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Morphological novelties, including formation of trait combinations, may result from de novo gene origination and/or co-option of existing genes into other developmental contexts. A variety of shape-color combinations of capitular florets occur in Chrysanthemum and its allies. We hypothesized that co-option of a carotenoid cleavage dioxygenase gene into the floral symmetry gene network would generate a white zygomorphic ray floret. We tested this hypothesis in an evolutionary context using species in Chrysanthemum sensu lato, a monophyletic group with diverse floral shape-color combinations, based on morphological investigation, interspecific crossing, molecular interaction and transgenic experiments. Our results showed that white color was significantly associated with floret zygomorphy. Specific expression of the carotenoid cleavage dioxygenase gene CCD4a in marginal florets resulted in white color. Crossing experiments between Chrysanthemum lavandulifolium and Ajania pacifica indicated that expression of CCD4a is trans-regulated. The floral symmetry regulator CYC2g can activate expression of CCD4a with a dependence on TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING (TCP) binding element 8 on the CCD4a promoter. Based on all experimental findings, we propose that gene co-option of carotenoid degradation into floral symmetry regulation, and the subsequent dysfunction or loss of either CYC2g or CCD4a, may have led to evolution of capitular shape-color patterning in Chrysanthemum sensu lato.

Renal cell carcinoma (RCC) has the highest mortality rate among urological cancers and tumor angiogenesis that plays a critical role in RCC progress. Epidermal growth factor-like domain multiple 7 (EGFL7) has been recently identified as a regulator in RCC tumor angiogenesis and progression. Long noncoding RNA (LncRNA) HOTAIR has been considered as a pro-oncogene in multiple cancers, but its precise mechanism of tumor angiogenesis has rarely been reported. MicroRNA-126 (miR-126) functions as a tumor suppressor in RCC. However, the underlying tumor angiogenesis mechanism of HOTAIR/miR-126 axis in RCC has not been studied. The proliferation, migration, angiogenesis, and expression of EGFL7 and related proteins in extracellular signal-regulated kinase (ERK)/activators of transcription 3 (STAT3) signal pathway were determined to examine the effect and mechanism of HOTAIR and miR-126 on RCC progress. The regulatory relationship of HOTAIR and miR-126, as well as miR-126 and EGFL7 were tested using dual-luciferase reporter assay. Aenograft RCC mice model was used to examine the effect of HOTAIR on RCC tumor growth and metastasis in vivo. HOTAIR knockdown and miR-126 overexpression suppressed the proliferation, migration, and angiogenesis of RCC cells. HOTAIR regulated EGFL7 expression by competitively binding to miR-126. Knockdown of HOTAIR significantly suppressed the RCC tumor progression and lung metastasis in vivo. These findings suggest that lncRNA HOTAIR regulate RCC angiogenesis through miR-126/EGFL7 axis and provide a new perspective on the molecular pathways of angiogenesis in RCC development, which might be potential therapeutic targets for RCC treatment.

Jiarui Zhang, Rui Zhao, Dongliang Xing, Jing Cao, Yan Guo, Liang Li, Yanan Sun, Linli Tian, Ming Liu Department of Otorhinolaryngology, Head and Neck Surgery, The Second Affiliated Hospital, Harbin Medical University, Harbin City, Heilongjiang Province 150086, People’s Republic of ChinaCorrespondence: Linli Tian; Ming LiuDepartment of Otorhinolaryngology, Head and Neck Surgery, The Second Affiliated Hospital, Harbin Medical University, No. 148 Baojian Road, Nangang District, Harbin City, Heilongjiang Province 150086, People’s Republic of ChinaEmail zt0675@163.com; qr9830@163.comBackground: Magnesium isoglycyrrhizinate (MI) was extracted from roots of the plant Glycyrrhiza glabra, which displays multiple pharmacological activities such as anti-inflammation, anti-apoptosis, and anti-tumor. Here, we aimed to investigate the effect of MI on the progression and epithelial–mesenchymal transition (EMT) of laryngeal cancer.Methods: Forty laryngeal cancer clinical samples were used. The role of MI in the proliferation of laryngeal cancer cells was assessed by MTT assay, Edu assay and colony formation assay. The function of MI in the migration and invasion of laryngeal cancer cells was tested by transwell assays. The effect of MI on apoptosis of laryngeal cancer cells was determined by cell apoptosis assay. The impact of MI on tumor growth in vivo was analyzed by tumorigenicity analysis using Balb/c nude mice. qPCR and Western blot analysis were performed to measure the expression levels of gene and protein, respectively.Results: We identified that EMT-related transcription factor Twist was significantly elevated in the laryngeal cancer tissues. The expression of Twist was also enhanced in the human laryngeal carcinoma HEP-2 cells compared with that in the primary laryngeal epithelial cells. The high expression of Twist was remarkably correlated with poor overall survival of patients with laryngeal cancer. Meanwhile, our data revealed that MI reduced cell proliferation, migration and invasion and enhanced apoptosis of laryngeal cancer cells in vitro. Moreover, MI decreased transcriptional activation and the expression levels of NF-κB and Twist, and alleviated EMT in vitro and in vivo. MI remarkably inhibited tumor growth and EMT of laryngeal cancer cells in vivo.Conclusion: MI restrains the progression of laryngeal cancer and induces an inhibitory effect on EMT in laryngeal cancer by modulating the NF-κB/Twist signaling. Our finding provides new insights into the mechanism by which MI inhibits laryngeal carcinoma development, enriching the understanding of the anti-tumor function of MI.Keywords: laryngeal cancer, progression, EMT, magnesium isoglycyrrhizinate, Twist, NF-κB signaling

Vaccines induce a highly complex immune reaction in secondary lymphoid organs to generate immunological memory against an antigen or antigens of interest. Measurement of post immunization immune responses generated by specialized lymphocyte subsets requires time-dependent sampling, usually of the blood. Several T and B cell subsets are involved in the reaction, including CD4 and CD8 T cells, T follicular helper cells (Tfh), and germinal center B cells alongside their circulating (c) counterparts; cTfh and antibody secreting cells. Multicolor flow cytometry of peripheral blood mononuclear cells (PBMC) coupled with high-dimensional analysis offers an opportunity to study these cells in detail. Here we demonstrate a method by which such data can be generated and analysed using software that renders multidimensional data on a two dimensional map to identify rare vaccine-induced T and B cell subsets.

