Experimental Study on the Application of Bioactive Xenogeneic Porcine Cancellous Bone for Cervical Intervertebral Fusion in Goats.

STUDY DESIGN: An experimental study in a cervical intervertebral fusion goat model. OBJECTIVE: To investigate the effect of bioactive xenogeneic porcine cancellous bone applied to the intervertebral fusion of goat cervical vertebrae. SUMMARY OF BACKGROUND DATA: Although autogenous bone achieves satisfied outcome in cervical intervertebral fusion, it is limited and cause several complications. The application of xenogeneic bone has potential to solve these problems. METHODS: Thirty local goats were randomly divided into 3 groups: group A (12 goats): autogenous tricortical iliac bone group; group B (6 goats): polyetheretherketone (PEEK) cage with autologous bone; and group C (12 goats): PEEK cage with bioactive xenogeneic porcine cancellous bone. C3-C4 discectomy was performed in each group and the above bone graft and bone graft substitutes were implanted. Lateral cervical spine x-rays were taken at preoperative; immediately postoperative; and 4, 8, 12, and 24 weeks postoperatively every goat. Disc space heights (DSHs) were measured on lateral x-rays. CT examination was performed at 12 and 24 weeks after surgery for the fusion score. After 4 and 8 weeks after surgery, 3 goats were euthanized in both groups A and C to evaluate the immune rejection response through histology. At 12 and 24 weeks after surgery, 3 goats were euthanized in each group. The cervical implants fusion outcome was evaluated through specimen histology observation. RESULT: As time extended, the immune rejection of bioactive xenogeneic porcine cancellous bone gradually subsided. Radiology, specimen observation, and histology manifested that the C3-4 vertebral bodies of goats in each group gradually fused. All the goats in each group achieved bony fusion at 24 weeks after surgery. In terms of preventing intervertebral space collapse, the PEEK cage could achieve better results. There was no significant difference in the remaining experimental data (P>0.05). CONCLUSIONS: Bioactive xenogeneic porcine cancellous bone can obtain satisfied fusion outcomes in cervical intervertebral fusion and is an ideal intervertebral fusion material in goats.

Resurfacing of multiple adjacent defects with free multipaddle SCIAP flaps.

BACKGROUND: The efficient resurfacing of multiple adjacent defects (MADs) requires precise reconstructive strategy. Various approaches (e.g., several flap transferring or prelamination of the recipient site) have been reported, but recipient-site impairments, pain, long hospitalization, and low cost-benefit results fatefully considered them as compromise approaches. This study aims to evaluate the feasibility of MADs reconstruction with free multipaddle superficial circumflex iliac artery perforator (SCIAP) flaps. METHODS: From Dec 2015 to Dec 2020, we enrolled patients with upper and lower extremity defects treated with various multipaddle SCIAP flaps (2-paddle, 3-paddle, and 4-paddle). Patient demographics and outcomes of each group were collected. RESULTS: Thirty-two, 21, and 6 patients underwent 2-paddle, 3-paddle, and 4-paddle SCIAP flaps transfers, respectively. All multipaddle SCIAP flaps survived without vascular problems, and the donor sites were closed directly. Except for 3 cases of 2-paddle SCIAP flaps drained by superficial circumflex iliac vein venous return, most cases (n = 56) were drained by venae comitans. Minor complications, including partial flap necrosis (4 cases) and lateral femoral cutaneous nerve palsies (11 cases), were treated conservatively. All patients were satisfied with the reconstructive outcome. CONCLUSION: Multiple adjacent defects reconstruction is still a Gordian knot and lacks a golden standard. The free multipaddle SCIAP flap was demonstrated as a promising alternative, not only enriching its versatility but also initially highlighting the "replace need with need" reconstructive demand.

Suicide among lymphoma patients.

BACKGROUND: Higher suicide rates were observed in patients diagnosed with lymphoma. In this study, we accurately identified patients with high-risk lymphoma for suicide by constructing a nomogram with a view to effective interventions and reducing the risk of suicide. METHODS: 235,806 patients diagnosed with lymphoma between 2000 and 2020 were picked from the Surveillance, Epidemiology, and End Results (SEER) database and randomly divided into training (N = 165,064) and validation set (N = 70,742). A combination of the Least absolute shrinkage and selection operator (LASSO) and Cox proportional hazards regression identified the predictors that constructed the nomogram. To assess the discrimination, calibration, clinical applicability, and generalization of this nomogram, we implemented receiver operating characteristic curves (ROC), calibration curves, decision curve analysis (DCA), and internal validation. The robustness of the results was assessed by the competing risks regression model. RESULTS: Age at diagnosis, gender, ethnicity, marital status, stage, surgery, radiotherapy, and annual household income were key predictors of suicide in lymphoma patients. A nomogram was created to visualize the risk of suicide after a lymphoma diagnosis. The c-index for the training set was 0.773, and the validation set was 0.777. The calibration curve for the nomogram fitted well with the diagonal and the clinical decision curve indicated its clinical benefit. LIMITATION: The effects of unmeasured and unnoticed biases and confounders were difficult to eliminate due to retrospective studies. CONCLUSION: A convenient and reliable model has been constructed that will help to individualize and accurately quantify the risk of suicide in patients diagnosed with lymphoma.

