Selectivity Quantification with the Fluorescent Quantitative Model-Assisted Semi-Selective Probe (Carbon Dots): Accurate Determination of Chlortetracycline in Aqueous Environments with Interference.

Probes such as carbon dots (C-dots) have extensive and important applications in the quantitative analysis of complex biological and environmental systems. However, the development of probes is often hindered by incomplete selectivity, i.e., a probe that responds to one substance is also prone to respond to coexisting structurally similar substances. Therefore, the above dilemma often leads to be developed as semi-selective probes, so that the development of probes is abandoned halfway. This work shows how a semi-selective probe can enhance selectivity by combining a proper multivariate calibration model. Primarily, we developed a semi-selective fluorescent probe that responded to tetracyclines (TCs) with discarded tobacco leaves. Then, we introduced the multivariate quantitative fluorescence model (QFM) to enhance its selectivity and solve the problem of fluorescence spectral shift. For the determination of chlortetracycline (CTC) with this semi-selective C-dots probe in mineral and lake water samples and compared to the traditional quantitative model, the introduced QFM resulted in an average relative predictive error (ARPE) in mineral water spiked samples decreased from 57.1 to 5.6%, which reduced the ARPE in the lake water spiked samples from 18.1 to 4.7%. The above results show that the QFM-assisted semi-selective probe C-dots strategy (QFMC-dots) can enhance selectivity, and QFMC-dots achieved high-selective and accurate determination of CTC in interfering mineral and lake water samples, with the limit of detection and limit of quantitation of 0.55 and 1.66 µM, respectively. The proposed strategy of enhancing selectivity by introducing a proper multivariate calibration model can reduce the difficulty and increase success rate of developing probes, which can be expected to provide an interesting alternative for the development of probes, especially when encountering semi-selective problems.

Interference-free quantitation of aromatic amino acids in two complex systems by three-way calibration with ultraviolet-visible spectrophotometer: Exploration of trilinear decomposition of spectrum-pH data.

Aromatic amino acids play an extremely important role in life activities and participate in many biological processes. Their concentration levels are associated with a variety of diseases, such as phenylketonuria and colorectal cancer. Therefore, the quantification of aromatic amino acids is an important task. In the present work, a novel and rapid three-way analytical method was proposed to detect the levels of aromatic amino acids in prostate cancer cells (PC3 cells) and Dulbecco's modified minimal essential medium (DMEM cell culture), by using the affordable ultraviolet-visible spectrophotometer. First, spectrum-pH second-order data were designed per sample; Second, properties of the resulted spectrum-pH-sample three-way data were investigated by utilizing the parallel factor analysis (PARAFAC), alternating trilinear decomposition (ATLD), and constrained alternating trilinear decomposition (CATLD) algorithms, and a flexible scanning approach for determining the constraint parameters of CATLD was proposed; Third, a three-way calibration method based on the CATLD algorithm with the proposed scanning approach was developed for interference-free quantification of aromatic amino acids in these systems. The average relative predictive errors of validation (ARPEV) for phenylalanine, tyrosine, and tryptophan were 1.4%, 3.0%, and 0.7% in prostate cancer cells, and ARPEV for phenylalanine, tyrosine, and tryptophan were 4.1%, 1.2%, and 0.7% in DMEM cell culture. The predicted contents of tyrosine and tryptophan in DMEM cell culture were 64.2 ± 2.9 µg mL-1, 5.6 ± 0.3 µg mL-1, there are no significant differences in the concentrations between the developed analytical method and high performance liquid chromatography method. The proposed spectrum-pH-sample three-way calibration method based on CATLD algorithm can provide an interesting analytical strategy with high selectivity and accuracy for ultraviolet-visible spectrophotometer.

The cytokine profiles in follicular fluid and reproductive outcomes in women with endometriosis.

