Objective To investigate the role and primary mechanism of a novel fusion gene RELCH-RET in driving the malignant transformation of normal human bronchial epithelial(HBE)cells.Methods Based on retrospective clinical data from 456 non-small cell lung cancer(NSCLC)patients admitted in the Second Affiliated Hospital of Army Medical University from January 2019 to June 2022,a fusion gene,RELCH-RET,was identified as a research target.Three cell models were established:negative control(HBE VC,transfected with empty lentiviral vector),RET control(HBE RET,transfected with lentiviral overexpression vector of Flag-RET),and experimental group(HBE RELCH-RET,transfected with lentiviral overexpression vector of Flag-RELCH-RET).MTS assay and Transwell assay were used to detect cell proliferation and migratory and invasive abilities.In vivo tumorigenicity of the 3 cell models was assessed in 15 female non-obese diabetic/severe combined immunodeficiency(NOD/SCID)mice(SPF grade,4 weeks old,weighing 15.1±0.4 g)via subcutaneous xenograft experiments,with 5 animals in each group.Western blotting was employed to detect the autophosphorylation of RET(Y905)and the phosphorylation of downstream signaling proteins ERK1/2,EGFR(Y845)and STAT3(Y705).Dimerization and multimerization status of RELCH-RET were analyzed by chemical cross-linking(DTME treatment)in combination with Western blotting,with the reversibility being confirmed through de-cross-linking experiments.Results There were 3 cases carrying RELCH-RET fusion gene screened out from the 469 NSCLC patients.Compared with the HBE VC and HBE RET groups,the HBE RELCH-RET group exhibited significantly enhanced cell proliferation(P<0.01),and acquired migratory and invasive abilities(P<0.01),while the control groups did not demonstrate the abilities.In the mouse xenograft tumor model,HBE cells stably expressing RELCH-RET developed significant tumor nodules(P<0.001),whereas the control groups(empty vector and wild-type RET)failed to exhibit detectable tumor growth.Western blotting revealed that RELCH-RET could induce the autophosphorylation of the RET tyrosine residue(Y905)and significantly up-regulate the phosphorylation levels of ERK1/2,EGFR(Y845),and STAT3(Y705)proteins.Chemical cross-linking combined with Western blot analysis demonstrated that RELCH-RET formed a dimer(～170 kDa)in HBE cells,which is reversibly dissociated into monomers upon decross-linking treatment.Conclusion The novel fusion gene RELCH-RET,promotes ligand-independent dimerization/oligomerization,thereby mediating RET autophosphorylation,subsequently activates the downstream typical RET signaling pathway and ultimately drives the malignant transformation of normal HBE cells.

Objective To elucidate the effects of miR-616 on the malignant biological processes of osteosarcoma and to preliminarily explore its potential mechanisms.Methods In situ hybridization(ISH)was employed to analyze miR-616 expression in 11 paraffin-embedded osteosarcoma specimens collected in our department during January 2018 to December 2019.Quantitative real-time PCR(qRT-PCR)was used to compare the mRNA expression level of miR-616 in the osteoblast cell line hFOB1.19 and osteosarcoma cell lines 143B and HOS.Stable cell lines with miR-616 knockdown or overexpression were established via lentiviral transfection in 143B and HOS cells.Cell proliferation and apoptosis were detected by flow cytometry,while cell invasion and migration were assessed using Transwell and colony formation assays,respectively.To evaluate the effect of miR-616 on tumor growth in vivo,10 female nude mice(4 weeks old,weighing 18～20 g)were randomized into a control group and a miR-616 overexpression group.After the xenograft tumor model was constructed,the growth of subcutaneous tumors was monitored.Finally,next-generation sequencing and a dual-luciferase reporter assay were performed to identify the target genes of miR-616.Results ISH results showed that miR-616 expression was up-regulated in osteosarcoma tissues than adjacent tissues,and primarily localized in the cytoplasm.qRT-PCR confirmed that miR-616 level was significantly higher in 143B and HOS cells than hFOB1.19 cells(P<0.05).In vitro experiments revealed that miR-616 overexpression enhanced the proliferation,migration and invasion,while suppressing apoptosis in 143B and HOS cells(P<0.01).Conversely,miR-616 knockdown weakened these malignant phenotypes(P<0.05),with miR-616-3p showing a stronger effect on apoptosis than miR-616-5p.Animal experiments demonstrated that the tumor weight in the miR-616 overexpression group was significantly greater than that of the control group(98.00±17.22 vs 33.60±8.08 mg,P<0.01).Furthermore,KLF2 was identified and confirmed as a direct target of miR-616.Conclusion MiR-616 promotes malignant biological behaviors in osteosarcoma,and its expression level indicates that it may serve as a potential therapeutic target.

