Objective To evaluate the diagnostic evaluation of isotropic scanning and Lung Care soft- ware in solitary pulmonary nodules,and to improve the diagnostic accuracy.Methods 52 patients suffered from SPN were included in our study.Two experts in CT analyzed the films.First,they read the axial images and made diagnosis.Then isotropic scanning and lung care software approaches were used on 16 spiral CT and another analysis were made again.The results were compared with pathological diagnosis respectively. Results Spiculated sign,lobulated sign,vessel convergence were found more on isotropic scanning approach, that had significant difference with axial images analysis(P

Background Proliferation of the human Tenon capsule fibroblasts(HTFs) is a main cause of failure of filtering surgery.To search the drug of inhibiting the growth of the HTFs is essential for the improvement of successful rate of filtering surgery.Objective The aim of this study was to investigate the inhibitory effect of apigenin on HTFs and its mechanism.Methods Human Tenon capsular tissue was obtained during the strabismus correction surgery.HTFs was primarily cultured using explant method and identified using vimentin by immunochemistry.The 3-5 generation of cells were incubated to 96-well plate.Apigenin of 0,20,40,80,160 μmol/L was added into the medium,respectively,for 24,48,72 hours,and the proliferation of HTFs was detected by sulfonyl chloride (SRB) at the wavelength of 560 nm (A560).Bromodeoxyuridine (BrdU) of 10 μg/L was added to culture the cells for 48 hours to calculate the labeling rate of BrdU.The morphology of the cells was observed using Hoechst 33258 staining,and apoptosis and cells cycle were evaluated by flow cytometry.Results Cultured cells grew well with the positive response for vimentin,showing the green fluorescence in cytoplasm.SRB assay showed that the A560 value was gradually declined with the increase of the dosage of apigenin and prolong of time (Fgroup =480.306,P =0.000 ; Ftime =555.144,P =0.000).The labeling rate after 0,40,80 μmol/L apigenin acted for 48 hours was (87.860 ±0.632)％,(61.520±4.306)％ and (23.480±4.472)％,showing a significant difference among the three groups (F =299.347,P =0.000).The labeling rate of HTFs for BrdU was significantly decreased in the 40 and 80 μmol/L apigenin groups compared with the 0 μmol/L apigenin group (P＜0.05).Hoechse 33258 staining found that the number of the HTFs was gradually decreased and the cell number of karyopyknosis and nuclear deformation was increased with the increase of apigenin dosage.Percentage of cells in G0/G1 phase were raised and that in S and G2/M phase were declined in the higher dosage apigenin group,with a significant difference among the different groups (FG0/G1 =58.621,P=0.000;Fs =32.357,P=0.001 ;FG2/M =83.998,P=0.000).In the 72nd hour after acted by 0,40,80,160 μmol/L apigenin,the apoptosis rate of HTFs was (4.77±0.21) ％,(13.24±1.35)％,(18.33±1.86) ％,(31.58 ± 2.77) ％,respectively,with a statistically significant difference among the four groups (F =204.791,P＜0.05).Conclusions Apigenin restrains the growth of HTFs by evoking G0/G1 cell cycle arrest and inducing apoptosis in a dosage-and time-dependent manner.

Neural stem cells in brains have capacities of proliferation and differentiation, which is very critical to rebuild the cerebral cortex functions. Therefore, it is of great importance to find key targets and network pathways that regulate the proliferation of neural stem cells, which is also a pressing problem in the medical circle. With the Notch pathway as the core of the network, this paper summarized the advance of the bimolecular network system composed of Wnt, Shh, EGFR, cytokines and Notch signal, and analyzed such key nodes as Notch receptor, CBF1, NICD, Hesl, which may become potential targets of new-type drugs in the future. With the multi-component, multi-target, multi-lever characteristics, traditional Chinese medicines have many common grounds with the network pharmacology. The active component groups or active ingredients in traditional Chinese medicines are one of the material bases for showing their network pharmacological effect, which is worth exploring. This paper aims to provide a new strategy for the treatment of neurodegenerative disease and nerve injury with traditional Chinese medicines.
Animals ; Cell Proliferation ; Humans ; Neural Stem Cells ; cytology ; metabolism ; Signal Transduction ; Systems Biology

