The effect of ultrasound-guided intercostal nerve block on postoperative analgesia in thoracoscopic surgery: a randomized, double-blinded, clinical trial.

BACKGROUND: Intercostal nerve block (ICNB) is a very effective analgesic method. We aimed to explore the effect of preemptive analgesia with ultrasound-guided intercostal nerve block on postoperative analgesia in thoracoscopic surgery. METHODS: 126 patients, aged 18-70 years, with American Society of Anesthesiologists (ASA) physical status I-II and scheduled for thoracoscopic pulmonary resection were enrolled in this study. 119 patients were left for final analysis. Patients were randomly allocated to group ICNB and group CONTROL. Patients in CONTROL group were administered sufentanil with patient-controlled analgesia device after operation In group ICNB, patients received ropivacaine ICNB prior to surgery and patient-controlled analgesia device after operation. The primary outcome is visual analog scale pain score (VAS) at rest at 0,4, 8,16,24,48,72 and 168 h postoperatively and they were compared. Surgical outcomes and rescue analgesia requirement were also recorded. RESULTS: VAS scores were statistically significantly lower for ICNB group compared to control group at 0, 4, 8, 16, 24 and 48 h postoperatively. The duration of insertion of chest tube in ICBN group was shorter than that in control group, and the difference was statistically significant (4.69 ± 2.14 vs. 5.67 ± 2.86, P = 0.036). The postoperative hospital stay, incidence of nausea and vomiting and postoperative pulmonary infection rate in ICBN group were all lower than those in the control group, but there were no statistical differences. The frequency of rescue analgesia during 48 postoperative hours was different between the two groups (ICNB vs. Control; 9.83% vs. 31.03%, P = 0.004). CONCLUSIONS: For patients undergoing thoracoscopic surgery, ultrasound-guided ICNB is simple, safe, and effective for providing acute postoperative pain management during the early postoperative stage. TRIAL REGISTRATION: Chinese clinical trials: chictr.org.cn, ChiCTR1900021017. Registred on 25/01/2019.

Steroid Drugs as an Adjunct for Reducing the Incidence of Proliferative Vitreoretinopathy after Rhegmatogenous Retinal Detachment Surgery: A Meta-Analysis of Randomized Controlled Studies.

INTRODUCTION: The efficacy and influence of steroids for reducing the incidence of proliferative vitreoretinopathy (PVR) after rhegmatogenous retinal detachment (RRD) surgery remain controversial. Systematic review and meta-analysis were conducted to explore the effect of steroids versus placebo on risk of PVR. METHODS: We searched PubMed, Embase, Web of science, EBSCO, and Cochrane library databases through September 2020 for randomized controlled trials (RCTs), assessing the effect of steroid drugs as an adjunct for reducing the incidence of PVR after RRD surgery. This meta-analysis was performed using the random-effect model. Data were extracted by two reviewers independently; the quality of RCTs was assessed by the Cochrane risk-of-bias tool. We calculated risk ratio (RR) and the 95% confidence intervals (CIs) of all outcomes and plotted on forest plots. I2 accessed using the χ2 test was applied to quantify the degree of heterogeneity. RESULTS: Four RCTs involving 478 patients (478 eyes) are included in the meta-analysis. There was no significant difference in the incidence of PVR recurrence between steroid groups and control groups (RR: 0.87, 95% CI: 0.70-1.08, p = 0.19). However, the incidence of recurrent PVR was lower in the steroid group (RR: 0.67, 95% CI: 0.46-0.99, p = 0.04) than in the control group when only PVR grades A and B were taken into consideration. Besides, steroids could significantly reduce the incidence of macular edema after surgery (RR: 0.64, 95% CI: 0.47-0.88, p = 0.007). The steroid group and control group had comparable outcomes of retinal reattachment rate and reoperation rate after primary surgery. Additionally, there was no significant difference of the incidence of epiretinal membrane, and the incidence of surgery required by epiretinal membrane. CONCLUSION: This meta-analysis reveals that RRD surgery combined with steroid drugs administration could significantly reduce the recurrence in PVR grade A and B subgroup, as well as the incidence of macular edema after surgery.

