Objective To explore the effects of interleukin- 1?(IL-1?)on the biological activity of human periodontal ligament(PDL) cells in vitro. Methods Human PDL cells were cultured in DMEM medium containing IL-1?(0.1,0.5,1,5 and 10ng/ml) for 1,2,3,4 and 5 days, respectively. The proliferation of PDL cells was measured by MTT assay at the first, second, third, fourth and fifth days after IL-1? treatment, respectively. Fibronectin level in the medium was determined by ELISA at the fourth days after 10ng/ml IL-1? treatment, and alkaline phosphatase(ALP) activity was measured by enzyme kinetic method. Results IL-1? inhibited the growth of human PDL cells in a dose-dependent manner, and its lowest effective concentration was 1ng/ml(P

PurposeSmall breast mass (diameter≤1 cm) is prone to misdiagnosis in clinic. This paper aims to evaluate a combined application of breast imaging reporting and data system (BI-RADS) and ultrasound elastography (UE) on small breast mass (diameter≤1 cm).Materials and MethodsA retrospective analysis was carried out on 231 patients with a total of 258 small masses (the maximal diameter ≤1 cm). Ultrasound BI-RADS was used for classiifcation while UE was used to adjust the results. The results were further compared with those of postoperative pathology. The curve of ROC was employed to evaluate the combined use on small breast mass.ResultsAmong the 258 small masses, 178 (69.0%) were benign masses and the rest 80 (31.0%) were malignant. The small masses which were evaluated as BI-RADS grade 3, 4 and 5 before the operation had positive prediction value for malignant masses of 10.3% (17/165), 60.5% (46/76) and 100.0% (17/17), respectively. After adjustment with UE, the values changed to 5.3%(9/169), 75.0% (54/72) and 100.0% (17/17), respectively. After adjustment with the combination method, the area under ROC curve in BI-RADS classification was 0.904, which was signiifcantly higher than that (0.827) before the adjustment (Z=2.83,P<0.05). ConclusionFor small breast mass (diameter≤1 cm), mass of BI-RADS grade 3 has higher positive prediction value. But after adjustment with UE, the positive prediction value of mass of BI-RADS grade 3 tends to be lower, whilst that of mass of BI-RADS grade 4 increases, thus promoting the efficiency of ultrasound BI-RADS classification for small breast mass and contributing to the identiifcation of benign and malignant small breast masses.