Objective The aim of this study was to evaluate the predictive factors of central lymph node metastasis (CLNM) and BRAFV600E mutation in Chinese patients with papillary thyroid carcinoma (PTC). Methods A total of 943 PTC patients who underwent thyroidectomy from 2014 to 2016 at our hospital were enrolled. Those patients were divided into PTC > 10 mm and papillary thyroid microcarcinoma (PTMC) groups by tumor size. The BRAFV600E mutation was examined by quantitative real-time PCR. Univariate and multivariate analyses were used to examine risk factors associated with CLNM and the BRAFV600E mutation. Results The frequency of CLNM was 53% (505/943). Both univariate and multivariate analyses suggested that the risk factors for CLNM in PTC patients were male, younger age, and larger tumor size (P < 0.05). Coexistent Hashimoto thyroiditis (HT) was an independent protective factor against CLNM when the tumor was > 10 mm (P = 0.006). Stratified analysis revealed that male, age ≤ 30 years, and tumor size > 5 mm were independent risk factors for CLNM. The BRAFV600E mutation rate was 85%. Multivariate logistic regression analysis revealed that age (P < 0.001) and coexistent HT (P = 0.005) were independent predictive factors of BRAFV600E mutation in PTC patients. Only age was a risk factor for the BRAFV600E mutation when the tumor was > 10 mm (P = 0.004). In the PTMC group, the BRAFV600E mutation was significantly correlated with tumor size (P < 0.001) and coexistent HT (P = 0.03). Stratified analysis revealed that age > 30 years and tumor size > 5 mm were independent predictive factors of BRAFV600E mutation. Furthermore, the incidence of CLNM was significantly higher in BRAFV600E mutation-positive patients (P = 0.009) when the tumor was ≤ 5 mm. Conclusion The factors male, younger age (≤ 30 years), large tumor size (> 5 mm), and coexistent HT are independent predicative factors for CLNM. The BRAFV600E mutation is associated with both large size and without HT in PTMC patients, age > 30 years in the PTC > 10 mm group. The BRAFV600E mutation was an independent risk factor for CLNM when the tumor was ≤ 5 mm. For optimal management, these features should be comprehensively evaluated to determine the initial surgical approach for PTC patients.

Understanding the roles played by geography and ecology in driving species diversification and in the maintenance of species cohesion is the central objective of evolutionary and ecological studies. The multi-phased orogenesis of Qinghai-Tibetan Plateau (QTP) and global climate changes over late-Miocene has profoundly influenced the environments and evolution of organisms in this region and the vast areas of Asia. In this study, we investigate the lineage diversification ofChrysanthemum-group in subtribe Artemisiinae (tribe Anthemideae, Asteraceae) likely under the effects of climate changes during this period. Using DNA sequences of seven low-copy nuclear loci and nrITS and the coalescent analytical methods, a time-calibrated phylogeny of subtribe Artemisiinae was reconstructed with emphasis onChrysanthemum-group. The monophyleticChrysanthemum-group was well resolved into two major clades corresponding toChrysanthemumandAjania, two genera which can be well identified by capitulum morphology but have been intermingled in previous plastid and ITS trees. WithinChrysanthemum, a later divergence betweenCh. indicum-complex andCh. zawadskii-complex can be recognized. The time frames of these sequential divergences coincide with the late Cenozoic uplift of the Northern QTP and the concomitant climatic heterogeneity between eastern and inland Asia. Reconstruction of historical biogeography suggested the origin ofChrysanthemum-group in Central Asia, followed by eastward migration ofChrysanthemumandin situdiversification ofAjania. WithinChrysanthemum,Ch. indicum-complex andCh. zawadskii-complex exhibited contemporary distributional division, the former in more southern and the latter in more northern regions. The geographic structure of the three lineages inChrysanthemum-group have been associated with the niche differentiation, and environmental heterogenization in Asia interior.

BackgroundPolyploid species often originate recurrently. While this is well known, there is little information on the extent to which distinct allotetraploid species formed from the same parent species differ in gene expression. The tetraploid yarrow speciesAchillea alpinaandA. wilsonianaarose independently from allopolyploidization between diploidA. acuminataandA. asiatica. The genetics and geography of these origins are clear from previous studies, providing a solid basis for comparing gene expression patterns of sibling allopolyploid species that arose independently.ResultsWe conducted comparative RNA-sequencing analyses on the twoAchilleatetraploid species and their diploid progenitors to evaluate: 1) species-specific gene expression and coexpression across the four species; 2) patterns of inheritance of parental gene expression; 3) parental contributions to gene expression in the allotetraploid species, and homeolog expression bias. DiploidA. asiaticashowed a higher contribution than diploidA. acuminatato the transcriptomes of both tetraploids and also greater homeolog bias in these transcriptomes, possibly reflecting a maternal effect. Comparing expressed genes in the two allotetraploids, we found expression of ca. 30% genes were species-specific in each, which were most enriched for GO terms pertaining to “defense response”. Despite species-specific and differentially expressed genes between the two allotetraploids, they display similar transcriptome changes in comparison to their diploid progenitors.ConclusionTwo independently originatedAchilleaallotetraploid species exhibited difference in gene expression, some of which must be related to differential adaptation during their post-speciation evolution. On the other hand, they showed similar expression profiles when compared to their progenitors. This similarity might be expected when pairs of merged diploid genomes in tetraploids are similar, as is the case in these two particular allotetraploids.

Objective:The aim of the study is to evaluate the association of the BRAFV600E mutation with the clinicopathologic characteristics in Chinese population with papillary thyroid carcinoma (PTC).MethodsA total of 943 PTC patients who underwent thyroidectomy from 2014 to 2016 at Henan Provincial People’s Hospital were included in the present study. The BRAFV600E mutation was examined in each resected specimen by quantitative Real-time PCR (qRT-PCR) technique. Results The PTC patients were subclassified into the overall, PTC>10mm and papillary thyroid microcarcinoma(PTMC) groups. The positive rate of BRAFV600E mutation was 85.4% in Chinese patients with PTC. In both overall PTC and PTC> 10mm groups, the BRAFV600E mutation was much more frequently detected in elderly patients and patients at T1 stage (P< 0.05). In addition, the positive rate of BRAFV600E mutation was significantly higher in PTC patients without concomitant Hashimoto’s thyroiditis in overall PTC and PTMC groups (P< 0.05). Furthermore, logistic regression analysis suggested that the risk of having a larger tumor diameter was increased by 6-fold when BRAFV600E mutation in the PTMC group. No association between the BRAFV600E mutation and other clinicopathologic factors was observed. ConclusionThe BRAFV600E mutation was significantly associated with patients age and T stage. Furthermore, the risk of having a larger tumor size was significantly increased when BRAFV600E mutation in the PTMC group. which suggests that BRAFV600E mutation might play an important role in the activation of early thyroid carcinogenesis, the effect might weaken in the progression of PTC.

