Aim To explore the effect of oxaliplatin combined with epidermal growth factor receptor tyrosine kinase inhibitor AG1478 on autophagy in non-small cell lung cancer H1975 cells. Methods H1975 cells were cultured in vitro using gradient concentrations of AG1478 (0, 5, 10, 15, 20, 25, 30, 35, 40 jjimol • IT

A high performance liquid chromatography (HPLC) method utilizing correction factors was established for the quantitative detection of related substances in flumazenil. Separation was achieved using an Agilent Pursuit XRs C18 column (250 mm × 4.6 mm, 5 μm) with an isocratic elution of dilute phosphoric acid, methanol, and tetrahydrofuran as the mobile phases. Correction factors calculated from a standard curve method were applied to determine the impurity content. The quantification of impurities in flumazenil was conducted using both external standard and correction factor methods, followed by validation and comparison of the two. For the identification of degradation products, a forced degradation approach was employed to prepare a flumazenil degradation solution, and the resulting impurities were confirmed by LC-MS analysis. The separation of flumazenil and its impurities was found to be efficient. The limits of quantification for impurities A, B, D, and E were established at 0.169 9, 0.314 7, 0.143 9, and 0.270 8 ng, respectively, with the limits of detection at 0.055 8, 0.096 9, 0.048 8, and 0.089 0 ng. These impurities demonstrated a strong linear relationship across the concentration ranges of 0.034 9-7.847 0, 0.038 7-8.710 7, 0.034 6-7.794 1, and 0.032 4-7.292 8 µg·mL^-1, respectively (n = 7). The method achieved average recoveries between 98.25% and 99.42%, with an RSD of less than 2.0% (n = 9), indicating high accuracy. The external standard and correction factor methods were used to determine the related substances in flumazenil, and the results of the two methods were consistent. The established HPLC method is characterized by its high accuracy, sensitivity, and repeatability, and is suitable for determining related substances in flumazenil.

Separation and determination of chiral and achiral impurities in glimepiride tablets by supercritical fluid chromatography. Chiral and achiral impurities were separated on a ACQUITY UPC² Trefoil^TM CEL1 column (150 mm × 3.0 mm, 2.5 μm) maintained at 30 ℃ with the mobile phase containing a mixture of CO₂ and methanol-isopropanol (1∶1) at 1 mL·min^-1, and the detection wavelength was set at 228 nm. The back pressure was set at 13.8 MPa. The injection volume was 5 μL. In the chromatogram of the system suitability solution, the peaks elute in the following order: impurity Ⅳ, impurity Ⅴ, glimepiride, impurity Ⅲ, impurity Ⅰ and impurity Ⅱ. The six substances were separated successfully in 6 min using the proposed method with a resolution factor of 2.9, 1.6, 3.0, 2.0, 6.4. The impurity Ⅰ-Ⅴ detection limit (S/N = 3) was 0.17, 0.10, 0.06, 0.15, 0.10 μg·mL^-1, respectively. Good linear relationship was established between the peak response and the concentration in the range of 0.48-51.30 μg·mL^-1 for all impurities. The spiked recovery of impurity Ⅰ-Ⅴ was found to be acceptable for 99.9%, 98.9%, 102.1%, 100.1%, 96.3% (n = 9), respectively. The related substance and assay results of 11 sample batches are consistent with the results obtained using the HPLC method in the Chinese Pharmacopoeia. Compared to the two HPLC methods in the Chinese Pharmacopoeia, the established supercritical fluid chromatography method can simultaneously separate glimepiride and its 5 impurities in a single run, and it has the following advantages: simplified sample preparation, greatly reducing the volumes of organic solvents, environmentally friendly, high accuracy and good reproducibility. It can be employed for the quality control of the chiral and achiral impurities in glimepiride tablets.

OBJECTIVE To analyze the pharmaceutical service process in a fracture patient complicated by fat embolism syndrome （FES） following postoperative fracture， aiming to provide a reference for clinical treatment and pharmaceutical service for similar patients. METHODS Clinical pharmacist participated in the entire treatment process of a patient with FES following postoperative fracture. Based on the patient’s clinical manifestations and test results， literature was reviewed to assist clinical physicians in formulating the therapeutic regimen of glucocorticoids. For the drug-related adverse reactions of renal function impairment and reduced platelet count that occurred during the treatment， suspicious drugs were analyzed and disposed of accordingly. RESULTS The clinical pharmacist recommended Hydrocortisone sodium succinate for injection （100 mg， q8 h， ivgtt， for about one week followed by a gradual dose reduction） for treating FES. The Vancomycin hydrochloride for injection used in this case was assessed as “very probably” associated with the adverse drug reactions of renal function impairment and thrombocytopenia. The clinical physician adopted the pharmacist’s medication recommendations， and the patient’s condition stabilized after treatment， with improvement in adverse reactions， and was discharged from the hospital. CONCLUSIONS The use of glucocorticoids in treating FES has a definite therapeutic efficacy. Clinical pharmacists should individualize the medication plan based on the patient’s pathological state and distinguish it from postoperative sepsis. Meanwhile， drug-induced adverse reactions in the kidney and blood system should be closely monitored.

