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Anxiety-like behaviors in mice include social avoidance and avoidance of bright spaces. Whether these features are distinctly regulated is unclear. We demonstrate that in mice, social and anxiogenic stimuli, respectively, increase and decrease serotonin (5-HT) levels in basal amygdala (BA). In dorsal raphe nucleus (DRN), 5-HT∩vGluT3 neurons projecting to BA parvalbumin (DRN

Major depressive disorder is a devastating psychiatric disease that afflicts up to 17% of the world's population. Postmortem brain analyses and imaging studies of patients with depression have implicated basal lateral amygdala (BLA) dysfunction in the pathophysiology of depression. However, the circuit and molecular mechanisms through which BLA neurons modulate depressive behavior are largely uncharacterized. Here, in mice, we identified that BLA cholecystokinin (CCK) glutamatergic neurons mediated negative reinforcement via D2 medium spiny neurons (MSNs) in the nucleus accumbens (NAc) and that chronic social defeat selectively potentiated excitatory transmission of the CCKBLA-D2NAc circuit in susceptible mice via reduction of presynaptic cannabinoid type-1 receptor (CB1R). Knockdown of CB1R in the CCKBLA-D2NAc circuit elevated synaptic activity and promoted stress susceptibility. Notably, selective inhibition of the CCKBLA-D2NAc circuit or administration of synthetic cannabinoids in the NAc was sufficient to produce antidepressant-like effects. Overall, our studies reveal the circuit and molecular mechanisms of depression.

Differences in the gene mutation spectra of younger and older Chinese adult AML patients and the prognostic significance of these differentially presented gene mutations are rarely reported. One hundred and thirteen newly diagnosed Chinese adults with AML, divided into groups of younger and older patients, were enrolled in this study. Bone marrow samples from the patients were analyzed using targeted next-generation sequencing with a panel of 141 genes. Ninety-eight mutated genes were detected and the top 10 mutated genes were KMT2D, FLT3, FAT1, ASXL1, NRAS, DNMT3A, RELN, TET2, JAK2, and KRAS. The top five functional groups were the tyrosine kinase pathway, transcription factors, DNA methylation, chromatin modifiers, and the JAK-STAT signaling pathway. Younger patients exhibited higher incidences of KMT2D (33.8% vs 10.4%, P = 0.004) and KRAS (15.4% vs 2.1%, P = 0.042) mutations than older patients; whereas, older patients harbored more SRSF2 (20.8% vs 0%, P = 0.002), transcription factor (85.4% vs 67.7%, P = 0.031), DNA methylation (58.3% vs 36.9%, P = 0.024), and RNA splicing (31.3% vs 12.3%, P = 0.013) mutations than younger patients. Moreover, patients with SRSF2 mutations exhibited a lower rate of overall survival (P 0.001) and relapse-free survival (P 0.001) than patients carrying wild-type SRSF2. In conclusion, rarely reported KMT2D, FAT1, and RELN mutations were detected at high frequencies in our cohort. The gene mutation spectrum of older patients was different to that of younger patients. Moreover, older patients harbored more SRSF2 mutations, which predicted lower rates of overall and relapse-free survival.

Major depressive disorder is a devastating psychiatric disease that afflicts up to 17% of the world's population. Postmortem brain analyses and imaging studies of patients with depression have implicated basal lateral amygdala (BLA) dysfunction in the pathophysiology of depression. However, the circuit and molecular mechanisms through which BLA neurons modulate depressive behavior are largely uncharacterized. Here, in mice, we identified that BLA cholecystokinin (CCK) glutamatergic neurons mediated negative reinforcement via D2 medium spiny neurons (MSNs) in the nucleus accumbens (NAc) and that chronic social defeat selectively potentiated excitatory transmission of the CCK

