Molecular regulation of oil gland development and biosynthesis of essential oils in Citrus spp.

Secretory structures in terrestrial plants serve as reservoirs for a variety of secondary metabolites. Among these, the secretory cavity of the Rutaceae family is notable for containing essential oils with a wide range of applications. However, the molecular basis underlying secretory cavity development is unknown. Here, we reveal a molecular framework for Citrus oil gland formation. Using genetic mapping and genome editing, we demonstrated that this process requires LATE MERISTEM IDENTITY1 (LMI1), a key regulator of leaf serration. A conserved GCC box element of the LMI1 promoter recruits DORNROSCHEN-like (DRNL) for transcriptional activation. This DRNL-LMI1 cascade triggers MYC5 activation, facilitating the development of oil glands and the biosynthesis of essential oils. Our findings spotlight cis-regulatory divergence within leaf shape genes, propelling novel functional tissue formation.

Ethylene activation of carotenoid biosynthesis by a novel transcription factor CsERF061.

Chromoplast-specific lycopene ß-cyclase (LCYb2) is a critical carotenogenic enzyme, which controls the massive accumulation of downstream carotenoids, especially provitamin A carotenoids, in citrus. Its regulatory metabolism is largely unknown. Here, we identified a group I ethylene response factor, CsERF061, in citrus by yeast one-hybrid screen with the promoter of LCYb2. The expression of CsERF061 was induced by ethylene. Transcript and protein levels of CsERF061 were increased during fruit development and coloration. CsERF061 is a nucleus-localized transcriptional activator, which directly binds to the promoter of LCYb2 and activates its expression. Overexpression of CsERF061 in citrus calli and tomato fruits enhanced carotenoid accumulation by increasing the expression of key carotenoid pathway genes, and increased the number of chromoplasts needed to sequester the elevated concentrations of carotenoids, which was accompanied by changes in the concentrations of abscisic acid and gibberellin. Electrophoretic mobility shift and dual-luciferase assays verified that CsERF061 activates the promoters of nine other key carotenoid pathway genes, PSY1, PDS, CRTISO, LCYb1, BCH, ZEP, NCED3, CCD1, and CCD4, revealing the multitargeted regulation of CsERF061. Collectively, our findings decipher a novel regulatory network of carotenoid enhancement by CsERF061, induced by ethylene, which will be useful for manipulating carotenoid accumulation in citrus and other plants.

Value of electron microscopy in the pathological diagnosis of native kidney biopsies in children.

BACKGROUND: Pediatric native kidney diseases are common worldwide. The pathological diagnosis of kidney lesions is crucial for clinical treatment and prognosis. The aim of the current study was therefore to evaluate the value of electron microscopy (EM) to the final diagnosis of native kidney biopsies in children. METHODS: A retrospective evaluation of 855 pediatric kidney biopsies obtained from the Department of Pediatrics in Peking University First Hospital between November 2010 and December 2017 was performed to assess the contribution of EM to the final diagnosis. RESULTS: The role of EM in the final diagnosis was determined to be crucial in 300 cases (35.1%), important in 280 cases (32.7%), and auxiliary in 275 cases (32.2%). EM is considered most valuable in a large percentage of glomerular diseases, mainly including minimal change disease, early-stage membranous nephropathy, postinfectious glomerulonephritis, Alport syndrome, thin basement membrane nephropathy, and thrombotic microangiopathy. EM also provided helpful diagnostic information in cases of focal segmental glomerulosclerosis, lupus nephritis, IgA nephropathy, and IgA vasculitis (Henoch-Schonlein purpura nephritis). Additionally, EM was crucial in 90.0% of cases of subtle pathological changes observed with light microscopy (LM) and immunofluorescence (IF) and in 69.3% of the IF-negative specimens. Patients with nephrotic syndrome or hematuria also benefit from ultrastructural examination. CONCLUSIONS: The present study demonstrated the crucial or important role of EM in the diagnosis of a majority of native kidney biopsies in children. The application of EM should be integrated together with LM and IF as a routine method of assessing pediatric kidney specimens. Graphical abstract.

The relationship between the expressions of tumor associated fibroblasts Cav-1 and MCT4 and the prognosis of papillary carcinoma of breast.

The aim of this study was to evaluate the expression levels of Cav-1 and MCT4 in the tumor stroma CAFs. Immunohistochemical method was used to detect the expression of Cav-1 and MCT4 in 86 cases of IMPC, with 105 cases of non-specific invasive ductal carcinoma (IDC-NOS) as control. Their differential expression in different histological types were compared and relationship between the expression of Cav-1 and IMPC in the MCT4 and the main pathological features such as age, tumor size and histological grades were analyzed. The study confirmed that the loss expression of Cav-1 on CAFs and the up-regulation of MCT4 may be the possible mechanisms of CAFs in tumorigenesis, which may be one of the molecular mechanisms leading to the poor prognosis of IMPC. The expression of CAFs related proteins in tumor microenvironment suggested that CAFs could be used as the target of tumor therapy and provide new evidence for the treatment of breast cancer.

[Application of serratus anterior muscle flap combined with breast implants for breast reconstruction after modified radical mastectomy].

