Construction of an adipose derived stem cell-based alphastatin peptide glioma targeting vector and its inhibitory effect on angiogenesis / 西安交通大学学报(医学版)

【Objective】 To construct an adipose derived stem cell-based alphastatin peptide glioma targeting vector and detect its anti-angiogenesis effect in vitro. 【Methods】 The adipose derived stem cell-based alphastatin peptide glioma targeting vector (Al-ADSCs) was constructed by transfecting the alphastatin peptide lentivirus vector into adipose derived stem cells (ADSCs). The expressions of stem cell markers on the surface of targeted vector were detected by flow cytometry. The expression of alphastatin peptide in the targeted vector and in the cell culture supernatant of the targeted vector were detected by Western blotting and ELISA, respectively. Cell migration assay was used to detect the tendency of the targeted vector toward CD133+ glioma stem cells, and lumen formation assay was used to detect the effect of the targeted vector on endothelial cell angiogenesis in vitro. 【Results】 After transfection, the surface markers of stem cells expressed by targeted vector did not significantly change compared with ordinary adipose derived stem cells. Western blotting showed that the targeted vector could successfully express alphastatin peptide. ELISA showed that the alphastatin peptide was detected in the cell culture supernatant of targeted vector \mg/L]. Cell migration test showed no significant difference in the tendency of CD133+ glioma stem cells between the targeted vector and ordinary adipose derived stem cells \. Lumen formation experiment showed that the targeted vector could inhibit endothelial cell-mediated angiogenesis in vitro [Lumen count: Control group (13.33±0.76)/HPF, ADSCs group (19.40±1.71)/HPF, Al-ADSCs group (7.27±0.31)/HPF, P<0.01]. 【Conclusion】 In the process of constructing the adipose derived stem cell-based alphastatin peptide glioma targeting vector, the stem cell biological characteristics and tumor tendency of targeted vector have no significant changes. This targeted vector can stably express and secrete alphastatin peptide and inhibit endothelial cell-mediated angiogenesis in vitro.

Surgical freedom of odontoidectomy via endoscopic endonasal approach / 西安交通大学学报(医学版)

【Objective】 To quantitatively analyze the surgical freedom of odontioectomy via endoscopic endonasal approach. 【Methods】 Seven adult head specimens were dissected by the endoscopic transnasal approach to the sellar region and craniocervical junction. The center of sellar floor (CenSF), opticocarotid recess (LOCR), foramen magnum, atlas, atlas-occipital joint and tip of odontoid process (TOP) were exposed. The surgical freedom of TOP was calculated by using the spatial coordinate positioning system of neuronavigation, and compared with that of LOCR and CenSF. 【Results】 CenSF and LOCR were common landmarks in the endonasal endoscopic approach. When the surgical freedom between TOP and CenSF and LOCR was compared, it indicated that ① The angle of attack on axial plane (AAAP):There was a significant difference among TOP, LOCR and CenSF (5.7 ° vs. 6.9 ° vs. 8.5 °, P=0.004). The comparison between the two groups showed that TOP was less than CenSF (P=0.003). ② The angle of attack on sagittal plane (AASP): There was a significant difference among TOP, LOCR and CenSF (6.3° vs. 7.0° vs. 9.5°, P=0.009). The TOP was less than CenSF (P=0.008). ③ There was no statistical significance between TOP and LOCR in surgical freedom (P=0.604, P=0.688). 【Conclusion】 Endoscopic transnasal approach can provide sufficient surgical freedom for odontoidectomy.

Effects of silencing AEG-1 gene by shRNA on vasculogenic mimicry of a glioma xenograft model / 西安交通大学学报(医学版)

【Objective】 To investigate the effects of silencing AEG-1 gene by shRNA on vasculogenic mimicry (VM) of a glioma xenograft model. 【Methods】 U87 glioma cells were infected with AEG-1 shRNA lentivirals. Real-time PCR and Western blotting were used to detect the mRNA and protein expressions of AEG-1 in U87 cells after infected by the AEG-1 shRNA lentivirals. A glioma xenograft model was generated and CD 34/PAS double-staining was performed to detect the VM channels in vivo. The immunohistochemical assay was performed to evaluate the expressions of MMP-2, MMP-9, VE-cadherin, and VEGF in glioma xenograft models. 【Results】 AEG-1 shRNA lentivirals could significantly inhibit the AEG-1 expression in glioma cells (P<0.01). Meanwhile, they also decreased the number of VM in the glioma xenograft model (P<0.01). Furthermore, the expressions of MMP-2, MMP-9, VE-cadherin, and VEGF in glioma significantly decreased in vivo (P<0.01). 【Conclusion】 These results suggest that silencing AEG-1 gene by shRNA can significantly inhibit VM of glioma in vivo, the mechanism of which may partly be through regulating MMP-2, MMP-9, VEGF, and VE-cadherin expressions.

