Using site-directed mutagenesis and DNA recombinant technology, GGC fragment was inserted into human IL-18 cDNA. The mutated IL-18 cDNA was constructed into plasmid pPIC9K, then transformed into Pichia pastoris GS115 and efficiently expressed. A Glycine residue was inserted into the recombinant IL-18 between Arg39 and Asp40 to form a RGD motif. The mutated IL-18 was termed IL-18-RGD. The protein was purified with Sephadex G-100. The cell culture of melanoma B16 showed that IL-18-RGD efficiently inhibited B16 tumor growth, IC50 = 8.10?mol/L, but the inhibitory effect of IL-18 was not detected. Both IL-18-RGD and IL-18 showed inhibitory activities on mice loaded B16 in vivo, the inhibitory efffct of IL-18-RGD was stronger than that of IL-18. The inhibitory activities of IL-18-RGD and IL-18 on bFGF induced angiogenesis on chorioallantoic membranes were detected. There was no differences between IL-18-RGD and wild IL-18 in the activity of inducing IFN-? in human PBMC.

Tricholoma giganteum is a famous species product of Jishou in the wes t of Hunan.It have not only special biological characteristics such as formatio n and ecological enviromment,but also delicious,fragrant with abundant nutrien t.It's mycelium can be isolated and growing well in medium,but its fruiting bo dy is difficult to be formed.

Acute Disease ; Adult ; Emergency Nursing ; First Aid ; Humans ; Male ; Nitriles ; poisoning ; Young Adult

Objective To investiagte the serum adiponectin concentration in preeclampsia and its relationship with serum leptin and soluble leptin receptor levels.Method The level of adiponectin,leptin and soluble leptin receptor in serum were measured by enzyme-linked immunosorbent assay(ELISA)in 38 patients with preeclampsia and 42 patients as control.The relationship of free leptin index(leptin/soluble leptin receptor) to preeclampsia was analyzed.Results There were no significant differences in maternal age,gestational age and body mass index(BMI)between two groups.But the gestational age and birth weight were significantly lower in preeclampsia than in control.The patients with preeclampsia had significantly higher levels of serum adiponectin, leptin and free leptin index(1691.7?g/ml,37.5 ng/ml and 0.95 respectively)than the control(689.4?g/ml, 19.3 ag/ml and 0.49,respectively).But there was no significant difference in serum level of soluble leptin receptor between the groups(35.0 ng/ml vs 42.2 ng/ml).Serum adiponectin was not significantly correlated with the level of leptin,soluble leptin receptor and free leptin index.Area of serum adiponectin,leptin and free leptin index in preeclampsia under the ROC curve were less than 0.5.Conclusions The patients with preeclampsia have paradoxical higher serum levels of adiponecfin and more bioavailability of leptin,suggesting these may be important factom of this complication.

Objective:To investigate the expression of sphingosine kinase 1 (SphK1) and its clinical significance in breast invasive ductal carcinoma ( BIDC).Methods: We detected SphK1 expression in 58 samples of surgically resected paired BIDC and normal tumor-adjacent tissues samples by using immunohistochemistry.The correlation between SphK 1 expression and clinicopathologic features was analyzed.Results: The positive expression rate of SphK 1 in BIDC tissues was 69.0% ( 40/58 ) , while its positive expression rate in normal tumor-adjacent was 17.2% (10/58),the difference was statistical significance (χ2=31.636,P=0.000). Clinicopathological evaluation suggested that SphK 1 positive expression was associated with ER negative (χ2=4.392,P=0.036),PR negative (χ2=7.920 , P=0.005 ) , lymph node metastasis (χ2 =5.033 , P=0.025 ) and tumor stage (χ2 =7.117 , P=0.008 ) . Conclusion:The high-expression of SphK1 is correlated with the poor clinicopathological features in BIDC ,suggesting SphK1 may play a key role in development and progression of BIDC.

Objective To explore the impacts of over-expression of microRNA-7 (miRNA-7) on the sensitivity of cis-platin in esophageal carcinoma cell line TE-1, and the possible mechanism thereof. Methods Lipofectmin 2000 method was used to transient transfect with miRNA-7 mimic into esophageal cancer cell line TE-1, which was taken as transfection group, mimic negative control was taken as transfection conrtol group. The expressions of miRNA-7 and epidermal growth factor receptor (EGFR) mRNA were detected by RT-PCR in the above two groups and normal control group. The total EGFR and EGFR in cytoplasmic and nucleus were detected with Western blot assay in transfection group and transfection control group. CCK-8 was used to detect IC50 of cisplatin in transfection group and transfection control group. The expression of EGFR was observed with immunofluorescence confocal microscope in two groups. Results The miRNA-7 expression was signifi-cantly increased in transfection group than that of transfection conrtol group and control group. The expression of EGFR mRNA was significantly reduced in transfection group (P<0.001). The total EGFR was significantly decreased in transfec-tion group than that of transfection conrtol group. The level of nuclear EGFR was significantly increased ( P<0.01),and cyto-plasm EGFR expression was significantly decreased in transfection group than that of transfection control group ( P<0.05). CCK-8 results showed that after the over expression of miRNA-7 in TE-1, the IC50 of cisplatin (48 h) increased in transfec-tion group than that of control group (P<0.01). Immunofluorescence results showed that EGFG in nuclear was higher in transfection group than that of transfection control group but its expressions reduced in cell membrane and cytoplasm. Con-clusion The over-expressed miRNA-7 in esophageal cancer cells TE-1 can reduce cisplatin sensitivity by the increased EGFR in nuclear translocation.

