Polyploidy Induction and Morphological Characteristics in the Asiatic Hybrid Lily(Lilium spp.cultivar dazzling)

The Asiatic hybrid lily cultivar Lilium dazzling(family Liliaceae)is a perennial herbaceous bulbous plant with bright yellow flowers of ornamental values.It also shows resistance to diseases and abiotic stress,making it an ideal parent for breeding studies.This study established a sterile culture system using scales of dazzling lily as explants to induce polyploidy.Adventitious buds growing to 1 cm were treated with different concentrations of colchicine(125,250 and 500μmol·L-1),pendimethalin or trifluralin(100,200 and 300μmol·L-1,respectively)for 12,24 and 36 h.Flow cytometry and chromosome counting were used to identify ploidy,and the phenotype of the polyploids was examined by stomatal observation and leaf index determination.With increasing mutagen concentration and exposure time,the mortality and mutagenic rate increased.The optimal treatment group was:300μmol·L-1 trifluralin for 12 h,which resulted in 15.55%mortality rate and 42.22%induction rate.The polyploid plants showed a significant increase in stomata length,leaf length and leaf width,with a decrease in stomatal density compared with the control plants.This study provided a basis for polyploid breeding.

Habitat Suitability and Distribution Pattern Response to Global Climate Change in a Widespread Species,the Asiatic Toad(Bufo gargarizans)

The distribution and diversity of the species are closely related to the global climate.As the most widely distributed species of Bufonidae in China,the study of the distribution pattern and habitat suitability of the Asiatic toad under climate change can help us understand the reply pattern of Bufonidae habitat to climate change.Here,combined with the Maxent model and GIS technology,the effects of climate change on the distribution pattern and habitat suitability of the Asiatic toad were comprehensively analyzed.The results show that the rainfall during the wettest season(Bio16)and the mean temperature of the driest season(Bio9)have a considerable impact on the distribution of the Asiatic toad.In the next 30 to 50 years,across the overall spacial scale of the Chinese mainland,the habitat of the Asiatic toad will be primarily in the eastern part of China and less in south part,while its distribution area will expand to the midwest and northwest parts of China.Overall,the area in which it can be distributed will be reduced and suitable habitat will shift to some regions of higher latitude and elevation.In a word,we systematically analyzed the changes of the distribution pattern and habitat suitability of the Asiatic toad with climate change,and we aim to provide data on how climatic variation may impact amphibians.

Analysis of Protein and Isoenzyme of LiLum davidii var.unicolor,Lilium Asiatic Hybrids and Their Hybrid F_1

[Objective]The paper was to provide theoretical basis for selection of parental combination and early identification of hybrids.[Method]The soluble protein and peroxidase of LiLum davidii var.unicolor,Lilium Asiatic hybrids and their filial generations were analyzed using polyacrylamide gel electrophoresis technique.[Result]The protein spectrum of filial generation with L.davidii var.unicolor as parent not only appeared the homologous band as parent with darker coloring,but also had new bands compared with parent.Peroxidase zymogram of hybrid F1 mainly displayed incomplete complementary and hybrid type of parent.[Conclusion]Protein spectrum and peroxidase zymogram could be used as the biochemical markers for the identification of hybrids of lily,which could also detect the target traits of plant.

The anti-inflammatory effects of asiatic acid in lipopolysaccharide-stimulated human corneal epithelial cells