Pyroptosis, a form of inflammatory cell death mediated by the Gasdermins family, promotes the release of inflammatory mediators and activates immune cell populations such as NK cells, T cells and macrophages in the tumor microenvironment(TME) to exert immune-regulating and anti-tumor effects. Glioblastoma(GBM) is the most serious and malignant glioma, and the median survival of patients diagnosed with GBM is less than 2 years, and the presence of the blood-brain barrier makes it difficult to deliver drugs to the brain, thus affecting the effect of drugs against GBM. Therefore, it is important to explore new measures and mechanisms to treat GBM, which has a complex TME with a large number of immune cell populations that are often immunosuppressed by GBM. Cellular pyroptosis as a mode of cell death capable of activating immunity, has the effect of activating the body’s immunity to help reverse TME immunosuppression. This review will focus on the relationship between cell pyroptosis and the immune system, how cell pyroptosis affects the immune cell population of TME in GBM, and the new progress in drug research on cell pyroptosis pathways in GBM treatment, providing new directions and strategies for future clinical treatment of GBM.

OBJECTIVE To investigate the effect of bs-5-YHEDA iron chelating peptide combined with semaglutide on the cognitive ability and pathological characteristics of D-Gal-induced Alzheimer's disease(AD) model mice. METHODS Forty mice were randomly divided into 5 groups, namely the healthy control group, PBS group, bs-5-YHEDA iron chelating peptide group, combined treatment group and positive control group, with 8 mice in each group, half of each sex. Except for the healthy control group, D-galactose was injected to induce the AD mice model for 6 weeks. For 3 consecutive weeks starting from the 4th week, the bs-5-YHEDA iron chelating peptide group was injected with bs-5-YHEDA(1 mg·mL–1) once every other day at 200 µL in the tail vein; the bs-5-YHEDA iron chelating peptide(1 mg·mL–1) and semaglutide(25 nmol·kg–1·d–1) were given alternately once a day in the combination treatment group; the positive control group was given memantine(3.3 mg·kg–1·d–1) by gavage every other day. The healthy control group and PBS group were injected with the equal dose of PBS. At the end of treatment, the learning memory ability of mice was detected by the Morris water maze method, whole brain and whole blood were dissected, and pathological changes in hippocampal region were observed by HE staining, and Aβ expression and Tau protein phosphorylation levels were detected by immunohistochemistry, enzyme-linked immunosorbent assay and immunoblotting. RESULTS In the Morris water maze spatial exploration experiment, the differences in the number of times the mice traversed the platform, the ratio of swimming distance to the target quadrant, and the time ratio were statistically significant in each group(P<0.05); compared with the PBS group, the ratio of swimming distance to the target quadrant increased in the combined treatment group, and the differences were statistically significant(P<0.05). The results of HE staining showed that compared with the healthy control mice, the hippocampal area in the PBS group showed reduced levels of pyramidal cells, disorganized arrangement, cell edema, and deep staining of nuclei consolidation. Cellular disorganization, deep staining of nuclei and apoptosis in the hippocampus were significantly improved in each treatment group after drug treatment. Immunohistochemistry and Western blotting results showed that the Aβ expression levels and Tau protein phosphorylation levels were significantly higher in the PBS-administered mice compared with the healthy control mice, and the Aβ expression levels and Tau protein phosphorylation levels were reduced in each group after drug treatment, with statistically significant differences(P<0.01 or P<0.001 ). CONCLUSION The combination of bs-5-YHEDA iron chelating peptide and semaglutide can effectively improve the learning and memory ability and pathological characteristics of AD mice, but from the results of immunohistochemistry and immunoblotting experiments, the improvement of pathological characteristics of AD mice in the combination treatment group is not obvious compared with the single bs-5-YHEDA iron chelating peptide group, suggesting that there may be a threshold effect of our designed dual-target combination treatment on the cognitive improvement of AD mice, and the optimization and validation of the effect of multi-target combination treatment need further study.