In order to investigate the anti-aging effect and mechanism of Liuwei Dihuang Wan pill (LWDH) on natural aging rats, LWDH pill was abstracted by boiling and filtrating according to conventional method and became decoction. Adult male rats were divided into two groups randomly, the experimental group fed with decoction of LWDH (0.5 mg/ml/100 g body weight), and natural aging group fed with equivalent volume of tap water. Both groups were treated with respective material once each day for one year. Morris water maze was employed to examine the behavioral learning and memory ability of all rats in the experiment. The content of superoxide dismutase (SOD), an antioxidant, and acetylcholinesterase (AChE) in serum were tested with spectrophotometer. The activity of choline acetyltransferase (ChAT) in brain cortex was assayed by spectrophotometer. The thymus index and spleen index were analyzed. The results showed that LWDH can improve general behavior of aging rats. Compared with the control group, the weight of natural aging rats increased stably and was much higher (P ＜ 0.05). The LWDH-treated rats showed a shorter average escape latency in the navigation test (P ＜ 0.05) and higher percentage of swimming time spent in the target quadrant of Morris water maze and later spatial probe test (P ＜ 0.05). Higher thymus index and spleen index (P ＜ 0.05), increased SOD vigor in serum (P ＜ 0.05) as well as higher ChAT and AChE in cerebral cortex (P ＜ 0.05) were also observed. As above, LWDH can improve learning and memory ability and general behavior of natural aging rats; it can also reverse the metabolic capability and immunoregulation in natural aging rats.

Finite element analysis and fuzzy multi-objective optimization technique have been integrated to solve the die structure design of sheet metal forming by transforming fuzzy multi-objective problem into a normal optimization problem. A mathematical model of fuzzy optimization for bending die was established. A fuzzy goal set was constructed. The fuzziness of multi-objective functions and constraints were defined. The optimal solution and optimal constraint value of individual objective function in the feasible field were found using the genetic algorithms. An electronic part bending case shows that this approach is more effective and accurate than traditional finite element analysis method and the ‘trial and error’ procedure.

In the past 50 years, many long-span continuous stone arch bridges have been built in China. Analysis of mechanical performances and load capacities of long-span continuous stone arch bridges has important significances for the safety assessment of the similar bridges. 3D elastoplastic finite element method is employed to analyze ultimate bearing capacity of a three-span arch bridge with sandstone masonry in this paper. The characteristics of structural geometric and material nonlinearities and cracking and crushing of the masonry are taken into account. Compared with single-span arch bridge, both the coaction of spandrel structure and main arch and the influence of loading arrangements on ultimate bearing capacity is analyzed. The failure characteristics of the structure under the ultimate load are also introduced. The analysis results and conclusions can be referenced for the safety assessment of similar bridges.

In recent years, the internal combustion engine has been widely used through technological advances to improve its environmental performance. Mechanical and electrical integration of the engine turbocharging system is based on conventional turbocharging system to increase motor in parallel with the turbocharger and the corresponding reversible energy storage components, so that achieve by adjusting the energy input or output direction and the size of the motor to adjust the exhaust turbocharger operating point and the gas supply function. According to matching requirements of light vehicle diesel engine, the analysis model of exhaust gas energy is obtained through qualitative analysis of exhaust gas energy in turbocharged diesel engine.

In the version of this article originally published, there were several errors in Fig. 4. In Fig. 4a, the title read ‘3D repeated optical inhibition after CSDS.’ It should have read ‘3-day repeated optical inhibition after CSDS.’ In Fig. 4c, two labels that should have been aligned with the time axis appeared in the wrong place in the figure. The ticks labeled ‘SI’ and ‘Fiber implant’ should have also been labeled with ‘10’ and ‘14,’ respectively. Additionally, in Fig. 4j, a label that should have been aligned with the time axis appeared in the wrong place in the figure. The tick labeled ‘Fiber implant’ should have also been labeled with ‘14.’ The errors have been corrected in the print, PDF and HTML versions of the manuscript.

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important wheat diseases in many re- gions in the world. Breeding for resistance is the most economical and effective method for controlling this disease. Wild emmer (Triticum turgidum var. dicoccoides) is the immediate progenitor of cultivated tetraploid and hexaploid wheats and has been proven to be an important resource of disease resistance improvement. In this study, a common wheat line 3D258 with powdery mildew resistance was developed by crossing wild emmer accession WE29 with common wheat landrace Yanda1817 and back- crossing with common wheat line 87-1 (Yanda 1817/WE29//5*87-1, BC4F6). Genetic analysis of the F2 population and their F3 families, developed from 3D258 and a susceptible common wheat cultivar Xuezao, indicated that the powdery mildew resistance in 3D258 was controlled by a single dominant gene, designated temporarily as MlWE29. Molecular markers and the bulked seg- regant analysis were used to characterize the powdery mildew resistance gene MlWE29. Six SSR markers (Xgwm335, Xgwm213, Xgwm639, Xwmc415, Xwmc289, and Xwmc75) and five EST-STS markers (BE494426, BE442763, CD452476, BE445282, and BE407068) were found to be linked to MlWE29. Using Chinese Spring nullisomic-tetrasomics, ditelosomics and deletion lines

