Objective To investigate the role of anti apoptosis gene bcl 2 in the survival of cultured uveal melanoma UM cells. Methods Antisense oligodeoxynucleotides (AS ODN) bcl 2 were delivered with cationic lipid to primary cultured UM cells. The inhibitory effect of AS ODN bcl 2 on proliferation of UM cells was examined by 3, 4,5 Dimethyliazol 2,5 diphenyl tetrazolium bromide (MTT) method. Using DNA ladder to determine if the UM cells had been apoptotic. Bcl 2 expression was detected by RT PCR and Westernblot technics. Results The effect of AS ODN bcl 2 in inhibiting the proliferation of cultured UM cells had opposite relation to dosage. It down regulated the mRNA and protein level of bcl 2 gene, and the sense ODN didn′t have this effect. Conclusion AS ODN bcl 2 can down regulate bcl 2 expression, inhibits UM cells proliferation and induces apoptosis.

Aiming at motion artifacts and off-resonance artifacts in multi-shot diffusion magnetic resonance imaging (MRI), we proposed a joint correction method in this paper to correct the two kinds of artifacts simultaneously without additional acquisition of navigation data and field map. We utilized the proposed method using multi-shot variable density spiral sequence to acquire MRI data and used auto-focusing technique for image deblurring. We also used direct method or iterative method to correct motion induced phase errors in the process of deblurring. In vivo MRI experiments demonstrated that the proposed method could effectively suppress motion artifacts and off-resonance artifacts and achieve images with fine structures. In addition, the scan time was not increased in applying the proposed method.
Artifacts ; Diffusion Magnetic Resonance Imaging ; Humans ; Image Processing, Computer-Assisted ; Motion ; Phantoms, Imaging ; Vibration

Adolescent ; Adult ; Boron ; adverse effects ; Humans ; Male ; Occupational Exposure ; Sperm Count ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects

BackgroundResearches demonstrated that the levels of soluble CD44 (sCD44)molecule in aqueous is significantly higher in primary open-angle glaucomous(POAG) eye than normal eye,but how the sCD44 would affect the expression of apoptosis protein in trabecular meshwork cells is below understanding.Objective The present study was to investigate the effect of sCD44 on the expression of regulatory proteins bcl-2 associated death factor bad in trabecular meshwork cells in the patients with POAG.MethodsHuman scleral tissue with trabecular meshwork were obtained from POAG patients during the surgery.The trabecular meshwork cells were primarily cultured by explant culture method and identified by immunochemistry.The third generation of cells were incubated with free-serum DMEM/F12 medium added differnt dosages of sCD44 (0,1,5,10,25,50 mg/L) for 48 hours.The expression of bad protein in cultured cells was detected using cell counting kit-8 (CCK-8) as the absorbance values at 490 nm(A,90 value),and the bad protein level in cultured cells was assayed by ELISA.ResultsThe cultured cells showed the positive response for laminin ( LM ),neuron specific enolase ( NSE ),fibronectin ( FN ) monoclonal antibodies.The CCK-8 assay showed that the A490 values of the trabecular meshwork cells in 0,1,5,10,25,50 μg/L of sCD44 groups were 0.2460±0.0019,0.1874±0.0015,0.1570±0.0016,0.1302±0.0019,0.1084±0.0018,0.0940±0.0020 respectively with a statistically significant difference among the 6 groups( F =14.922,P =0.000 ),and the A490 values in various dosages of sCD44 groups were significantly lower than the 0 μg/L sCD44 group (P=0.013,0.008,0.011,0.005,0.004).The ELISA assay showed that bad protein levels in 0,1,5,10,25,50 μg/L of sCD44 groups were ( 114.8461 ± 2.9560 ),( 137.8270 ± 2.4259 ),( 161.4194 ± 3.7381 ),( 170.9453 ± 3.2006 ),( 221.2252 ±4.3738 ),( 324.6167±4.4220) ng/L,showing a total difference among them ( F =16.610,P =0.000 ),and the bad protein levels in various dosages of sCD44 groups were significantly lower than the 0 μg/L sCD44 group( P =0.017,0.013,0.008,0.007,0.006).ConclusionssCD44 can contribute to the apoptosis of the trabecular meshwork cells in patients with POAG in certain dose range by regulating the apoptosis regulatory proteins bcl-2 associated death factor bad.