Objective: To investigate effectiveness of the combination of serratus anterior muscle flap and breast implants for breast reconstruction after modified radical mastectomy. Methods: Between January 2015 and December 2015, 25 female patients with breast cancer were enrolled, aged 24-62 years (mean, 40.6 years). The tumor located at left side in 9 cases and right side in 16 cases; 14 cases were in the left upper quadrant, 4 cases were in the left lower quadrant, 7 cases were on the top of the breast. All cases were invasive ductal carcinoma. According to TNM staging, 14 cases were at stageⅠand 11 cases were at stageⅡA. The diameter of lumps were all less than 3 cm. All those lumps were solitary and without distant metastasis. The sentinel nodes were all negative. After modified radical mastectomy, the breasts were reconstructed by serratus anterior muscle flap and breast implants. The nipples were spared in 22 cases. Results: The operation time was 113-148 minutes (mean, 136 minutes). All breasts survived and incisions healed at stageⅠ. There was no complication such as hematoma, infection, etc. All patients were followed up 6-18 months (mean, 15 months). Except 1 case, the others were evaluated according to the criteria of the reconstructed breast at 12 months after operation. Among them, 23 cases were evaluated as good and 1 case as fair. There was no tumor recurrence during the follow-up period. Conclusion: The combination of serratus anterior muscle flap and breast implants after the modified radical mastectomy is a handy approach of breast reconstruction which is less harmful with few postoperative complications. It also gains a high degree of satisfaction from patients for good breast shape.

Autophagy modification augmented the treatment effects initiated by arsenic trioxide in NB4 cells.

Autophagy is a survival pathway required for cellular viability during starvation through catabolic self-digestion of damaged proteins and organelles; however, autophagy may result in cell death if it proceeds to completion. Although the exact mechanism of this process is not clear, it seems that proper regulation of autophagy can potentially contribute to the therapeutics of cancers. This study was designed to examine the role of autophagy in the death of human acute promyelocytic leukemia NB4 cells initiated by arsenic trioxide. Furthermore, the effects of autophagy inhibition and augmentation on cell viability were also compared. Our data suggested that both augmentation and suppression of autophagy could enhance the treatment effects while the latter was preferable. This study indicated that autophagy regulation augmented the treatment effects initiated by arsenic trioxide in NB4 cells, and that the selection of regulator should be precisely considered.

Hydrogen sulfide regulates cardiac function and structure in adriamycin-induced cardiomyopathy.

BACKGROUND: The present study was designed to investigate if hydrogen sulfide (H2S), a novel gasotransmitter, might have a regulatory effect on cardiac function and structure, as well as oxidative stress, in adriamycin (ADR)-induced cardiomyopathy. METHODS AND RESULTS: Hemodynamic measurements, histopathological examination and stereological ultrastructural analysis of mitochondria in ADR-treated rats showed characteristics of cardiomyopathy with remarkable greater size and smaller number of cardiomyocytic mitochondria and a significantly low H2S content in plasma and myocardium, but increased levels of thiobarbituric acid reactive substance (TBARs) and decreased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in plasma and myocardium compared with controls (P<0.01). However, administration of the H2S donor, NaHS, markedly improved cardiac function, as demonstrated by elevated left ventricular developed pressure (+/-LVdp/dtmax; P<0.01) with ameliorated morphological alterations in the myocardium. Myocardial TBARs content decreased, whereas the activities of SOD and GSH-Px increased (P<0.01 and P<0.05, respectively). CONCLUSIONS: Downregulation of endogenously-generated H2S is probably involved in the pathogenesis of ADR-induced cardiomyopathy, whereby H2S reduces lipid peroxidation, increases antioxidation, and inhibits oxidative stress injury.

[Ultrastructural and immunoelectron microscopic study on light chain deposition diseases and primary amyloidosis of the kidney].

OBJECTIVE: To investigate the role of electron microscopy (EM) and immunoelectron microscopy (IEM) in the diagnosis of light chain deposition diseases (LCDD) and primary amyloidosis (AL) of the kidney. METHODS Renal biopsy specimens collected in our hospital from Jan. 1994 to Feb. 2002 were studied by EM, and ultrastructural immunogold labeling for light chains (kappa, Lambda) were made on the cases suggested of LCDD and early stage AL in the preliminary diagnosis. RESULTS: Six cases were diagnosed as LCDD, and punctate, granular, electron-dense material was identified in the outside of tubular basement membrane (TBM), inner layer of glomerular basement membrane (GBM) and vascular walls by EM. Two of which showed interrupted segmental localization of granular material in subendothelial areas of GBM. The granular material was labeled by monotypical light chains (4 cases of kappa chain, 2 cases of Lambda chain). Thirty-six cases were diagnosed as AL. EM revealed fibrillary material distributed in glomerular mesangium, GBM and vascular walls; 15 of them showed segmental distribution of fibrils in mesangium and/or GBM, and were further studied by IEM labeling for light chains. The fibrillary material was labeled by monotypical light chains (4 cases of kappa chain, 11 cases of Lambda chain). CONCLUSION The diagnosis of renal LCDD and early stage of AL depended on EM, IEM is a unique diagnostic tool in early stage of LCDD and AL.