This study was aimed to investigate the protective effect of Liu Wei Dihuang (LWDH) against D-galactose (D-gal)-induced brain injury in rats and the existence of sex-dependent differences in LWDH protection. Sixty-four rats evenly composed of males and females were randomly assigned into 4 groups (n = 8): normal saline (NS) + NS (N + N), NS + LWDH (N + L), D-gal + NS (D + N) and D-gal + LWDH (D + L) groups. Rats in D + N and D + L groups received daily injection of D-gal (100 mg/kg, s.c.) for six weeks to establish the aging model, while rats in N + N and N + L groups were injected with the same volume of NS. From the third week, rats in N + L and D + L groups were orally administered with a decoction of LWDH for subsequent six weeks. Rats in N + N and D + N groups were orally administered just with the same volume of NS simultaneously. Morris water maze test was employed to evaluate the ability of learning and memory of the rats in all the groups. Acetylcholine (ACh) content, activities of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) in visual cortex were assayed. Hematoxylin and eosin (HE) staining were used to observe the morphologic injury in hippocampus and visual cortex, and immunohistochemistry was performed to evaluate ChAT and AChE expression levels in the visual cortex. The results showed that the rats in D + N groups exhibited a longer escape latency to platform, lower swimming speed, less percent of target quadrant search time and platform crossings, compared with N + N groups, suggesting the establishment of aging model, while LWDH improved these indexes in D-gal-treated rats. Compared with D + N groups, LWDH increased ACh content and ChAT activity, and decreased AChE activity in visual cortex. Remarkable loss of neurons was found in hippocampus and visual cortex of aging rats, and the injury was significantly attenuated by LWDH. Immunohistochemistry showed D-gal-induced decreases of ChAT and AChE expressions were restored by LWDH. Furthermore, under the neural protection of LWDH, the improvement on platform crossings in male aging rats was better than that in female ones, while in ChAT expression and neuron density in visual cortex, female aging rats obtained more amelioration. These results suggest LWDH can markedly reverse the D-gal-induced cognitive impairments and neuronal damage in both hippocampus and visual cortex, which are achieved at least partly through restoring cholinergic system in central nervous system. Moreover, there is some sex difference in protective effects of LWDH against D-gal-induced impairment.

To investigate the effect of bortezomib alone or combined with harringtonine (HT) or arsenic trioxide (As2O3) on the proliferation capacity and apoptosis of HL-60/ADM cell line and fresh cells from refractory/relapse acute leukemia patients.HL-60/ADM cells or refractory/relapse leukemia cells were incubated with bortezomib at different doses alone and in combination with HT or As2O3. The proliferation capacity was observed by MTT assay, cell apoptosis by fluorescence microscopy and flow cytometry. Intracellular concentration of daunorubicin (DNR) was determined by flow cytometry.In bortezomib-treated HL-60/ADM cells, the proliferation inhibition rate and apoptotic cells increased in a time- and dose-dependent manner. 40 nmol/L bortezomib could maximally inhibit the proliferation of HL-60/ADM cells at 48 hours. 15 micromol/L As2O3 or 752 nmol/L HT combined with different doses of bortezomib could inhibit proliferation and induce apoptosis of HL-60/ADM cells. The As2O3 plus bortezomib or HT plus bortezomib showed a greater anticancer efficacy than either of the drugs alone (P0.05, P0.01). Bortezomib (10 nmol/L) could markedly enhance the intracellular accumulation of DNR in HL-60/ADM cells (P0.05).Bortezomib can inhibit proliferation and induce apoptosis of HL-60/ADM cells and fresh refractory/relapse acute leukemia cells, especially combined with HT or As2O3.

Wheat cultivar Jing 411 which is susceptible to powdery mildew, wheat cultivar Brock and near isogenic lines (NILs) of Jing411, which are resistant to powdery mildew were analysized for polymorphisms using 225 pairs of AFLP primers Only two pairs of primers Pst+GAC/Mse+ TCT (P1) and Pst+AGC/Mse+ACC (P2) stably produced polymorphic bands between the resistant and susceptible plants. Two specific fragments were obtained. By cloning and sequencing these two specific fragments, it was showed that the specific fragment amplified by primer P1 had 268bp, and the fragment amplified by P2 had 227bp. They were named AFLP marker P1(268) and P2(227) respectively. Linkage analysis of these two markers revealed that the polymorphism existed in a 106 F2 segregating population. These two markers closely linked to a powdery mildew resistance gene in wheat cultivar Brock, linkage distance were 3.6 and 1.9 cM respectively. These two markers will be useful for marker-assisted selection and gene pyramiding in wheat resistance breeding.

