[The Study of Recombinant Interfering Lentiviruses and Overexpressed Adenovirus Vectors Targeting Human c-Cbl Gene： Construction, Identification and Efficacy].

OBJECTIVE: To construct recombinant lentivirus and adenovirus which regulate the expression of c-Cbl gene and evaluate their efficacy. METHODS: The interference lentivirus and overexpressed adenovirus targeting human c-Cbl gene were constructed by gene recombination technology. Quantitative PCR and western blotting were used to detect the expression changes in c-Cbl gene and its transcription after leukemia cells (HL60,THP1) were infected by virus. RESULTS: Three recombinant interfering lentiviral vectors targeting human c-Cbl genes to successfully constructed and were identified by DNA sequencing, and the titers of the packaged viruses were all greater than 1×108 TU/ml. Among them, shRNA-2 lentivirus had the highest interference efficiency, and the expression of c-Cbl gene and CBL protein were decreased about 95% and 60% respectively after leukemia cells were infected with shRNA-2; In addition, the recombinant overexpression adenovirus targeting human c-Cbl gene was packaged successfully with the virus titer greater than 1×109 TU/ml. When leukemia cells were infected with adenovirus, the expression of c-Cbl gene and CBL protein were up-regulated about 10 times and 1.5 times respectively. CONCLUSION: Both recombinant interfering lentivirus and overexpression adenovirus can efficiently infect leukemia cells and affect the expressions of c-Cbl gene and CBL protein. It will lay a preliminary foundation for the subsequent study on the function of c-Cbl gene in tumor cells.

Distinct serotonergic pathways to the amygdala underlie separate behavioral features of anxiety.

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 (DRN5-HT∩vGluT3-BAPV) and pyramidal (DRN5-HT∩vGluT3-BAPyr) neurons have distinct intrinsic properties and gene expression and respond to anxiogenic and social stimuli, respectively. Activation of DRN5-HT∩vGluT3→BAPV inhibits 5-HT release via GABAB receptors on serotonergic terminals in BA, inducing social avoidance and avoidance of bright spaces. Activation of DRN5-HT∩vGluT3→BA neurons inhibits two subsets of BAPyr neurons via 5-HT1A receptors (HTR1A) and 5-HT1B receptors (HTR1B). Pharmacological inhibition of HTR1A and HTR1B in BA induces avoidance of bright spaces and social avoidance, respectively. These findings highlight the functional significance of heterogenic inputs from DRN to BA subpopulations in the regulation of separate anxiety-related behaviors.

Comparison of gene mutation spectra in younger and older Chinese acute myeloid leukemia patients and its prognostic value.

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.

Publisher Correction: Cannabinoid CB1 receptors in the amygdalar cholecystokinin glutamatergic afferents to nucleus accumbens modulate depressive-like behavior.

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.

Cannabinoid CB1 receptors in the amygdalar cholecystokinin glutamatergic afferents to nucleus accumbens modulate depressive-like behavior.

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.

High-density genetic linkage map construction and QTL mapping of grain shape and size in the wheat population Yanda1817 × Beinong6.