AIM： To investigate the anti-inflammatory effects of asiatic acid（AA） on lipopolysaccharide（LPS）-induced inflammatory response in human corneal epithelial cells（HCECs）.METHODS： Cell viability was measured using a cell counting kit-8（CCK-8） assay.Quantitative real-time polymerase chain reaction（qR T-PCR） was used to determine the mR NA expression of interleukin-8（IL-8）,interleukin-6（IL-6）,interleukin-1β（IL-1β）,tumor necrosis factor-alpha（TNF-α）,and transforming growth factor-β（TGF-β） in HCECs.Intracellular reactive oxygen species（ROS） was measured using the ROS assay kit.Glutathione（GSH） concentration was measured using the total GSH assay kit.Akt1 and Akt phosphorylation（p-Akt1） levels were measured by Western blotting and immunofluorescence.RESULTS： AA induced toxicity at high concentrations and significantly stimulated the proliferation of HCECs at concentrations of 20 μmol/L for 1h.LPS at concentrations of 300 ng/mL for 1h significantly stimulated the mR NA expression of IL-8,IL-6,IL-1β,TNF-α,and TGF-β in HCECs,while the stimulation effects were significantly inhibited by AA（20 μmol/L）.In addition,AA was found to decrease the content of ROS,increase GSH generation,and also inhibit LPS-induced p-Akt in HCECs.CONCLUSION： AA decreases the generation of inflammatory factors IL-8,IL-6,IL-1β,TNF-α,and TGF-β in LPSstimulated HCECs.AA significantly inhibites the intracellular concentrations of ROS and increases GSH generation.AA also inhibites LPS-induced p-Akt in HCECs.These findings reveal that AA has anti-inflammation effects in LPS-stimulated HCECs.

Status of Asiatic Wild Cat and its habitat in Xinjiang Tarim Basin, China

The Asiatic Wild CatFelis silvestris ornataisregarded as “Least concerned” (LC) first, as “Vulnerable” (VU) and following “Endangered” (EN) and then “Critically endangered” (CR) species as finally and originally concentrative distribute in Xinjiang Tarim Basin region in northwest China. This paper provides comprehensive information on bio-morphology, habitat selectivity, environmental condition, habit, preyfeed source and every item of composition, and relationship among Wild Cat and domestic cat at presence investigated in 2004-2006 and 2011- 2013 as especial study. The paper also illustrated some dynamical statistics of wild cat’ pelt collection by national trade from three prefectures in the last 40 years. Briefing the results of indication that large scale and continuous openup land for cotton in unplanned, exploit petroleum and natural gas, misapply water and destroy desert vegetation, poaching and killing prey objectives, etc., of them long-term effects on plants functional density and qualities of the cat surviving habitat and productivity, whereas pressures of increasing human population to fragile desert ecosystem showed efficiency and desirable effects of the cat surviving.

Molecular cloning, characterization, and antioxidant function of catalase in Lymantria dispar asiatic (Lepidoptera: Lymantriidae) under avermectin stress

The critical antioxidant catalase(CAT)breaks down hydrogen peroxide induced by environmental stresses.Here we cloned full length catalase cDNA from Lymantria dispar asiatic(LdCAT).Bioinformatic analyses showed that open reading frames of LdCAT contains 1524 bp,encoding 507 amino acids with molecular weight of 126.99 kDa,theoretical pI of 5.00,aliphatic index of 29.92,grand average of hydropathicity of 0.764,and instability index(II)of 46.56.Protein BLAST and multiple sequence alignment indicated that LdCAT had high identity with CAT from other insects,especially lepidopterans.In a phylogenetic analysis,LdCAT was most similar to CAT from Spodoptera litura and S.exigua.Quantitative realtime polymerase chain reaction showed that LdCAT transcripts in all instar larvae and the five tissues tested,verifying the ubiquity of LdCAT in L.disapr.Moreover,LdCAT of third instar larvae was significantly upregulated after they fed on avermectin at sublethal and LC10 doses.The highest relative transcript levels were found 2 h after an avermectin spray at LC90,and in the cuticula,rather than heads,fat bodies,malpighian tubes,and midguts after a spray avermectin at a sublethal concentration.The expression level of LdCAT under pesticide stresses here suggested that CAT is an important antioxidant enzyme of L.disapr defensing against pesticide stress and may be a good target for controlling this pest.