The availability of high-throughput genotyping technologies and microarray assays has allowed researchers to investigate genetic variations that influence levels of gene expression. Expression Quantitative Trait Locus (eQTL) mapping methods have been used to identify the genetic basis of gene expression. Similar to traditional QTL studies, the main goal of eQTL is to identify the genomic locations to which the expression traits are linked. Although microarrays provide the expression data of thousands of transcripts, standard QTL mapping methods, which are able to handle at most tens of traits, cannot be applied directly. As a result, it is necessary to consider the statistical principles involved in the design and analysis of these experiments. In this paper, we reviewed individual selection, experimental design of microarray, normalization of gene expression data, mapping methods, and explaining of results and proposed potential methodological problems for such analyses. Finally, we discussed the applications of this integrative genomic approach to estimate heritability of transcripts, identify candidate genes, construct gene networks, and understand interactions between genes, genes and environments.

To study the effect of granulocyte colony-stimulating factor (G-CSF) on the proliferation and differentiation of bcr/abl(+)-CD34+ cells.bcr/abl(+)-CD34+ cells were isolated from bone marrow of chronic myelocytic leukemia (CML) patients and were treated with 0, 10, 100, 1000 ng/ml of G-CSF for 48, 96, 144 hs. CD34 cells from normal bone marrow were used as controls. Cell proliferation was determined by trypan blue dye exclusion, cell-cycle and antigen differentiation were determined by flow cytometry and cell morphology was observed under light microscope.The number of bcr/abl(+)-CD34+ cells was increased obviously in all groups. After cultured for 48 and 96 h, the number of bcr/abl(+)-CD34+ cells at G-CSF 10 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P0.05) , the number of normal CD34 cells was increased only in the presence of G-CSF. After cultured for 48, 96 and 144 h, the cell number in G-CSF 100 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P0.05, P0.01, P0.01, respectively). After cultured for 144 h, the cell percentages in G0/G1 phase for bcr/abl(+)-CD34+ cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that in G-CSF 0 ng/ml group (P0. 05), and that for normal CD34 cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that of G-CSF 0 ng/ml group after cultured for 48 and 96 h. The expressions of CD34 on bcr/abl(+)-CD34+ cells and normal CD34+ cells were decreased along with the culture duration, accompanied by the expression of CD33 and CD13 increased first and decreased later, which was not correlated with the concentration of G-CSF. Both bcr/abl(+)-CD34+ cells and normal CD34+ cells showed mature morphology along with proliferation and differentiation.G-CSF promotes proliferation of both bcr/abl(+)-CD34+ cells and normal CD34+ cells, but not necessary for the former, and the former differentiates more rapidly than the latter does, but both was independent of G-CSF.

To study the effects of STI571, arsenic trioxide (As2O3) and Velcade, used alone or in combination, on the proliferation and apoptosis of bcr/abl+-CD34+ cells.bcr/abl+-CD34+ cells isolated from the bone marrow of patients with chronic myeloid leukemia (CML) were treated for 96 h with STI571, As2O3 and Velcade either alone and in combination, and the cell proliferation and apoptosis were analyzed by CCK-8 assay and flow cytometry. The morphological changes of the apoptotic cells were observed by Hoechst33342 staining and fluorescent microscope. The inhibitory effects of the drugs on normal CD34+ cells were also observed.Low-concentration STI571, As2O3 or Velcade all dose-dependently inhibited bcr/abl+-CD34+ cell proliferation without obvious apoptosis-inducing effects. STI571 at 0.25-2 micromol/L combined with As2O3 at 2.5 micromol/L and with Velcade at 15 nmol/L both significantly increased the cell inhibition and apoptosis rates, showing obvious additive or synergistic effects of the drugs without further enhancement of normal CD34+ cell inhibition.Combination with STI571 enhances the effects of As2O3 and Velcade on bcr/abl+-CD34+ cells, suggesting the potential clinical value of this regimen.