Proteomic profiling and correlations with clinical features reveal biomarkers indicative of diabetic retinopathy with diabetic kidney disease.

Diabetic retinopathy (DR) and diabetic kidney disease (DKD) are complications of diabetes and place serious health and economic burdens on society. However, the identification and characterization of early biomarkers for DKD, especially for nonproliferative DR (NPDR) patients with DKD, are still needed. This study aimed to demonstrate the plasma proteomic profiles of NPDR+DKD and NPDR patients and identify potential biomarkers for early diagnosis of DKD. Fifteen plasma samples from the NPDR group and nine from the NPDR+DKD group were analyzed by LC-MS/MS to identify the differentially expressed proteins between the two groups. Functional enrichment, protein-protein interaction and clinical feature correlation analyses revealed the target protein candidates, which were verified using ELISA and receiver operating characteristic (ROC) analysis. In total, 410 proteins were detected in plasma; 15 were significantly upregulated and 7 were downregulated in the NPDR+DKD group. Bioinformatics analysis suggested that DKD is closely related to cell adhesion and immunity pathways. ß-2-Microglobulin (B2M) and vimentin (VIM) were upregulated in NPDR+DKD, enriched as hub proteins and strongly correlated with clinical features. ELISA showed that B2M (p<0.001) and VIM (p<0.0001) were significantly upregulated in NPDR+DKD compared with NPDR. In ROC analysis, B2M and VIM could distinguish DKD from NPDR with area under the curve values of 0.9000 (p < 0.0001) and 0.9950. Our proteomic study revealed alterations in the proteomic profile and identified VIM and B2M as early biomarkers of DKD, laying the foundation for the prevention, diagnosis and treatment of DKD.

Progress and Challenges of Anti-VEGF Agents and Their Sustained-Release Strategies for Retinal Angiogenesis.

Currently, the treatment for ocular neovascular diseases, including diabetic macular edema (DME) and age-related macular degeneration (AMD), mainly involves repeated intravitreal injection of anti-vascular endothelial growth factor (VEGF) drugs. Although it can preserve vision, repeated injections are an invasive treatment modality, leading to serious complications and reducing patient adherence to treatment. To reduce the frequency of administration, prolong the time of drug action, and avoid repeated intravitreal injections, the combination of sustained-release materials with anti-VEGF drug therapy has become an emphasis in ophthalmology. In this review, we highlight the current state of anti-VEGF technology, its challenges, and the sustained-release strategies under investigation or being used in clinical practice. Both continuous release and considerable therapeutic effects can be achieved by encapsulating anti-VEGF drugs in sustained-release materials to minimize the number of intravitreal injections. At present, two sustained-release materials are being tested in clinical research, and although basic research shows the strong therapeutic application prospects of extended-release drugs, its challenges mainly involve the discrepancy between the release rates in vitro and the efficiency of the drugs in vivo. Briefly, sustained release of anti-VEGF agents is an advantageous strategy for treating retinal angiogenesis.

Influences of Rapamycin on Retinal Ganglion Cells in Rats with Acute High Intraocular Pressure Through Regulating COX-2.

