Objective　To study the occurence, development and regulation of reactive gliosis with astrocyte (Ast) in vitro. Methods　Ast was isolated and cultured in vitro and its model of reactive gliosis was established by scratching the cultured astrocytes. The reactivity and rules of Ast to injury was studied by morphological changes, RT-PCR, immunocytochemistry, in situ hybridization and imaging analysis. Results　After scratching, the astrocytes showed typical features of reactive gliosis, with the hypertrophic cell body, thickened and lengtheded processes, and enhanced glial fibrillary acidic protein (GFAP) staining. In situ hybridization and RT-PCR analysis confirmed that the expression of GFAP mRNA was markedly increased. These changes occurred 1 d after scratching and reached the peak 5 to 7 d after injuring. Conclusion　A model of reactive astrogliosis was successfully established in vitro which showed an active reaction to injury. The characteristics of reactive gliosis parallel that seen in vivo.

The changes of the morphology and percentage of the macroglia surrounding focus of brain stabbing injury (BSI) were observed by immunohistochemistry,immunoinfluorescence stain and flow cytometer(FCM) and the effect of magroglia during glial scar forming were elucidated.The results showed that a large number of GFAP-immunoreactive positive cells were accumulated around the focus of BSI.These cells were hyperplastic,hypertrophic,and emerged swollen cytoplasmic processes.The most marked changes were observed at 1-2 week after BSI.The results of FCM showed that the percentage of GFAP positive cells increased gradually and reached to a peak during 1～2 week after BSI.The peak ratio of GFAP positive was about 46%.However,the changes of morphology and number of GC positive cells were not detected after BSI.The authors believed that astrocyte is the main macroglia during glial scar formatting .The oligodendrocytes is not an active cell during this course.

Objective To investigate the effects of mutated PS-1 on the proliferation and apoptosis of RA-induced PC12 cells . Methods The MTT assay, flow cytometry, TUNEL method were used in the research. Results An inhibition of proliferation was demonstrated in the RA-induuced PC12 cells expressing PS-1 mutant L286V. The proliferation of the cells was blocked in the G1→S phase. Whether with or without fetal bovine serum(FBS), PS-1 mutant L286V was able to accelerate the apoptosis of RA-induced PC12 cells. This action was enhanced markedly under the culture without FBS. Conclusions The RA-induced PC12 cells expressing PS-1 mutant L286V shows that the proliferation is inhibited and blocked in the G1→S phase. Whether with or without FBS, PS-1 mutant L286V can accelerate the apoptosis of RA-induced PC12 cells. This action is enhanced markedly under the culture without FBS.

Objective To study the relationship between GFAP and the malignant degree of astrocytoma. Methods After successful transfer of retrovirus with GFAP (PLBsKG) into C6 cells mediated with liposome, the morphology, growth curve and the proliferative cycle of the target cells were observed. The expression of the protein and mRNA of GFAP was determined with RT-PCR and immunocytochemical staining. In addition, the relationship between the staining GFAP-IR and the malignant degree of astrocytoma was analyzed according to Kermohan′s grading standard in 20 samples of the cells of astrocytoma with imaging analytic system. Results PLBskG resulted in the decrease of GFAP in the target C6 cells, and changed the morphology and increased the proliferation of C6 cells. The number of the cells in the G 2+M phages was increased. The intensity of GFAP-IR staining was negatively correlated to the malignant potency of astrocytoma. The more intensive staining of the cells, the lower of the malignant degree of astrocytoma and vice versa. Conclusion There is a close relationship between GFAP and malignant degree of astrocytoma. The level of GFAP expression serves the index of the malignant degree and prognosis of astrocytoma in certain degree.

Objective To explore the clinical value of laparoscopic-assisted vaginal hysterectomy(LAVH)in large uterus.Methods Retrospective analysis was conducted on clinical data of 94 patients(whose uterus were as big as 10-18 gestational weeks)who received hysterectomy from January 2005 to March 2007,in which 56 cases were performed laparoscopic-assisted vaginal hysterectomy(LAVH group)and 38 cases vaginal hysterectomy(VH group).The operation time,blood loss,postoperative hospital stay,and the incidence of postoperative complications were compared between the two groups.Results Compared with VH group,there were a lower chance of abdominal hysterectomy(0/56 vs 5/38,?2=5.389,P=0.020),a shorter operation time [(149?11)min vs(179?14)min,t=-11.610,P=0.000] and a shorter postoperative hospital stay [(5.8?1.4)d vs(7.3?3.6)d,t=-2.825,P=0.006] in the LAVH group.There were no significant differences in blood loss,morbidity and time to first flatus between the two groups.Conclusions The LAVH extends the indications of VH,ensuring the safety of VH for the uterus bigger than 10 gestational weeks,therefore it is an operative procedure to be recommended.

