Protective effects of curcumin on corneal endothelial cell PANoptosis and monocyte adhesion induced by tumor necrosis factor-alpha and interferon-gamma in rats.

Decrease of human corneal endothelial cell (CEC) density leads to corneal edema, progressive corneal opacity, and reduced visual acuity. A reduction in CEC density may be related to elevated levels of inflammatory cytokines, such as tumor necrosis factor (TNF)-α and interferon (INF)-γ. PANoptosis, characterized by the activation of apoptosis, necroptosis, and pyroptosis, could be a factor in the loss of CECs driven by TNF-α and INF-γ. Cytokines also stimulate monocytes adhesion to endothelium. It has been shown in previous research that curcumin plays protective roles against numerous corneal inflammatory diseases. However, it is not determined whether curcumin acts as an anti-PANoptotic agent or if it mitigates monocyte adhesion to CECs. Therefore, this research aimed to explor the potential therapeutic effects of curcumin and its underlying mechanisms in the loss of CECs. CEC injury models were established, and curcumin was injected subconjunctivally. Clinical evaluation of the corneas was conducted using a scoring system and anterior segment photography. Corneal observation was performed with hematoxylin and eosin staining and immunostaining of zona occludens-1(ZO-1). Apoptotic cells within the corneal endothelium were observed using TUNEL staining. The detection of primary proteins expression was accomplished through Western blot analysis. Interleukin (IL)-1ß and macrophage chemotactic protein 1 (MCP-1) levels were determined via ELISA, while the expression of cleaved caspase-3, gasdermin-D (GSDMD), phosphor-mixed lineage kinase domain-like protein (p-MLKL) and intercellular cell adhesion molecule-1 were confirmed by immunofluorescence. Myeloperoxidase (MPO) activity was measured in aqueous humors. Curcumin treatment attenuated the loss of CECs and corneal edema caused by TNF-α and IFN-γ. Besides, it decreased the count of TUNEL-positive cells, and inhibited the upregulation of cleaved caspase-3, cleaved caspase-6, cleaved caspase-7, and cleaved poly (ADP-ribose) polymerase. Moreover, both the expression and phosphorylation of MLKL and receptor-interacting protein 3 were decreased in curcumin-treated rats. Furthermore, curcumin also lowered the expression of cleaved caspase-1, diminished the levels of IL1ß and MCP-1, and inhibited the activity of MPO. Besides, the expression of intercellular cell adhesion molecule-1, vascular cell adhesion molecule-1, as well as the number of CD11b-positive cells adhered to the CECs decreased for the administration of curcumin.

Resveratrol regulates macrophage recruitment and M1 macrophage polarization and prevents corneal allograft rejection in rats.

Introduction: Resveratrol is an immune modulator that can reduce M1 macrophage polarization in vitro. Reducing macrophage recruitment and M1 polarization can prevent corneal allograft rejection (CGR). In this study, rat corneal allograft rejection models were established to explore the effects of resveratrol on CGR and macrophages and the underlying mechanisms after corneal transplantation. Methods: Corneal allograft models were established, and 100 mg/kg resveratrol was injected intraperitoneally. The corneal allografts were assessed clinically using the Holland rejection scoring system, anterior segment photography, and anterior segment optical coherence tomography. Corneal macrophages, pro-inflammatory cytokines, and corneal lymphatic vessels were detected using hematoxylin and eosin staining, immunofluorescence staining, and real-time quantitative polymerase chain reaction (qRT-PCR). Dendritic cells (DCs) in cervical lymph nodes were explored using flow cytometry. RNA sequencing experiments were conducted to identify the mechanisms through which resveratrol affected CGR. The results were verified using Simple Western analysis. Pro-inflammatory cytokines by macrophages in vitro were measured using qRT-PCR and enzyme-linked immunosorbent assays. Results: Resveratrol significantly prolonged the survival of corneal grafts and reduced graft edema and central corneal thickness. Corneal macrophage recruitment and M1 macrophage polarization decreased significantly after corneal transplantation in the resveratrol group. Resveratrol also reduced pro-inflammatory cytokines in corneal grafts and suppressed the early generation of cornea lymphatic vessels and the recruitment of cornea inflammatory cells 14 days after surgery. Resveratrol decreased the proportion of DCs in ipsilateral cervical lymph nodes. The effect of resveratrol on CGR was related to the phosphatidylinositol 3-kinase/protein kinase-B (PI3K/Akt) pathway. Resveratrol reduced the secretion of pro-inflammatory cytokines by M1 macrophages in vitro. Conclusion: Our findings suggest that resveratrol can reduce corneal macrophage recruitment and M1 macrophage polarization after corneal transplantation in rats and prevent CGR. The PI3K/Akt pathway may be an important mechanism that warrants further research.

