Objective To investigate the role of ECRG2 gene in migration and invasion of human fibrosarcoma HT1080 cells. Methods Tet-on system was used to set up ECRG2-inducible cell clones.Knockdown of ECRG2 gene was done by stably expressing short hairpin RNA (shRNA) specific for ECRG2 mRNA in a highly invasive fibrosarcoma cell line HT1080. The expressions of ECRG2 proteins were detected by Western blotting assay. Wound-healing scrape assay and transwell invasion assay were performed to assess the effect of ECRG2 gene on the invasive properties of HT1080 in vitro. Results The most effective ECRG2siRNA interference and inducible was selected by Western blotting assay. Knockdown of ECRG2 gene significantly promoted the migration and invasion of HT1080 cells while expression of ECRG2 impaired the migration and invasion of HT1080 cells in vitro. Conclusion ECRG2 gene is correlated with the invasive and metastatic potential of fibrosarcoma carcinoma, and expression of ECRG2 induces the inhibition of migration and invasion of HT1080 cell line.

BACKGROUND: Studies showed cellular adhesion molecule and neurotrophic factor secreted from olfactory ensheathing cells (OECs) could protect the spinal neurons and promote the regeneration of spinal axon. OBJECTIVE: To compare the competence to repair spinal cord injury between olfactory mucosa OECs and olfactory bulb OECs. DESIGN, TIME AND SETTING: Randomized control animal experiment was performed in the central laboratory of Xidian Group Hospital between June 2007 and June 2008. MATERIALS: Twelve male SD rats were randomized selected and divided into experiment group (n=6, 23 months old) and control group (n=6, 3 months old). They were used for in vitro culture and purification of OECs; other 30 SD rats were randomized into three groups of 10 rats each: neonatal rat olfactory bulb OECs transplantation group, normal olfactory mucosa OECs transplantation group and blank control group.METHODS: Spinal cord injury models were produced in 30 rats, which were transplanted with the neonatal rat or SD rat OECs cultured in vitro. No transplant was given in the control group. MAIN OUTCOME MEASURES: At 4 and 8 weeks postoperation, the Basso, Beattie, Bresnahan (BBB) score for nerve function, the evoked potential of legs and the histopathological diversify of injured spinal cord. RESULTS: Seven rats died dudng the experiment process, and the death rate was similar between groups. At 4 and 8 weeks postoperation, there was no significant difference in the BBB scores between neonatal rat olfactory bulb OECs transplantation group and normal olfactory mucosa OECs transplantation group (P > 0.05), which were both significantly higher than blank control group (P < 0.001); the BBB scores in two transplantation groups were higher at 8 weeks than at 4 weeks (P < 0.01 ). At 4 weeks postoperaUon, no animal was shown to elicit motion evoked potential, but it was present in two transplantation groups at 8 weeks, with no significant difference between two groups (P > 0.05). The blank control group had still no motion evoked potential (P < 0.001 ). At 8 weeks postoperation, more cell infiltrations were found in the injured spinal cord of two transplantation groups, while few in the control group.CONCLUSION: Both OECs dissociated from olfactory bulb and olfactory mucosa have the same ability to repair the injured spinal cord, and their effect is similar.

At present,reports at home and abroad suggest a low probability of successful preservation of ovarian endocrine function after ovarian shift radiotherapy.After radiotherapy for cervical carcinoma,the ovarian function is associated with various factors,such as radiotherapy dose and method,patient's age,shift position,and concurrent chemotherapy drugs.Therefore,as for each patient,the dose to the ovarian tissue should be controlled within the individual dose limit to effectively preserve the ovarian function.