To assess the antitumor efficacy and adverse effects of bortezomib either used alone or in combination with arsenic trioxide for transplanted tumor in nude mice.Nude mice bearing HL-60 cell xenografts were randomized into 4 groups to receive treatment with normal saline, bortezomib, arsenic trioxide, bortezomib plus arsenic trioxide. The tumor growth inhibition and general condition of the nude mice were observed, and in situ TUNEL assay and immunohistochemistry were performed on the transplanted tumors.Bortezomib alone and in combination with arsenic trioxide could both inhibit the growth of the transplanted tumors, prolong the survival of the nude mice, and induce cell apoptosis and growth inhibition of the HL-60 cells in vivo, and the combined administration exhibited even better effects. The administration was well tolerated with causing manifest vital organ damages in the mice.Bortezomib in combination with arsenic trioxide has significant antitumor effect in nude mice bearing HL-60 cell xenografts possibly by inducing HL-60 cell apoptosis and growth inhibition without producing no significant adverse effects.

To investigate the effect of bortezomib alone and in combination with arsenic trioxide on apoptosis of HL-60 cells.HL-60 cells were treated with bortezomib alone or in combination with arsenic trioxide for 12 to 48 h and the cell proliferation was analyzed with MTT assay, and cell apoptosis detected by DNA gel electrophoresis, fluorescence microscopy and flow cytometry.At the concentrations of 10 to 50 nmol/L, bortezomib effectively inhibited HL-60 cell proliferation, and induced cell apoptosis. A 12-hour bortezomib treatment at 10 nmol/L was sufficient to induce cell apoptosis, and prolonged treatment or increased concentration significantly increased HL-60 cell apoptotic rate. Combined treatment of the cells with bortezomib (10 nmol/L) and arsenic trioxide (15 micromol/L) resulted in an even higher cell apoptosis rate than that induced by the respective agent alone.Bortezomib can induce HL-60 cell apoptosis in a time- and dose-dependent manner, and a synergistic effect can be observed of bortezomib and arsenic trioxide in apoptosis induction.

We reported previously that C-fibers innervating rat skin can be excited by short trains of electrical shocks ('tetanus') applied to neighboring nerves. Since these nerves were disconnected from the CNS, the cross-talk is located peripherally. Here we tested if low-threshold mechanoceptive (LTM) C-fibers can be excited by this cross-talk and if this process is mediated by substance P (SP) via neurokinin-1 (NK-1) receptors. In urethane anesthetized rats we found that 80% (56/71) of LTM C-fibers, recorded in the lateral cutaneous branch of the dorsal ramus (CBDR) of T10 spinal nerve, were excited by a 10s, 20 Hz tetanus of the T9 CBDR. Compared to the spontaneous pre-tetanic firing frequency of 1.62+/-0.40 impulses/30s, the frequency significantly increased to 3.74+/-0.99, 3.17+/-0.69 and 2.92+/-0.63 impulses/30s, at 30, 60 and 90 s after the tetanus, respectively, and declined to the baseline frequency thereafter. When injected into their receptive fields, SP mimicked the tetanically induced increase of firing rate, whereas the NK-1 receptor antagonist WIN 51708 blocked the excitation in most fibers. The excitation was significantly diminished in adult rats that were neonatally treated with capsaicin, a treatment that destroys most SP-expressing afferent fibers. Thus, we conclude that peptidergic primary afferents are functionally linked with adjacent LTM C-fibers in a non-synaptic, paracrine-like signaling pathway via SP and NK-1 receptors, and perhaps also other agents as well. We propose that this cross-talk has evolved as a mechanism regulating the mechanoceptive characteristics of LTM C-fibers, presumably contributing to pain sensation elicited by tactile stimuli ('allodynia').

The aim of this study was to investigate the effect of bortezomib alone and in combination with harringtonine on apoptosis of HL-60 cells. HL-60 cells were treated with bortezomib, harringtonine in different concentrations for 12 - 48 hours. Cell proliferation was analyzed by MTT assay; the apoptosis of HL-60 cells was observed by DNA gel electrophoresis, fluorescence microscopy and flow cytometry. The results showed that 10 - 50 nmol/L bortezomib could effectively inhibit HL-60 cell proliferation, and induced its apoptosis. After treating for 12 hours, 10 nmol/L bortezomib could trigger cells apoptosis. With time prolongation or dose increase, HL-60 cell apoptotic rate significantly increased. Furthermore, co-administration of bortezomib (10 nmol/L) with harringtonine (30 nmol/L) resulted in a higher cell apoptotic rate when compared with that induced by those agents used alone. It is concluded that the bortezomib can induce HL-60 cells apoptosis in a time-and-dose-dependent manner and synergistic effectiveness can be found when bortezomib combined with harringtonine.

