Key genes and regulatory networks for diabetic retinopathy based on hypoxia-related genes: a bioinformatics analysis.

AIM: To prevent neovascularization in diabetic retinopathy (DR) patients and partially control disease progression. METHODS: Hypoxia-related differentially expressed genes (DEGs) were identified from the GSE60436 and GSE102485 datasets, followed by gene ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Potential candidate drugs were screened using the CMap database. Subsequently, a protein-protein interaction (PPI) network was constructed to identify hypoxia-related hub genes. A nomogram was generated using the rms R package, and the correlation of hub genes was analyzed using the Hmisc R package. The clinical significance of hub genes was validated by comparing their expression levels between disease and normal groups and constructing receiver operating characteristic curve (ROC) curves. Finally, a hypoxia-related miRNA-transcription factor (TF)-Hub gene network was constructed using the NetworkAnalyst online tool. RESULTS: Totally 48 hypoxia-related DEGs and screened 10 potential candidate drugs with interaction relationships to upregulated hypoxia-related genes were identified, such as ruxolitinib, meprylcaine, and deferiprone. In addition, 8 hub genes were also identified: glycogen phosphorylase muscle associated (PYGM), glyceraldehyde-3-phosphate dehydrogenase spermatogenic (GAPDHS), enolase 3 (ENO3), aldolase fructose-bisphosphate C (ALDOC), phosphoglucomutase 2 (PGM2), enolase 2 (ENO2), phosphoglycerate mutase 2 (PGAM2), and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3). Based on hub gene predictions, the miRNA-TF-Hub gene network revealed complex interactions between 163 miRNAs, 77 TFs, and hub genes. The results of ROC showed that the except for GAPDHS, the area under curve (AUC) values of the other 7 hub genes were greater than 0.758, indicating their favorable diagnostic performance. CONCLUSION: PYGM, GAPDHS, ENO3, ALDOC, PGM2, ENO2, PGAM2, and PFKFB3 are hub genes in DR, and hypoxia-related hub genes exhibited favorable diagnostic performance.

Case Report: Torsade de Pointes Induced by the Third-Generation Epidermal Growth Factor Receptor-Tyrosine Kinase Inhibitor Osimertinib Combined With Litsea Cubeba.

Torsades de Pointes (TdP) occurred in a 68-year-old female with epidermal growth factor receptor (EGFR) mutant lung cancer administered osimertinib, the third-generation EGFR tyrosine kinase inhibitor (TKI). Electrocardiogram (ECG) recorded at Tdp showed QT prolongation (QTc = 515 ms), to which a Traditional Chinese Medicine (TCM) named "Litsea Cubeba" may have contributed. After discontinuation of osimertinib and Litsea Cubeba, magnesium supplementation, potassium supplementation, lidocaine infusion, and the pacemaker frequency adjustment, Tdp terminated. However, QT prolongation sustained at discharge (QTc = 528 ms), partly because of the emergency use of amiodarone. Osimertinib may prolong the QT interval leading to TdP, especially when multiple risk factors to lengthen QT interval are incidentally overlapped. Thus, regular monitoring of ECG and appropriate management of concomitant drugs are highly recommended.

Peripheral blood inflammation indices are effective predictors of anastomotic leakage in elective esophageal surgery.

