Objective To investigate the proliferation inhibition and the apoptosis induction effect of brucine on human chronic myeloid leukemia cell line HL-60 cells.Methods HL-60 cells were exposed to various dosages of brucine 24,48,72 h respectively,MTT method was used to assay the growth inhibition effect of brucine on HL-60 cells and the IC50 of brucine was evaluated at the same time.The morphology was observed by AO/EB stains.The cell apoptosis and cell cycle was tested by flow cytometry with Annexin V-FITC/PI double staining and PI labeling respectively.Results The results indicated that the brucine displayed significant anti-proliferative effect on HL-60 cells in a dose-and time-dependent manner,with IC50 value of 211.8 μg/ml(24 h),107 μg/ml(48 h),83 μg/ml(72 h)respectively.The most significant inhibition was observed at 320 μg/ml for 48 h.In this condition,apoptosis morphology was induced by brucine with nuclear chromatin condensation,most of the nuclears were orange stained and condensation-like or bead-like,which was consistent with the Annexin V-FITC/PI results.The cell apoptosis rates were(2.1±1.1)％,(21.3±1.2)％,(38.6±1.3)％,(58.5±4.1)％,(75.3±0.87)％and(66.2±0.75)％in different dose of brucine,respectively.At the same time,the cell cycle analysis results showed that the cell ratio in G0/G1 phase was decreased while in G2/M and sub-G0 phases was increased,comparing with blank control group.Conclusion Brucine can inhibit cell growth dramatically,which may be related to the cell apoptosis and the cell cycle arrest.

Objective To prepare and identify rabbit polyclonal antibody against embryonic liver fordrin 3(ELF3),and investigate the distribution of ELF3 in mice tissue.Methods ELF3 specific N-terminal peptide was synthesized,and conjugated to Keyhole limpet hemocyanin(KLH)as immunogen.The ELF3-KLH complex was injected into rabbits subcutaneously,and then ELF3 antibody was purified using affinity chromatography.The titer of the antibody was evaluated by ELISA.The specificity of antibody against ELF3 and immunolocalization of ELF3 were evaluated by using Western blot and immunohistochemistry.Results Rabbit polyclonal antibody against ELF3 was prepared by the immunization of ELF3-KLH complex.ELISA and Western blot results showed the antibody against ELF3 had high titer and specificity.Western blot and immunohistochemical studies demonstrated ELF3 was expressed in the mouse heart,liver,brain and kidney tissue,particular on the cell membrane.Conclusion The preparation of polyclonal antibody against ELF3 was successful due to its high titer and specificity;ELF3 was expressed in the mice heart,liver,and kidney,particular on the cell membrane.It will provide an excellent tool for further study on the ELF3 function.

For the treatment of liver tumors,interventional therapy is becoming more and more important.The development of interventional therapy relies on the development and application of new materials,new equipment and new drugs,and the preclinical animal testing of these applications is an important part of assessing the safety and effectiveness of interventional procedures.The animal HCC model plays an important leading role in exploring the new approach of interventional therapy.However,the animal HCC models which are suitable for experimental researches are limited,at present,the animal HCC models are mainly confined to rats and rabbits.Due to the limitation of animal body shape,routine interventional instruments and equipment are not suitable to perform interventional procedures for some small animals.Nowadays,there is still a shortage of devices for interventional hepatic artery surgery in mice,and interventional instruments for rats are also limited.This paper aims to make a summary about the types of HCC model in rats and rabbits which are often used in the experimental researches,the modeling methods,the interventional procedures,the anesthesia and imaging examinations,etc.