Comparative studies of chloroplast proteome on different developmental stage (seeding, tillering, shooting, booting stage) of leaves have been made in isoplamic allonuclear male-sterile lines Nongda 3237A, Xiaoyan No.6 A and their maintainer lines by 2D-PAGE. The results indicated that no obvious differences were found in chloroplast proteome between Nongda 3237A, Nongda 3237B, Xiaoyan No.6A, Xiaoyan No.6B at different developmental stages of leaves. Differences were found just at booting stage in Xiaoyan No.6A and its maintainer lines. Obvious differences, however, were observed in chloroplast proteome between isoplasmic allonuclear male-sterile lines Nongda 3237A and Xiaoyan No.6A. For instance,2 protein spots (pl5.4/34kDa, pl5.4/32kDa) at seedling stage,6 protein spots (pl5.4/80kDa, pl5.4/65kDa, pl5.4/60kDa, pl5.4/48kDa, pI5.4/40kDa, pI5.4/35kDa) at booting stage were present in Xiaoyan No.6A and absent in Nongda 3237A. pl6.3/18kDa was present at seedling stage in Nongda 3237A and absent in Xiaoyan No. 6A. pI6.8/28kDa protein spot revealed the developmental changes. It was present in leaves at seedling, tillering, shooting stages and absent at flowering stage in Nongda 3237A. No development changes of the protein spot were observed in Xiaoyan No.6A. These experiment results demonstrated that it was possible chloroplast proteome wasn't relative to the cytoplasmic male-sterile characteristics. But nuclear background in male -sterile lines can obviously affect chloroplast protein composition. Distant relative on nuclear-cytoplasmic has larger differences on chloroplast proteome than close relative on nuclear-cytoplasmic in CMS.

To explore the mechanism of apoptosis of myeloid leukemia cells induced by Bortezomib, a proteasome inhibitor.Human acute myeloid leukemia cells of the line HL60 were cultured and treated with Bortezomib of the concentrations of 0, 10, 20, 30, and 40 micromol/L for 24 hours. MTT assay and flow cytometry were used to detect the proliferation inhibition and apoptosis. Hoechst 33342 staining was used to observe the morphology of the cells. Western blotting was used to detect the protein expression of Bcl-2, Caspase-9, Caspase-3, and poly ADP-ribose polymerase (PARP).Bortezomib could induce HL60 cell apoptosis dose- and time-dependently. After treated for 24 hours by 30 nmol/L Bortezomib the HL60 cells' proliferation was significantly inhibited, the inhibition rate was 76%, and the cell nuclei became progressively pyknotic and were extensively fragmented. FCM showed apoptosis peaks 24 hours after treatment of Bortezomib of the concentrations of were 10 and 20 nmol/L, the apoptosis rate was 62.6%. Bcl-2 protein expression was down-regulated and the protein expressions of Caspase-9, Caspase-3, and PARP were all up-regulated.The mechanism of Bortezomib to induce apoptosis of myeloid leukemia cells is associated with down-regulation of Bcl-2 protein expression and cleaved activation of Caspase-9, Caspase-3 and PARP proteins.

Two SSR molecular markers, genomic-SSR and EST (expressed sequence tagged)-SSR, were used to measure the genetic diversity among 18 accessions of common wheat with known pedigrees, which were collected from winter wheat production region in Northern China. In addtion, the genetic diversity revealed by pedigree, EST-SSR and genomic-SSR was also compared. The results showed that the average number of alleles per genomic-SSR locus is 3.34, which is higher than that of EST-SSR (2.31), indicating that genomic-SSR is more polymorphic than EST-SSR. Genomic-SSR and EST-SSR were used to calculate the genetic distance (GD) in different materials. The mean GD value of EST-SSR for the 18 wheat genotypes is 0.3996,which is lower than that of genomic-SSR (0.5458). At the locus level, the mean GD calculated using pedigree information is higher (0. 9716) than that of genomic-SSR and EST-SSR markers (0.5458 and 0.3996). Therefore, although polymorphism of EST-SSR is low as compared to genomic SSR,it provides a more accurate evaluation of genetic relationship, especially when accessions are very closely related in pedigree. The strategy for improving the genetic diversity of wheat was also discussed.