Objective To investigate whether the inhibition of caveolin-1 (Cav-1) phosphorylation will regulate effectively nuclear factor-erythroid 2-related factor (Nrf2) signal pathway and downstream effector molecules and protest against ventilation induced lung injury (VILI) in an animal model in vivo. Methods Ninety male Sprague-Dawley (SD) rats were randomly divided into nine groups (each n = 10): sham group in which rats did not receive ventilation but received tracheotomy; lung protective ventilation (PV) for 1 hour or 2 hours group; mechanical ventilation (MV) at high volume tidal (VT, 40 mL/kg) for 1 hour or 2 hours group; protein tyrosine kinase inhibitor PP2 or rosiglitazone (Rsg) pretreatment + high VT ventilation for 1 hour or 2 hours groups. The two pretreatment groups were given intraperitoneal injection PP2 15 mg/kg or intragastric administration of Rsg 5 mg/kg 1 hour before ventilation respectively. The rats were sacrificed after model reproduction, and bronchoalveolar lavage fluid (BALF) was collected. Pulmonary vascular permeability was measured by Evans blue (EB). The levels of tumor necrosis factor-α (TNF-α), activator protein-1 (AP-1), nuclear factor-κB (NF-κB), and interleukin-8 (IL-8) in BALF were determined by enzyme linked immunosorbent assay (ELISA). Then the lung tissues were collected, the lung wet/dry ratio (W/D) was calculated, the changes in pathology was observed with light microscope, and myeloperoxidase (MPO) activity was determined by colorimetric analysis. Nrf2 mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of Cav-1 tyrosine residues 14 phosphorylation (pCav-1-Y14), Cav-1, peroxisome proliferators-activated receptor γ (PPARγ) and claudin-5 as well as Nrf2 in cytoplasm and nucleus were determined by Western Blot. The positive expressions of PPARγ and claudin-5 in lung tissues were assayed with immunohistochemistry staining. Results There were no obvious pathological changes in the lung tissue in sham group and PV groups, and there were no significant differences in all the parameters between the two groups either. However, the injury in lung tissue was severe in the high VT groups in which W/D ratio, EB contents, MPO activity, and TNF-α, AP-1, IL-8, NF-κB levels in BALF as well as the protein expressions of Cav-1 and pCav-1-Y14 were significantly higher than those of sham group and PV groups, and the protein expressions of PPARγ and claudin-5 were significant lower than those of sham group and PV groups with a dose-dependent manner; but Nrf2 expressions in cytoplasm and nucleus did not show a statistical increase. After pretreatment of PP2 or Rsg, W/D ratio, MPO activity, EB contents, TNF-α, AP-1, IL-8, and NF-κB in BALF were significantly decreased as compared with those of high VT group, and RT-PCR showed significant up-regulation of Nrf2 mRNA in lung tissues too. Moreover, there was a statistically significant increase in expressed Nrf2 proteins in nucleus in PP2 or Rsg groups as compared with those of high VT groups [Nrf2 in nucleus (gray value): 0.61±0.06, 0.56±0.06 vs. 0.31±0.02 at 1 hour, 0.38±0.06, 0.43±0.07 vs. 0.22±0.03 at 2 hours; all P < 0.05], but no significant difference was found in the expression of Nrf2 protein in the cytoplasm among all groups. The protein expressions of pCav-1-Y14 in PP2 pretreatment groups were significantly lower than those of high VT groups (gray value: 0.89±0.04 vs. 1.48±0.02 at 1 hour, 0.86±0.02 vs. 1.31±0.01 at 2 hours; both P < 0.05); but expressed PPARγ proteins and expressed claudin-5 proteins in PP2 or Rsg pretreatment groups were significantly higher than those of high VT groups [PPARγ (gray value): 0.34±0.07, 0.42±0.13 vs. 0.17±0.07 at 1 hour, 0.38±0.09, 0.33±0.07 vs. 0.16±0.03 at 2 hours; claudin-5 (gray value): 0.33±0.05, 0.38±0.07 vs. 0.14±0.03 at 1 hour; 0.30±0.06, 0.31±0.04 vs. 0.17±0.04 at 2 hours; all P < 0.05]. Conclusions The inhibition of Cav-1-Y14 phosphorylation can increase the expression of Nrf2 in the nucleus, then result in an increase in the protein expressions of PPARγ and claudin-5 of its effector molecules. This effect can reduce the inflammation and capillary permeability of lung tissue in the model of VILI.