A patient with multiple-organ echinococcosis suffered from liver echinococcosis,lung echinococcosis,and pelvic echinococcosis successively in the past three decades.From the first operation at 19 years-old,she underwent operations several times due to the recurrence of multiple organ involvement.Echinococcosis is a zoonotic disease.Although the liver usually is the primary site,the disease can also invade many other organs.Diagnosis is typically based on disease history and imaging findings.Thorough removal of the lesions during the first operation is particularly important.Comprehensive evaluations and multi-disciplinary team are helpful in the treatment of patients with multiple organ invasion.

Understanding the roles played by geography and ecology in driving species diversification and in the maintenance of species cohesion is the central objective of evolutionary and ecological studies. The multi-phased orogenesis of Qinghai-Tibetan Plateau (QTP) and global climate changes over late-Miocene has profoundly influenced the environments and evolution of organisms in this region and the vast areas of Asia. In this study, we investigate the lineage diversification of Chrysanthemum-group in subtribe Artemisiinae (tribe Anthemideae, Asteraceae) likely under the effects of climate changes during this period. Using DNA sequences of seven low-copy nuclear loci and nrITS and the coalescent analytical methods, a time-calibrated phylogeny of subtribe Artemisiinae was reconstructed with emphasis on Chrysanthemum-group. The monophyletic Chrysanthemum-group was well resolved into two major clades corresponding to Chrysanthemum and Ajania, two genera which can be well identified by capitulum morphology but have been intermingled in previous plastid and ITS trees. Within Chrysanthemum, a later divergence between Ch. indicum-complex and Ch. zawadskii-complex can be recognized. The time frames of these sequential divergences coincide with the late Cenozoic uplift of the Northern QTP and the concomitant climatic heterogeneity between eastern and inland Asia. Reconstruction of historical biogeography suggested the origin of Chrysanthemum-group in Central Asia, followed by eastward migration of Chrysanthemum and in situ diversification of Ajania. Within Chrysanthemum, Ch. indicum-complex and Ch. zawadskii-complex exhibited contemporary distributional division, the former in more southern and the latter in more northern regions. The geographic structure of the three lineages in Chrysanthemum-group have been associated with the niche differentiation, and environmental heterogenization in Asia interior.

SummaryThe ETV6-RUNX1 onco-fusion arises in utero, initiating a clinically silent pre-leukemic state associated with the development of pediatric B-acute lymphoblastic leukemia (B-ALL). We characterize the ETV6-RUNX1 regulome by integrating chromatin immunoprecipitation- and RNA-sequencing and show that ETV6-RUNX1 functions primarily through competition for RUNX1 binding sites and transcriptional repression. In pre-leukemia, this results in ETV6-RUNX1 antagonization of cell cycle regulation by RUNX1 as evidenced by mass cytometry analysis of B-lineage cells derived from ETV6-RUNX1 knock-in human pluripotent stem cells. In frank leukemia, knockdown of RUNX1 or its co-factor CBFβ results in cell death suggesting sustained requirement for RUNX1 activity which is recapitulated by chemical perturbation using an allosteric CBFβ-inhibitor. Strikingly, we show that RUNX1 addiction extends to other genetic subtypes of pediatric B-ALL and also adult disease. Importantly, inhibition of RUNX1 activity spares normal hematopoiesis. Our results implicate chemical intervention in the RUNX1 program as an exciting therapeutic opportunity in ALL.

Evolutionary shifts among radiate, disciform and discoid flowerheads have occurred repeatedly in a number of major lineages across the Asteraceae phylogeny; such transitions may also appear within evolutionarily young groups. Although several studies have demonstrated that CYC2 genes partake in regulating floral morphogenesis in Asteraceae, the evolution of capitulum forms within a recently diverging lineage has remained poorly understood. Here, we study the molecular regulation of the shift from a radiate to a disciform capitulum within the Chrysanthemum group. This is a recently radiating group mainly comprising two genera, Chrysanthemum and Ajania, that are phylogenetically intermingled but distinct in flowerhead morphology: Chrysanthemum spp. with radiate capitula and Ajania spp. with disciform capitula. We found that the morphogenesis of zygomorphy in the marginal floret in Ajania was disrupted soon after floral primordium emergence; CYC2g, one of the CYC2 copies that was expressed prominently in the ray floret of Chrysanthemum was not expressed in flowerheads of Ajania. Weakening the expression of ClCYC2g in Chrysanthemum lavandulifolium led to the gradual transition of a ray flower toward the disc-like form. Molecular evolutionary analyses indicated that the disciform capitulum might have evolved only once, approximately 8 Mya, arising from dysfunction of the CYC2g orthologs. A 20-nt deletion, including a putative TATA-box of the Ajania-type CYC2g promoter, appeared to inhibit the expression of the gene. Considering the divergent habitats of Chrysanthemum and Ajania, we propose that the shift from radiate to disciform capitulum must have been related to changes in pollination strategies under selective pressure.

The immunoglobulin capture assay (ICA) enables the enrichment for pathogen-specific plasmablasts from individuals with a confirmed adaptive immune response to vaccination or disseminated infection. Only single recombinant antigens have been used previously as probes in this ICA and it was unclear whether the method was applicable to complex probes such as whole bacterial cells. Here, we describe the enrichment of plasmablasts specific for polysaccharide and protein antigens of both Streptococcus pneumoniae and Neisseria meningitidis using whole formalin-fixed bacterial cells as probes. The modified ICA protocol described here allowed for a pathogen-specific hmAb cloning efficiency of >80%.

Recently, the layered transition metal dichalcogenide 1T′-MoTe2 has generated considerable interest due to their superconducting and non-trivial topological properties. Here, we present a systematic study on 1T′-MoTe2 single-crystal and exfoliated thin-flakes by means of electrical transport, scanning tunnelling microscope (STM) measurements and band structure calculations. For a bulk sample, it exhibits large magneto-resistance (MR) and Shubnikov–de Hass oscillations in ρxx and a series of Hall plateaus in ρxy at low temperatures. Meanwhile, the MoTe2 thin films were intensively investigated with thickness dependence. For samples, without encapsulation, an apparent transition from the intrinsic metallic to insulating state is observed by reducing thickness. In such thin films, we also observed a suppression of the MR and weak anti-localization (WAL) effects. We attributed these effects to disorders originated from the extrinsic surface chemical reaction, which is consistent with the density functional theory (DFT) calculations and in-situ STM results. In contrast to samples without encapsulated protection, we discovered an interesting superconducting transition for those samples with hexagonal Boron Nitride (h-BN) film protection. Our results indicate that the metallic or superconducting behavior is its intrinsic state, and the insulating behavior is likely caused by surface oxidation in few layer 1T’-MoTe2 flakes.

