AIM: To investigate the effect of Astragaloside Ⅳ (As-Ⅳ) on low-glucose mediated tumor immunosuppression microenvironment and its molecular mechanism. METHODS: MTT assay was used to detect the effect of As-Ⅳ on the proliferation of CD4+T cells in low-glucose microenvironment in vitro. By ELISA experiment and qPCR detection of interleukin 2 (IL-2), interferon - gamma (IFN-γ), CD40L and transforming growth factor beta 1 (TGF-β1) level; Western blot was used to detect the expression of glucose transporter 1 (Glut-1), key glycolytic enzymes (HK, PFK1 and PK), AKT/mTOR signaling pathway and AKT/GSK3β signaling pathway in CD4+T cells. Molecular docking and AKT inhibitor experiments were used to verify the results. B16-PKM2-OE was used to establish a low-glucose tumor microenvironment animal model for verification. RESULTS: MTT assay showed that As-Ⅳ promoted the proliferation of CD4+T cells in low-glucose microenvironment (P<0.05). The results of ELISA and qPCR experiments showed that As-Ⅳ could increase the levels of IL-2, IFN-γ and CD40L, and reduce the level of TGF-β1 in tumor tissues (P<0.05). Western blot results showed that As-Ⅳ promoted Glut-1 protein expression on the surface of CD4+T cells, up-regulated the expression of glycolysis key enzymes, and activated AKT/mTOR and AKT/GSK-3β signaling in a concentration-dependent manner. Molecular docking and join AKT inhibitors As the experiment results indicate-Ⅳ activated AKT/mTOR signaling and AKT/GSK-3β signal; Animal experiments showed that As-Ⅳ exerted anti-tumor effect by activating the proliferation and activation of CD4+T cells in low-glucose microenvironment. CONCLUSION: As-Ⅳ promote sugar by activation of AKT/Glut signal micro environment of CD4+T cell proliferation and activation play a role of anti-tumor.

AIM: To elucidate the effects of cefoperazone-sulbactam on the pharmacokinetics of warfarin and the mechanism behind the enhancement of warfarin's efficacy. METHODS: Thirty-two rats were randomly assigned to four groups: WN (healthy rats after the gastric-administration of 0.125 mg/kg warfarin), WCN (healthy rats after the gastric-administration of 0.125 mg/kg warfarin and 0.3 g/kg cefoperazone-sulbactam), WO (a rat model of biliary drainage after the gastric-administration of 0.125 mg/kg warfarin), WCO (a rat model of biliary drainage after the gastric-administration of 0.125 mg/kg warfarin and 0.3 g/kg cefoperazone-sulbactam). Blood samples were collected at various time points from the femoral artery to determine the plasma concentration of warfarin and from the tail vein to measure the International Normalized Ratio (INR). Warfarin levels were quantified using LC/MS/MS, and pharmacokinetic parameters were calculated using a non-compartmental model. The expression of P-glycoprotein (P-gp) in the ileal tissues of the WN and WCN groups was determined by Western blotting. RESULTS: Compared with the WN group, the WCN group demonstrated a significant increase in AUC0-t, Cmax, and Ka, and a notable decrease in CL/F, and INR values significantly increased. However, there was no significant difference in pharmacokinetic parameters and INR values between the WO group and the WCO group. Compared to the WN group, the WO group showed a significant reduction in AUC0-t, Cmax, and CL/F, with an obvious increase in t1/2. The INR of the WCN group was significantly higher than that of the WCO group after 6 hours.Western blotting results indicated a 62.1% reduction in P-gp expression in the ileum of the WCN group compared to the WN group (P<0.01). CONCLUSION: Cefoperazone-sulbactam significantly influences the pharmacokinetic parameters of warfarin and enhances its pharmacological effect by inhibiting intestinal P-gp expression. Biliary drainage significantly affects the pharmacokinetics of warfarin rats, and there is no significant drug interaction between the two after biliary drainage.

