AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

OBJECTIVETo investigate the protective effect of Shenfu Injection against ischemia-reperfusion (I/R) injury due to pancreas transplantation in rats, and explore its possible mechanism.

METHODSSix normal SD rats with sham operation were taken as the normal control group, 24 steptozozin-induced diabetic SD rats were randomly divided into 4 groups, with 6 in each group. Except I/R group, the rats in the other groups were intravenous injected with Shenfu Injection (SF,10 mg/kg), Hongshen Injection (HS, 9 mg/kg) and Fuzi Injection (FZ 1 mg/kg) respectively at the day and 30 minutes before pancreas transplantation performed in the SF group, HS group and FZ group, respectively. At the same time, rats in the normal control group and in the I/R group were intravenously injected the same volume of normal saline. The blood glucose was detected before and after reperfusion, and 2 hours later after reperfusion, the contents of serum nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), the concentrations of malondialdehyde (MDA) , superoxide dismutase (SOD) , and myeloperoxidase (MPO) in the transplanted pancreas tissues were detected. The cell apoptosis of the transplanted pancreas tissue was determined by TUNEL, and the bcl-2 and Bax protein expression was determined by Western blot.

RESULTSAfter reperfusion, the levels of blood glucose and TNF-alpha decreased and the concentration of NO increased in the SF group, HS group and FZ group, compared with those in the I/R group. The activity of SOD, bcl-2 expression and the ratio of bcl-2 and Bax were higher, while the content of MDA, the activity of MPO, apoptotic indexes, and Bax expression were lower in the SF group, HS group and FZ group than those in the I/R group.

CONCLUSIONShenfu Injection can protect L/R injury due to pancreas transplantation in rats, the possible mechanism may be related to promoting activity of SOD, increasing synthesis of endogenous NO, decreasing the excretion of TNF-alpha, alleviating conglutination and aggregation of polymorphonuclear neutrophils (PMNs) in pancreas, as well as up-regulating Bcl-2 gene expression and down-regulating the Bax gene expression.

Animals ; Cell Aggregation ; drug effects ; Diabetes Mellitus, Experimental ; enzymology ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Injections ; Malondialdehyde ; metabolism ; Nitric Oxide ; blood ; Pancreas ; drug effects ; metabolism ; Pancreas Transplantation ; adverse effects ; Protective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; prevention & control ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Animals ; Electromagnetic Fields ; Embryo, Mammalian ; metabolism ; Female ; Mice ; Pregnancy ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To explore the protective effcets of Schisandra chinensis extract (SCE) in paraquat (PQ)-induced pulmonary fibrosis in mice ,and its intrinsic molecular mechanisms thereof. Methods A total of 108 mice were randomly allocated into six groups (n=18):control group, model group, low concentration of SCE group (200 mg/kg), medium concentration of SCE group (400 mg/kg), high concentration of SCE group (800 mg/kg) and vitamin C group (100 mg/kg). Except control group, mice were given by intragastric administration with PQ (100 mg/kg) and administered with SCE and Vitamin C once per 24 h after PQ modeling. Mice were sacrificed at 7, 14 and 21 d after modeling. Six mice were executed at different time points. The degree of lung tissue inflammation and fibrosis were observed by HE staining and Masson staining. The mRNA and protein expression levels of transforming growth (TGF)-β1, interleukin (IL)-6 and IL-17 in lung tissue were determined by RT-PCR and ELISA respectively. Results (1) Compared with control group, the lung tissue of model group showed a large number of inflammatory cell infiltration, space congestion, and its inflammation scores increased at 7 and 14 days after modeling (P<0.05). At the same time, compared with model group and vitamin C group, inflammation scores were significantly decreased in medium concentration of SCE group and high concentration of SCE group (P<0.05). (2) Compared with control group, collagen fibers and the degree of fibrosis were significantly increased in model group ,while pulmonary fibrosis were decreased in medium concentration of SCE group and high concentration of SCE group at 14 and 21 days after modeling (P<0.05). (3) With the extension of modeling time, both mRNA and protein expressions of TGF-β1 were obviously elevated, IL-6 decreased and IL-17 reduced after the first increase in PQ group. Compared with PQ group, levels of three cytokines mRNA and protein expression in medium concentration of SCE group and high concentration of SCE group changed as follows:IL-6 level was markedly decreased at 7 and 14 days after modeling;TGF-β1 level was markedly increased at 14 and 21 days after modeling. However, IL-17 level was markedly decrease at three time points(P<0.05). Conclusion SCE can relieve PQ-induced lung inflammation and fibrosis by suppressing TGF-β1, IL-6, and IL-17 expressions.

