TXNIP knockdown protects rats against bupivacaine-induced spinal neurotoxicity via the inhibition of oxidative stress and apoptosis.

Bupivacaine (BUP) is an anesthetic commonly used in clinical practice that when used for spinal anesthesia, might exert neurotoxic effects. Thioredoxin-interacting protein (TXNIP) is a member of the α-arrestin protein superfamily that binds covalently to thioredoxin (TRX) to inhibit its function, leading to increased oxidative stress and activation of apoptosis. The role of TXNIP in BUP-induced oxidative stress and apoptosis remains to be elucidated. In this context, the present study aimed to explore the effects of TXNIP knockdown on BUP-induced oxidative stress and apoptosis in the spinal cord of rats and in PC12 cells through the transfection of adeno-associated virus-TXNIP short hairpin RNA (AAV-TXNIP shRNA) and siRNA-TXNIP, respectively. In vivo, a rat model of spinal neurotoxicity was established by intrathecally injecting rats with BUP. The BUP + TXNIP shRNA and the BUP + Control shRNA groups of rats were injected with an AAV carrying the TXNIP shRNA and the Control shRNA, respectively, into the subarachnoid space four weeks prior to BUP treatment. The Basso, Beattie & Bresnahan (BBB) locomotor rating score, % MPE of TFL, H&E staining, and Nissl staining analyses were conducted. In vitro, 0.8 mM BUP was determined by CCK-8 assay to establish a cytotoxicity model in PC12 cells. Transfection with siRNA-TXNIP was carried out to suppress TXNIP expression prior to exposing PC12 cells to BUP. The results revealed that BUP effectively induced neurological behavioral dysfunction and neuronal damage and death in the spinal cord of the rats. Similarly, BUP triggered cytotoxicity and apoptosis in PC12 cells. In addition, treated with BUP both in vitro and in vivo exhibited upregulated TXNIP expression and increased oxidative stress and apoptosis. Interestingly, TXNIP knockdown in the spinal cord of rats through transfection of AAV-TXNIP shRNA exerted a protective effect against BUP-induced spinal neurotoxicity by ameliorating behavioral and histological outcomes and promoting the survival of spinal cord neurons. Similarly, transfection with siRNA-TXNIP mitigated BUP-induced cytotoxicity in PC12 cells. In addition, TXNIP knockdown mitigated the upregulation of ROS, MDA, Bax, and cleaved caspase-3 and restored the downregulation of GSH, SOD, CAT, GPX4, and Bcl2 induced upon BUP exposure. These findings suggested that TXNIP knockdown protected against BUP-induced spinal neurotoxicity by suppressing oxidative stress and apoptosis. In summary, TXNIP could be a central signaling hub that positively regulates oxidative stress and apoptosis during neuronal damage, which renders TXNIP a promising target for treatment strategies against BUP-induced spinal neurotoxicity.

A dynamic study of the postoperative management of thyroid cancer from 2003 to 2022: a bibliometric analysis.

Background: Over the past 20 years, the global incidence of thyroid cancer has continued to increase. The volume of literature on the postoperative management of thyroid cancer comprises 1,040 articles, from 64 countries, with 1,400 journals publishing the relevant literature, and several guidelines on the treatment of thyroid cancer. This study used bibliometric methods to identify research hotspots and explore future directions in this field. Methods: We comprehensively searched the Science Citation Index Expanded (SCI-E) database of the Web of Science Core Collection (WOSCC) for articles published from 2003 to 2022 on the postoperative management of thyroid cancer. Using CiteSpace 6.1.R6 and Microsoft Office Excel 2010, we evaluated and visualized the search results. Using R Studio, we generated a network of spatial geographic distribution maps and cooperative network. Results: A total of 1,040 publications were included in the study. The results revealed an overall upward trend in the number of publications and citations over the past 20 years. The United States of America (USA) had the largest number of publications and the highest centrality (n=282, centrality =0.28). Johns Hopkins University had highest centrality (centrality =0.15) and was the academic center of the field. Thyroid was the journal with the highest number of citations (n=826), and the American Journal of Surgical Pathology was the journal with the highest centrality (centrality =0.08). The top 10 citations in the literature were mainly guidelines and consensus statements on the management of thyroid cancer. A keyword-based clustering analysis revealed the prominence of clusters of keywords, such as follow-up, recurrent laryngeal nerve, and medullary thyroid carcinoma (MTC). A keyword burst detection analysis showed that the term papillary had the highest burst intensity (strength =8.02), while management guidelines, association guidelines, active surveillance (AS), microcarcinoma, and differentiated thyroid cancer were the current burst words. Conclusions: Over the past two decades, the number of relevant publications in the postoperative management of thyroid cancer field has continued to grow. Among the many research directions, follow-up, recurrent laryngeal nerve, and MTC are research hotspots. Future research is likely to revolve around guidelines and consensus statements on the management of thyroid cancer, AS, and microcarcinoma in differentiated thyroid cancer.