AIM: To investigate the inhibitory effect and mechanism of hesperetin (HST) on human gefitinib-resistant NCI-H1975 lung adenocarcinoma cells. METHODS: CCK-8 assay was used to detect the effects of HST on the proliferation of NCI-H1975 cells; Annexin V-FITC/PI double staining was used to detect HST-induced apoptosis of NCI-H1975 cells; flow cytometry was used to observe the effects of HST and HST + acetylcysteine (NAC) combined on the levels of reactive oxygen species (ROS) in NCI-H1975 cells; Western blot was used to detect the expression of Bcl-2, Bax, Cleaved Caspase-3, p-eIF2α, eIF2α and CHOP proteins in NCI-H1975 cells by HST, NAC + HST and Salubrinal + HST. The antitumor effect of HST in vivo was studied by constructing a xenograft model in nude mice; HE staining was used to observe the effect of HST on the histopathological morphology of heart, liver, kidney and xenograft in tumor-bearing mice; immunohistochemistry was used to detect the effect of HST on p-eIF2α protein in xenograft tissues. RESULTS: Compared with the control group, HST at 37.5 μmol/L for 24 h significantly inhibited the viability of NCI-H1975 cells (P<0.05), and NAC attenuated the inhibitory effect of HST; concentrations greater than 150 μmol/L increased intracellular ROS levels (P<0.05), induced apoptosis (P<0.05), and increased Caspase3 activity (P<0.01), and compared with HST 300 μmol/L group, ROS levels, apoptosis rate, and Caspase3 activity were significantly decreased in NAC + HST 300 μmol/L group (P<0.01); HST up-regulated Bax, Cleaved Caspase-3, CHOP, and p-eIF2α expression and down-regulated Bcl-2 expression in a concentration-dependent manner (P<0.01), and compared with HST 300 μmol/L group, Bax and Cleaved Caspase-3 expression was decreased and Bcl-2 expression was increased in Sal + HST 300 μmol/L group; p-eIF2α and CHOP expression were significantly down-regulated in the NAC + HST 300 μmol/L and Sal + HST 300 μmol/L groups (P<0.01). In vivo, experiments showed that HST could significantly inhibit the growth of transplanted tumors and up-regulate p-eIF2α protein expression (P<0.05), and had no significant adverse effects on the growth status, body weight and important organs (heart, liver and kidney) of nude mice. CONCLUSION: HST inhibits the proliferation of gefitinib-resistant NCI-H1975 lung adenocarcinoma cells in vitro and in vivo, and the mechanism may be related to HST mediating ER stress-induced apoptosis of NCI-H1975 cells through ROS. 

AIM: To explore the protective effect and mechanism of melittin against LPS induced acute lung injury. METHODS: Mice were intratracheally injected with LPS (5 mg/kg) to establish acute lung injury model. The lung coefficient of mice and the lung wet-dry mass ratio were determined. The lung tissue was stained with HE. The number of inflammatory cells and expression levels of related inflammatory factors in bronchoalveolar lavage fluid (BALF) were detected. The activities of serum SOD, MDA and GSH were detected. The expression of TLR4, NF-κB p65 and p-NF-κB p65 in lung tissues were determined by immunohistochemistry and Western blot. RESULTS: Melittin had a protective effect on LPS-induced ALI, as evidenced by significantly reduced lung coefficient, lung wet/dry mass ratio, the number of inflammatory cells and the expression level of inflammatory factors in BALF, and improved pulmonary pathology (P<0.05, P<0.01). Compared with model group, the serum MDA level of melittin group was significantly decreased (P<0.01), while the activities of SOD and GSH were significantly increased (P<0.05, P<0.01). In addition, melittin can significantly reduce the expression of TLR4, inhibit the phosphorylation of NF-κB p65, and reduce the inflammatory response (P<0.01). CONCLUSION: Melittin can significantly improve LPS-induced ALI, and the protective effect of melittin is mainly related to the inhibition of TLR4/NF-κB signaling pathway and the reduction of oxidative stress and inflammation.

