Objective To investigate the antitumor mechanism of astragaloside combine with cetuximab on the colon cancer cell line RKO of EGFR over expression through cell proliferation and autophagy. Methods The cell proliferation of colon cancer cell line RKO intervened by astragaloside with or without cetuximab was detected by 4- methyl- teerazolium (MTT). The expressive changes for protein of EGFR 、P62 and LC3 in RKO cells was detected by western bolt. Results MTT showed that 100 ug/mL astragaloside combined with 120 ug/mL cetuximab had significantly inhibiting effect on RKO cells of EGFR expression. Western blot showed that astragaloside can affect the expression of P62 and LC3 to suppress the occurrence ofautophagy. Conclusion In the vitro studies showed that astragalosidecan enhance the antitumor cell proliferation of cetuximab through inhibit autophagy in the treatment of colon cancer.

Objective　To observe the effect of nordihydroguaiaretic acid treatment combined with Methotrexate or Vincristine on a human malignant glioma cell line. Methods　The effect of drugs on the cells was measured by MTT, and the protein expression of cyclin D1 gene in the cells was measured by immunohistochemistry assay. Results　①The anti-tumor effects of drugs on the cells were increased with the increasing of concentrations, and had a combined function between NDGA and MTX or VCR. ②There was no difference between the effect of treating NDGA with MTX or VCR given at the same time and MTX or VCR given after 24 h of NDGA treatment, but the effects were apparently different when NDGA was administrated after MTX or VCR treatment for 24 h. ③The protein expression of cyclin D1 gene in the cells was apparently reduced after NDGA treatment. Conclusion　There is a cooperative function between NDGA and MTX or VCR on the cells, and the function is probably related to the NDGA-induced down-regulation of protein expression of cyclin D1 gene in the cells.

Objective To identify the morphological parameters that are related to intracranial aneurysms (IAs) rupture using a case-control model. Methods A total of 107 patients with multiple IAs and aneurysmal subarachnoid hemorrhage between August 2011 and February 2017 were enrolled in this study. Characteristics of IAs location, shape, neck width, perpendicular height, depth, maximum size, flow angle, parent vessel diameter (PVD), aspect ratio (AR) and size ratio (SR) were evaluated using CT angiography. Multiple logistic regression analysis was used to identify the independent risk factors associated with IAs rupture. Receiver operating characteristic curve analysis was performed on the final model, and the optimal thresholds were obtained. Results IAs located in the internal carotid artery (ICA) was associated with a negative risk of rupture, whereas AR, SR1 (height/PVD) and SR2 (depth/PVD) were associated with increased risk of rupture. When SR was calculated differently, the odds ratio values of these factors were also different. The receiver operating characteristic curve showed that AR, SR1 and SR2 had cut-off values of 1.01, 1.48 and 1.40, respectively. SR3 (maximum size/PVD) was not associated with IAs rupture. Conclusions IAs located in the ICA are associated with a negative risk of rupture, while high AR (>1.01), SR1 (>1.48) or SR2 (>1.40) are risk factors for multiple IAs rupture.

BACKGROUNDTo evaluate the relationship between estrogen receptor (ER) expression and level of serum sexual hormones in male patients with lung cancer.

METHODSThe levels of serum estradiol (E 2), testosterone (T), follicle stimulating hormone (FSH) and lutenising hormone (LH) were measured in 25 patients with lung cancer and 30 healthy men by enzyme immunoassay magnetic solid phase (IEMA), the ER expression was detected in 25 cancer tissues, 25 paracancerous tissues, and 11 benign pulmonary tissues by immunocytochemistry (ICC).

RESULTSThe level of plasma E₂ in male patients with lung cancer was significantly higher than that in normal controls [(22.4±15.7) ng/L [WTBX]vs ( 12.6±4.8) ng/L, P=0.001 9] while the level of T was significantly lower while the level of T was significantly lower [(2.9±1.3) μg/L [WTBX]vs (4.1±1.5) μg/L, P= 0.003 0]. The ratio of E₂/T of male patients with lung cancer was also remarkably higher than that of control . The ratio of E₂/T of male patients with lung cancer was also remarkably higher than that of control [(9.7±10.0) ×10⁻³[WTBX]vs ( 3.4±1.6)×10⁻³, P=0.004 6]. The expression rate of ER in lung cancer tissue samples was 60.0% (15/25), but no ER expression was found in the paracancerous tissues and benign pulmonary tissues. The level of E₂ had positive correlation with the expression of ER in male patients with lung cancer (. The expression rate of ER in lung cancer tissue samples was 60.0% (15/25), but no ER expression was found in the paracancerous tissues and benign pulmonary tissues. The level of E₂ had positive correlation with the expression of ER in male patients with lung cancer (r=0.916 7, P < 0.001).

