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Plant-associated microbes have been reported as important but overlooked drivers of plant-herbivorous insect interactions. Influence of plant-associated microbes on plant-insect interactions is diverse, including beneficial, detrimental, and neutral. Here, we determined the effects of three Penicillium fungi, including Penicillium citrinum, Penicillium sumatrense, and Penicillium digitatum, on the oviposition selection and behavior of the yellow peach moth (YPM), Conogethes punctiferalis (Guenée). Compared with fungi noninfected apples (NIA), mechanically damaged apples (MDA), and P. citrinum in potato dextrose agar medium (PC), the oviposition selection and four-arm olfactometer experiments both showed that mated YPM females preferred to P. citrinum-infected apples (PCA). For P. sumatrense or P. digitatum, we also found that mated YPM females preferred to P. sumatrense-infected apples (PSA) or P. digitatum-infected apples (PDA), respectively. Among three Penicillium fungi-infected apples, the selection rates including oviposition and olfactometer behavior of mated YPM females on PDA were both higher than those on PSA and PCA. Further analyses of host plant volatile organic compounds (VOCs) by GC-MS showed that the absolute contents of ethyl hexanoate and (Z, E)-α-farnesene in PCA, PSA, and PDA were all higher than those in NIA, and a total of 16 novel VOCs were detected in fungi-infected apples (PCA, PSA, and PDA), indicating that fungi infection changed the components and proportions of apple VOCs. Taken together, three Penicillium fungi play significant roles in mediating the host selection of YPMs via altering the emissions of VOCs. These findings will be beneficial for developing formulations for field trapping of YPMs in the future.

The present study aims to investigate the effects of ginsenoside Rg3 combined with oxaliplatin on the proliferation and apoptosis of hepatocellular carcinoma cells and the related mechanism. In this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was applied to examine the proliferation rate of hepatocellular carcinoma cell SMMC-7721 with different treatment. Flow cytometry was performed to examine apoptosis rate of hepatocellular carcinoma cells with different treatment. Immunofluorescence and Western blot methods were used to evaluate the expressions of proliferating cell nuclear antigen (PCNA) and cyclin D1 in different groups. We found that ginsenoside Rg3, oxaliplatin or ginsenoside Rg3 + oxaliplatin significantly suppressed the proliferation and promoted the apoptosis of SMMC-7721. Meanwhile, ginsenoside Rg3, oxaliplatin or ginsenoside Rg3 + oxaliplatin also significantly inhibited the expressions of PCNA and cyclin D1. Moreover, compared with ginsenoside Rg3 group and oxaliplatin group, the effect of ginsenoside Rg3 + oxaliplatin was more remarkable. Taken together, cells treated with oxaliplatin+ ginsenoside enhanced the anti-tumor effect and may inhibit the proliferation and promoted apoptosis of hepatocellular carcinoma via regulating the expression of PCNA and cyclin D1.

A novel laccase was purified from fermentation broth of white rot fungus Trametes sp. LAC-01 using an isolation procedure involving three ion-exchange chromatography steps on DEAE-cellulose, SP-Sepharose, and Q-Sepharose, and one gel-filtration step. The purified enzyme (TSL) was proved as a monomeric protein with a Mr of 59kDa based on SDS-PAGE and FPLC. Partial amino acid sequences were obtained by LC-MS/MS sharing considerably high sequence similarity with that of other laccases. It possessed optimal pH of 2.6 and temperature of 60°C using ABTS as the substrate. The Km of the laccase toward ABTS was estimated to 30.28μM at pH 2.6 and 40°C. TSL manifested considerably high oxidizing activity toward ABTS, but was avoid of degradative activity toward benzidine, caftaric acid, etc. It was effective in the decolorization of phenolic dyes - Bromothymol Blue and Malachite Green with decolorization rate higher than 60% after 24h of incubation. Adjunction of Cu(2+) with the final concentration of 2.0mmol/L significantly activated laccase production with a steady high level of 275.8-282.2U/mL in 96-144h. The high yield and short production period makes Trametes sp. LAC-01 and TSL potentially useful for industrial and environmental application and commercialization.

