Objective To develop a highly sensitive and accurate time-resolved immunofluorometric assay (TR-IFMA) for measurement of Dickkopf-1(DKK-1),as a novel serologic biomarker for lung cancer. Methods The study constructed a two-manoclonal-antibody "sandwich"-type assay and the sensitivity, within run CV and between run CV and accuracy were evaluated. Serum DKK-1 concentrations were measured by TR-IFMA in 120 healthy controls, in 72 benign lung disease patients, and in 212 lung cancer patients before surgery. The association between serum DKK-1 levels and clinicopathological features were evaluated. Results A standard curve for DKK-1 TR-IFMA had been developed with good sensitivity (0.08 μg/L). Both within run CV and between run CV were less than 6.5%. Accuracy studies, parallelism and precision data were determined and all found to be satisfactory. The validity of DKK-1 assay was confirmed by the good correlation between the results obtained by TR-IFMA and commercial ELISA (r=0.972, P=0.01). The serum levels of DKK-1 were higher in lung cancer patients 31.93(79.47-18.03) μg/L than in benign lung diseases 15.25(18.41-11.49) μg/L and in healthy controls 13. 90( 16. 91-11.02) μg/L DKK-1 levels were significant associated with the presence of distant metastases, as well as lymph node metastases and TNM stage, but not with patients' age, gender and tumor histology. At the cutoff of 22.63 μg/L, the diagnostic sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the TR-IFMA for lung carcinoma were 68.4%, 92.2%, 82.1% ,90.6% and 72.5%. Diagnostic sensitivity and accuracy were higher for small cell carcinoma than for non-small cell carcinoma (70.7% vs 69.5% and 85.6% vs 80.7%, respectively). Conclusions A highly sensitive and reliable TR-IFMA for DKK-1 has been developed. The determination of serum DKK-1 levels may be useful for diagnosis and tumor staging of lung cancer.

Chitinases genes from Metarhizium anisopliae which is an important entomopathogenic fungus were considered one of the key factors to invade their hosts. One Metarhizium anisopliae HN1 strain was isolated and screened. A chitinase gene was amplified by RT-PCR from Metarhizium anisopliae HN1, The whole length of this gene was 1275bp,and the nucleotide sequence of the gene was 96% similarity to that of the M. anisopliae E6 accessed in GenBank ( AF02749). The gene has been registered in GenBank and its accession number is DQ011865. The gene was subcloned into prokaryon expression vector pET-22b( + ), transformed this recombinant expression plasmid into E. coli strain BL 21 and effective expressed. The SDS-PAGE analysis indicated that the recombinant protein was 42kDa which is same to the reported article. The expression level of recombinant protein was about 63. 3% of whole expressed proteins , And when recombinant E. coli were crushed by freeze and supersonic wave , the activity assay indicates that the chitinase expressed in bacteria possesses biological activity.

OBJECTIVETo explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS).

METHODSWe randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR.

RESULTSThe concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05).

CONCLUSIONOrnidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

Animals ; Antioxidants ; Asthenozoospermia ; chemically induced ; drug therapy ; metabolism ; Carnitine ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Epididymis ; metabolism ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oligospermia ; chemically induced ; drug therapy ; metabolism ; Ornidazole ; Random Allocation ; Rats ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; alpha-Glucosidases ; metabolism

Objective To assess the left ventricular function of patients with acute myocardial infarction (AMI) treated by percutaneous coronary intervention (PCI) by speckle tracking imaging (STI).Methods 75 AMI patients who had AMI for the first time and have been treated by primary PCI were enrolled.Dynamic images were acquired before PCI,at 6 months after PCI and analyzed by STI.Dynamic images were analyzed for longitudinal peak systolic strain (LPSS),radial peak systolic strain (RPSS) and circumferential peak systolic strain (CPSS) values by STI.According to the comparison of left ventricular ejection fraction (LVEF) before PCI and 6 months after PCI,patients were divided into left ventricular function improved group (△LVEF≥5％) and not-improved group.Results Compared to non-improved group,LPSS (P ＜0.001),RPSS (P ＜0.05,P ＜0.001) and CPSS (P ＜0.001) of improved group were all higher before and 6 months after PCI.LPSS (r =-0.578,P ＜0.001) and CPSS (r =-0.817,P ＜0.001) before PCI were both closely related to △LVEF.In single parameter mode of ROC curve analysis,the area under the ROC curve (AUC) (0.867),sensitivity (94.7％) and specificity (74.4％) of CPSS are relatively higher than other STI parameters.In multiple parameters united mode of ROC curve analysis,AUC (0.897),sensitivity (94.7％) and specificity (74.4％) of LPSS,RPSS and CPSS united were the highest among all the combinations of all STI parameters.Conclusions Left ventricular function improvement of patients with AMI 6 months after PCI is accurately assessed and predicted by STI.CPSS is a strong predictor for left ventricular function improvement 6 months after PCI of AMI patients among all the STI parameters and is an effective indicator for the assessment of left ventricular function improvement of AMI patients.