Objective To discuss the value of mammotome biopsy of breast cakified lesions under the real time guidance of mammography.Methods The data of 42 patients with 44 mammary gland calcification(2 patients with bilateral lesions)who underwent mammotome biopsy under the real time guidance of a mammography from Jan.2011 to Feb.2012 in the First Affiliated Hospital of Chongqing Medical University were retrospective analyzed.Rapid paraffin pathology was conducted.Results Among the 44 lesions,34 were benign(27 hyperplasia,2 fibroadenoma,1 lipoma,1 cyst,3 others)and 10 were breast cancer(a benign lesion diagnosed as breast cancer by postoperative biopsy was included).Among the 42 patients,32 patients had benign lesions,accounting for 76.2％ (32/42),10 patients had breast cancer,accounting for 23.8％ (10/42).All lesions were completely resected during the operation.Malignant lesions were given treatment such as modified radical mastectomy,breast conservation surgergery,chemotherapy and so on.A single lesion resection took about 20 minutes.During the follow-up of 3 months,no significant event happened.Conclusions Through the qualitative diagnosis of mammotome biopsy under the real time guidance of mammography,we can get early diagnosis of breast cancer.Early diagnosis and early treatment can improve the life quality and survival time.It is more important to detect early breast cancer which was considered benign lesions clinically.Mammotome biopsy under the real time guidance of mammography has several advantages including easy to operate,accurate positioning and less complications,making it the preferred approach in managing mammary gland calcification especially suspicious malignant calcification.

Objective To investigate the role of ECRG2,a novel tumor suppressor gene,in spindle assembly checkpoint. Methods Using siRNA approach to deplete the expression of ECRG2, using immunofluorescence to test the distribution of ECRG2,using Western blotting to examine the expression cell cycle proteins.Results ECRG2 localized to centrosomes during interphase and kinetochores during mitosis.Further analysis revealed that ECRG2 participates in the spindle assembly checkpoint.Depletion of ECRG2 abolished the spindle assembly checkpoint.Conclusion Our results indicated that ECRG2 is important for ensuring spindle assembly checkpoint,accurate chromosome segregation,and its depletion may contribute to chmmosome instability and aneuploidy in human cancers.

Objective: To examine the effects of ? carotene and ? tocopherol on LDL oxidation by monocyte derived macrophages from SD rats (in DMEM medium). [WT5FZ]Methods: Macrophages were incubated with LDL in DMEM in which freshly dissoved VE (40,100,200 ?mol/L) and ?C (0.5,1.0,2.0 ?mol/L) had been added. After incubating for 24 h, the mediums were centrifuged to determine TBARS and lipofusin and dienes. [WT5FZ]Results: Supplementation of ? tocopherol significantly decreased TBARS, lipofusin and conjugated dienes production and electrophoretic mobility. Supplementation of ? carotene at 0.5 ?mol/L significantly decreased TBARS, lipofusin and conjugated dienes production and electrophoretic mobility, but other levels showed no effect. Increasing ? carotene concentration to 2.0 ?mol/L in the system increased the lipofusin production compared with control group. ? tocopherol at 200 ?mol/L and ? carotene at 0.5 ?mol/L were the most effective in all dose groups. [WT5FZ]Conclusion: Cell mediated oxidation of LDL can be regulated by antioxidants such as ? tocopherol and ? carotene.

Objective Using the effect of microneedle array to investigate the feature and rule of transdermal drug delivery of sinomenine hydrogel patch.Methods The microneedles with different length and sinomenine hydrogel patch were prepared;The isolated rat skin was pretreated by different needle-shapes of microneedle array and different timepoints with the same needle-shape,and then sticked the sinomenine patch.The permeation rates of sinomenine were studied using a Franz diffusion cell and compared with passive diffusion of the rat skin untreated.Sinomenine content was measured by HPLC.Results The permeation rates of sinomenine via the skin pretreated with 100 and 200 ?m microneedles were 40.7 and 52.4 times to those of the untreated.The permeation rates of sinomenine via the skin pretreated 7 min with 200 ?m microneedles is 142.0 times to those of the untreated.When skins were pretreated using 200 ?m microneedles with different force, the drug permeation rates were increased with the force increasing.When the force exceeded 5 N,the drug permeation rates were equilibrium.Conclusion When transdermal drug delivery by using microneedle array-hydrogel patch,the permeation rates increase significantly.The needle-shape,skin pretreating time,and force of the microneedles play the important roles in the transdermal drug delivery.