Immune checkpoint inhibitors as the second-line treatment for advanced esophageal squamous cell carcinoma: a cost-effectiveness analysis based on network meta-analysis.

BACKGROUND: Immune checkpoint inhibitors (ICIs) have demonstrated superior clinical efficacy in prolonging overall survival (OS) as the second-line treatment for advanced or metastatic esophageal squamous cell carcinoma (ESCC), and were recommended by the guidelines. However, it remains uncertain which ICI is the most cost-effective. This study assessed the cost-effectiveness of ICIs as the second-line treatment for ESCC based on the perspective of the Chinese healthcare system. METHODS: A network meta-analysis (NMA) was performed to obtain the Hazard ratios (HRs) for indirect comparisons. A three-state Markov model with a 10-year time horizon was conducted to assess the cost-effectiveness. The state transition probabilities were calculated with Kaplan-Meier (KM) curves data from clinical trial and HRs from the NMA. Utilities and costs were derived from local charges or previously published studies. Univariate and probabilistic sensitivity analyses (PSA) were performed to examine model robustness. The results were assessed based on the total costs, quality-adjusted life years (QALYs), and incremental cost-effectiveness ratios (ICERs). RESULTS: Five clinical trials (ATTRACTION-3, ESCORT, KEYNOTE-181, ORIENT-2, RATIONALE-302) with a total of 1797 patients were included in the NMA. The NMA showed that both camrelizumab and tislelizumab received relatively high rankings for progression-free survival (PFS) and OS. Compared with sintilimab, treatment with tislelizumab and camrelizumab gained 0.018 and 0.034 additional QALYs, resulting in incremental ICERs of $75,472.65/QALY and $175,681.9/QALY, respectively. Nivolumab and pembrolizumab produced lower QALYs and greater costs, suggesting that both were dominated in comparison to sintilimab. HRs and health state utilities were the most influential parameters in most univariate sensitivity analyses of paired comparisons. PSA results suggested that sintilimab had an 84.4% chance of being the most cost-effective treatment regimen at the WTP threshold of $38,223.34/QALY. In the scenario analysis, sintilimab would no longer be cost-effective, if the price of camrelizumab was assumed to decrease by 64.6% or the price of tislelizumab was assumed to decrease by 16.9%. CONCLUSIONS AND RELEVANCE: Among the five potential competing ICIs, sintilimab was likely to be the most cost-effective regimen as the second-line treatment for locally advanced or metastatic ESCC in China.

Efficacy of Sacubitril-Valsartan on Survival and Cardiac Remodeling in Hypotensive Heart Failure With Reduced Ejection Fraction: A Multicenter Study.

OBJECTIVE: To investigate whether hypotensive patients diagnosed with heart failure and reduced ejection fraction (HFrEF) might benefit from angiotensin receptor-neprilysin inhibitors (ARNis) in real-world practice because patients with baseline systolic blood pressure (SBP) of less than 100 mm Hg have been excluded from landmark trials. PATIENTS AND METHODS: In this multicenter study conducted between January 1, 2013, and December 31, 2021, a total of 7562 symptomatic patients with HFrEF were enrolled and grouped by SBP (hypotension was defined as an SBP of less than 100 mm Hg) and ARNi use as follows: group 1, hypotensive/non-ARNi users (n=484); group 2, hypotensive/ARNi users (n=308); group 3, nonhypotensive/non-ARNi users (n=4560); and group 4, nonhypotensive/ARNi users (n=2210). Inverse probability of treatment weighting was used to balance baseline characteristics for survival analysis. RESULTS: Diverse baseline characteristics and lower rates of medication use were found among non-ARNi users compared with ARNi users. Hypotensive/ARNi users had lower ARNi initiation doses than nonhypotensive/ARNi users. We observed significantly lower mortality, composite heart failure hospitalization, and CV death for hypotensive/ARNi and the other 2 nonhypotensive groups (groups 3 and 4) during a median follow-up of 3.43 years (all P<.05), with a similar effect on reverse remodeling for the hypotensive/ARNi group compared with the hypotensive/non-ARNi group. The event-free survival benefits of ARNi vs renin-angiotensin system inhibitors were consistent with the lower boundary of SBP for clinical benefits found until 88 mm Hg (spline curves) after inverse probability of treatment weighting. CONCLUSION: Patients with HFrEF and hypotension may still benefit from ARNi treatment. Patients with hypotensive HFrEF should not be routinely excluded from ARNi use in a real-world setting.

Experimental Study of Skin Contraction Induced by Bipolar Radiofrequency.