Peach (Prunus persica L.) is a model species for stone fruit studies within the Rosaceae family. Auxin plays an important role in the development of peach fruit. To reveal the distribution of auxin in the tissues of peach fruit, immunohistochemical localization of IAA was carried out in the seed, mesocarp, and endocarp in developing peach fruit using an anti-indole-3-acetic acid (anti-IAA) monoclonal antibody. A strong IAA signal was observed throughout the outer and inner integument during peach fruit development, and the distribution was zonal. The IAA signal was mainly focused in mucilage layers in the outer integument. The outer integument may function to produce or store IAA in the seed; a strong IAA signal was detected in the cells around the vascular tissue, whereas a weak IAA signal was located in the vascular tissues. In the mesocarp, the cells around the vascular bundle tissue gave rise to an IAA signal that increased in the late phase of fruit growth, which coincided with a significant increase in fruit growth. The distribution of IAA, however, was changed when fruit was treated with auxin transport inhibitors NPA (1-N-naphthylphthalamic acid) or TIBA (2, 3, 5-triiodobenzoic acid); in mesocarp tissues, an IAA signal was detected mainly in vessels of the t reated f ruit. During the c ritical period o f endocarp l ignification, the vessel lignification process was negatively correlated with IAA signal. The present results confirmed that the distribution of IAA was different in various tissues of peach fruit according to the developmental stage. This research provides cytological data for further study of the regulatory mechanism of auxin in peach fruit.

Raltitrexed is a specific inhibitor of thymidylate synthase (TS), which has been considered as a potential chemotherapeutic agent for the treatment of advanced gastric cancer. In the present study, the apoptosis mechanisms of raltitrexed in SGC7901 human gastric cancer cells were investigated. The cytotoxic activity of raltitrexed on SGC7901 cells was determined by cell counting kit-8 (CCK-8) assay. The CCK‑8 assay indicated that raltitrexed inhibits SGC7901 cell growth in a dose- and time-dependent manner. The morphological changes were observed by fluorescent microscopy, and characteristic morphological changes, including nuclear shrinkage and apoptotic bodies, were observed following Hoechst 33258 staining. The effects on apoptosis, cell cycle, mitochondrial transmembrane potential and reactive oxygen species (ROS) were measured by flow cytometry. The analysis revealed that raltitrexed exerted a growth inhibitory effect by inducing time-dependent apoptosis and cell-cycle arrest at the G0/G1 phase. In addition, a compromised mitochondrial membrane potential and overproduction of ROS demonstrated the involvement of the mitochondrial signaling pathway. Raltitrexed‑induced caspase‑3‑dependent apoptosis was identified using a caspase-3 activity assay and pretreatment with the caspase-3 inhibitor, Ac‑DEVD‑CHO (sequence, Ac-Asp-Glu-Val-Asp-CHO). The activity of caspase-3 was analyzed with a spectrometer. The protein expression levels of Bax, Bcl-2, cytochrome c, cleaved caspase-3 and TS were examined by western blot and the mRNA expression level of TS was detected by quantitative polymerase chain reaction. The analysis revealed that the protein levels of Bax, cytochrome c and cleaved caspase‑3 were significantly increased by raltitrexed, while Bcl-2 expression levels were reduced. Furthermore, raltitrexed increased the expression of the TS protein and mRNA in a time‑dependent manner. These results indicate that raltitrexed induces the apoptosis of SGC7901 cells through the caspase‑3‑dependent mitochondrial signaling pathway and upregulates the expression of the TS protein and mRNA.

Fruit bagging has been widely practiced in peach cultivation to produce high quality and unblemished fruit. Moreover, fruit bagging has been utilized to study the effect of shading on the quality of fruit. We conducted a proteomic analysis on peach fruit to elucidate the biochemical and physiological events that characterize the effect of bagging treatment. Comparative analysis of 2D electrophoresis (2-DE) gels showed that relative protein levels differed significantly at 125 DAFB (days after full bloom), as well as at 133 DAFB in fruit that had been bagged until 125 DAFB, followed by exposure to sunlight. Most of the proteins with altered expression were identified by MALDI TOF/TOF. Twenty-one proteins with differential expression among the groups were identified at 125 DAFB, while thirty proteins with differential expression among the groups were identified at 133 DAFB. The analysis revealed that expression of proteins involved in photosynthesis, stress responses, and biochemical processes influencing metabolism were altered during bagging treatment, suggesting that regulation of the synthesis of carbohydrates, amino acids, and proteins influenced fruit size, solid/acid ratio, and peel color. This work provides the first characterization of proteomic changes in peach in response to fruit bagging treatment. Identifying and tracking protein changes may allow us to better understand the mechanisms underlying the effects of bagging treatment.