Background Primary open angle glaucoma (POAG) is a major blindness-causing disease,characterized by elevated intraocular pressure due to an insufficient outflow of aqueous humor. The trabecular meshwork lining the aqueous outflow pathway modulates the aqueous outflow facility. To study the biological characteristics of the trabecular meshwork cells has important significance. Objective This study was to culture the trabecular cells from primary open-angle glaucomatous eye (POAG) and study the biologic characteristics of passaged cells. Methods The deep scleral tissue with trabecular meshwork was obtained during the trabeculectomy from 8 eyes with POAG. The trabecular cells were primarily cultured and passaged in vitro. The generation 3 cells were identified by immunochemistry with the laminin (LM), fibronectin (FN) and neuron specific endolase (NSE)monoclonal antibodies. The ultrastructure was examined to observe the biological characteristics of the cells under the transmission electronic microscope. The experimental results were compared among POAG group, normal control group and blank control group. Results The primarily cultured POAG trabecular cells migrated from the edge of tissue mass about 10 days. The cells of generation 3 presented the logarithmic phase in the first 4 days and fused in the 7th day. FN,LM and NSE were positively expressed in the generated cells in POAG group and normal control group rather than blank control group. The MOD values of the generation 3 cells for FN in POAG group and normal control group were 0. 35 ± 0.06 and 0. 26 ± 0. 01, and those for LM were 0. 34 ± 0. 03 and 0. 25 ± 0. 02 respectively, showing statistically significant difference between these two groups ( FN: t = 14. 446, P＜0.001; LM: t = 9. 346, P＜0. 001 ). The microvilli, cytolysosome and phagocytic vesicle were obviously decreased in the trabcular cells of POAG group compared with normal control group under the transmission electron microscope. Conclusion The trabecular meshwork cells from POAG can be successfully cultured and passaged in vitro. It provides a cytology basis for further glaucoma research.

Objective To evaluate the clinical efficacy of oropharyngeal decontamination combined with gastrointestinal excitomotor in preventing hospital-acquired lower respiratory tract infection(HALRTI)in elderly inpatients. Methods Totally 110 elderly inpatients with high risks of HALRTI were randomized into intervention group and control group. Patients of the intervention group were administrated by cetylpyridinium chloride gargle twice daily for gargling(20 mL for each), combined with moshabili three times daily orally(5 mg for each). Exact Sig. l-sided was performed to compare the occurence of HALRTI,and Spearman test was performed to evaluate the correlation between positive rate of the phyarynx swabs and the hospitalized time. SPSS 12.0 software was used for statistical analyzing. Results Patients suffered from HALRTI were 1 case(2.00%)in intervention group and 7 cases(13.73%)in control group respectively,and the difference was of statistical significance(P ＜ 0.05). As hospitalized time went on, the positive rate of the phyarynx swabs microbial culture in the control group increased(r = 0.450, P ＜ 0.05), while the intervention group showed no significant change during the whole intervention time(r = -0.023, P ＞0.05). Conclusion The combined intervention of oropharyngeal decontamination with the gastrointestinal excitomotor may be beneficial to elderly inpatients with high risk of HALRTI in the prevention of HALRTI.

Child ; Humans ; Liver Failure ; therapy ; Liver Transplantation ; Male ; Plasma Exchange

ObjectiveTo investigate the effects of Tai Chi exercise on somatic function of middle-aged females.Methods60 middle-aged females had a 16-week Taichi quan exercise and changes of blood pressure, resting heart rate and vital capacity were tested.ResultsAfter Tai Chi exercise, 60 women had results of resting heart rate and blood pressure decreased, and vital capacity increased obviously (P<0.05～0.01).ConclusionTai Chi exercise maybe an effect sport manner to improve the somatic function of middle-aged females.

OBJECTIVETo observe the effect of nourishing Yin, strengthening Qi and activating blood decoction on Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression.

METHODThirty-two NOD mice were randomly divided into the model group, the traditional Chinese medicine group (TCM group, orally given 0.4 mL nourishing Yin, strengthening Qi and activating blood decoction as per 100 g x kg(-1) everyday), the hydroxychloroquine group (given 0.4 mL hydroxychloroquine as per 60 mg x kg(-1) everyday), the traditional Chinese medicine and western medicine group (TCM WM group, given nourishing Yin, Strengthening Qi and activating blood decoction 50 g x kg(-1) and hydroxychloroquine 60 mg x kg(-1), 0.4 mL everyday), with eight mice in each group. Eight Balb/C mice were selected as the normal control group (normal group). All of mice were killed after eight weeks, and their submaxillary glands were dissected. The expression levels of Fas/FasL were examined by immunohistochemical method, and the FasL mRNA was detected by RT-PCR.

