2.Construction and expression of wildtype and T34A mutant survivin retrovirus
Xiaoya YANG ; Jianxun SONG ; Yu BAI
Journal of Third Military Medical University 2003;0(08):-
Objective To construct the bicistronic retroviral vectors containing GFP gene and human survivin gene(wildtype and T34A mutant). Methods Survivin fragments were amplified from eukaryotic expression vectors by PCR, then cloned into pMIG retroviral vectors. The constructed recombinant was identified by double digestion with restriction enzymes. The constructed retroviral vectors was transfected into Phoenix E package cells under mediation of liposome. The expression of mRNA was examined by RT-PCR and the transcription of gene was determined by FACS analysis of GFP. Results The target fragments were successfully bound to pMIG vector. After the vectors were transfected into NIH3T3 cells, the expression of mRNA was confirmed by RT-PCR and the expression of GFP was confirmed by FACS analysis. Conclusion Human survivin gene wildtype and T34A mutant retroviral vectors have been constructed successfully, which can provide strong molecular tools for further studies and a novel approach for selective cancer gene therapy.
3.Analysis of remaining symptoms after discectomy with different operation for lumbar disc herniation
Yang YANG ; Weimin HUANG ; Ruoxian SONG ; Xiuchun YU
Chinese Journal of Postgraduates of Medicine 2015;38(7):486-490
Objective To analyze the incidence of remaining symptoms after discectomy with different operation,and investigate its causes and clinical significance.Methods Two hundred and twentyfour patients were followed up.The incidence of remaining symptoms and clinicai situation after discectomy were observed with different operation:discectomy (110 patients,single group),discectomy with bone grafting internal fixation (70 patients,fusion group),and discectomy with dynamic fixation (44 patients,non-fusion group).The SPSS 19.0 software was used to analyze the data.Results One hundred and sixpatients (47.32%,106/224) occurred remaining symptoms in 224 patients.In single group was 69 patients (62.73%,69/110),in fusion group was 26 patients (37.14%,26/70),and in non-fusion group was 11 patients (25.00%,11/44).The incidence of remaining symptoms among three groups was significant difference(x2 =22.177,P =0.000).The incidence of remaining symptoms in single group was significantly higher than that in non-fusion group and fusion group (x2 =17.921,P =0.000;x2 =11.235,P =0.001).The incidence of remaining symptoms in non-fusion group and fusion group had no significant difference (x2 =1.817,P =0.178).Conclusions There is significantly different in the incidence of lumbar disc herniation with different surgery methods.Discectomy with bone grafting internal fixation and with dynamic fixation can effectively reduce the incidence of remaining symptoms.
4.Clinical significance of CT perfusion image on post-operative follow-up of glioma
Fengchang YANG ; Aiqin SONG ; Haiying YU ; Wenwu LI ; Hongming SHEN
Journal of International Oncology 2009;36(2):155-158
Objective To assess the clinical value of multi-slice helical CT perfusion on the post-operative follow-up of glioma.Methods CT perfusion image was applied to 32 patients with glioma after operation.Various perfusion parameters,include cerebral blood volume(CBV),permeability surface(PS),relative cerebral blood volume(rCBV)and relative permeability surface(rPS)were measured on recurrent area,nonrecurrent area and normal cerebral area respectively.SPSS 12.0 statistical software was used.Independent t test was used to compare the differences.Results In the recurrent group(16 cases),the value of CBV in creased in 12 cases and was normal in the other 4 cases.The value of PS increased in all the cases of recurrent group.The value of rCBV exceeded 2.6 in 10 cases,among which the biggest value of rCBV was 4.01.The mean value of rCBV was 2.67 and the mean value of rPS was 6.32;In the non-recurrent group(16 cases),the value of CBV was nearly normal in 7 cases,increased in 2 cases and decreased in the remaining 7 cases.The value of PS was nearly normal in 11 case,decreased in 3 cases and increased in the remaining 2 cases.The values of rCBV were all less than 2.6 and the biggest value of rCBV was 2.14.The mean value of rCBV was 0.99 and the mean value of rPS was 1.42.The statistic significance was obtained for all the parameters when compared the recurrent group with the non-recurrent group(P<0.01)and with the normal cerebral area(P<0.01).When compared the non-recurrent group with the normal cerebral area,none of the parameters had statistic significance(P>0.05);The statistic significance was obtained for rCBV and rPS when compared the recurrent group with the non-recurrent group(P<0.01).Conclusion CT perfusion image is valuable in determining glioma recurrence after operation.
5.Three new xanthones from Garcinia xanthochymus
Fangfang ZHONG ; Yu CHEN ; Fajun SONG ; Guangzhong YANG
Acta Pharmaceutica Sinica 2008;43(9):938-941
To study xanthones from the barks of Garcinia xanthochymus, the constituents were isolated by normal-phase and reverse-phase silica gel column chromatography from the EtOAc extract. Their structures were elucidated by spectral analysis. Three new xanthones were purified and identified as 1,2,5-trihydroxy-6-methoxyxanthone (1), 1,4,6-trihydroxy-5-methoxyxanthone (2), 1,2,7-trihydroxy-4-( 1,1-dimethylallyl) xanthone (3).
