Biomedical Research ; Humans ; MicroRNAs ; Neoplasm Invasiveness ; genetics ; Neoplasm Metastasis ; genetics

Objective To explore the changes of intereferon-?(IFN-?)and interleukin-4(IL-4)in peripheral blood of children with mycoplasma pneumoniae(MP) encephalitis at the acute phase.Methods The peripheral blood concentrations of IFN-? and IL-4 were measured by enzyme-linked immunosorbent assay(ELISA) in 24 cases of children with MP encephalitis at the acute stage.The samples from 24 cases of healthy children were control group.Twenty-four children with MP encephalitis were intravenous drip with azithromycin,at the same time,10 cases received hexadecadrol and 15 cases received gamma globulin.Results The serum concentrations of IFN-? and IL-4 in the mycoplasmal encephalitis group were(98.56?12.93) and(45.55?17.58) ng/L,respectively,which were significantly higher than those in control group [(85.35?6.91) and(26.78?9.89) ng/L] respectively(Pa

Objective To establish a rapid method for the isolation of murine peritoneal macrophages. Methods Abdominal lavage was performed with NS and the peritoneal macrophages was purified from the lavage,which was later transferred into high glucose DMEM cul-ture medium. Cell viability was measured via the typan blue staining. Purity was observed via Wright's stain. Results High purity macropha-ges with typical morphology were obtained. Conclusion A simple and realistic method was set up for the isolation of murine peritoneal mac-rophages.

Objective:To study the clinical efficacy of acupuncture for ischemic stroke and its effect on ET-1 levels in IS cases.Method:The 63 cases were randomized into a treatment group (31 cases),receiving acupuncture and routine method,and control group (32 cases),receiving routine method alone.Seven days constitute a course of treatment,a 2-day interval between two courses.The clinical efficacy and ET-1 level in two groups were compared after four weeks.Result:The total effective rate in the treatment group and control group were 96.8% and 75% respectively,with a statistical significance (P＜0.05);the ET-1 level in blood plasma also with a statistical significance after the treatment (P＜0.05);the pre-treatment and post-treatment ET-1 level in the treatment group showed a statistical significance (P＜0.01),whereas the control group didn't.Conclusion:Acupuncture is effective for ischemic stroke and can lower the ET-1 level in ischemic stroke cases.

Objective To measure the expression of activin receptor-like kinases 1(ALK1)in dermal fibroblasts from patients with systemic scleroderma(SSc)and to estimate its role in the production of fibronectin and plasminogen activator inhibitor-1(PAI-1).Methods Dermal fibroblasts were isolated from the lesions of 12 patients with SSc as well as the normal skin of 14 healthy controls,and subjected to a primary culture.The third-passage fibroblasts were used in the next experiment.Western blot and indirect immunofluorescence technique were utilized to quantify the expression of ALK1.A specific siRNA targeting ALK1 was designed,constructed,and transiently transfected into the control dermal fibroblasts,which were then classified into 2 groups to be cultured with or without the presence of transforming growth factor(TGF)-β1 for 72 hours followed by the detection of fibronectin and PAI-1 expression with Western blot.Results As Western blot and direct immunofluorescence technique showed,both control and SSc fibroblasts showed an expression of ALK1 in the cytoplasm and membrane,and the expression intensity of ALK1 in SSc fibroblasts was significantly higher than that in the control fibroblasts(1.97 ± 0.05 vs.1.12 ± 0.03,t =50.96,P ＜ 0.05).The expression of ALK1,fibronectin and PAI-1 was decreased by 90％,58％ and 31％ respectively in specific siRNA-transfected SSc fibroblasts compared with the control siRNA-transfected fibroblasts.TGFβ1 significantly increased the expression of ALK1,fibronectin and PAI-1 in the control siRNA-transfected fibroblasts,but the increase was markedly inhibited by the siRNA-targeting ALK1.Conlusion TGFβ1 can promote the production of fibronectin and PAI-1 via ALK1 in fibroblasts,and ALK1 may be involved in the development of sclerosis in SSc.

Objective To investigate the effect of chemotherapy regimen of rituxmab combined with CHOP (R-CHOP) on the survival of patients with diffuse large B cell lymphoma (DLBCL). Methods One hundred and fifty-six cases of DLBCL diagnosed according to the WHO 2008 classification were collected from the haematopathological laboratory, the department of pathology, and Beijing University Health Science Center. Standard two-step method of immunohistochemical staining with Envision was used to assess the expression of CD10, MUM-1, bcl-6, and bcl-2. The different classification models were made according to the immuaohistochemical staining results. Hans algorithm classifies the patients into two subgroups originating from germinal center B cell-like cell (GCB) and non-germinal center B cell-like cell (non-GCB), and Muris model were classfied the DLBCL patients into the good-survival groupl and the poor-survival group2. Thirty patients with treatment of R-CHOP were set as study group and the other 126 patients without Retuxmab were defied as control group. The data were analyzed with X2 test, log-linear model and Life Table survival analysis by the SAS 8.2 statistical package. Results The 3-year survival rate of the study group was 78.3 %, but was 53.4 % in the control group. The over-all survival of the study group was obviously better than the control group with the significant difference (P <0.05). Hans algorithm showed no implication of survival for any group. The survival of different groups in Muris model has no difference in study group but was obvious in control group. The expression of bcl-2 protein has no association with survival in study group but acted as a worse implication of survival in control group. Conclusion R-CHOP chemotherapy regimen could improve the remission rate and over-all survival of DLBCL. Rituxmab could weaken the effect of bcl-2 expression in the prognosis, and the implication of survival by Muris model has diminished.