Serological thymidine kinase 1 (STK1) is a reliable proliferation marker for prognosis, monitoring tumour therapy, and relapse. Here we investigated the use of STK1 in health screening for early detection of pre-malignant and malignant diseases. The investigation was based on 35,365 participants in four independent health screening studies in China between 2005–2011. All participants were clinically examined. The concentration of STK1 was determined by a sensitive chemiluminescent dot blot ECL assay. The ROCvalue of the STK1 assay was 0.96. At a cut-off STK1 value of 2.0 pM, the likelihood (+) value was 236.5, and the sensitivity and the specificity were 0.78 and 0.99, respectively. The relative number of city-dwelling people with elevated STK1 values (≥2.0 pM) was 0.8% (198/26,484), while the corresponding value for the group of oil-field workers was 5.8% (514/8,355). The latter group expressed significantly higher frequency of refractory anaemia, fatty liver, and obesity, compared to the city dwellers, but no cases of breast hyperplasia or prostate hyperplasia. Furthermore, people working in oil drilling/oil transportation showed higher STK1 values and higher frequency of pre-malignancies and benign diseases than people working in the oil-field administration. In the STK1 elevated group of the city-dwelling people, a statistically significantly higher number of people were found to have malignancies, pre-malignancies of all types, moderate/severe type of hyperplasia of breast or prostate, or refractory anaemia, or to be at high risk for hepatitis B, compared to people with normal STK1 values (< 2.0 pM). No malignancies were found in the normal STK1 group. In the elevated STK1 group 85.4% showed diseases linked to a higher risk for pre-/early cancerous progression, compared to 52.4% of those with normal STK1 values. Among participants with elevated STK1 values, 8.8% developed new malignancies or progress in their pre-malignancies within 5 to 72 months, compared to 0.2% among people with normal STK1 values. People who showed elevated STK1 values were at about three to five times higher risk to develop malignancies compared to a calculated risk based on a cancer incidence rate of 0.2–0.3%. We conclude that serological TK1 protein concentration is a reliable marker for risk assessment of pre/early cancerous progression.

Helicoverpa armigera is one of the most harmful pests in China. Although it had been successfully controlled by Cry1A toxins, some H. armigera populations are building up resistance to Cry1A toxins in the laboratory. Vip3A, secreted by Bacillus thuringiensis, is another potential toxin against H. armigera. Previous reports showed that activated Vip3A performs its function by inserting into the midgut brush border membrane vesicles (BBMV) of susceptible insects. To further investigate the binding of Vip3A to BBMV of H. armigera, the full-length Vip3Aa10 toxin expressed in Escherichia coli was digested by trypsin or midgut juice extract, respectively. Among the fragments of digested Vip3Aa10, only a 62kDa fragment (Vip3Aa10-T) exhibited binding to BBMV of H. armigera and has insecticidal activity. Moreover, this interaction was specific and was not affected by the presence of Cry1Ab toxin. Binding of Vip3Aa10-T to BBMV resulted in the formation of an ion channel. Unlike Cry1A toxins, Vip3Aa10-T was just slightly associated with lipid rafts of BBMV. These data suggest that although activated Vip3Aa10 specifically interacts with BBMV of H. armigera and forms an ion channel, the mode of action of it may be different from that of Cry1A toxins.