CONCLUSIONSThere are disorders and imbalances of sexual hormone metabolism in male patients with lung cancer, and these imbalances relate to the expression of ER. The elevation of E₂ level in peripheral blood might be related to the overexpression of ER.

BACKGROUNDTo improve the efficacy and selectivity of gene therapy for lung cancer through inducing oncostatin M (OSM) gene expression by radiation via the early growth response gene-1 (Egr-1) promoter.

METHODSThe radio-inducible OSM gene was constructed by insertion of Egr-1 promoter into upstream of the OSM gene. The expression of OSM in lung adenocarcinoma cell line A549 which was transfected with pEO and exposed to different doses of γ-ray irradiation was analyzed, and the relative survival fraction of cells and cell survival curve were observed. To examine the efficacy of this pEO gene therapy in vivo, the tumor supression effects were investigated in 40 nude mice bearing lung tumors.

RESULTSExpression of OSM gene in A549 cells transfected with pEO plasmids was markedly upregulated in a radiation dose-dependent manner. A gene therapy experiment in vitro showed that pEO transfected A549 cells became highly sensitive to ionizing radiation. pEO transfected tumors regressed significantly after a combination therapy with irradiation in all mice (n=10), and three tumors disappeared in 3 weeks without any side effect.

CONCLUSIONSThe results indicate that tumor targeted expression of OSM gene under the control of a radio-inducible promoter represents a novel strategy for safe and effective gene therapy for lung cancer and might be widely applied in the future.

Objective:To investigate whether the lung specific X protein(LUNX)gene can serve as a prognostic biomarker for non-small cell lung cancer related to immune cell infiltration.Methods:A total of 280 non-small cell lung cancer patients admitted to Hengshui People's Hospital from January 2020 to January 2023 were selected to detect the expression of LUNX gene in cancer tissue and adjacent tissues,and to analyze the relationship between LUNX gene and immune cell infiltration and prognosis survival status in the tumor microenvironment.Results:Compared with adjacent tissues,the expression level and positive rate of LUNX gene in non-small cell lung cancer tissue were increased,which were related to differentiation degree,lymph node metastasis and tumor staging(P<0.05).GEPIA database analysis showed that the LUNX gene was only slightly expressed or not expressed in other tissues,while its expression was elevated in LUAD and LUSC(P<0.05).The copy number of LUNX gene and LUNX gene were related to the level of immune cell infiltration(P<0.05).Survival analysis showed that high expression of the LUNX gene was associated with patient survival prognosis(P<0.05).Conclusion:The LUNX gene is specifically expressed in non-small cell lung cancer tissue,affecting the level of immune cell infiltration in non-small cell lung cancer,leading to an imbalance in the immune microenvironment,and is an important mechanism for causing patients prognostic death,which can be used as a prognostic biomarker for evaluating immune cell infiltration.

