ObjectiveTo investigate the antitumor effect and mechanism of Guiqi Yiyuan ointment on Lewis lung cancer mice based on the extracellular regulatory protein kinase （ERK） signaling pathway. MethodsA Lewis lung cancer mouse model was established. Except for the blank group， the model mice were randomly divided into the model group， Guiqi Yiyuan ointment low， medium， and high dose groups， and the extracellular ERK1/2 inhibitor group， with 10 mice per group. The Guiqi Yiyuan ointment was administered by gavage at doses of 1.75， 3.5， 7.0 g·kg^-1·d^-1 for the low， medium， and high dose groups， respectively. The ERK1/2 inhibitor group was given the ERK1/2 inhibitor LY3214996 （100 mg·kg^-1·d^-1） by gavage. The treatment was administered for 14 consecutive days， after which samples were collected. Tumor histopathological changes were observed using hematoxylin-eosin （HE） staining. Transmission electron microscopy was used to observe ultrastructural changes in tumor cells. Immunofluorescence was performed to measure the phosphorylation of ERK1/2 （p-ERK1/2） and the expression of programmed cell death ligand-1 （PD-L1） in tumor tissues. Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were used to detect the expression of p-ERK1/2， PD-L1， the autophagy marker Beclin-1， the autophagic protein p62， and the microtubule-associated protein light chains LC3Ⅰ and LC3Ⅱ at both the protein and gene levels. ResultsCompared with the model group， the average tumor weight was significantly reduced in the low and medium dose groups of Guiqi Yiyuan ointment （P<0.05）， and markedly reduced in the high dose and inhibitor groups （P<0.01）. Tumor cells in all treatment groups became progressively irregular， with ruptured nuclei and expanded areas of cell disintegration and necrosis. The number of organellar ablations in tumor tissues increased， and the number of autophagic vesicles also increased in all groups. The mean fluorescence intensity of p-ERK1/2 and PD-L1 was reduced in the low and medium dose groups of Guiqi Yiyuan ointment （P<0.05）， and significantly reduced in the high dose and inhibitor groups （P<0.01）. The mRNA expression of ERK1/2， PD-L1， Beclin-1， and p62 was reduced in the medium dose group （P<0.05）， while LC3Ⅰ/Ⅱ mRNA expression was elevated （P<0.05）. In the high dose and inhibitor groups， mRNA expression of ERK1/2， PD-L1， Beclin-1， and p62 was significantly reduced （P<0.01）， while LC3Ⅰ/Ⅱ mRNA expression was significantly increased （P<0.01）. Protein expression of p-ERK1/2， PD-L1， Beclin-1， and p62 was reduced in the medium dose group （P<0.05）， and LC3Ⅰ/Ⅱ protein expression was elevated （P<0.05）. In the high dose and inhibitor groups， protein expression of p-ERK1/2， PD-L1， Beclin-1， and p62 was significantly reduced （P<0.01）， while LC3Ⅰ/Ⅱ protein expression was significantly elevated （P<0.01）. ConclusionGuiqi Yiyuan ointment may inhibit the activation of the ERK signaling pathway， downregulate the expression of p-ERK1/2， promote autophagic flux in tumor cells， and regulate the expression of PD-L1， thereby exerting an inhibitory effect on tumor growth in Lewis lung cancer mice.

Purpose To observe the clinical and CT features of severe immune checkpoint inhibitor-related pneumonitis(CIP)in lung cancer patients.Materials and Methods A total of 174 patients with lung cancer who received immune checkpoint inhibitor(PD-1/PD-L1 inhibitors)in Xi'an International Medical Center Hospital from September 1,2019 to March 31,2022 were retrospectively collected.Clinical and imaging features of patients with severe CIP were analyzed.Results There were 23 patients who met the diagnostic criteria of severe CIP.Among them,22 were male patients,15 were younger(＜65 years old),17 had a history of underlying lung disease,16 had a history of chemoradiotherapy and other treatments,and 21 had a history of combined radiotherapy and chemoradiotherapy.The median time from the initiation of immune checkpoint inhibitor to CIP was 128(74,348)days.19 patients were non-small cell carcinoma.CIP occurred in 16 patients with right lung cancer,15 had tumor central airway invasion,14 had radiographic features of diffuse alveolar injury/acute interstitial pneumonia pattern,and 20 died during follow-up.Conclusion Severe CIP is likely to occur in male lung cancer patients with a history of basic medical history and radiotherapy and chemotherapy.The clinical manifestations are varied,and the main imaging features are diffuse alveolar injury/acute interstitial pneumonia pattern,and the prognosis is poor.