OBJECTIVETo investigate the effect of celecoxib on adhesion, invasion, migration and matrix metalloproteinase-2(MMP-2) expression of tongue squamous cell carcinoma cell line Tca8113 cells.

METHODSFollowing incubation with celecoxib, the Tca8113 cells were detected for cell adhesion and migration using cell adhesion assay and Boyden chamber invasion assay. The expression of cyclooxygenase-2 (Cox-2) protein in Tca8113 cells was tected with SP immunohistochemistry staining. The MMP-2 level in supernatant was detected with enzyme-linked immunosorbent assay. RESULTS; The adhesion and Boyden chamber invasion assays showed that, after treatment celecoxib, the ability of adhesion and migration of Tca8113 cells was significantly inhibited. Celecoxib could decrease the expression of Cox-2 protein in Tca8113 cell and decrease the MMP-2 level in supernatant.

CONCLUSIONCox-2 inhibitor celecoxib can significantly inhibit the adhesion and migration of Tca8113 cells. The inhibitory effect on hesion and migration may be correlative with its effect on decrese of Cox-2 protein expression and secretion MMP-2 in Tca8113 cells.

Carcinoma, Squamous Cell ; Celecoxib ; Cell Adhesion ; Cell Line, Tumor ; Cyclooxygenase 2 Inhibitors ; Humans ; Matrix Metalloproteinase 2 ; Pyrazoles ; Sulfonamides ; Tongue Neoplasms

OBJECTIVETo investigate the effect of the extremely low frequency pulsed electromagnetic field (PEMF) on the proliferation and differentiation of osteoblast-like cells.

METHODSThe MC3T3-E1 cell and the primary osteoblast cell derived from 2-day-old Sprague Dawley (SD) rat calvaria were exposed to PEMF with a magnetic flux density of 1.55 mT at 48 Hz for 24 or 48 h. MTS was applied to analyze cell proliferation and flow cytometry to detect cell cycle. The intracellular alkaline phosphatase (ALP) activity was measured by colorimetry.

RESULTSPEMF of 1.55 mT at 48 Hz decreased significantly the cell percentage of S or G(2)M phase (P < 0.05), but did not affect cell number of MC3T3-E1 cells. Although the number of the primary osteoblast cells did not alter by MTS assay after exposure to PEMF for 24 h continuously, the cell percentage of G(2)M phase increased significantly (P < 0.01). When the culture time extended to 48 h, the cell number increased greatly (P < 0.01) and the cell percentage of G(2)M phase decreased significantly despite of the exposure type (P < 0.01). After the primary osteoblast cells were exposed to PEMF for 24 h continuously, the ALP activity decreased significantly (P < 0.05), whereas it increased significantly after exposure to PEMF for 48 h continuously (P < 0.05).

CONCLUSIONPEMF of 1.55 mT at 48 Hz does not affect proliferation and differentiation of MC3T3-E1 cell, but it promotes proliferation of primary osteoblast cell, inhibits differentiation at proliferation stage and promotes differentiation at differentiation stage of primary osteoblast cell.

Animals ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Dose-Response Relationship, Radiation ; Electromagnetic Fields ; adverse effects ; Mice ; Osteoblasts ; cytology ; metabolism ; radiation effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of extremely low frequency magnetic fields on intracellular calcium concentration ([Ca(2+)]i).

METHODSFura-2 loaded HepG2 cells were exposed to 1.55 mT (average value), 16 Hz pulsed magnetic fields for 60 min and to 300 mT, 2 Hz rotating magnetic fields for 5 min, and then [Ca(2+)]i was measured by fluorescence spectrophotometer. [Ca(2+)]i of HepG2 cells was also measured when they were exposed to 0.9 mT [root mean square (rms)], 16 Hz sinusoidal magnetic fields in real time.

RESULTSThe R values (F(340) nm/F(380) nm) of the control and the exposed group were 2.4519 +/- 0.2378 and 2.5266 +/- 0.2915 respectively after HepG2 cells were exposed to 1.55 mT, 16 Hz magnetic fields, 1.365 0 +/- 0.0626 and 1.3602 +/- 0.0771 respectively to 300 mT, 2 Hz rotating magnetic fields. The ratios of the trendline slope [r((501 - 1,000)) / r((0 - 500))] from the data of R values were 1.1213 +/- 0.4559 and 1.0727 +/- 0.1971 respectively (P > 0.05), and the ratios of the intercept [b((501 - 1,000)) / b((0 - 500))] from the trendline were 0.9912 +/- 0.0098 and 0.9979 +/- 0.0060 (P > 0.05) when HepG2 cells were exposed to the 0.9 mT, 16 Hz sinusoidal magnetic fields.

CONCLUSIONThe effect of extremely low frequency magnetic fields on [Ca(2+)]i of HepG2 cells under the experimental condition has not been found.