Objective To investigate the cholinergic neuron properties and the changes of proliferation and apoptosis of PC 12 cell induced by different concentrations of all trans retinoic acid(RA). Methods PC 12 cell was induced by RA of concentrations of 1-50 ?mol/L. Changes of cell growth curve and cycle, apoptosis and changes of activities of acetylcholinesterase(AchE) and choline acyltransferase(ChAT) were detected by MTT assay, flow cytometry, TUNEL method, spectrophotometry and radiochemistry assay, respectively. Results ① RA significantly enhanced the activity of ChAT with concentrations between 1 to 20 ?mol/L. The activity of ChAT with concentration of 10 ?mol/L was higher than those with other concentrations. ② The apoptosis of RA induced PC 12 cells was markedly enhanced when concentration was higher than 10 ?mol/L. ③ A concentration dependent inhibition of proliferation was demonstrated in the RA induced PC 12 cells. ④ PC 12 cells proliferation was blocked in the G 1→S phase. Conclusion RA can be used to induce PC 12 cells to show the properties of cholinergic neuron with an optimal concentration of 10 ?mol/L.

Objective In order to explore the ways of reflecting the dose distribution in the implementation of the of IMRT (intensity modulated radiation therapy),a 2D diode array (2D-DA) was used in verifying the composite dose distribution of IMRT plans in the way of multi-gantry-angle composite (MGAC).Methods IMRT quality assure (QA) plans of 27 patients,based on the 2D-DA and solid water phantom,were designed and verified in two ways of single-gantry-angle composite (SGAC) and MGAC verifications.The comparison and analyzation of the dose distributions of the TPS calculation and the measurement of the 2D-DA were done.Results (1) When the beam central axes were not superposed with the detectors'plane of the 2D-DA,the verification passrate of SGAC and MGAC planar dose distribution of 27 patients'IMRT plan were 94.56%±4.28% and 94.81%±3.80% (the criteria:rvalue,3 ram/3%),respectively.There was no statistical difference between the results of two sets (t =-0.213,P＞0.05).(2) When one of the beam central axes was superposed with the detectors'plane of the 2D-DA,the verification passrate of MGAC planar dose distribution were 79.72%±12.77%.Conclusions Using the 2D-DA with a proper phantom,there was no statistical difference in the SGAC and MGAC verifications of IMRT plans when the beam central axes were not superposed with the detectors'plane.However,the MGAC dose distribution can provide more about the clinical dosimetry,and the errors in the implementation of the of IMRT were easier located.

Objective To study the regulatory effect of ethanol extract of glossy privet fruit and its monomer tyrosol on the adhesion and migration of human epidermal melanocytes.Methods Epidermal melanocytes were isolated from human foreskin,and subjected to a primary culture.Mter 3-5 passages,the melanocytes were treated with various concentrations of ethanol extract of glossy privet fruit (0.0375-0.6 mg/ml)and tyrosol (0.125-2 mmol/L) for 24-72 hours.The XTT colorimetric assay was carried out to evaluate the proliferation of melanocytes,fibronectin (FN)-coated culture plates were used to evaluate the adhesion activity of melanocytes,and Transwell assay was conducted to assess the migration activity of melanocytes.Confocal laser microscopy was utilized to observe the structure and distribution of actin cytoskeleton in melanocytes,and cellular fluorescence intensity was determined by a semi-quantitative analysis.Statistical analysis was carried out by using unpaired t test.Results The adhesion activity of melanocytes to FN was significantly enhanced by the ethanol extract of 0.0375-0.6 mg/ml from glossy privet fruit (P ＜ 0.05 or 0.01),and by tyrosol of 0.5-2 mmol/L (P ＜ 0.05 or 0.01).As XTT assay showed,neither the ethanol extract of 0.15 mg/ml nor tyrosol of 2 mmol/L had cytotoxicity or promotive effect on cell proliferation.Hence,0.15 mg/ml and 2 mmol/L were determined as the working concentrations of ethanol extract and tyrosol respectively.The number of cells migrating through micropore membranes per high-power field (× 200) was 43.7 and 51.0 in melanocytes treated with the ethanol extract of 0.15 mg/ml and tyrosol of 2 mmol/L,respectively,significantly higher than that in untreated melanocytes (20.3,both P ＜ 0.01).Compared with untreated melanocytes,those treated with the ethanol extract of 0.15 mg/ml and those with tyrosol of 2 mmol/L showed higher intracellular fluorescence intensity (P ＜ 0.01) and more stress fiber bundles which congregated inside the cell membrane and around the nuclei.Conclusions The ethanol extract of glossy privet fruit and its monomer tyrosol can promote the adhesion and migration of human melanocytes in vitro,likely by promoting the congregation of actin cytoskeleton in melanocytes.