Objective To investigate the correlation between serum Irisin,long pentraxin 3(PTX3),human metas-tasis-associated lung adenocarcinoma transcript 1(MALAT1)levels and the severity of diabetic retinopathy(DR)and the value of combined diagnosis.Methods Eighty-five patients with type 2 diabetes mellitus(T2DM)combined with DR at Cangzhou Central Hospital from April 2022 to April 2023 were selected as the DR group,85 patients with T2DM alone were selected as the non-DR group,and 85 healthy volunteers were selected as the control group during the same period.Pa-tients in the DR group were further divided into the proliferative DR(PDR)group(38 patients)and the non-PDR group(47 patients)based on whether DR was in the proliferative phase.Clinical data of patients in the DR group were collected,including gender,diastolic pressure,age,systolic pressure,disease course,fasting plasma glucose(FPG),body mass in-dex,hemoglobin A1c(HbA1c),smoking history,triglyceride(TG),drinking history,peak systolic velocity(PSV),peak end-diastolic velocity(PEDV),resistance index(RI),fasting insulin(FINS),family history of diabetes,total cholesterol(TC),and homa-insulin resistance(HOMA-IR).Enzyme-linked immunosorbent assay was used to detect serum levels of Irisin and PTX3 in each group of patients,and real-time quantitative polymerase chain reaction was used to detect the ser-um level of MALAT1.The correlations between serum levels of Irisin,PTX3 and MALAT1 and the severity of DR were ana-lyzed using the Pearson correlation coefficient.The influencing factors of the DR severity were identified using the Logistic regression.The value of serum Irisin,PTX3,and MALAT1 levels in diagnosing DR alone was analyzed using the receiver operating characteristic(ROC)curve.The value of regimens containing and not containing serum Irisin,PTX3,and MAL-AT1 levels in diagnosing DR was analyzed using the ROC curve,net reclassification index(NRI),and integrated discrimina-tion improvement(IDI)index.Results The serum levels of Irisin,PTX3,and MALAT1 were compared among the three groups of patients,and the differences were statistically significant(all P＜0.001).The disease course of patients in the PDR group was longer than that in the non-PDR group,the PSV,PEDV and serum Irisin level were lower than those in the non-PDR group,while the RI,FPG,HbA1c,TG,FINS,HOMA-IR,and serum PTX3 and MALAT1 levels were higher than those in the non-PDR group(all P＜0.05).The serum Irisin level in DR patients was negatively correlated with the severity of DR(r=-0.512,P＜0.001),while the PTX3 and MALAT1 levels were positively correlated with the severity of DR(r=0.497,0.573,both P＜0.05).The Logistic regression analysis showed that the disease course,FPG,HbA1c,TG,FINS,HOMA-IR,PSV,PEDV,RI,and serum levels of Irisin,PTX3 and MALAT1 were influencing factors for the DR progression(allP＜0.05).The area under the curve(AUC)of serum Irisin,PTX3,and MALAT1 levels in diagnosing DR was 0.743,0.811,and 0.773,respectively.Compared with conventional diagnostic protocols,the AUC of the new diagnostic protocol containing serum levels of Irisin,PTX3,and MALAT1 significantly increased(Z=2.708,P=0.007),and the NRI and IDI were 0.039(95％CI:0.022-0.069)and 0.026(95％CI:0.014-0.047),respectively(all P＜0.05).Conclusion The serum Irisin level in DR patients decreases,while the serum PTX3 and MALAT1 levels increase,which are closely related to the severity of DR.Diagnostic plans containing serum Irisin,PTX3,and MALAT1 indicators have high diagnostic value.