Mg–Y–Zn alloys with different contents of alloying elements were prepared. The microstructure and mechanical properties of these Mg–Y–Zn alloys were investigated by using scanning electron microscopy, X-ray diffraction and tensile testing. The results show that the as-cast Mg–Y–Zn alloy is mainly composed of α-Mg, Mg12YZn and Mg3Y2Zn3 phases. After Mn addition, a small quantity of Mn phase arises in the as-cast Mg–Y–Zn–Mn alloy. After extrusion, the microstructure of the alloys is of an inhomogeneous character showing fine grains produced due to dynamic recrystallization. Tensile strength of as-extruded Mg-9Y-1Zn, Mg-9Y-3Zn and Mg-9Y-3Zn-1Mn reached 249.86 MPa, 303.19 MPa and 322.15 MPa, respectively. The gradually increasing strength can be attributed to fine grain strengthening, second phase strengthening and LPSO structure. Additionally, the tensile fracture surfaces were examined under scanning electron microscopy, and all of their fracture morphologies exhibit dimple facture character.

Many studies have addressed the desorption and mobilization performances of sorbed contaminants affected by different rhamnolipidic biosurfactants. Study results have been mixed and complicated. Rhamnolipids are always microbial produced with variable homologues. In this study, two representative rhamnolipidic fractions (i.e., RL-F1 and RL-F2, which are mono- and di-rhamnolipids, respectively) were investigated and compared to determine their influence on 17α-ethynylestradiol (EE2) distribution within sediment-water sorption and desorption systems. In general, the coexistence of RL-F1 and EE2 enhanced EE2 sorption in a wider monorhamnolipidic dosage range when freshly treated sorbate was used. The sorbed EE2 concentration decreased as the RL-F1 dosage increased in the aged sorbate desorption systems. However, RL-F2 facilitated EE2 mobilization in both sorption and desorption processes. Experimental data were estimated using a conceptual model that considered the sorbed rhamnolipids and aqueous micelles for organic partitioning. The model results indicated that the rhamnolipid type is an important factor influencing organic distribution, in addition to sorbate aging process and sediment characteristics. The use of a rhamnolipidic mixture containing both mono- and di-rhamnosyl components may not achieve the desired effect when the biosurfactant-enhanced mobilization or immobilization approach is selected. These results are significant for selecting and applying rhamnolipids to remediate contaminants.

Background and aims To understand the link between species diversity and phenotype developmental evolution is an important issue in evolutionary biology. Yarrows in the genus Achillea (Asteraceae) show a great diversity in leaf serrate or pinnate dissection patterns. In Arabidopsis thaliana, the development of leaf serration requires the activity of the transcription factor CUC2. Does this regulator also work for leaf dissections of the Asteraceae plants? If so, how do the conserved regulatory 'tools' work differently to produce diverse leaf forms? Methods Seedling leaf morphology was observed, and morphogenesis of leaf serration or lobes was examined by scanning electron microscopy (SEM). NAM genes, orthologues of arabidopsis CUC2, were isolated from A. acuminata with serrate leaves and A. asiatica with three-pinnatisect leaves, respectively. By means of whole-mount in situ mRNA hybridization and two quantitative gene expression assays, the droplet digital PCR (ddPCR) and quantitative real-time PCR (qPCR), expression patterns of the NAM genes during leaf dissection development were checked in both species for comparison. Key results For both species, the development of leaf dissection initiated when a leaf blade was about 300-400 µm long. In A. acuminata, in situ hybridization showed NAM expression signals at leaf margins where teeth are growing, or later on, in the sinuses of the teeth, whilst in A. asiatica, hybridization signals appear not only on leaf margins but further on the margins of leaf lobes. Both ddPCR and qPCR revealed a continuous decline of AacNAM expression from the early to the late developmental stages of a single leaf of A. acuminata, whereas a relatively long maintenance and fluctuation of AasNAM expression was seen in a leaf of A. asiatica. Conclusions Differential spatiotemporal patterns of NAM expression were found between the two yarrow species during development of leaf dissection. This study provides the first evidence for NAM activity in the development of leaf dissection of the Asteraceae plants, and demonstrates that leaf form diversity is correlated to the altered NAM expression dynamic.

We report the development of a novel flow cytometry–based Ig capture assay (ICA) for the identification and sorting of individual Ab-secreting cells based on their Ag reactivity. The ICA represents a fast and versatile tool for single-cell sorting of peripheral plasmablasts, streamlining subsequent Ab analysis, and cloning. We demonstrate the utility of the assay by isolating Ag-reactive plasmablasts from cryopreserved PBMC obtained from volunteers vaccinated with a recombinant HIV envelope protein. To show the specificity of the ICA, we produced Ag-specific Abs from these cells and subsequently verified their Ag reactivity via ELISA. Furthermore, we used the ICA to track Ag-specific plasmablast responses in HIV-vaccine recipients over a period of 42 d and performed a head-to-head comparison with a conventional B cell ELISpot. Results were highly comparable, highlighting that this assay is a viable alternative for monitoring Ag-specific plasmablast responses at early time points after infection or vaccination. The ICA provides important added benefits in that phenotypic information can be obtained from the identified Ag-specific cells that can then be captured for downstream applications such as B cell sequencing and/or Ab cloning. We envisage the ICA as being a useful tool in Ab repertoire analysis for future clinical trials.

BACKGROUND Therapeutic patient education is a continuous, systematic, patient-centered learning process to help patients and their families acquire and maintain the skills they need to manage their lives with a chronic disease. It has been proven effective in increasing treatment adherence and improving quality of life for patients with atopic dermatitis (AD) in Western countries. We introduce the first multicenter, randomized controlled clinical trial of therapeutic patient education in Chinese children with AD. OBJECTIVES To evaluate the effects of therapeutic patient education on the severity of AD, quality of life, and understanding and successful use of emollients in Chinese children with eczema. METHODS We recruited 580 children, ages 2-14 years, with moderate to severe AD from six hospitals in China. Participants were randomized (1:1) to an intervention (n = 293) or control (n = 249) group. In addition to the severity of AD, data on quality of life and a questionnaire on family and patient knowledge of emollients were evaluated at the 6-month follow-up. RESULTS On study completion, we found that the intervention group showed a significantly greater reduction in mean SCORing Atopic Dermatitis (P < .001) and Infant's Dermatology Life Quality Index (P = .030) scores than the control group. In addition, knowledge about the use of emollients improved significantly in the intervention group. There was no significant difference between groups in Children's Dermatology Life Quality Index scores. CONCLUSIONS The first randomized controlled trial of a therapeutic patient education program in China had positive long-term effects on decreasing eczema severity and improvement of quality of life in children 2-4 years of age with AD, as well as in promoting greater understanding of the use of emollients.