High-density genetic linkage maps are necessary for precisely mapping quantitative trait loci (QTLs) controlling grain shape and size in wheat. By applying the Infinium iSelect 9K SNP assay, we have constructed a high-density genetic linkage map with 269 F 8 recombinant inbred lines (RILs) developed between a Chinese cornerstone wheat breeding parental line Yanda1817 and a high-yielding line Beinong6. The map contains 2431 SNPs and 128 SSR & EST-SSR markers in a total coverage of 3213.2 cM with an average interval of 1.26 cM per marker. Eighty-eight QTLs for thousand-grain weight (TGW), grain length (GL), grain width (GW) and grain thickness (GT) were detected in nine ecological environments (Beijing, Shijiazhuang and Kaifeng) during five years between 2010-2014 by inclusive composite interval mapping (ICIM) (LOD ≥ 2.5). Among which, 17 QTLs for TGW were mapped on chromosomes 1A, 1B, 2A, 2B, 3A, 3B, 3D, 4A, 4D, 5A, 5B and 6B with phenotypic variations ranging from 2.62% to 12.08%. Four stable QTLs for TGW could be detected in five and seven environments, respectively. Thirty-two QTLs for GL were mapped on chromosomes 1B, 1D, 2A, 2B, 2D, 3B, 3D, 4A, 4B, 4D, 5A, 5B, 6B, 7A and 7B, with phenotypic variations ranging from 2.62% to 44.39%. QGl.cau-2A.2 can be detected in all the environments with the largest phenotypic variations, indicating that it is a major and stable QTL. For GW, 12 QTLs were identified with phenotypic variations range from 3.69% to 12.30%. We found 27 QTLs for GT with phenotypic variations ranged from 2.55% to 36.42%. In particular, QTL QGt.cau-5A.1 with phenotypic variations of 6.82-23.59% was detected in all the nine environments. Moreover, pleiotropic effects were detected for several QTL loci responsible for grain shape and size that could serve as target regions for fine mapping and marker assisted selection in wheat breeding programs.

A wheat lipid transfer protein 3 could enhance the basal thermotolerance and oxidative stress resistance of Arabidopsis.

Wheat (Triticum aestivum L.) is one of the major grain crops, and heat stress adversely affects wheat production in many regions of the world. Previously, we found a heat-responsive gene named Lipid Transfer Protein 3 (TaLTP3) in wheat. TaLTP3 was deduced to be regulated by cold, ABA, MeJA, Auxin and oxidative stress according to cis-acting motifs in its promoter sequences. In this study, we show that TaLTP3 is responsive to prolonged water deficit, salt or ABA treatment in wheat seedlings. Also, TaLTP3 accumulation was observed after the plant suffered from heat stress both at the seedling and the grain-filling stages. TaLTP3 protein was localized in the cell membrane and cytoplasm of tobacco epidermal cells. Overexpression of TaLTP3 in yeast imparted tolerance to heat stress compared to cells expressing the vector alone. Most importantly, transgenic Arabidopsis plants engineered to overexpress TaLTP3 showed higher thermotolerance than control plants at the seedling stage. Further investigation indicated that transgenic lines decreased H2O2 accumulation and membrane injury under heat stress. Taken together, our results demonstrate that TaLTP3 confers heat stress tolerance possibly through reactive oxygen species (ROS) scavenging.

Chronic administration of Liu Wei Dihuang protects rat's brain against D-galactose-induced impairment of cholinergic system.

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.

Screening and identification of the AFLP markers linked to a new powdery mildew resistance gene in wheat cultivar Brock.

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.

[Integrating genetic and gene expression data: methods and applications of eQTL mapping].

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.

[The effect of Haynaldia villosa v chromosome on the mitochondrial proteome of wheat-H. villosa chromosome substitution line and translocation line].