The present experiment investigates the effect of neostigmine on nicotinic acetylcholine receptors (nAChRs) in the cultured neurons from neonatal rat superior cervical ganglia (SCG). Using whole-cell patch clamp techniques, we found that the amplitudes of the currents induced by 50 microM dimethylphenylpiperazinium (DMPP) were 21.5+/-10.7%, 52.9+/-9.2% and 86.9+/-4.9% depressed at the increased concentrations of neostigmine 100, 200 and 400 microM, respectively. The inhibition of neostigmine decreased gradually with the increased concentration of nicotine from 10 to 160 microM. Lineweaver-Burk's double-reversible plot illustrated that neostigmine blocked neuronal nAChRs in a competitive manner. Hyperpolarization of membrane potential from -40 mV to -100 mV did not significantly influence the blockade of neostigmine. Neostigmine could not accelerate the decay of the DMPP-induced currents, neither evoke any detectable currents in SCG neurons. The results indicate that neostigmine depress neuronal nAChRs in a competitive, concentration-dependent and voltage-independent manner, and can not facilitate desensitization of the receptors. The present data suggest that neostigmine blocks neuronal nAChRs by interacting with the ACh binding sites of the receptors.

NAC transcription factor is a new-found and multifunctional transcription factor in plants. The purpose of the paper is to study the relative expression trends of BnNAC transcription factor gene under cold (4°C), salinity (200 mM NaCl) and abscisic acid (ABA) (100 μM) stress in Brassica napus. The functional region of BnNAC gene was cloned through the homologous cloning method. The sequences of nucleic acid and amino acid were analyzed using BLAST and DNAMAN software. Relative expression trends of BnNAC gene were detected by applying real-time relative quantification PCR (RT-qPCR) under cold, salinity and ABA treatment. A 440 bp BnNAC gene segment was acquired in Brassica napus. BnNAC gene expressions of Brassica napus seedlings were induced under cold, salinity and ABA treatment conditions. The results of RT-qPCR analysis revealed that, under salinity, cold and ABA conditions, the relative expression of BnNAC gene increases with time, reaches the peak at 24h, 3h, 12h respectively and then decreases. Our research showed that BnNAC plays a positive role in salinity, cold and ABA stress response.

The development of the stone and formation of peach (Prunus persica) fruit were explored in this work using a proteomic approach. Sixty-eight proteins with different expression patterns were identified in both the endocarp and mesocarp during early fruit development (from 28 to 59 days after flowering) and the majority were involved in primary or secondary metabolism. In contrast to most proteins associated with primary metabolism in the endocarp, whose expression is down-regulated, expression of pyruvate dehydrogenase (PDH) unexpectedly increased exponentially. Moreover, its expression pattern was linearly positively correlated with the exponentially growing lignin content (R = 0.940), which suggests that PDH may play a role in endocarp lignification. Our data also revealed different spatiotemporal expressions of enzymes involved in the lignin and flavonoid pathways that provided proteome-level evidence to support the hypothesis that these two pathways are competitive during endocarp development. In addition, we observed endocarp-specific oxidative stress and propose that it may act as a stimulating factor in activating lignification and subsequent programmed cell death in the endocarp.

To investigate the expressions of chemokine receptors and interleukin (IL) receptors on the peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients and their correlations with clinical features as well as SLE disease activity index (SLEDAI).The mRNA expressions of chemokine receptors and IL receptors on PBMCs of 93 SLE patients and 30 healthy controls were detected by reverse transcription-polymerase chain reaction, including CCR2, CCR3, CCR4, CCR5, CCR6, CCR8, CXCR3, CXCRS, CX3CR1, XCR1, IL-4R, and IL-10R. The clinical features of SLE patients were recorded. The correlations of chemokine receptors and IL receptors mRNA expressions with clinical features as well as SLEDAI were assayed using linear regression analysis.The level of CCR5 mRNA in SLE patients (including active and inactive SLE) was significantly higher than that in healthy controls (P0.05), and there was no significant difference between active and inactive patients in this respect (P0.05). CX3CR1 mRNA expression significantly increased from healthy control to inactive SLE to active SLE in sequence. The others (except for CCR8, CXCR3, and IL-10R) in active SLE patients were significantly higher than those in both inactive SLE patients and healthy controls (all P0.05). There were positive correlations between SLEDAI and CCR2 (r = 0.424, t = 4.313, P0.001), CCR3 (r = 0.518, t = 5.410, P0.001), CCR4 (r = 0.376, t = 3.851, P0.001), CCR6 (r = 0.457, t = 4.513, P0.001), CXCR5 (r = 0.455, t = 4.629, P0.001), CX3CR1 (r = 0.445, t = 4.523, P0.001), as well as XCR1 (r = 0.540, t = 5.445, P0.001). And CCR5 mRNA expression level was positively correlated with IL-4R mRNA (r = 0.313, t = 2.353, P0.05). The patients with myositis and cutaneous vasculitis simultaneously showed lower levels of CCR5 and CX3CR1, and CCR5 expression was negatively correlated with the scores of SLEDAI in SLE cases accompanied by photosensitivity (r = 0.426, t = -2.155, P0.05).Increased expressions of CCR5 and CX3CR1 on PBMCs may be indicators in clinical survey for SLE.