Botulinum toxin treatment for bielschowsky acquired commitant esotropia in adults.

BACKGROUND: Many researchers have noticed that there is an increasing trend of Bielschowsky acquired comitant esotropia (ACE) in recent years related to excessive near work, but the exact pathogenesis and treatment methods have not been reported yet. Therefore, we aimed to characterize the clinical features of this ACE in adults and to evaluate the efficacy of botulinum toxin (BTX) injections in these patients. METHODS: This was a prospective consecutive case series of 47 patients with Bielschowsky ACE. BTX was injected bilaterally into the medial rectus muscle of 45 patients, and twenty-seven of them (27/45) completed 10 months of follow-up after their last injection. Angle of deviation, fusion, stereopsis, subjective assessment of diplopia were documented before and after BTX treatment, and repeated measures data were compared by the Wilcoxon signed-rank test or Analysis of variance. The relationship between BTX dosage and corrected esotropia was explored by the Regression analysis. Meanwhile, possible risk factors for ACE including time spent on near work, refraction error, patients' personality, glasses wearing habits and duration of symptoms were recorded and analyzed with General Linear Models. RESULTS: The patients aged 32.32 ± 10.96 (range 15-53) years spent 8.34 ± 2.38 h on near work each day, and most myope habitually removed their glasses at near. Their chief complaint was distance diplopia, with more significant esotropia at distance (around 20 PD) than at near. This series of patients also exhibited perfectionist tendencies. However, most patients achieved orthophoria after BTX treatment, only with a mild residual esotropia (+ 3.96 ± 5.79 PD), which left them asymptomatic most of the time. CONCLUSION: This group of ACE patients was characterized by diplopia with more significant esotropia at distance. Besides excessive near-work, habitually removing myopic glasses and perfectionist tendencies may also contribute to this type of ACE. Fortunately, bilateral BTX injection safely and effectively reduced the esotropia with complete resolution of symptoms, especially for those treated at an early stage.

Clinical features and treatment of near-work-related acquired esotropia.

AIM: To investigate the characteristics of near-work-related esotropia and the clinical efficacy of botulinum toxin type A (BTXA) injection therapy on it. METHODS: A total of 107 patients aged 15-57y with near-work-related esotropia were taken as the BTXA-treated group, and 30 other peers without near-work-related esotropia were included in the control group. All participants were refractive corrected to analyse the clinical characteristics of near-work-related esotropia. All subjects were examined including Worth4 spot examination, stereoscopic vision, strabismus angle, accommodative convergence to accommodation ratio (AC/A), far and near positive and negative convergence, positive and negative fusion range, positive and negative relative accommodation. Clinical efficacy was evaluated at a period of 10mo follow-up. RESULTS: The distant and near stereopsis were found in 84.9% and 77.5% of patients in the BTXA-treated group, respectively. In the control group, all patients had distant and near stereopsis. The incidence of taking off one's glasses to see close objects was significantly higher in the BTXA-treated group than in the control group (P<0.05). The BTXA-treated group showed a smaller range of in-fusion (9.84±5.72)° than the control group (22.04±8.71)° (P<0.05). The near esotropia angle of the BTXA-treated group (17.08±11.98)Δ was significantly smaller than the distant esotropia angle (19.07±11.68)Δ (P<0.05). Ten months after injection, the diplopia and esotropia of most patients underwent improvements after treatment (P<0.05). CONCLUSION: This study reveals that the accommodation function and the habit of near work without wearing spectacles are associated with near-work-related esotropia, while the length of time for near work and the onset time are independent of near-work-related esotropia. Additionally, BTXA injection therapy plays a vital role in relieving diplopia and restoring eye position.

