Effects of Toutongning capsule on enkephalin expression in a rat migraine headache model

Toutongning capsule is used for the treatment of migraine headaches, and has yielded therapeutically beneficial effects. However, whether Toutongning capsule exerts its effects via endogenous opioid peptides remains poorly understood. This study investigated the effects of Toutongning capsule on enkephalin expression in the midbrain of rats with nitroglycerin-induced migraine headache at the mRNA and protein levels. Results confirmed that proenkephalin mRNA levels were significantly upregulated following treatment with Toutongning capsule. The numbers methionine-enkephalin and leucine-enkephalin-positive cells were significantly greater in the migraine headache rats following treatment with Toutongning capsule compared with the model group. Taken together, these results show that Toutongning capsule exerts potentially beneficial effects by promoting enkephalin expression in nitroglycerin-induced migraine headache rats.

CitGVD:a comprehensive database of citrus genomic variations

Citrus is one of the most important commercial fruit crops worldwide.With the vast genomic data currently available for citrus fruit,genetic relationships,and molecular markers can be assessed for the development of molecular breeding and genomic selection strategies.In this study,to permit the ease of access to these data,a web-based database,the citrus genomic variation database(CitGVD,http://citgvd.cric.cn/home)was developed as the first citrusspecific comprehensive database dedicated to genome-wide variations including single nucleotide polymorphisms(SNPs)and insertions/deletions(INDELs).The current version(V1.0.0)of CitGVD is an open-access resource centered on 1,493,258,964 high-quality genomic variations and 84 phenotypes of 346 organisms curated from in-house projects and public resources.CitGVD integrates closely related information on genomic variation annotations,related gene annotations,and details regarding the organisms,incorporating a variety of built-in tools for data accession and analysis.As an example,CitGWAS can be used for genome-wide association studies(GWASs)with SNPs and phenotypic data,while CitEVOL can be used for genetic structure analysis.These features make CitGVD a comprehensive web portal and bioinformatics platform for citrus-related studies.It also provides a model for analyzing genome-wide variations for a wide range of crop varieties.

CsPrx25,a class III peroxidase in Citrus sinensis,confers resistance to citrus bacterial canker through the maintenance of ROS homeostasis and cell wall lignification

Citrus bacterial canker(CBC)results from Xanthomonas citri subsp.citri(Xcc)infection and poses a grave threat to citrus production.Class III peroxidases(CIII Prxs)are key proteins to the environmental adaptation of citrus plants to a range of exogenous pathogens,but the role of CIII Prxs during plant resistance to CBC is poorly defined.Herein,we explored the role of CsPrx25 and its contribution to plant defenses in molecular detail.Based on the expression analysis,CsPrx25 was identified as an apoplast-localized protein that is differentially regulated by Xcc infection,salicylic acid,and methyl jasmone acid in the CBC-susceptible variety Wanjincheng(C.sinensis)and the CBC-resistant variety Calamondin(C.madurensis).Transgenic Wanjincheng plants overexpressing CsPrx25 were generated,and these transgenic plants exhibited significantly increased CBC resistance compared with the WT plants.In addition,the CsPrx25-overexpressing plants displayed altered reactive oxygen species(ROS)homeostasis accompanied by enhanced H_(2)O_(2) levels,which led to stronger hypersensitivity responses during Xcc infection.Moreover,the overexpression of CsPrx25 enhanced lignification as an apoplastic barrier for Xcc infection.Taken together,the results highlight how CsPrx25-mediated ROS homeostasis reconstruction and cell wall lignification can enhance the resistance of sweet orange to CBC.

CsWAKL08,a pathogen-induced wall-associated receptor-like kinase in sweet orange,confers resistance to citrus bacterial canker via ROS control and JA signaling