The study aimed to explore the influences of rapamycin on the retinal ganglion cells in rats with acute high intraocular pressure through regulating cyclooxygenase-2 (COX-2). 36 Sprague-Dawley rats were randomly assigned to the normal group (n=12), model group (n=12) and rapamycin group (n=12). The rats in the normal group were normally fed, those in the model group were prepared the model of acute high intraocular pressure and injected with normal saline, and those in the rapamycin group were given rapamycin. At 7 d after the operation, sampling was performed. The expressions of COX-2 and Caspase-3 were detected via immunohistochemistry, and their protein expressions were determined using Western blotting (WB). Quantitative polymerase chain reaction (qPCR) was conducted to measure the messenger ribonucleic acid (mRNA) expression levels, and cell apoptosis was evaluated using terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) assay. The content of interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-α) was determined via enzyme-linked immunosorbent assay (ELISA). Compared with those in the normal group, the positive expression levels rose substantially in the other two groups, and those in the rapamycin group were notably lower (p<0.05). The relative protein expression levels in the model group and rapamycin group were higher, and the rapamycin group exhibited remarkable decreases (p<0.05). In comparison with the normal group, the other two groups had considerably raised relative mRNA expression levels and those in the rapamycin group were lower (p<0.05). The cells in the model and rapamycin groups had a higher apoptosis rate, and the apoptosis rate of cells in the rapamycin group was lower (p<0.05). Compared with that in the normal group, the content of IL-6 and TNF-α was elevated in the other two groups and their content in the rapamycin group was lower. Rapamycin inhibits COX-2 to repress inflammation and apoptosis, thereby exerting a protective effect on the retinal ganglion cells in rats with acute high intraocular pressure.

Circ-NCX1 inhibits LPS-induced chondrocyte apoptosis by regulating the miR-133a/SIRT1 axis.

Osteoarthritis (OA) is a degenerative joint disease, which is characterized by the degeneration of articular cartilage, thickening of subchondral bone, and inflammation of the synovial membrane. In this study, we aimed to investigate the effects and underlying mechanisms of circ-NCX1 in lipopolysaccharide (LPS)-induced injury in SW1353 chondrocytes, an in vitro model of OA. The levels of circ-NCX1, miR-133a, and related apoptotic proteins were determined by RT-qPCR. MTT assay was used to evaluate the cell viability. The apoptosis was determined by flow cytometry, whereas the expression of apoptosis proteins was detected by Western blot. Immunofluorescence was used to detect cleaved caspase-3 expression in cells. Luciferase reporter assay was used to verify the interaction between circ-NCX1 and miR-133a, and between miR-133a and Silent information regulator 2 homolog 1 (Sirt1). The results showed that the overexpression of circ-NCX1 significantly upregulated the chondrocyte viability and proliferation, and alleviated apoptosis in LPS-induced SW1353 cells. Immunofluorescence results showed that the overexpression of circ-NCX1 significantly reduced expression of LPS-stimulated cleaved Caspase-3. The RT-qPCR results showed that the overexpression of circ-NCX1 inhibited mRNA levels of cleaved Caspase-3 and Bax, and promoted mRNA levels of Bcl-2. Luciferase reporter assay showed that circ-NCX1 targeted miR-133a, and miR-133a directly targeted the Sirt1. In addition, overexpression of circ-NCX1 inhibited chondrocyte apoptosis and promoted Akt phosphorylation via the miR-133a/Sirt1 axis in LPS-induced chondrocytes. In conclusion, circ-NCX1 may serve as an important regulator of LPS-induced chondrocyte apoptosis through the miR-133a/Sirt1 axis, and may be involved in the development of OA.

Downregulation of fatty acid binding protein 4 alleviates lipid peroxidation and oxidative stress in diabetic retinopathy by regulating peroxisome proliferator-activated receptor γ-mediated ferroptosis.

