhibited the production of 5S-hydroperoxy-eicosatetraenoic acid (P<0.05), and moreover, the LC-MS/MS results indicated that lysionotin can enter glioma cells well (P<0.01) and inhibit their proliferation. Western blot analysis demonstrated that lysionotin can inhibit the expression of 5-LO (P<0.05) and downstream leukotriene B4 receptor (P<0.01). In addition, the results showed that lysionotin affected intracellular calcium ion concentration by inhibiting 5-LO to affect the cytoskeleton, as determined by flow cytometry and fluorescence labeling., Conclusion: Lysionotin binds to 5-LO could suppress glioma by inhibiting arachiodonic acid metabolism pathway., (© 2024. The Chinese Journal of Integrated Traditional and Western Medicine Press and Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Pancreatic cancer (PC) is the most lethal malignant tumor, with average survival period of about 10 months. C-X-C ligand 5 (CXCL5), an important chemokine for immune cell accumulation in tumor tissues, has been reported to be involved in a variety of human cancers. However, the exact role of CXCL5 in PC progression has not been well defined., Methods: The expression of CXCL5 in PC was analyzed based on online databases and clinical specimens immunohistochemical staining, and Western blotting of CXCL5 in PC cell lines and patient samples. The correlation between CXCL5 expression and prognosis in PC was explored. The role of CXCL5 in PC was investigated through in vitro and in vivo experiments., Results: The expression of CXCL5 was significantly increased in PC tissues compared with that in pancreas tissues, and CXCL5 high expression predicts poor prognosis in PC patients. Further analyses demonstrated that overexpression of CXCL5 in PC cells was positively related to higher proliferation rate, higher migration ability, and higher EMT markers including SNAI2 and TWIST1 of tumor cells in vitro. Consistently, the knockdown of CXCL5 in PC cells harmed the proliferation rate, migration ability, and expression of EMT indexes of tumor cells in vitro. Importantly, knockdown of CXCL5 inhibited the growth of xenograft tumors in vivo., Conclusion: CXCL5 high expression predicts poor prognosis in PC patients. CXCL5 promotes PC cell growth and EMT process. Inhibition of CXCL5 may be a potential therapeutic approach for PC., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Paraneoplastic neurological syndrome (PNS) is an unusual event. PNS caused by cystitis glandularis (CG) or a bladder tumor is extremely rare; hence, missed diagnosis or misdiagnosis can easily occur. To date, approximately 21 cases have been reported in PubMed., Case Summary: We report a case of PNS caused by CG and describe the clinical and imaging features. The main clinical feature was advanced cognitive impairment, and early clinical features were memory impairment, decreased computational ability, and abnormal behavior. Later clinical features were dementia, vomiting, inability to eat and walk, urinary incontinence, and hematuria. Imaging features on cranial magnetic resonance imaging were diffuse white matter lesions. Paraneoplastic tumor markers were normal. A total abdominal computed tomography scan showed multiple thickened areas on the bladder wall with local prominence. Cystoscopy revealed a volcanic protuberance on the posterior wall of the bladder with a diameter of 6 cm and no pedicle. The postoperative pathological diagnosis was CG. The patient recovered well following resection of CG. PNS cases caused by previous bladder tumors can be retrieved from PubMed to describe the clinical signs and prognosis of PNS., Conclusion: The main clinical feature of PNS caused by CG was dementia, and the imaging features were diffuse cerebral white matter lesions. Resection of CG lesions is the fundamental treatment for PNS induced by CG. This case highlights the importance of etiological treatment., Competing Interests: Conflict-of-interest statement: All authors report no relevant conflict of interest for this article., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Chloroplasts are critical to plant survival and adaptive evolution. The comparison of chloroplast genomes could provide insight into the adaptive evolution of closely related species. To identify potential adaptive evolution in the chloroplast genomes of four montane Zingiberaceae taxa ( Cautleya , Roscoea , Rhynchanthus , and Pommereschea ) that inhabit distinct habitats in the mountains of Yunnan, China, the nucleotide sequences of 13 complete chloroplast genomes, including five newly sequenced species, were characterized and compared. The five newly sequenced chloroplast genomes (162,878-163,831 bp) possessed typical quadripartite structures, which included a large single copy (LSC) region, a small single copy (SSC) region, and a pair of inverted repeat regions (IRa and IRb), and even though the structure was highly conserved among the 13 taxa, one of the rps19 genes was absent in Cautleya , possibly due to expansion of the LSC region. Positive selection of rpoA and ycf2 suggests that these montane species have experienced adaptive evolution to habitats with different sunlight intensities and that adaptation related to the chloroplast genome has played an important role in the evolution of Zingiberaceae taxa., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Yang, Fu, Wu, Li, Zhao and Li.)