Objective To investigate whether topical mucopolysaccharide polysulfate (MPS) cream can reduce the incidence of eczema and skin atrophy in patients with moderate- or low-risk infantile hemangioma after the treatment with topical beta-blockers or 595-nm pulsed dye laser (PDL) , and to analyze factors influencing the occurrence of eczema and skin atrophy. Methods A total of 722 patients aged 0- 1 years with moderate- or low-risk infantile hemangioma were enrolled from 5 Children′s Hospitals in China. According to the disease condition and therapy acceptability, these patients were divided into 6 groups to be treated with topical beta-blockers and MPS cream (blocker+ MPS group) , topical beta-blockers (blocker group) , 595-nm PDL and topical MPS cream (PDL+ MPS group) , 595-nm PDL (PDL group) , 595-nm PDL combined with topical beta-blockers and MPS cream (PDL+ blocker+ MPS group) , and 595-nm PDL and topical beta-blockers (PDL+ blocker group) , respectively. All the externally applied agents were applied twice a day, and PDL was performed once every 4 weeks. Efficacy and adverse reactions were evaluated after 3-month treatment. Univariate and multivariate Logistic regression analyses were carried out to analyze factors influencing the incidence of eczema and skin atrophy in patients with infantile hemangioma after treatment, and chi-square test was carried out to compare efficacy among the groups. Results After 3-month treatment, multivariate Logistic regression analysis for comparing the blocker+ MPS group with blocker group showed that the risk factor for eczema on the surface of hemangiomas was no topical treatment with MPS cream (P= 0.007, OR= 3.887, 95% CI: 1.439-10.493) , while no correlations were observed between the occurrence of skin atrophy on the surface of hemangiomas and analyzed factors. Multivariate Logistic regression analysis for comparing the PDL+MPS group with PDL group showed that no topical treatment with MPS cream (P 0.05) . Conclusion During the treatment of moderate- or low-risk infantile hemangioma with topical beta-blockers or 595-nm PDL, the combination with topical MPS cream can reduce the occurrence of eczema and skin atrophy without affecting the therapeutic effect. Key words: Hemangioma; Infant; Eczema; Mucopolysaccharide polysulfate; Skin atrophy; Adverse reaction

T cell help for B cells may be perturbed in people living with HIV (PLWH), even when HIV is suppressed, as evidenced by reports of suboptimal responses to influenza vaccination. We investigated cTFH responses to the 2017–18 inactivated quadrivalent influenza vaccine (QIV) in men living with antiretroviral therapy (ART)-suppressed HIV infection who were treated in the early or chronic phase of infection, and control subjects. Here we show that seroprotective antibody responses in serum and oral fluid correlated with cTFH activation and were equivalent in all three groups, irrespective of when ART was started. These responses were attenuated in those reporting immunisation with influenza vaccine in the preceding three years, independent of HIV infection. Measurement of influenza-specific IgG in oral fluid was closely correlated with haemagglutination inhibition titre. T-SNE and two-dimensional analysis revealed a subset of CD4+CXCR3+CXCR5+ cTFH activated at one week after vaccination. This was distinguishable from cTFH not activated by vaccination, and a rare, effector memory CD4+CXCR5hiCD32hi T cell subset. The data support the use of QIV for immunisation of PLWH, reveal distinct circulating CD4+CXCR5+ T cell subsets and demonstrate oral fluid sampling for influenza-specific IgG is an alternative to phlebotomy.

This 14‐color, 16‐antibody OMIP was designed for enumeration of leukocyte responses in pediatric samples, where sample volumes and cell numbers can be very low. Leukocytes identified by this panel include all major members of the innate lymphoid cell (ILC) family (ILC1s, ILC2s, and ILC3s), natural killer cells (NK cells), granulocytes (neutrophils and eosinophils), T‐cells (CD4+ and CD8+), mucosal‐associated invariant T cells (MAIT cells) and NKT‐like cells. The protocol was optimized using small volumes of peripheral blood and validated in airway samples obtained from children (

Background Autosomal dominant optic atrophy (ADOA) is usually caused by mutations in the essential gene, OPA1. This encodes a ubiquitous protein involved in mitochondrial dynamics, hence tissue specificity is not understood. Dysregulated mitophagy (mitochondria recycling) is implicated in ADOA, being increased in OPA1 patient fibroblasts. Furthermore, autophagy may be increased in retinal ganglion cells (RGCs) of the OPA1Q285STOP mouse model. Aims We developed a mouse model for studying mitochondrial dynamics in order to investigate mitophagy in ADOA. Methods We crossed the OPA1Q285STOP mouse with our RedMIT/GFP-LC3 mouse, harboring red fluorescent mitochondria and green fluorescent autophagosomes. Colocalization between mitochondria and autophagosomes, the hallmark of mitophagy, was quantified in fluorescently labeled organelles in primary cell cultures, using two high throughput imaging methods Imagestream (Amnis) and IN Cell Analyzer 1000 (GE Healthcare Life Sciences). We studied colocalization between mitochondria and autophagosomes in fixed sections using confocal microscopy. Results We validated our imaging methods for RedMIT/GFP-LC3 mouse cells, showing that colocalization of red fluorescent mitochondria and green fluorescent autophagosomes is a useful indicator of mitophagy. We showed that colocalization increases when lysosomal processing is impaired. Further, colocalization of mitochondrial fragments and autophagosomes is increased in cultures from the OPA1Q285STOP/RedMIT/GFP-LC3 mice compared to RedMIT/GFP-LC3 control mouse cells that were wild type for OPA1. This was apparent in both mouse embryonic Diot et al. Mouse Model for Detecting Mitophagy fibroblasts (MEFs) using IN Cell 1000 and in splenocytes using ImageStream imaging flow cytometer (Amnis). We confirmed that this represents increased mitophagic flux using lysosomal inhibitors.We also used microscopy to investigate the level of mitophagy in the retina from the OPA1Q285STOP/RedMIT/GFP-LC3 mice and the RedMIT/GFP-LC3 controlmice. However, the expression levels of fluorescent proteins and the image signalto- background ratios precluded the detection of colocalization so we were unable to show any difference in colocalization between these mice. Conclusions We show that colocalization of fluorescent mitochondria and autophagosomes in cell cultures, but not fixed tissues from the RedMIT/GFP-LC3, can be used to detect mitophagy. We used this model to confirm that mitophagy is increased in a mouse model of ADOA. It will be useful for cell based studies of diseases caused by impaired mitochondrial dynamics.

