Full-Length Transcriptome Sequencing Reveals the Impact of Cold Stress on Alternative Splicing in Quinoa.

Quinoa is a cold-resistant and nutrient-rich crop. To decipher the cold stress response of quinoa, the full-length transcriptomes of the cold-resistant quinoa variety CRQ64 and the cold-sensitive quinoa variety CSQ5 were compared. We identified 55,389 novel isoforms and 6432 novel genes in these transcriptomes. Under cold stress, CRQ64 had more differentially expressed genes (DEGs) and differentially alternative splicing events compared to non-stress conditions than CSQ5. DEGs that were specifically present only in CRQ64 were significantly enriched in processes which contribute to osmoregulation and ROS homeostasis in plants, such as sucrose metabolism and phenylpropanoid biosynthesis. More genes with differential alternative splicing under cold stress were enriched in peroxidase functions in CRQ64. In total, 5988 transcription factors and 2956 long non-coding RNAs (LncRNAs) were detected in this dataset. Many of these had altered expression patterns under cold stress compared to non-stress conditions. Our transcriptome results demonstrate that CRQ64 undergoes a wider stress response than CSQ5 under cold stress. Our results improved the annotation of the quinoa genome and provide new insight into the mechanisms of cold resistance in quinoa.

Casein kinase 1α inhibits p53 downstream of MDM2­mediated autophagy and apoptosis in acute myeloid leukemia.

Enhancement of autophagy serves as a promising therapeutic strategy for cancer, including acute myeloid leukemia (AML). Casein kinase 1α (CK1α), encoded by CSNK1A1, regulates Wnt/ß­catenin, p53 and other key signaling pathways, and is critically involved in tumor progression. However, the relationship and mechanism of CK1α with autophagy in AML still remain unclear. In the present study, it was found that AML patients had higher expression of CSNK1A1 mRNA than healthy donors. Furthermore, we analyzed 163 cases of AML patients in the LAML database of TCGA and found that AML patients with high CSNK1A1 had shorter overall survival than those with low or medium CSNK1A1 expression. Furthermore, we demonstrated that CK1α was a negative regulator of autophagy and apoptosis. Pharmacologic inhibition of CK1α using D4476 or CK1α knockdown via lentivirus­mediated shRNA suppressed proliferation and the clone formation by enhancing autophagic flux and apoptosis in AML cell lines as well as in patient blast cells. Intriguingly, D4476­induced cell death was aggravated in combination with an autophagy inhibitor, Spautin­1, suggesting that autophagy may be a pro­survival signaling. CK1α interacted with murine double minute 2 (MDM2) and p53, and CK1α inhibitor D4476 significantly upregulated p53 and phosphorylated 5' AMP­activated protein kinase (AMPK), and substantially inhibited the phosphorylation of mammalian target of rapamycin (mTOR). Our findings indicate that CK1α promotes AML by suppressing p53 downstream of MDM2­mediated autophagy and apoptosis, suggesting that targeting CK1α provides a therapeutic opportunity to treat AML.

The clinical features of posterior scleritis with serous retinal detachment: a retrospective clinical analysis.

AIM: To summarize the clinical features, systemic associations, risk factors and choroidal thickness (CT) changing in posterior scleritis (PS) with serous retinal detachment. METHODS: This retrospective study included 23 patients diagnosed PS with retinal detachment from August 2012 to July 2017. All patients' medical history and clinical features were recorded. The examinations included best corrected visual acuity (BCVA), intraocular pressure (IOP), fundus examination, and routine eye examinations. Posterior coats thickness (PCT) was determined by B-scan ultrasound, the CT was measured by enhanced depth imaging spectral-domain optical coherence tomography (EDI-OCT) and clinical data were compiled and analyzed. RESULTS: After application of extensive exclusion criteria, 23 patients with PS remained (13 females, 10 males). The average age at presentation was 29.5±9.24 years old. Ocular pain and blurred vision were the two most common complained symptoms by patients. Anterior scleritis occurred in 12 patients, which was confirmed by ultrasound biomicroscopy (UBM) examination. Despite all patients displaying serous retinal detachment in their macula, no fluorescein leakage was observed in the macular area. Optic disc swelling was documented in 10 of the 23 eyes. From B-scan ultrasound examination, the PCT increased with fluid in Tenon's capsule demonstrated as a typical T-sign. The average PCT was 2.51±0.81 mm in the PS-affected eyes and only 1.09±0.29 mm in the unaffected eye (P<0.0001). The subfoveal CT was 442.61±55.61 µm, which correlated with axis length (r=-0.65, P=0.001) and PCT (r=0.783, P<0.001). The BCVA and IOP did not correlate with either CT or PCT. CONCLUSION: PS with serous retinal detachment presented a variety of symptoms, such as pain, visual loss, and physical indicators. Typical T-sign detected by B-scan ultrasound is a useful confirmatory sign for PS diagnosis. Pathological increases in CT might be a potential predictive factor for inflammation.