Background: Facial skin relaxation has become an important part in solving the problem of facial rejuvenation. Minimally invasive or noninvasive skin-tightening procedures have become a trend for facial rejuvenation. Bipolar radiofrequency (RF) is a new option for treating skin relaxation and is more effective than noninvasive surgery without surgical incision. Objective: To explore the effect of different bipolar RF powers on the area of the original box, changes of skin and subcutaneous tissue thickness and numbers of fibroblasts in rabbits. Design: The research team performed an animal study. Setting: This study took place in Affiliated Hospital of Nanjing University of Chinese Medicine. Participants: Eighteen common-grade adult New Zealand rabbits (female, 2.5-3.0 kg). Methods: Bipolar radiofrequency therapy was given to a girl rabbit on the left side of the treatment area. Standard HE and Masson staining were performed to assess the pathological changes, area of the original box and the number of fibroblasts in skin and subcutaneous tissues. Outcome Measures: (1) The area of the original box, changes of skin and subcutaneous tissue thickness, and numbers of fibroblasts under different bipolar RF temperatures or under different bipolar RF powers immediately after surgery, 1 month after surgery and 3 months after surgery were observed. (2) Standard HE and Masson staining results. Results: Under the condition of certain instrument power, at 36de 38d and 40nd the area of the original box shrank to different degrees immediately after surgery (16.54±0.37, 17.78±0.03, 17.19±0.01), 1 month after surgery (16.59±0.31, 17.82±0.01, 18.34±0.30) and 3 months after surgery (16.89±0.12, 18.16±0.14, 19.23±0.32) compared with that before surgery (P < .05). Under specific temperature conditions, at 16 W, 18 W, 20 W, and 22 W, the area of the original box shrank to different degrees immediately after surgery (16.40±0.49, 15.55±0.57, 17.54±0.12, 16.19±0.27), 1 month after surgery (16.88±0.12, 17.46±0.02, 18.05±0.35, 19.41±0.08) and 3 months after surgery (19.09±1.01, 18.30±0.69, 20.00±0.29, 21.20±0.90) compared with that before surgery (P < .05). When the power was fixed, the thickness of skin and subcutaneous tissue decreased immediately after surgery (6.7, 6.8, 7), 1 month after surgery (6, 6.1, 6.3) and 3 months after surgery (6.4, 6.5, 6.2) at different temperatures (P < .05). When the temperature was fixed, the thickness of skin and subcutaneous tissue decreased immediately after surgery (6.1, 6.08, 6.03), 1 month after surgery (6.2, 6.15, 6.13), and 3 months after surgery (6.2, 6.23, 6.03) under different powers (P < .05). Under the condition of certain instrument power, at 36de 38d and 40n, the number of fibroblasts increased to different degrees immediately after surgery (26.54±2.37, 30.78±3.03, 37.19±4.01), 1 month after surgery (28.59±2.31, 34.82±3.01, 40.34±4.30), and 3 months after surgery (30.89±0.12, 38.16±0.14, 42.23±0.32) compared with that before surgery, and all were statistically significant (P < .05). Under specific temperature conditions, at 16 W, 18 W, 20 W, and 22 W, the number of fibroblasts increased to different degrees immediately after surgery (28.29±2.49, 30.97±3.57, 38.74±3.12, 45.68±4.27), 1 month after surgery (30.88±3.12, 32.46±4.02, 41.05±0.35, 50.41±0.08), and 3 months after surgery (29.99±2.01, 33.30±2.69, 39.00±3.29, 23.20±2.90) compared with that before surgery, and all were statistically significant (P < .05). Conclusions: Our study clarifies that bipolar RF can decrease the skin and subcutaneous tissue thickness and increase the numbers of fibroblasts at the temperature of 36°C, 38°C, and 40°C and frequency of 16-22 W, which has a therapeutical effect on skin contraction. Our study might effectively improve the skin slack of patients, and the postoperative maintenance rate is high, and will not cause obvious complications. This study may provide a theoretical direction for clinicians to tighten the skin of patients using bipolar RF.

Single-cell and spatial architecture of primary liver cancer.

Primary liver cancer (PLC) poses a leading threat to human health, and its treatment options are limited. Meanwhile, the investigation of homogeneity and heterogeneity among PLCs remains challenging. Here, using single-cell RNA sequencing, spatial transcriptomic and bulk multi-omics, we elaborated a molecular architecture of 3 PLC types, namely hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (ICC) and combined hepatocellular-cholangiocarcinoma (CHC). Taking a high-resolution perspective, our observations revealed that CHC cells exhibit internally discordant phenotypes, whereas ICC and HCC exhibit distinct tumor-specific features. Specifically, ICC was found to be the primary source of cancer-associated fibroblasts, while HCC exhibited disrupted metabolism and greater individual heterogeneity of T cells. We further revealed a diversity of intermediate-state cells residing in the tumor-peritumor junctional zone, including a congregation of CPE+ intermediate-state endothelial cells (ECs), which harbored the molecular characteristics of tumor-associated ECs and normal ECs. This architecture offers insights into molecular characteristics of PLC microenvironment, and hints that the tumor-peritumor junctional zone could serve as a targeted region for precise therapeutical strategies.