In our study, the effects of water stress on photosynthesis and photosynthetic electron transport chain (PETC) were studied in several ways, including monitoring the change of gas exchange parameters, modulated chlorophyll fluorescence, rapid fluorescence induction kinetics, reactive oxygen species (ROS), antioxidant enzyme activities and D1 protein levels in apple leaves. Our results show that when leaf water potential (ψw) is above –1.5 MPa, the stomatal limitation should be the main reason for a drop of photosynthesis. In this period, photosynthetic rate (PN), stomatal conductance (Gs), transpiration rate (E) and intercellular CO2 concentration (Ci) all showed a strong positive correlation with ψw. Modulated chlorophyll fluorescence parameters related to photosynthetic biochemistry activity including maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency of PSII (ΦPSII), photochemical quenching coefficient (qP) and coefficient of photochemical fluorescence quenching assuming interconnected PSII antennae (qL) also showed a strong positive correlation as ψw gradually decreased. On the other hand, in this period, Stern-Volmer type non-photochemical quenching coefficient (NPQ) and quantum yield of light-induced non-photochemical fluorescence quenching [Y(NPQ)] kept going up, which shows an attempt to dissipate excess energy to avoid damage to plants. When ψw was below –1.5 MPa, PN continued to decrease linearly, while Ci increased and a ‘V’ model presents the correlation between Ci and ψw by polynomial regression. This implies that, in this period, the drop in photosynthesis activity might be caused by non-stomatal limitation. Fv/Fm, ΦPSII, qP and qL in apple leaves treated with water stress were much lower than in control, while NPQ and Y(NPQ) started to go down. This demonstrates that excess energy might exceed the tolerance ability of apple leaves. Consistent with changes of these parameters, excess energy led to an increase in the production of ROS including H2O2 and O2•−. Although the activities of antioxidant enzymes like catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD) increased dramatically and ascorbate peroxidase (APX) decreased in apple leaves with drought stress, it was still not sufficient to scavenge ROS. Consequently, the accumulation of ROS triggered a reduction of net D1 protein content, a core protein in the PSII reaction center. As D1 is responsible for the photosynthetic electron transport from plastoquinone A (QA) to plastoquinone B (QB), the capacity of PETC between QA and QB was considerably downregulated. The decline of photosynthesis and activity of PETC may result in the shortage of adenosine triphosphate (ATP) and limitation the regeneration of RuBP (Jmax), a key enzyme in CO2 assimilation. These are all non-stomatal factors and together contributed to decreased CO2 assimilation under severe water stress., Summary: This research reports that drought-induced photosynthesis declines due to stomatal and non-stomatal limitation, which was PSII photoinhibition-dependent on D1 protein and over-reduces the electron transport chain.

Cervical spondylosis (CS), which is resulted from degeneration of cervical intervertebral disc, is a common disease seriously threatening human health and quality of life. However, there is still no effective clinic strategies for the treatment of this disease. The acupoint stimulation with needle-scalpel is a widely used approach to treat orthopedic diseases. In the present study, we evaluated the therapeutic effects of acupoint stimulation around neck with needle-scalpel on delaying the degeneration of cervical intervertebral discs and hopefully provided an approach for the precaution and early intervention of CS. We firstly established a rat model of CS by cervical static-dynamic imbalance to mimics disc degeneration and then stimulated the acupoints around neck with needle-scalpel. The cervical intervertebral disc samples were collected to measure type I and II collagen by quantitative PCR (qPCR), immunohistochemistry, and western blot. The changes in micro-structure and ultra-structure of nucleus pulposus were analyzed under the optical microscope and electron microscope respectively. Acupoint stimulation with needle-scapelon increased type I collagen production and decreased type II collagen production, and improved the micro-structure and ultra-structure of nucleus pulposus. Our results suggest that acupoint stimulation around neck with needle-scapelon could inhibit intervertebral disc degeneration through modulating the extracellular matrix collagen system and improving the changed structure of nucleus pulposus.

This article reports a female type Ⅱ Lynch syndrome patient who suffered from colon cancer and endometrium carcinoma consecutively. We also briefly elucidate the pathogenesis,clinicopathological features,screening,and management of Lynch syndrome. Regular screening and early diagnosis and treatment may help to reduce the incidences and mortality rates of tumors associated with Lynch syndrome.

There are several types of capitulum in the Asteraceae due to different combinations of florets varying in corolla shape and stamen development. Previous studies have shown that the formation of ray florets on a radiate capitulum may be related to the parallel co-option of

Summary We explore cell heterogeneity during spontaneous and transcription-factor-driven commitment for network inference in hematopoiesis. Since individual genes display discrete OFF states or a distribution of ON levels, we compute and combine pairwise gene associations from binary and continuous components of gene expression in single cells. Ddit3 emerges as a regulatory node with positive linkage to erythroid regulators and negative association with myeloid determinants. Ddit3 loss impairs erythroid colony output from multipotent cells, while forcing Ddit3 in granulo-monocytic progenitors (GMPs) enhances self-renewal and impedes differentiation. Network analysis of Ddit3-transduced GMPs reveals uncoupling of myeloid networks and strengthening of erythroid linkages. RNA sequencing suggests that Ddit3 acts through development or stabilization of a precursor upstream of GMPs with inherent Meg-E potential. The enrichment of Gata2 target genes in Ddit3-dependent transcriptional responses suggests that Ddit3 functions in an erythroid transcriptional network nucleated by Gata2., Graphical Abstract, Highlights • We present a method for inferring gene regulatory networks (GRNs) from single cells • Lineage cross-antagonism is a key property of GRNs of early lineage commitment • Ddit3 is a regulatory node in erythroid lineage programming • A Ddit3-Gata2 regulatory axis antagonizes myeloid and enables erythroid programs, Pina et al. develop a gene regulatory network inference method using single-cell gene expression data and identify Ddit3 as a regulatory node in erythroid lineage programming. The authors explore this inference and show that Ddit3 can antagonize myeloid programming and enable erythroid signatures and forms a regulatory axis with Gata2.

Asteraceae are distinguishable by their head-like inflorescence called a capitulum. There are a variety of capitulum types due to different combinations of different composing florets. Morphological evolution of capitula must involve homoplasy under selective constraints. Therefore, a comparative examination of ontogenies of divergent capitula among phylogenetically close taxa is important to investigate the genetic and molecular bases of this trait. In the subtribe Artemisiinae, tribe Anthemideae, three closely related genera have different flower heads: Chrysanthemum with the radiate, Ajania with the disciform, and Stilpnolepis with the discoid capitula. Through observations using scanning electron microscopy, we examined the morphogenetic processes of flower heads of representative species of these genera. The morphological differentiation of the discoid from the radiate/disciform capitula occurred early in the floral initiation stage. The development of the ray/marginal florets of the radiate/disciform capitula lagged behind the outermost disc florets throughout the phases of floral initiation and floral organogenesis. Primordia of their 1–2 dorsal corolla lobes and all stamens ceased to grow soon after initiation. In contrast, all florets on a discoid capitulum developed equally in rates and modes into bisexual actinomorphic flowers. Differentiation between the radiate and the disciform capitula occurred late during the floral maturation stage. Our data provided evidence to the interpretation that uniformly acropetal development only occurs in species with homogamous flower heads, and to the idea that the timing of ontogenetic divergence between taxa is positively correlated with the phylogenetic distance of the taxa.