The population characteristics of distylous species are highly sensitive to stochastic natural selection pressure. Therefore, populations growing under different environmental conditions may vary in floral morph ratios, potentially affecting female fitness and leading to inbreeding depression. However, the variation in offspring quality among populations as a result of inbreeding depression is poorly understood in distylous species. This study investigates variations in plant density, seed mass, seed viability, female fitness, and post-dispersal inbreeding depression in both sexual morphs (long-styled and short-styled plants) of the distylous Primula nivalis that were subjected to different pollination treatments along an elevational gradient from 1657 to 2704 m a.s.l. Population characteristics (morph plant density and ratio) and fruit set were significantly affected by sexual morph and elevation. Plant density and fruit set frequencies were lower for short-styled than for long-styled plants at 2704 m a.s.l. The seeds from the cross-pollinated flowers of both morphs were higher in quality than those of self-pollinated flowers. The female fitness of seeds from cross-pollinated flowers of both morphs was higher than that of seeds from open-pollinated and self-pollinated flowers. The female fitness of seeds from long-styled flowers was higher than that of seeds from short-styled flowers at all elevations. Inbreeding depression increased with elevation among plants with short-styled flowers but not among those with long-styled flowers. Variation in the elevation-dependent mating system might influence female fitness and affect inbreeding depression in both floral morphs. In conclusion, the low quality of seeds from short-styled flowers at high elevations might decrease short-styled flower frequency, affecting population characteristics.

Influenza A virus infection represents a great threat to public health. However, owing to side effects and the emergence of resistant virus strains, the use of currently available anti-influenza drugs may be limited. In order to identify novel anti-influenza drugs, we investigated the antiviral effects of phillyrin against influenza A virus infection in vivo. The mean survival time, lung index, viral titers, influenza hemagglutinin (HA) protein and serum cytokines levels, and histopathological changes in lung tissue were examined. Administration of phillyrin at a dose of 20 mg/kg/day for 3 days significantly prolonged the mean survival time, reduced the lung index, decreased the virus titers and interleukin-6 levels, reduced the expression of HA, and attenuated lung tissue damage in mice infected with influenza A virus. Taken together, these data showed that phillyrin had potential protective effects against infection caused by influenza A virus.

Due to invasion and intrahepatic metastasis, the prognosis for patients with hepatocellular carcinoma (HCC) is poor. However, the mechanisms underlying these processes of HCC remain unclear. Cancer stem cells may be involved in early systemic dissemination and metastasis formation and side population (SP) cells isolated from diverse cancer cells possess stem cell-like properties. However, the mechanisms involved in migration and invasion of cancer stem cells are not well understood. In this study, we identified and isolated populations of SP cells from HCC cell lines using flow cyto-metry. SP cells showed higher levels of migration and invasion capability. Higher expression of miR-21 was observed in SP cells. Silencing of miR-21 led to a reduction in the migration and invasion of these cells and overexpression of miR-21 can increase in cell migration and invasion. Overexpression of miR-21 did not cause degradation of PTEN or RECK or PDCD4 mRNA but drastically inhibited its protein expression. Consistent with these results, silencing miR-21 increased the levels of PTEN, RECK and PDCD4 protein, respectively. The role of silencing miR-21 was partially attenuated by silencing of PTEN or RECK or PDCD4 mRNA. The results of this study revealed the aberrant expression of miR-21 in SP cells and showed that miR-21 regulates the expression of multiple target proteins that are associated with tumor dissemination. MiR-21 is a pro-metastatic miRNA in SP cells and raises the possibility that therapy of HCC may be improved by pharmaceutical strategies directed towards miR-21.

Although Notch1 expression has been associated with progression or prognosis in various tumors, the role of Notch1 in hepatocellular carcinoma (HCC) remains unknown. This study sought to investigate the clinicopathological and prognostic relevance of Notch1 expression in HCC as well as the underlying mechanisms responsible. HCC tissues were stained with an anti-Notch1 antibody. The invasion capacities of cells were measured using Transwell cell culture chambers. Reverse transcription PCR and/or western blot were used to evaluate the expression levels of Notch1, matrix metalloproteinase (MMP)-2, and MMP-9. Notch1 expression was downregulated by RNA interference. The activity of MMP-2/MMP-9 was quantified by enzyme-linked immunosorbent assay, and cellular apoptosis was analyzed using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Notch1 expression was mainly localized within the cytoplasm and at the cell membrane. High Notch1 expression correlated with tumor size, tumor grade, metastasis, venous invasion, and American Joint Committee on Cancer TNM stage (P < 0.05), and patients with high levels of Notch1 expression were at a significantly increased risk for shortened survival time (P < 0.05). In vitro, the downregulation of Notch1 expression decreased the invasion capacity of HCC cells via the regulation of MMP-2 and MMP-9. The results of the MTT assay showed that downregulation of Notch1 did not affect HCC cell viability. Notch1 may represent a novel candidate marker for patient prognosis as well a molecular target for HCC therapy.