ObjectiveTo determine whether the inhibition of caveolin-1 tyrosine residues 14 (Cav-1-Y14) phosphorylation with protein tyrosine kinase inhibitors (PP2) will upregulate heme oxygenase-1 (HO-1) activity to protect against ventilation induced lung injury in vivo of an animal model.Methods Fifty-four male Sprague-Dawley (SD) rats were randomly divided into nine groups (eachn = 6). Group A served as normal control group, in which rats did not receive ventilation but tracheotomy. Groups B1 and B2 received lung protective ventilation respectively for 1 hour or 2 hours. Groups C1 and C2 received high tidal volume (40 mL/kg) ventilation for 1 hour or 2 hours, respectively. The group D1 or D2 also received high tidal volume ventilation for 1 hour or 2 hour respectively, but they were given PP2 1 hour before high tidal volume ventilation. The groups E1 and E2 also received high tidal volume ventilation respectively for 1 hour or 2 hours, but tyrosine kinase inhibitor PP2 and HO-1 inhibitor zinc protoporphyrinⅨ(ZnPPⅨ) were given to animals 18 hours before high tidal volume ventilation. All the animals were sacrificed after ventilation, and the specimens of lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Then the changes in pathology of lung tissue was observed, and diffuse alveolar damage scores (DAD) were calculated, myeloperoxidase (MPO) activity was measured by colorimetric analysis, lung wet/dry ratio (W/D) was estimated. The expressions of phosphorylated caveolin-1 (P-Cav-1-Y14), caveolin-1 (Cav-1) and HO-1 were determined by Western Blot. The expressions of high mobility group B1 (HMGB1) and advanced glycation end product receptor (RAGE) in lung tissues were assayed with immunohistochemistry staining. The levels of tumor necrosis factor-α(TNF-α) in BALF were measured by enzyme linked immunosorbent assay (ELISA).Results There was no significant difference in all the parameters between group A and groups B. Compared with group B1, DAD score, W/D ratio, the activity of MPO and the concentration of TNF-α in BALF in group C1 were significantly increased [DAD score:7.97±0.59 vs. 0.55±0.13, W/D ratio: 5.70±1.61 vs. 5.04±0.63, MPO (U/g): 1.82±0.14 vs. 0.77±0.26, TNF-α(ng/L): 370.10±29.61 vs. 54.38±8.18, allP< 0.05], and the injury in ventilation 2 hours group was more serious than that in ventilation 1 hour group. Compared with groups C, all the parameters in groups D were significantly decreased. The parameters in groups E were significantly higher than those in groups A, B, and D, but no significant difference was found as compared with groups C. Compared with groups B, the protein expressions of Cav-1 and P-Cav-1-Y14 (gray value) in groups C were significantly increased (1 hour: 1.49±0.02 vs. 1.26±0.13, 1.34±0.02 vs. 0.87±0.04;2 hours: 1.58±0.02 vs. 1.27±0.27, 1.31±0.01 vs. 0.95±0.02, allP< 0.05), and the expression of HO-1 protein (gray value) was significantly decreased (1 hour: 0.59±0.02 vs. 1.10±0.01, 2 hours: 0.49±0.01 vs. 1.20±0.02, both P< 0.05). No significant difference in Cav-1 protein expression between groups D as well as groups E and groups C. The protein expression of P-Cav-1-Y14 in groups D and E was significantly lower than that in groups C. The protein expression of HO-1 in groups D was significantly higher than that in groups C, but the phenomenon was not found in groups E as compared with groups C. Compared with group A, the positive expression of HMGB1 and RAGE in lung tissue in groups C and E was significantly increased, but no significant difference was found between groups B as well as groups D and group A.Conclusion Cav-1-Y14 phosphorylation is the key factor for ventilator induced lung injury, which can not only lead to a decrease in vascular barrier function, but also inhibit the activity of HO-1 enzyme, thus further aggravates inflammatory injury of the lung as induced by mechanical ventilation.

Objective To express EV71 VP1 in prokaryotic expression system,initially evaluate the ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody.Methods Construct the prokaryotic expression plasmid pET30a (+)-VP1.Induced expression in Transetta (DE3) with IPTG and identified by Western blot.After purified with HisBind Protein Purification Kit,its ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody were analyzed.Results Plasmid PET-30a(+)-VPI was constructed successfully and the objective protein was expressed effectively.The ELISA titer of the polyclonal antibody was 1:3200 while neutralizing titer was 1:16 and the recombination protein lost the ability of blocking EV71 infection.Conclusion The recombination protein can stimulate mice to produce antibodies and the polyclonal antibody shew neutralizing activity but the recombination protein lost the binding activity to receptors probably due to the wrong advanced structure.

ObjectiveTo explore the effect of the calcium phosphate cement (CPC) /calcium polyphosphate fiber (CPPF) composites mixed with different proportion of minimal morselized bone on repair of bone defect in vivo. MethodsA total of 36 New Zealand white rabbits were completely randomly designed into A, B, C, D groups and their bilateral radial bone defect model was prepared. The minimal morselized bone (300-500 μm in diameter) was made from the iliac of those rats. The CPPF and CPC were evenly mixed into CPC/CPPF composites which were divided into four groups in accordance with the CPPF weight O, 10％, 30％ and 50％ in CPC/CPPF composite. The CPC/CPPF composites of the four groups was mixed with the minimal morselized bone with ratio of 6:4 and then the mixture was implanted the bone defect of the rabbits in four groups. The gross, X-ray and histological observations were done at four and eight weeks. The biomechanical test was performed at eight weeks. Results When CPPF occupies 30％ of the CPC/CPPF composite, the maximum compressive load and bending loads were better than those in the other groups ( P ＜ 0.05 ), when the histological observation showed the most tight link between the artificial composite and the bone interface and the closest similarity between material degradation rate and the ossification rate, with the best osteogenesis and the optimal ratio.ConclusionThe repair of bone defect can attain the optimal outcome through adding a certain ratio of minimal morselized bone into the CPC/CPPF to adjust the degradation rate of composites.

