Objective: To investigate the effects of artemisinic acid on proliferation of K562 cells and its mechanism in vitro.Methods: MTT assay was used to detect the effects of artemisinic acid on proliferation of K562 cells.The flow cytometry was used to analyze the distribution of cell cycle.Morphological changes of cells were observed by electron microscopy.Results: The proliferation of K562 cells was inhibited by intervention of artemisinic acid significantly after 24 hours and 48 hours,and the effects were in concentration-dependent manner.The cell number in G0/G1 increased and apoptosis was induced after the treatment with artemisinic acid for 48 hours,the morphological characteristic of apoptosis was observed by electron microscopy.Conclusion: Artemisinic acid can inhibit the proliferation of K562 cells,which may be-due to induction of apoptosis and prohibit on of tumor cells from entering cell cycle.

OBJECTIVETo investigate the dynamic features of angiogenesis in residual tumors after high intensity focused ultrasound (HIFU),and to determine the temporal effect and mechanism of hypoxia inducible factor-2 alpha (HIF-2a) in the angiogenic process of residual tumors.

METHODSXenograft tumors of HepG2 cells were generated by subcutaneously inoculating athymic BALB/c nu/nu mice with the hepatoma cells.About 30 days after inoculation,all mice (except in the control group) were treated by HIFU and assigned randomly to the following 7 groups according to various time intervals post-treatment:1st,3rd,5th day and 1st,2nd,3rd,4th week when the residual tumor tissues were obtained from the experimental groups.Protein levels of HIF-2a and vascular growth factor A (VEGF-A) were quantified by immunohistochemistry and western blotting,and mRNA levels were measured by (real-time quantitative) qPCR. Microvascular density (MVD) was calculated by counting the CD31-positive vascular endothelial cells identified by means of an immunohistochemical staining method.

RESULTSCompared with results from the control group,the protein and mRNA levels of HIF-2a expression reached the highest level in the experimental mice at the 2nd week (P=0.000 and P < 0.01 respectively),and were decreased thereafter(3rd week and 4th week, P=0.000 and P < 0.05).VEGF-A expression in the residual tumor tissues group that received HIFU was significantly decreased,compared with the control group,at all time points uPto 1 week (all P=0.000 and P < 0.01),but the levels increased compared to controls in the 2nd through 4th week (all P=0.000, P < 0.05). Similar results were obtained for MVD.

CONCLUSIONHIFU treatment can inhibit angiogenesis in residual hepatoma tissues in the short-term (1 to 2 weeks post-treatment) in mice with hepatocellular carcinoma,but can promote angiogenesis overtime (2 to 4 weeks post-treatment); the angiogenic process may involve the HIF-2α/VEGFA pathway.

Animals ; Blotting, Western ; Carcinoma, Hepatocellular ; pathology ; Hep G2 Cells ; High-Intensity Focused Ultrasound Ablation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Immunohistochemistry ; Liver Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factor A ; metabolism

Objective To investigate the roles of interleukin(IL)-25,eosinophils in the occurrence of bronchial asthma.Methods Selected 30 cases of acute episode of asthma(asthma group),and ease a group of 23 cases of asthma treatment(remission group),20 cases of the normal control group.Using the method of gradient ultrasonic atomizing inhalation of Hyperosmotic saline collection of induced sputum specimens,by ELISA method for the determination IL-25 level in induced sputum,count of eosinophils,analysis of correlation between them.Results Group IL-25 in induced sputum of asthma and eosinophil levels[(313.12±75.64)ng/L vs(236.43±57.90)ng/L]were sig-nificandy higher than the control group[(0.386±0.267) × 109/L vs(0.005±0.002) × 109/L],and significantly higher than acute stage in remission stage[(268.63 + 40.19) ng/L,(0.120 + 0.016) × 109/L](P ＜ 0.05).Conclusion IL-25 has an important role in asthma development.It provides a new way of thinking for monitoring and treatment.