Citrus bacterial canker(CBC)is a disease resulting from Xanthomonas citri subsp.citri(Xcc)infection and poses a grave threat to citrus production worldwide.Wall-associated receptor-like kinases(WAKLs)are proteins with a central role in resisting a range of fungal and bacterial diseases.The roles of WAKLs in the context of CBC resistance,however,remain unclear.Here,we explored the role of CsWAKL08,which confers resistance to CBC,and we additionally analyzed the molecular mechanisms of CsWAKL08-mediated CBC resistance.Based on systematic annotation and induced expression analysis of the CsWAKL family in Citrus sinensis,CsWAKL08 was identified as a candidate that can be upregulated by Xcc infection in the CBC-resistant variety.CsWAKL08 can also be induced by the phytohormones salicylic acid(SA)and methyl jasmonic acid(MeJA)and spans the plasma membrane.Overexpression of CsWAKL08 resulted in strong CBC resistance in transgenic sweet oranges,whereas silencing of CsWAKL08 resulted in susceptibility to CBC.The peroxidase(POD)and superoxide dismutase(SOD)activities were significantly enhanced in the CsWAKL08-overexpressing plants compared to the control plants,thereby mediating reactive oxygen species(ROS)homeostasis in the transgenic plants.Moreover,the JA levels and the expression of JA biosynthesis and JA responsive genes were substantially elevated in the CsWAKL08 overexpression plants relative to the controls upon Xcc infection.Based on these findings,we conclude that the wall-associated receptor-like kinase CsWAKL08 positively regulates CBC resistance through a mechanism involving ROS control and JA signaling.These results further highlight the importance of this kinase family in plant pathogen resistance.

Pathologically expanded peripheral CD4^(+)PD-1^(+)Foxp3^(−) T-cell subset promotes B-cell hyperactivity in patients with rheumatoid arthritis

Background:Rheumatoid arthritis (RA) is an autoimmune disease characterized by destructive polyarthritis, and abnormal T-B-cell interactions may contribute to its pathogenesis. This study aimed to investigate the characteristics and roles of CD4^(+) programmed death 1 (PD-1)^(+)Foxp3^(−) T cells in relation to the B-cell response in patients with RA. Methods:This study included 155 patients with RA and 36 age- and sex-matched healthy controls (HCs) from the Second Hospital of Dalian Medical University in China. Flow cytometry was used to assess the proportion and properties of peripheral CD4^PD-1^(−)Foxp3^(+) T cells, including their proliferation, activation, cytokine production, and capacity to induce B-cell differentiation. Results:The proportion of CD4^(+)PD-1^(+)Foxp3^(−)T cells was increased in patients with RA compared with HCs ([10.78 ± 0.60]% vs. [5.67 ± 0.40]%, p < 0.001), and this was positively associated with the B-cell response. Compared with CD4^PD-1 ^(+)Foxp3 ^(+) T cells, CD4^(+)PD-1^(+)Foxp3^(−)T cells from patients with RA exhibited increased expression of Ki67 ([6.52 ± 0.41]% vs. [3.87 ± 0.42]%, p < 0.01) and activation markers, produced higher levels of cytokines, and showed enhanced B-cell differentiation. Furthermore, anti-interleukin-6R antagonists decreased the proportion, activation, and cytokine production of CD4^(+)PD-1^(+)Foxp3^(−)T cells in vitro. The frequency of type 2 CD4^(+)PD-1^(+)Foxp3^(−)T cells was significantly higher in patients with RA than that in HCs ([37.27 ± 1.43]% vs. [29.05 ± 1.30]%, p < 0.05). Conclusions:Peripherally expanded CD4^(+)PD-1^(+)Foxp3^(−)T cells in patients with RA, which induced B-cell hyperactivity, may be inclined toward type 2 helper T cells. Our findings revealed a novel T-cell subset that contributes to B-cell hyperactivity in the pathogenesis of RA.

Identification of two migratory colon ILC2 populations differentially expressing IL-17A and IL-5/IL-13

Group 2 innate lymphoid cells(ILC2s)play important tissue resident roles in anti-parasite immunity,allergic immune response,tissue homeostasis,and tumor immunity.ILC2s are considered tissue resident cells with little proliferation at steady state.Recent studies have shown that a subset of small intestinal ILC2s could leave their residing tissues,circulate and migrate to different organs,including lung,liver,mesenteric LN and spleen,upon activation.However,it remains unknown whether other ILC populations with migratory behavior exist.In this study,we find two major colon ILC2 populations with potential to migrate to the lung in response to IL-25 stimulation.One subset expresses IL-17A and resembles inflammatory ILC2s(iILC2s)but lacks CD27 expression,whereas the other expresses CD27 but not IL-17A.In addition,the IL-17A^(+)ILC2s express lower levels of CD127,CD25,and ST2 than CD27^(+)ILC2s,which express higher levels of IL-5 and IL-13.Surprisingly,we found that both colon ILC2 populations still maintained their colonic features of preferential expression of IL-17A and CD27,IL-5/IL-13,respectively.Together,our study identifies two migratory colon ILC2 subsets with unique surface markers and cytokine profiles which are critical in regulating lung and colon immunity and homeostasis.