BACKGROUND: This study investigated the predictive value of peripheral inflammatory indices, including neutrophil count, lymphocyte count, platelet count, neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR), in anastomotic leakage during elective esophageal surgery. METHODS: This retrospective study included all patients who underwent esophagectomy for esophageal squamous cell carcinoma from 2016 to 2020 in our institution. The peripheral blood inflammatory indices were obtained on preoperative days 1-7 (PRD 1-7), and postoperative days 1-3 (POD 1-3) and 4-7 (POD 4-7). Univariate, multivariate logistic, and receiver operating characteristic curve analyses were conducted to evaluate the diagnostic value of these peripheral blood inflammatory indices. RESULTS: A total of 198 patients were included in the study, and 25 (13%) patients experienced anastomotic leakage. Multivariate analyses identified diet, neutrophil count, and PLR on POD 1-3, and NLR on POD 4-7 as independent factors associated with anastomotic leakage. Using the receiver operating characteristic curve, the variable with the best area under curve was a neutrophil cutoff count of 4.1 [0.737; 95% CI: 0.639-0.835], with a sensitivity and specificity of 60.0% and 66.5%, respectively. This was followed by an NLR cutoff value of 9.5 on POD 4-7 (0.628; 95% CI: 0.505-0.752) and a cutoff PLR value of 220.1 on POD 1-3 (0.643; 95% CI: 0.536-0.750). Diet showed a poor result on the receiver operating characteristic curve analysis. CONCLUSIONS: Neutrophil count and PLR on POD 1-3 and NLR on POD 4-7 were shown to have predictive value for anastomotic leakage in elective esophageal surgery.

[Water utilization strategy of three soil and water conservation trees on Ordos Plateau, China]. / 鄂尔多斯高原3ä¸ªæ°´åœŸä¿æŒæ ‘ç§çš„æ°´åˆ†åˆ©ç”¨ç­–ç•¥.

To understand the adaptive strategies of three tree species used for soil and water conservation in the hilly area of eastern Ordos Plateau, Hippophae rhamnoides, Pinus tabuliformis and Armeniaca sibirica, we measured the δ18O value of xylem water of branch and potential water sources and the δ13C value of leaves, and analyzed the relative contribution of each potential water sources to different tree species by multiple linear mixing model. We further compared the seasonal dynamic and inter-specific variation of water source and water use efficiency (WUE). The results showed that H. rhamnoides, P. tabuliformis and A. sibirica mainly used soil water in 10 cm layer in May, accounting for 88.5%, 94.0% and 91.6% of their total water uptake, respectively. In July, H. rhamnoides mainly used soil water in 10-25 cm layer (44.6%) and rain water (35.4%), P. tabuli-formis mainly used rain water (93.7%), and A. sibirica mainly used soil water in 25-100 cm layer (55.9%) and rain water (36.8%). In September, H. rhamnoides mainly used soil water in 25 cm and 75-100 cm layers (88.9%), P. tabuliformis mainly used soil water in 10 cm and 50-75 cm layers (84.5%), and A. sibirica used that in 10-100 cm layer. WUE of H. rhamnoides in May was significantly higher than that in July and September. WUE of P. tabuliformis in July was significantly higher than that in September. WUE of H. rhamnoides was significantly higher than that of P. tabuliformis and A. sibirica in May and July. Three tree species use different depths of soil water or rainwater based on their availability in different seasons. H. rhamnoides and P. tabuliformis could improve their WUE to adapt to environmental change under drought, which might be more suitable for local semi-arid environment than A. sibirica.

[Lycium barbarum Polysaccharide Pretreatment Attenuates Cerebral Ischemic Reperfusion Injury by Inhibiting Apoptosis in Mice].

OBJECTIVE: To observe the protective effects of Lycium barbarum polysaccharide(LBP) on focal cerebral ischemic reperfusion injury in mice, and to explore its mechanism. METHODS: Male mice were randomly divided into six groups: sham-operated group, middle cerebral artery occlusion(MCAO) mice group, MCAO mice treating with 4 mg/kg Nimodipine group and MCAO mice treating with 10, 20 and 40 mg/kg LBP groups. The mice were preventively administrated with LBP by intragastric administration for seven days. After 2 h of cerebral ischemia and 24 h of reperfusion, neurological scores in each group mice were estimated. Morphological changes in ischemic brain neurons were performed for HE staining. The number of apoptotic neurons was detected by Tunel staining. The Caspase-3 protein activity was measured by spectrophotometry. BAX and BCL-2 protein expressions in ischemic brains were investigated by Western blot analysis. RESULTS: Compared to the vehicle group, neurological deficit scores were significantly reduced in LBP pretreatment group(P <0.01). LBP( 10,20 and 40 mg/kg) groups relieved neuronal morphological damage respectively and also obviously attenuated the neuronal apoptosis (P <0. 05). Caspase-3 protein activity and BAX protein expression were obviously decreased(P <0. 05, P <0. 01) and BCL-2 protein expression was markedly increased(P <0. 01) in LBP pretreatment groups. CONCLUSION: LBP can protect against focal cerebral ischemic reperfusion injury in mice,the mechanism may be related with attenuating the apoptosis in ischemic brains.

