Determination of the 95% effective dose of remimazolam tosylate in anesthesia induction inhibits endotracheal intubation response in senile patients.

Background and Purpose: The prevalence of elderly patients prompts anesthesiologists to determine the optimal dose of medication due to the altered pharmacokinetics and pharmacodynamics of this population. The present study aimed to determine the 95% effective dose (ED95) of remimazolam tosylate in anesthesia induction to inhibit endotracheal intubation-related cardiovascular reaction in frail and non-frail senile patients. Methods: A prospective sequential allocation dose-finding study of remimazolam tosylate was conducted on 80 elderly patients who received general anesthesia between May and June 2022 at the First Affiliated Hospital of Nanchang University. The initial dose was 0.3 mg/kg. The blood pressure and heart rate fluctuations during intubation were either <20% (negative cardiovascular response) or ≥20% (positive cardiovascular response). If positive, the dose of the next patient was increased by 0.02 mg/kg, while if negative, it was reduced by 0.02 mg/kg according to the 95:5 biased coin design (BCD). The ED95 and 95% confidence intervals (CIs) were determined using R-Foundation isotonic regression and bootstrapping methods. Results: The ED95 of remimazolam tosylate to inhibit the response during tracheal intubation was 0.297 mg/kg (95% CI: 0.231-0.451 mg/kg) and 0.331 mg/kg (95% CI: 0.272-0.472 mg/kg) in frail and non-frail senile patients, respectively. Conculation and Implications: The CI of the two groups overlap, and no difference was detected in the ED95 of remimazolam tosylate in inhibiting endotracheal intubation-related cardiovascular response in frail and non-frail senile patients. These results suggested that remimazolam tosylate is an optimal anesthesia inducer for all elderly patients. Clinical Trial Registration: https://www.chictr.org.cn, identifier ChiCTR2200055709.

Low-intensity transcutaneous auricular vagus nerve stimulation reduces postoperative ileus after laparoscopic radical resection of colorectal cancer: a randomized controlled trial.

BACKGROUND: Postoperative ileus (POI) is thought to result from a disrupted sympathetic/parasympathetic balance caused by trauma or surgery. Transcutaneous auricular vagus nerve stimulation (tVNS) is a non-invasive technique involving stimulation of the vagal auricular branch, leading to autonomic regulation and reduced inflammation. Here, the effects of low-intensity transcutaneous auricular vagal stimulation on POI after laparoscopic radical resection of colorectal cancer were investigated. METHODS: One hundred and thirty-four patients who received scheduled laparoscopic radical resection of colorectal cancer were randomly allocated to the A and B groups. The A group received low-intensity (25 Hz, 50 mA) transcutaneous electrical stimulation of the right auricular branch for 20 minutes prior to anesthesia while the B group did not. The primary outcome was the incidence of POI. RESULTS: The incidence of POI in the A group was 6.25% and 20% in the B group (P=0.022). Patients in the A group showed more regular bowel sounds after 24, 36, and 48 h than those in the B group (P<0.001). CONCLUSIONS: Low-intensity transcutaneous auricular vagal stimulation reduced POI after laparoscopic radical resection of colorectal cancer.

Lidocaine inhibits the proliferation and migration of endometrial cancer cells, and promotes apoptosis by inducing autophagy.

As a gynecological malignancy, endometrial cancer (EC) has a high incidence and mortality rate in women. The aim of the present study was to investigate the mechanism of EC and to identify novel effective treatment methods for this disease. The viability and proliferation of the RL95-2 human endometrial cancer cell line were assessed using Cell Counting Kit-8 assays. Colony formation, wound healing, Transwell, TUNEL and immunofluorescence assays were used to assess the effects of 5, 10 and 15 mM lidocaine on the colony formation, migration, invasiveness, apoptosis and Beclin 1 protein expression of RL95-2 cells, respectively. Furthermore, western blotting was used to analyze the protein expression levels of apoptosis- and autophagy-related proteins. The results demonstrated that lidocaine inhibited the viability, proliferation and migration of EC cells, and promoted apoptosis. Furthermore, lidocaine was demonstrated to induce autophagy and Beclin 1 protein expression in EC cells. In conclusion, lidocaine inhibited the proliferation and migration of EC cells, and promoted apoptosis via autophagy induction, which indicated that lidocaine may be a potential therapeutic drug for the treatment of EC.

