It is well known that phosphorylation of extracellular signal‐regulated kinase (ERK) is involved in prothoracicotropic hormone (PTTH)‐stimulated ecdysteroidogenesis in insect prothoracic glands ...(PGs). In the present study, we further investigated the downstream signalling pathways. Our results showed that PTTH stimulated p90 ribosomal S6 kinase (RSK) phosphorylation at Thr573 in Bombyx mori PGs both in vitro and in vivo. The in vitro PTTH stimulation was stage‐ and dose‐dependent. The absence of Ca2+ reduced PTTH‐stimulated RSK phosphorylation. Stimulation of RSK phosphorylation was also observed after treatment with either A23187 or thapsigargin. A phospholipase C (PLC) inhibitor, U73122, blocked PTTH‐stimulated RSK phosphorylation. These results indicate the involvement of Ca2+ and PLC. Treatment with diphenylene iodonium (DPI), a mitochondrial oxidative phosphorylation inhibitor, blocked PTTH‐regulated RSK phosphorylation, indicating its redox regulation. A mitogen‐activated protein kinase (MAPK)/ERK kinase (MEK) inhibitor, U0126, but not a phosphatidylinositol 3‐kinase (PI3K) inhibitor, LY294002, decreased PTTH‐stimulated RSK phosphorylation, indicating that ERK is an upstream signalling. A protein kinase C (PKC) inhibitor, chelerythrine C, inhibited PTTH‐stimulated RSK phosphorylation, and a PKC activator, phorbol 12‐myristate acetate (PMA) stimulated RSK phosphorylation, indicating the involvement of PKC. BI‐D1870, a specific RSK inhibitor, partly prevented PTTH‐stimulated RSK phosphorylation and significantly inhibited PTTH‐stimulated ecdysteroid secretion, indicating that PTTH‐stimulated RSK phosphorylation is involved in ecdysteroidogenesis. Taken together, these data indicate that PTTH activates RSK phosphorylation which plays important roles in PTTH‐stimulated ecdysteroidogenesis.
PTTH stimulated RSK phosphorylation in Bombyx PGs both in vitro and in vivo.
MEK inhibitor U0126 decreased PTTH‐stimulated RSK phosphorylation.
The complex signalling involved PTTH‐stimulated RSK phosphorylation was investigated.
RSK appears to be involved in PTTH‐stimulated ecdysteroidogenesis.
Summary
Background
Pioneering effort has been made to facilitate the recognition of pathology in malignancies based on whole‐slide images (WSIs) through deep learning approaches. It remains unclear ...whether we can accurately detect and locate basal cell carcinoma (BCC) using smartphone‐captured images.
Objectives
To develop deep neural network frameworks for accurate BCC recognition and segmentation based on smartphone‐captured microscopic ocular images (MOIs).
Methods
We collected a total of 8046 MOIs, 6610 of which had binary classification labels and the other 1436 had pixelwise annotations. Meanwhile, 128 WSIs were collected for comparison. Two deep learning frameworks were created. The ‘cascade’ framework had a classification model for identifying hard cases (images with low prediction confidence) and a segmentation model for further in‐depth analysis of the hard cases. The ‘segmentation’ framework directly segmented and classified all images. Sensitivity, specificity and area under the curve (AUC) were used to evaluate the overall performance of BCC recognition.
Results
The MOI‐ and WSI‐based models achieved comparable AUCs around 0·95. The ‘cascade’ framework achieved 0·93 sensitivity and 0·91 specificity. The ‘segmentation’ framework was more accurate but required more computational resources, achieving 0·97 sensitivity, 0·94 specificity and 0·987 AUC. The runtime of the ‘segmentation’ framework was 15·3 ± 3·9 s per image, whereas the ‘cascade’ framework took 4·1 ± 1·4 s. Additionally, the ‘segmentation’ framework achieved 0·863 mean intersection over union.
Conclusions
Based on the accessible MOIs via smartphone photography, we developed two deep learning frameworks for recognizing BCC pathology with high sensitivity and specificity. This work opens a new avenue for automatic BCC diagnosis in different clinical scenarios.
What's already known about this topic?
The diagnosis of basal cell carcinoma (BCC) is labour intensive due to the large number of images to be examined, especially when consecutive slide reading is needed in Mohs surgery.
Deep learning approaches have demonstrated promising results on pathological image‐related diagnostic tasks.
Previous studies have focused on whole‐slide images (WSIs) and leveraged classification on image patches for detecting and localizing breast cancer metastases.
What does this study add?
Instead of WSIs, microscopic ocular images (MOIs) photographed from microscope eyepieces using smartphone cameras were used to develop neural network models for recognizing BCC automatically.
The MOI‐ and WSI‐based models achieved comparable areas under the curve around 0·95.
Two deep learning frameworks for recognizing BCC pathology were developed with high sensitivity and specificity.
Recognizing BCC through a smartphone could be considered a future clinical choice.
