Circulating Inflammatory Factor Levels in the Early Phase of COVID-19 are Associated with the Progression of Respiratory Failure: A Single-Center Retrospective Study.

Purpose: To evaluate the potential relationships between serum interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α levels and occurrence of respiratory failure in patients with early-stage COVID-19 disease. Patients and Methods: We analyzed clinical characteristics, laboratory parameters, and immunoinflammatory markers in 302 patients diagnosed with SARS-CoV-2 infection who required hospitalization at Changshu Hospital of Nantong University. IL-2, IL-4, IL-6, IL-10, IL-17, IFN-γ, and TNF-α levels in the peripheral blood of patients hospitalized five days after disease onset were measured using multiplex bead-based flow fluorescent immunoassay (MBFFI). Results: Patients with respiratory failure had higher serum IL-4 [0 (0, 0.54) pg/mL], IL-6 [40.76 (12.33, 90.28) pg/mL], IL-10 [6.65 (4.12, 11.34) pg/mL], and IL-17 [9.48 (4.31, 12.13) pg/mL] levels than patients without respiratory failure (P=0.042, P<0.0001, P=0.012, and P=0.036, respectively). Serum IL-2, IFN-γ, and TNF-α levels were not significantly different between the two groups. The occurrence of respiratory failure was positively correlated with sex (R=0.122, P=0.034), lactic acid (R=0.193, P=0.007), white blood cell count (R=0.121, P=0.038), erythrocyte distribution width (R=0.131, P=0.024), thyrocalcitonin (R=0.280, P<0.0001), and D-dimer levels (R=0.214, P<0.0001) but negatively correlated with oxygen partial pressure (R=-0.208, P=0.004), oxygen saturation (R=-0.220, P=0.002), lymphocyte count (R=-0.129, P=0.026), and calcium (R=-0.152, P=0.042). Among the immunoinflammatory biomarkers, the occurrence of respiratory failure was positively correlated with IL-4 (R=-0.117, P=0.042), IL-6 (R=0.206, P<0.0001), IL-10 (R=0.145, P=0.012), and IL-17 (R=0.121, P=0.036) levels. Conclusion: Serum levels of pro-inflammatory cytokines IL-6 and IL-17 and anti-inflammatory cytokines IL-4 and IL-10 were significantly elevated in patients with respiratory failure and weakly positively correlated with the occurrence of respiratory failure. Further studies are required to explore these key immune mechanisms to help clinicians better manage acute complications, long-term sequelae, and possible future COVID-19 variants and be flexible in managing future epidemics and similar public health threats.

Dysregulation of circulating CD4 + CXCR5 + PD-1+ T cells in diabetic retinopathy.

AIMS: We aimed to determine an association between follicular helper T (Tfh) cells and Bcl-6 and CXCL13 levels and determine the role of Tfh cells, Bcl-6, and CXCL13 serum levels in the pathogenesis of diabetic retinopathy (DR) since Tfh cells have an important role in type 1 diabetes; however, their role in type 2 diabetes-related DR requires exploration. METHODS: Blood samples were collected from 24 patients with non-proliferative diabetic retinopathy (NPDR), 20 with proliferative diabetic retinopathy (PDR), and 18 age- and sex-matched healthy volunteers. Flow cytometry detected CD4 + CXCR5 + PD1+ Tfh cells. Serum Bcl-6 and CXCL13 levels were determined using enzyme-linked immunosorbent assay. RESULTS: CD4 + CXCR5 + PD-1+ Tfh cell percentages in peripheral blood and serum levels of Bcl-6 and CXCL13 in the non-proliferative DR (NPDR) and proliferative DR (PDR) groups' were significantly higher than those in healthy individuals. The proportion of Tfh cells in DR patients' peripheral blood positively correlated with Bcl-6 and CXCL13 serum levels, DR course severity, Fasting blood glucose, glycosylated hemoglobin and body mass index. CONCLUSIONS: The increased circulating Tfh cells, serum Bcl-6 levels, and CXCL13 levels of DR patients with type 2 diabetes suggested that circulating Tfh cells and the germinal center response may have a role in the occurrence and development of DR.

Co-overexpression of AtSAT1 and EcPAPR improves seed nutritional value in maize.