Objective: To investigate the role of heme oxygenase and carbon monoxide (HO/CO) in the development of spontaneous pain and hyperalgesia of rats induced by formalin injection., Methods: Zinc protoporphyrin Znpp (the inhibitor of HO) was intrathecally injected to the rats with formalin inflammatory pain. Hemin (the agonist of HO) was intrathecally injected to the normal rats. The weighted pain scores were used to evaluate the degree of pain response. Thermal withdrawal latency and mechanical withdrawal threshold were observed to assess the degree of thermal hyperalgesia and mechanical allodynia., Results: After the intrathecal injection of Znpp, the weighted pain score obviously reduced in a dose-dependent manner compared with the rats with formalin inflammatory pain. Intrathecal injection of Znpp had no obvious effect on thermal withdrawal latency and mechanical withdrawal threshold in injected feet compared with formalin group. But there was a prolongation in a dose-dependent manner in non injected feet. Intrathecal injection of Hemin to normal rats could shorten the thermal withdrawal latency and reduce the mechanical withdrawal threshold on both sides of hindpaws., Conclusion: Intrathecal injection of the HO inhibitor produced prominent inhibition to pain related behavior and thermal and mechanical hyperalgesia induced by formalin injection. Intrathecal injection of HO inductor could induce thermal and mechanical hyperalgesia in normal rats. The results indicated that HO/CO took part in the processes of spinal cord nociceptive information transmission and the development of thermal and mechanical hyperalgesia.

MicroRNA-10b (miR-10b) was recently reported to be dysregulated in some types of cancer and to play a role in invasion and metastasis. However, effects and potential mechanisms of action of miR-10b in the metastasis of hepatocellular carcinoma (HCC) have not been explored. In this study, we confirmed that miR-10b is highly expressed in metastatic HCC tissues and in metastatic HCC cell lines by qRT-PCR. Moreover, patients with higher miR-10b expression had significantly poorer overall survival, and high miR-10b expression was an independent predictor of poor prognosis. Inhibition of miR-10b reduced cell migration and invasion in MHCC97H cells, whereas over-expression of miR-10b in HepG2 cells increased cell migration and invasion. Bioinformatics and luciferase reporter assays revealed that miR-10b binds the 3'-UTR of CADM1 mRNA and represses its translation. Western blot and qRT-PCR showed that CADM1 is inhibited by miR-10b over-expression. Silencing of CADM1 resulted in substantially increased cell motility and invasion similar to that observed with over-expression of miR-10b in HepG2 cells. These results suggest that miR-10b may positively regulate the invasion and metastasis of HCC through targeting CADM1.

Background and Aims: Delayed selfing is the predominant mode of autonomous self-pollination in flowering plants. However, few delayed selfing mechanisms have been documented. This research aims to explore a new delayed selfing mechanism induced by stigmatic fluid in Roscoea debilis, a small perennial ginger., Methods: Floral biology and flower visitors were surveyed. The capacity of autonomous selfing was evaluated by pollinator exclusion. The timing of autonomous selfing was estimated by emasculation at different flowering stages. The number of seeds produced from insect-pollination was assessed by emasculation and exposure to pollinators in the natural population. The breeding system was also tested by pollination manipulations., Key Results: Autonomous self-pollination occurred after flowers wilted. The stigmatic fluid formed a globule on the stigma on the third day of flowering. The enlarged globule seeped into the nearby pollen grains on the fourth flowering day, thus inducing pollen germination. Pollen tubes then elongated and penetrated the stigma. Hand-selfed flowers produced as many seeds as hand-crossed flowers. There was no significant difference in seed production between pollinator-excluded flowers and hand-selfed flowers. When emasculated flowers were exposed to pollinators, they produced significantly fewer seeds than intact flowers. Visits by effective pollinators were rare., Conclusions: This study describes a new form of delayed autonomous self-pollination. As the predominant mechanism of sexual reproduction in R. debilis, delayed self-pollination ensures reproduction when pollinators are scarce.

Objective: To investigate whether formalin inflammatory pain can induce the change in heme oxygenase-1 (HO-1) expression in the spinal cord of rats or not and the time course character., Methods: 42 SD rats were divided into 7 groups (n = 6): control formalin 6 h, formalin 12 h, formalin 1 d, formalin 2 d, formalin 3 d and formalin 7 d groups. Rats were subcutaneously injected with 0.2 ml 0.5% formalin into the ventral surface of right hind paw to induce periphery inflammatory pain. The immunohistochemistry was used to observe the expression of HO-1 protein in laminae I - II of the spinal cord dorsal horn and the area around canalis centralis of the I5 spinal segment of rats., Results: There are rare HO-1 immunoreactive cells in the laminae I - II of the dorsal horn and the area around canalis centralis of the I5 spinal segment of rats of control group and HO-1 immunoreactive cells were light in staining degree. Comparing with control group, the numbers of HO-1 immunoreactive cells in the I - II laminae of dorsal horn and area around canalis centralis were increased in the rats at 6 h after formalin injection. The number and staining degree of HO-1 immunoreactive cells were further increased at 12 h and peaked at 1 d after formalin injection. They didn't return to normal level at the 7th day. There were no difference in right and left dorsal horn in the number and staining degree of HO-1 immunoreactive cells at the same time after formalin injection., Conclusion: Formalin inflammatory pain induced increased expression of HO-1 in the spinal cord dorsal horn and the area around canalis centralis of rats. At 1 d after injection of formalin, the increased expression of HO-1 was the most obviously.