Treatment of Pelvic Organ Prolapse by the Downregulation of the Expression of Mitofusin 2 in Uterosacral Ligament Tissue via Mesenchymal Stem Cells.

Background: The relationship between pelvic organ prolapse (POP), an aging-related disease, and the senescence-related protein mitofusin 2 (Mfn2) has rarely been studied. The aim of the present study was to explore the therapeutic effects of the downregulation of Mfn2 expression by stem cells on POP through animal experiments. Methods: First, a rat POP model was constructed by ovariectomy and traction. The rats in the non-pelvic organ prolapse (NPOP) and POP groups were divided into four groups for negative controls (N1−N4, N1: NPOP-normal saline; N2: NPOP-untransfected stem cells; N3: NPOP-short hairpin negative control (NPOP-sh-NC); N4: NPOP-short hairpin-Mfn2 (NPOP-sh-Mfn2)), and four groups for prolapse (P1−P4, P1: POP-normal saline; P2: POP-untransfected stem cells; P3: POP-sh-NC; P4: POP-sh-Mfn2), respectively. Stem cells were then cultured and isolated. The expression of Mfn2 was inhibited by lentivirus transfection, and the stem cells were injected into the uterosacral ligament of the rats in each group. The expression levels of Mfn2 and procollagen 1A1/1A2/3A1 in the uterosacral ligaments of the rats were observed at 0, 7, 14, and 21 days after injection. Results: Compared to the rats in the NPOP group, the POP rats had significant prolapse. The Mfn2 expression in the uterosacral ligaments of the POP rats was significantly increased (p < 0.05, all), and the expression of procollagen 1A1/1A2/3A1 was significantly decreased (p < 0.001, all). The POP rat model maintained the same trend after 21 days (without stem cell injection). At day 14, compared to the rats in the N1 group, the Mfn2 expression in the uterosacral ligament of the rats in the N4 group was significantly decreased (p < 0.05, all), and the expression of procollagens was significantly increased (p < 0.05, all). Similarly, compared to the rats in the P1 group, the Mfn2 expression in the uterosacral ligament of the rats in the P4 group was significantly decreased (p < 0.05, all), and the expression of procollagens was significantly increased (p < 0.05, all). Similarly, on day 21, the Mfn2 mRNA and protein expression in the uterosacral ligament of the POP and NPOP rats was significantly decreased (p < 0.05, all), and the expression of procollagens was significantly increased (p < 0.05, all) in the rats in the sh-Mfn2 group (N4, P4) compared to the rats in the saline group (N1, P1). Conclusions: The downregulation of Mfn2 expression by stem cells decreased the expression of Mfn2 and increased the expression of procollagen1A1/1A2/3A1 in the uterosacral ligament of the POP rats; this effect was significant 14−21 days after the injection. Thus, Mfn2 may be a new target for POP control.

Identification of bioactive natural products using yeast：Application to monoacylglycerol lipase inhibitor extraction from Corydalis Rhizoma.