Pinocembrin improves cognition and protects the neurovascular unit in Alzheimer related deficits.

Amyloid-ß (Aß) peptides accumulate in the brain and initiate a cascade of pathologic events in Alzheimer's disease. The receptor for advanced glycation end products (RAGE) has been implicated to mediate Aß-induced perturbations in the neurovascular unit (NVU). We demonstrated that pinocembrin exhibits neuroprotection through inhibition of the Aß and/or RAGE pathway, but the therapeutic role and mechanism involved are not ascertained. Here, we report that a 3-month treatment with pinocembrin prevents the cognition decline in APP/PS1 transgenic mice without altering Aß burden and oxidative stress. Instead, pinocembrin is effective in conferring neurovascular protection through maintenance of neuropil ultrastructure, reduction of glial activation and levels of inflammatory mediators, preservation of microvascular function, improving the cholinergic system by conserving the ERK-CREB-BDNF pathway, and modulation of RAGE-mediated transduction. Furthermore, in an in vitro model, pinocembrin provides the NVU protection against fibrillar Aß1₋42, accompanied by regulation of neurovascular RAGE pathways. Our findings indicate that pinocembrin improves cognition, at least in part, attributable to the NVU protection, and highlights pinocembrin as a potential therapeutic strategy for the prevention and/or treatment of Alzheimer's disease.

Pinocembrin protects brain against ischemia/reperfusion injury by attenuating endoplasmic reticulum stress induced apoptosis.

Endoplasmic reticulum stress (ER stress) is known to play a vital role in mediating ischemic reperfusion damage in brain. Our previous studies showed that pinocembrin alleviated cerebral ischemic injury in ischemia/reperfusion and vascular dementia animal models, but whether attenuation of ER stress-induced apoptosis contributes to the mechanisms remains to be elucidated. In this study, an attempt was therefore made to investigate the modulation effect of pinocembrin on ischemia/reperfusion-induced ER stress in brain. Focal cerebral ischemia/reperfusion rats were induced by middle cerebral artery occlusion (MCAO) for 2h followed by 6h reperfusion. Pinocembrin was administered in different doses (1mg/kg, 3mg/kg, and 10mg/kg, respectively) at the same time of onset of reperfusion. Neurological function and brain infarction were evaluated. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method, and flow cytometer (FCM) were used to investigate cell apoptosis in penumbra cortex. DNA fragmentation assay was also performed using electrophoresis. The expression of ER stress proteins of GRP78, CHOP/GADD153, ATF4, eIF2α phosphorylation was detected by western blot, and caspase-12 was evaluated by immunohistochemical analysis. Our results demonstrate that pinocembrin-treatment (3mg/kg and 10mg/kg) significantly reduced neurological deficit scores, infarct volume, and neuron apoptosis in the ischemia/reperfusion rats. It can also significantly modulate the protein levels by increasing GRP78 (10mg/kg) and attenuating CHOP/GADD153 expression along with caspase-12 activation (3mg/kg and 10mg/kg). At the same time, eIF2α phosphorylation was restrained and the expression of ATF4 was reduced (3mg/kg and 10mg/kg). These results suggest that the attenuation of ER stress induced apoptosis may be involved in the mechanisms of pinocembrin.

Quercetin protects against the Aß(25-35)-induced amnesic injury: involvement of inactivation of rage-mediated pathway and conservation of the NVU.