This study aims to explore the role of fatty acid binding protein 4 (FABP4) in diabetic retinopathy (DR), and to elucidate the potential regulatory mechanism. We firstly developed a mouse model of DR by injection with streptozocin (STZ) into C57BL/6 male mice and a cell model of DR by induction of high glucose (HG) to ARPE-19 cells. BMS309403, an inhibitor of FABP4, was employed for treatment. The blood glucose in vivo was monitored and the histological changes of retinal tissues were observed by hematoxylin and eosin staining and Evans blue assay. The expression level of FABP4 was detected by western blot and Immunohistochemical staining. The critical factors related to lipid peroxidation and oxidative stress were detected using their commercial kits, respectively. Prussian blue staining, iron content assay and thiobarbituric acid-reactive substances (TBARS) assay were conducted to evaluate ferroptosis. As a result, FABP4 was elevated in retina and serum of STZ-induced mice and in HG-induced ARPE-19 cells. BMS309403 treatment notably alleviated reduced blood glucose, reduced histological damage, and vascular permeability. In addition, BMS309403 treatment inhibited lipid peroxidation, oxidative stress, and ferroptosis both in vivo and in vitro. Furthermore, BMS309403 promoted the activation of peroxisome proliferator-activated receptor γ (PPARγ). GW9662 (an inhibitor of PPARγ) or Erastin (an inducer of ferroptosis) partially weakened the suppressive effects of BMS309403 on HG-induced lipid peroxidation, oxidative stress and ferroptosis. Taken together, FABP4 inhibition alleviates lipid peroxidation and oxidative stress in DR by regulating PPARγ-mediated ferroptosis.

Human embryonic stem cell-derived extracellular vesicles alleviate retinal degeneration by upregulating Oct4 to promote retinal Müller cell retrodifferentiation via HSP90.

OBJECTIVE: Retinal degenerative diseases remain the dominant causes of blindness worldwide, and cell replacement is viewed as a promising therapeutic direction. However, the resources of seed cells are hard to obtain. To further explore this therapeutic approach, human embryonic stem extracellular vesicles (hESEVs) were extracted from human embryonic stem cells (hESCs) to inspect its effect and the possible mechanism on retinal Müller cells and retinal function. METHODS: hESEVs were extracted by multi-step differential centrifugation, whose morphologies and specific biomarkers (TSG101, CD9, CD63, and CD81) were observed and measured. After hESEVs were injected into the vitreous cavity of RCS rats, the retinal tissues and retinal functions of rats were assessed. The alteration of Müller cells and retinal progenitor cells was also recorded. Microvesicles (MVs) or exosomes (EXOs) were extracted from hESCs transfected with sh-HSP90 or pcDNA3.1-HSP9, and then incubated with Müller cells to measure the uptake of EVs, MVs, or EXOs in Müller cells by immunofluorescence. The retrodifferentiation of Müller cells was determined by measuring Vimentin and CHX10. qRT-PCR and western blot were used to detect HSP90 expression in MVs and evaluate Oct4 level in Müller cells, and Co-IP to inspect the interaction of HSP90 and Oct4. RESULTS: RCS rats at the postnatal 30 days had increased retinal progenitor cells which were dedifferentiated from Müller cells. hESEVs were successfully extracted from hESCs, evidenced by morphology observation and positive expressions of specific biomarkers (TSG101, CD9, CD63, and CD81). hESEVs promoted Müller cells dedifferentiated and retrodifferentiated into retinal progenitor cells evidenced by the existence of a large amount of CHX10-positive cells in the retinal inner layer of RCS rats in response to hESEV injection. The promotive role of hESEVs was exerted by MVs demonstrated by elevated fluorescence intensity of CHX10 and suppressed Vimentin fluorescence intensity in MVs rather than in EXOs. HSP90 in MVs inhibited the retrodifferentiation of Müller cells and suppressed the expression level of Oct4 in Müller cells. Co-IP revealed that HSP90 can target Oct4 in Müller cells. CONCLUSION: hESEVs could promote the retrodifferentiation of Müller cells into retinal progenitor cells by regulating the expression of Oct4 in Müller cells by HSP90 mediation in MVs.

Silence of TPPP3 suppresses cell proliferation, invasion and migration via inactivating NF-κB/COX2 signal pathway in breast cancer cell.