PROBLEM: Endometriosis patients undergoing in vitro fertilization-embryo transfer (IVF-ET) treatment suffer from poor oocyte quality, a reduced likelihood of the fertilization rate, and low embryo quality. The dysregulation of immune cells and cytokine profiles in the follicular fluid (FF) may play an important role in the competence of the oocyte and the development of the embryo, but the mechanism remains largely unknown. METHOD OF STUDY: A total of 40 proved advanced staged endometriosis patients were enrolled in this study. The pregnancy results were followed until all the embryos collected by the first oocyte retrieval cycle were used up. The immune cells subtypes in FF and serum collected on the day of oocyte retrieval were detected by flow cytometry and 27 cytokines were determined using the Bio-Plex Pro Human Cytokine 27-Plex Immunoassay. The specific effect of cytokine on the gene expression of human granulosa cells was determined by RT-qPCR. RESULTS: The fertilization rate and the cumulative live birth rate were significantly lower in the endometriosis group. The ratio of CD4+ /CD8+ T cells in FF was significantly lower, while the level of IP-10, RANTES and G-CSF were statistically higher in the endometriosis group. The level of IP-10 correlated with the IVF outcome. Moreover, treated by IP-10, the mRNA level of FSHR and CYP19A1 the human granulosa cells were downregulated in vitro. CONCLUSION: These results suggest that alterations of the lymphocyte subsets and cytokines in women with advanced endometriosis may have an impact on the oocyte development and resulting in poorer IVF outcomes.

The Role of Traditional Chinese Formula Ding-Kun Pill (DKP) in Expected Poor Ovarian Response Women (POSEIDON Group 4) Undergoing In Vitro Fertilization-Embryo Transfer: A Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial.

Objective: The primary objective of the study was to assess traditional Chinese formula DKP supplementation in terms of efficacy and safety on reproductive outcomes of expected poor ovarian responder (POR, POSEIDON Group 4) undergoing in vitro fertilization-embryo transfer (IVF-ET). Design Setting and Participants: Women eligible for IVF-ET were invited to participate in this randomized, double-blind, placebo-controlled, superiority trial at academic fertility centers of ten public hospitals in Chinese Mainland. A total of 462 patients (35-44 years) equally divided between DKP and placebo groups with antral follicle count (AFC) <5 or anti-müllerian hormone (AMH) <1.2 ng/ml were randomized. Interventions: All participants were given DKP or 7 g placebo twice daily on the previous menstrual cycle day 5 until oocyte retrieval, which took approximately 5 to 6 weeks. Main Outcome Measure: The primary outcome was the ongoing pregnancy defined as more than 20 gestational weeks of an intrauterine living fetus confirmed by pelvic ultrasonography. Results: Demographic characteristics were equally distributed between the study populations. Intention-to-treat (ITT) analysis revealed that ongoing pregnancy rate (OPR) was not significantly different between DKP and placebo groups [26.4% (61/231) versus 24.2% (56/231); relative risk (RR) 1.09, 95% confidence interval (CI) 0.80 to 1.49, P = 0.593]. No significant differences between groups were observed for the secondary outcomes. The additional per protocol (PP) analysis was in line with ITT results: OPR in DKP group was 27.2% (61/224) versus 24.1% (55/228) in placebo group [RR 1.13, 95%CI (0.82 to 1.55), P = 0.449]. After subgroup analysis the findings concluded that POR population of 35-37 years had a significantly higher OPR after 5-6 weeks of oral DKP (41.8%, 33/79) versus placebo (25.4%, 18/71) [RR 1.65, 95% CI (1.02 to 2.65), P = 0.034, P for interaction = 0.028]. Conclusion: This well-designed randomized controlled trial (RCT) offers new high-quality evidence to supplement existing retrospective literature concerning DKP performance in expected PORs. DKP could be recommended as a safe and natural remedy for expected PORs (aged 35-37 years) who fulfill the POSEIDON group 4 criteria. However, additional interventional clinical studies are undoubtedly required to be conducted in the future to validate this hypothesis. Clinical Trial Registration: www.chictr.org.cn, identifier ChiCTR1900026614.