Construction and validation of a clinical prediction model for asymptomatic obstructive coronary stenosis in patients with carotid stenosis.

Background: Coronary artery stenosis occurs frequently in patients with carotid artery stenosis. We developed a clinical predictive model to investigate the clinical risk of asymptomatic obstructive coronary artery stenosis in patients with carotid artery stenosis ≥ 50%. Methods: From January 2018 to January 2022, carotid stenosis patients hospitalized at the First Affiliated Hospital of Zhengzhou University's Department of Endovascular Surgery were subjected to a retrospective analysis of their clinical information and imaging results. Excluded criteria were patients with lacking data, symptomatic coronary stenosis, prior coronary artery bypass grafting, and coronary stent implantation. Patients were separated into case and control groups according to whether or not they had obstructive coronary stenosis. Independent predictors were screened using univariate and multivariate logistic regression, and their accuracy was confirmed using least absolute shrinkage and selection operator (LASSO) regression. A Nomogram prediction model was developed using the aforementioned filtered factors. The model's discrimination and specificity were evaluated using the receiver operating characteristic curve (ROC) and Hosmer-Lemeshow goodness-of-fit test. Internal validation employed the Bootstrap procedure. The clinical decision curve analysis (DCA) of the prediction model was developed to assess the clinical applicability of the model. Results: The investigation included a total of 227 patients, of whom 132 (58.1%) had coronary artery stenosis. Hypertension, Grade I plaque, HbA1c ≥ 7.0%, MPV ≥ 9.2fl, and Fib ≥ 3.0 g/L were independent predictors, with OR values of (2.506, 0.219, 0.457, 1.876, 2.005), according to multivariate logistic regression. Risk factor screening and validation using lasso regression. The predictors chosen based on the optimal λ value are consistent with the predictors identified by multiple regression. The area under the ROC curve (AUC) of the model based on the above predictors was 0.701 (0.633-0.770), indicating that the model discriminated well. The calibration curve of the model closely matched the actual curve, and P > 0.05 in the Hosmer-Lemeshow goodness-of-fit test indicated the model's accuracy. The results of the DCA curve demonstrate the clinical applicability of the prediction model. Conclusion: Hypertension, grade I plaque, HbA1c ≥ 7.0%, MPV ≥ 9.2 fl, and Fib ≥ 3.0 g/L are predictors of asymptomatic coronary stenosis in patients with carotid stenosis ≥50%. The diagnostic model is clinically applicable and useful for identifying patients at high risk.

Ferrostatin-1 ameliorates Bupivacaine-Induced spinal neurotoxicity in rats by inhibiting ferroptosis.

Bupivacaine (BUP) has previously been shown to trigger neurotoxicity after spinal anesthesia. Further, ferroptosis has been implicated in the pathological processes associated with various central nervous system diseases. Although the impact of ferroptosis on BUP-induced neurotoxicity in the spinal cord has not been fully understood, this research aims to investigate this relationship in rats. Additionally, this study aims to determine whether ferrostatin-1 (Fer-1), a potent inhibitor of ferroptosis, can provide protection against BUP-induced spinal neurotoxicity. The experimental model for BUP-induced spinal neurotoxicity involved the administration of 5% bupivacaine through intrathecal injection. Then, the rats were randomized into the Control, BUP, BUP + Fer-1, and Fer-1 groups. BBB scores, %MPE of TFL, and H&E and Nissl stainings showed that intrathecal Fer-1 administration improved functional recovery, histological outcomes, and neural survival in BUP-treated rats. Moreover, Fer-1 has been found to alleviate the BUP-induced alterations related to ferroptosis, such as mitochondrial shrinkage and disruption of cristae, while also reducing the levels of malondialdehyde (MDA), iron, and 4-hydroxynonenal (4HNE). Fer-1 also inhibits the accumulation of reactive oxygen species (ROS) and restores the normal levels of glutathione peroxidase 4 (GPX4), cystine/glutamate transporter (xCT), and glutathione (GSH). Furthermore, double-immunofluorescence staining revealed that GPX4 is primarily localized in the neurons instead of microglia or astroglia in the spinal cord. In summary, we demonstrated that ferroptosis play a pivotal role in mediating BUP-induced spinal neurotoxicity, and Fer-1 ameliorated BUP-induced spinal neurotoxicity by reversing the underlying ferroptosis-related changes in rats.

Enhanced induction of heme oxygenase-1 suppresses thrombus formation and affects the protein C system in sepsis.