Maize seeds synthesize insufficient levels of the essential amino acid methionine (Met) to support animal and livestock growth. Serine acetyltransferase1 (SAT1) and 3'-phosphoadenosine-5'-phosphosulfate reductase (PAPR) are key control points for sulfur assimilation into Cys and Met biosynthesis. Two high-MET maize lines pRbcS:AtSAT1 and pRbcS:EcPAPR were obtained through metabolic engineering recently, and their total Met was increased by 1.4- and 1.57-fold, respectively, compared to the wild type. The highest Met maize line, pRbcS:AtSAT1-pRbcS:EcPAPR, was created by stacking the two transgenes, causing total Met to increase 2.24-fold. However, the pRbcS:AtSAT1-pRbcS:EcPAPR plants displayed progressively severe defects in plant growth, including early senescence, stunting, and dwarfing, indicating that excessive sulfur assimilation has an adverse effect on plant development. To explore the mechanism of correlation between Met biosynthesis in maize leaves and storage proteins in developing endosperm, the transcriptomes of the sixth leaf at stage V9 and 18 DAP endosperm of pRbcS:AtSAT1, pRbcS:AtSAT1-pRbcS:EcPAPR, and the null segregants were quantified and analyzed. In pRbcS:AtSAT1-pRbcS:EcPAPR, 3274 genes in leaves (1505 up- and 1769 downregulated) and 679 genes in the endosperm (327 up- and 352 downregulated) were differentially expressed. Gene ontology (GO) and KEGG (Kyoto encyclopedia of genes and genomes) analyses revealed that many genes were associated with Met homeostasis, including transcription factors and genes involved in cysteine and Met metabolism, glutathione metabolism, plant hormone signal transduction, and oxidation-reduction. The data from gene network analysis demonstrated that two genes, serine/threonine-protein kinase (CCR3) and heat shock 70 kDa protein (HSP), were localized in the core of the leaves and endosperm regulation networks, respectively. The results of this study provide insights into the diverse mechanisms that underlie the ideal establishment of enhanced Met levels in maize seeds.

Pars Plana Ahmed Valve Implantation for Vitrectomized Eyes With Refractory Glaucoma.

This study aimed to analyze the surgical outcomes of pars plana Ahmed valve implantation in vitrectomized eyes with refractory glaucoma. We performed a retrospective case review of consecutive patients with refractory glaucoma after undergoing pars plana vitrectomy who underwent pars plana Ahmed valve implantation between July 2019 and December 2020 at the glaucoma unit of the Affiliated Changshu Hospital of Xuzhou Medical University (Changshu, China). All the patients were followed up for ≥12 months postoperatively. We recorded pre- to postoperative changes in best-corrected visual acuity (BCVA), intraocular pressure (IOP), number of anti-glaucoma medication, corneal endothelial count, and surgical complications, if any. There was a significant improvement in the median BCVA from 2.30 (0.87, 2.30) logMAR preoperatively to 1.70 (0.70, 2.30) logMAR at discharge and 1.0 (0.52, 1.85) at final examination (p = 0.011, p = 0.001). Compared with the preoperative IOP level, there was a significant decrease in the postoperative IOP at each postoperative time point (p < 0.001). There was a significant reduction in the median number of anti-glaucoma drugs (including postoperative ocular massage), from 3.00 (2.00, 3.00) preoperatively to 0.00 (0.00, 1.00) at the last follow-up postoperative examination (p < 0.001). A 29-year-old woman with proliferative diabetic retinopathy who underwent surgical treatment at 5 months postoperatively for fibrous wrapping formed around the plate of the Ahmed valve showed an IOP of 14 mmHg at the last follow-up. Our findings indicated that pars plana Ahmed valve implantation can be safely performed for managing vitrectomized eyes with refractory glaucoma.

Cyclophotocoagulation under Microscopy Combined with Phacoemulsification for Primary Angle-Closure Glaucoma.