Objective: Division of the CA1, CA3 and dentate gyrus (DG) regions of the hippocampus of Wistar rat under the stereomicroscope., Methods: Twenty-four Wistar rats were randomly assigned to three groups. (1) The brain was sectioned coronally (n = 6). The sections were stained with thionin and the morphology of cells in each region of the hippocampus was observed under microscopy. (2) The hippocampus was dissected out and observed on the whole. Then, the CA1, CA3 and DG regions of the hippocampus were divided. Every region divided was sectioned, and the morphology of cells was observed. (3) Rats with brain ischemia or not were also decapitated and the HSP 70 expressions were observed in CA1, CA3 + DG regions by Western blot and immunohistochemical staining (n = 12)., Results: (1) The CA1, CA3 and DG regions of the hippocampus could be clearly observed in coronal section of the brain stained by thionin. (2) Under the stereomicroscope, the CA1 and DG regions of the hippocampus could be separated along the hippocampal fissure between them in ventral surface of the hippocampus. The CA3 and DG regions of the hippocampus could be separated along a fissure between them. The appearance of cells in the sections of the divided CA1, CA3 and DG specimens is consistent with that in the brain coronal sections, respectively. (3) The results of Western blot indicated that the HSP 70 expression of the brain ischemia group was up-regulated significantly in CA3 + DG regions compared with the sham group. However, HSP 70 expression has no significant changes in CA1 region. The above results were consistent with those of the immunohistochemical staining., Conclusion: The CA1, CA3 and DG regions of the hippocampus of Wistar rat could be divided under stereomicroscope, and the divided each region was sensible for detection of protein using Western blot.

Objective: To observe whether formalin inflammatory pain can induce neuron apoptosis in rats spinal cord or not and the effects of nitric oxide on the spontaneous pain reaction and neuron apoptosis in the spinal cord of rats with formalin inflammatory pain., Methods: Formalin-induced paw licking time was used to reflect the degree of spontaneous pain of rats, and the flow cytometry was used to detecte neuron apoptosis rate of spinal cord., Results: Compared with control group, the apoptosis ratio of spinal neuron was increased in the rats with formalin inflammatory pain, and peaked at 3d after formalin injection. Pre-intrathecal injection of NOS inhibitor L-NAME inhibited the nociceptive behavioural response in double phases induced by fonnrmalin injection and cut down the neuron apoptosis ratio of spinal cord of rats with formalin inflammatory pain. Nociceptive behavioural response and incraesed neuron apoptosis in the spinal cord were induced by intrathecal injection of L-Arg in normal rats., Conclusion: The results indicated that formalin inflammatory pain could induce the apoptosis of spinal neurons. The neurons apoptosis was the most significant on the third day after formalin injection. The increased pruduction of NO in spinal cord could promote the transmit of nociceptive information and participate the induction of neuronal apoptosis during the formalin inflammatory pain.

Objective: The PAX6 gene encodes a transcriptional regulator involved in oculogenesis and other developmental processes such as aniridia, a congenital condition characterized by the underdevelopment of the iris of eyes. The function of the PAX6 gene in these two conditions is still poorly defined. The purpose of this study is to identify the mutation of the PAX6 gene in a Chinese family with aniridia., Methods: Two aniridia patients collected from the family underwent full ophthalmologic examination. Genomic DNA was prepared from venous leukocytes of the two patients and five healthy individuals in the family, and 100 unrelated healthycontrols. Exons 4-13 and their immediate flanking sequences of the PAX6 gene was analyzed by PCR amplification, direct sequencing, and single-strand conformation polymorphism(SSCP)., Results: The sequencing result revealed a novel PAX6 mutation in the two patients. It was a heterozygous mutation (IVS10+1G>A) at the boundary of exon 10 and intron 10. The mutation was also detected by SSCP analysis. It was not detected in the healthy relatives and unrelated controls., Conclusion: Aniridia is an autosomal dominant inheritable disease. A novel PAX6 gene mutation has been identified in the Northeastern Chinese family with aniridia. The genetic analysis suggested that this novel mutation in the PAX6 gene is capable of causing the classic aniridia phenotype.