Objective To study the process and mechanism underlying Wallerian degeneration of the central nervous system after stroke.Methods A case suspected of stroke with bilateral symmetrical lesions in middle cerebellar peduncles (MCPs) was described.The etiology of bilateral MCP abnormal signals on MR was analyzed according to the clinical process and neuroanatomy.Results Unilateral paramedian pontine infarction,covering the crossing area of pontocerebellar fibers, would cause bilateral secondary degeneration of MCPs,with hyperintense signals on T2-,Flair and diffusion-weighted images.Conclusions Wallerian degeneration of projecting system is a common sequel after stroke and should not be misdiagnosed as other diseases.

Objective To analyze and compare the clinical application values of procalcitonin (PCT),leukocyte count (WBC) and C-reactive protein(CRP) in elder patients with infection.Methods In patients(age≥ 65 yrs,axillary temperature ＞38.0℃)with infection or suspected infection,PCT,WBC,CRP and other bacteriological examination were performed.The electronic medical records from the HIS system of our hospital were analyzed retrospectively in combination with medical history.Results Of the enrolled 219 patients,65 ones were in control group,48 ones SIRS,51 ones sepsis and 55 ones MODS.There was a positive correlation between the level of serum PCT and the infection degree.The Spearman correlation coefficient was 0.706 (95％CI:0.616-0.797,P=0.000).Based on the highest Youden index (sensitivity+specificity-1),the best cutoff point of diagnosis for PCT was ＞0.341 μg/L (sensitivity 84.5％,specificity 55.8％),a analysis of receiver operating characteristic(ROC) curve about PCT,WBC and CRP was carried.Area under the curve (AUC) of PCT to controlled infection was 0.916 (95％CI:0.864-0.967,P=0.000).Based on the highest Youden index (sensitivity+ specificity-1),the judging threshold of PCT to infection controlled or not was 0.73 μg/L (sensitivity 84.6％,specificity 88.0％).PCT level after treatment ＞0.73 μg/L showed the uncontrolled infection,＜ 0.73 μg/L controlled.Conclusions PCT has a higher specificity for elder patients with infection.The variation of PCT level can guide the application of antibiotics,avoid abuse and decrease the occurrence of drug-resistant bacteria.

Objective To investigate clinical features and prognostic factors in acquired immune deficiency syndrome (AIDS) patients with opportunistic infections.Methods Forty-two cases of AIDS patients with opportunistic infections were enrolled in this study.Clinical data and major factors affecting the prognosis were analyzed using Logistic regression.Results Bacterial infection was the first etiological factor(57.1%) of opportunistic infections in AIDS patients.Eighty-three point three percent of patients infected with more than two kind of etiological agents.Fifty-seven point one per- cent of cases were infected in multiple sites.CD4~+ T cells count was associated with the opportunistic infections.Conclusions The CD4~- T lymphocytes count is the key factor affecting the prognosis of AIDS patients with opportunistic infections.The average of CD4~+ T lymphocytes count is significant- ly related with the major opportunistic infections in AIDS paitents.

Objective To investigate the effects of low frequency ultrasound (LFU) on the proliferation and apoptosis of smooth muscle cells (SMCs) in rabbits with carotid atherosclerosis(CA).Methods CA models were established in 30 New Zealand rabbits using a high fat diet and air-drying.They were randomly divided into a control group and four LFU groups:group A received 0.5 W/cm~2 LFU for 5 min/d,group B 0.5 W/cm~2 for 10 min/d, group C 1 W/cm~2,5 min/d,and group D 1 W/cm~2,10 min/d.The rabbits' carotid arteries were autopsied after 20 d of the LFU treatment.The expression of PCNA and TUNEL staining were used to explore the proliferation and apopto- sis of SMCs,and the proliferation rates (PRs) and apoptosis rates (ARs) were calculated.Results Compared with the control group,the PRs in groups B,C and D were significantly lower,while the ARs in groups B,C and D were significantly higher.There was no significant difference in ARs or PRs among groups B,C and D.Con- clusion LFU can induce SMC apoptosis and inhibit SMC proliferation in rabbits with CA.