The Cotton Apoplastic Protein CRR1 Stabilizes Chitinase 28 to Facilitate Defense against the Fungal Pathogen Verticillium dahliae.

The apoplast serves as the first battlefield between the plant hosts and invading microbes; therefore, work on plant-pathogen interactions has increasingly focused on apoplastic immunity. In this study, we identified three proteins in the apoplast of cotton (Gossypium sp) root cells during interaction of the plant with the fungal pathogen Verticillium dahliae Among these proteins, cotton host cells secrete chitinase 28 (Chi28) and the Cys-rich repeat protein 1 (CRR1), while the pathogen releases the protease VdSSEP1. Biochemical analysis demonstrated that VdSSEP1 hydrolyzed Chi28, but CRR1 protected Chi28 from cleavage by Verticillium dahliae secretory Ser protease 1 (VdSSEP1). In accordance with the in vitro results, CRR1 interacted with Chi28 in yeast and plant cells and attenuated the observed decrease in Chi28 level that occurred in the apoplast of plant cells upon pathogen attack. Knockdown of CRR1 or Chi28 in cotton plants resulted in higher susceptibility to V. dahliae infection, and overexpression of CRR1 increased plant resistance to V dahliae, the fungus Botrytis cinerea, and the oomycete Phytophthora parasitica var nicotianae By contrast, knockout of VdSSEP1 in V. dahliae destroyed the pathogenicity of this fungus. Together, our results provide compelling evidence for a multilayered interplay of factors in cotton apoplastic immunity.

iTRAQ-based proteomics analysis of autophagy-mediated immune responses against the vascular fungal pathogen Verticillium dahliae in Arabidopsis.

The mechanisms underlying the functional link between autophagy and plant innate immunity remain largely unknown. In this study, we investigated the autophagy-mediated plant defense responses against Verticillium dahliae (V. dahliae) infection by comparative proteomics and cellular analyses. An assessment of the autophagy activity and disease development showed that autophagic processes were tightly related to the tolerance of Arabidopsis plant to Verticillium wilt. An isobaric tags for relative and absolute quantification (iTRAQ)-based proteomics analysis was performed, and we identified a total of 780 differentially accumulated proteins (DAPs) between wild-type and mutant atg10-1 Arabidopsis plants upon V. dahliae infection, of which, 193 ATG8-family-interacting proteins were identified in silico and their associations with autophagy were verified for several selected proteins. Three important aspects of autophagy-mediated defense against V. dahliae infection were revealed: 1) autophagy is required for the activation of upstream defense responses; 2) autophagy-mediated mitochondrial degradation (mitophagy) occurs and is an important player in the defense process; and 3) autophagy promotes the transdifferentiation of perivascular cells and the formation of xylem hyperplasia, which are crucial for protection against this vascular disease. Together, our results provide several novel insights for understanding the functional association between autophagy and plant immune responses.

iTRAQ-based proteomic analysis of defence responses triggered by the necrotrophic pathogen Rhizoctonia solani in cotton.