AIM: To investigate the potential of resveratrol (Res) to alleviate cerebral ischemia-reperfusion injury (CIRI) by activating autophagy. METHODS: One hundred SD rats were purchased and subjected to middle cerebral artery occlusion/ reperfusion (MCAO/R) using the suture method. Rats with failed surgeries were excluded, and they were randomly divided into seven groups: Sham (S) group, Model (M) group, Resveratrol pretreatment (Res) group, 3-methyladenine (3-MA) group, Resveratrol + 3-methyladenine (Res + 3-MA) group, Z-YVAD group, and Res + Z-YVAD group, with 12 rats in each group. Neurological deficits and brain damage were assessed using Longa scoring, TTC staining, and measurement of infarct volume. Immunohistochemistry, immunofluorescence, and immunoblotting were employed to detect the expression of autophagy-related proteins Beclin-1, LC3-II, P62, and pyroptosis-related proteins NLRP3, caspase-1, GSDMD, IL-1β. RESULTS: Resveratrol pretreatment improved neurological deficits and reduced infarct volume in rats. It upregulated autophagy-related proteins Beclin-1, LC3-II, downregulated P62, and pyroptosis-related proteins NLRP3, caspase-1, GSDMD, IL-1β expression. The autophagy inhibitor 3-MA completely reversed the above effects of Res. After adding the pyroptosis inhibitor Z-YVAD, the changes in autophagy-related proteins Beclin-1, LC3-II, and P62 were not significant, while the expression of NLRP3, caspase-1, GSDMD, IL-1β significantly decreased. Neurological deficits worsened, and infarct volume increased in rats. CONCLUSION: Res attenuates cerebral ischemia-reperfusion injury by activating autophagy and inhibiting pyroptosis, possibly through the regulation of the P62. 

AIM: To investigate the preventive effect and potential mechanism of bicyclol on sepsis-induced fulminant hepatitis (FH). METHODS: The FH model was established by lipopolysaccharide (LPS)/D-galactose (D-Gal), and mice were orally administrated with bicyclol and the survival rate within 24 h was recorded. The activities of glutamate aminotransferase (AST), alanine aminotransferase (ALT), catalase (CAT) and superoxide dismutase (SOD), and the content of glutathione (GSH) and malondialdehyde (MDA) in liver tissue were measured by chemiluminescence. The histopathological changes of liver were examined by HE and TUNEL staining. The levels of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β) and interleukin 6 (IL-6) were measured using ELISA. The protein expression of nuclear transcription factor kappa B inhibitory protein α (IκBα), phosphorylated IκBα (p-IκBα), nuclear transcription factor kappa B (NF-κB), nuclear red blood cell 2 related factor 2 (Nrf2), NAD(P)H:quinone oxidoreductase 1 (NQO1), and heme oxygenase 1 (HO1) in liver tissue was examined by Western blotting. RESULTS: Pre-treatment with bicyclol improved survival ratio of FH mice, reduced ALT and AST activities, alleviated liver tissue lesions, and lowered Suzuki score. Meanwhile, the levels of TNF-α, IL-1β, IL-6 and MDA were reduced, the GSH level and CAT and SOD enzyme activities were increased, the protein expression of p-IκBα and nuclear NF-κB was down-regulated, and the protein levels of IκBα, nuclear Nrf2, NQO1 and HO1 were up-regulated. Moreover, post-treatment with Bicyclol also significantly reduced ALT and AST activities in FH mice. CONCLUSION: Bicyclol exhibited remarkable hepaprotective efects on FH caused by LPS/D-Gal, the potential mechanism underlying the anti-infammatory and antioxidative activities of Bicyclol might be associated with the suppression of NF-κB signaling pathway and the activation of Nrf2/NQO1/HO1 signaling pathway.