Identification of bioactive principles from natural products is considered a challenging task in drug discovery. Recently, ligand fishing has been growing in interest as a sustainable strategy. In this study, a yeast-based drug discovery strategy was investigated to screen and fish active ingredients from natural products. Human monoacylglycerol lipase (MAGL) was first displayed on the cell wall of Pichia pastoris. The expression of MAGL on the cell surface was confirmed by immunofluorescence analysis. The activity toward 7-HCA which was consistent with free enzymes in solution. Recombinant yeast strains were used to screen the potential inhibitors from traditional Chinese medicines. Preliminary screening showed that the extracts of 12 herbs showed inhibition on MAGL activity, among which Corydalis Rhizoma had the highest inhibition rate of 60.66 ± 2.44%. Recombinant yeast fishing combined with HPLC-Q-TOF-MS/MS analysis was further used to identify the potential MAGL inhibitors. Finally, dehydrocorydaline (DHC) in Corydalis Rhizoma was determined as a ligand to MAGL with the IC50 value at 154.7 µΜ. Corydalis Rhizoma has been used as a pain reliever in TCM. Intraperitoneal injection of 7 mg kg- 1 DHC in chronic constriction injury model rats significantly attenuated the mechanical allodynia and thermal hyperalgesia. Meanwhile, 2-arachidonoylglycerol, the major MAGL substrate in the brain, was significantly increased both in the hippocampus and striatum. In conclusion, yeast-based ligand fishing combined with HPLC-Q-TOF-MS/MS is a powerful strategy for drug discovery in complex mixtures and DHC from Corydalis Rhizoma was confirmed with high inhibitory activity to MAGL either in vitro or in vivo .

Synthesis of a Novel Flame Retardant Containing Phosphorus, Nitrogen, and Silicon and Its Application in Epoxy Resin.

A novel flame retardant (TDA) containing phosphorus, nitrogen, and silicon was synthesized successfully via a controllable ring-opening addition reaction between 1,3,5-triglycidyl isocyanurate, 9,10-dihydro-9-oxa-10-phosphaphenanthrene-10-oxide, and 3-aminopropyltriethoxysilane, and TDA was then blended with diglycidyl ether of bisphenol A to prepare flame-retardant epoxy resins (EPs). The chemical structure and components of TDA were confirmed by Fourier transform infrared (FTIR) spectra, 31P nuclear magnetic resonance, and X-ray photoelectron spectroscopy. Thermogravimetric analysis results indicated that after the introduction of TDA, cured EP maintained good thermal stability with a minimum initial decomposition temperature of 337.6 °C, and the char yields of a EP/TDA-5 sample significantly increased by 76.2% compared with that of the neat EP thermoset. Additionally, with the addition of 25.0 wt % TDA (1.05 wt % phosphorus loading), the limited oxygen index value of cured EP increased from 22.5% of pure EP to 33.4%, and vertical burning V-0 rating was easily achieved. Meanwhile, after the incorporation of TDA, the total heat release and total smoke production of the EP/TDA-5 sample obviously reduced by 28.9 and 27.7% in the cone calorimeter test, respectively. Flame-retardant performances and flame-retardant mechanisms were further analyzed by scanning electron microscopy, FTIR, energy-dispersive spectrometry, and pyrolysis gas chromatography/mass spectrometry. The results reveal that the synergistic effect of phosphorus, nitrogen, and silicon plays an excellent flame-retardant role in both gaseous and condensed phases. In addition, the mechanical and dynamic mechanical properties of cured EP thermosets are well maintained rather than destroyed. All the results demonstrate that TDA endows epoxy resin with excellent flame retardancy and possesses great promise in the industrial field.

[Research Advance on the Factors Influencing Klebsiella Pneumoniae Biofilm Formation and the Regulation Mechanisms].

The biofilm formation is a dynamic process of bacterial growth,and the extracellular components can encase these microorganisms,making them more resistant to antibiotics and host immune attack.The formation of antibiotic-resistant bacterial biofilms will be a major challenge for the treatment and control of clinical infections.Klebsiella pneumoniae is a common clinical pathogen.Hypervirulent K.pneumoniae and carbapenem-resistant K.pneumoniae can cause serious infectious diseases,and the formation of its biofilm will aggravate this situation.In-depth research on the formation of K.pneumoniae biofilm and its regulatory mechanism can shed light on clinical anti-infection treatment and control.This article mainly reviews the latest research progress in the effects of fimbriae,polysaccharides,quorum sensing systems,and efflux pumps on the formation of K.pneumoniae biofilm and their regulatory mechanisms.