Objective To investigate the nutritional knowledge,attitudes,and behaviors of air crew in a sanatorium from 2020 to 2022,and analyzing their nutritional management methods.Methods A total of 118 air crew members in a sanatorium from 2020 to 2022 were included in this study.A questionnaire survey on nutritional knowledge,attitude and behavior was conducted to collect basic information.The scores of nutritional knowledge,attitude and behavior were compared among air crew of different types of job,ages,genders and diseases.Nutritional management methods were adopted according to the survey results,and the scores of nutritional knowledge,attitude and behavior were compared before and after nutritional management.Results Before nutritional management,the scores of nutrition knowledge,attitude and behavior of fighters were higher than those of transport planes and bombers(P<0.05);there were no significant differences in the nutritional knowledge,attitude or behavior scores between air crew members of different genders(P>0.05).After nutritional management,there were no significant difference in the nutritional knowledge,attitude,or behavior scores among air crew of different aircraft types,ages,genders,or disease conditions(P>0.05).The nutritional knowledge,attitude,and behavior scores were increased after nutritional management in all the air crew members(P<0.05).The nutritional knowledge,attitude,and behavior of air crew members were positively correlated with each other(P<0.05).Conclusion The air crew in a sanatorium from 2020 to 2022 have low scores of nutritional knowledge,attitude and behavior.Their knowledge,attitude,and behavior in nutrition management are mutually influenced.Nutrition management measures can improve their nutrition knowledge level,enhance their attitude towards nutrition management,and promote their healthy nutrition behavior.

The aim of this study is to construct an injectable gel with stable phototherapy and chemotherapy. Res-PTX@IR780 gel with phototherapy and chemotherapy property was prepared by introduction of photosensitizer IR780 and antioxidant resveratrol (Res) into the polyethylene glycol (PEG) solution of paclitaxel (PTX). The results showed that PTX, PTX@IR780 and Res-PTX@IR780 could form gels and the gels were injectable. ATR-FTIR results indicated not only components of the gels but also the formation of hydrogen bonding during the gelation. The results of UV showed instability of IR780 solution and stability improvement of Res-IR780 solution under infrared radiation (IR) irradiation. Photothermal tests showed that Res-PTX@IR780 displayed better photothermal conversion and photothermal stability under multiple irradiations than PTX@IR780. The results of
Animals ; Cell Line, Tumor ; Gels ; Hyperthermia, Induced ; Mice ; Mice, Inbred BALB C ; Nanoparticles ; Paclitaxel ; Phototherapy

OBJECTIVE： To study the effects of resveratrol （Res） on cognitive function and SIRT1/NF-κB signaling pathway in neonatal rats with hypoxic-ischemic brain injury. METHODS： SD neonatal rats were randomly divided into sham operation group （normal saline）， model group （normal saline）， Res low-dose and high-dose groups （30， 60 mg/kg）， with 12 rats in each group. Except that sham operation group received sham operation， hypoxic-ischemic brain injury model was established by Rice method in other groups. After modeling， the rats were given relevant medicine intraperitoneally each day， for consecutive 6 weeks. Water maze test was used to analyze spatial learning and memory function of rats in each group. The escape latency after 1， 3 and 6 weeks of administration and the times of crossing platform after 6 weeks of administration were recorded. TTC staining was used to detect cerebral infraction area of rats after 6 weeks of medication. Western blot was used to detect the expression of Bcl-2， Bax， Caspase-3， SIRT1， SIRT1/NF-κB pathway related protein SIRT1 and p-NF-κB in hippocampal CA1 region. RESULTS： Compared with sham operation group， escape latency of rats was prolonged significantly in model group after 1， 3， 6 weeks of medication （P＜0.05）， the times of crossing platform was decreased significantly after 6 weeks of medication （P＜0.05）； the area of cerebral infarction was increased significantly （P＜0.05）； the protein expression of Bax， Caspase-3 and p-NF-κB in hippocampus CA1 region were increased significantly， while the protein expression of Bcl-2 and SIRT1 were decreased significantly （P＜0.05）. Compared with model group， the escape latency of Res low-dose and high-dose groups were shortened significantly after 1， 3， 6 weeks of medication （P＜0.05）， while the times of crossing platform was increased significantly after 6 weeks of medication （P＜0.05）； the area of cerebral infarction was decreased significantly （P＜0.05）， and the protein expression of Bax， Caspase-3 and p-NF-κB protein in hippocampal CA1 area were decreased significantly， while the protein expression of Bcl-2 and SIRT1 were increased significantly （P＜0.05）. The improvement of above indexes in high-dose group were significantly better than low-dose group （P＜0.05）. CONCLUSIONS： Res can improve cognitive dysfunction in neonatal rats with hypoxic-ischemic brain injury， which is related with SIRT1/NF-κB signaling pathway.