Objective To investigate the effects of crude extract of Capparis spinosa L. fruit alka-loids (CSFA) on the maturation of murine bone marrow-derived dendritic cells (DCs). Methods CSFA was prepared and the contents were determined by high performance liquid chromatography. DCs were trea-ted with different doses (1, 2, 3 mg/ml) of CSFA. The viability of DCs, the expression of surface mole-cules and the ability of phagocytosis were detected by flow cytometry. The secretion of cytokines was meas-ured by ELISA. Western blot assay was performed to analyze the activation of key molecules in mitogen-acti-vated protein kinases ( MAPK) and nuclear factor-kappa B ( NF-κB) signaling pathways. Results The re-sults showed that CSFA alone had no significant influence on the expression of surface molecules and cyto-kines in DCs. However, it significantly decreased the expression of CD40, CD80, CD86 and MHC Ⅱ as well as the secretion of IL-12p40 and TNF-αthat were induced by lipopolysaccharides (LPS), but increased IL-10 secretion and the ability of phagocytosis after treating DCs with both CSFA and LPS. Further, the phosphorylation of p38, extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) and the nuclear translocation of NF-κBp65 induced by LPS were inhibited by CSFA. Conclusion CSFA could inhibit the maturation of DCs and the secretion of pro-inflammatory cytokines induced by LPS while in-creasing the secretion of the anti-inflammatory cytokine IL-10 and the ability of phagocytosis, which might in-volve MAPK and NF-κB signaling pathways. This study suggests that CSFA could be used as a potential im-munosuppressant.

Objective:To investigate expression level of transcriptional co-activator with PDZ-binding motif(TAZ) in neonatal rats with chronic lung diseases induced by hyperoxia and explore its potential role in the disease.Methods:The model of high-oxygen-induced lung injury in neonatal rats was established by continuous inhalation of high-concentration oxygen.The rats in experimental group inhaled 85% oxygen, while the rats in control group inhaled air.The lung tissues were collected at the 1st, 3rd, 7th, 14th and 21st day, and the lung tissue sections were stained with hematoxylin and eosin staining to observe the pathological changes of the lung.In addition, the dynamic expressions of TAZ, surfactant protein C(SPC) and aquaporin-5(AQP5) in lung tissue were detected by real-time PCR, western blot and immunohistochemistry staining.Results:In the experimental group, with the prolongation of oxygen inhalation time, we found that the alveolar cavity increased, the number decreased, the alveolar septum thickened, and the alveolar structure was simplified.Compared with the control group, there was no difference in TAZ, SPC and AQP5 expression at 1st and 3rd days in the lung tissue in the experimental group( P>0.05). However, at 7, 14 and 21 days, the expression of TAZ in mRNA and protein level in lung tissue in experimental group decreased significantly, and the expression of SPC in mRNA and protein level increased significantly, while the expression of AQP5 in mRNA and protein level decreased, the differences were all statistically significant( P<0.05). Conclusion:Hyperoxia can cause alveolar structure disorder and pulmonary arrested development in neonatal rat.The expression levels of SPC and AQP5 show that the injury of type Ⅰ alveolar epithelial cells (AEC Ⅰ) is severe.Although the number of type Ⅱ alveolar epithelial cells (AEC Ⅱ) increased, but its differentiation capacity decreased obviously.The decrease of TAZ expression may cause AEC Ⅱ lose the function of differentiation into AEC Ⅰ.

Objective:To investigate the expression of long non-coding RNA metastasis associated lung adenocarcinoma transcript 1(lncRNA MALAT1) in bronchopulmonary dysplasia (BPD) of neonatal rats induced by hyperoxia and its effect on alveolar type 2 epithelial cells (AEC Ⅱ).Methods:The lung injury model of neonatal SD rats induced by hyperoxia(model group, n=50, inhaled oxygen concentration of 80%-85%) and the control group(inhaled air, n=50) were prepared.Lung tissue samples were taken and retained on days 1, 3, 7, 14 and 21, and the physiological and pathological changes of lung tissue were detected by paraffin-embedded sections and hematoxylin-eosin staining; The dynamic expression of lncRNA MALAT1 in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction; The dynamic expression of surfactant protein C(SPC) in lung tissue and AECⅡ was detected by Western blot.AECⅡ was extracted from lung tissue of normal newborn rats, and lncRNA MALAT1 was knocked down by siRNA.The cells were collected and Western blot as well as immunofluorescence were used to detect the changes of SPC. Results:The lung tissue of model group gradually became thickened with alveolar compartments, and the alveolar cavity was enlarged with the disappearance of alveolar spine and other pathological structural changes.Compared with the control group, there was no difference in the expression of lncRNA MALAT1 and SPC in the lung tissue from model group on days 1, 3( P>0.05), but the expression of lncRNA MALAT1 and SPC significantly increased on days 7, 14 and 21( P<0.05). When lncRNA MALAT1 was inhibited, SPC expression showed a decrease trend. Conclusion:Hyperoxia can lead to the stagnation of lung development in neonatal rats, and the structure and function of alveolar disorders are impaired.The expression of lncRNA MALAT1 is involved in the process of hyperoxia-induced BPD in neonatal rats.The increase of lncRNA MALAT1 may promote the proliferation of AECⅡ.