Role of endoscopic third ventriculostomy in patients undergoing resection of pulvinar area lesions: Preliminary clinical results.

BACKGROUND: Patients with pulvinar area lesions may develop hydrocephalus at any stage. The role of endoscopic third ventriculostomy (ETV) in this setting remains unclear. METHOD: We retrospectively enrolled 15 patients with a mean age of 43 years who underwent endoscopic resection of pulvinar area lesions using the supracerebellar infratentorial approach (SCITA). We compared the different modalities of hydrocephalus management and their outcomes. RESULTS: Nine of 15 patients (60.0%) had preoperative obstructive hydrocephalus. Five patients underwent ETV before tumor resection, and none developed postoperative hydrocephalus. Four patients underwent one-stage surgery for tumor removal, and one patient with a polymorphous low-grade neuroepithelial tumor of the young required postoperative ETV. Another patient with diffuse astrocytoma and hydrocephalus underwent concurrent lamina terminalis fenestration and endoscopic resection via the SCITA, which resulted in the resolution of hydrocephalus. The preoperative ETV group had no major postoperative complications, while the non-ETV group had three (0/5 vs. 3/4, P = 0.048). The ETV group also had a shorter intensive care unit stay; however, the difference was not significant (1.2 vs. 2.8; P = 0.188). ETV was effective in alleviating symptoms of postoperative hydrocephalus in patients with midbrain-invading tumors. CONCLUSION: Endoscopic surgery via the SCITA can address both tumor and hydrocephalus issues in some cases but has a higher surgical risk and postoperative hydrocephalus rate. Preoperative ETV can prevent these complications and improve postoperative outcomes.

Identification of sex pheromone of red swamp crayfish Procambarus clarkii and exploration of the chemosensory mechanism of their antennae.

Red swamp crayfish, Procambarus clarkii, is a globally invasive species, which has caused great damage to biodiversity, agriculture, and fishing. Therefore, the development of effective management methods, such as pheromone control, is necessary for biological control and biodiversity protection. However, the components of P. clarkii sex pheromones have not yet been explored, and the chemosensory mechanism of the P. clarkii antennae after stimulation by sex pheromone also remains unknown. In this study, we isolated and identified the candidate bioactive component of the female P. clarkii sex pheromone using ultrafiltration centrifugation, semi-preparative liquid phase separation and omics technologies and conducted bioassays to determine its attraction ability. Meanwhile, RNA-Seq technology was used to analyze the potential chemosensory mechanism of antennae. Our results indicated that the male P. clarkii were uniaxially attracted to the female crude conditioned water (FCW), medium fraction (MF, isolated by ultrafiltration centrifugation), and preparative fragment 6 of females (PFF6, isolated by semi-preparative liquid phase separation). Metabolomic analysis revealed the presence of 18 differential metabolites between the PFF6 and PFM6 samples, among which 15 were significantly upregulated in the PFF6 sample. Bioassay test also showed that mestranol, especially at concentrations of 10-5-10-2 mol∙l-1, could significantly attract P. clarkii males; therefore, mestranol was identified as the candidate sex pheromone component of P. clarkii females. Furthermore, RNA-Seq results showed that most differentially expressed genes (DEGs) enriched in lipid metabolism and signal transduction pathways were up-regulated in P. clarkii males. In addition, high expressions of Ca2+-binding protein and ion transporting ATPases may enhance the sensitivity of the antennae of P. clarkii males towards sex pheromones. Our study provides data on P. clarkii sex pheromone composition and reveals the molecular mechanism of sex pheromone response in P. clarkii. Moreover, our study provides a referable method for the isolation of candidate bioactive molecules from the P. clarkii sex pheromone.

LncRNA Gm44206 Promotes Microglial Pyroptosis Through NLRP3/Caspase-1/GSDMD Axis and Aggravate Cerebral Ischemia-Reperfusion Injury.

Inhibition of the inflammatory response triggered by microglial pyroptosis inflammatory activation may be one of the effective ways to alleviate cerebral ischemia-reperfusion injury, the specific mechanism of which remains unclear. In this study, BV-2 microglia with or without oxygen-glucose deprivation/reoxygenation (OGD/R) or long noncoding RNA (lncRNA) Gm44206 knockdown were used as cell models to conduct an in vitro study. Detection of lactate dehydrogenase release and pyroptosis-related protein levels was performed using a corresponding kit and western blotting, respectively. Proliferation of microglia was evaluated by CCK8 assay. Enzyme-linked immunosorbent assay was applied for measuring levels of proinflammatory cytokines. This study verified the involvement of microglial pyroptosis as well as upregulation of NLRP3, Caspase-1, GSDMD, and Apoptosis-associated Speck-like protein containing a C-terminal caspase-recruitment domain (ASC) in cerebral ischemia-reperfusion injury. Moreover, knockdown of lncRNA Gm44206 could alleviate OGD/R-induced microglial pyroptosis and cell proliferation inhibition through the NLRP3/Caspase-1/GSDMD pathway, thus decreasing the release of proinflammatory cytokines, including interleukin (IL)-1ß, IL-6, IL-18, and tumor necrosis factor-alpha. In conclusion, this study established a correlation between microglial pyroptosis and cerebral ischemia-reperfusion injury and identified lncRNA Gm44206 as a potential regulator of NLRP3/Caspase-1/GSDMD axis-mediated microglial pyroptosis, which could be considered a promising therapeutic target.