The purpose of this study was to evaluate the safety and efficacy of cyclophotocoagulation under microscopy combined with phacoemulsification in patients with primary chronic angle-closure glaucoma. We retrospectively reviewed the results of cyclophotocoagulation under microscopic direct vision combined with phacoemulsification in 35 eyes (35 patients) with primary chronic angle-closure glaucoma and coexisting visually significant cataracts, treated between January 2017 and April 2020 at the glaucoma unit of the affiliated Changshu Hospital of Xuzhou Medical University. All patients were followed up for at least 12 months postoperatively. The preoperative to postoperative changes in best-corrected visual acuity (BCVA), intraocular pressure (IOP), number of antiglaucoma medications, and surgery-associated complications were recorded. The BCVA improved from 1.15 ± 0.91 logMAR preoperatively to 0.86 ± 0.82 logMAR at the final postoperative examination (Z = -3.62, P < 0.0001). The mean IOP was 36.63 ± 13.50 mmHg preoperatively and 15.14 ± 3.19 mmHg at the final examination (Z = -5.16, P < 0.0001). The number of antiglaucoma drugs was significantly reduced from 2.23 ± 0.55 preoperatively to 0.54 ± 0.86 at the final postoperative examination (Z = -5.26, P < 0.0001). The absolute value of the mean defect and retinal nerve fiber layer thickness at the last follow-up postoperatively were significantly reduced compared to preoperative values (Z = -3.35, P=0.001; Z = -4.56, P < 0.001, respectively). One patient experienced an explosive suprachoroidal hemorrhage during the operation. The sclera was incised at the corresponding site of the intraoperative hemorrhage. The operation was continued once there was no active bleeding, and the outcome was satisfactory. None of the patients required additional surgery to treat complications. Thus, cyclophotocoagulation under microscopic direct vision combined with phacoemulsification can be performed safely for the management of primary angle-closure glaucoma.

Vitreous Decompression Combined with Phacoemulsification for Medically Unresponsive Acute Angle Closure.

The management of acute angle closure combined with an extremely shallow anterior chamber and cataract remains complex. This study evaluated a technique of vitreous needle aspiration combined with phacoemulsification for the treatment of acute angle closure with continuous high intraocular pressure (IOP). We retrospectively reviewed the results of vitreous needle aspiration combined with phacoemulsification in 17 eyes (17 patients) with acute angle closure with continuous high IOP and coexisting visually significant cataracts between September 2018 and April 2020 at the glaucoma unit of the affiliated Changshu Hospital of Xuzhou Medical University. The main outcomes were the best corrected visual acuity (BCVA), IOP, anterior chamber depth (ACD), angle open distance 500 (AOD500), number of antiglaucoma medications, and surgery-associated complications. There were no complications during phacoemulsification and a foldable acrylic intraocular lens was implanted in the capsular bag in all 17 patients. For all patients, vitreous needle aspiration was successful at the first attempt. The BCVA improved from 2.02 ± 0.54 logMAR preoperatively to 0.73 ± 0.57 logMAR postoperatively at the final examination (p < 0.001). The mean IOP was 54.47 ± 5.33 mmHg preoperatively and 15.59 ± 2.35 mmHg at the final examination (p < 0.001) without any medication. The ACD was 1.70 ± 0.16 mm preoperatively and 3.35 ± 1.51 mm at the final examination (p < 0.001). The AOD500 was 0.07 ± 0.02 mm preoperatively and 0.51 ± 0.04 mm at the final examination (p < 0.001). Our vitreous needle aspiration technique can be performed safely in phacoemulsification for the management of acute angle closure with continuous high IOP.

Cilostazol Promotes Angiogenesis and Increases Cell Proliferation After Myocardial Ischemia-Reperfusion Injury Through a cAMP-Dependent Mechanism.