Lidocaine promotes apoptosis in breast cancer cells by affecting VDAC1 expression.

OBJECTIVE: To investigate the effect of lidocaine on the expression of voltage-dependent anion channel 1 (VDAC1) in breast invasive carcinoma (BRCA) and its impact on the apoptosis of breast cancer cells. METHODS: We collected clinical data from patients with invasive breast cancer from 2010 to 2020 in the First affiliated hospital of Nanchang University, evaluated the prognostic value of VDAC1 gene expression in breast cancer, and detected the expression of VDAC1 protein in breast cancer tissues and paracancerous tissues by immunohistochemical staining of paraffin sections. Also, we cultured breast cancer cells (MCF-7) to observe the effect of lidocaine on the apoptosis of MCF-7 cells. RESULTS: Analysis of clinical data and gene expression data of BRCA patients showed VDAC1 was a differentially expressed gene in BRCA, VDAC1 may be of great significance for the diagnosis and prognosis of BRCA patients. Administration of lidocaine 3 mM significantly decreased VDAC1 expression, the expression of protein Bcl-2 was significantly decreased (p < 0.05), and the expression of p53 increased significantly (p < 0.05). Lidocaine inhibited the proliferation of MCF-7 breast cancer cells, increased the percentage of G2 / M phase cells and apoptosis. CONCLUSION: Lidocaine may inhibit the activity of breast cancer cells by inhibiting the expression of VDAC1, increasing the apoptosis in breast cancer cells.

Acidic preconditioning reduces lipopolysaccharide-induced acute lung injury by upregulating the expression of angiotensin-converting enzyme 2.

Acid preconditioning (APC) through carbon dioxide inhalation can exert protective effects during acute lung injury (ALI) triggered by ischemia-reperfusion. Angiotensin-converting enzyme 2 (ACE2) has been identified as a receptor for severe acute respiratory syndrome coronavirus and the novel coronavirus disease-19. Downregulation of ACE2 plays an important role in the pathogenesis of severe lung failure after viral or bacterial infections. The aim of the present study was to examine the anti-inflammatory mechanism through which APC alleviates lipopolysaccharide (LPS)-induced ALI in vivo and in vitro. The present results demonstrated that LPS significantly downregulated the expression of ACE2, while APC significantly reduced LPS-induced ALI and provided beneficial effects. In addition, bioinformatics analysis indicated that microRNA (miR)-200c-3p directly targeted the 3'untranslated region of ACE2 and regulated the expression of ACE2 protein. LPS exposure inhibited the expression of ACE2 protein in A549 cells by upregulating the levels of miR-200c-3p. However, APC inhibited the upregulation of miR-200c-3p induced by LPS, as well as the downregulation of ACE2 protein, through the NF-κB pathway. In conclusion, although LPS can inhibit the expression of ACE2 by upregulating the levels of miR-200c-3p through the NF-κB pathway, APC inhibited this effect, thus reducing inflammation during LPS-induced ALI.

[Effects of stachyine on apoptosis in an Aß25-35-induced PC12 cell model of Alzheimer's disease].

OBJECTIVE: To investigate the effects of stachydrine (STA) on apoptosis of Aß25-35-induced PC12 cells mimicking Alzheimer's disease and explore the mechanisms. METHODS: The differential genes of STA were analyzed based on GSE85871 data, and the target genes of STA were identified using STITCH database. PC12 cells were treated with Aß25-35 to establish a cell model of Alzheimer's disease, and the changes in cell viability and cell cycle in response to STA treatment were assessed using MTT assay and flow cytometry, respectively. RT-PCR and Western blotting were used to detect the relevant gene or protein expressions in the treated cells. RESULTS: GSE85871 data showed 37 up-regulated genes and 48 down-regulated genes in cells following treatment with STA. Analysis of the data from the STITCH database indicated that RPS8 and EED were the target genes of STA. Treatment of PC12 cells with Aß25-35 significantly lowered the cell viability (P < 0.05) and the expressions of RPS8 and EED at both the mRNA and protein levels (P < 0.05), and obviously inhibited the expression of apoptosis-related proteins Bcl-2 and p53 (P < 0.05). STA treatment of the cells significantly reversed the effect of Aß25-35 and induced cell cycle arrest in G2/M phase, causing also significantly increases in the expression levels of RPS8, EED, Bcl-2 and p53 (P < 0.05). CONCLUSIONS: STA plays an important role in inhibiting the apoptosis of PC12 cells induced by Aß25-35 possibly by regulating RPS8 and EED expression to promote the expressions of Bcl-2 and p53.