The linear analysis for heart rate variability (HRV), including time domain method, frequency domain method and time-frequency analysis, has reached a lot of consensus. The non-linear analysis has also been widely applied in biomedical and clinical researches. However, for non-linear HRV analysis, especially for short-term non-linear HRV analysis, controversy still exists, and a unified standard and conclusion has not been formed. This paper reviews and discusses three short-term non-linear HRV analysis methods (fractal dimension, entropy and complexity) and their principles, progresses and problems in clinical application in detail, in order to provide a reference for accurate application in clinical medicine.
Electrocardiography ; Entropy ; Fractals ; Heart Rate ; Humans

Aim To study the effect of ginsenoside F1 on the enzyme activity and expression of gene of CYP3 A4 through activation of pregnane X receptor ( PXR ) . Methods With different concentrations of ginsenoside F1 treated on LS174T cells, the expression of CYP3A4 mRNA was determined by Q-PCR, and the enzyme activity was measured by P450-GloTM CYP3A4 assay according to the manufacturer′s instructions, fur-ther PXR-CYP3 A4 stable translation HepG2 cell lines were used to test ginsenoside F1 activates PXR by re-porter gene screening assay. Results The results re-vealed that the levels of CYP3 A4 gene and protein ex-pression were significantly increased by ginsenoside F1 in a concentration-dependent manner. At the same time, reporter gene screening showed that ginsenoside F1 could also enhance the transcriptional activity of PXR. Conclusion Ginsenoside F1 can significantly up-regulate the gene expression and enzyme activity of CYP3A4 via the PXR-CYP3A4 pathway.

Aim To study the induction effect of Ginkgolide B on CYP3A4,and further verify the role of pregnane X receptor in CYP3 A4 induction expres-sion. Methods With different concentrations of Ginkgolide B treatment on LS174T cells,the CYP3A4 mRNA expression was detected by Q-PCR assay,fur-ther PXR-CYP3 A4 stable translation HepG2 cell lines were used to test the effect of Ginkgolide B on activity of PXR by reporter gene screening assay.CYP3A4 protein expression was detected by Western blot.PXR was knocked down with transfected with siRNA, CYP3 A4 mRNA and protein were detected in the con-dition of PXR low expression.Results The results revealed that the level of CYP 3 A 4 gene and protein expression were significantly increased by Ginkgolide B,and there was no induction effect on PXR.Reporter gene screening showed that Ginkgolide B could en-hance the transcriptional activity of PXR in a concen-tration-dependent manner.Under conditions of low ex-pression of PXR ,Ginkgolide B could also increase ex-pression of CYP3A4,but the induction folds were low-er than those of normal PXR group.Conclusion Ginkgolide B can signicantly up-regulate CYP3 A4 ex-pression via the PXR-CYP3 A4 pathway,and it has no effects on PXR gene expression.

The paper is to report the study of the effect of Shenfu injection on the enzyme activity of liver CYP450 and its mRNA level of rat liver. Microsome of rat liver was prepared after intravenous administration of Shenfu injection for 7 days. The enzyme activity was quantified by Cocktail method. Meanwhile, the mRNA expression of CYP1A2, CYP2B1/2, CYP2C11 and CYP3A1 in the liver was detected by RT-PCR. Shenfu injection obviously induced the enzyme activities of CYP2B and CYP2C. Meantime Shenfu injection decreased the enzyme activities of CYP1A2 and CYP3A. The mRNA levels of CYP2B and CYP2C were also induced in rats treated with Shenfu injection. But it obviously inhibited the mRNA level of CYP1A2 and CYP3A. Since the enzyme activity and mRNA level were obviously changed after administration, the potential effect of drug-drug interaction should be concerned.

Objective To explore the pathological mechanism of brain function and structure in young patients with major depressive disorder by diffusion tensor imaging(DTI)and resting state functional magnetic resonance imaging(fMRI).Methods Sixteen participants diagnosed with major depression (MD) and 16 healthy age and gender-matched controls(HC) were recruited. Resting state fMRI and DTI brain scans were performed on all participants. A voxel-based method (VBM) was used to analyze the DTI datasets, and regional homogeneity (ReHo) approach was applied to preprocess the fMRI datasets. The value of fractional anisotropy (FA) and ReHo maps were obtained in the whole brain.Results FA values in the MD group were significantly lower than those of the healthy controls in the white matter of the left middle temporal gyrus, right middle frontal gyrus, left medial frontal gyrus, right precentral gyrus, left angular gyrus, left fusiform gyrus, left superior temporal gyrus, right middle temporal gyrus, right sub-gyral, left insula, and left pyramis (P＜0.01). ReHo in the MD group decreased in the left superior frontal gyrus, right superior frontal gyrus, left middle frontal gyrus, right middle frontal gyrus, left medial frontal gyrus, right medial frontal gyrus, left paracentral lobule, right paracentral lobule, right inferior parietal lobule, left postcentral gyrus, right postcentral gyrus, left middle occipital gyrus, left lingual gyrus, right putamen, right cingulate gyrus, right cuneus, left superior temporal gyrus, and right middle temporal gyrus (P＜0.01). Conclusion Abnormality of brain white microstructure and function exist widely in young patients with major depressive disorder. Abnormal connection of structures and function between the brain areas may be the key reason for the depression.