Objective To investigate the effects of Artemisia absinthium L. ( A. absinthium L. ) extracts on the maturation and function of dendritic cells (DCs). Methods A. absinthium water (AAW) and ethanol ( AAE) extracts were prepared. DCs were separated into several groups and treated with differ-ent concentrations of AAW (containing 5, 50 and 150μg/ml of polysaccharide) and AAE (containing 0. 6, 3 and 6 μg/ml of flavonoid) , respectively. Cell viability, antigen phagocytosis and maturation of DCs were detected by flow cytometry. Levels of cytokines were analyzed by ELISA. Western blot assay was performed to analyze the activation of key molecules in NF-κB, mitogen-activated protein kinase ( MAPK) and janus kinase/signal transducer and activator of transcription ( JAK/STAT) signaling pathways. Results AAW promoted the maturation of DCs, significantly decreased antigen phagocytosis and increased cytokine produc-tion (IL-12p40 and TNF-α). AAE significantly enhanced the expression of co-stimulatory molecules on the surface of DCs and decreased antigen phagocytosis, but had no significant effect on cytokine production. Mo-reover, AAE significantly inhibited the LPS-induced expression of TNF-α, IL-12p40 and IL-6. Further anal-ysis revealed that AAW and AAE could activate the phosphorylation of p38, extra-cellular signal regulated kinase (ERK), IKKα/β, NF-κBp65 and JAK2. Besides, AAE could activate the phosphorylation of c-Jun N-terminal kinase ( JNK ) and inhibit the LPS-induced phosphorylation of inhibitor of NF-κB ( IκB-α) , IKKα/β, NF-κBp65, p38, ERK and JAK2. Conclusion AAW could enhance immunity and AAE could inhibit inflammation.

Objective To investigate the effects of ethanol extracts of Lepidium meyenii Walp (LMEE) from two different areas in Xinjiang on the maturation of mouse macrophages (RAW264. 7 cells) and dendritic cells (DCs). Methods Ethanol extracts of LMEE from Tashikuergan County (Ta xian) and A La gou of Xinjiang were prepared and named as LMEE-T and LMEE-A, respectively. RAW264. 7 cells and bone marrow-derived DCs from C57BL/6 mice were treated with different concentrations of LMEE-T/A. The viability of RAW264. 7 cells was analyzed by MTT assay. Expression of costimulatory molecules and MHCⅠ on the surface of RAW264. 7 cells and DCs was detected by flow cytometry. Secretion of cytokines and the release of nitrogen oxide (NO) were measured by ELISA and Griess method, respectively. Results LMEE-T/A had no significant influence on the viability of RAW264. 7 cells when the concentration was lower than 1 mg/ml. Treating RAW264. 7 cells with LMEE-T/A promoted surface molecule expression, cytokine secretion and NO release through TLR4 signaling pathway in a dose-dependent manner. Moreover, LMEE-T was more potent than LMEE-A. LMEE-T/A at the concentration of 0. 4 mg/ml promoted the expression of DC surface molecules and the secretion of cytokines. Infrared and ultraviolet spectra showed that LMEE-A and LMEE-T contained polysaccharides, macaenes, macamides and flavanols. Compared with LMEE-A, LMEE-T contained more benzene ring compounds but less polysaccharides. Conclusion Both LMEE-T and LMEE-A could activate RAW264. 7 cells and promote the maturation of DCs. The differences between their effects might be related to the differences in their contents.