基于脂蛋白P80的滑液支原体抗体ELISA检测方法的建立及应用

【目的】研究滑液支原体(Mycoplasma synoviae, MS)脂蛋白P80的免疫反应性及其在MS血清抗体ELISA检测中的应用。【方法】对MSP80的氨基酸序列进行生物信息学分析、原核表达和纯化,并用免疫印迹法分析其与6种不同MS分离株阳性血清的免疫反应性以及与其他禽病原血清的交叉反应性;运用纯化的MS P80表达蛋白作为包被抗原建立了MS血清抗体的间接ELISA检测方法,对其敏感性和重复性进行检测;比较检测了与美国爱德士检测试剂盒对50份临床血清样品的阳性符合率。【结果】生物信息学分析预测MS P80蛋白为脂蛋白且含有信号肽,其在MS种内同源性高达98%-100%,与其他种属P80蛋白同源性在25%-34%之间,成功表达和纯化了MS P80重组蛋白(rMS P80);Western blotting分析表明纯化的rMS P80具有良好的免疫反应性和特异性;运用rMS P80建立的MS血清ELISA抗体检测方法可对不同株MS阳性血清进行抗体效价检测,而对其他禽病原阳性血清均无交叉反应性;该检测方法的批内变异系数小于5%,批间变异系数小于10%,重复性良好;与美国IDEXX检测试剂盒比较,本文建立的ELISA抗体检测方法敏感性更高,阳性符合率为75%,阴性符合率为89.47%,总样本符合率为86%。【结论】MS P80具有较好的免疫反应性、种内保守性和种间特异,并且可用作MS抗体检测的靶标抗原。

Clerodenoids A-F:C-ring Aromatized and/or Rearranged Abietane Diterpenoids from Clerodendrum chinense var.simplex

Main observation and conclusion As part of our continuing efforts toward the discovery of the biologically active diterpenoids from medicinal plants,six new C-ring aromatized abietane diterpenoids,clerodenoids A-F(1-6),were isolated from Clerodendrum chinense.

Engineering Non-transgenic Gynoecious Cucumber Using an Improved Transformation Protocol and Optimized CRISPR/Cas9 System

Dear Editor,Gynoecism has been extensively exploited in cucumber breeding. The utilization of a gynoecious line permits earlier pro- duction of hybrids, higher yield, and more concentrated fruit set. In addition, the utilization of a gynoecious line eliminates the need for hand emasculation and reduces the labor cost of crossing （Robinson, 2000）. Therefore, the development of gynoeoious inbred lines is instrumental for cucumber breeding. Gynoecious inbreds can be produced by selection from crosses of monoecious inbreds, or can arise spontaneously from natural variation. However, both methods have disadvantages. For instance, the time-consuming and laborious process of crossing can also lead to the introduction of undesirable traits,

Developmentally Regulated Glucosylation of Bitter Triterpenoid in Cucumber by the UDP-Glucosyltransferase UGT73AM3

Dear Editor,Plants have evolved great plasticity to adapt to external environments. A huge number of structurally diverse metabolites gener- ated through the glycosylation process is one potential mechanism that contributes to this plasticity （Bowles et al., 2005）.

Preparation of ultrathin ReS2 nanosheets and their application to Q-switched Er-doped fiber lasers

We study the exfoliation of ultrathin ReS2 nanosheets from the prepared ReS2 powder and their application to Q-switched Er-doped fiber laser.XRD,Raman,and XPS spectra confirm the successful preparation of the layered ReS2.SEM images show that the obtained ReS2 sheets have lateral size below 200 nm.The thickness of the ReS2 nanosheets is smaller than 5 nm according to the AFM results.ReS2/PVA film is applied as a saturable absorber in an Er-doped Q-switched fiber laser,and a minimum pulse duration of 2.4μs and an output power of 1.25 mW are obtained,indicating the potential application to Q-switched lasers.