AIM: To explore the mechanism of Periplaneta americana extracts (PA-CII-3) on anti-prostate cancer. METHODS: Prostate cancer cells PC3 in logarithmic growth phase were divided into a control group and an experimental group. The PA-CII-3 solutions of 1 000, 1 200, 1 400, 1 600, 1 800, and 2 000 μg/mL were added successively to the PC3 cells in experimental group, for screening out the appropriate drug concentration and best time to intervene by CCK-8 assay. And then, PA-CII-3 solution at 1 400 μg/mL was continuously intervened in PC3 cells for 72 hours. The cell proliferation ability was detected by CCK-8 assay and plate colony formation assay, the cell migration ability was tested by scratch wound assay, and the mRNA and protein expressions of ZAG were detected by Real time qPCR and Western blot methods, respectively. RESULTS: After PA-CII-3 intervention, the proliferation and migration abilities of PC3 cells were significantly inhibited, and the mRNA and protein expression levels of ZAG in PC3 cells were promoted. CONCLUSION: PA-CII-3 can play the role of anti-prostate cancer by inhibiting the proliferation and migration of PC3 cells, and upregulating ZAG. 

AIM: To develop and validate an ELISA method for the determination of secukinumab (SEC) in human plasma and apply it in psoriasis patients. METHODS: A double-antibody sandwich ELISA was developed using anti-secukinumab antibody as the capture antibody and biotin-labeled anti-secukinumab antibody as the detection antibody. The method was systematically validated. Nineteen patients with Psoriasis treated with standard dose of SEC were included. In order to determine trough concentrations of SEC, steady-state blood samples were collected after 24 weeks of treatment. Psoriasis area and severity index score was used to evaluate the response. RESULTS: The SEC concentration showed a good concentration-response relationship within the range of 1.25 to 80.00 μg/mL.The intra-batch and inter-batch precision and accuracy were ≤ 15.00%, and there was no hook effect in the range of 1.25 to 1 000 μg/mL. The median trough concentrations of 19 patients was 33.56 μg/mL (IQR: 32.55-45.98 μg/mL) with an inter-individual variation of 52.00% for body weight adjusted concentration of SEC. The SEC concentrations were not significantly different between the active group and remission group (P=0.92). CONCLUSION: We developed and validated a method for the determination of SEC, which can be used for therapeutic drug monitoring in patients receiving SEC therapy. However the inter-individual variation is large. Further study is needed to explore the association of SEC levels with clinical response in Psoriasis.

AIM: To analyze and screen the risk factors for acute kidney injury (AKI) following heart transplantation (HT), and to establish a dynamic nomograms prediction model to forecast early AKI after HT. METHODS: A retrospective analysis was conducted on clinical data from HT recipients at Nanjing First Hospital from October 2012 to June 2024. Patients were divided into AKI and non-AKI groups based on AKI occurrence within 7 days post-surgery, with a 8:2 ratio for training and testing sets. Lasso regression and multivariable logistic regression were used to select influencing factors. A dynamic nomogram model was visualized using R. Internal validation was performed using 1 000 bootstrap samples. Model accuracy and discrimination were evaluated using the area under the receiver operating characteristic curve (AUC-ROC), calibration curves, and the Hosmer-Lemeshow goodness-of-fit test. The nomogram model was compared with the Cleveland score. RESULTS: The results of a multivariable logistic regression indicate that a history of atrial fibrillation (OR=9.647, 95% CI=1.961-47.470), vasoactive inotropic score (OR=1.094, 95% CI=1.012-1.183), intraoperative transfusion of red blood cells or plasma (OR=10.200, 95% CI=1.727-60.238), postoperative central venous pressure (OR=1.548, 95% CI=1.186-2.021), and postoperative use of vancomycin (OR=25.082, 95% CI=2.122-296.417) are independent risk factors for HT-AKI. The dynamic nomogram model achieved an AUC of 0.842 (95% CI: 0.676-0.971) in the test set, with a calibration plot showing a slope close to 1 and a Brier score of 0.173. The Hosmer-Lemeshow goodness-of-fit test (χ2=5.658, P=0.685) suggests good predictive performance of the model. Moreover, this model demonstrates superior discriminative ability compared to the Cleveland score. CONCLUSION: This study identified preoperative, intraoperative, and postoperative risk factors influencing the occurrence of HT-AKI. The developed dynamic nomogram model accurately identifies high-risk individuals for early HT-AKI and is convenient for clinical use.