SnSe has emerged as an efficient thermoelectric material since a high value of the thermoelectric figure of merit ($ZT$) has been reported recently. Here we show with systematic angle resolved photoemission spectroscopy data that the low-lying electronic structures of undoped and hole-doped SnSe crystals exhibit noticeable temperature variation from 80 to 600 K. In particular, the hole effective masses for the two lowest lying valence band maxima are found to be very large and increase with decreasing temperature. Thermoelectric parameters derived from such hole-mass enhancement agree well with the transport values, indicating comprehensively a reduced impact of multivalley transport to the system's thermoelectric performance.

A novel topology for efficient utilization of parallel inverters as current source active power filter (APF) for high-power applications is presented and analyzed. The proposed technique operates the master inverter with high-power low-switching-frequency devices to compensate the low-order large-amplitude current harmonic components and the slave inverter with low-power high-switching-frequency devices to compensate the high-order small-amplitude current harmonic components. This paper discusses the operating principle, main circuit and control system design. Simulation and experimental results are provided to demonstrate the viability of the scheme.

In order to control the plasticity of CaO-SiO2-Al2O3system inclusions so as to reduce the harm to the quality of 42CrMoA crankshaft steel, the target controlling ranges of CaO-SiO2-Al2O3inclusions is put forward. On the basis, influence of CaO-SiO2-Al2O3and CaO-SiO2-Al2O3-MgO system top slag on the content of calcium and aluminum in molten steel is contrasted. Theoretical research results show that, calcium activity should be controlled in the range of (90~3300)×10-10, oxygen activity should be below 7×10-6, the mass percent of calcium should be controlled more than 75ppm. And CaO-SiO2-Al2O3-MgO should be a good choice to obtain the plastic inclusions of CaO-SiO2-Al2O3.

This paper presents a novel single-point rolling bearing fault diagnosis mechanism through vibration signal analysis. It is highlighted that the rolling bearing operational state can be well estimated by the first small set of Intrinsic Mode Function (IMF) components of the original vibration measurements through Empirical Mode Decomposition (EMD). These IMF components can be further translated into envelope spectrum by using Hilbert Transform. As a result, the difference of fault characteristic frequencies (DFCF) is derived to properly characterize different fault patterns for fault diagnosis. The suggested method is implemented and evaluated in a rolling bearing test bed for a range of failure scenarios (e.g. inner and outer raceway fault, rolling elements fault) with extensive vibration measurements. The result demonstrates that the proposed solution is effective for characterizing and detecting arrange of rolling bearing faults.quality).

Leukemic lymphoblasts within different immunophenotypic populations possess stem cell properties. However, whether or not the self-renewal program is retained from stem cells or conferred on progenitors by leukemogenic molecules remains unknown. We have addressed the issue in the context of TEL-AML1-associated acute lymphoblastic leukemia (ALL) by profiling a refined program edited from genes essential for self-renewal of hematopoietic stem cells and B-cell development. Bioinformatic analysis shows that ALL populations are loosely clustered and close to the normal population that contains stem and primitive progenitor cells. This finding indicates that immunophenotypes do not reflect maturation stages in ALL and that the self-renewal program may be retained from stem cells. Results of assessing 'first hit' function of TEL-AML1 in different populations of normal cells demonstrate the molecular model. Therefore, the current study shows a leukemogenic scenario of human ALL in which programs of stem cells are sustained in distinct fractions by leukemogenic mutations.

The glass-ceramics of CaO-Al2O3-SiO2system were prepared by sintering using lead-zinc tailings. Orthogonal experiment was carried out to optimize the ratio of lead-zinc tailings, chemical composition of basic-glass and other chemical materials in the system. Crystallization heat treatment conditions were confirmed by the differential thermal analysis (DTA) curve of glass aggregate. Crystalline phase, crystal morphology and mechanical property of the glass-ceramics samples were studied by means of X-ray diffraction (XRD), scanning electron microscope (SEM), hardness and bending strength test. The results indicated that the optimized ratio of the samples was lead-zinc tailings 25%, quartz 46.63%, calcite 21.05%, Al2O310.19% and nucleating agent TiO21%. The optimized crystallization heat treatment was that basic glass was first heated to 800 °C at 10 °C/min, then 800°C-900 °C at 5 °C/min and held still at 900 °C for 2 h. After that, the basic glass was heated to 1230 °C at 1-1.5 °C/min,held for 1h. At last, the glass-ceramics was obtained after the cooling-down of the system. The main crystalline phase of microcrystal glass-ceramics are diopside. The hardness and bending strength were 9.14 GPa and 213.6 MPa, respectively. The glass-ceramics had excellent chemical durability in acid and alkali solution, with weight losses of 0.17% and 0.68%, respectively.

The shake flask aerobic biodegradation experiment was used to study the effect on biodegradation of 17α-ethinylestradiol (EE2) in the sediment/water system affected by mono-rhamnolipid (RL-F1) and di-rhamnolipid (RL-F2). The results showed that RL-F1 and RL-F2 had no inhibitory effect on EE2 biodegradation. The effect of RL-F1 on enhanced biodegradation of EE2 was larger than that of RL-F2. In 6 mM and 10 mM of the rhamnolipidic functioned systems, EE2 biotransformation rates affected by RL-F1 were respectively 7.6 and 8.2 times of those of RL-F2 corresponding systems. The accumulation and change rule of the intermediate metabolites M.1 with the polarity greater than EE2 and M.3 with the polarity weaker then EE2 in the biodegradation process were analyzed. In the RL-F2 systems, the time for the accumulative amounts of M.1 and M.3 up to the peak was correspondingly lagged and their output was relatively lower. The testing result of rhamnolipids in the sample showed that RL-F1 was more biodegradable than RL-F2. The bioavailability of di-rhamnolipidic micelle solubilized with EE2 was inferior to that of mono-rhamnolipid, making the organic uptake and metabolism process of microorganisms slow down.

Rhamnolipids are typically produced as multicomponent mixtures of homologues and are notably promising for environmental remediation. Because different mixtures exhibit dissimilar surface-active and micellization properties, which may affect their functional behaviors, the individual contribution of each rhamnolipidic homologue to the solubilization has largely been unexplored. In this study, enhanced solubilization of moderately hydrophobic 17α-ethinylestradiol (EE2) by two independent rhamnolipidic fractions (i.e., RL-F1 and RL-F2, which are mono- and di-rhamnolipids, respectively) was investigated. The results of the solubilization experiments indicated that RL-F2 exhibited a higher maximum dissolving capability of EE2 compared to RL-F1. However, a higher mass transfer rate was observed for RL-F1. To confirm the results of the solubilization experiments, dynamic light scattering, transmission electron microscopy, and UV-spectral analysis were performed. The type of rhamnolipid and the aggregate morphology strongly influenced the incorporation of solubilizate into the surfactant aggregates. The results of this study may be used to understand the rhamnolipid-facilitated transport of contaminants in aquatic environments, as well as the selection of rhamnolipids, with high specificity for practical applications.