Study of platelet-rich fibrin promoting endothelial cell differentiation and angiogenesis induced by transplantation of adipose-derived stem cells.

Diabetic patients are characterized by long wound healing time, and adipose stem cells (ADSCs) can secrete growth factors to promote angiogenesis and improve diabetic wound healing. In this research, we attempted to interrogate the impact of platelet-rich fibrin (PRF) on ADSCs in diabetic wound healing. ADSCs were harvested from human adipose tissues and identified through flow cytometry. After pretreatment with cultured medium supplemented with different concentrations of PRF (2.5%, 5%, and 7.5%), proliferation and differentiation capacity of ADSCs were assessed by CCK-8 assay, qRT-PCR and immunofluorescence (IF), respectively. Tube formation assay measured angiogenesis. Western blot analysis analyzed expression of endothelial markers and the extracellular signal-regulated kinase (ERK) and serine/threonine kinase (Akt) pathways in PRF-induced ADSCs. The CCK-8 experiment indicated that PRF enhanced proliferation of ADSCs in dose-dependent manner, relative to normal control group. The expression of endothelial markers and the capacity of tube formation were significantly promoted by 7.5% PRF. The release of growth factors containing vascular endothelial grow factor (VEGF) and insulin-like growth factor-1 (IGF-1) from PRF was increased with the extension of detection time. When the receptors of VEGF or/and IGF-1 were neutralized, ADSCs differentiation into endothelial cells were obviously inhibited. Additionally, PRF stimulated ERK and Akt pathways, and the inhibitors of ERK and Akt attenuated PRF-induced differentiation of ADSCs into endothelial cells. In conclusion, PRF promoted endothelial cell differentiation and angiogenesis induced by ADSCs in diabetic wound healing, which appears to give guidance for treating patients.

Case report: Medulla oblongata and cervical cord reperfusion injury after intracranial vertebral artery angioplasty and stenting.

Background: White cord syndrome is an uncommon complication characterized by delayed neurologic deterioration with no other identified cause after spinal decompression surgery. Its etiology is attributed to spinal cord reperfusion injury. Here, we present the first case of an extended version of white cord syndrome, with concomitant involvement of the medulla oblongata and cervical cord reperfusion injury after intracranial vertebral artery angioplasty and stenting. Case presentation: A 56-year-old male suffered an ischemic stroke in the right anteromedial medulla oblongata. Angiography revealed bilateral vertebral artery stenosis in the intracranial segment. We performed elective left vertebral artery angioplasty and stenting. An intraoperative flow arrest in the left VA occurred and was stopped after the withdrawal of the catheter. Several hours after the operation, the patient developed occipital headache, back neck pain, dysarthria, and worsening left-sided hemiplegia. Magnetic resonance imaging revealed hyperintensity and swelling in the medulla oblongata and cervical cord, in addition to small medullary infarction. A digital subtraction angiography revealed intact vertebrobasilar arteries and patency of the left vertebral artery, left posterior inferior cerebellar artery, and implanted stent. We considered that the reperfusion injury had caused the complication. After treatment, the patient's symptoms and neurologic deficits greatly improved. He achieved a favorable outcome at the 1-year follow-up, with normal intensity restored in the medulla oblongata and cervical cord on magnetic resonance imaging. Conclusion: Concomitant reperfusion injury in the medulla oblongata and cervical cord secondary to vertebral artery angioplasty and stenting is extremely rare. However, this potentially devastating complication requires early recognition and prompt treatment. Maintaining the antegrade flow during vertebral artery endovascular treatment is a precaution against reperfusion injury.