Objective: To better assess the role of p38 MAPK, this project was designed to investigate whether intraventricular injection of antisense oligodeoxynucleotide (As-ODN) directed against the p38 MAPK of pyramidal neurons in hippocampus could affect the brain ischemic tolerance induced by limb ischemic preconditioning (LIP)., Methods: The rat 4-vessel occlusion global cerebral ischemic model was used. Forty-eight male Wistar rats with permanently occlusion of the bilateral vertebral arteries were divided into 8 groups (n=6): sham, LIP, brain ischemic insult, LIP + brain ischemic insult, distilled water + LIP + brain ischemic insult, p38 MAPK As-ODN and p38 MAPK As-ODN + LIP + brain ischemic insult (two doses of 5 nmol/5 microl and 10 nmol/5 microl were used) groups. Thionin staining was used for observing histological changes of the hippocampus., Results: No significant delayed neuronal death (DND) was detected in the CA1 hippocampus of the rats that underwent sham and LIP operation. Brain ischemic insult for 8 min induced obvious DND as represented with the increase in histological grade (HG) and decrease in neuronal density (ND) significantly compared with sham and LIP groups. LIP protected the CA1 hippocampal pyramidal neurons against DND induced by global brain ischemic insult, suggesting the occurrence of brain ischemic tolerance. However, pretreatment with p38 MAPK As-ODN effectively blocked the ischemic tolerance induced by LIP in a dose dependent manner., Conclusion: It could be concluded that p38 MAPK plays an important role in the brain ischemic tolerance induced by LIP.

Aim: To study the effect of intrathecal injection of MK-801, a NMDA receptor antagonist, on the NOS activity and NO content of hippocampus in rat during the process of formalin-induced inflammatory pain as well as the pain behavior of rat., Methods: The degree of pain was determined by observing the time of licking and biting the injected paw. NOS expression in the hippocampus was determined by using NADPH-d histochemical staining. NO content of hippocampus was determined by assaying NO3; and NO2., Results: Subcutaneous injection of formalin elicited a characteristic pain behavioural response consisting of licking and biting the injected paw, etc. Intrathecal injection of MK-801 could shorten obviously the time of licking and biting representing pain behavioural response in phase 2. It is suggested that intrathecal injection of MK-801 could block the pain behavioural response induced by formalin (P < 0.05). The number and staining degree of NADPH-d positive neurons in formalin group significantly increased at 12 h after the formalin injection in CA1, CA2-3 and DG of hippocampus compared with control group as well as NO content, however, the number and staining degree of NADPH-d positive neurons in formalin + MK-801 group significantly decreased in contrast to those of formalin 12 h group as well as the NO content (P < 0.01)., Conclusion: Intrathecal injection of NMDA receptor antagonist MK-801 could inhibit the NOS activity and NO production in hippocampus of rat, which showed the increase of hippocampal NO production was mainly induced by the peripheral nociceptive information input.

Aim: To explore the role of nitric oxide (NO) resulted from nNOS in the mGluR2/3 mediated-brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP), the present study is undertaken to observe the influences of alpha-methyl-(4-tetrazolyl-phenyl) glycine (MTPG), an antagonist of mGluR2/3, on the expression of nNOS during the induction of the brain ischemic tolerance based on confirming the blocking effect of MTPG on the induction of the tolerance., Methods: Thirty-six Sprague-Dawley rats, whose vertebral arteries were permanently occluded, were randomly divided into sham, CIP, ischemic insult, CIP+ ischemic insult, MTPG+ CIP and MTPG+ CIP+ ischemic insult groups. Thionin staining and immunohistochemistry were used for neuropathological evaluation and assay of nNOS expression in the hippocampal CA1 subregion of the rats., Results: The expression of nNOS showed moderate and extreme up-regulation in the CIP and ischemia groups, respectively, compared to the sham group. The preceded CIP blocked in certain extent the extreme up-regulation of nNOS induced by brain ischemia in CIP + ischemia group. Administration of MTPG via lateral cerebral ventricle 20 min before CIP blocked the up-regulation of nNOS induced by CIP, but had no influence on the pyramidal neuronal survival. While in the MTPG+ CIP+ ischemic insult group, the expression of nNOS was stronger than that in the MTPG + CIP group, and the up-regulation was accompanied with obvious delayed neuronal death. Discussion concerned illustrated that the relative intensive up-regulation of nNOS in this group might be attributed to brain ischemia other than MTPG., Conclusion: NO resulted from nNOS participated the induction of mGluR2/3 mediated-brain ischemic tolerance as a downstream molecule of activation of mGluR2/3 during CIP.

Aim: To investigate whether formalin inflammatory pain can induce hippocampal neuronal apoptosis of rats or not., Methods: Rats were subcutaneously injected with 0.2 ml 0.5% formalin into the ventral surface of right hind paw to induce periphery inflammatory pain. The flinches of rats were counted to observe their painful reaction. Flow cytometry was used to assay the ratio of apoptosis of hippocampal neurons. The immunohistochemistry was used to observe the expression of p53 protein in hippocampal subregions., Results: Compared with control group, the apoptotic ratio of hippocampal neurons was significantly increased in rats with inflammatory pain, and formalin inflammatory pain induced upregulation of p53 protein expression in all hippocampal subregions. Both the apoptotic ratio and the p53 protein expression peaked on the third day after the formalin injection. The twice injection of formalin into the hind paws of rats resulted in an enhancement of painful reaction and increase in apoptotic ratio of hippocampal neurons compared with the rats of injection formalin once group., Conclusion: Formalin inflammatory pain can induce the hippocampal neuronal apoptosis in rats with a certain time course. Neuronal apoptosis is relevant to the intensity of pain. The up-regulation of p53 protein expression may implicate in the induction of hippocampal neuronal apoptosis in rats with inflammatory pain.