Quercetin has demonstrated protective effects against Aß-induced toxicity on both neurons and endothelial cells. However, whether or not quercetin has an effect on the neurovascular coupling is unclear. In the present study, we aim to investigate the anti-amnesic effects of quercetin and to explore the underlying mechanisms. Aß(25-35) (10 nmol) was administrated to mice i.c.v. Quercetin was administrated orally for 8 days after injection. Learning and memory behaviors were evaluated by measuring spontaneous alternation in Morris Water Maze test and the step-through positive avoidance test. The regional cerebral blood flow was monitored before the Aß(25-35) injection and on seven consecutive days after injection. Mice were sacrificed and cerebral cortices were isolated on the last day. The effects of quercetin on the neurovascular unit (NVU) integrity, microvascular function and cholinergic neuronal changes, and the modification of signaling pathways were tested. Our results demonstrate that quercetin treatment for Aß(25-35)-induced amnesic mice improved the learning and memory capabilities and conferred robust neurovascular coupling protection, involving maintenance of the NVU integrity, reduction of neurovascular oxidation, modulation of microvascular function, improvement of cholinergic system, and regulation of neurovascular RAGE signaling pathway and ERK/CREB/BDNF pathway. In conclusion, in Aß(25-35)-induced amnesic mice, optimal doses of quercetin administration were beneficial. Quercetin protected the NVU likely through reduction of oxidative damage, inactivation of RAGE-mediated pathway and preservation of cholinergic neurons, offering an alternative medication for Alzheimer's disease.

[Effects of yiqi huoxue qufeng decoction on the diamine oxidase and immunoglobin E of patients with chronic urticaria].

OBJECTIVE: To observe effects of Yiqi Huoxue Qufeng Decoction (YHQD, with the actions of replenishing qi, activating blood, and dispelling wind) on diamine oxidase (DAO) and immunoglobulin E (IgE) of patients with chronic urticaria. METHODS: Eighty-five chronic urticaria patients from the clinics of dermatology, Shaanxi Hospital of Traditional Chinese Medicine were randomly assigned to the treatment group (50 cases) and the control group (35 cases). Besides, another 15 healthy volunteers were recruited as the healthy group. Patients in the treatment group took YHQD, one dose daily, once in the morning and once in the evening. Patients in the control group took Fuyang Granule (FYG), 6 g each time, three times daily. The therapeutic course for the two groups was 8 weeks. The effective rates of the two groups were observed after treatment and 2 months after quitting treatment. The levels of DAO and IgE were observed in the three groups before and after treatment. RESULTS: The post-treatment recovery rate (20 cases, 44.0%) and the effective rate 2 months after quitting treatment (62.0%) were higher in the treatment group than in the control group (7 cases, 20.0%; 31.4%) with statistical difference (P<0.05). The DAO level in the two treatment groups (6.9 +/- 1.8 in the treatment group and 6.5 +/- 1.8 in the control group) was obviously higher than that in the healthy group (1.1 +/- 0.4), showing statistical difference (P<0.05). The post-treatment DAO and IgE both decreased in the treatment group and the control group when compared with before treatment in the same group. Those were lower in the treatment group than in the control group with statistical difference (P<0.05). CONCLUSION: YHQD could improve the symptoms of chronic urticaria patients, ameliorate the intestines mucosa barrier function and the immunity.

Pinocembrin protects against ß-amyloid-induced toxicity in neurons through inhibiting receptor for advanced glycation end products (RAGE)-independent signaling pathways and regulating mitochondrion-mediated apoptosis.