SIRT3 Transfection of Aged Human Bone Marrow-Derived Mesenchymal Stem Cells Improves Cell Therapy-Mediated Myocardial Repair.

Sirtuin 3 (SIRT3) is a deacetylase important for antioxidant protection, cell longevity, and aging. We hypothesized that SIRT3 improve oxidative resistance of aged cells and improve cell therapy in aged patients. In vitro, the proliferation and oxidative resistance of human mesenchymal stem cells (hMSCs) significantly declined with age. The expression and activity of antioxidant enzymes, including catalase (CAT) and manganese superoxide dismutase (MnSOD), increased after transfection of SIRT3 in hMSCs from older donors (O-hMSCs). The protein level of Forkhead box O3a (FOXO3a) in nucleus increased after SIRT3 overexpression. The antioxidant capacity of O-hMSCs increased after SIRT3 overexpression. 3-Amino-1,2,4-triazole (3-AT, CAT inhibitor) or diethyldithiocarbamate (DETC, SOD inhibitor) that was used to inhibit CAT or SOD activity significantly blocked the antioxidant function of SIRT3. When two inhibitors were used together, the antioxidant function of SIRT3 almost disappeared. Following myocardial infarction and intramyocardial injections of O-hMSCs in rats in vivo, the survival rate of O-hMSCs increased by SIRT3 transfection. The cardiac function of rats was improved after SIRT3-overexpressed O-hMSC transplantation. The infarct size, collagen content, and expression levels of matrix metalloproteinase 2 (MMP2) and MMP9 decreased. Besides, the protein level of vascular endothelial growth factor A and vascular density increased after cell transplantation with SIRT3-modified O-hMSCs. These results indicate that damage resistance of hMSCs decline with age and SIRT3 might protect O-hMSCs against oxidative damage by activating CAT and MnSOD through transferring FOXO3a into nucleus. Meanwhile, the therapeutic effect of aged hMSC transplantation can be improved by SIRT3 overexpression.

Contributions of Zn Ions to ZnO Nanoparticle Toxicity on Microcystis aeruginosa During Chronic Exposure.

In this work, we assessed the toxic effects of ZnO nanoparticles (NPs; 1, 10, and 50 mg L-1) and the corresponding dissoluble Zn ions (0.71, 8.66, and 35.59 mg L-1) on Microcystis aeruginosa. After chronic exposure (28 days), significantly higher growth inhibition was observed under ZnO NPs at 1 mg L-1 (47%) than under Zn ions at 0.71 mg L-1 (-15%). The opposite effect pattern was observed for ZnO NPs at 10 (71% vs. 80%) and 50 mg L-1 (73% vs. 95%) compared to Zn ions at the corresponding concentrations. After 7 days of exposure, ZnO NPs at 10 and 50 mg L-1 led to an increase of 83 and 53% in malondialdehyde content, as well as an increase of 106 and 61% in superoxide dismutase activity, respectively. However, Zn ions at the corresponding concentrations showed negligible impacts on the two parameters. The different results indicate that the insoluble NPs during the initial exposure mostly account for lipid peroxidation, which further lead to microalgal antioxidant response. During the subsequent exposure, the contributors of ZnO NP toxicity shift with the concentration and exposure time of ZnO NPs. In conclusion, the study presents new insights into the different contributions of insoluble NPs and dissoluble metallic ions to metallic NP toxicity.

Harmful effect of nanoparticles on the functions of freshwater ecosystems: Insight into nanoZnO-polluted stream.