Aim: To further explore the role of adenosine A1 receptor in the neuroprotective effect of cerebral ischemic preconditioning, the present study was undertaken to observe the effect of inhibiting expression of adenosine Al receptor with adenosine A1 receptor antisense oligodeoxynucleotide (ARA1 As-ODN) on the neuroprotective effect of cerebral ischemic preconditioning against delayed neuronal death (DND) normally induced by lethal brain ischemia., Method: The rat 4-vessel occlusion global cerebral ischemic model was used. Forty-eight male Wistar rats with permanent occlusion of the bilateral vertebral arteries were divided into 8 groups: Sham, CIP, brain ischemic insult, CIP + brain ischemic insult, Distilled water + CIP + brain ischemic insult, ARA1 As-ODN, ARA1 As-ODN +CIP, ARA1 As-ODN+ CIP + brain ischemic insult(two doses of 10 nmol/5 microl and 20 nmol/5 microl were used) groups. ARA1 As-ODN was dissolved in distilled water and injected into the right lateral cerebral ventricle. To illustrate the profile of DND, histological grade (HG) and neuronal density (ND) in the CA1 region of the hippocampus were examined 7 d after the sham operation or the last time of ischemia under thionin staining., Results: The HG and ND in CIP group were similar to those in sham group. Brain ischemic insult induced obvious DND as represented with the increase in HG and decrease in ND significantly (P < 0.05 vs. sham and CIP groups). In CIP + ischemic insult group,no obvious DND was observed,which indicated that CIP protected pyramidal neurons against the ischemic insult.While the administration of ARA1 As-ODN in ARA1 As-ODN + CIP + brain ischemic insult group caused obvious increase in HG and decrease in ND compared with CIP + brain ischemic insult group (P < 0.05) in a dose dependent manner,which indicated that the neuroprotective effect of CIP against DND of hippocampal pyramidal neurons normally induced by ischemic insult was inhibited by the administration of ARA1 As-ODN., Conclusion: The results further demonstrate the association of up-regulation of adenosine A1 receptors with the induction of CIP-mediated BIT.

Background and Aims: Reproductive assurance, the ability to produce seeds when pollinators or mates are scarce, is thought to be the major advantage of selfing in flowering plants. However, few studies have performed a direct cost-benefit analysis of the selective advantage of selfing, particularly given a long-term perspective among populations or across several flowering seasons within population. This study examined the fertility consequences of autonomous selfing in Roscoea schneideriana (Zingiberaceae), a small perennial Himalayan ginger typically found in habitats at around 3000 m a.s.l., Methods: The floral biology of R. schneideriana was studied in natural populations; the capacity for autonomous selfing was estimated using pollinator exclusion experiments; the timing of selfing was quantified by anther removal at different times during flowering; whether autonomous selfing increases seed production was tested by emasculating flowers; and the magnitude of inbreeding depression was estimated by comparing relative performance of progeny from self- and cross-pollinations. Pollinator observations were also conducted in the natural populations., Key Results: The hooked stigmas of most flowers curl towards the anther and can contact pollen grains at an early stage of anthesis. Flowers with potential pollinators excluded set of as many seeds per fruit as hand-selfed and opened flowers. Autonomous selfing mostly occurs within 2 d of anthesis and can increase seed production by an average of 84 % in four populations during the flowering seasons of 2005-2007. Visits by effective pollinators were extremely rare. The cumulative inbreeding depression of R. schneideriana was 0.226., Conclusions: Autonomous selfing in R. schneideriana is achieved by stigmas curling towards the anthers early in flowering. It is suggested that under the poor pollination conditions, autonomous selfing has been selected for in this alpine ginger because it provides substantial reproductive assurance with very low costs.