RESULTThe expression levels of Fas/FasL in salivary glands of the model group were higher than that of other groups (P < 0.05). The expression level of FasL of the normal group was much lower than that in the hydroxychloroquine group (P < 0.05). The relative expression level of Fas mRNA in salivary glands of the model group was higher than that in other groups, but the control group was notably lower than other groups (P < 0.05). The expression level of FasL mRNA in salivary glands of the model group was higher than that in TCM and TCM WM groups (P < 0.05). But the expression level in TCM WM group was notably lower than the hydroxychloroquine group (P < 0.05).

CONCLUSIONThe nourishing Yin, strengthening Qi and activating blood decoction could down-regulate the expression level of Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression, and had a better efficacy after being combined with hydroxychloroquine. The nourishing Yin, strengthening Qi and activating blood decoction might treat the Sjogren's Syndrome by reducing apoptosis which is regulated by Fas/FasL

Animals ; Fas Ligand Protein ; genetics ; Female ; Gene Expression Regulation ; Medicine, Chinese Traditional ; methods ; Mice ; Mice, Inbred NOD ; Qi ; RNA, Messenger ; genetics ; metabolism ; Salivary Glands ; metabolism ; Sjogren's Syndrome ; blood ; genetics ; therapy ; Yin-Yang ; fas Receptor ; genetics

Objective To identify the dynamic influence of antibiotics use on T helper cell(Th) 1/Th2 cytokine balance in neonatal rats in their early life.Methods Twenty-four newborn SD rats were randomly divided into 3 groups including 2dCef group,7dCef group and a control group (n =8).SD rats at 2 or 7 days of age were injected Cefotaxime sodium intraperitoneally for 3 consecutive days.The control group was injected the same amount of phosphate buffer saline.Serum and bronchoaveolar lavage fluid (BALF) samples were collected on 35 days old and diffe-rential cell count in BALF was done.Enzyme linked immunosorbent was used to detect the concentration of interleukin(IL)-12,interferon-γ(IFN-γ),IL-4 and IL-5 in serum and BALF,and real-time quantitative PCR was used to detect mRNA expression of those cytokines in lung tissues.Results The number of total cells,neutrophils,and lymphocytes in Cefotaxime treated on 2nd day after birth were significantly higher than those in the controls and 7th day group (F =93.78,15.84,7.36,all P ＜0.05).For Thl cytokines detection:the IL-12 secretion in BALF and serum and INF -γsecretion in BALF of the 2 antibiotic groups were lower than those in control group (F =26.53,12.95,47.18,all P ＜ 0.05),the decrease in 2dCef group were more obviously than 7dCef group [(127.72 ± 12.35) ng/L vs (198.15 ± 18.93) ng/L,(105.91±13.61) ng/Lvs (133.63 ±13.82) ng/L,(147.23 ±24.10) ng/Lvs (178.52±12.46) ng/L,all P ＜0.05].The mRNA expression of IL-12 in antibiotic groups were lower than those in control group (F =33.60,P ＜0.05),and the decrease in 2dCef group was more obvious than that in 7 dCef group (0.073 ± 0.023 vs 0.093 ± 0.028,P ＜0.05).For the Th2 cytokines:the IL-4 secretion concentration and mRNA expression in the 2 antibiotic groups were higher than those in the control group (BALF,serum and lung) (F =64.26,13.79,11.54,all P ＜ 0.05),the increase in 2dCef group was more obvious than that in 7dCef group[(103.65 ± 7.68) ng/L vs (82.04 ± 8.77) ng/L,(87.81 ± 4.72) ng/L vs (69.69 ± 7.79) ng/L,(0.067 ± 0.036) ng/L vs (0.051 ± 0.055),all P ＜ 0.05];however,no significant differences of IL-5 either mRNA or cytokine secretion being detected among these 3 groups (F =0.50,0.75,0.08,all P ＞ 0.05).Conclusions Antibiotic use during early time of neonate rats alters postnatal immune cells distribution in the lung and promotes the shift in cytokine balance towards a Th2 profile,and the earlier the antibiotic use,the more serious the impact.