Asiatic acid-pectin hydrogel matrix patch transdermal delivery system influences parasitaemia suppression and inflammation reduction in P. berghei murine malaria infected Sprague-Dawley rats

Objective: To report the influence of transdermal delivery of asiatic acid(AA) in Plasmodium berghei-infected Sprague Dawley rats on physicochemical changes, %parasitaemia and associated pathophysiology. Methods: A topical once-off AA(5, 10, and 20 mg/kg)- or chloroquine(CHQ)-pectin patch was applied on the shaven dorsal neck region of Plasmodium berghei-infected Sprague Dawley rats(90-120 g) on day 7 after infection. Eating and drinking habits, weight changes, malaria effects and %parasitaemia were compared among animal groups over 21 d. Results: AA-pectin patch application preserved food and water intake together with %weight gain. All animals developed stable parasitaemia(15%-20%) by day 7. AA doses suppressed parasitaemia significantly. AA 5 mg/kg patch was most effective. AA and CHQ displayed bimodal time-spaced peaks. CHQ patch had a longer time course to clear parasitaemia. Conclusions: AA influences bio-physicochemical changes and parasitaemia suppression in dose dependent manner. In comparison by dose administered, AA has much better efficacy than CHQ. AA may be a useful antimalarial. AA and CHQ displays bimodal peaks suggesting possible synergism if used in combination therapy.

The recovery and protective effects of asiatic acid on differentiated human neuroblastoma SH-SY5Y cells cytotoxic-induced by cholesterol

Objective:To investigate the effect of asiatic acid(AA) on the differentiated human neuroblastoma SH-SY5 Y cells cytotoxic-induced by cholesterol.Methods:Human neuroblastoma SH-SY5 Y cells were either exposed to different concentrations of AA or treated with different doses of cholesterol to reveal their responding viability by MTT assay.The selective 1 mmol/L concentration of AA was then used to test for either the protective or the recovery effects on the cells treated with 250 mmol/L concentration of cholesterol.Results:AA has a propensity to directly increase the viability of differentiated human neuroblastoma SH-SY5 Y cells.Cholesterol has significant cytotoxic effect on those cells in a concentration-dependent manner.AA has the ability to slightly recover the viability of the differentiated culture cytotoxic-induced by cholesterol but could not protect those cells from cytotoxic-induced by cholesterol.Conclusions:High concentrations of cholesterol were observed to be harmful to the neurons and AA had a slight effect of reducing neuronal death caused by cholesterol.

Blood Typing of Asiatic Black Bear (Ursus thibetanus)

[Objective]The paper was to analyze whether Asiatic black bear( Ursus thibetanus) had different blood group systems. [Method]Whole blood samples were collected from 40 Asiatic black bears in Fujian Province,China. Tube method was used to detect antibodies in plasma,and antibody isotype was determined with 2-mercaptoethanol. Plasma was further analyzed by mass spectrometry. [Result] The plasma from four black bears had antibodies,possibly Ig M isoform,which could agglutinate RBCs from 30 bears. Blood samples from 10 bears were tested by human blood typing reagents. The results showed that four black bears had blood type like human type O,while six bears had like human type B. Mass spectrometry results demonstrated that plasma protein had extensive homology to serum albumin-like isoform 1 found in giant panda( Ailuropoda melanoleuca). [Conclusion]Asiatic black bear might have at least one blood group system with two blood types. If the sick bear needs blood transfusion,a cross-matching test is necessary. Moreover,giant panda might receive blood from Asiatic black bear in case of emergency.

Asiatic acid inhibits lung cancer cell growth in vitro and in vivo by destroying mitochondria

Asiatic acid(AA), a pentacyclic triterpene found in Centella asiatica, displays significant antiproliferative effects on cancer cells in vitro although the underlying mechanism of this effect remains unknown. This study investigated the efficacy and mechanism of action of AA against lung cancer both in vivo and in vitro. Using the MTT assay, AA was found to induce apoptosis in a dose- and timedependent manner, an effect enhanced by pretreatment with an autophagy inhibitor. It also elevated expression of microtubule-associated protein 1 light chain 3(LC3) and decreased the expression of p62.Furthermore, exposure to AA resulted in collapse of the mitochondrial membrane potential and generation of reactive oxygen species(ROS), suggesting mitochondria are the target of AA. In the mouse lung cancer xenograft model, oral administration of AA significantly inhibited tumor volume and weight accompanied by significant apoptosis of lung cancer cells. In addition, it led to a significant decrease in the expression of proliferating cell nuclear antigen(PCNA). In summary, the results show that AA significantly reduces lung cancer cell growth both in vitro and in vivo and that the associated apoptosis is mediated through mitochondrial damage.