Effect of madecassoside on hippocampal NR2B expression and cognitive function in rat traumatic brain injury model / 国际外科学杂志

Objective:To investigate the Effect of madecassoside (MC) on hippocampal NR2B expression and cognitive function following traumatic brain injury (TBI).Methods:One hundred and sixteen SD rats were provided by the experimental animal center of Medical College in Xi′an Jiao Tong University. Rats were randomly divided into four groups ( n=29) as following: Control group (Con group), TBI group, MC treated TBI rats group (TBI+ MC group) and MC treated control rats group (Con+ MC group). NR2B protein levels in hippocampus were detected by Western-blot at 12 h, 1 d, 3 d, 7 d, 14 d and 21 d post trauma. Hippocampal NR2B positive cells were studied by immunohistochemical (IHC) staining at 21 d post trauma. Cognitive functions of rats were evaluated by Morris water maze (MWM) test from 21 d to 25 d post trauma. Results:Expression of hippocampal NR2B in TBI group at 12 h, 1 d and 3 d post injury were 3.31±0.28, 2.17±0.31 and 1.96±0.31, which presented statistical difference among the three time-points ( P<0.05) and were significantly increased compared to Con group ( P<0.05). However, there was no difference of NR2B level between TBI group (0.93±0.13) and Con group ( P>0.05) at 7 d post injury. In addition, NR2B expression in TBI group at 14 d and 21 d post injury were 0.45±0.04 and 0.21±0.04, which were much lower than that in Con group. IHC staining revealed that the number of hippocampal NR2B positive cells in TBI group (33.26±9.71) were lesser than that of Con group (86.62±17.05). Increased Escape latency, decreased platform crossing times and target quadrant duration were found in TBI group compared with Con group ( P<0.05). Hippocampal NR2B expression in TBI+ MC group at 12 h, 1 d and 3 d post injury were 2.37±0.34, 1.38±0.22 and 1.14±0.16. Difference among the three time-points exhibited statistical significance ( P<0.05). In addition, they were much lower than that in TBI group at the same time-points ( P<0.05), but no difference was found between TBI+ MC group (0.97±0.06) and TBI group at 7 d post injury ( P>0.05). Moreover, NR2B expression in TBI+ MC group at 14 d and 21 d post injury were 0.86±0.08 and 0.52±0.06, which were much higher than that in TBI group ( P<0.05). IHC staining showed the number of hippocampal NR2B positive cells in TBI+ MC group (69.08±12.24) were much more than that of TBI group (33.26±9.71). A decrease of escape latency, an increase of platform crossing times and target quadrant duration were found in TBI+ MC group compared to TBI group ( P<0.05). Conclusion:MC treatment could inhibit the up-regulation of NR2B in hippocampus at early period of TBI and prevent the down-regulation of NR2B at advanced stage of TBI, which led to a remarkable improvement for the cognitive dysfunction caused by TBI.

Clinical study on craniopharyngioma resection via fronto-basal interhemisheric approach / 中国医师进修杂志

Objective To explore the operational methods and effect of craniopharyngioma resection via fronto-basal interhemisheric approach. Methods The clinical data of 31 patients who had underwent craniopharyngioma resection via fronto- basal interhemisheric approach were analyzed retrospectively. Results The saddle area structures of all the 31 patients were revealed well. The tumor total resection was in 19 cases (61.3%), and the tumor subtotal resection was in 12 cases (38.7%). The vision and field of vision after surgery were improved. Six cases (19.4% ) presented electrolyte disturbance, and 15 cases (48.4%) presented diabetes insipidus. Conclusions The fronto-basal interhemisheric approach can clearly expose the anatomical structures in midline and saddle area. This approach is safe and effective for craniopharyngioma resection.

Construction of lentivirus vectors carrying alphastatin gene and its secretion expression in human umbilical vein endothelia cells / 药物分析学报

Objective To construct lentivirus vectors carrying alphastatin gene, test its secretion expression in human umbilical vein endothelia cells (HUVECs) and observe its effects on growth, migration and tube formation of HUVECs. Methods We constructed recombinant lentivirus vectors of NT4-alphastatin fusion gene containing neurotrophin-4 signal peptide, pro-region sequences and alphastatin, then transfected the recombinant lentivirus vectors into HUVECs to obtain secretory protein alphastatin and test its anti-angiogenic activities in vitro. Results Our data showed that recombinant self-inactivating lentivirus vectors of NT4-alphastatin were successfully constructed, and stable NT4-alphastatin transduced HUVECs were capable of sustainably secreting alphastatin which significantly suppressed HUVECs migration and differentiation but not VEGF-induced proliferation. Conclusion This report represents the first time on the use of lentivirus-based vectors to deliver alphastatin, the endogenous angiogenesis inhibitor, and reveals the potential utility of anti-angiogenic gene therapy with lentivirus vectors for treating cancer.

Ethical thought on patients of head injury / 中国医学伦理学

The head injury patients is very common in clinic and it is complicated to cure them.Furthermore,many ethical questions are involved during the treatment,which are discussed in the article.The aim is to be helpful to protect both patients'and medical staff's right,reduce medical disputes and form a good relationship between doctors and patients from the view of ethics.Accordingly,the patients of head injury will acquire better treatment.