In order to understand molecular basis of heterosis, mRNA differential display was used to analyze the differences in gene expression between seeds of 18 reciprocal hybrids and their 6 parents at 6th day after pollination. The relationship between gene expression patterns and heterosis was determined. Only bands that can be repeated in duplicate PCR were used for analysis so as to reduce false positive bands. Among the total of 2,025 bands displayed, 1,386 bands (68.43%) were reproducible. Eight patterns (fifteen kinds) of gene expression were observed, which include: (1) bands occurring in only one parent (two kinds); (2) bands observed in both hybrids and one parent (two kinds); (3) bands detected in parents and one hybrid (two kinds); (4) bands displayed in only one hybrid (two kinds); (5) bands revealed in one hybrid and its corresponding female (two kinds) or male (two kinds) parent; (6) bands visualized in only both hybrids (one kind); (7) bands occurring in only parents (one kind); (8) bands observed in parents and both hybrids (one kind). Our results indicate that differences of gene expression between hybrids and their parents are very obvious. The percentages of bands observed in only both hybrids and in only both parents are lower. The analysis shows that bands observed in parents and both hybrids are not correlated with all nine agronomic traits, which indicates differentially expressed genes are mainly responsible for the observed heterosis. At least one pattern of differential gene expression is significantly correlated with nine agronomic traits. Silenced bands in hybrid (which include bands occurring in only one parent, bands detected in both parents and one hybrid and bands in only parents) and bands present in one hybrid and its corresponding female or male parent are likely to play important roles in heterosis. These results suggest that early seed development could be closely related to heterosis.

The purpose of the present study was to observe whether primary afferent Abeta-fiber is involved in the information transmission between peripheral terminals of adjacent dermatomes. The dorsal cutaneous nerve branches of spinal nerves from T(8) to T(12) segments were cut proximally. One peripheral stump end of the cut nerves was dissected into a few filaments for the examination of mechanoreceptive properties of single Abeta-fibers and their discharges were observed while the other end was stimulated antidromically. Fifty Abeta-units were recorded in forty-two intact rats. After an electrical stimulation (0.45 mA, 0.1 ms, 20 Hz, for 10 s) was delivered to the stimulated nerve, the size of the receptive field of 60.6% (n=33) Abeta-fibers extended. The mean area of receptive fields of all examined units enlarged from 8.94+/-6.51 mm(2) to 20.34+/-16.17 mm(2) (P0.01) and the shapes of the receptive fields of 81.8% (n=20) units changed from a dot, round or ellipse with its long axis in parallel with the longitudinal axis of the body to an oblique ellipse with the longitudinal axis of the body. The mechanoreceptive threshold of 68.0% (n=50) units decreased with a reduction in mean threshold from 2.37+/-1.24 to 2.29+/-1.24 mN (P0.05). The duration of these changes in mechano-receptive properties increased from 52.23+/-9.27 to 56.93+/-15.76 min. Meanwhile, increasing discharge was found in 50.0% (n=50) units but lasted only for 1.52+/-0.46 min. The changes in mechanoreceptive properties appeared simultaneously with discharge changes but had longer duration than that of discharge change (P0.01). Discharges changes usually appeared in those units with the changes in mechanoreceptive properties following an antidromical electrical stimulation of adjacent spinal segment. These results suggest that low-threshold mechanoreceptive Abeta-fibers are affected by antidromical electrical stimulation of the cutaneous nerve from an adjacent spinal segment, indicating that information transmission occurs between the two endings of peripheral afferent nerves from adjacent spinal segments without any involvement of the central nervous system, and that Abeta-fibers are involved in the process of information transmission between peripheral terminals from adjacent spinal segments.

Simple sequence repeat (SSR) molecular marker was used to measure the genetic diversity of D-genome in 26 synthesized hexaploid wheat (AABBDD) introduced from CIMMYT. Twenty-three D-genome specific SSR primers were selected for PCR amplification, among which 22 primers can detect polymorphism. A total of 92 alleles were identified at 23 loci using the above SSR primers, with an average of 4 alleles per locus. The 92 alleles were used to calculate Nei's similarity index (GS) and the genetic distance (GD). It was also found that the mean genetic distance between 26 synthesized hexaploid wheat was 0.4955, which was obviously high. From the above results, it can be indicated that the genetic variation of D-genome in synthesized hexaploid wheat was abundant and could be used to improve the genetic diversity in wheat breeding. Interestingly, synthesized hexaploid wheat 17 and 18 shared the same D-genome donor, but three of 23 detected SSR loci were polymorphic between the two materials. Therefore, during the period of allopolyploidization, there was genetic differentiation in repeat region of donor genome.