ZnO nanoparticle toxicity on aquatic organisms has been extensively studied, but its concentration-and time-dependent effects on ecosystem functioning are remain uncertain. Here we assessed the harmful effects of nano-ZnO (10, 100, 1000 mg L-1) on the stream functioning by using a microcosm system simulating poplar leaf decomposition for 50 days. The 100 mg L-1 ZnO nanoparticles had significantly and stably inhibitory effect on the litter decomposition during the exposure period. The inhibition was not detected in the 10 mg L-1 treatment until 43 d. In contrast, the significant and continuous inhibition started to disappear from 43 d in the 1000 mg L-1 treatment. The varied consequences on litter decomposition might be directly affected by the different ZnO nanoparticle homogeneity of the different treatments. ZnO nanoparticles led to significant decreases in pH value of the decomposition environment, which had significant and positive relationships to the activities of dehydrogenase, glycine-aminopeptidase, N-acetylglucosaminidase, and acid phosphatase. Besides, 10 and 1000 mg L-1 ZnO nanoparticles led to lower fungal diversity, which was negatively related to the variability of decomposition. In conclusion, fungal decomposers showed different responses to the different concentrations of ZnO nanoparticle, and ultimately affected the stability of ecosystem functions.

Genome-wide identification, characterization, and expression analysis of superoxide dismutase (SOD) genes in foxtail millet (Setaria italica L.).

Superoxide dismutases (SODs) play important roles in plant growth, development, and response to abiotic stresses. Despite SOD gene families have been identified in various plant species, little is known in foxtail millet (Setaria italica L.). In this study, a systematic analysis of SOD gene family was performed in foxtail millet and the expression pattern of SOD genes in response to abiotic stressors was analyzed at the whole-genomic level. Eight SOD genes were identified in foxtail millet, including 4 Cu/ZnSODs, 3 FeSODs, and 1 MnSOD. These SiSODs are unevenly distributed across 5 of the 9 chromosomes. Phylogenetic analysis showed that SOD proteins could be divided into two major categories (Cu/ZnSODs and Fe-MnSODs), containing seven subgroups, from foxtail millet and other plant species. SOD genes have conserved motif and exon/intron composition in the same subgroup among Setaria italica, Setaria viridis, and Oryza sativa. Additionally, many cis-elements that respond to different stressors were distributed at different densities in the promoters of 8 SiSODs. The expression patterns of SiSODs in different tissues and different abiotic stressors indicated that the SiSODs may play important roles in reactive oxygen species scavenging, caused by various stressors in foxtail millet. This study provides a foundation for the further cloning and functional verification of the SOD gene family response to environmental stimuli in foxtail millet.

[Influence of moxibustion of baihui (GV 20) on hemodynamics of common carotid artery in healthy subjects].

OBJECTIVE: To observe the effect of moxibustion of Baihui (GV 20) on the hemodynamics of internal carotid artery in health subjects so as to study its underlying mechanism of moxibustion in the treatment cerebrovascular disorders. METHODS: Thirty healthy male volunteer students between 20 and 22 years in age were enrolled into this study. Qm (mean blood flow), Vm) (mean velocity of blood flow), Vmax (maximal velocity of blood flow), Vmin (minimal velocity of blood flow), Wv (pulse wave velocity), Zcv (characteristic impedance), Rv (peripheral resistance), DR (dynamic resistance), CP (critical pressure) and DP (difference of pressure) of the right and left common carotid arteries were measured before and after moxibustion of GV20 (5-10 min each time, once daily, 5 times altogether) by using CBA CV-300 Cerebrovascular Hemodynamics Detector. RESULTS: Following moxibustion of GV20, Qm, Vm and Vmin of both right and left common carotid arteries increased significantly (P < 0.01); while Rv and DR of the brain artery lowed evidently (P < 0.05); The rest indexes had no significant changes (P > 0.05). CONCLUSION: Moxibustion of Baihui (GV 20) can significantly raise the velocity of blood flow of the common carotid artery and low the resistance of blood vessels.