Objective To explore the role of the SerpinB5 and β-catenin in occurrence and development of the primary hepatocellular carcinoma (HCC). Methods The expressions of SerpinB5 and β-catenin protein and mRNA in carcinoma tissues and paracancerous tissues of 60 patients with primary HCC were detected by immumohistochemistry and real-time quantitative reverse transcriptional polymerase chain reaction (RT-PCR) methods. Results The positive expression rate of SerpinB5 protein and SerpinB5 mRNA in carcinoma tissues were significantly lower than those in paracancerous tissues:25.0%(15/60) vs. 63.3%(38/60) and 1.12 ± 0.43 vs. 5.19 ± 0.39, and there were statistical differences (P<0.01). The positive expression rate of β-catenin protein and β-catenin mRNA in carcinoma tissues were significantly higher than those that in paracancerous tissues: 65.0%(39/60) vs. 31.7%(19/60) and 4.23 ± 0.25 vs. 1.19 ± 0.17, and there was statistical difference (P<0.01). Decreased SerpinB5 expression was associated with higher serumα-fetoprotein level, larger tumor size, poor differentiation, advanced TNM stage, capsule invasion and tumor thrombosis (P < 0.01 or 0.05). Increased β-catenin expression was associated with poor differentiation, advanced TNM stage, capsule invasion and tumor thrombosis (P < 0.01 or < 0.05). The correlation analysis result showed that SerpinB5 had negative correlation withβ-catenin (carcinoma issues:r=-0.346, P=0.001;paracancerous tissues:r=-0.258, P = 0.024). Conclusions The abnormal expression of SerpinB5 and β-catenin may contribute to the progression and biologically malignant behavior of primary HCC, and SerpinB5 and β-catenin exists synergistic effect in the occurrence and development of primary HCC.

OBJECTIVETo explore the safety and efficacy of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in preventing chemotherapy-induced neutropenia in patients with breast cancer and non-small cell lung cancer (NSCLC), and to provide the basis for clinical application.

METHODSAccording to the principle of open-label, randomized, parallel-group controlled clinical trial, all patients were randomized by 1∶1∶1 into three groups to receive PEG-rhG-CSF 100 μg/kg, PEG-rhG-CSF 6 mg, or rhG-CSF 5 μg/kg, respectively. The patients with breast cancer received two chemotherapy cycles, and the NSCLC patients received 1-2 cycles of chemotherapy according to their condition. All patients were treated with the combination chemotherapy of TAC (docetaxel+ epirubicin+ cyclophosphamide) or TA (docetaxel+ epirubicin), or the chemotherapy of docetaxel combined with carboplatin, with a 21 day cycle.

RESULTSThe duration of grade 3-4 neutropenia in the PEG-rhG-CSF 100 μg/kg and PEG-rhG-CSF 6 mg groups were similar with that in the rhG-CSF 5 μg/kg group (P>0.05 for all). The incidence rate of grade 3-4 neutropenia in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group, and G-CSF 5 μg/kg group were 69.7%, 68.4%, and 69.5%, respectively, with a non-significant difference among the three groups (P=0.963). The incidence rate of febrile neutropenia in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group and G-CSF 5 μg/kg group were 6.1%, 6.4%, and 5.5%, respectively, showing no significant difference among them (P=0.935). The incidence rate of adverse events in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group and G-CSF 5 μg / kg group were 6.7%, 4.1%, and 5.5%, respectively, showing a non-significant difference among them (P=0.581).