AIM To establish the expermental model in vitro for the study of mechanism of antitumor drug resistance and the screening of antitumor drug. METHODS By continuously exposing cells to gradually increasing concentration of drug and agar cell colony forming technique, using dye exclusive method for determing cytotoxic effect, a murine leukemia L1210 cell subline were established,which exhibited 40 fold resistance to Cis diamminedichloro platinum(DDP). RESULTS The doubling times, plate efficiency, cell cycle, DNA index and cell morphology of DDP resistant L1210 subline were similarto those of its parant cell line. When the cell subline was stored with DDP at -196℃ for 6 months and then was recovered, its characterization of antitumor drug resistance was still maintained to a period of 5 months without DDP. DDP resistant L1210 subline was characterized with cross resistance to Carboplatin, Mitomycin, Thio Tepa, Methotrexate, Vincristin and Mustine Hydrochloride, but with no cross resistance to Harringtonine and Adriamycin. It was seemed more sensitive to Cytarabine and Fluorouracil. CONCLUSION DDP resistant L1210 subline is a good experimental model in vitro for the study of mechanism of antitumor drug resistance and the screening of antitumor drug.

Aim To research the effects of total flavones of rhododendra(TFR)on transient receptor potential vanilloid receptor 4(TRPV4)in cerebral basilar arteries(CBA)of rats subjected to ischemia/reperfusion(IR)injury.Methods The model of total brain IR was established by four-artery occlusion(4-VO)method in rats.Arterial pressure perfusion and cell membrane potential recording methods were used for surveying the dilatation and hyperpolarization of TFR and ruthenium red(RR,an inhibitor of TRPV4)in the KCl-preconstricted CBA ex vivo in rats subjected to IR.Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot were utilized to investigate the TRPV4 mRNA and protein expressions of TFR and RR in cerebrovascular endothelial cells of CBA in vivo in rats subjected to IR.Results 11～2 700 mg·L-1 TFR significantly induced concentration-dependent hyperpolarization and dilatation in the KCl-preconstricted CBA in rats subjected to IR.TFR still produced degenerative hyperpolarization and dilatation by removal of endothelium in CBA,which was remarkably attenuated as compared with endothelium-intact group(P<0.01).After removal of NO and PGI2 vasodilatation,TFR obviously elicited the hyperpolarization and dilatation that were further decreased by RR(an inhibitor of TRPV4)in IR CBAs.TFR pretreatment apparently increased the level of TRPV4 mRNA and protein expressions in IR CBAs.These effects were restrained by RR,an inhibitor of TRPV4.Conclusions TFR could mediate endothelium-dependent and endothelium-independent effects.The endothelium-derived dilatation may be related to the increase of endothelium activity and endothelium-derived hyperpolarizing factor(EDHF)generation and release that have been promoted by TFR,and secondarily activating TRPV4,which results in Ca2+ inflow and subsequent hyperpolarization of vascular smooth muscle cell membrane and vasorelaxation.

Objective To assess the variation in patient setup corrections for three different anatomic treat-ment sites using daily pretreatment megavoltage CT(MVCT)in helical tomotherapy,and to analyze alternative refer-ence margins for specific tumor site.Methods Sixty patients treated for three anatomical sites on helical tomotherapy were analyzed.Daily MVCT was carried out for registration and setup corrections before each treatment fraction.Setup errors and rotational setup corrections from 587 head and neck,500 thoracic,371 abdomen and gynecology MCVT imaging were input to statistical analysis.Incidences of three dimensional vector error lengths were calculated for each anatomic site.Overall distribution histograms of the three -dimensionally error was presented using GraphPad Prism 5.The distributions of systematic and random setup errors were quantitative analyzed and the additional margins required were also taken into account.Results 1 458 MVCT scans were implemented for sixty patients.Head and neck had lower frequencies of translational setup errors than others.Frequency of at least 3mm three -dimensional setup errors for head and neck,thoracic,abdomen and gynecology was 55.3%,70.8%,79.8%,respectively.This fre-quency decreased to 17.5%,40.6%,47.2% if 3D vector distance ≥5mm was scored.Overall systematic errors ranged from -1.4mm to 2.7mm,abdomen and gynecology had the largest setup errors in the vertical direction which was statistically significant(χ2 =19.3,P <0.05).The suggested margins should be increased by 4 -7 mm in three -dimensional direction for head and neck,9 -14mm for thoracic,10 -17mm for abdomen and gynecology.Conclusion Differences in setup corrections are perceived between head and neck,thoracic,abdomen and gynecology.The accura-cy of patient positioning can be improved if pre -treatment daily MVCT scans are put into use.Results from setup cor-rection can provide evidence for tumor treatment margin and improve the accuracy of regular radiotherapy.