Wheat-Haynaldia villosa chromosome substitution line (6A/6V) and translocation lines (6DL/6VS, 6AL/6VS) were obtained through hybridization of H. villosa with powdery mildew susceptible cultivated wheat. Substitution line and translocation lines contain V chromosome or the chromosome short arm (VS) of H. villosa. They are resistant to powdery mildew. In this study, mitochondrial proteome changes were analyzed by using substitution line (6A/6V), translocation line (6DL/6VS) as experimental materials in order to studying the effects of V chromosome on the mitochondrial proteome and related to powdery mildew resistance. The results indicated that 16 new mitochondrial protein spots (spot1, 22kDa/PI8.5; spot2, 31 kDa/PI 7.5; spot3, 28 kDa/PI 7.0; spot4, 31 kDa/PI 6.5; spot5, 40 kDa/PI 7.5; spot6, 40 kDa/PI 7.4; spot7, 80 kDa/PI 8.4; spot8, 50 kDa/PI 7.5; spot9, 60 kDa/PI 7.3; spot10, 65 kDa/PI 6.6; spot11, 65 kDa/PI 6.6; spot12, 73 kDa/PI 7.5; spot13, 73 kDa/PI 7.7; spot14, 46 kDa/PI 7.4; spot15, 46 kDa/PI 7.3; spot16, 38 kDa/PI 6.3) were produced and 7 mitochondrial protein spots (spot1, 40 kDa/PI 7.5; spot2, 43 kDa/PI 7.6; spot3, 48 kDa/PI 7.5; spot4, 42 kDa/PI 8.0; spot5, 43 kDa/PI 7.5; spot6, 32 kDa/PI 4.8; spot7, 40 kDa/PI 5.5) were absent in substitution line, 7 new mitochondrial protein spots (spotl, 43 kDa/PI 6.3; spot2, 60 kDa/PI 6.5; spot3, 60 kDa/PI 6.4; spot4, 65 kDa/PI 7.5; spot5, 55 kDa/PI 8.2; spot6, 31 kDa/PI 8.0; spot7, 43 kDa/PI 8.0) were produced and 6 mitochondrial protein spots (spot1', 66 kDa/PI 8.3; spot2', 58 kDa/PI 8.5; spot3', 36 kDa/PI 7.0; spot4', 48 kDa/PI 7.7; spot5', 48 kDa/PI 6.8; spot6', 43 kDa/PI 6.2) were absent in translocation line. These experimental results suggest that V chromosome or VS of H. villosa can obviously lead mitochondrial proteome changed. These changes may be associated with resistant to powdery mildew of substitution line and translocation line.

Paracrine-like excitation of low-threshold mechanoceptive C-fibers innervating rat hairy skin is mediated by substance P via NK-1 receptors.

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').

[In vitro effect of bortezomib alone or in combination with harringtonine or arsenic trioxide on proliferation and apoptosis of multidrug resistant leukemia cells].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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 (P < 0.05, P < 0.01). Bortezomib (10 nmol/L) could markedly enhance the intracellular accumulation of DNR in HL-60/ADM cells (P < 0.05). CONCLUSIONS: 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.

[STI571 enhances the effect of arsenic trioxide and Velcade on bcr/abl+-CD34+ cell proliferation and apoptosis].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: Combination with STI571 enhances the effects of As2O3 and Velcade on bcr/abl+-CD34+ cells, suggesting the potential clinical value of this regimen.

[Effect of bortezomib used alone or in combination with arsenic trioxide on HL-60 cell xenograft in nude mice].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[HL-60 cell apoptosis induced by bortezomib alone or in combination with arsenic trioxide in vitro].

OBJECTIVE: To investigate the effect of bortezomib alone and in combination with arsenic trioxide on apoptosis of HL-60 cells. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Induction of apoptosis in HL-60 cells by bortezomib alone or in combination with harringtonine in vitro].

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.

[The changes in chloroplast proteome caused by different nuclear background in cytoplasmic male-sterile (CMS) of wheat].

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.

[Effect of G-CSF on the proliferation and differentiation of bcr/abl(+)-CD34+ cells from CML patients].

OBJECTIVE: To study the effect of granulocyte colony-stimulating factor (G-CSF) on the proliferation and differentiation of bcr/abl(+)-CD34+ cells. METHODS: 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. RESULTS: 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 (P < 0.05) , 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 (P < 0.05, P < 0.01, P < 0.01, 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 (P < 0. 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. CONCLUSIONS: 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.

[The mixed major gene plus polygenes inheritance for female fertility in wheat (Triticum aestivum L.)].

Three sets of data for the P1, P2, F1, and F2 populations derived from three crosses between the normal fertility wheat (Triticum aestivum L.) cultivars with different ecotypes and the female sterile line (XND126) were used to investigate the inheritance of female fertility in wheat using mixed major gene plus polygenes inheritance model in 2005 and 2006. The results from the joint segregation analysis of the four generations showed that female fertility in wheat is controlled by two major genes plus polygenes, and the interaction between the two major genes is also detected.