Eighty eight accessions including eighty six of Solanum melongena and two wild related species (Solanum integrifolium and Solanum torvum) were used to analyze the genetic variation with 23 SSR primers. And 20 primers were successfully amplified, of which 19 showed polymorphism with a total of 68 detected alleles. The number of alleles per primer ranged from 2 to 6, with a mean of 3.6. Among 19 polymorphism primers, four primer pairs (EM117, EM155, EM126, EM127) gave rise to multiple polymorphic bands, two of which could be used for varieties certification of eggplant. The average genetic diversity of all accessions measured by the Nie's genetic diversity, Shannon's Information index and Effective number of alleles were 0.41(range 0.12~0.80), 0.7443(rang from 0.2617~1.6958) and 2.0151(ranged from 1.1307~5.310), respectively. Genetic similarity among all the accessions ranged from 0.09 to 1 with an average of 0.69. The high diversity was displayed. Additionally, S. melongena had high average similarity with the wild species S. integrifolium (0.61) and low average similarity with S. torvum (0.26). UPGMA clustering at genetic similarity value of 0.60 grouped all the eggplant accessions into four groups, where S.torvum formed a separate and more distant cluster. Moreover, the fourth group can be further classified into two sub-groups, and the wild species S. integrifolium can be clearly separated from the other cultivated eggplants. However, majority of the cultivated eggplants were not clustered in ways concordant with taxonomic information such as collection sites, fruit shape or any other known trait. The genetic diversity and phylogenetic relationships of eggplant accessions reported in this study might be used for breeder when planning future crosses between cultivated eggplant and the wild species.

Deterioration of peach fruit quality because of split-pit formation had bad influence on market (1). Though many studies have mentioned the mechanism of split-pit formation, further studied are still needed to uncover the development mechanism of endocarp cell during pit hardening process, especially the Programmed cell death (PCD) process of peach endocarp cell. Here, the development of the peach endocarp cell from the two Chinese peach varieties "Jingyu" and "Okubo" was detailedly studied to understand the PCD of endocarp cell during the lignin formation process. According to our morphology observation using transmission electron microscopy, the PCD process of the endocarp cell, characterized by vacuolar enlargement, might have been initiated as early as eight days after full athesis (maybe even more earlier), which is earlier than that of the split-pit formation process. In our studies, almost all the endocarp cell organelles were found to degenerate at forty days after full athesis, when thickening of the endocarp cell wall was also observed. Furthermore, the involvement of hydrolytic enzymes, in particular proteases and nucleases, in the autolytic process of PCD of peach endocarp cells was quantitatively analyzed during pit hardening process. The peach endocarp protein was found to be rapidly degraded by newly expressed proteases since 35 days after full athesis. Elevation of nuclease activities was found to be also coupled with PCD of peach endocarp cells. Furthermore, DNA degradation was found to happen earlier than RNA degradation and hydrolyzing of endocarp soluble protein during the peach pit hardening process.

Few studies were mentioned to illustrate the role of plant hormones-IAA and ethylene in split-pit formation. Here, a random sample of 100 developing fruits from 7-year-old early maturing nectarines (Prunus persica L. Batsch cv. Ruiguang 5) was used as test material. The shape index and growth index of the sampled nectarines was comparatively analyzed to show the relation between fruit un-uniform growth and their split-pit formation. The split-pit fruits were found to have a higher degree of un-uniform growth than that of the normal fruits, which were demonstrated by higher ratio of transverse diameter/lateral diameter and bigger difference between the flesh thickness of the two sides in the middle part of the fruits (along the line of the transverse diameter) about twelve weeks after full anthesis (p < 0.05). Subsequently, a higher degree of unbalanced distribution of IAA and especially ethylene was observed in the split-pit fruits compared to the normal fruits, which may correlate with un-balanced expansion of the nectarines flesh. Higher concentration of IAA and especially ethylene was found in the middle part of the split-pit fruits than those in the sutures, while those in the normal fruits were not significant. Furthermore, the caliosity was specifically observed in the internal surface of the split-pit nectarines endo carp as early as four weeks after full anthesis, but not in the normal nectarines. The occurrence of caliosit in the endo carp may be an early signal of spilt-pit.