The present study was undertaken to investigate the role of glial glutamate transporter-1 (GLT-1) in the brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP) by observing the effect of GLT-1 antisense oligodeoxynucleotides (AS-ODNs) on the neuro-protection of CIP against brain ischemic insult in rats. Wistar rats with permanently occluded bilateral vertebral arteries were randomly assigned to 7 groups: (1) Sham group: the bilateral common carotid arteries (BCCA) were separated, but without occluding the blood flow; (2) CIP group: the BCCA were clamped for 3 min; (3) Brain ischemic insult group: the BCCA were clamped for 8 min; (4) CIP+brain ischemic insult group: 3 min CIP was preformed 2 d prior to 8 min ischemic insult; (5) Double distilled water group: 5 muL double distilled water was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively; (6) AS-ODNs group: 5 microL AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively. This group was further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs; (7) AS-ODNs+CIP+brain ischemic insult group: 5 microL AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after CIP, respectively. This group was also further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs. The other treatments were the same as those in CIP+brain ischemic insult group. The effect of the AS-ODNs on the expression of GLT-1 was assayed by using Western blot analysis. The profile of delayed neuronal death (DND) of pyramidal neurons in the CA1 hippocampus was evaluated by using thionin staining under light microscope by determining the neuronal density (ND) and histological grade (HG). Western blot analysis showed that AS-ODNs injected into the lateral cerebroventricle inhibited the expression of GLT-1 in the CA1 hippocampus in a dose-dependent manner. Neuropathological evaluation showed that there was no apparent DND in sham and CIP groups. Obvious DND of pyramidal neurons was found in brain ischemic insult group, which was represented by an increase in HG and a decrease in ND. CIP effectively protected the pyramidal neurons in the CA1 hippocampus against DND normally induced by ischemic insult, which indicating that CIP induced ischemic tolerance on the pyramidal neurons in the CA1 hippocampus. However, the injection of AS-ODNs into the lateral cerebroventricle blocked the neuro-protection of CIP against DND induced by brain ischemic insult. These results further proved the role of GLT-1 in the brain ischemic tolerance induced by CIP in rats.

Background and Aims: Flexistyly is a sexual dimorphism where there are two morphs that differ in the temporal expression of sexual function and also involve reciprocal movement of the stigmatic surface through a vertical axis during the flowering period. The adaptive significance of flexistyly has been interpreted as a floral mechanism for outcrossing, but it may also function to reduce sexual interference in which styles and stigmas impede the pollen export. Here these two explanations of flexistyly were tested in Alpinia blepharocalyx through a hand-pollination experiment., Methods: Hand-pollinations were performed in two temporal morphs and consisted of two sequential pollination treatments, namely self-pollination in the morning and inter-morph pollination in the afternoon (treatment 1) or conversely inter-morph pollination in the morning and self-pollination in the afternoon (treatment 2), and two simultaneous self- and inter-morph cross-pollination treatments either in the morning (treatment 3) or in the afternoon (treatment 4). Seed paternity was then determined to assess relative success of self- versus cross-pollen using allozyme markers., Key Results: In the sequential pollination treatments, whether the stigmas of recipients are receptive in the morning is crucial to the success of the pollen deposited. When the cataflexistylous (protandrous) morph served as pollen recipient, early-arriving pollen in the morning can sire only a very small proportion (<15%) of seeds because the stigmas were then unreceptive. However, when the anaflexistylous (protogynous) morph served as pollen recipient, early pollen did gain a large competitive advantage over the late pollen, particularly when cross-pollen arrived first. Simultaneous self- and inter-morph cross-pollination indicated that outcross-pollen is more competitive than self-pollen on receptive stigmas., Conclusions: Differential maturing of male and female organs in Alpinia blepharocalyx is sufficient for selfing avoidance, obviating the need for style movements. Instead, the upward style curvature of the cataflexistylous morph in the morning and the anaflexistylous morph in the afternoon most likely represents a means of reducing interference with pollen export.

Aim: To observe the changes of nitric oxide synthase (NOS) activity and nitric oxide (NO) content of hippocampus including their time course and region distribution character in rat during the process of formalin-induced inflammatory pain as well as the pain behavior of rat., Methods: The pain threshold (PT) was determined by radiant heat-induced tail flick test. NOS expression in the hippocampus was determined by using NADPH-d histochemical staining. NO production in hippocampus was determined by assaying NO3- and NO2-., Results: Subcutaneous injection of formalin elicited nociceptive behavioural response and led to decrease in PT of rat. The number and staining degree of NADPH-d positive neurons began to increase at 6 h after the formalin injection in CA1, CA2 - 3 and DG of hippocampus as well as NO content, which increased most obviously at 12 h and returned to control level at 48 h., Conclusion: Formalin-induced inflammatory pain could induce the elevation of NOS activity in CA1, CA2 - 3 and DG of hippocampus with a certain time course, which further led to a increase of NO production in hippocampus.