The soil-borne necrotrophic pathogen fungus Rhizoctonia solani is destructive, causing disease in various important crops. To date, little is known about the host defence mechanism in response to invasion of R. solani. Here, an iTRAQ-based proteomic analysis was employed to investigate pathogen-responsive proteins in the disease tolerant/resistant cotton cultivar CRI35. A total of 174 differentially accumulated proteins (DAPs) were identified after inoculation of cotton plants with R. solani. Functional categorization analysis indicated that these DAPs can be divided into 12 subclasses. Notably, a large portion of DAPs are known to function in reactive oxygen species (ROS) metabolism and the expression of several histone-modifying and DNA methylating proteins were significantly induced upon challenge with the fungus, indicating that the redox homeostasis and epigenetic regulation are important for cotton defence against the pathogen. Additionally, the expression of proteins involved in phenylpropanoid biosynthesis was markedly changed in response to pathogen invasion, which may reflect a particular contribution of secondary metabolism in protection against the fungal attack in cotton. Together, our results indicate that the defence response of cotton plants to R. solani infection is active and multifaceted and involves the induction of proteins from various innate immunity-related pathways. SIGNIFICANCE: Cotton damping-off is a destructive disease caused by the necrotrophic fungus Rhizoctonia solani. To date, the host defence mechanism involved in the disease protection remains largely unknown. Here, we reported the first proteomic analysis on cotton immune responses against R. solani infection. Employing iTRAQ technique, we obtained a total of 174 differentially accumulated proteins (DAPs) that can be classified into 12 functional groups. Further analysis indicated that ROS homeostasis, epigenetic regulation and phenylpropanoid biosynthesis were tightly associated with the innate immune responses against R. solani infection in cotton. The obtained data provide not only important information for understanding the molecular mechanism involved in plant-R. solani interaction but also application clues for genetic breeding of crops with improved R. solani resistance.

The cotton MYB108 forms a positive feedback regulation loop with CML11 and participates in the defense response against Verticillium dahliae infection.

Accumulating evidence indicates that plant MYB transcription factors participate in defense against pathogen attack, but their regulatory targets and related signaling processes remain largely unknown. Here, we identified a defense-related MYB gene (GhMYB108) from upland cotton (Gossypium hirsutum) and characterized its functional mechanism. Expression of GhMYB108 in cotton plants was induced by Verticillium dahliae infection and responded to the application of defense signaling molecules, including salicylic acid, jasmonic acid, and ethylene. Knockdown of GhMYB108 expression led to increased susceptibility of cotton plants to V. dahliae, while ecotopic overexpression of GhMYB108 in Arabidopsis thaliana conferred enhanced tolerance to the pathogen. Further analysis demonstrated that GhMYB108 interacted with the calmodulin-like protein GhCML11, and the two proteins form a positive feedback loop to enhance the transcription of GhCML11 in a calcium-dependent manner. Verticillium dahliae infection stimulated Ca(2+) influx into the cytosol in cotton root cells, but this response was disrupted in both GhCML11-silenced plants and GhMYB108-silenced plants in which expression of several calcium signaling-related genes was down-regulated. Taken together, these results indicate that GhMYB108 acts as a positive regulator in defense against V. dahliae infection by interacting with GhCML11. Furthermore, the data also revealed the important roles and synergetic regulation of MYB transcription factor, Ca(2+), and calmodulin in plant immune responses.

Cotton major latex protein 28 functions as a positive regulator of the ethylene responsive factor 6 in defense against Verticillium dahliae.

In this study, we identified a defense-related major latex protein (MLP) from upland cotton (designated GhMLP28) and investigated its functional mechanism. GhMLP28 transcripts were ubiquitously present in cotton plants, with higher accumulation in the root. Expression of the GhMLP28 gene was induced by Verticillium dahliae inoculation and was responsive to defense signaling molecules, including ethylene, jasmonic acid, and salicylic acid. Knockdown of GhMLP28 expression by virus-induced gene silencing resulted in increased susceptibility of cotton plants to V. dahliae infection, while ectopic overexpression of GhMLP28 in tobacco improved the disease tolerance of the transgenic plants. Further analysis revealed that GhMLP28 interacted with cotton ethylene response factor 6 (GhERF6) and facilitated the binding of GhERF6 to GCC-box element. Transient expression assay demonstrated that GhMLP28 enhanced the transcription factor activity of GhERF6, which led to the augmented expression of some GCC-box genes. GhMLP28 proteins were located in both the nucleus and cytoplasm and their nuclear distribution was dependent on the presence of GhERF6. Collectively, these results demonstrate that GhMLP28 acts as a positive regulator of GhERF6, and synergetic actions of the two proteins may contribute substantially to protection against V. dahliae infection in cotton plants.

Inhibition of reactive oxygen species in hypothalamic paraventricular nucleus attenuates the renin-angiotensin system and proinflammatory cytokines in hypertension.