Objective To study the MRI of cervical spinal cord injury without fracture and dislocation and explore of the apparent diffusion coefficient (ADC) of injured site related to the severity of injury. Methods 46 patients with cervical spinal cord injury without fracture and dislocation and 20 healthy controls were scaned with routine MRI and diffusion weighted imaging. The ADC value of site of injury and grades of Frankel's classification were analyze with the correlation. Results There were 22 cases with spinal cord edema, 8 cases with intramedullary hemorrhage, 14 cases with edema and hemorrhage, 2 cases without abnormal finding. The ADC of controls and patients were (1.05±0.12)×10-3 mm2/s, (1.21±0.23)×10-3 mm2/s (t=0.704, P<0.05). The ADC values positively correlated with the grades of Frankel's classification (r=0.407, P<0.05). Conclusion MRI may help to find the cervical cord injury in those without fracture and dislocation, and the ADC may be resposible to the severity of injury.

AIM: To investigate the effect of epinephrine on LPS-induced pro-inflammatory mediators (TNF-?, NO and COX-2) and anti-inflammatory mediators (HO-1 and IL-10) production in murine macrophage RAW264.7 cells, and to determine whether these effect is due to the influence of epinephrine on NF-?B activation. METHODS: RAW264.7 cells were cultured in vitro with 10 ?g/L LPS in the absence or presence of epinephrine at variant concentrations (1, 5, 10, 50 ?mol/L) for 24 hours, then the supernatants was collected for measuring TNF-? and IL-10 by ELISA and Griess reagent was used to measure NO (NO_2-/NO_3-) concentration. At the same time point, cells were harvested and COX-2, HO-1 and I?B-? was detected by Western blotting. RESULTS: 10 ?g/L LPS significantly induced the production of TNF-?, NO (NO_2-/NO_3-), COX-2, HO-1 and IL-10. When epinephrine was added into the medium together with LPS, the pro-inflammatory mediators production was decreased in a dose-dependent manner, however, anti-inflammatory mediators HO-1 and IL-10 expression was enhanced by epinephrine. Epinephrine has no significant effect on I?B-? degradation in LPS-activated RAW264.7 cells. CONCLUSION: Epinephrine down-regulates LPS-induced pro-inflammatory mediator expression while promotes anti-inflammatory mediator production in murine macrophages. These effect seems to be independent of NF-?B activation.

OBJECTIVETo observe the effects of artemisiae annuae CQ-189 (AACQ-189) on proliferation of hNSC and HELF in vitro, and the main organ toxicity and the median lethal dose (LD50) of kunming mouse in vivo. The purpose is to approach that the toxicity and side effects of AACQ-189.

METHODUsing techniques of the colorimetric 5-diphenyl tetrazolium bromide (MTT) to detect the effects of AACQ-189 on proliferation of hNSC, and to detect the number of HELF survival by using techniques of trypan blue exclusion. To detect LD50 by tail vein injection in kunming mouse and using histomorphology method to observe the mouse main organ damage by AACQ-189.

RESULTAACQ-189 has low poisonous function on hNSC and HELF that our experimental concentration (3.125-12.5 mg x L(-1)) has already achieve an effective dose to inhibit the proliferation of Leukemia cells obviously. LD50 concentration of kunming mouse is 550 mg x kg(-1). Moreover, AACQ-189 has little effect to main organs at higher concentration.

CONCLUSIONAACQ-189 has low poisonous function, which is a natural anti-tumor drug and has a promising prospect for potential application. However we should do more research on its mechanism.

Animals ; Artemisia ; chemistry ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug-Related Side Effects and Adverse Reactions ; Female ; Fibroblasts ; cytology ; enzymology ; Humans ; Lung ; cytology ; Male ; Mice ; Plant Extracts ; adverse effects ; pharmacology ; Stem Cells ; drug effects