Heme oxygenase-1 (HO-1) displays anti-inflammatory and cytoprotective activities in sepsis. Here, we investigated the effects of HO-1 on thrombus formation and the protein C system in a septic C57BL/6 mouse model induced by cecal ligation and perforation (CLP). Septic mice were either preinjected with the vehicle, pretreated with hemin (an HO-1 inducer) or zinc protoporphyrin IX (ZnPP, an HO-1 inhibitor), or given a combination of hemin + ZnPP. CLP increased significantly the hepatic expression of HO-1; increased thrombosis in livers, kidneys, and lungs; shortened the prothrombin time (PT) and activated partial thromboplastin time (APTT); elevated the levels of tumor necrosis factor-1α (TNF-1α), interleukin-6 (IL-6), and thrombomodulin (TM); reduced the levels of protein C (PC) and activated protein C (aPC); and downregulated hepatic expression of PC and TM. The preadministration of hemin to septic mice increased the expression and activity of HO-1; inhibited thrombosis in the preceding 3 organs; prolonged PT and APTT; inhibited the production of TNF-α and IL-6; upregulated the expression of PC and TM in livers; elevated the plasma levels of PC and aPC; and reduced the plasma levels of TM. In contrast, ZnPP showed opposite effects to hemin and reversed the effects of hemin by inhibiting the activity of HO-1. The administration of tricarbonyl dichloro ruthenium (II) dimer (CORM-2), which is a CO-releasing molecule, had a similar effect to hemin on thrombosis and the protein C system. The data indicate that the enhanced induction of HO-1 inhibits thrombus formation and affects the protein C system in sepsis.

[Artesunate suppresses human endometrial carcinoma RL95-2 cell proliferation by inducing cell apoptosis].

OBJECTIVE: To investigate the inhibitory effect of artesunate on human endometrial carcinoma RL95-2 cell line proliferation in vitro and the possible mechanisms. METHODS: The inhibitory effect of artesunate on the cell proliferation was assessed with MTT assay. Transmission electron miscrosopy was used to observe the morphological change of the cells after the treatment. Flow cytometry was performed to examine the changes in the cell cycle, reactive oxygen species (ROS) levels (with DCFH-DA labeling) and mitochondrial membrane potential (rhodamine123 staining), and caspase-3 activity was detected by immunohistochemistry. RESULTS: Artesunate inhibited the proliferation of RL95-2 cells with an IC(50) of 26.29 microg/ml. Transmission electron microscopy revealed early apoptotic changes of the cells with obvious chromatin fragmentation. The cell cycle arrest at G(0)/G(1) phase was observed by flow cytometry, and immunohistochemistry demonstrated caspase-3 positivity in cytoplasm. ROS generation in the cells increased obviously after treatment with artesunate for 72 h, which also resulted in lowered mitochondrial membrane potential. CONCLUSION: Artesunate suppressed the proliferation of RL95-2 cells in vitro possibly by inducing cell apoptosis.

[Cell cycle block and apoptosis in rabbit lens epithelial cells induced by hydroxycamptothecine].

OBJECTIVE: To study the cell cycle block and apoptosis induced by hydroxycamptothecine (HCPT) in cultured rabbit lens epithelial cells (RLECs) and its affect on the gene expression of fas/fas-L in RLECs. METHODS: Flow cytometry was used to examine HCPT-induced cell cycle block in cultured RLECs. We observed cell morphological changes with light microscope and electron microscope. Apoptosis was detected by terminal deoxyribose transferase-mediated deoxy-uridine triphosphate nick end labeling (TUNEL) assay. The TUNEL assay was performed following the manufacturer's instruction of in situ cell death detection. Expression of Fas and Fas-L was estimated by immunohistochemical staining. RESULTS: The flow cytometric analysis showed that the HCPT induced cell cycle block was at S phase. The ratio of nucleus and cytoplasm was increased. The electron microscopic examinations demonstrated that the HCPT initiated apoptosis with typical morphological features. The apoptosis of RLECs treated with HCPT was observed by TUNEL assay, and the number of the cells with apoptosis was increased with the increase of the HCPT concentration. The HCPT did not affect the gene expression of Fas/FasL. CONCLUSIONS: HCPT not only induces mitotic block but also induces apoptosis of RLECs which is not mediated by the Fas/FasL signaling pathway.

Interaction between cisplatin, 5-fluorouracil and vincristine on human hepatoma cell line (7721).

AIM:To evaluate the killing effects of CDDP, 5-Fu and VCR on human hepaoma cell line (7721).METHODS:The median-effect principle was used.RESULTS:Killing effects of the individual drug were enhanced as the median concentration increased. Antagonism was produced when two drugs were used at a higher concentration (CI > 1), and synergism was achiened when CI < 1. Finally, the effect was influenced by both the ratios of drug concentration and the sequence of administration.CONCLUSION:The drug administration order and drug concentrations are significant factors that need to be considered in clinical practice.