AIMS: To explore whether reactive oxygen species (ROS) scavenger (tempol) in the hypothalamic paraventricular nucleus (PVN) attenuates renin-angiotensin system (RAS) and proinflammatory cytokines (PICs), and decreases the blood pressure and sympathetic activity in angiotensin II (ANG II)-induced hypertension. METHODS AND RESULTS: Male Sprague-Dawley rats were infused intravenously with ANG II (10 ng/kg per min) or normal saline (NS) for 4 weeks. These rats were treated with bilateral PVN infusion of oxygen free radical scavenger tempol (TEMP, 20 µg/h) or vehicle (artificial cerebrospinal fluid, aCSF) for 4 weeks. ANG II infusion resulted in increased mean arterial pressure (MAP) and renal sympathetic nerve activity (RSNA). These ANG II-infused rats also had higher levels of gp91(phox) (a subunit of NAD(P)H oxidase), angiotensin-converting enzyme (ACE), and interleukin-1 beta (IL-1ß) in the PVN than the control animals. Treatment with PVN infusion of TEMP attenuated the overexpression of gp91(phox), ACE and IL-1ß within the PVN, and decreased sympathetic activity and MAP in ANG II-infused rats. CONCLUSION: These findings suggest that ANG II infusion induces elevated PICs and oxidative stress in the PVN, which contribute to the sympathoexcitation in hypertension. Inhibition of reactive oxygen species in hypothalamic paraventricular nucleus attenuates the renin-angiotensin system, proinflammatory cytokines and oxidative stress in ANG II-induced hypertension.

Exercise training attenuates hypertension and cardiac hypertrophy by modulating neurotransmitters and cytokines in hypothalamic paraventricular nucleus.

AIMS: Regular exercise as an effective non-pharmacological antihypertensive therapy is beneficial for prevention and control of hypertension, but the central mechanisms are unclear. In this study, we hypothesized that chronic exercise training (ExT) delays the progression of hypertension and attenuates cardiac hypertrophy by up-regulating anti-inflammatory cytokines, reducing pro-inflammatory cytokines (PICs) and restoring the neurotransmitters balance in the hypothalamic paraventricular nucleus (PVN) in young spontaneously hypertensive rats (SHR). In addition, we also investigated the involvement of nuclear factor-κB (NF-κB) p65 and NAD(P)H oxidase in exercise-induced effects. METHODS AND RESULTS: Moderate-intensity ExT was administrated to young normotensive Wistar-Kyoto (WKY) and SHR rats for 16 weeks. SHR rats had a significant increase in mean arterial pressure and cardiac hypertrophy. SHR rats also had higher levels of glutamate, norepinephrine (NE), phosphorylated IKKß, NF-κB p65 activity, NAD(P)H oxidase subunit gp91(phox), PICs and the monocyte chemokine protein-1 (MCP-1), and lower levels of gamma-aminobutyric acid (GABA) and interleukin-10 (IL-10) in the PVN. These SHR rats also exhibited higher renal sympathetic nerve activity (RSNA), and higher plasma levels of PICs, and lower plasma IL-10. However, ExT ameliorates all these changes in SHR rats. CONCLUSION: These findings suggest that there are the imbalances between excitatory and inhibitory neurotransmitters and between pro- and anti-inflammatory cytokines in the PVN of SHR rats, which at least partly contributing to sympathoexcitation, hypertension and cardiac hypertrophy; chronic exercise training attenuates hypertension and cardiac hypertrophy by restoring the balances between excitatory and inhibitory neurotransmitters and between pro- and anti-inflammatory cytokines in the PVN; NF-κB and oxidative stress in the PVN may be involved in these exercise-induced effects.

Identification of the proteins in green cotton fiber using a proteomics-based approach.