Objective:To investigate the effects and mechanism of β-carotene on inflammatory factors (IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells.Methods:Firstly,RAW264.7 cells of being induced by 4 (5 μg/ml)for 24 h were treated with different concentration of β-carotene (20,40,80,160 pmol/L)for 3 h.The cells viability was measured by MTIT,the mRNA relative expression of IL-1 β,IL-6,TNF-cα was detected by fluorescence quantitative PCR,the secretion capacity of IL-1 β,IL-6,TNF-α was detected by ELISA and the protein relative expression of NF-κB p65 protein was measured by Western blot.Secondly,RAW264.7 cells were induced by LPS(5 μg/ml) and different concentration of PDTC(1,5,10 μg/ml)for 24 h,NF-κB p65 protein was measured by Western blot and inflammatory factors were detected by fluorescence quantitative PCR and ELISA.Finally,compared the changes in the relative expression of inflammatory factors and NF-κB p65 protein between LPS+PDTC group and LPS+PDTC + β-carotene group.Results:Compared with the LPS-induced group,β-carotene could increase the cell viability of LPS-induced RAW264.7 cells and inhibied the relative expression of inflammatory factors and NF-κB p65 protein.Inhibited the relative expression of NF-κB p65 protein could reduce the relative expression of inflammatory factors.Compared with the LPS+PDTC group,LPS +PDTC + β-carotene group could inhibit the relative expression of inflammatory factors significantly (P＜0.05).But,there was little difference about the relative expression of NF-κB p65 protein between this two groups.Conclusion:β-carotene inhibits the relative expression of inflammatory factors(IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells through inhibition of NF-κB p65 protein in NF-κB pathway,this pathway isn't unique.

Background Choroidal melanoma(CM)is a common form of primary ocular cancer in adults.It is reported that indomethacin has inhibitory effect on many tumor in vitro and in vivo,but whether it can inhibit the growth of CM has not been published. Objective This study was to investigate the anti-tumor activity of indomethacin on the growth of human CM OCM-1 cell xenografts in nude mice. Methods OCM-1 cells were subcutaneously implanted on 24 SPF female BALB/C.nu/nu nude mice to establish ectopic models of human CM.The nude mice with the tumor 5 mm were randomly divided into 4 groups:untreated group,normal saline solution(NS) group,indomethacin 1 ms/kg group,indomethacin 2 ms/kg group.The 1 mS/kg or 2 ms/kg indomethacin was intraperitoneally injected for 14 consecutive days in indomethacin 1 ms/kg group and indomethacin 2 me/kg group respectively.and 0.2 ml of 2%NS-DMSO was used at a same way in the NS group.No any agent was used as the untreated group.The volume and weight of implanted tumor as well as inhibitory rates of indomethaein on tumor were calculated.The expression of ki67 and survivin proteins were measured with immunohistochemistry,and the expression of survivin mRNA in CM was assessed by RT-PCR. ResuIts The tumor of indomethacin treatment group was reduced in volume and weight with a significant difference between treatment group and control group as well as indomethacin 1 ms/ks group and indomethacin 2 ms/kg group(P＜0.05).The inhibitory rate of indomethacin 1 ms/kg and 2 ms/kg for tumor was 22.86%,48.00%respectively.The prolifiration index (PI)of ki67 in these 4 groups were (76.73±3.34)%,(73.30±2.95)%,(55.97±2.24)%,(32.87±2.91)%respectively,and significant difference was found in PI between indomethacin 2 mg/kg group and untreated group or NS group(P＜0.05),but there was not significant difference between indomethacin 1 mg/kg and 2 ms/kg group(P＞0.05).Compared with the control group,the indomethacin treatment groups showed the decreased expression of survivin protein and mRNA,and significant difference was found between indomethaein 2 ms/kg group and untreated group or NS group(P＜0.05),however,no significant difference was found between indomethacin 1 mg/kg and 2 mg/kg group(P＞0.05). Conclusion Indomethacin inhibits the growth of CM in nude mice through inhibiting the expression of survivin in the tumor and accelerating cell apoptosis and inhibiting tumor cell proliferation.

A rapid and convenient ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) method for analysis of ribavirin and its two main kinds of metabolites, 1,2,4-triazole-3-carboxamide (TCONH2 ) and 1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxylic acid (RTCOOH), in fresh eggs has been developed and validated. The sample was extracted with acetonitrile water (9 ∶ 1, V/ V), added N-hexane to move liposoluble substances, after centrifugation, the upper solvent was cleaned-up by C18 and GCB, then determined by UPLC-MS / MS coupled with Agilent ZORBAX SB-Aq column (100 mm×3. 0 mm, 1. 8 μm). Over the concentration range of 2. 0-200. 0 μg / L (0. 5-200. 0 μg / L and 5. 0-200. 0 μg / L) for ribavirin (TCONH2 and RTCOOH) in fresh egg matrix, the correlation coefficients (R2 ) of the calibration curves were above 0. 99, and the limits of detection (LODs) were 0. 54 μg / L (0. 09 and 1. 54 μg / L). The limits of quantitation (LOQs) were 1. 79, 0. 31 and 5. 13 μg / L, respectively. At three spiked concentration levels of 5. 0, 10. 0 and 50. 0 μg / L, the recoveries of ribavirin and RTCOOH ranged from 96. 1% to 99. 6%and 42. 9% to 58. 3% . And at three spiked concentration levels of 0. 5, 2. 0 and 5. 0 μg / L, the recoveries of TCONH2 ranged from 75. 9% to 106. 7% . The results of the determination in various real samples showed that the method is simple, accurate, rapid and suitable determination of ribavirin and its two main kinds of metabolites in fresh eggs.