Linked Comment: Lee and Soyer. Br J Dermatol 2020; 182:540–541.
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This paper evaluates the efficacy and safety of repeat hepatic resection and radiofrequency ablation in the treatment of recurrent hepatocellular carcinoma
We retrieved and collected all relevant ...articles from the inception to 8 March 2020. After data extraction, we conducted meta-analysis and carried out the heterogeneity test, sensitivity analysis, and publication bias test to evaluate reliability
A total of 12 studies with 1746 patients (rHR 837, RFA 909) were included. rHR was similar to RFA in a one-year overall survival rate (OS), while rHR was superior to RFA in 3- and 5-year OS and 1-, 3-, and 5-year disease-free survival rates (DFS), but the procedure-related complications of RFA were significantly less than those of rHR. Among the subgroups with Milan criteria, rHR was similar to RFA in 1-, 3-, and 5-year OS and 1-year DFS, but superior to RFA in 3- and 5-year DFS.
RFA is the first choice for recurrent HCC meeting Milan criteria. When it does not meet the Milan criteria, minimally invasive treatment should not be carried out at the cost of survival, and rHR should be the first choice.
Odorant binding proteins (OBPs) and chemosensory proteins (CSPs) play essential roles in insect chemosensory recognition. Here, we identified nine OBPs and nine CSPs from the Myzus persicae ...transcriptome and genome. Genomic structure analysis showed that the number and length of the introns are much higher, and this appears to be a unique feature of aphid OBP genes. Three M. persicae OBP genes (OBP3/7/8) as well as CSP1/4/6, CSP2/9 and CSP5/8 are tandem arrayed in the genome. Phylogenetic analyses of five different aphid species suggest that aphid OBPs and CSPs are conserved in single copy across all aphids (with occasional losses), indicating that each OBP and CSP class evolved from a single gene in the common ancestor of aphids without subsequent duplication. Motif pattern analysis revealed that aphid OBP and CSP motifs are highly conserved, and this could suggest the conserved functions of aphid OBPs and CSPs. Three OBPs (MperOBP6/7/10) are expressed antennae specifically, and five OBPs (MperOBP2/4/5/8/9) are expressed antennae enriched, consistent with their putative olfactory roles. M. persicae CSPs showed much broader expression profiles in nonsensory organs than OBPs. None of the nine MperCSPs were found to be antennae specific, but five of them (MperCSP1/2/4/5/6) showed higher expression levels in the legs than in other tissues. MperCSP10 mainly expressed in the antennae and legs. The broad and diverse expression patterns of M. persicae CSPs suggest their multifunctions in olfactory perception, development and other processes.
Epithelial-mesenchymal transition (EMT) is an essential component of metastasis. Our previous study demonstrated that cancer-associated fibroblasts (CAFs) induce EMT in lung cancer cells. In recent ...years, many studies have demonstrated that CAFs induce metastasis and drug resistance in cancer cells via exosomes.
We sought to discover the mechanism underlying how CAFs induce EMT in lung cancer cells, unveiling the role of exosomes in lung cancer progression.
We cultured lung cancer cell (i) with control medium, normal fibroblasts (NFs) or CAFs; (ii) with SNAI1-transfected or NC (negative control)-transfected CAFs; (iii) with exosomes extracted from NF- or CAF-conditioned medium; (iv) with exosomes released by SNAI1 or NC-transfected CAFs; (v) with CAF-conditioned medium or exosome-depleted CAF-conditioned medium.
qRT-PCR was conducted to examine the expression of CDH1 (gene of E-cadherin) and VIM (gene of Vimentin), western blotting was conducted to examine E-cadherin and vimentin levels in lung cancer cells.
Exosomes released by CAFs-promoted EMT in lung cancer cells. Interestingly, SNAI1 levels in exosomes secreted from CAFs were correlated with SNAI1 expression in CAFs. Furthermore, the level of SNAI1 in exosomes was crucial for inducing EMT in lung cancer cells. Finally, treatment of CAFs with GW4869, an inhibitor of exosome release, noticeably inhibited their EMT-inducing effect on recipient epithelial cells.
The molecular mechanism underlying how CAFs induce EMT in cancer cells may be that CAFs deliver SNAI1 to recipient cancer cells via exosomes.