Amyloid Beta Deposition Could Cause Corneal Epithelial Cell Degeneration Associated with Increasing Apoptosis in APPswePS1 Transgenic Mice.

OBJECTIVE: To investigate the expression of amyloid precursor protein (APP) and amyloid beta (Aß) in cornea and further explore the pathological and ultrastructural changes in corneal epithelium in APPswePS1 transgenic mice. METHODS: Twelve wild type mice were grouped into control group and twelve TgAPPswePS1 mice at least 8 months old were grouped into the young experiment group (Tg-8M group), and another twelve transgenic mice at least 15 months old were selected into the aged experiment group (Tg-15M group). The pathological degeneration, ultrastructural changes, and the expression of APP, Aß deposition, and the TUNEL reaction in corneal epithelial cells were observed. Western blot analysis was performed to determine expression levels of APP and Aß with scraped epithelial debridement. All the results were quantified and analyzed. RESULTS: In transgenic mice, the H&E-stained cornea sections demonstrated histopathological changes in corneal epithelial cells with irregular arrangement and the number of cell layers decreased, while normal structure observed in controls. In Tg-15M group, the corneal epithelial cell displaced a significant number of intracellular vacuoles with 1-2 cell layers left. Transmission electron microscopy (TEM) further confirmed the dramatic degeneration in corneal epithelium, the microvilli suffered degenerative changes and found with typical fingerpoint-like morphology in controls; however, microspike-like in Tg-15M group, and the number of microvilli decreased considerabely. An APP-positive immunoreaction was detected with a diffuse pattern in the corneal epithelial cells layer, about 3.122 ± 0.596 and 7.372 ± 0.936 fold changes in Tg-8M and Tg-15M groups, respectively, as compared with controls. On corneal flatmount, Aß deposition found a diffuse pattern in the cytoplasm by fluorescence staining in TgAPPswePS1 with significantly increasing as compared with the controls, but no plaque was found. The apoptosis of TUNEL cells were observed in TgAPPswePS1 mice and increased 16.329 ± 3.542 fold changes in Tg-15M group as compared with controls. CONCLUSION: The APP expression and Aß deposition might cause cornea epithelial cells degeneration in TgAPPswePS1 mice, associated with apoptosis in basal lamina cells.

Amyloid beta deposition related retinal pigment epithelium cell impairment and subretinal microglia activation in aged APPswePS1 transgenic mice.

AIM: To identify the pathological role of amyloid beta (Aß) deposition in retinal degeneration, and explore Aß deposition on the retinal pigment epithelium cells (RPE) layer and the associated structural and functional changes in Alzheimer's disease transgenic mice. METHODS: RPE changes in the eyes of APPswe/PS1 transgenic and none transgenic (NTG) mice over 20 months old were examined. Histological changes were investigated via hematoxylin and eosin (H&E) staining and transmission electron microscopy (TEM) examination, whereas the expression of amyloid precursor protein (APP), Aß, Zonula occludens-1 (ZO-1) and Ionized calcium binding adaptor molecule-1 (IBA-1) were investigated using immunohistochemistry and immunofluorescence techniques. All of the obtained results were quantitatively and statistically analyzed. RESULTS: In aged transgenic mice, an APP-positive immunoreaction and Aß deposition were detected on the RPE layer but were undetectable in NTG mice. The RPE demonstrated some vacuole changes, shortened basal infoldings and basal deposition in histopathological examination and TEM tests, wherein irregular shapes were indicated by ZO-1 disorganization through fluorescence. Furthermore, IBA-1 positive cells were observed to have accumulated and infiltrated into the RPE layer and localized beneath the RPE/Bruch's membrane (BrM) complex, which was accompanied by an increase in BrM thickness in aged transgenic mice in comparison to NTG mice. The IBA-1 positive cells were found to be co-stained with Aß deposition on the RPE flat mounts. CONCLUSION: The observed Aß deposition in the RPE layer may cause RPE dysfunction, which is associated with microglia cells infiltration into the retina of aged transgenic mice, suggesting that Aß deposition probably plays a significant role in RPE-related degenerative disease.