With four-year-old potted Prunus persica L. cv. Qingfeng as test material, this paper studied the change pattern of its leaf betaine content under water stress, and its physiological responses under effects of foliage-spraying exogenous betaine. The results showed that under normal water supply, the betaine content in Qingfeng' s leaf was 75.9-80.5 microg x g(-1) FM, which was increased with increasing water stress, and up to 278.9 microg x g(-1) FM on the 16th day after cutting off the water supply. The leaf plasma membrane permeability was 8.06% - 8.61% under normal water supply, but increased to 28.62% under water stress. When 100 and 500 mg x L(-1) of betaine were applied exogenously, the plasma membrane permeability was 26.25% and 21.79% after 16 days, respectively. The hydrogen peroxide (H2O2) content increased from 27.2-32.5 micromol g(-1) FM to 76.4 micromol x g(-1) FM in the course of water stress, and decreased to 73.2 and 68.5 micromol x g(-1) FM after spraying 100 and 500 mg betaine x L(-1), respectively. During the period of intensified water stress, the peak value of ascorbate peroxidase (AsA-POD) activity was 0.435 mg x g(-1) FM, and up to 0.490 mg x g(-1) FM when treated with exogenous betaine. When the peach tree was subjected to water stress, the contents of free proline and soluble sugar accumulated dramatically, but produced on approximately decrease in 500 mg x L(-1) endogenous betaine application on the 16th day which was slightly less than that of control and 100 mg x L(-1) betaine application. There was a gradual decline in the content of soluble protein under water stress, and an increment of 20. 3% was observed when betaine was applied exogenously. These results strongly suggested that foliage-spraying exogenous betaine could increase the drought resistance of peach tree through decreasing its leaf plasma membrane permeability and H2O2, free proline and soluble sugar contents and increasing its leaf AsA-POD activity and soluble protein content.

To detect the methylation status of p16 gene promotor in DNA derived from plasma and blood cells of patients with systemic lupus erythematosus (SLE) , and it's relationship with clinical symptoms.p16 promotor methylation in plasma and peripheral blood cells (PBCs) DNA were simultaneously detected with the methylation specific PCR (MSP) method in 24 active SLE patients, 21 inactive SLE patients, as well as 20 healthy controls.In the plasma DNA, p16 gene methylation ratio (MP%) was higher in SLE patients than in the healthy controls (64.4% vs. 5.0%, P0.05). MP% in the active SLE patients was significantly higher than that in the inactive SLE patients (83.3% vs. 42.9%, P0.05). In the PBCs, p16 gene methylation ratio (MC%) in the healthy controls was significantly higher than that in SLE (80.0% vs. 48.9%, P0.05). MC% in the active SLE patients (29.2%) was the lowest among three groups. There was no significant difference between the inactive SLE patients and healthy controls (71.4% vs. 80.0%, P0.05). Each patient could be judged as one of the four methylation patterns: MP/MC, UP/MC (UP: unmethylated plasma p16) , MP/UC (UC: unmethylated PBCs p16) , and UP/UC. The ratios of MP/ MC and UP/UC were similar between the active and inactive SLE patients. However, different distributions of other two patterns were found in the active and inactive SLE patients as UP/MC 4.2% vs. 42.9% (P0.05) and MP/UC 58.3% vs. 14.3% (P0.05), respectively. The active SLE patients with MP/UC and the inactive SLE patients with UP/MC showed different clinical symptoms and laboratory examinations. Significant correlation was found between the disease activity index for lupus patients (SLEDAI) scores and MP% (r = 0.93), between the SLEDAI scores and MC% (r = - 0.96) also between MC% and MP% (r = - 0.79).The p16 methylation assay provides available information for the diagnosis, judgment of disease activity, as well as novel insights into the pathogenesis underlying this disease.