BACKGROUND: It is known that amyloid-ß peptide (Aß) plays a pivotal role in the pathogenesis of Alzheimer's disease (AD). Interaction between Aß and the receptor for advanced glycation end products (RAGE) has been implicated in neuronal degeneration associated with this disease. Pinocembrin, a flavonoid abundant in propolis, has been reported to possess numerous biological activities beneficial to health. Our previous studies have demonstrated that pinocembrin has neuroprotective effects on ischemic and vascular dementia in animal models. It has been approved by the State Food and Drug Administration of China for clinical use in stroke patients. Against this background, we investigated the effects of pinocembrin on cognitive function and neuronal protection against Aß-induced toxicity and explored its potential mechanism. METHODS: Mice received an intracerebroventricular fusion of Aß25-35. Pinocembrin was administrated orally at 20 mg/kg/day and 40 mg/kg/day for 8 days. Behavioral performance, cerebral cortex neuropil ultrastructure, neuronal degeneration and RAGE expression were assessed. Further, a RAGE-overexpressing cell model and an AD cell model were used for investigating the mechanisms of pinocembrin. The mechanisms underlying the efficacy of pinocembrin were conducted on target action, mitochondrial function and potential signal transduction using fluorescence-based multiparametric technologies on a high-content analysis platform. RESULTS: Our results showed that oral administration of pinocembrin improved cognitive function, preserved the ultrastructural neuropil and decreased neurodegeneration of the cerebral cortex in Aß25-35-treated mice. Pinocembrin did not have a significant effect on inhibiting Aß1-42 production and scavenging intracellular reactive oxygen species (ROS). However, pinocembrin significantly inhibited the upregulation of RAGE transcripts and protein expression both in vivo and in vitro, and also markedly depressed the activation of p38 mitogen-activated protein kinase (MAPK)-MAPKAP kinase-2 (MK2)-heat shock protein 27 (HSP27) and stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK)-c-Jun pathways and the downstream nuclear factor κB (NFκB) inflammatory response subsequent to Aß-RAGE interaction. In addition, pinocembrin significantly alleviated mitochondrial dysfunction through improving mitochondrial membrane potential and inhibiting mitochondrial oxidative stress, and regulated mitochondrion-mediated apoptosis by restoration of B cell lymphoma 2 (Bcl-2) and cytochrome c and inactivation of caspase 3 and caspase 9. CONCLUSIONS: Pinocembrin was shown to infer cognitive improvement and neuronal protection in AD models. The mechanisms of action of the compound were illustrated on RAGE-dependent transduction inhibition and mitochondrion protection. It appears to be a promising candidate for the prevention and therapy of AD.

Targeting the neurovascular unit: development of a new model and consideration for novel strategy for Alzheimer's disease.

Alzheimer's disease involves the complex and interconnected cascade of cellular and molecular events. Only a few treatments are available to slow the course of the disease at present. Recent studies suggest that neurovascular unit serves to maintain cerebral homeostasis, and pathological interactions between components of neurovascular unit lead to cerebral dysfunction. In present study, we established a functional unit trying to target major components of the neurovascular unit by the co-culture of rat cortical parenchymal culture and cerebral microvascular endothelial cells. This entity allowed the application of techniques such as immunofluorescent imaging and biological assays under defined conditions. The morphology of cell types, blood-brain barrier function and neuronal activation were investigated. The insight revealed that targeting components of the neurovascular unit, rather than just the neuron, might be a priority in Alzheimer's disease and more likely to provide cerebroprotection.

[Effects of intranasal interferon gamma on transforming growth factor-ß1/Smad in rats with allergic rhinitis].