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

BACKGROUND:Retinoid X receptor (RXR) plays a central role in the regulation of intracellular receptor signaling pathways. The activation of RXR has protective effect on H2O2-induced apoptosis of H9c2 ventricular cells in rats. But the protective effect and mechanism of activating RXR in cardiomyocytes against hypoxia/reoxygenation (H/R)-induced oxidative iniury are stillunclear. METHODS:The model of H/R injury was established through hypoxia for 2 hours and reoxygenation for 4 hours in H9c2 cardiomyocytes of rats. 9-cis-retinoic acid (9-cis RA) was obtained as an RXR agonist, and HX531 as an RXR antagonist. Cultured cardiomyocytes were randomly divided into four groups:sham group, H/R group, H/R+9-cis RA -pretreated group (100 nmol/L 9-cis RA), and H/R+9-cis RA+HX531-pretreated group (2.5 μmol/L HX531). The cellviability was measured by MTT, apoptosis rate of cardiomyocytes by flow cytometry analysis, and mitochondrial membrane potential (ΔΨm) by JC-1 fluorescent probe, and protein expressions of Bcl-2, Bax and cleaved caspase-9 with Western blotting. Allmeasurement data were expressed as mean±standard deviation, and analyzed using one-way ANOVA and the Dunnett test. Differences were considered significant whenP was <0.05. RESULTS:Pretreatment with RXR agonist enhanced cellviability, reduced apoptosis ratio, and stabled ΔΨm. Dot blotting experiments showed that under H/R stress conditions, Bcl-2 protein level decreased, while Bax and cleaved caspase-9 were increased. 9-cis RA administration before H/R stress prevented these effects, but the protective effects of activating RXR on cardiomyocytes against H/R induced oxidative injury were abolished when pretreated with RXR pan-antagonist HX531. CONCLUSION:The activation of RXR has protective effects against H/R injury in H9c2 cardiomyocytes of rats through attenuating signaling pathway of mitochondria apoptosis.

OBJECTIVETo investigate the effects of 50 Hz magnetic fields (MF) on DNA double-strand breaks in human lens epithelial cells (hLECs).

METHODSThe cultured human lens epithelial cells were exposed to 0.4 mT 50 Hz MF for 2 h, 6 h, 12 h, 24 h and 48 h. Cells exposed to 4-nitroquinoline-1-oxide, a DNA damage agent, at a final concentration of 0.1 micromol/L for 1 h were used as positive controls.After exposure, cells were fixed with 4 % paraformaldehyde and for H2AX (gamma H2AX) immunofluorescence measurement. gamma H2AX foci were detected at least 200 cells for each sample. Cells were classified as positive when more than three foci per cell were observed. Mean values of foci per cell and percentage of foci positive cells were adopted as indexes of DNA double-strand breaks.

RESULTThe mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 24 h were (2.93 +/-0.43) and (27.88 +/-2.59)%, respectively, which were significantly higher than those of sham-exposure group [(1.77 +/-0.37) and (19.38+/-2.70)%, P <0.05], and the mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 48 h were (3.14 +/-0.35) and (31.00 +/-3.44)%, which were significantly higher than those of sham-exposure group (P <0.01). However there was no significant difference between 50 Hz MF exposure groups for 2 h, 6 h, 12 h and sham-exposure group for above two indexes (P >0.05).

CONCLUSION0.4 mT 50 Hz MF exposure for longer duration might induce DNA double-strand breaks in human lens epithelial cells in vitro.

Cells, Cultured ; DNA ; radiation effects ; DNA Breaks, Double-Stranded ; radiation effects ; DNA Damage ; radiation effects ; DNA Repair ; radiation effects ; Electromagnetic Fields ; Epithelial Cells ; metabolism ; radiation effects ; Humans ; Lens, Crystalline ; cytology