Objective To assess left ventricular function early and late improvement of patients with acute myocardial infarction (AMI) treated by percutaneous coronary intervention (PCI) by speckle tracking imaging (STI).The clinical values of the assessment of STI for the prognosis and heart function improvement of AMI patients treated by PCI were discussed.Methods 73 AMI patients who had AMI for the first time and had been treated by primary PCI from September 2010 to July 2011 and were examined in the follow-ups from December 2010 to February 2012 in our hospital were enrolled.Dynamic images were acquired before PCI,at 3 months and 6 months after PCI and analyzed by STI.Dynamic images were analyzed for longitudinal peak systolic strain (LPSS),radial peak systolic strain (RPSS) and circumferential peak systolic strain (CPSS) values by STI.According to the comparison of left ventricular ejection fraction (LVEF) before PCI and 6 months after PCI,patients were divided into left ventricular function improved group (ΔLVEF6＞5％) and not-improved group.According to the comparison of LVEF before PCI and 3 months after PCI,improved group were divided into left ventricular function early-improved group (ΔLVEF3 ＞5％) and late-improved group.Results The values of all STI parameters before PCI,3 months and 6 months after PCI in improved group were higher than those in not-improved group (P ＜0.001,all).LPSS before PCI and at the follow-ups and RPSS at 3 months after PCI in early-improved group were higher than those in late-improved group (LPSS at 3 months after PCI:P＜0.001;Other parameters:P ＜0.05).There were significant correlations between all STI parameters and both ΔLVEF3 and ΔLVEF6.LPSS before PCI was more closely related to ΔLVEF3 (r =-0.781,P ＜0.001).CPSS at 6 months after PCI was more closely related to ΔLVEF6 (r =-0.834,P ＜ 0.001).Conclusions Early and late function improvement of left ventricle in AMI patients who is treated by PCI are accurately assessed by STI.The precise analyses of longitudinal and circumferential movements in STI are important for clinical diagnosis.

Objective To evaluated the effect of hyperbaric oxygenation therapy(HBO)on the RhoA ex- pression and nerve function after transient focal cerebral ischemia in a rat model of middle cerebral artery occlusion. Methods One hundred and twenty-six healthy Sprague-Dawley rats were used and randomly divided into a sham op- eration group(shame group,n=42),a treatment group(n=42),and a control group(n=42).The animal model of middle cerebral artery occlusion(MCAO)was established by using the Zea-Longa method with the animals in the treatment and the control groups,and sham operation was performed with those in the sham group.HBO was applied to the animals in the treatment group.The RhoA protein expression was observed by using immunohistochemistry technique,and the neurological function was evaluated by Bederson's scale at different time points after MCAO.Re- sults(1)Weakly positive expression of RhoA could be located in bilateral cortex and the basal ganglia in the sham group.The expression of RhoA in the treatment group and control group was increased as early as 6 hours after MCAO when compared with that of the sham group,and peaked at 48 h after MCAO and decreased after then,but was still higher than that of the shame group at 7th day to 14th day after MCAO.It was also found that the expression of RhoA of the treatment group was significantly lower than that of the control group(P

BACKGROUND: Previous studies have confirmed that in the animal models of articular cartilage defects and osteoarthritis, the chondrocytes can overexpress the matrix metal oproteinases. Various abnormal stimuli are likely to break the balance between matrix metal oproteinase and tissue inhibitor of metal oproteinase, thus leading to degeneration of extracel ular matrix of articular cartilage, as wel as the decline and offset of cartilage chondrocytes. OBJECTIVE: To observe the effect of cyclic tensile strain on the expression of matrix metal oproteinases during the repairing process of rabbit articular cartilage defects. METHODS: The animal models of articular cartilage defects were established, and chondrocytes were separated for culture at 10 weeks after operation. The chondrocytes on the non-surgical side were considered as the normal group, and the chondrocytes on the surgical side were randomly divided into high cyclic tensile strain group, low cyclic tensile strains group and control group, and the load amplitude was sin10%. Then 0.1, 1.0 and 0 Hz cyclic tensile strains were loaded respectively. The expressions of matrix metal oproteinases 2, 3, 9 and 13 in each group were detected with reverse transcription-PCR at 24, 48 hours, 1, 2 and 4 weeks after loading cyclic tensile strain. RESULTS AND CONCLUSION: There were significant differences in the expressions of matrix metal oproteinases 2, 3, 9 and 13 at 24 hours after loading cyclic tensile strain between the normal group and the control group (P < 0.05); and there were significant differences in the expressions between the high cyclic tensile strain group and the low cyclic tensile strain group at 1, 2 and 4 weeks after loading cyclic tensile strain (P < 0.05).At the same time, the expressions of matrix metal oproteinases 2, 3, 9 and 13 in the low cyclic tensile strain group were continued to decline, and there were significant differences in the expressions after loading cyclic tensile strain for 24 hours and 4 weeks (P < 0.05). The results indicate that mechanical load can affect the expression of matrix metal oproteinases in the healing process of rabbit articular cartilage defects. In the cel ular and molecular level, the incidence and development of pathological articular cartilage defect and stress should affect each other.

Establishment of the “Happy Reading Club” was to create a personal reading space for undergraduates. The reading situation of undergraduates was analyzed with the creation processes of “Happy Reading Club” summa-rized.The reading popularizing activities were described, including questionnaire investigation of reading, sign of DIY “tree”, and recommendation of classic booklist, with the existing problems pointed out.

Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

To evaluate the automated segmentation algorithm for detection of intra - retinal layers to process images obtained from ultra- high resolution optical coherence tomography ( OCT ) . Graph theory and the shortest path search based on dynamic programming were applied to automatically segment the 8 intra - retinal layers. We experimentally verified the accuracy and reliability of the algorithm. The results showed that the intra-retinal layer boundaries between automated and manual segmentations matched well. The algorithm successfully segmented the intra- retinal layers in glaucoma, high myopia, and retinitis pigmentosa patients. The proposed automatic segmentation for intra-retinal layers provides a promising tool for quantitative analysis in clinical diagnosis and treatment.