ObjectiveTo investigate the mechanism of Huazhuo Jiedu Huoxue Tongluo prescription in alleviating cerebral ischemia-reperfusion injury via regulating nerve cell autophagy based on c-Jun N-terminal kinase（JNK）signaling pathway . MethodSixty SD rats were randomly divided into 6 groups： sham group， middle cerebral artery occlusion/reperfusion （MCAO/R） group （model group）， Huazhuo Jiedu Huoxue Tongluo prescription group ［traditional Chinese medicine （TCM） group（25.0 g·kg^-1）］， JNK inhibitor SP600125 （SP） group（5 mg·kg^-1）， TCM+SP group and JNK agonist Anisomycin （Ani） group（15 mg·kg^-1）. After 24 h of modeling， TCM group and TCM+SP group were given TCM decoction （ig） for 3 consecutive days， and the other groups were given equal volume of normal saline （ig）. Neurological deficit was evaluated by neurological function score and cerebral infarct volume was determined by 2，3，5-triphenyltetrazole chloride （TTC） staining. Hematoxylin-eosin （HE） staining and Nissl staining were used to observe the structural changes of brain tissue and the damage of neurons， respectively. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay （TUNEL） was performed to detect cell apoptosis. The ultrastructure of autophagosomes was observed by transmission electron microscope. Western blot was employed to detect the protein expressions of microtubule-associated protein 1 light chain 3A/B （LC3A/B）， autophagy related 5 （Atg5）， the ortholog of yeast Atg6 （Beclin1）， p62， B-cell lymphoma 2 （Bcl-2）， JNK， phosphorylated （p）-JNK and c-Jun in brain tissue. The mRNA expressions of LC3A/B， Beclin1， p62， Atg5， Bcl-2， JNK and c-Jun were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the sham group， the model group had elevated neurological deficit score （P<0.05）， enlarged cerebral infarct volume （P<0.05）and typical infarction manifestations formed in hippocampal region and its surrounding brain tissue. In addition， there were a large number of neuronal cell degeneration， necrosis， liquefaction， nucleus pyknosis and deep staining， and inflammatory cell infiltration in the cortex in the model group， and severe swelling of mitochondria， lysosomes， autophagosomes and autophagolysosomes were clearly seen under electron microscope. TUNEL positive cells were increased （P<0.05）， and cell apoptosis was severe. The nuclear membrane and nucleolus of neurons in brain tissue were blurred with discontinuous processes， and Nissl bodies in cytoplasm were stained light with reduced number. Western blot revealed that the model group had up-regulated protein expressions of LC3A/B， Beclin1， Atg5， JNK， p-JNK and c-Jun in brain tissue （P<0.05）， while down-regulated protein expressions of p62 and Bcl-2 （P<0.05）as compared with the sham group. Real-time PCR indicated that the mRNA expressions of LC3A/B， Beclin1， Atg5， JNK and c-Jun in the model group were higher （P<0.05） while the mRNA expressions of p62 and Bcl-2 were lower （P<0.05） than those in the sham group. Compared with the conditions in model group， the neurological deficit scores of TCM， SP and TCM+SP groups were lowered （P<0.05）， and the cerebral infarct volume was reduced （P<0.05）， with improved pathological status of brain tissue， especially in the TCM group. Furthermore， there were abundant and basically normal mitochondrial cristae， slightly dilated endoplasmic reticulum， slightly swollen golgi apparatus， slightly fused nuclear membrane， and few visible lysosomes， autophagosomes and autophagolysosomes. TUNEL positive cells were decreased （P<0.05）， displaying reduced apoptosis， especially in the TCM group. The nucleolus and nuclear membrane of neurons in brain tissue were discernible， and Nissl bodies in cytoplasm was reduced to a certain degree as compared with those in the model group. Western blot showed a decrease in the protein expressions of LC3A/B， Beclin1， Atg5， JNK， p-JNK and c-Jun in the TCM group ，the SP group，and the TCM+SP group（P<0.05），while an increase in the protein expressions of p62 in the TCM group and SP group（P<0.05），and an increase in the protein expressions of Bcl-2 in the TCM group and TCM+SP group. By Real-time PCR， the mRNA expressions of LC3A， LC3B， Beclin1， Atg5， JNK and c-Jun had a down-regulation（P<0.05） while the mRNA expression of p62 a up-regulation in the TCM group ，the SP group，and the TCM+SP group（P<0.05），and the mRNA expression of Bcl-2 a up-regulation in the TCM group and the TCM+SP group（P<0.05）.Scores of the Ani group were raised （P<0.05）， and infarct volume was increased significantly， with aggravated neuronal cell necrosis and obvious inflammatory infiltration. Moreover， there were neuronal nuclear membrane fusion with abnormal protrusion， increased heterochromatin aggregation in edge， severe mitochondrial swelling， endoplasmic reticulum expansion， increased lysosomes， increased intracytoplasmic vesicles， and visible autophagosomes and autophagolysosomes. TUNEL positive cells were increased （P<0.05）， representing severe apoptosis. The number of Nissl bodies dropped with light staining， and the nucleolus and nuclear membrane were blurred. Real-time PCR found that the mNRA expressions of Atg5， c-Jun， JNK were up-regulated （P<0.05），while Beclin1， p62， Bcl-2 were were down-regulated in the Ani group （P<0.05）. Compared with the TCM group and SP group，the protein expressions of LC3A/B， Beclin1， Atg5， JNK， p-JNK， c-Jun were decreased，and p62， Bcl-2 were increased in the Ani group（P<0.05）. Compared with the TCM group，the mRNA expressions of JNK mRNA had a down-regulation in the SP group and TCM+SP group，while LC3A， LC3B， Atg5， c-Jun， JNK had an up-regulation（P<0.05） and Bcl-2 had a down-regulation in the Ani group（P<0.05）. Compared with the SP group，the mRNA expressions of Atg5， c-Jun， JNK had an up-regulation（P<0.05）， and Beclin1， p62， Bcl-2 had a down-regulation in the Ani group（P<0.05）. ConclusionHuazhuo Jiedu Huoxue Tongluo prescription significantly up-regulates the protein and mRNA expressions of LC3A/B， Beclin1， Atg5， JNK， p-JNK and c-Jun， and down-regulates the protein and mRNA expressions of p62 and Bcl-2， suggesting that the prescription can inhibit autophagy through JNK signaling pathway to reduce ischemia/reperfusion injury in rats.