Objective To investigate the association between metabolic associated fatty liver disease(MAFLD)and carotid atherosclerotic plaque.Methods A total of 1 107 patients who were hospitalized in The Second Affiliated Hospital of Kunming Medical University from July,2014 to December,2022 were enrolled,and all patients underwent abdominal ultrasound and CT angiography of the head and neck arteries.Baseline data and clinical diagnosis were collected,and the patients were divided into MAFLD group with 499 patients and non-MAFLD group with 608 patients based on medical history,clinical tests,and imaging findings.According to the CT value,carotid plaques were classified into calcified plaques,non-calcified plaques,and mixed plaques.According to the NASCET criteria,carotid stenosis was categorized as normal vessel,slight stenosis,mild stenosis,moderate stenosis,and severe stenosis/occlusion.The independent-samples t test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups;the chi-square test was used for comparison of categorical data between two groups.Univariate and multivariate Logistic regression analyses were used to investigate the influencing factors for carotid atherosclerosis.Results Compared with the non-MAFLD group,the MAFLD group had a significantly higher proportion of patients with calcified plaques(74.3%vs 63.3%,P<0.05),non-calcified plaques(27.1%vs 17.1%,P<0.05),or mixed plaques(27.3%vs 20.7%,P<0.05),as well as a significantly higher proportion of patients with mild stenosis(50.9%vs 44.9%,P<0.05),moderate stenosis(14.6%vs 8.4%,P<0.05),or severe stenosis/occlusion(6.6%vs 3.5%,P<0.05).The univariate logistic regression analysis showed that MAFLD was a risk factor for calcified carotid plaques,non-calcified plaques,and mixed plaques,and it was also a risk factor for mild stenosis,moderate stenosis,and severe stenosis/occlusion of the carotid artery(all P<0.05).After adjustment for confounding factors,the multivariate Logistic regression analysis showed that MAFLD was an independent risk factor for calcified plaque,non-calcified plaque,mixed plaque,and moderate stenosis of the carotid arteries(all P<0.05).Conclusion MAFLD is an independent risk factor for moderate stenosis,calcified plaques,non-calcified plaques,and mixed plaques of the carotid arteries.

Objective To observe the value of vector flow mapping(VFM)technique for assessing changes of left ventricular diastolic function in ovarian cancer(OC)patients who underwent postoperative chemotherapy.Methods Totally 37 OC patients who received postoperative chemotherapy were prospectively enrolled in chemotherapy group,while 40 healthy adults were taken as controls(control group).Routine echocardiography and VFM were performed for chemotherapy group before chemotherapy,after 3 and 6 cycles of chemotherapy,also for controls at enrollment,and comparison was performed between groups before chemotherapy,as well as among different time points within chemotherapy group,and the correlations of VFM results with hemoglobin and routine echocardiographic results in chemotherapy group were analyzed.Results No significant difference of age,body mass,body surface area(BSA),nor hemoglobin level,routine echocardiographic and VFM results before chemotherapy was found between groups(all P＞0.05).With the process of chemotherapy,hemoglobin level gradually decreased,the isovolumic relaxation period(IR),atrial systole period(AS)intraventricular pressure difference(IVPD)and intraventricular pressure gradient(IVPG)of the left ventricle gradually increased(adjusted P＜0.05),whereas routine echocardiography only showed that the left atrial volume index(LAVI)and the ratio of early mitral inflow velocity and the mean mitral annular early diastolic velocity(E/e')increased after 6 cycles of chemotherapy compared with those pre-chemotherapy(adjusted P＜0.05).In chemotherapy group,VFM results in all diastolic subphases were strongly correlated with hemoglobin levels(|r|=0.718 to 0.836,all P＜0.05),weakly to moderately correlated with LAVI(|r|=0.375 to 0.525,all P＜0.05)and moderately correlated with E/e'(|r|=0.424 to 0.537,all P＜0.05).Conclusion The diastolic function of left ventricle was probably damaged in early stage after postoperative chemotherapy in OC patients.VFM might detect slight changes of early diastolic function of left ventricle more sensitively than routine echocardiography.

Objective To study the effects of radiation on the morphology,function,and NLRP3 expression of mouse salivary gland tissue,and provide new ideas to repair radiation-induced damage to salivary gland tissue.Methods We established a mouse model of radiation-induced submandibular gland injury and Recorded the weight of drinking water.The salivary flow rate was assessed,and HE staining was used to observe submandibular gland injury.Immunohistochemistry and Real-time PCR were used to assess expression of NLRP3 and Caspase-1 in the radiation-injured submandibular gland of mice at 1,3,7 and 14 days after radiation exposure.Results Over time,the amount of water consumed by radiation group mice was gradually increased,the salivary flow rate was decreased,and inflammatory cells in the submandibular gland continued to increase.Acinar cells gradually showed lesions,such as nuclear pyknosis and vacuolization.At 7 and 14 days after radiation exposure,expression levels of NLRP3 and Caspase-1 proteins and genes in the radiation group were significantly higher than those in the normal group(P＜0.05).Conclusions Radiation damages mouse submandibular gland tissue and activates the NLRP3 inflammasome to increase its expression level.

Aim To investigate the effect of tyrosine kinase inhibitor AG1478 combined with oxaliplatin (OXA) on apoptosis of colorectal cancer HCT116 cells. Methods MTT assay was used to measure the effect of AG1478 combined with OXA on proliferation of HCT116 cells. RT-qPCR was used to detect the mRNA expression levels of p53, caspase-3, Bcl-2 and Bax. Western blot was used to detect the proteins expression of p53, caspase-3, cleaved-caspase 3, Bcl-2, Bax, p62, LC3 and IL-6. Results Both OXA and AG1478 inhibited the proliferation of HCT116 (P < 0. 01). IC