Objective To explore the clinical significance of epithelial cellular adhesion molecule (Ep-CAM)expression in esophageal carcinoma.Methods Ep-CAM expression of in 60 cases of esophageal carcinoma、35 cases of atypical hyperplasia and 35 cases of normal esophageal mu-cosa was respectively examined by immunohistochemistry.Results Ep-CAM expression were ob-served with positive in 28.75% of atypical hyperplasia and 90.00% of esophageal carcinoma,but with no positive expression in normal esophageal mucosa.The Ep-CAM expression with positive were significantly differently between degrees of tumor differentiation(P <0.01),lymphatic metas-tasis and 3-year survival(P <0.05).Conclusion The increase of Ep-CAM expression,as a specific index,may has the potential for esophageal cancer diagnosis and prognosis.

Objective To explore the clinical significance of epithelial cellular adhesion molecule (Ep-CAM)expression in esophageal carcinoma.Methods Ep-CAM expression of in 60 cases of esophageal carcinoma、35 cases of atypical hyperplasia and 35 cases of normal esophageal mu-cosa was respectively examined by immunohistochemistry.Results Ep-CAM expression were ob-served with positive in 28.75% of atypical hyperplasia and 90.00% of esophageal carcinoma,but with no positive expression in normal esophageal mucosa.The Ep-CAM expression with positive were significantly differently between degrees of tumor differentiation(P <0.01),lymphatic metas-tasis and 3-year survival(P <0.05).Conclusion The increase of Ep-CAM expression,as a specific index,may has the potential for esophageal cancer diagnosis and prognosis.

OBJECTIVETo investigate the influences of temperature, lightness, storage method, storage time, and gibberellin on seed germination of Gentiana rigescens.

METHODThe germination rates of G. rigescens in different treatments were observed.

RESULT AND CONCLUSIONThe most suitable temperature for the seed germination was 25 degrees C, at which the germination rate was 76.33%. The effect of lightness on the seeds was significantly; the germination rate of the seed was very low. Under the natural condition, the best storage method was dry storage (within 6 months), which could promote the after-ripening of the seed. 100-1 000 mg x L(-1) gibberellic acid could significantly reduce the seed germination time, and 500 mg x L(-1) gibberellic acid increased the germination rate of the seed to 95.00%.

Gentiana ; drug effects ; growth & development ; Germination ; drug effects ; Gibberellins ; pharmacology ; Plant Growth Regulators ; pharmacology ; Seeds ; drug effects ; growth & development ; Sunlight ; Temperature

Objective To investigate the therapeutic mechanism of Gandou Bushen Decoction(GDBSD)for improving reproductive disorders in male mouse models of Wilson disease(WD).Methods Sixty male homozygous TX mice were randomized equally into 4 groups and treated with daily gavage of saline(WD model group),penicillamine(0.09 g/kg),or GDBSD(0.2 mL/10 g),or with intraperitoneal injection of U0126(20 mg/kg)in addition to GDBSD gavage,with 15 male DL mice as control.After 4 weeks of treatment,copper content in testicular tissue of the mice was detected,and histopathology of the testes and epididymis was examined using HE staining and electron microscopy.TUNEL staining was used to identify apoptotic cells in the testes.The protein expressions of Bcl-2,Cytc,caspase-3,ERK,and p-ERK in the testicular tissue were evaluated with Western blotting,and BrdU-positive cells were detected with immunohistochemical labeling.Sperm density,viability,malformation rate and fertility levels of male mice were studied.Results Treatment with penicillamine and GDBSD obviously improved pathological changes of the testis,increased sperm density and motility,lowered sperm abnormality rate,fertility levels and increased testicular JOHNSEN score of TK mice,but the therapeutic effect of GDBSD was blocked by U0126.GDBSD treatment significantly lowered Cytc and caspase-3 expressions and increased Bcl-2 expression in the testicular tissue of TX mice(P<0.05),while U0126 treatment significantly lowered testicular Bcl-2 expression level.No significant differences were found in total protein expression levels of ERK1/2 among the 5 groups,but p-ERK protein expression was significantly reduced in WD and U0126 groups and increased in penicillamine and GDBSD groups.Conclusion GDBSD can improve spermatogenesis and enhance fertility of male TX mice with WD possibly by activating the ERK signaling pathway to enhance proliferation and reduce apoptosis of the spermatogenic cells.