The proteome of 21 DPA (days post-anthesis) of green cotton fiber was analyzed using two-dimensional gel electrophoresis to yield the first reference proteome map. Of 220 individual spots that were excised and analyzed by MALDI-TOF/TOF MS, 156 were identified and cataloged according to their functions. Twelve classes of proteins were identified. Many of these proteins were related to carbohydrate metabolism and energy production. Other notable functional classes included oxidoreductase, cell wall-related and cytoskeleton proteins. This study gives a snapshot of the proteome in 21 DPA green cotton fiber and advances our understanding of molecular mechanisms related to pigment biosynthesis in green cotton fiber.

A comparative proteomic analysis provides insights into pigment biosynthesis in brown color fiber.

A comparative proteomic analysis was performed to identify the differences between brown cotton fiber and a white near-isogenic line, and 78 differential spots were identified at three time points (12-, 18-, and 24-day post-anthesis [DPA]) using MALDI-TOF/TOF. Our data illustrate several aspects of pigment synthesis and fiber development in brown color fiber (BCF). First, 21 spots were associated with secondary metabolism; 15 of these with high abundance in BCF were involved in flavonoid biosynthesis. Second, several spots with lower abundance in BCF were found. Thirteen spots were related to energy/carbohydrate metabolism; in particular, spots related to the glycolytic pathway exhibited lower abundance in 12 DPA BCF. Several spots related to redox homeostasis, cytoskeleton, and protein metabolism also showed lower abundance in BCF, including proteins that are critical for fiber development, such as ascorbate peroxidase, superoxide dismutase, actin, annexin and heat shock protein. Third, several proteins such as leucine aminopeptidase preprotein and progesterone-5-beta-reductase were newly identified proteins in cotton fibers. These findings demonstrated the presence of a complicated metabolic network in BCF and advanced our understanding of the molecular mechanisms of pigment biosynthesis in colored cotton, which will provide new insight for the development of new color types by genetic manipulation.

[Cloning and functional analysis of chitinase gene GbCHI from sea-island cotton (Gossypium barbadense)].

Chitinase is one of the important pathogenesis-related (PR) proteins in plants. By comparative proteomics study, a novel pathogen-responsive chitinase (known as GbCHI) has been identified from sea-island cotton (Gossypium barbadense). The GbCHI cDNA was cloned from wilt-resistant sea-island cotton and the anti-fungal activity of the gene product was investigated. qRT-PCR analysis indicated that GbCHI was expressed constitutively in root, stem, leaf, flower, and ovule of cotton plant, and the expression could be induced by Verticillium dahliae and plant hormone SA, ACC, and JA. Subcellular localization analysis using GFP-tagged proteins showed that GbCHI-GFP fusion proteins were targeted mainly to the plasma membrane. Anti-fungal assay demonstrated that GbCHI could inhibit spore germination and hyphae growth of V. dahliae significantly. These results provide important information for understanding the cellular function of GbCHI and for exploring the application potential of this gene in molecular breeding of wilt-tolerant cotton plants.

(1)H and (13)C NMR Assignments of Cytotoxic 3S-1,2,3,4-Tetrahydro-ß-carboline-3-carboxylic Acid from the Leaves of Cichorium endivia.

An amino acid, 3S-1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acid, was isolated for the first time from the leaves of Cichorium endivia. The complete assignment of its (1)H and (13)C NMR spectroscopic data was carried out also for the first time based on extensive 1D and 2D NMR experiments. Cytotoxicity of this isolated compound against HCT-8 and HepG2 human cancer cell lines was evaluated for the first time, with moderate activities being found.

(3S)-1,2,3,4-Tetrahydro-ß-carboline-3-carboxylic acid from Cichorium endivia. L induces apoptosis of human colorectal cancer HCT-8 cells.