AIM: To investigate the risk factors affecting serum digoxin concentration and the relationship between the level of the concentration and adverse reactions (ADR), and to provide a basis for the safe and rational use of digoxin. METHODS: The serum digoxin concentration (SDC) was monitored by enzyme amplification immunoassay technology. The basic data, laboratory indicators, concomitant medication and adverse reactions of patients were collected through the medical record system. RESULTS: Among the 55 patients, 12.73%, 76.36% and 10.91% of the patients were treated with < 0.125 mg/d, 0.125 mg/d and 0.25 mg/d digoxin, respectively. And the serum digoxin concentration increased in a dose dependent manner. Eleven patients (20%) developed ADR, the most common of which were arrhythmias. The SDC of patients with ADR was (2.55±1.59) ng/mL, which was significantly higher than that of the non-ADR group (0.93±0.63 ng/mL) (P<0.000 1). The SDC in the patients with increased serum creatinine was higher than that in the normal creatinine group, and patients with severe renal impairment demonstrated significantly higher concentration of digoxin than that in patients with normal renal function (P<0.05). The effect of concomitant administration of other drugs on the SDCof digoxin was not observed. CONCLUSION: The SDC varies greatly among individuals, and is associated with dosage and renal function. The higher the concentration of digoxin in serum, the higher the incidence of ADR. Therefore, monitoring the SDC of digoxin should be strengthened, so as to improve the level of individual treatment.

AIM: To predict voriconazole (VCZ)'s pharmacokinetics in patients with different degree of liver dysfunction. METHODS: The physiological based pharmacokinetics (PBPK) models were developed for VCZ based on drug's physico-chemical property and in-vitro data. Then liver dysfunction induced changes in physiological, anatomical parameter was integrated into the PBPK model. The clinical verified VCZ model was used to predict VCZ's PK in patients of different CYP2C19 genotype with different degree of liver dysfunction. RESULTS: The PBPK predicted the VCZ exposure in the patients with liver dysfunction with acceptable prediction error. CONCLUSION: The PBPK model developed here could evaluate the impact of liver dysfunction and CYP2C19 genotype on VCZ's pharmacokinetics and support individual dose adjustment of VCZ.

AIM: To evaluate the bioequivalence of the two rivaroxaban tablets in Chinese healthy subjects. METHODS: Twenty-eight subjects under fasting status and twenty-eight subjects under fed status were enrolled in the study. This study was designed as a four period, fully repetitive, crossover study. All subjects were administered test (T) and reference (R) rivaroxaban tablets (10 mg) under fasting and fed condition respectively. Liquid chromatography - tandem mass spectrometry was used to detect the concentrations of rivaroxaban in plasma. WinNonlin 7.0 was used to calculate the main pharmacokinetic parameters (PK) and to evaluate the bioequivalence. RESULTS: In fasting group, the main pharmacokinetic parameters of T and R preparation were as follows: Cmax were(186.57±56.41) and (187.61±50.89) ng/mL; AUC0-t were (1 156.21±335.85) and (1 177.59±343.72) h·ng·mL-1; AUC0-∞ were (1 235.77±384.03) and (1 223.53±392.10) ng·h·mL-1. The 90% confidential interval (CI) of the three main parameters were 90.81%-105.67%, 92.83%-103.85% and 95.04%-107.13%. The upper limit of the 90% CI for the test- to- reference ratio of the within-subject of Cmax, AUC0-t  and  AUC0-∞ were 1.56, 1.41 and 1.73. In fed group, the main pharmacokinetic parameters of T and R preparation were as follows: Cmax were (207.81±45.26) and (211.04±36.62) ng/mL; AUC0-t were (1 271.26±260.92) and (1 233.23±201.85) h·ng·mL-1; AUC0-∞ were (1 290.76±264.90) and (1 251.68±203.73) ng·h·mL-1. The 90% CI of the three main parameters were 92.82%-102.28%, 97.68%-106.68% and 97.71%-106.68%. The upper limit of the 90% CI for the test- to- reference ratio of the within-subject of Cmax, AUC0-t and AUC0-∞ were 1.76, 1.47 and 1.47. CONCLUSION:The two preparations of rivaroxaban tablets were bioequivalent.