PURPOSE: Previous study indicated the protective role of cilostazol in ischemia-reperfusion (I/R) injury. Here, we aimed to explore the function of cilostazol in myocardial I/R injury and the underlying mechanism. METHODS: The myocardial I/R injury rat model was constructed, and the expression levels of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor receptor b (PDGF-B) and the number of new blood vessels were measured by qRT-PCR and immunohistochemistry. VSMC and HUVEC cells were treated with hypoxia to induce in vivo I/R injury model. The protein expression of AKT, endothelial nitric oxide synthase (eNOS) and apoptosis-related protein levels were detected by western blotting. Besides, the positive staining rate and cell viability were tested by 5-bromo-2-deoxyuridine (Brdu)/4',6-diamidino-2-phenylindole (DAPI) or DAPI/TdT-mediated dUTP Nick-End Labeling (TUNEL) staining and MTT assay. RESULTS: Cilostazol promoted angiogenesis by increasing the number of new blood vessels and up-regulating the expression of VEGF, HGF, bFGF and PDGF-B in myocardial I/R-injury rat model. The in vitro experiments showed that cilostazol increased the level of eNOS and AKT, and also enhanced cell proliferation in hypoxia-treated VSMC and HUVEC cells. Furthermore, after 8-Br-cAMP treatment, VEGF, HGF, bFGF, PDGF-B, p-AKT and p-eNOS expression were up-regulated, while cleaved-caspase 3 and cleaved-PARP expression were down-regulated. In addition, the effects of cilostazol on cell viability and apoptosis were aggravated by 8-Br-cAMP and attenuated after KT-5720 treatment. CONCLUSION: Cilostazol could promote angiogenesis, increase cell viability and inhibit cell apoptosis, consequently protecting myocardial tissues against I/R-injury through activating cAMP.

Cilostazol protects against myocardial ischemia and reperfusion injury by activating transcription factor EB (TFEB).

Although cilostazol was proved to have antitumor biological effects, its function in myocardial ischemia and reperfusion (I/R) injury and the underlying mechanisms were not fully illustrated yet. In this study, a rat model of I/R injury was constructed and quantitative real-time PCR, Western blot, and immunofluorescence (IF) assay were performed. Our results showed that cilostazol increased LC3 II/LC3 I ratio, reduced p62 abundance, and promoted the expressions of LAMP1, LAMP2, cathepsin B, and cathepsin D, indicating that cilostazol could activate autophagy and elevated lysosome activation. Following analysis showed that cilostazol enhanced nuclear protein expression of transcription factor EB (TFEB), an important regulator of autophagy-lysosome pathway. Furthermore, CCI-779, an inhibitor of TFEB, could reverse the effects of cilostazol on autophagic activity and lysosome activation. Importantly, cilostazol suppressed I/R injury-induced apoptosis by decreasing the cleavage of caspase 3 and PARP. Enzyme-linked immunosorbent assay showed that cilostazol reduced the serum levels of CTn1 and CK-MB and decreased infract size caused by I/R injuries. Altogether this study suggested that cilostazol protects against I/R injury by regulating autophagy, lysosome, and apoptosis in a rat model of I/R injury. The protective mechanism of cilostazol was partially through increasing the transcriptional activity of TFEB.

Immunomodulatory Effects of Enzymatic-Synthesized α-Galactooligosaccharides and Evaluation of the Structure-Activity Relationship.

In this study, α-galactooligosaccharides (α-GOSs) were synthesized using galactose as the substrate and α-galactosidase from Aspergillus niger as the catalyst. In the reaction, synthesized products of U1, U2, U3, and U4 were detected by high-performance liquid chromatography. By mass spectrometry, nuclear magnetic resonance, and 1-phenyl-3-methyl-5-pyrazolone derivatization, U1 was the mixture of disaccharides of α-d-Gal p-(1→1)-α-d-Gal, α-d-Gal p-(1→2)-α-d-Gal, α-d-Gal p-(1→3)-α-d-Gal, α-d-Gal p-(1→4)-α-d-Gal, U2 was identified to be α-d-Gal p-(1→6)-α-d-Gal, U3 was the mixture of galactotrisaccharides linked by one α-(1→6)-glycosidic linkage and one other α-glycosidic linkage, and U4 was identified as α-d-Gal p-(1→6)-α-d-Gal p-(1→6)-α-d-Gal. Afterward, the synthesized α-GOSs (U1, U2, U3, U4, and their mixture) as well as α-GOSs (manninotriose, stachyose, ciceritol, and verbascose) obtained from natural materials were used as subjects to evaluate their immunomodulatory effects in vitro by culturing mouse macrophage RAW264.7 cells. The results showed that α-GOS with a higher degree of polymerization had better immunomodulatory activity, while to a certain extent, α-GOS linked with α-(1→6)-galactosidic linkage showed a better immunomodulatory effect.

Engineering sulfur storage in maize seed proteins without apparent yield loss.