Objective To assess whether propofol call induce stable psychic dependence in the rats by self-administration experiment. Methods Twenty-four male SD rats 14 weeks old weighing 240一270 mg were studied. Anesthesia was performed with intraperitoneal injection of 3%sodium pentoharbitsl 40 ms/kg and atropine 03 mg/kg.A catheter wag inserted into the right external jugular vein. Penicillin(100 000 U)0.2 ml wag injected through the external jugular vein for anti-infection and heparin sodium(50U/ml)0.1 ml for anticoagulation. The self-administration experiment of 14 days was started after the 7 days of recovery. All the rats were randomly divided into 4 groups(n=6 each):contontrol group(C),propofol 0.56 mg/kg/l group(P1),propofol 1.00 mg/kg group(P2)and pmpofol 1.70 ms/kg group(P3).The experimental events were controlled by a computer with 50 times of the maximum injection per day.The times ofactive and inactive nose-poke response and times of drug iniection were recorded per day.Results Compared with group C and P1,the times of active nosepoke response and injections were significantly increased in group P2 and P3(P<0.01).The times of active nosepoke response and injections per day were significantly increased in group P3 than in group P2(P(0.01).There was no significant difference in the times of active nose-poke response and injections between group C and P1.There was no significant difference in inactive nose-poke resporme between the 4 groups.And the total daily doses of propofol injected in the last 3 days were significantly increased in a dose-dependent manner.Conclusion Propefol can induce the development of psychological dependence in rata and it is related to the dosage.

OBJECTIVETo study the changes in hypoxia-inducible factor (HIF1α, HIF2α) in the residual tumor cells in nude mice bearing hepatocellular carcinoma (HCC) following treatment with high-intensity focused ultrasound (HIFU).

METHODSThirty nude mice bearing human HCC received treatment with HIFU. At 1, 3, and 5 days and 1 and 2 weeks after the treatment, the mice were examined for pathological changes of the residual tumor with HE staining; SP immunohistochemistry, Western blotting and real-time quantitative PCR were used to detect the protein and mRNA expressions of HIF1α and HIF2α in the tumor.

RESULTSHE staining revealed the presence of residual tumor cells and large necrotic areas after the treatment. Immunohistochemistry showed a gradual increment of HIF1α protein and mRNA expressions after the treatment, reaching the peak level at 3 days (P<0.05) followed by progressive reduction at 5 days and 1 and 2 weeks. HIF2α expressions at either the protein or mRNA levels exhibited no significant changes within 3 days after the treatment (P>0.05) but increased significantly at 5 days and 1 and 2 weeks (P<0.05).

CONCLUSIONThe changes of HIF1α and HIF2α in the residual tumor after HIFU treatment in nude mice bearing HCC can be associated with tumor cell apoptosis and angiogenesis after the treatment.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; therapy ; Hep G2 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; therapy ; Mice ; Neoplasm, Residual ; metabolism ; pathology ; Ultrasonic Therapy ; methods

PURPOSE: The goal of this study was to explore the effects of hsa-let-7g on cell proliferation and apoptosis, and elucidate its role in lung cancer development.MATERIALS AND METHODS: The expression levels of has-let-7g and HOXB1 in tissues and cells were measured by qRT-PCR. An inhibitor of hsa-let-7g or one targeting a control messenger RNA were transfected into A549 and H1944 lung cancer cells, and the effects of hsa-let-7g dysregulation on cell viability and apoptosis were analyzed using CCK-8 and apoptosis detection assays. HOXB1 was confirmed as the target gene of hsa-let-7g, based on luciferase reporter assay results. The relationship between hsa-let-7g and HOXB1 was confirmed by co-transfection of inhibitors of hsa-let-7g and HOXB1 followed by Western blot, CCK-8, and apoptosis detection assays.RESULTS: We observed high expression of hsa-let-7g in lung cancer tissues compared to the corresponding normal tissues, and generally higher expression of hsa-let-7g in patients with advanced tumor classification. The results of CCK-8 and apoptosis detection experiments showed that the inhibition of hsa-let-7g significantly inhibited proliferation of A549 and H1944 cells, but also promoted apoptosis. HOXB1 is a specific target of hsa-let-7g, and downregulation of HOXB1 in lung cancer cells reversed the suppressive effects caused by knocking down hsa-let-7g.CONCLUSION: These data collectively suggest that the expression of hsa-let-7g inhibits lung cancer cells apoptosis and promotes proliferation by down-regulating HOXB1. The results from this study demonstrate the potential of hsa-let-7g/HOXB1 axis as a therapeutic target for the treatment of lung cancer.
Apoptosis ; Blotting, Western ; Cell Proliferation ; Cell Survival ; Classification ; Down-Regulation ; Humans ; Luciferases ; Lung Neoplasms ; Lung ; MicroRNAs ; RNA, Messenger ; Sincalide

China ; Humans ; Streptococcus pneumoniae