FOXK1 regulates epithelial-mesenchymal transition and radiation sensitivity in nasopharyngeal carcinoma via the JAK/STAT3 signaling pathway.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is the most common head and neck tumor in China. Forkhead box (FOX) proteins have 19 subfamilies, which can maintain cell metabolism, regulate cell cycle and cell growth, etc. FOXK1 is a member of the FOX family, and studies have found that FOXK1 is closely related to tumors. OBJECTIVE: This experiment aims to study the effects of FOXK1 interference on proliferation, apoptosis, invasion, epithelial-mesenchymal transition (EMT), and radiosensitivity, by regulating the Janus kinas/signal translator and activator of the transfer 3 (JAK/STAT3) pathway. METHODS: The expression of FOXK1 was detected via immunohistochemistry using clinical nasopharyngeal carcinoma tissues and adjacent tissues. The relationship between FOXK1 expression and tumor stage was subsequently evaluated. The colony formation rate was calculated through the colony formation experiment. Cell apoptosis and cell cycle distribution were detected using flow cytometry, while cell invasion was detected using the Transwell method. The number of cells in the nucleus of each group after 30 min, 4 h, and 24 h of radiotherapy with the 2 Gy dose was counted using immunofluorescence under γ-H2AX focal points of a laser confocal microscope. RESULTS: FOXK1 is clearly expressed in the patients' cancer tissues. The expression of FOXK1 was significantly correlated with the patient's sex. FOXK1 interference or Peficitinib can upregulate the apoptosis rate of 5-8 F and CNE-2 cells; increase the G2 phase of cells; and inhibit the invasion, migration, and EMT of cells. At the same time, FOXK1 interference can downregulate the protein expression of p-JAK1, p-JAK2, and p-STAT3 in cells. Interference from FOXK1 or Peficitinib alone can reduce the rate of cell colony formation under different radiation doses, and enhance the green fluorescence intensity of γ-H2AX in the nucleus after 4 and 24 h of the 2 Gy dose of radiotherapy. These results are optimal when FOXK1 interference and Peficitinib are used together. CONCLUSION: FOXK1 interference in NPC cells can regulate EMT through the JAK/STAT3 signal pathway, enhance the radiosensitivity of cells, and thus inhibit tumor cell progression.

Levosimendan attenuates electrical alternans and prevents ventricular arrhythmia during therapeutic hypothermia in isolated rabbit hearts.

BACKGROUND: Therapeutic hypothermia (TH) increases the susceptibility to ventricular arrhythmias (VAs) by prolonging action potential duration (APD) and facilitating arrhythmogenic spatially discordant alternans (SDA). Levosimendan, a calcium sensitizer, has been reported to shorten APD by enhancing the adenosine triphosphate (ATP)-sensitive K current. OBJECTIVE: The purpose of this study was to test the hypothesis that, during TH, levosimendan shortens the already prolonged APD, attenuates SDA, and prevents VA. METHODS: Langendorff-perfused isolated rabbit hearts were subjected to TH (30°C) for 15 minutes, followed by treatment with either levosimendan 0.5 µM (n = 9) or vehicle (n = 8) for an additional 30 minutes under TH. Using an optical mapping system, epicardial APD was evaluated by S1 pacing. SDA threshold was defined as the longest pacing cycle length (PCL) that induces the phenomenon of SDA. Ventricular fibrillation (VF) inducibility was evaluated by burst pacing for 30 seconds at the shortest PCL that achieved 1:1 ventricular capture. RESULTS: During TH, levosimendan shortened ventricular APD (PCL 400 ms; from 259 ± 8 ms to 241 ± 18 ms; P = .036) and decreased SDA threshold (from 327 ± 88 ms to 311 ± 68 ms; P = .011). VF inducibility was lowered from 39% ± 30% to 14% ± 12% with levosimendan (P = .018), whereas APD at PCL 400 ms (P = .161), SDA threshold (P = 1), and VF inducibility (P = .173) were not changed by vehicle. CONCLUSION: During TH, levosimendan could protect hearts against VA by shortening APD and decreasing SDA threshold. Enhancing ATP-sensitive K current with levosimendan might be a novel approach to preventing VA during TH.

Real-World Experience of Angiotensin Receptor-Neprilysin Inhibition in Reduced Ejection Fraction Heart Failure Patients With Advanced Kidney Disease.

OBJECTIVE: To investigate the effectiveness and safety of angiotensin receptor-neprilysin inhibitors (ARNIs) in real-world patients with heart failure with reduced ejection fraction (HFrEF) and advanced chronic kidney disease (estimated glomerular filtration rate [eGFR] < 30 mL/min per 1.73 m2), which have been excluded from the landmark trials. PATIENTS AND METHODS: This study examined 3281 patients pooled from two multicenter HFrEF cohorts, and 661 patients with baseline eGFR less than 30 mL/min per 1.73 m2 were further analyzed (the Taiwan Society of Cardiology - Heart Failure with reduced Ejection Fraction (TSOC-HFrEF) registry: May 1, 2013 to October 31, 2014, and the Treatment with Angiotensin Receptor neprilysin inhibitor fOr Taiwan Heart Failure patients (TAROT-HF) study: March 1, 2017, to December 31, 2018). Propensity score matching was performed to adjust for confounders. At 1-year follow-up, all-cause mortality, total heart failure hospitalizations, renal function, and left ventricular ejection fraction (LVEF) were used as the endpoints. RESULTS: After propensity score matching, 510 patients (age, 69.8±13.9 years; male, 61.0%; mean LVEF, 29.8±7.3%; mean eGFR, 19.8±9.0 mL/min per 1.73 m2) were included in the final analysis, including 278 patients receiving ARNI treatment (ARNI group) and 232 patients not on ARNI treatment (non-ARNI group). Baseline characteristics were comparable between the two groups. At 1 year, eGFR and LVEF measurements were significantly higher in the ARNI group than in the non-ARNI group (25.0±17.1 mL/min per 1.73 m2 vs 21.4±17.5 mL/min per 1.73 m2; P=.04; and 40.1±12.9% vs. 33.1±10.8%, P<.001, respectively). The ARNI group had significantly lower risks of 1-year all-cause mortality (19.4 vs 30.9 per 100-person year; P=.02), and total HF rehospitalizations (70.0 vs 110.4 per 100-person year; P=.01) than non-ARNI users. CONCLUSION: Our results show the effectiveness of ARNIs in HFrEF patients with advanced chronic kidney disease in a real-world setting.

