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 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 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 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 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.

This study examined the effect of astilbin on the proliferation of rat aortic smooth muscle cells (RASMCs) induced by angiotensin II (AngII) and explored the possible mechanisms. Cell proliferation model of RASMCs was induced by treatmente with AngII. Cells were randomly divided to 8 groups. Normally cultured VSMCs serves as blank control group; in AngII model group, cells were treated with AngII at 10(-7) mol/L; in three astilbin groups, cells were treated with 10, 15, 30 mg/L of astilbin; in three AngII+astilbin groups, cells were treated with AngII (at 10(-7) mol/L) and astilbin at 10, 15, 30 mg/L. Cell proliferation ability was detected by MTT method and the cell cycles and proliferation index were flow cytometrically determined. The expression of c-myc mRNA was assessed by using reverse transcription polymerase chain reaction (RT-PCR), and the expression of NF-κB in RASMCs was immunocytochemically observed. Our results showed that MTT metabolism in RASMCs in the basic and AngII stimulated situation was inhibited by astilbin, and the cells numbers of G(0)/G(1) phase were increased and that of G(2)/S phase were decreased markedly. Not only highly expression of c-myc gene stimulated by AngII could be inhibited by Astilbin significantly, but also the expression of NF-κB protein can be down regulated by Astilbin. We are led to conclude that astilbin astilbin can inhibit the AngII-mediated proliferation of RASMCs by blocking the transition of RASMCs from G(0)/G(1) phase to S phase and by down-regulating the expression of NF-κB, c-myc gene.

In order to study the effect of nitric oxide (NO) on the expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) mRNA in hypoxic pulmonary hypertension (HPH) rats, 30 healthy male Wistar rats were randomly divided into normoxic control group, chronic hypoxic group and hypoxia plus L-arginine (L-Arg) group. The animal model of HPH was developed. The mean pulmonary arterial pressure (mPAP) was measured by inserting a microcatheter into the pulmonary artery. The HIF-1alpha mRNA expression levels were detected by in situ hybridization (ISH) and semiquantitative RT-PCR. It was found that after 14 days hypoxia, the mPAP in normoxic control group (17.6 +/- 2.7 mmHg, 1 mmHg=0.133 kPa) was significantly lower than that in chronic hypoxic group (35.8 +/- 6.1 mmHg, t=0.2918, P<0.05) and mPAP in chronic hypoxic group was higher than that in hypoxia plus L-arginine group (24.4 +/- 3.8 mmHg, t=0.2563, P<0.05). ISH showed that the expression of HIF-1alpha mRNA in the intraacinar pulmonary arteriolae (IAPA) in normoxic control group (0.1076 +/- 0.0205) was markedly weaker than that in chronic hypoxic group (0.3317 +/- 0.0683, t=3.125, P<0.05) and that in chronic hypoxic group was stronger than that in hypoxia plus L-arginine group (0.1928 +/- 0.0381, t=2.844, P<0.05). RT-PCR showed that the content of HIF-1alpha mRNA in chronic hypoxic group (2.5395 +/- 0.6449) was 2.16 times and 1.75 times higher than that in normoxic control group (1.1781 +/- 0.3628) and hypoxia plus L-arginine group (1.4511 +/- 0.3981), respectively. It is concluded that NO can reduce the mPAP by the inhibition of the expression of HIF-1alpha mRNA, which may be one of the mechanisms through which NO affects the pathogenesis of HPH.
Anoxia/metabolism ; Arginine/pharmacology ; Hypertension, Pulmonary/*metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; Nitric Oxide/*pharmacology ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Random Allocation ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors/*biosynthesis ; Transcription Factors/genetics

0.05). The production of TNF-? in LPS group was higher than that in control group (61 ng/L vs 156 ng/L, q=5.12, P