Sulfur assimilation may limit the pool of methionine and cysteine available for incorporation into zeins, the major seed storage proteins in maize. This hypothesis was tested by producing transgenic maize with deregulated sulfate reduction capacity achieved through leaf-specific expression of the Escherichia coli enzyme 3'-phosphoadenosine-5'-phosphosulfate reductase (EcPAPR) that resulted in higher methionine accumulation in seeds. The transgenic kernels have higher expression of the methionine-rich 10-kDa δ-zein and total protein sulfur without reduction of other zeins. This overall increase in the expression of the S-rich zeins describes a facet of regulation of these proteins under enhanced sulfur assimilation. Transgenic line PE5 accumulates 57.6% more kernel methionine than the high-methionine inbred line B101. In feeding trials with chicks, PE5 maize promotes significant weight gain compared with nontransgenic kernels. Therefore, increased source strength can improve the nutritional value of maize without apparent yield loss and may significantly reduce the cost of feed supplementation.

Cilostazol protects mice against myocardium ischemic/reperfusion injury by activating a PPARγ/JAK2/STAT3 pathway.

Myocardial ischemia/reperfusion (MIR) injury causes severe arrhythmias and a high lethality. The present study is designed to investigate the effect of cilostazol on MIR injury and the underlying mechaism. We measured the effects of cilostazol on heart function parameters in a mouse model of MIR. Proinflammatory cytokines and apoptosis proteins in the myocardium were examined to investigate the anti-inflammatory and anti-apoptosis ability of cilostazol. The participation of PPARγ/JAK2/STAT3 pathway was investigated. Results showed that the impairment of hemodynamic parameters caused by MIR was attenuated by cilostazol. The IL-6, IL-1ß and TNF-a levels were all decreased by cilostazol. Cilostazol also significantly inhibited Bax and cleaved caspase-3 levels and restored the Bcl-2 levels. PPARγ, JAK2 and STAT3 were all activated by cilostazol. Treatment of inhibitors of them abolished the protective effects of cilostazol on cardiac function, myocardial inflammation and apoptosis. In summary, cilostazol alleviated the cardiac function impairment, myocardial inflammation and apoptosis induced by MIR. The results present a novel signaling mechanism that cilostazol protects MIR injury by activating a PPARγ/JAK2/STAT3 pathway.

Somatic copy number losses on chromosome 9q21.33q22.33 encompassing the PTCH1 loci associated with cardiac fibroma.

Cardiac fibroma is an extremely rare benign tumor that remains poorly characterized genetically. Somatic copy number alterations are common in tumors and have been defined as a crucial factor leading to tumors. In this study, we present a child diagnosed with cardiac fibroma with somatic copy number losses of a total of three discontinuous segments from 9q21.33 to 9q22.33, including a mosaic deletion of PTCH1. PTCH1 has been associated with sporadic cardiac fibroma. Sequencing analysis of the PTCH1 gene has not revealed any causative mutation. Quantitative PCR analysis of PTCH1 further confirms somatic copy number losses. Our data narrow down the critical causative deletions for sporadic cardiac fibroma to a region more precise than any other previously reported one. Our results suggest important roles of somatic copy number losses on chromosome 9q21.33q22.33 in the development of sporadic cardiac fibroma; these findings may provide a better understanding of sporadic cardiac fibroma pathogenesis and contribute to the identification of novel diagnostic biomarkers of this neoplasm. .

A luciferase-based method for assay of 5'-adenylylsulfate reductase.

A luciferase-based method was developed for measurement of 5'-adenylylsulfate (APS) reductase (APR), an enzyme of the reductive sulfate assimilation pathway in prokaryotes and plants. APR catalyzes the two-electron reduction of APS and forms sulfite and adenosine 5'-monophospahate (AMP). The luciferase-based assay measures AMP production using an enzyme-coupled system that generates luminescence. The method is shown to provide an accurate measurement of APR kinetic properties and can be used for both endpoint and continuous assays. APR activity can be measured from pure enzyme preparations as well as from crude protein extracts of tissues. In addition, the assay is ideally suited to high-throughput sample analysis of APR activity in a microtiter dish format. The method adds new capability to the study of the biochemistry and physiology of APR.