AIM:To investigate the protective effect of nerve growth factor combined with Ginkgo biloba extract on retinal ischemia-reperfusion (RIR) injury in rabbits with experimental high intraocular pressure.METHODS:Establishment of rabbit glaucoma ischemia reperfusion model.Twenty-four New Zealand white rabbits were randomly divided into three groups:nerve growth factor group, Ginkgo biloba extract group and combination group.Respectively, in the continuous administration of 1, 7, 14d.We observed the morphological changes of the tissues of the retina.The levels of superoxide dismutase(SOD), nitric oxide(NO) and malondialdehyde(MDA) in retinal tissue were measured.RESULTS:Respectively, first, in the continuous administration of 1, 7, 14d, the contents of MDA and NO in Ginkgo biloba extract group and nerve growth group were higher than that in combination group (P<0.05).Secondly, the SOD content of Ginkgo biloba extract group and nerve growth group were lower than that of combination group at each time point (P<0.05).At each time point, the number of HE staining of retinal ganglion cells (RGCs) showed that the loss of RGCs in the combination group was significantly lower than that in the other groups, and the ganglion cell count showed that the Ginkgo biloba extract group and the neuronal growth group were lower (P<0.05).CONCLUSION:Nerve growth factor combined with Ginkgo biloba extract has better protective effect on retinal ischemia-reperfusion injury.The mechanism may be related to the decrease of free radicals and increase the activity of SOD in retinal tissue.

Adult ; Bronchoalveolar Lavage ; nursing ; Female ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; nursing ; therapy ; Retrospective Studies

Objective To study the effect of dialysis adcquacy,microinflammation and residual renal function on nutritional status of hemodialysis patients.Methods One hundred and fourteen patients were enrolled in this study.Kt/V,?_2-MG and serum iPTH were measured as markers of hemodialysis adequacy.Nutritional evaluation included MQSGA,Alb,Hb,TF,IGF-1,IGFBP-3 and anthropometrics such as HGS,BSF,TSF,MAC,MAMC and AMA.Serum IL-6,TNF-?and CRP were detected to assess microinflammation.Urinary volume of 24 hours was measured to investigate the residual renal function (RRF).Results (1)There were different correlations and regressive associations of Kt/V,iPTH and?_2-MG with HGS,MAMC,AMA,Alb,Hb,nPCR,IGF-1 and MQSGA respectively.(2) There were significant correlations and regressive associations of RRF to HGS,TSF,MAMC,Alb,nPCR and IGF-1 within the first year of hemodialysis.(3) There were different correlations and regression relationships of IL-6,TNF-?and CRP with HGS、MAMC、AMA、Alb、TSF、Hb、nPCR、IGF-1 respectively.(4) Multivariate analysis showed that Kt/V,iPTH,IL-6, TNF-?,?_2-MG and RRF were influencing factors,among them,Kt/V,iPTH,IL-6 and TNF-?were independent predictors of nutritional status.Conclusions Hemodialysis adequacy and micruinflammation may impact on nutritional status.Residual renal function may be involved in nutritional status in the first year of hemodialysis.Kt/V,iPTH,IL-6 and TNF-?are independent factors affecting nutritional status.

OBJECTIVETo explore the effects of garlic oil combined with resveratrol on the apoptosis and expression of Fas, bcl-2 and bax in human gastric cancer cell line MGC-803.

METHODSThe experiment included three groups which were the control group, the combined medicine group 1 (including 25 microg/ml oil garlic and 25 microg/ml resveratrol) and the combined medicine group 2 (including 50 microg/ml oil garlic and 50 microg/ml resveratrol). The apoptosis of cell was examined by DNA gel electrophoresis and flow cytometry for annexin v; the expression of Fas was determined by flow cytometry at the 24th hour after the treatment; the mRNA expression of bcl-2 and Bax gene were measured by RT-PCR method at the 24th ang 48th after treatment, respectively.

RESULTSThe garlic oil combined with resveratrol induced cell apoptosis markedly at the 24th after the treatment The protein expression of Fas in the combined medicine groups was 10.59% and 14.16% respectively. As compared with the control group (5.27%), the statistical significance was obvious. The mRNA level of Bax was elevated significantly, however the mRNA expression of bcl-2 was decreased at the 24th and 48th after the treatment.

CONCLUSIONSThe garlic oil combined with the resveratrol might obviously induce the apoptosis of gastric cancer cell line MGC-803 which be involved in increasing the expression of Fas protein and bax gene and decreasing the expression of bcl-2 gene at the same time.

Apoptosis ; drug effects ; genetics ; Cell Line, Tumor ; Garlic ; Gene Expression Regulation, Neoplastic ; drug effects ; Genes, bcl-2 ; Humans ; Plant Oils ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; Stilbenes ; pharmacology ; Stomach Neoplasms ; genetics ; bcl-2-Associated X Protein ; genetics ; fas Receptor ; genetics

AIM: To observe the effects of the human acidic fibroblast growth factor mutant (mhaFGF), lacking 27 amino acids at N-terminal, on the proliferation and signal transduction of the hepatocarcinoma cells. METHODS: The hepatocarcinoma cells were treated with human acidic fibroblast growth factor (haFGF) and mhaFGF, respectively. The expression levels of the signal proteins, Grb2 and Erk1/2, in the hepatocarcinoma cells were detected by semi-quantitative Western-blotting after treated for 15 min. The mitogenic activity of both haFGF and mhaFGF was detected by MTT method and the cell cycle was analysed by flow cytometer (FCM) after treated for 48 h. RESULTS: The mitogenic activity and the ratio of G 1 and S phase cells in mhaFGF-treated cells were markedly lower than that of the haFGF, and close to that of the control group. The expression level of both Grb2 and Erk1/2 in the mhaFGF-treated cells were lower than those in the haFGF- treated cells. CONCLUSION: The decrease in the mitogenic activity of mhaFGF is probably associated to its down-regulating the expression of the signal molecular, MAPK-ERK1/2 and Grb2.

AIM:The present study was designed to establish H2O2-induced NRK52E cell apoptotic model and to investigate the effects and the mechanism of nmhaFGF on the NRK52E cell apoptosis induced by H2O2. METHODS: In vitro experiment, a apoptotic model of normal rat kidney epithelium (NRK52E) was made by MTT method, Hoechst33342 dyeing and flow cytosorting (FCR). NRK52E cells were cultured with different concentrations of nmhaFGF and haFGF for 24 h before H2O2 was added. The apoptotic rate was detected with FCR method. RESULTS: H2O2 at concentration of 0.4 mmol/L, the optimal condition to establish the apoptotic model, was used to treat NRK52E cells for 18h. Different doses of nmhaFGF (0.01, 0.03, 0.10, 0.30, 1.00 mg/L) reduced the apoptotic rates with the dose rising. However, the decreasing tends of apoptotic rates with dose increasing for haFGF was not so obvious. CONCLUSION: nmhaFGF protects the NRK52E cells against apoptosis. The mechanism might be connected with its non-mitogenic property.

AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.

AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.