Asiatic acid mitigates hyperglycemia and reduces islet fibrosis in Goto-Kakizaki rat,a spontaneous type 2 diabetic animal model

The Goto-Kakizaki(GK) rat is a spontaneous type 2 diabetic animal model,which is characterized by a progressive loss of beta islet cells with fibrosis.In the present study,the hypoglycemic effect of asiatic acid(AA) in GK rats was examined.GK rats receiving AA at a daily dose of 25 mg·kg-1 for four weeks showed a significant reduction in blood glucose levels.Age-matched normal Wistar rats were given 0.5% sodium carboxymethyl cellulose(CMC-Na) solution for the same periods and used as control.Compared to the normal Wistar rats,GK rats treated with AA showed improvement in insulin resistance partially through decreasing glucose level(P < 0.01) and insulin level(P < 0.05).Furthermore,the results of immunohistochemistry indicate that AA treatment reduced islet fibrosis in GK rats.Fibronectin,a key protein related to islet fibrosis,was over-expressed in GK rats,which was reversed significantly by AA treatment(P < 0.05).These findings suggest that AA has a beneficial effect on lowering blood glucose levels in GK rats and improves fibrosis of islets in diabetes,which may play a role in the prevention of islets

Metal accumulation in Asiatic clam from the Lower Min River (China) and implications for human health

Considering growing concerns regarding polluted estuaries and their adverse effects on public health,this study aimed to identify concentrations of metal (Zn,Fe,Cr,Ni,Cd,Mn,As,Cu,and Pb) in Asiatic clams sampled along the Lower Min River,China.Multivariate methods were used to identify and apportion pollution sources.Noncarcinogenic and carcinogenic health risk assessments were performed to gauge adverse consumer health effects.Results showed that Cr,Pb,and Zn concentrations were higher than the limits prescribed in Chinese government guidelines.In comparison with concentrations of selected metals in other rivers,Cr,Pb,Zn,and As concentrations in clams were generally higher.Pollution assessment using the metal pollution index showed that sampling sites surrounding developing industrial and residential areas were the most polluted.Principal component analysis indicated significant anthropogenic metal contributions in clams.Health risk assessment indicated significant risk for clam consumers along the Lower Min River in terms of hazard quotient and carcinogenic risk and,thus,clam consumption from the study area should be avoided.The present findings would help in establishing environmental monitoring plans and contribute to preserving public health as well as the development of water conservation strategies to alleviate the metal pollution.

Hibernation with rhythmicity:the circadian clock and hormonal adaptations of the hibernating Asiatic toads(Bufo gargarizans)

Hibernation is one of the fundamental strategies in response to cold environmental temperatures.During hibernation,the endocrine and circadian systems ensure minimal expenditure of energy for survival.The circadian rhythms of key hormones,melatonin(MT),corticosterone(CORT),triiodothyronine(T3),and thyroxine(T4),and the underlying molecular regulatory mechanisms of hibernation have been well determined in mammals but not in ectotherms.Here,a terrestrial hibernating species,Asiatic toad(Bufo gargarizans),was employed to investigate the plasma CORT,MT,T3,and T4;and the retina,brain,and liver mRNA expression of the core clock genes,including circadian locomotor output cycles kaput(Clock),brain and muscle ARNT-like 1(Bmal1),cryptochrome(Cry)1 and 2,and period(Per)1 and 2,at 7-time points over a 24-h period under acute cold(1 day at 4℃),and hibernation(45 days at 4℃).Our results showed that the circadian rhythms of the core clock genes were rather unaffected by acute cold exposure in the retina,unlike the brain and liver.In contrast,during hibernation,the liver clock genes displayed significant circadian oscillations,while those in the retina and brain stopped ticking.Furthermore,plasma CORT expressed circadian oscillations in both groups,and T3 in acute cold exposure group,whereas T4 and MT did not.Our results reveal that the plasma CORT and the liver sustain rhythmicity when the brain was not,indicating that the liver clock along with the adrenal clock synergistically maintains the metabolic requirements to ensure basic survival in hibernating Asiatic toads.