Cichorium endivia. L, consumed either cooked or eaten raw in salads, is a popular kind of vegetable cultivated all around the World. Its components have been widely used in folk medicine in anti-inflammatory therapy. However, the anti-cancer activity of the components has never been reported. In this study, (3S)-1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acid (1), an amino acid isolated from C. endivia. L, was found for the first time to show cytotoxic activity in colorectal cancer cell line HCT-8. Compound 1 at concentrations of 0.5-4 µM induced apoptosis of HCT-8 cells in a dose-dependent manner. The compound 1-induced apoptosis in HCT-8 cells was accompanied by the loss of mitochondrial membrane potential, the activation of caspase-3, caspase-8 and caspase-9, the up-regulation of Bax and the down-regulation of Bcl-2. In addition, compound 1 suppressed the activation of NF-κB, which acts as an inhibitor of apoptosis. Taken together, these results suggested that compound 1 could significantly induce apoptosis of HCT-8 cells through the suppression of NF-κB signaling pathway, and thus can be considered as a potential candidate for developing chemotherapeutic drugs against cancer.

[Analysis of sex chromosome mosaicisms in early cleavage-stage human embryos and blastocysts with poor embryo quality scores].

OBJECTIVE: To analyze sex chromosome mosaicisms in early cleavage-stage human embryos and blastocysts with poor embryo quality score based on the numbers of pronucleus(PN) zygotes using X,Y dual color fluorescence in situ hybridization (FISH), and to discuss the possible mechanisms. METHODS: Fresh or frozen-thawed early cleavage-stage human embryos and blastocysts with poor embryo quality score not suitable for embryo transfer were studied with dual color FISH. RESULTS: Double signal rate of 2PN among early cleavage-stage embryos was 66.67%, which was significantly higher than 1PN and 3PN embryos. Single signal rate of 1PN early cleavage-stage embryos was 90.41%, which was significantly higher than 2PN and 3PN ones. Three signal rate of 3PN early cleavage-stage embryos was 28.00%, which was significantly higher than 1PN and 2PN ones. Double signal rate of 3PN ones was 46.00%, which was significantly higher than 1PN ones. The polyploid rate of frozen-thawed early cleavage-stage embryos was 23.53%, which was slightly higher than that of fresh embryos, but with no statistical significance. The mosaicism rate of 24 blastocysts was 100.00% and the double signal dominant (≥ 50%) rate was 62.50%, which was significantly higher than the rate of early cleavage-stage embryos. CONCLUSION: Using 2PN as the criterion for embryo quality score cannot guarantee the selection of normal fertilized embryo for transplantation. Frozen-thawed embryos may harbor more polyploid cells. To avoid the selection of embryos with abnormal chromosomes, combinations of pre-implantation genetic diagnosis (PGD) and prenatal diagnosis are necessary. Meanwhile, blastocysts with poor quality scores may provide an important source for embryo stem cells.

Proteomic analysis of the sea-island cotton roots infected by wilt pathogen Verticillium dahliae.

Verticillium wilt of cotton is a vascular disease mainly caused by the soil-born filamentous fungus Verticillium dahliae. To study the mechanisms associated with defense responses in wilt-resistant sea-island cotton (Gossypium barbadense) upon V. dahliae infection, a comparative proteomic analysis between infected and mock-inoculated roots of G. barbadense var. Hai 7124 (a cultivar showing resistance against V. dahliae) was performed by 2-DE combined with local EST database-assisted PMF and MS/MS analysis. A total of 51 upregulated and 17 downregulated proteins were identified, and these proteins are mainly involved in defense and stress responses, primary and secondary metabolisms, lipid transport, and cytoskeleton organization. Three novel clues regarding wilt resistance of G. barbadense are gained from this study. First, ethylene signaling was significantly activated in the cotton roots attacked by V. dahliae as shown by the elevated expression of ethylene biosynthesis and signaling components. Second, the Bet v 1 family proteins may play an important role in the defense reaction against Verticillium wilt. Third, wilt resistance may implicate the redirection of carbohydrate flux from glycolysis to pentose phosphate pathway (PPP). To our knowledge, this study is the first root proteomic analysis on cotton wilt resistance and provides important insights for establishing strategies to control this disease.