The aim of the present work was to develop a lain-human finite element model for cushion design to prevent bedsore by performing biomechanical analysis on interface pressure. The geometric data of the human was obtained by laser scans. The finite element model was composed of solid hexahedral elements. The material of the bed cushions was obtained according to the mechanical experiment. The human model was validated by comparing the simulation result with the experimental data. The validated finite element model could be used to facilitate, accelerate and economize the process of design of cushion.

The zinc finger transcription factor GATA-2 has been implicated in the regulation of hematopoietic stem cells. Herein, we explored the role of GATA-2 as a candidate regulator of the hematopoietic progenitor cell compartment. We showed that bone marrow from GATA-2 heterozygote (GATA-2+/−) mice displayed attenuated granulocyte-macrophage progenitor function in colony-forming cell (CFC) and serial replating CFC assays. This defect was mapped to the Lin−CD117+Sca-1−CD34+CD16/32high granulocyte-macrophage progenitor (GMP) compartment of GATA-2+/− marrow, which was reduced in size and functionally impaired in CFC assays and competitive transplantation. Similar functional impairments were obtained using a RNA interference approach to stably knockdown GATA-2 in wild-type GMP. Although apoptosis and cell-cycle distribution remained unperturbed in GATA-2+/− GMP, quiescent cells from GATA-2+/− GMP exhibited altered functionality. Gene expression analysis showed attenuated expression of HES-1 mRNA in GATA-2–deficient GMP. Binding of GATA-2 to the HES-1 locus was detected in the myeloid progenitor cell line 32Dcl3, and enforced expression of HES-1 expression in GATA-2+/− GMP rectified the functional defect, suggesting that GATA-2 regulates myeloid progenitor function through HES-1. These data collectively point to GATA-2 as a novel, pivotal determinant of GMP cell fate.

The geometrically frustrated spin-1 Blume-Capel model on an infinite sawtooth chain is exactly solved by the transfer matrix method. The magnetization, ground-state phase diagram, magnetocaloric properties, and specific heat of the system are investigated. The results indicate that: (i) Magnetization plateaus appear at zero temperature. Their number depends on the sign of the crystal field D. For D≥0 there are two magnetization plateaus; however, for D

Standard programmable logic controller (PLC) based proportional-integral-derivative (PID) was originally employed as part of the food production line control strategy, but after observing the response of those process due to measured disturbances during normal operation, it was evident that PID control could not meet the desired cooking specifications. It was decided to pursue advanced process control strategies as a means to meet the food production specifications. The problem of this research study was to design and analyze the performance of a PLC-based model state feedback controller implementation for an industrial food production line, and to determine its viability in comparison to commercially available PC-based model predictive controller implementations applied to food production line.

To observe the effect of acupotomy lysis at acupoints around the neck on expression of matrix metalloproteinase 1 (MMP-1), MMP-2 and tissue inhibitor of metalloproteinase 1 (TIMP-1) genes and ultrastructure of pulpiform nucleus in cervical intervertebral disc degeneration (IVDD) rats, so as to explore its mechanism underlying easing IVDD.SD rats were randomly allocated to control (n = 15), model (n = 14), Jiaji (EX-B 2, n = 13), cervico-acupoint (n = 14) and medication groups (n = 14). The cervical IVDD model was established by using static-dynamic imbalance method. For rats of the Jiaji (EX-B 2) and cervico-acupoint groups, EX-B 2-points of the cervical 2-7 segments, and peri-cervical acupoints: bilateral "Naokong" (GB 19) , "Naohu" (GV 17), "Dazhui" (GV 14), bilateral "Quyuan" (SI 13) and bilateral "Tianzong" (SI 11) were separately punctured with a needle-knife, once every 5 days for 3 times, and for rats of the medication group, Brufen Capsules (15 mg · kg(-1) · d(-1)) and Jingfukang Granule (0.5 mg · kg(-1) · d(-1)) were given by intragastric administration, once daily for 10 days. The expression levels of MMP-1, MMP-3 and TIMP-1 genes in the pulpiform nucleus of cervical intervertebral discs were detected by RT-PCR and changes of the ultrastructure of the pulpiform nucleus observed under transmission electron microscope.Compared to the control group, the expression levels of MMP-1 mRNA and MMP-3 mRNA of the cervical intervertebral disc tissues were significantly up-regulated in the model group (P0.05), and that of TIMP-1 mRNA was obviously down-regulated in the model group (P0.05). After the treatment, the increased expression of MMP-1 mRNA and MMP-3 mRNA and the decreased expression of TIMP-1 mRNA were reversed by acupotomy lysis and medication (P0.05) except TIMP-1 mRNA in the medication group (P0.05). No significant differences were found between the Jiaji (EX-B 2) and cervico-acupoint groups in down-regulating MMP-1 mRNA and MMP-3 mRNA expression and up-regulating TIMP-1 mRNA expression (P0.05). Results of electron microscope examinations showed that the ultrastructural injury changes of cells of the pulpiform nucleus were relatively milder in the Jiaji (EX-B 2) and cervico-acupoint groups, followed by the medication group in comparison with those of the model group.Acupotomy lysis at acupoints around the neck can improve the ultrastructural changes of cells of the pulpiform nucleus of cervical intervertebral discs in IVDD rats, which is possibly by regulating the expression of MMP-1, MMP-3 and TIMP-1 genes.

The genus Leontopodium comprises 30-41 species. The centre of diversity is the Sino-Himalayan region in south-western China, where about 15 species occur. The two species native to Europe, L. alpinum (known as the common 'Edelweiss') and L. nivale, are part of the cultural heritage of the people living there. Despite its importance, very little is known about the systematics of the genus. Because recent molecular studies have shown that species within this genus are closely related and difficult to distinguish with rDNA and cpDNA data, we used AFLPs to obtain a more detailed understanding of the phylogeny of the genus. Our main aims were as follows: (1) to clarify species relationships within the genus; and (2) to reveal information about the biogeography of the genus. We used AFLPs with six primer combinations to investigate 216 individuals in 38 populations of 16 different species. With AFLPs, we were able to recognize 10 different groups, all of which had strong bootstrap support. These results were also congruent with the morphology-based taxonomy of the genus. Most private and rare fragments were found in the Yunnan region (south-western China) relative to Europe and Mongolia/central China, suggesting a long-lasting in situ history of populations in the centre of diversity of the genus. Our results illustrate the utility of AFLPs to resolve phylogenetic relationships between these closely related species.