Ovarian cancer, the most deadly gynecologic malignancy, is often diagnosed late and at the advanced stage when the cancer cells have already migrated and invaded into other tissues and organs. Better ...understanding of the mechanism of metastasis in ovarian cancer cells is essential to the design of effective therapy. In this study, we investigated the function of scaffolding adaptor protein Gab2 in ovarian cancer cells. Gab2 is found to be overexpressed in a subset of ovarian tumors and cancer cell lines. Gab2 expression mainly regulates the migratory behaviors of ovarian cancer cells. Overexpression of Gab2 promotes the migration and invasion, and downregulates E-cadherin expression in ovarian cancer cells with low-Gab2 expression. Conversely, knockdown of Gab2 expression inhibits the migration and invasion, and promotes E-cadherin expression in ovarian cancer cells with high-Gab2 expression. By expressing Gab2 wild-type and Gab2 mutants that are defective in activation of the PI3K and Shp2-Erk pathways, we find that Gab2 inhibits E-cadherin expression and enhances the expression of Zeb1, a transcription factor involved in epithelial-to-mesenchymal transition (EMT), and cell migration and invasion through the activation of the PI3K pathway. Knockdown of Zeb1 expression blocks Gab2-induced suppression of E-cadherin expression and increase in cell invasion. LY294002 and GDC-0941, inhibitors of PI3K, or Rapamycin, an inhibitor of PI3K downstream target mTOR, can reverse the effects of Gab2 on migration and invasion. Overall, our studies reveal that Gab2 overexpression, via activation of the PI3K-Zeb1 pathway, promotes characteristics of EMT in ovarian cancer cells.
In the present study, the participation of protein kinase C (PKC) signalling in prothoracicotropic hormone (PTTH)‐stimulated ecdysteroidogenesis in Bombyx prothoracic glands (PGs) is demonstrated and ...characterized. PTTH stimulated phosphorylation of a 37‐kDa protein in Bombyx PGs both in vitro and in vivo, as recognized by a PKC substrate antibody. Treatment with either A23187 or thapsigargin also stimulated this 37‐kDa protein phosphorylation. PTTH‐stimulated phosphorylation of the 37‐kDa protein was markedly attenuated in the absence of Ca2+. The phospholipase C (PLC) inhibitor, U73122, greatly inhibited PTTH‐stimulated phosphorylation of this protein, indicating the involvement of Ca2+ and PLC. A mitogen‐activated protein kinase/extracellular signal‐regulated kinase (ERK) kinase (MEK) inhibitor (U0126), a phosphoinositide 3‐kinase (PI3K) inhibitor (LY294002) and a chemical activator of adenosine 5′‐monophosphate‐activated protein kinase (AMPK) (5‐aminoimidazole‐4‐carboxamide‐1‐β‐d‐ribofuranoside) did not affect PTTH‐stimulated phosphorylation of the 37‐kDa protein, implying that ERK and PI3K/AMPK are not the upstream signalling pathways for PKC‐dependent protein phosphorylation. The mitochondrial oxidative phosphorylation inhibitors (the uncoupler carbonyl cyanide p‐trifluoromethoxyphenylhydrazone and diphenylene iodonium) inhibited PTTH‐stimulated phosphorylation of the 37‐kDa protein, indicating its redox regulation. Treatment with PKC inhibitors (either calphostin C, chelerythrine C or rottlerin) reduced PTTH‐stimulated phosphorylation of the 37‐kDa protein. PTTH‐stimulated ecdysteroidogenesis was also inhibited by treatment with rottlerin, thus further confirming participation of PKC‐dependent phosphorylation in PTTH signalling. From these results, we demonstrated that redox‐regulated PTTH‐stimulated PKC signalling is involved in ecdysteroid secretion in Bombyx PGs.
The participation of PKC in Bombyx PTTH signaling was demonstrated.
PTTH stimulated PKC‐dependent phosphorylation of a 37‐kDa protein.
Inhibition of PKC signaling partly decreased PTTH‐stimulated ecdysone secretion.
Oxidative stress injury is one important factor in intestinal mucosal barrier damage. Expression of heat shock protein (HSP)70 is an endogenous mechanism by which living cells adapt to stress. This ...study was undertaken to investigate the protective effects of HSP70 on intestinal oxidative stress. Two hundred and forty broilers were injected intraperitoneally with HSP70 inducer L-(1)-glutamine or with the inhibitor quercetin. Twenty-four hours later, they were heat stressed for 0, 2, 3, 5, and 10 h, respectively, at 36 ± 1°C. The L-(1)-glutamine significantly increased HSP70 expression (P < 0.001). At 2 h or 3 h of heat stress, the HSP70 expression obviously elevated (P < 0.001). Levels of corticosterone and the heterophil:lymphocyte ratio significantly increased when HSP70 expression was inhibited (P < 0.0001). Serum corticosterone was negatively correlated with the HSP70 expression at 3 h of heat stress (P = 0.0015; R = –0.6537). Heat shock protein 70 significantly protected the integrity of the intestinal mucosa from heat stress, with significantly decreased lactic dehydrogenase when HSP70 expression was enhanced (P < 0.001). In addition, heat-stress time significantly affected the lactic dehydrogenase release (P < 0.001). Furthermore, HSP70 significantly elevated antioxidant enzyme activities (such as superoxide dismutase, glutathione peroxidase, and total antioxidant capacity) and inhibited lipid peroxidation to relieve intestinal mucosal oxidative injury (P < 0.001). These results suggest that HSP70 is capable of protecting the intestinal mucosa from heat-stress injury by improving antioxidant capacity of broilers and inhibiting the lipid peroxidation production.
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