Spectroscopic Studies on the Interaction of Asiatic Acid with Bovine Serum Albumin

Fluorescence spectroscopy, Fourier transform infrared （FT-IR） spectroscopy, circular dichroism （CD） and FT-Raman spectroscopy were employed to analyze the binding of the asiatic acid （AA） to bovine serum albumin （BSA） under simulative physiological conditions. Fluorescence data revealed that the fluorescence quenching of BSA by AA was the result of the formation of BSA-AA complex. The fluorescence quenching mechanism of BSA by AA was a static quenching procedure. According to the Van＇t Hoff equation, the thermodynamic parameters enthalpy change （△H0） and entropy change （△S0） for the reaction were evaluated to be --12.55 kJ·mol^-1 and 67.08 kJomol 1, respectively, indicating that hydrophobic and electrostatic interactions played a major role in stabilizing the complex. The influence of AA on the conformation of BSA has also been analyzed on the basis of FT-IR, CD and FT-Raman spectra.

The Asiatic Mode of Production in World History Perspective: From a Universal to a Particularistic View of History

Discussions on Marx＇s ＂Asiatic mode of production＂ are essentially about the relationship between the universal and the particularistic view of history. Marx adhered to the world history position, but in the course of formulating his theory of the Asiatic mode of production he moved from universal history to a particularistic view of history; whereas the shift from the ＂Russian path＂ and ＂Russian way＂ provided by the Russian revolution to Stalin＇s doctrine of the ＂five forms＂ of social development represented a return from particularistic to universal history. In developing a Chinese view of history, three elements cannot be overlooked： the Chinese pattern of historical development and China＇s present development experience; Marx＇s theory of social forms and his thoughts on Oriental society in his later years; and the ＂Russian path＂ and ＂systemic transformation＂ launched by the October Revolution.

Patterns of human-wildlife conflict and compensation practices around Daxueshan Nature Reserve, China

Understanding the spatial patterns of human-wildlife conflict is essential to inform management decisions to encourage coexistence, but it is constrained by the lack of spatially-explicit data. We collected spatially-implicit data of human-wildlife conflicts from 2009-2015 around Daxueshan Nature Reserve, Yunnan, China, and investigated the patterns and drivers of these conflicts. A questionnaire was also designed to capture local resident attitudes toward insurance-based compensation for the losses caused by targeted wildlife. We found that the Asiatic black bear （Ursus thibetanus） was the most conflict-prone animal around the reserve, followed by the rhesus macaque （Macaca mulatta） and Southeast Asian sambar （Cervus equinus）. Conflicts were unevenly distributed among seasons, villages, and communities, with several grids identified as conflict hotspots. Poisson models revealed that human-bear conflicts were negatively related to distance to the reserve and proportion of forest, but positively correlated to the proportion of cropland. Binomial models showed that communities affected by crop depredation were positively correlated with the proportion of cropland and negatively correlated with distance to the reserve, whereas communities affected by livestock depredation were negatively correlated with the proportion of cropland. The insurance-based scheme has compensated over 90% of losses, to the satisfaction of 90.6% of respondents. Our results suggest that human-bear conflict could be potentially reduced by eliminating food crops near the reserve boundary and livestock grazing at conflict hotspots.In addition, the insurance-based scheme could be replicated at a broader scale with improvement in loss assessment.