A ten long noncoding RNA-based prognostic risk model construction and mechanism study in the basal-like immune-suppressed subtype of triple-negative breast cancer.

Background: According to the Fudan University Shanghai Cancer Center (FUSCC) system, triple-negative breast cancer (TNBC) is divided into four stable subtypes: (I) luminal androgen receptor, (II) immunomodulatory, (III) basal-like immune-suppressed (BLIS), and (IV) mesenchymal-like. However, the treatment outcomes of the corresponding targeted therapies are unsatisfactory, especially for the BLIS subtype. Therefore, we aimed to identify the key long noncoding RNAs (lncRNAs) to construct a prognostic model for BLIS subtype and discover potential targets to explore potential therapeutic strategies in this study. Methods: The FUSCC cohort was used to establish a prognostic risk model via least absolute shrinkage and selection operator (LASSO) and Cox regression analysis. The Cancer Genome Atlas (TCGA) cohort was then used to evaluate and verify the model. To understand the functional aspects of the model, functional, immune landscape, mutation, and drug sensitivity analyses were performed between high- and low-risk groups. Results: Ten prognostic-related lncRNAs identified, including C5ORF66-AS2, DIO3OS, FZD10-DT, LINC00393, LNC-ERI1-32, LNC-FOXO1-2, LNC-SPARCL1-1, HCG23, LNC-MMD-4 and LNC-TMEM106C-6, were selected for risk score system construction. The results showed that the model constructed could divide the patients with BLIS subtype into two groups of high and low risk, and patients with higher risk scores had shorter recurrence-free survival. In addition, drug sensitivity analysis identified 3 compounds, including BMS-754807, cytochalasin b, and linifanib, that could have a potential therapeutic effect on patients with the BLIS subtype. Conclusions: The risk prognosis model showed good prognostic value for the BLIS subtype patients, and the ten lncRNAs may be potential therapeutic targets.

[The destructive role of soluble Robo4 secreted by the M1-polarized-microglia during cerebral ischemia-reperfusion in blood-brain barrier integrity].

This project was aimed to investigate the role and the underlying mechanism of microglia polarization on blood-brain barrier (BBB) during cerebral ischemia-reperfusion. After construction of the mouse model of cerebral ischemia-reperfusion, upregulated IL-6 and TNF-α in peripheral blood and increased IL-6 and iNOS in ischemia tissues were confirmed. The supernatant expression of TNF-α and IL-6, as well as IL-6, iNOS and CD86 mRNA, was significantly increased in the of Bv-2 cells after oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in vitro. For further understanding the expression pattern of RNAs, the next-generation RNA sequencing was performed and upregulation of Robo4 (roundabout guidance receptor 4) was found both in M1-polarized and OGD/R treated Bv-2 cells, which was also confirmed by RT-qPCR. Extracellular soluble Robo4 (sRobo4) protein also increased in the supernatant of M1-polarized and OGD/R treated Bv-2 cells. Treating bEND3 cells with the Robo4 recombinant protein, M1-polarized Bv-2 cell supernatant or OGD/R Bv-2 cell supernatant decreased trans-endothelial electrical resistance (TEER), suggesting the injury of BBB. In addition, Robo4 was also highly expressed in the serum of patients who experienced acute ischemia stroke and mechanical thrombectomy operation. All the results suggest that increased secretion of Robo4 by M1-polarized-microglia during cerebral ischemia-reperfusion is most likely one of the causes of BBB injury, and Robo4 may be one of the therapeutic targets for BBB functional protection.

Fasudil on Myocardial Injury in Rats with Myocardial Ischemia and Reperfusion through Rho-ROCK Signal Pathway.