Objective: To make it clear whether there exists m elatonin receptor in the thyroid of human embryo. Methods: Thyr oid was collected and sliced up to be stained with methods of immunohistochemis try and in situ hybridization. Results: The thyroid tissue w as p ositively dyed, melatonin receptor mt1 and MT2 were with both immunohistoche mistry and in situ hybridization while brown granules dep osited in the membrane, plasma and nuclear of the thyroid cell were with the imm unohitochemistry. Conclusion: There exists melatonin rece ptors in human embryo thyroid, either mt1 or MT2, and they exist in the memb rane, plasma and nuclear.

Background The experimental studies of venous thromboembolism (VTE) as an entity and the response of the pulmonary arterial endothelium after VTE are still rare.The objective of this study was to observe changes in the pulmonary arterial endothelium using a novel rat model of VTE.Methods Rats were allocated to the VTE (n=54) or control groups (n=9).The left femoral vein was blocked using a microvessel clip to form deep vein thrombosis (DVT).One,four or seven-day-old thrombi were injected into the right femoral vein to induce DVT-pulmonary thromboembolism (DVT-PTE).The rats were sacrificed 1,4 or 7 days later (Dn (1,4,7) Pn(1,4,7) subgroups (n=6)),and the lungs were examined using light and electron microscopy.Results On gross dissection,the rate of DVT formation was higher on day 1 (D1Pn:100％,18/18) than day 4 (D4Pn:83％,15/18; x2=5.900,P=0.015) or day 7 (D7Pn:44％,8/18; x2=13.846,P=0.000).On gross dissection,the positive emboli residue rate in the pulmonary arteries was lower in the D1Pn subgroup (39％,7/18) than the D4Pn (73％,11/15;x2=3.915,P=0.048) and D7Pn subgroups (100％,8/8; x2=8.474,P=0.004); however,light microscopy indicated the residual emboli rate was similar in all subgroups.Hyperplasia of the pulmonary arterial endothelium was observed 4 and 7 days after the injection of one-day-old or four-day-old thrombi.However,regions without pulmonary arterial endothelial cells and intra-elastic layers were observed one day after injection of seven-day-old thrombi.Conclusions This novel model closely simulates the clinical situations of thrombus formation and is ideal to study pulmonary endothelial cell activation.The outcome of emboli and pulmonary arterial endothelial alterations are related to the age and nature of the thrombi.

OBJECTIVETo investigate the effects of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells on collagen gel contraction in organotypic culture.

METHODSThe hair follicle organotypic culture was prepared with different concentrations of rat tail collagen, different number of dermal papilla cells and hair follicle epithelium cells in DMEM medium, after cultured for 10 days the diameter of collagen gel was measured.

RESULTThe concentration of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells significantly influenced on collagen gel contraction in organotypic culture (P<0.01). The contraction of collagen gel was negatively related to the concentration of rat tail collagen, while the concentration of dermal papilla cells and hair follicle epithelium cells was positively related to the contraction of collagen gel.

CONCLUSIONThe key factor influencing collagen gel contraction in organotypic culture is the concentration of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells.

Animals ; Cell Division ; Cells, Cultured ; Collagen ; physiology ; Gels ; Hair Follicle ; cytology ; Rats

OBJECTIVETo investigate the expression of bFGF, ET-1 and SCF in different passages of cultured dermal papilla cells (DPC), and their possible effect on biological behaviour of DPC.

METHODSThe expression of bFGF, ET-1 and SCF in different passages of cultured DPC was detected by immunocytochemistry and in situ hybridization.

RESULTThe expression of ET-1 and SCF in early passages of cultured DPC was stronger, but became negative in late passages (>6 passages). The stronger the expression of ET-1 and SCF in DPC, the higher ability of DPC to induce hair follicle regeneration.

CONCLUSIONThe expression strength of ET-1 and SCF is related to the ability of DPC inducing hair follicle regeneration.

Endothelin-1 ; analysis ; genetics ; Fibroblast Growth Factor 2 ; analysis ; genetics ; Hair Follicle ; chemistry ; cytology ; physiology ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Stem Cell Factor ; analysis ; genetics