AIM: To investigate the impact of oxycodone on gastrointestinal function in patients with moderate to severe acute respiratory distress syndrome undergoing mechanical ventilation. METHODS: Fifty patients with moderate to severe ARDS caused by severe pneumonia admitted to the Intensive Care Department of Fuling Hospital, Chongqing University from January 2022 to June 2023 were selected for the study. The patients were randomly divided into a remifentanil group (n=25) and a hydroxycodone group (n=25). They were treated with remifentanil and hydroxycodone for pain relief. We observed the incidence and severity grading of acute gastrointestinal injury (AGI), clinical outcomes (length of mechanical ventilation, ICU length of stay, and mortality at day 90), gastric emptying indicators (GRV, GER, MI), nutritional status (ALB, PAB), respiratory and circulatory indicators (Cst, OI, PaCO2, VIS). RESULTS: The occurrence of AGI showed no notable disparity between the remifentanil and oxycodone groups (96% vs. 72%, P=0.289), yet the remifentanil group exhibited a more severe AGI rating compared to the oxycodone group (P=0.009). The oxycodone groups GRV at 48 and 72 hours post-treatment was less than the remifentanil groups, with the MI surpassing the remifentanil groups (P<0.05), and the GRE exhibited comparable alterations (P>0.05). Post-treatment, the oxycodone groups ALB and PAB levels were elevated (P<0.05), whereas the remifentanil groups pre-treatment and post-treatment PAB remained unchanged (P>0.05), and the oxycodone groups PAB exceeded the remifentanil groups (P=0.035). However, the ALB, PAB, and ALB levels before and post-treatment were almost identical in both groups (P>0.05). Additionally, there was no significant variance in clinical and respiratory outcomes between the two patient groups (P>0.05). CONCLUSION: Hydroxycodone has a smaller inhibitory effect on gastric emptying in patients with moderate to severe acute respiratory distress syndrome undergoing mechanical ventilation, and has a milder degree of acute gastrointestinal injury. It can also improve the nutritional status of patients and has reliable safety. Hydroxycodone may be more beneficial in protecting the gastrointestinal function of patients with acute respiratory distress syndrome.

The salt-inducible kinase (SIK) family includes three isoforms, SIK1, SIK2 and SIK3, which are the main regulators of physiological and pathological processes and participate in the regulation of glucose and lipid metabolism, tumor, inflammation, depression, sleep-wake and circadian rhythm. SIK inhibitor has become a candidate drug for the treatment of a variety of diseases, and is especially expected to become a potential new drug for the treatment of nervous system diseases, such as sleep disorders, circadian rhythm disorders, and depression. This review summarizes the regulation and mechanism of three isoforms of SIK and their upstream and downstream signaling pathways in the above physiological and pathological processes. It also reviews the recentstudies of SIK inhibitors. 

Antibody drug conjugates (ADCs) are a new class of anti-tumor drugs in which linkers in the structure link cytotoxic drugs to monoclonal antibodies and release cytotoxic drugs to tumors. Disitamab vedotin (RC48) is a new antibody-drug conjugate independently developed in China. It targets the HER2 protein on the surface of tumors, it has both antibody targeting and small molecule drug killing, and can accurately recognize and kill tumor cells. Compared with traditional HER2-targeted drugs, disitamab vedotin has a wider therapeutic window and less toxicity to normal tissues. Currently, disitamab vedotin for injection has been approved by the National Medical Products Administration (NMPA) for use in patients with HER2 overexpression (HER2 immunohistochemical results of 2+ or 3+) who have locally advanced or metastatic gastric cancer (including gastroesophageal junction adenocarcinoma) and have received at least two types of systemic chemotherapy. Additionally, it is indicated for patients with locally advanced or metastatic urothelial carcinoma who have previously undergone platinum-containing chemotherapy and exhibit HER2 overexpression, specifically 2+ or 3+ immunohistochemical results. In this paper, we will review the structural characteristics, mechanism of action and clinical trials of disitamab vedotin and look forward to the clinical application prospects of this drug.