It is important to estimate the genetic diversity between the parents for improving the heterosis of hybrid wheat. In this study, ISSR(inter-simple sequence repeat) marker was used to measure the genetic diversity within and among common wheat (Triticum aestivum L.), spelt (Triticum spelta L.), compactum (Triticum compactum Host.) and progeny of foreign wheat-based recurrent selection, and the possibility of establishing the new heterotic group was also assessed. Forty seven genotypes were used for ISSR analysis, which included 14 common wheat, 10 spelt wheat, 11 compactum and 12 progeny of recurrent selection. Eleven of 33 ISSR primers that can produce distinguishable bands were selected for PCR amplification. A total of 238 bands were amplified, among which 207 (87%) bands were polymorphic. The polymorphic bands amplified by each primer ranged from 11 to 38, with an averaged of 18.8. The percentage of polymorphic band (80.3%) in common wheat was higher than that in progeny of recurrent selection (78.7%), spelt (75.0%) and compactum (74.9%). The 238 polymorphic products were used to calculate Nei's similarity index (GS) and the genetic distance (GD). It was found that the mean genetic distance between different wheat types (0.3115-0.3442) was obviously higher than that within common wheat (0.2743), spelt (0.2351), compactum (0.2622). In addition, progeny of recurrent selection also showed much higher genetic distance with other three wheat types (0.3217, 0.3256, 0.3198). The cluster analysis was performed based on the genetic distance (GD) matrix by using UPGMA method. Common wheat, spelt, compactum and progeny of recurrent selection were classified into four different groups. In this study, ISSR marker was firstly used to assess genetic diversity among common wheat, spelt, compactum and progeny of recurrent selection, and can differentiate the wheat cultivars (lines) that selected from the same cross combination. It was concluded that spelt, compactum and progeny of recurrent selection can be used to diversify the genetic basis for hybrid wheat breeding and improve heterosis. It is possible to establish the wheat heterotic group by ISSR marker.

To investigate the effect of bortezomib alone or combined with harringtonine (HT) or arsenic trioxide (As2O3) on the proliferation activity and apoptosis of HL60/ADM cell line and refractory/relapse acute leukemia primary cell. HL60/ADM cells or refractory/relapse acute leukemia primary cells were incubated with bortezomib at different doses alone and in combination with HT or As2O3. The proliferation activity was abserved by MTT assay, cell apoptosis was studied by fluorescence microscopy and flow cytometry. Intracellular concentration of daunorubicin (DNR) was determined by flow cytometry. In bortezomib-treated tumor cells, inhibition rate enhanced and apoptotic cells increased in mumber time- and dose-dependently. 40nM Bortezomib can best inhibit the proliferation activity of HL60/ADM cells when incubated for 48 hours. 15uM As2O3 or 752nM HT combined with different doses of bortezomib can inhibit the proliferation and induce the apoptosis of HL60/ADM cells, The dual inhibition of As2O3 plus bortezomib or HT plus bortezomib may present a superior anticancer efficancy to either inhibition of the drugs above alone (P

Objective To explore the apoptosis effect induced by bortezomib combined with homoharringtonine or arsenious acid in HL-60 cell line and the mechanism.Methods Cell' s apoptosis was demonstrated by MTT assay and Hoechst33342 staining.Expression of bcl-2,Caspase-9,Caspase-3 and PARP protein was detected by Western blot.Results HL-60 cells' apoptosis could be induced by bortezomib,homoharringtonine and arsenious acid respectively.Proliferation inhibition of HL-60 cells could be enhanced significantly when treated by bortezomib combined with homoharringtonine or arsenious acid compared with treated by any of the three drugs alone (P ＜ 0.05).At the same time morphology shows the apoptosis induced by drugs combined is more obviously than by one drug.Western blot showed bcl-2 protein was down-regulated and Caspase-9,Caspase-3 and PARP proteins were all cleaved activation when cells were treated by 15 μmol/ L arsenious acid alone,but only cleaved activation of PARP and down-regulation of bcl-2 protein be detected when cells were treated with 30 nmol/L homoharringtonine alone,expression of Caspase-9 and Caspase-3 had no change compared with the control.The changes of associated proteins were paralleled with the cell' s apoptosis when treated with combined drugs.Conclusion HL-60 cells' apoptosis effect is inhanced significantly when bortezomib combined with homoharringtonine or arsenious acid.Arsenious acid and bortezomib can inhibit caspase signaling pathway and down-regulate the expression of bcl-2 protein together,but homoharringtonine and bortezomib can only down-regulate the expression of bcl-2 protein and induce cleaved activation of PARP together. Key words: Cells, HL-60; Bortezomib; Homoharringtonine; Arsenious acid; Apoptosis