Cardiovascular diseases are very harmful to human life and health. Reperfusion therapy is a standard method to treat cardiovascular diseases and has achieved high clinical effects. However, this treatment method is likely to cause myocardial ischemia-reperfusion injury. It has been reported that the Rho kinase inhibitor fasudil can interfere with cardiomyocyte apoptosis through the Rho-ROCK signaling pathway, so it is often used to treat cardiovascular diseases. The essay aims to research this specific influence of fasudil on cardiac damage in myocardial ischemia-reperfusion mouses through the Rho-ROCK signal path and its related mechanisms. Forty rats were taken as the research object, and the mouses were separated into control clusters. In the observation cluster of fasudil, the rat heart device was perfused by surgery. The rat coronary artery was ligated for 20 minutes to make the rat myocardial ischemia. Then, the ligation was loosened for myocardial perfusion to create a rat myocardial ischemia-reperfusion model. Observation group rats were perfused with quantitative fasudil, 80 minutes after ischemia-reperfusion, the ultrastructural changes and myocardial ischemic area of the rat myocardium were observed under a microscope, and the dynamic changes of the mouse heart were examined by flow cytometry. The PCR fluorescence method was used to explore the outlook layer of Rho-ROCK kinase activity to detect rat cardiomyocyte apoptosis. It is shown that under this intervention of fasudil, this expression level of Rho-ROCK kinase activity in the observation group was reduced by 18.3%, the myocardial cell apoptosis rate was decreased by 26.4%, and one area of myocardial ischemia can be reduced by 32.5%. The ultrastructure of the new object in rats is improved, and the left ventricular diastolic and systolic effect is enhanced. Therefore, the fasudil may decrease cardiac ischemia and focus on injured Rho-ROCK signal path activity.

Epigenetic silencing of E-cadherin gene induced by lncRNA MALAT-1 in acute myeloid leukaemia.

BACKGROUND: Epigenetic abnormalities in acute myeloid leukaemia provide us with a target for novel therapeutic strategies. The aim of the study was to verify the epigenetic regulatory mechanism of E-cadherin gene silencing induced by long non-coding RNA MALAT-1 in AML. METHODS: Expression of MALAT-1, E-cadherin, EZH2, SUZ12 and EED genes in AML patients was detected by RT-qPCR. After MALAT-1 silencing in AML cell lines, levels of the E-cadherin, EZH2, SUZ12, EED, DNMT1, DNMT3A and DNMT3B genes and encoded proteins were detected by RT-qPCR and Western blotting. The level of CpG island methylation and trimethylation modification of histone H3K27 in the promoter region of E-cadherin was detected by pyrosequencing and ChIP-qPCR. RIP-qPCR was used to detect the interaction between MALAT-1 and proteins. RESULTS: MALAT-1, EZH2 and EED gene expression was markedly increased in AML patients with E-cadherin down-regulation. A positive correlation between EZH2 or SUZ12 and MALAT-1 expression was observed. After MALAT-1 silencing, the expression of E-cadherin was up-regulated, whereas the expression of EZH2, SUZ12, DNMT1, DNMT3A and DNMT3B was down-regulated. Results of Western blotting were consistent with those of RT-qPCR. Methylation levels of E-cadherin in AML patients were higher than that in normal controls, which appeared to increase with age. Methylation of the CpG island and H3K27 trimethylation of E-cadherin were decreased after MALAT-1 silencing. RIP-qPCR suggested that MALAT-1 might be enriched by EZH2 and SUZ12. CONCLUSION: Our findings verified that MALAT-1 might lead to the transcriptional silencing of E-cadherin gene through the trimethylation of H3K27 mediated by recruiting EZH2 and SUZ12.

Chromobox Homologue 7 Acts as a Tumor Suppressor in Both Lung Adenocarcinoma and Lung Squamous Cell Carcinoma via Inhibiting ERK/MAPK Signaling Pathway.

Chromobox homologue 7 (CBX7) is a member of the polycomb group family that plays a pivotal role in regulating cellular processes in human cancers. This study aims to explore the function and underlying molecular mechanisms of CBX7 in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). The expression of CBX7 in LUAD and LUSC tissues was analyzed by UALCAN and GEPIA based on the TCGA database. Cell viability and apoptosis were measured by CCK-8 and flow cytometry assays, respectively. Cell migration and invasion were detected by transwell assay. The functions of downregulated genes in LUAD were enriched via GO and KEGG pathway analyses. The mRNA expression of CBX7, ERK1/2, and p38 was determined by qRT-PCR, and the protein levels of CBX7, ERK1/2, p-ERK1/2, p38, and p-p38 were measured by Western blotting. Tumor xenograft model was established to validate the antitumor effect of CBX7. The expression of CBX7 and Ki-67 in tumor tissues was detected by immunohistochemistry. CBX7 was downregulated in the tissues and cells of both LUAD and LUSC. Low CBX7 expression was associated with a poor overall survival rate in LUAD patients. CBX7 overexpression inhibited the viability, migration, and invasion and promoted the apoptosis of LUAD and LUSC cells. In addition, the downregulated genes in LUAD were enriched in MAPK cascade (GO) and MAPK signaling pathway (KEGG). ERK/MAPK pathway was then determined as a downstream target of CBX7, which was inhibited by CBX7 overexpression in LUAD and LUSC cells. The overexpression of CBX7 inhibited the malignant progression of LUAD and LUSC cells probably via suppressing the ERK/MAPK signaling pathway in vitro and in vivo.