Administration, Inhalation ; Animals ; Disease Models, Animal ; Female ; Fluorine ; blood ; Fluorocarbons ; adverse effects ; pharmacokinetics ; Male ; Rabbits ; Rats

Objective To establish a desirable quantitative assay system to evaluate the correlation between surgical manipulation and micrometastasis in primary esophageal cancer patients.Methods 118 pe- ripheral blood samples from 40 esophageal cancer patients undergoing radical resection were collected before surgery(B-1),immediately after surgery(B0)and at the third day postoperatively(B+3).Based on real-time quantitative reverse transcription-PCR,carcinoembryonic antigen(CEA)mRNA gene was used in the detec- tion.12 patients with benign tumor and 13 healthy volunteers were collected as negative control groups.Re- sults The median of CEA mRNA copies at B-1,B0 and B+3 were 1592,13 314 and 6221 copies/ml blood, respectively.CEA mRNA expression levels were found to be significantly higher at both B0 and B+3 than that of B-1(P=0.0001 and 0.0209,respectively).No significant difference was found between B0 and B+3(P= 0.4396).Conclusion Surgical manipulation on esophageal cancer patients increases the probability of mi- crometastasis.Therefore,adjuvant therapy is needed during perioperative stage.

Emergencies ; Environmental Exposure ; standards ; Humans ; Maximum Allowable Concentration

Gas Chromatography-Mass Spectrometry ; Solvents ; analysis ; chemistry ; Volatile Organic Compounds ; analysis ; chemistry

Objective To investigate the effect of fluoride on the expression of a subunit of calcium/calmodulin-dependent protein kinase- Ⅱ (α-CaMK Ⅱ ) at both mRNA and protein levels in human neuroblastoma cells were cultured in DMEM with final concentrations of NaF 0(control) ,0.05,0.50,2.00,5.00 mmol/L, respectively, for 48 hours. Then quantitative RT-PCR and Western blot were performed to detect the expression level of α-CaMK Ⅱ P1 (postnatal day 1) pups together with their mothers were randomly divided into three groups. Lactating rats were given drinking water containing NaF at concentrations 0(control) ,2,3 mmol/L. And pups were exposed to NaF through milk. In each group, 8 pups were sacrificed on day 14 after birth. In post-weaning period, another 8 pups in each group were given drinking water with the same dose of fluoride as their mother's 21 day after birth. After then, these pups were killed on day 28, and hippocampus was dissected immediately and Western blot was conducted mRNA and protein levels were decreased. When NaF concentrations were 0,0.05,0.50,2.00,5.00 mmol/L, the mRNA relative ratios of α-CaMKⅡ in SY5Y cells were 1.00 ± 0.00,0.77 ± 0.18,0.40 ± 0.11,0.22 ± 0.06 and 0.15 ± 0.03, and protein levels of α-CaMK Ⅱ were 100.00 ± 0.00,76.17 ± 2.08,59.16 ± 2.12,48.52 ± 2.71 and 43.51 ± 2.57, any mmol/L group, hippocampus α-CaMK Ⅱ protein levels on day 14 and 28(75.02 ± 2.88,73.83 ± 3.88 and 81.00 ± 2.54,45.70 ± 2.34) were significantly lower than that of control groups(100.00 ± 0.00,100.00 ± 0.00, all P < 0.01). In 3 mmol/L group, hippocampus α-CaMKⅡ protein level on day 28 was lower than that of 2 mmol/L group (P < 0.01). Conclusion Fluoride can decrease mRNA and protein levels of α-CaMK Ⅱ in nerve cells and hippocampus, which may be one of the mechanisms of learning and memory impairment by fluorosis.

Objective To investigate the proliferation and identification of dendritic cells(DC)de- rived from peripheral blood of patients with bladder cancer in vitro.Methods The mononuclear cells were prepared from peripheral blood of patients with bladder cancer by Ficoll-Hypaque centrifugation method,and were induced by the recombinant cytokines hGM-CSF(50 ng/ml),hlL-4(10 ng/ml)and hTNF-?(50 ng/ ml)for 2 weeks.The growth and morphology of DC were observed through the phase contrast or electron mi- croscope,and their pheuotypes were determined by flow cytometry.The capacity of DC to activate T cell-de- pendent anti-tumor immune responses was tested by MTT method.Results The DC cultured in vitro turned into suspensive growth from adhesive situation on the 6th day,then the number of DC increased con- tinuously and the cells showed the irregular morphologic appearance of DC with veiled edges on the 8th day. Flow cytometry showed that the mature DC expressed high levels of specific markers such as CD_(1a),CD_(83), CD_(86)and HLA-DR.T cells activated by DC showed strong cytotoxicity to bladder cancer cell line BIU87 with a killing rate of(48.8?3.7)%,while the killing rate of T cells which were not activated by DC was(25.7?1.5)%;the difference of the rate between them was significant(P＜0.01). Conclusions The DC can be cultured from peripheral blood of patients with bladder cancer by induction of rhGM-CSF,rhIL-4 and hT- NF-?in vitro.This may lay an experimental foundation for further research on DC vaccine.