Calcium ; metabolism ; Cell Line, Tumor ; drug effects ; metabolism ; radiation effects ; Chelating Agents ; pharmacology ; Egtazic Acid ; pharmacology ; Electromagnetic Fields ; Humans ; Ion Transport ; drug effects ; radiation effects ; Octoxynol ; pharmacology ; Spectrometry, Fluorescence ; Time Factors

OBJECTIVETo study the effects of low frequency pulsed magnetic field on the proliferation and differentiation of HepG2 cells.

METHODS3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetry method and ELISA assay of alpha-fetoprotein (AFP) were used to determine the cell proliferation and differentiation after the cells were exposed to pulsed magnetic fields with different frequency but the same field intensity.

RESULTSThere were no significant differences in cell proliferation between sham and treated groups exposed to the field of 80 Hz, 1.55 mT for 1, 4, 8, 12, 24 h (P > 0.05). There were also no significant differences in cell proliferation and AFP secretion between sham and treated groups exposed to 16 Hz, 1.55 mT pulsed magnetic fields for 1, 4, 8, 24 h (P > 0.05).

CONCLUSIONThere were no "window effects" found in HepG2 cells proliferation or AFP secretion at 16 Hz and 80 Hz pulsed magnetic fields.

Cell Differentiation ; radiation effects ; Cell Division ; radiation effects ; Cell Line, Tumor ; cytology ; metabolism ; radiation effects ; Electromagnetic Fields ; Humans ; alpha-Fetoproteins ; analysis

Objective To investigate the imaging manifestations of lung neoplasms in pre- and post- treatment with CT-guided Argon-Helium cryoablation.Methods All the lung neoplasms in 96 patients have been treated with CT-guided percutaneous Argon-Helium targeted cryoablation.All patients have pre- and post-treatment CT scanning in measurement of lesion size and CT value.The CT scanning has been rerpeated afterl,3,6,12 months of treatment.Results Among total 96 cases,there are 82 cases of lung cancer and 14 cases of metastasis with 110 lesions(89 cases with single lesion,7 cases with multiple lesions).The Ar-He cryoablation has been given 103 times in total.The size of each lesion ranged from 1.2 cm to 15.0 cm in diameter with mean value of(4.0?2.5)cm,including 12 lesions less than 2 cm,51 lesions between 2— 4 cm,24 lesions between 4—6 cm,23 lesions over 6 cm.There are 25 patients whose lesions covered by iceball with 1 cm of overloaping it's margin.There are 63 lesions with diameter less than 4 cm gained 100% ablation rate,24 lesions with 4—6 cm diameter gained 95.8% ablation rate,and 23 lesions with over 6 cm diameter gained 69.6% ablation rate.The post-treatment CT show a progressively enlarged round,low density refrigerant area which clearly demarked with non- refrigerant area.The center of each refrigerant area has negative CT value,the mean decreased CT value of lesion instantly after the treatment are about 30— 50 HU with P

AIMTo observe the effect of FAK-related non-Kinase (FRNK) plasmid on hepatic stellate cell (HSC) proliferation stimulated by fibronectin (FN).

METHODSFRNK plasmid was transfected into HSC with transient liposomal transfection. The proteins of FRNK, FAK and p-FAK(Tyr397) were assayed by Western blotting analysis. The proliferation of HSC was evaluated by improved MTT assay, and cell cycle pattern was determined by flow cytometry (FCM).

RESULTS(1) The expression of FRNK protein increased after FRNK transfected HSC, and it was at 48 h that the expression of FRNK protein was the highest (P < 0.01). The protein level of FAK was no significant difference between before FRNK plasmid transfection and after transfection (P > 0.05). The expression of p-FAK(Tyr397) protein was down-regulated after FRNK had been transfected in HSC, (P < 0.01). (2) The HSC proliferation inhibition rates at 12 h, 24 h and 48 h after FRNK transfection were 20.07%, 26.16%, 29.77%, respectively (P < 0.01). (3) Compared with the non-FRNK plasmid group, the FRNK-transfected HSCs almost arrested in G0/G1 phase (71.4 +/- 2.81 vs 48.9 +/- 1.66, P < 0.01).

CONCLUSIONAfter FRNK were transfected successfully in HSCs using lipofectamine, the phosphorylation of FAK was inhibited. The HSC proliferation was restrained in a time-dependent manner and the HSC was arrested in G0/G1 phase.

Cell Line ; Cell Proliferation ; Fibronectins ; Hepatic Stellate Cells ; cytology ; Humans ; Phosphorylation ; Plasmids ; genetics ; Protein-Tyrosine Kinases ; genetics ; Transfection