CONCLUSIONSIn patients with breast cancer and non-small cell lung cancer (NSCLC) undergoing TAC/TA chemotherapy, a single 100 μg/kg injection or a single fixed 6 mg dose of PEG-rhG-CSF at 48 hours after chemotherapy show definite therapeutic effect with a low incidence of adverse events and mild adverse reactions. Compared with the continuous daily injection of rhG-CSF 5 μg/kg/d, a single 100 μg/kg injection or a single fixed 6 mg dose of PEG-rhG-CSF has similar effect and is more advantageous in preventing chemotherapy-induced neutropenia.

Antineoplastic Agents ; adverse effects ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; Breast Neoplasms ; drug therapy ; Carboplatin ; administration & dosage ; adverse effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Cyclophosphamide ; administration & dosage ; adverse effects ; Epirubicin ; administration & dosage ; adverse effects ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Incidence ; Induction Chemotherapy ; Lung Neoplasms ; drug therapy ; Neutropenia ; chemically induced ; epidemiology ; prevention & control ; Polyethylene Glycols ; Recombinant Proteins ; administration & dosage ; Taxoids ; administration & dosage ; adverse effects

Objective To investigate the effects and mechanism of Sufentanil on Hep3B cell viability in liver cancer.Methods Sufentanil of different concentrations (0.01,0.1,1,5,10,1 5 μmol/L)was used to treat Hep3B cells.The changes of cell cycle,apoptosis and protein expression were detected to explore the potential mechanisms by MTT method,flow cytometry and Western blot,respectively.Results Reduced viability of Hep3B cells,arrested cell cycle in the G0/G1 phase,decreased expression of survivin protein,and increased expression of caspase-3 protein were observed with the increase of Sufentanil concentration. Conclusion Sufentanil inhibited the viability of Hep3B cells through affecting cell cycle,promoting cell apoptosis and changing protein expressions of survivin and caspase-3.

Female ; Gallbladder Neoplasms ; complications ; Humans ; Middle Aged ; Skin Neoplasms ; secondary

OBJECTIVETo detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC.

METHODSWe investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immunohistochemistry and real-time PCR.

RESULTSThe expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis.

CONCLUSIONAbnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.

Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; NF-E2-Related Factor 2 ; metabolism ; Neoplasm Staging ; Receptors, CXCR4 ; metabolism ; Signal Transduction

OBJECTIVETo observe autophagy induced by starvation in non-small cell lung cancer A459 and 95D cells.

METHODSA549 and 95D cells in logarithmic growth in 1640 medium were cultured in Earle's balanced salt solution (EBSS) for 0, 1, 2, 3, 4 or 5 h. Autophagosome formation in the cell culture was observed by MDC fluorescent staining, and the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 in the cells were detected using Western blotting.

RESULTSCompared with the control cells, the cells with prolonged starvation showed increased MDC-positive cells and autophagosome formation. The expression of Beclin-1 and the LC3-II/LC3-I ratio also increased as the starvation prolonged, reaching the peak levels at 3 h and 4 h, respectively.

CONCLUSIONAutophagy can be induced by starvation in A549 and 95D cells in correlation with the expression of autophagy-related proteins LC3 and Beclin-1. These cell models of nutritional deficiency-induced autophagy may allow for a better understanding of the role of autophagy in the development of non-small cell lung cancer.

Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Humans ; Membrane Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism

Objective To detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC. Methods We investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immumohistochemistry and real-time PCR. Results The expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis. Conclusion Abnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.

Objective To observe autophagy induced by starvation in non-small cell lung cancer A459 and 95D cells. Methods A549 and 95D cells in logarithmic growth in 1640 medium were cultured in Earle's balanced salt solution (EBSS) for 0, 1, 2, 3, 4 or 5 h. Autophagosome formation in the cell culture was observed by MDC fluorescent staining, and the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 in the cells were detected using Western blotting. Results Compared with the control cells, the cells with prolonged starvation showed increased MDC- positive cells and autophagosome formation. The expression of Beclin-1 and the LC3-II/LC3-I ratio also increased as the starvation prolonged, reaching the peak levels at 3 h and 4 h, respectively. Conclusion Autophagy can be induced by starvation in A549 and 95D cells in correlation with the expression of autophagy-related proteins LC3 and Beclin-1. These cell models of nutritional deficiency-induced autophagy may allow for a better understanding of the role of autophagy in the development of non-small cell lung cancer.