Short-Term Deposition of PM2.5 Particles on Contact Lens Surfaces: Effect on Oxygen Permeability and Refractive Index.

PURPOSES: To identify the deposition of fine (≤2.5 µm diameter) particulate matter (PM) particles (PM2.5) on contact lens surfaces and to investigate the effects of such deposition on the oxygen permeability (OP) and refractive index (RI) of contact lenses. METHODS: A total of 36 contact lenses, including rigid gas permeable (RGP) lens and soft contact lens (SCL), were investigated. RGP lens (n=12) and SCL (n=12) (experimental group) were incubated in a PM2.5 solution for 24 h, after which PM2.5-treated RGP lens (n=6) and SCL (n=6) were further washed for 1 h in phosphate-buffered saline (PBS). All lenses were examined by field emission scanning electron microscopy. OP and RI of all lenses were measured. RESULTS: Average-sized PM2.5 particles deposited on RGP contact lens and SCL surfaces after immersion in the PM2.5 solution were 3.192 ± 1.637 and 2.158 ± 1.187/100 µm2, respectively. On RGP lens surfaces, we observed both large (≥2.5 µm diameter) and small (PM2.5) particles. PM2.5 particles were deposited in diffuse patterns, primarily along the honeycomb structural border of SCL, while no PM2.5 particles were found in the honeycomb hole of SCL surfaces. Washing in PBS removed the larger PM particles from RGP lens surfaces, but left copious amounts of PM2.5 particles. In contrast, nearly all PM particles were removed from SCL surfaces after PBS washing. OP values of RGP lens and SCL appeared to be unchanged by PM2.5 deposition. RI values increased in both RGP lens and SCL groups after PM2.5 deposition. However, these increases were not statistically significant, suggesting that PM2.5 deposition itself does not cause fluctuations in contact lens RI. CONCLUSIONS: Deposition of PM2.5 particles on contact lens surfaces varies according to lens material. PM2.5 particles deposited on SCL, but only large particles on RGP surfaces were able to be removed by washing in PBS and did not appear to alter OP and RI of either lens type.

[Effect of ABO-Incompatibility on Outcome of Allogeneic Hematopoietic Stem Cell Transplantation].

OBJECTIVE: To investigate the effect of ABO-incompatibility on the efficacy and complications of allogeneic hematopoietic stem cell transplantation(HSCT). METHODS: The clinical data of 54 recipients who received ABO-incompatible allo-HSCT were retrospectively analyzed and were compared with 54 ABO-identical recipients as controls. Hematopoietic reconstruction and the blood type conversion time were dynamically observed and compared between 2 groups. RESULTS: The time of erythrocyte reconstitution was prolonged to 24 d in ABO-incompatible group, compared with that of 19 d in ABO-compatible group (P<0.01). The difference of neutrophil and platelet reconstruction was not statistically significant (P>0.05). Major mismatch group and bidirectional mismatch group required more erythrocyte transfusions than that of ABO-compatible group. The surface antigen of erythrocyte change in major mismatch group was earlier than that of minor mismatch group (P<0.05). The incidence of cytomegalovirus (CMV) infection, acute graft versus host disease (aGVHD) and survival were not significantly different between 2 groups. CONCLUSION: ABO-incompatibility can not influence the effect of allo-HSCT, but ABO-incompatibility delayed erythrocyte recovery, and required more RBC and platelet transfusions.