Objective To establish and validate a voxel-based method for the quantitative detection of air trapping (AT),and to explore its diagnostic value by preliminarily apply this method in chronic obstructive pulmonary disease (COPD) patients.Methods From March 2015 to February 2016,fifty healthy young volunteers and eighteen COPD patients who underwent both end-inspiratory and end-expiratory CT were included from the Digital Lung Multi-center Study.The quantitative parameters of AT and emphysema were measured by both the voxel-based quantitative method and the conventional threshold method,respectively.All subjects underwent pulmonary function examination within 3 days after CT examination.For healthy volunteers,paired sample rank-sum test was used to compare the difference of quantitative parameters between voxel-based method and threshold method,Spearman rank correlation analysis was used to investigate the correlation between quantitative parameters of the two methods and pulmonary function.For COPD patients,the distribution and extent of AT and emphysema in patients with similar degree of pulmonary function (PFT) injury were observed.Results There were varying degrees of AT in the asymptomatic youth,with a median value of 5.70％ for the voxel-based method and with a median value of 7.96％ for the conventional threshold method,there was significant difference(Z=-4.015,P＜0.001).The correlation between AT and emphysema parameters of the voxel-based method and PFT parameters (r=-0.399 and-0.494,-0.335 and-0.439 separately,P＜0.05) were higher than that of the conventional threshold method,respectively (r=-0.357 and-0.453,-0.284 and-0.391,respectively;all P＜0.05).Furthermore,the voxel-based method can classify COPD patients with similar degree of pulmonary function injury into three subtypes:AT-dominant,emphysema-dominant,and mixed.Conclusions The voxel-based AT quantitative measurement method not only has high sensitivity and accuracy,but also provides imaging phenotype for the diagnosis of COPD and provides assistant decision-making for clinical management.

Objective:To explore the effect of Bushen Huoxue Decoction on ventricular remodeling and myocardial nuclear factor-kappaB (NF-κB) protein in rats with chronic heart failure.Methods:60 male SD rats were randomly divided into sham operation group (15 rats) and experimental group (45 rats). The rats of the experimental group was established CHF model by ligating the left anterior descending coronary artery combined with exhaustive swimming and starvation. Rats with chronic heart failure were randomly divided into model group, Bushen Huoxue group and lisinopril group.The Bushen Huoxue group was perfused with 15.75 g/(kg·d) Bushen Huoxue Decoction, the lisinopril group was perfused with 1.8 mg/(kg·d) of lisinopril suspension, and the sham operation group and model group were perfused with equal volume of distilled water. After 4 weeks of administration, the general mental state of rats was observed. The left ventricular internal systolic diameter (LVIDs) and internal diastolic diameter (LVIDd) were measured by cardiac color Doppler ultrasound, and the left ventricular ejection fraction (LVEF) and short axis shortening fraction (LVFS) were calculated. The expression of NF-κB protein in rat myocardium was detected by Western blot, and the morphology of left ventricular myocytes was observed by hematoxylin eosin staining.Results:Compared with the model group, the myocardial fibers of rats in Bushen Huoxue group and lisinopril group were arranged orderly, with few pyknosis, a small amount of inflammatory cell infiltration. Compared with the model group, the levels of LVIDs [(6.00±0.58)mm vs. (6.99±0.90)mm] and LVIDd [(3.96±0.51)mm vs. (5.14±0.57)mm] significantly decreased, LVEF [(54.48±6.75)% vs. (30.28±4.85)%] and LVFS [(33.86±4.27)% vs. (26.10±4.96)%] significantly increased, as well as the expression of myocardial NF-κB (1.06±0.10 vs. 1.58±0.29) protein significantly decreased ( P<0.05). Conclusion:Bushen Huoxue Decoction can resist ventricular remodeling,improve cardiac function and treat heart failure of CHF rats and the possible mechanism might be it could down-regulate myocardial NF-κB expression.