Propofol upregulates miR-320a and reduces HMGB1 by downregulating ANRIL to inhibit PTC cell malignant behaviors.

BACKGROUND: Our previous study states that propofol suppresses proliferation and migration of papillary thyroid cancer (PTC) cells by downregulation of lncRNA ANRIL. This study intended to probe the downstream mechanism of ANRIL in PTC with potential microRNAs (miR) and genes. METHODS: ANRIL expression was detected in normal thyroid epithelial cells (Nthy-ori 3-1) and PTC cells (TPC-1, FTC-133, K1 and BCPAP). ANRIL expression was inhibited in TPC-1 and BCPAP cells to explore the effects of si-ANRIL in PTC malignant behaviors. The gain-and loss-of functions of ANRIL/miR-320a were performed to measure their roles in PTC. Levels of ANRIL, miR-320a, HMGB1, apoptosis- and Wnt/ß-catenin and NF-κB pathways-related proteins were measured. Dual-luciferase reporter gene assay and RNA pull-down assay were applied to verify ANRIL/miR-320a/HMGB1 relation. si-ANRIL was transplanted into xenograft tumors in nude mice. RESULTS: ANRIL was upregulated in TPC-1 and BCPAP cells. miR-320a targeted HMGB1, and ANRIL bound to miR-320a. In TPC-1 and BCPAP cells, si-ANRIL prevented PTC cell malignant behaviors, and inactivated the Wnt/ß-catenin and NF-κB pathways; while si-ANRIL + miR-320a inhibition showed opposite trends. Overexpressing miR-320a promoted malignant behaviors of TPC-1 cells. In 6 µg/mL propofol-treated TPC-1 cells, miR-320a inhibition weakened propofol's inhibitory effects on PTC cell growth. After ANRIL inhibition, the volume and weight of xenograft tumors were decreased. CONCLUSION: Propofol upregulated miR-320a and reduced HMGB1 by downregulating ANRIL and inactivating the Wnt/ß-catenin and NF-κB pathways, thus preventing PTC cell malignant behaviors. This study may offer new insights in PTC prevention and treatment.

Acidic Preconditioning Protects Against Ischemia-Reperfusion Lung Injury Via Inhibiting the Expression of Matrix Metalloproteinase 9.

BACKGROUND: Acidic preconditioning (APC) has been demonstrated to protect against ischemia-reperfusion (IR)-induced lung injury, which could occur during lung transplantation or cardiopulmonary bypass. However, the pathophysiological mechanisms underlying IR lung injury and APC protection are not completely understood. The key factors responsible for the protective effects of APC are not clear. In this study, bioinformatics was used to predict the potential key factor in IR lung injury and explore the important mediator of the APC protective effect in IR lung injury. METHODS: First, we screened GSE6730, which is related to both lung injury and IR in Gene Expression Omnibus, and STRING was used later to select the genes in GSE6730 needed in the future. Animal models were established and classified to validate the effect of matrix metalloproteinase 9 (MMP-9) on lung injury after IR by adding a selective inhibitor (4-phenoxyphenylsulfonyl) methylthiirane, MMP-9 inhibitor. Next, for better understanding of APC inhibition of the expression of MMP-9 in lung injury, assessment of lung tissues, Western blot analysis, and RNA extraction and reverse transcription quantitative polymerase chain reaction were conducted. RESULTS: MMP-9 was identified to be overexpressed after IR according to the analysis on GSE67370. MMP-9 was an unknown gene in relation to acute lung injury and found to be associated with interleukin (IL)-1B, IL-6, and IL-8. The expressions of these inflammatory factors, including MMP-9, were all elevated in IR. Furthermore, lung injury was ameliorated, and the level of MMP-9 was lower when an MMP-9 inhibitor, (4-phenoxyphenylsulfonyl) methylthiirane, was added. Compared with group IR, APC reversed the ischemia-induced lung injury, and the level of MMP-9 was lower, and the concentrations of IL-1ß, IL-6, and IL-8 were decreased. CONCLUSIONS: Our findings reveal a novel mechanism indicating that IR induces higher expression of MMP-9 in lung injury by increasing the expression of inflammation-related factors. APC might protect against IR lung injury by inhibiting the expression of MMP-9.