OBJECTIVE: To investigate the effects of intranasal interferon gamma (IFN-γ) on nasal mucosa remodeling and expression of transforming growth factor-ß1 (TGF-ß1), Smad2, Smad3, Smad7 in allergic rhinitis (AR) rat model. METHODS: Ovalbumin (OVA) and aluminum hydroxide were used to construct the AR model. Thirty AR rats were randomly divided into positive control group (group B, n = 10), IFN-γ treatment group (group C, n = 10) and negative control group (normal rats, n = 10). After the AR models were built, 50 µl PBS, 1 µg IFN-γ was dropped into the nasal cavity of each rat in group B and group C, from the fouth week to tenth week, twice a week. The nasal mucosa was collected on day 71 in order to observe the pathologic changes, and the expression of TGF-ß1, TGF-ß1 mRNA, Smad2 mRNA, Smad3 mRNA and Smad7 mRNA by immunohistochemistry and reverse transcriptase-polymerase chain reaction. RESULTS: Decreases of TGF-ß1, Smad2 and Smad3 mRNA were seen in nasal tissue of group C (0.59 ± 0.04, 0.39 ± 0.08, 0.46 ± 0.15) as compared with group B (0.82 ± 0.12, 0.70 ± 0.18, 0.95 ± 0.26), the differences were significant (q value were 3.15, 4.47, 3.03, all P < 0.05). The levels of Smad7 mRNA expression increased significantly (q = 2.98, P < 0.05) in group C (0.31 ± 0.05) as compared with group B (0.25 ± 0.06). Immunohistochemistry showed significant decrease of TGF-ß1 expression in the nasal tissue of group C much lesser than that in group B. CONCLUSIONS: Intranasal IFN-γ could decrease the expression of TGF-ß1, TGF-ß1 mRNA, Smad2 mRNA, Smad3 mRNA, increase the expression of Smad7 mRNA in AR rats model and inhibit the nasal mucosa remodeling.

[Protective effect of effective parts of Zingiber Offecinal on vascular endothelium of the experimental hyperlipidemic rats].

OBJECTIVE: To observe the influence of effective parts of Zingiber Officinale on serum IL-6, TNF-alpha in oroler to investigate the protective effects of the effective parts of Zingiber Officinal (EPZ) on endothelium of the experimental hyperlipidemic rats and the mechanism of its effects. METHODS: The hyperlipidemia model of rats was constructed by feeding high-fat forage and filled with the effective parts of Zingiber Officinale 200 mg/kg, 400 mg/kg, 800 mg/kg every day for 13 weeks. Blood was drawn to determine both the level of serum IL-6 and TNF-alpha. All the aortaes were taken to oberserve morphologic change and the intima-media thickness were detected. RESULTS: The effective parts of Zingiber Officinale could markedly decrease intima-media thickness, but had no marked influence in the level of serum IL-6 and TNF-alpha. CONCLUSION: The Effect Parts of Zingiber Officinale has the effect of protection of the endothelia of hyperlipidemia rats, which has nothing with the level of serum IL-6 and TNF-alpha.

Tropical application of halcinonide cream reduces the severity and incidence of intraperitoneal adhesions in a rat model.

BACKGROUND: Systemic or intraperitoneal administration of corticosteroids has been reported to have conflicting effects on the prevention of peritoneal adhesions. Painting corticosteroid cream directly on the likely site of adhesion formation, owing to its high concentrations and persistent effects, may be a promising approach to prevent peritoneal adhesion formation. METHODS: Adhesions were induced by abrading of the cecum, followed by dropping of 95% ethanol. Sixty Wistar rats were randomly allocated to two control groups with no further treatment of the cecum and to two therapy groups treated with 0.1% halcinonide cream painted directly on the damaged surface of the cecum. After 3 and 7 days, adhesion scores, adhesion incidence, and intraperitoneal leukocytes were evaluated. RESULTS: On both postoperative days 3 and 7, halcinonide cream resulted in a significant decrease in mean adhesion scores (6.80 versus 0.67, 10.40 versus 1.26; P <0.001, P <0.001, respectively). The adhesion incidence was 43.3% for the therapy groups and 100% for controls (P <0.01). On day 3, the total numbers of intraperitoneal leukocytes were 120.73 +/- 24.01 millions for the therapy groups and 270.40 +/- 34.68 for controls (P <0.001). CONCLUSIONS: Painting halcinonide cream directly on the damaged surface of the cecum could effectively reduce the severity and incidence of adhesion, possibly by suppression of early inflammatory exudate and of late fibroblast invasion and proliferation.