Malignant phenotypes are leading causes of death in patients with breast cancer (BC). Previously, it has been proved that tubulin polymerization promoting protein 3 (TPPP3) participates in cell progressions in several human cancers. Little is known about the functions of TPPP3 in BC. Herein, we detected the expression of TPPP3 in 54 clinical BC tissues and two BC cell lines by immunohistochemistry and Western blot. CCK-8, wound healing, colony formation and Transwell assays were used to assess cell proliferation, clone formation, invasion and migration of MCF-7 and T47D cells after transfection with TPPP3 siRNA. Meanwhile, related-proteins expression was detected using Western blot. TPPP3 was found to be highly expressed in the tissues from the patients with BC. Poor outcomes were associated with the high expression of TPPP3 in all patients with BC. When MCF-7 and T47D cells receiving TPPP3 siRNA transfection, the capacities of proliferation, clone formation, invasion and migration were suppressed and the expression of MMP-2/-9 and NF-κB p65/COX2 was notably reduced. The dual-luciferase reporter assay indicated that the promoter regions of NF-κB p65 could combine to TPPP3. Overall, the present study demonstrated that TPPP3 played a significant role in BC, and its inhibition lead to the suppression of NF-κB/COX-2 signalling pathway along with the reduction of malignant phenotypes. SIGNIFICANCE OF THIS STUDY: Previously, it has been proved that tubulin polymerization promoting protein 3 (TPPP3) participates in cell progression in several human cancers. Little is known about the function of TPPP3 in BC. Our study was the first direct evidence to support the role of TPPP3 in tumorigenesis and metastasis of BC. Although the underlying mechanism has not been fully delineated, these findings suggested that TPPP3 was an important factor in the tumour progression and metastasis of BC cells and provided a molecular basis for potential therapeutic implications in the treatment of patients with BC.

Exosomes derived from RPE cells under oxidative stress mediate inflammation and apoptosis of normal RPE cells through Apaf1/caspase-9 axis.

This study aims to explore the effects of exosomes, secreted by retinal pigment epithelial (RPE) cells under oxidative stress (OS), on apoptosis and inflammation of normal RPE cells. Exosomes secreted by normal RPE cells (named as exo) and rotenone (2.5 µmol/L) stimulated RPE cells (named as rot-exo) were isolated and extracted by multi-step differential centrifugation for morphology observation under a transmission electron microscopy. pcDNA3.1a, pcDNA3.1a-Apaf1, and p3xFlag-CMV-caspase-9 plasmids were constructed and transfected into ARPE-19 cells. Exosomes secreted by ARPE-19 cells were injected into the vitreous body of rats to verify the effect of Apaf1 and caspase-9 on cell apoptosis and inflammation. Co-immunoprecipitation was applied to clarify the interaction of Apaf1 with caspase-9. Exosomes secreted by rotenone stimulated ARPE-19 cells could induce cell apoptosis, oxidative injury, and inflammation in ARPE-19 cells. Exosomes secreted under OS can damage retinal functions of rats and have upregulated expression of Apaf1. Overexpression of Apaf1 in exosomes secreted under OS can cause the inhibition of cell proliferation, the increase of cell apoptosis and elicitation of inflammatory response in ARPE-19 cells. Exosomes derived from ARPE-19 cells under OS regulate Apaf1 expression to increase cell apoptosis and to induce oxidative injury and inflammatory response through a caspase-9 apoptotic pathway.

Profiling of glucosinolates and flavonoids in Rorippa indica (Linn.) Hiern. (Cruciferae) by UHPLC-PDA-ESI/HRMS(n).

An UHPLC-PDA-ESI/HRMS(n) profiling method was used to identify the glucosinolates and flavonoids of Rorippa indica (Cruciferae), a wild vegetable and Chinese herb used to treat cough, diarrhea, and rheumatoid arthritis. Thirty-three glucosinolates, more than 40 flavonol glycosides, and 18 other phenolic and common organic compounds were identified. The glucosinolates and polyphenols were separated by UHPLC. High-resolution deprotonated molecules provided high accuracy mass values that were used to determine formulas and provide putative identification of the glucosinolates and flavonoids. The fragments from multistage mass spectrometry were used to elucidate the structures. The concentrations of the main components were based on UV peak areas and molar relative response factors with a single calibration standard. This study found this plant to be a rich source for glucosinolates, containing 24 new glucosinolates, including 14 glucosylated glucosinolates that were previously unidentified.