[Low-intensity pulsed ultrasound pretreatment inhibits HMGB1 expression and attenuates lung ischemia-reperfusion injury in rats via the cholinergic anti-inflammatory pathway].

OBJECTIVE: To observe the effects of low-intensity pulsed ultrasound (LIPUS) pretreatment on pulmonary expression of high mobility group box-1 (HMGB1) in a rat model of lung ischemia-reperfusion (IR). METHODS: Thirty-two male SpragueDawley rats weighing 250-300 g were randomly divided (n=8) into sham-operated group, lung IR group, LIPUS pretreatment group and pretreatment with α7-nicotinic cholinergic receptor (α7nAChR) antagonist group. In the sham-operated group, the left pulmonary hilum was dissociated without occlusion; in the other 3 groups, the left pulmonary hilum was occluded for 45 min followed by reperfusion for 180 min; LIPUS pretreatment for 30 min and intraperitoneal injection of methyllycaconitine (2 mg/kg), an α7nAChR antagonist, were administered before the operation. The wet/dry weight ratio (W/D) and pulmonary permeability index (LPI) of the lung tissue were measured, and the lung histopathology was observed and scored. The contents of interleukin-1 (IL-1) and IL-6 in the lung tissues were measured using ELISA, and the pulmonary expression of HMGB1 protein was detected using immunofluorescence assay and Western blotting. RESULTS: Compared with those in the sham-operated group, the W/D of the lung tissue, LPI, pathological scores, IL-1 and IL-6 contents in the lung tissue, and pulmonary HMGB1 expression all significantly increased in the other 3 groups (P < 0.05). LIPUS preconditioning significantly lowered the W/D values, LPI, pathological score, IL-1 and IL-6 contents and HMGB1 expression in the lung tissues following lung IR, and these effects were significantly inhibited by administration of methyllycaconitine. CONCLUSIONS: LIPUS preconditioning can reduce lung IR injury possibly by activating α7nAChR-dependent cholinergic anti-inflammatory pathway to reduce lung tissue HMGB1 expression.

[Effects of acidification pretreatment for respiratory acidosis on the expression of matrix metalloproteinase-9 in rat lung tissues following ischemia/reperfusion].

OBJECTIVE: To establish rat model of lung ischemia/reperfusion (IR) in vivo, and to explore the effects of acidification pretreatment for respiratory acidosis on the expression of matrix metalloproteinase-9 (MMP-9) and the possible mechanisms.
 Methods: A total of 36 male Sprague-Dawley rats were divided into a sham group (S group), a IR group, and an experiment group (RA group) (n=12 in each group). The rat left lung hilum in the S group was dissociated, followed by perfusion without ischemia. After the left lung hilum in the IR group was blocked for 45 min, the rats were followed by reperfusion for 180 min. After left lung hilum in the RA group was dissociated, the respiratory parameters were adjusted so that pressure of end tidal carbon dioxide (PETCO2) reached 56-65 mmHg (1 mmHg=0.133 kPa) for 5 min, then the rats was subjected to IR. Lung tissue wet/dry (W/D) and lung permeability index (LPI) were calculated, while the lung histopathology was observed and the MMP-9 protein expression were measured.
 Results: Compared with the control group, the W/D and LPI in the IR group and the RA group increased after reperfusion (both P<0.05), and the levels of W/D and LPI in the group RA were lower than that in the IR group (P<0.05). LPI and pathology scores were significantly lower in the RA group than those in the IR group (both P<0.01). After IR, the expression of MMP9 in the lung tissues in the IR group and the RA group increased significantly (both P<0.01). The expression of MMP-9 protein in the RA group was significantly lower than that in the IR group (P<0.01).
 Conclusion: After lung IR injury, the expression of MMP-9 protein, vascular permeability and inflammatory exudation is increased. The acidification pretreatment for respiratory acidosis can inhibit the expression of MMP-9 protein and reduce inflammatory exudation after lung IR, showing a protective effect on lung IR injury.