Aim: To explore the role of femoral nerves section (FNS) on the protection of limb ischemic preconditioning (LIP) against cerebral ischemia/reperfusion injuries., Methods: Model of brain ischemia induced by Four-vessel occlusion was used. LIP was performed by clamping the bilateral femoral arteries for 10 min 3 times in a interval of 10 min. Rats with vertebral arteries permanently occluded were divided into sham group, cerebral ischemic group, FNS + cerebral ischemic group, LIP + cerebral ischemic group, FNS + LIP + cerebral ischemic group. The changes of neural density (ND) in the CA1 hippocampus were observed 7d after the sham operation or brain ischemia under thionin staining. The expression of c-Fos in the CA1 hippocampus was measured 6 h after the sham operation or brain ischemia under immunohistochemistry method., Results: Thionin staining revealed that serious neuronal damage was visualized in the CA1 hippocampus in both cerebral ischemic group and FNS + cerebral ischemic group as compared with sham group. LIP attenuated the neuronal damage of the CA1 subfield induced normally by cerebral ischemia/reperfusion, and ND in LIP + cerebral ischemic group was significantly higher than that in cerebral ischemic group (P < 0.01). But obvious neuronal damage of the CA1 subfield was found in FNS+ LIP + cerebral ischemic group, and ND was significantly decreased as compared with LIP + cerebral ischemic group (P < 0.01). These results suggested that the protection of LIP against cerebral ischemia/reperfusion injuries might be cancelled by preceding section of femoral nerve. It was found that there was almost no c-Fos expression in the CA1 hippocampus in sham group. Changes of c-Fos expression in the CA1 subfield in cerebral ischemic group were similar to that in sham group. But in LIP + cerebral ischemic group, c-Fos expression in the CA1 subfield was markedly increased and the number of positive cells and optical density of c-Fos expression were significantly higher than those in sham and cerebral ischemic group. c-Fos expression in the CA1 subfield was again decreased in FNS + LIP + cerebral ischemic group, and the number of positive cells and optical density of c-Fos expression were significantly lower than those in LIP + cerebral ischemic group., Conclusion: Neural pathway participated in the protective effect of LIP on brain, and increased c-Fos expression in the CA1 hippocampus by LIP after cerebral ischemia/reperfusion, might be a part of neural pathway by which LIP induced brain ischemic tolerance.

Aim: To study the effect of different intervals between occlusions of vertebral arteries and bilateral common carotid arteries on the Pulsinelli 4-vessel occlusion global cerebral ischemic model, and the features of ischemia of the brainstem and hippocampus induced by occulusion of bilateral common carotid arteries under the condition of occlusion of unilateral vertebral artery., Methods: Eighty four adult male Wistar rats were divided into 4 groups randomly: control group, bilateral vertebral artery occluding group, global brain ischemic insult group, and unilateral vertebral artery occluding plus bilateral common carotid arteries occluding group. In the global brain ischemic insult group, rats were further divided into 24 h, 48 h, and 72 h interval subgroups according to the interval between the occlusion of the vertebral arteries and bilateral common carotid arteries. The responses including enlarging of pupils and the light reflex during the brain ischemia were observed. The duration of right reflex disappearing, the general state, and the delayed neuronal death (DND) of pyramidal neurons in the CA1 hippocampus of the rats after the brain ischemia were also observed., Results: Among the global brain ischemic insult group, both the responses and DND were more severe in 72 h interval subgroup than those in 24 h and 48 h interval subgroups. There was no significant difference between 24 h and 48 h interval subgroups. When the bilateral common carotid arteries were occluded under the condition of occlusion of unilateral vertebral artery, severe DND was observed in the CA1 hippocampus ipsilateral to the occluding vertebral artery, but no significant DND was observed in the contralateral CA1 hippocampus., Conclusion: The results suggested that the prior occlusion of the bilateral vertebral arteries during producing Pulsinelli 4-vessel occlusion global cerebral ischemic model might be a cerebral ischemic preconditioning that could protect to some extent pyramidal neurons of the hippocampus against severe ischemic insult induced by occlusion of bilateral common carotid arteries within 48 h. Moreover, There is ipsilateral predominance of blood perfusion from one side of vertebral artery to the brainstem and hippocampus, although there was Willis artery circle in rats.

Background and Aims: Despite considerable investment in elaborate floral displays, Tacca chantrieri populations are predominantly selfing. It is hypothesized that this species might possess considerable spatial or temporal variation in outcrossing rates among populations. To test this hypothesis, genetic variability and genetic differentiation within and among T. chantrieri populations were investigated to find out if they are in agreement with expectations based on a predominantly inbred mating system., Methods: Genetic diversity was quantified using inter-simple sequence repeats (ISSR) in 303 individuals from 13 populations taken from known locations of T. chantrieri in China, and from one population in Thailand., Key Results: Of the 113 primers screened, 24 produced highly reproducible ISSR bands. Using these primers, 160 discernible DNA fragments were generated, of which 145 (90.62 %) were polymorphic. This indicated considerable genetic variation at the species level. However, there were relatively low levels of polymorphism at population levels, with percentages of polymorphic bands (PPB) ranging from 8.75 % to 55 %. A high level of genetic differentiation among populations was detected based on different measures (Nei's genetic diversity analysis: G(ST) = 0.5835; AMOVA analysis: F(ST) = 0.6989). Furthermore, based on levels of genetic differentiation, the 14 populations clustered into two distinct groups separated by the Tanaka Line., Conclusions: High levels of differentiation among populations and low levels of diversity within populations at large spatial scales are consistent with earlier small-scale studies of mating patterns detected by allozymes which showed that T. chantrieri populations are predominantly selfing. However, it appears that T. chantrieri has a mixed-mating system in which self-fertilization predominates, but there is occasional outcrossing. Significant genetic differences between the two distinct regions might be attributed to vicariance along the Tanaka Line. Finally, possible mechanisms of geographic patterns based on genetic differentiation of T. chantrieri are discussed.