Objective To analyze the difference of prescription dose between ICRU report 83 and Chinese recommendation in the nasopharyngeal carcinoma (NPC) for intensity modulated radiation therapy (IMRT).Methods Eighty-four NPC were treated using IMRT technology from Jan 1,2010 to Apr 1,2012.All dose volume histogram of the 84 IMRT plan were analyzed retrospectively.The target volumes of planning gross tumor volume of nasopharynx (PGTVnx) or planning clinical target volume and high risk lymphatic drainage (PCTV1) and doses of D100,D98,D95,D50,D2 and D0 were recorded.The mean,standard error,medial,range,coefficient of variation (CV) of PGTVnx,PCTV1,and D100,D98,D95,D50,D2and D0 were calculated,respectively.The homogeneity index (HI) and deviation between D95 and D50 of PGTVnx and PCTV1 were calculated,respectively.The differentiation of grouping results were analyzed with grouped t-test method.Results The HI of PGTVnx and PCTV1 were 0.118 ± 0.045 and 0.272 ± 0.037,respectively.It is the bigger target volume,the worse HI;and the advanced T stage,the worse HI.Either PGTVnx or PCTV1,D95 were less than D50.The average deviation was-5.15％ and-10.97％,and the actual difference value was (382± 180) cGy (P=0.000) and (741± 140) cGy (P=0.000).Conclusions D550,which is the recommendation prescription dose of PTV in ICRU report 83,could evaluate accurately the IMRT plan with combining D98 and D2· When D50 is used to instand of D95,the prescription dose of PGTVnx and PCTV1 should increase 5％ and 11％,respectively.

Objective To evaluate the effect of tacalcitol on the proliferation,adhesion,migration and c-kit mRNA expression of cultured human epidermal melanocytes.Methods Cultured epidermal melanocytes from the prepuce of adolescent males were treated with various concentrations of tacalcitol.Then,cell proliferation was evaluated by tetrazolium salt (XTT) assay after 24,48 and 72 hours of treatment,adhesive activity by using fibronectin-coated culture plates after 72 hours,migratory activity by Transwell assay using a microporous membrane after 24 hours,and the c-kit mRNA expression was semiquantitatively analyzed by reverse transcription PCR after 72 hours of treatment.Statistical analysis was done by repeated-measure analysis of variance and completely random design analysis of variance.Results As repeated-measure analysis of variance showed,tacalcitol of 10-10,10-9,10-8,10-7 and 10-6 mol/L significantly promoted the proliferation of melanocytes (F =9.47,P ＜ 0.01),with significant differences in the promoting effect among various durations of treatment with different concentrations of tacalcitol (F =14.44,P ＜ 0.01),and with significant interaction effect between drug concentration and treatment duration (F =2.47,P ＜ 0.01).The highest proliferation level was observed in melanocytes treated with tacalcitol of 10-s mol/Lfor 72 hours.There was a significant increase in the adhesion rate of human epidermal melanocytes to fibronectin after treatment with tacalcitol of 10-8-10-7 mol/L for 72 hours (both P ＜ 0.01),number of melanocytes migrating through micropore membranes per high-power field (× 200) after treatment with tacalcitol of 10-9-10-8 mol/L for 24 hours (both P ＜ 0.01),and in the c-kit mRNA expression in melanocytes treated with tacalcitol of 10-9-10-7mol/L for 72 hours (all P ＜ 0.01).Conclusion Tacalcitol can promote melanocytes to proliferate,migrate,express c-kit mRNA,and adhere to fibronectin.