Objective To investigate the method of directed differentiation dendritic cells from embryonic stem cells(ESC) and to amplify high purity DCS in vitro for immunity therapy.Methods E14 ESC line were generated ESC-derived dendritic cells(ES-DC) in complete medium further supplemented with granulocyte-macrophage colony-stimulating factor(GM-CSF) and interleukin-3(IL-3).ES-DCs was used flow cytometry to determine CD11c,CD80,CD86,MHC-Ⅱ cell surface phenotype. Lipopolysaccharide (LPS) were added to induce the ES-DCs matured. The matured ES-DCs was harvested 24 hours later to be identified with morphology, transmission electron microscopy, analyzed by flow cytometry and compared with the immatured ES-DCs phenotype. The antigen presenting was evaluated by mixed lymphocyte responses.Results The ES-DC had obviously dendritic processes under scanning electron microscope . The immature DCs express low level of CD11c(4.33±0.23)%,CD80 (7.62±0.19) %, CD86 (4.77±1.22) % and MHC-Ⅱ (9.68±0.15) %, but the mature DCs express higher lerve of CD11c(47.36±2.68)%,CD80 (74.4±1.47) %, CD86 (29.77±2.00) % and MHC-Ⅱ (87.56±2.75) %. MLR showed that ES-DCs could effectively stimulate lymphocyte to proliferate.Conclusion These results provide evidence that DCs can be generated from E14 ESC with GM-CSF and IL-3, express high level of CD11c,CD80, CD86, MHC-Ⅱ and can effectively stimulate lymphocyte to proliferate. ES cells may become new origin for DCs which provided the immunotherapy.

Objective To explore whether human umbilical venous endothelial cells could be used as feeder layer to support the growth of embryonic stem cells (ESC) and keep ESC undifferentiated.Methods The venous vessels of umbilical cord obtained from healthy puerperal were perfused with collagenase.The isolated endothelial cells went through primary culture and passages for expansion.Factor Ⅷ antigens determination was implemented.Endothelial cells with good growth and 3 or above passages were treated with mitomycin-C(10 mg/L) and prepared as feeder layer,on which E14.1 ESC was transplanted for subculturing to observe the morphological characterization and determine ESC alkaline psphatase (AKP) activity and the expression of stem cell marker Oct-4.Severe combined immune deficiency(SCID) mouse in vivo terotoma formation experiment was performed to identify its pluripotent properties.Results Human umbilical vein-derived endothelial cells grew well in culturing in vitro and regenerate in large numbers.The endothelial cells maintained normal cellular morphological and biological characterization after 10 passages.The cells stopped proliferating after being treated with mitomycin-C,but its activity and morphological properties were well-maintained with 24 hours,which was a fundamental property of serving as feeder layer.E14.1 ESC remained undifferentiated in human umbilical venous endothelial cells after 3-8 passages,the cells grew in colony and showed high expression of AKP and stem cell Oct-4.In vivo pluripotency experiment showed that 6 weeks after being transplanted to SCID mice E14.1 ESC of 6 and 10 passages in endothelial cells both could form teratoma containing 3 layers of tissue cells.Conclusions Human umbilical venous endothelial cell serve as a convenient feeder layer cell with rich sources.It can effectively support ESC growth and heterogenous and prevent the heterogeneous protein pollution and pathogenic microorganisms caused by animal cell feeder layers,thus solve the problem of biological safety of ESC clinical application.

Objective To study CD64 expression in neutrophilic granulocyte and its clinical effect in neonatal infection disease. Methods CD64 was detected among 59 neonatal patients(septicemia group 34 patients, local infection group 25 patients)hospitalized in our neonatal department diagnosed as neonatal infection disease in 48 h after hospitalized,2 weeks after therapy, then the results were compared with 27 patients as non - infection disease during the same period. Results CD64 in septicemia group was (6156. 21?3643. 32) molecula per cell,in local infection group was (2176.19 ? 946. 32)molecula per cell, in non- infection group was (2176. 19 ? 946. 32) molecula per cell.There were significant differences among three groups (all P0.05). Conclusions CD64 expression increases in bacterium infection disease. It is more obvious in widespread infection desease.and it can be the mark in early diagnosis of neonatal infection disease.

To explore the effect of tripterygium glycosides on the level of peripheral blood cell factors of Graves ophthalmopathy (GO). In the study, 64 patients of GO in moderate-severe acute stage were selected, and randomly divided into the treatment group (32 cases) and the control group (32 cases). Both of the two groups were provided with basic treatment. The control group was added with prednisone(0. 75 mg kg-1 d-1 ), which gradually reduced (by 5-10 mg week-1 )to the minimum dose of 5 mg d-1. The treatment group was treated with 20 mg tripterygium glycosides, three times a day. One therapy course is three months. The levels of peripheral blood cells(TNF-alpha , IL-2, IL-10, IFN-gamma)of the two groups before and after the treatment and the clinical efficacy were observed. The study indicated that, before the treatment, TNF-alpha, IL-2, IFN-gamma in both groups were significantly higher than that in the health group, but with IL-10 notably lower than the healthy group. After the treatment, TNF-a, IL-2, IFN-gamma in the treatment group significantly decreased, but with IL-10 significantly increasing (P <0. 01). After the treatment, the two groups showed significant difference (P <0. 01). The total clinical efficacy in the treatment group was 88. 10% , and that in the control group was 57. 14% (P <0. 01). After the treatment, the two groups showed significant changes in the exophthalmos degree (P < 0. 01). The results showed that the level of peripheral blood cells (TNF-alpha,IL-2, IL-10, IFN-gamma)of GO patients was positively correlated with the severity of ocular disease. The combined therapy of tripterygium glycosides and methimazole show such advantages as low side effect and high clin-
Adult ; Cytokines ; blood ; Female ; Glycosides ; pharmacology ; therapeutic use ; Graves Ophthalmopathy ; blood ; drug therapy ; Humans ; Male ; Tripterygium ; chemistry