Repellent effect of guava leaf volatiles on settlement of adults of citrus psylla, Diaphorina citri Kuwayama, on citrus

The Asiatic citrus psyllid （Diaphorina cirri Kuwayama [Hemiptera： Sternorrhyncha： Psyllidae] is a vector ofhuanglongbing （citrus greening）, a devastating disease of citrus caused by phloem-limited bacteria. Growing guava （Psidium guajava） as an intercrop appears to be a successful means of reducing psyllid numbers within citrus orchards; however, the mechanism by which such a reduction is achieved is unknown. To determine the repellent effect of guava leaf and factors attributed to this activity, responses of adult psyllids to guava leaf and its odor were evaluated in cage tests and Y-tube olfactometer test. The results showed that guava leaf possessed a repellent effect against the adult citrus psyllids. Fewer psyllids were found on citrus leaves in the presence of guava foliage than in its absence. Young and old guava leaf showed equal repellent activity. By covering the guava shoots with net cloth, it was revealed that the repellent effect of guava leaf against adult psyllids on citrus was attributed to the volatile compounds, rather than physical factors. The olfactometer response of adult psyllids to guava leaf odor was dosage-dependent. Between guava odor and control, only 35.00%, 25.00% and 16.25% of the psyllids moved toward guava odor when presented with 5.0, 10.0 and 15.0 g of guava shoots, respectively. The olfactometer experiments also showed that both male and female psyllids responded similarly to the guava leaf odor.

Chinese herb microneedle patch for wound healing

Traditional Chinese medicine and Chinese herbs have a demonstrated value for disease therapy and sub-health improvement.Attempts in this area tend to develop new forms to make their applications more convenient and wider.Here,we propose a novel Chinese herb microneedle(CHMN)patch by integrating the herbal extracts,Premna microphylla and Centella asiatica,with microstructure of microneedle for wound healing.Such path is composed of sap extracted from the herbal leaves via traditional kneading method and solidified by plant ash derived from the brine induced process of tofu in a well-designed mold.Because the leaves of the Premna microphylla are rich in pectin and various amino acids,the CHMN could be imparted with medicinal efficacy of heat clearing,detoxicating,detumescence and hemostatic.Besides,with the excellent pharmaceutical activity of Asiatic acid extracted from Centella asiatica,the CHMN is potential in promoting relevant growth factor genes expression in fibroblasts and showing excellent performance in anti-oxidant,anti-inflammatory and anti-bacterial activity.Taking advantages of these pure herbal compositions,we have demonstrated that the derived CHMN was with dramatical achievement in anti-bacteria,inhibiting inflammatory,collagen deposition,angiogenesis and tissue reconstruction during the wound closure.These results indicate that the integration of traditional Chinese herbs with progressive technologies will facilitate the development and promotion of traditional Chinese medicine in modern society.

Cytological and Molecular Characteristics of Pollen Abortion in Lily with Dysplastic Tapetum

Lily was grown worldwide as a fresh cutting flower because of its colorful petals, but its anther contained a large number of pollen grains that cause serious pollen contamination, however, pollen abortion can effectively reduce the level of pollen pollution. Our analysis aims to use cytological observation to detect the critical stage when pollen abortion occurs and to provide comprehensive gene expression information at the transcriptional level. The result showed that pollen abortion in ‘Little Kiss’ began at the mononuclear stage and the callose that covers the microspores failed to degenerate when young pollens were released from the tetrads. In addition, compared with the normally developed one,the tapetum of ‘Little Kiss’ degraded in advance while the degradation of callose was delayed. Furthermore, 103 differentially expressed genes(DEGs) related to the advance degeneration of tapetum cells and callose were found in the expression levels, including 22 transcription factors(TFs). In particular, two β-glucanase genes(endo-1,3(4)-β-glucanase, exo-β-glucanase) responsible for callose degeneration were significantly down-regulated. These results suggested that pollen abortion may occur at mononuclear stage and that early degeneration of tapetum cells resulted in a significant down-regulation of β-glucanase genes. As a result, the callose to cover microspores impedes the formation of pollen walls, which may possibly lead to pollen abortion.