6.Isolation and activity of an α-amylase inhibitor from white kidney beans
Xiaoqi ZHANG ; Mingyan YANG ; Yu MA ; Jia TIAN ; Jirong SONG
Acta Pharmaceutica Sinica 2007;42(12):1282-1287
An α-amylase inhibitor (α-AI) was isolated from white kidney beans (Phaseolus vulgaris. L) by ethanol fractional precipitation, ion exchange chromatography and gel filtration column chromatography. It was a homogeneity glycoprotein demonstrated by SDS-PAGE and gel filtration on CL-6B. The glycoprotein contained 88.2% protein and was rich in aspartic acid, glutamic acid, leucine, threonine and serine. The carbohydrate moiety was consisted of Man, Glc, Gal and Xyl in a mole ratio of 2.42∶1.50∶1.52∶1.00. The glycan and the core protein backbone was connected by O-linkage as determined by β-elimination reaction. The continuous oral administration of the α-AI (150 mg·kg-1·d-1 ) for 7 days can lower fasting blood glucose and 300 mg·kg-1 ·d-1 α-AI for 7 days can improve the sugar tolerance on alloxan-dependent diabetic model rats. The result showed the α-AI obtained from white kidney beans had good hypoglycemic effect on alloxan induced diabetic rats and may have high potential pharmaceutical value as a regulative digestive-starch degradation in patients suffering from diabetes.
7.The effect of 50 Hz sinusoidal electromagnetic fields on the proliferation of the osteosarcoma cell line MG-63
Lin SHANG ; Yang LIU ; Mingyu SONG ; Jizhe YU ; Hua WU
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(11):812-814
Objective To investigate the biological effect of 50 Hz sinusoidal electromagnetic fields at 1 mT on the proliferation of the human osteosarcoma cell line MG-63.Methods Osteosarcrma MG-63 cells were divided into control and experimental groups.The control group was incubated without an electromagnetic field; the experimental group was incubated in a 50 Hz,1.0 mT sinusoidal electromagnetic field.On the 2nd,4th and 6th day,their proliferation was determined using a cell counting kit-8(CCK-8)assay.Variations in the cell cycle were detected with flow cytometry(FCM).Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)was used to measure cyclin B1 and cyclin D1 mRNA.Results Compared with the control group,proliferation of the experimental group cells was reduced significantly.The percentage of cells at G0-G1 phase increased,and the mRNA expression of cyclin B1 and cyclin D1 was significantly reduced.Conclusions A 50 Hz sinusoidal electromagnetic field at 1.0 mT can inhibit the proliferation of osteosarcoma cell line MG-63 significantly.
8.Effect of different concentrations of uric acid on the neural differentiation of human bone marrow mesenchymal stem cells
Qingqing SONG ; Lingfan YU ; Lili XU ; Nailong YANG
Chinese Journal of Tissue Engineering Research 2014;(6):847-852
BACKGROUND:Uric acid as an endogenous antioxidant has garnered increasing attentions because of its anti-oxidation, anti-DNA damage and neuroprotective effects.
OBJECTIVE:To observe the effect of uric acid at different concentrations on the neural differentiation of bone marrow mesenchymal stem cells.
METHODS:Bone marrow mesenchymal stem cells were isolated, purified and cultured in vitro. The morphology change was observed. The third passages of bone marrow mesenchymal stem cells were induced to differentiate to neuron-like cells by induced liquid containing four different concentrations of uric acid (0 mmol/L as control group, 0.2 mmol/L, 0.4 mmol/L, 0.8 mmol/L) for 24 hours. Then, after second intervention for 1 hour, cells were detected by Nissl staining and specific markers were detected by immunohistochemistry method.
RESULTS AND CONCLUSION:After induction, the cellbody shrank, forming processes and connections. Nissl body was found in the cytoplasm. The positive rates of neuron-specific enolase were significantly higher in uric acid groups of different concentrations compared to the control group (P<0.05);moreover, the positive rates of neuron-specific enolase were increased as the increase in concentrations of uric acid (P<0.05). The positive rates of Nestin were decreased in uric acid groups of different concentrations compared to the control group (P<0.05). After 4 hours of induction, cells fel off significantly. In a certain period of time, uric acid can promote differentiation of bone marrow mesenchymal stem cells into neuron-like cells in a certain concentration-dependent manner in vitro.
9.A three-dimensional CT-based three-column classification of femoral intertrochanteric fractures
Xiaobing CHU ; Yu YANG ; Jianhua SONG ; Gaiping HAO ; Peijian TONG
Chinese Journal of Trauma 2013;29(11):1068-1073
Objective To introduce a new classification system for femoral intertrochanteric fractures and propose classification to assist surgical management.Methods A total of 176 cases of femoral intertrochanteric fractures treated from September 2007 to October 2011 were enrolled in the study.X-ray and three-dimensional spiral CT reconstruction were performed to define fracture shape,location,number of major fragments and displaced direction.As combined with the intraoperative findings and cadaveric anatomical study,the femoral intertrochateric fractures was simulated using a three-dimensional entity of normal proximal femur produced by Mimics 11.1 program.A new classification system for femoral intertrochateric fractures was proposed.Results The new system categorized intertrochateric fractures of the femur into five types (Type Ⅰ,Type Ⅱ,Type Ⅲ,Type Ⅳ and Type Ⅴ) based on the integrity or destruction of the medial column,lateral column and posterior column set and divided on the basis of bone connection of femoral head-neck fracture fragments with the bone in intertrochanteric region.Conclusions The new classification system with introduction of three-column concept is brief and practical and has a clear definition of fracture stability.Besides,the new classification-based treatment is recommended.
10.Effect of Yishen Jianpi Yangxue Tongli Fang on IGF-1 mRNA Expression of PCOS Rat
Yu-Hua HUANG ; Hong YANG ; Song-Yan CHAI ;
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(11):-
Objective To investigate the effects of Yishen dianpi Yangxue Tongli Fang (YSJP) on IGF-1 mRNA expression in the ovary of PCOS rat, and study its mechanism on the molecular level. Methods Seventy two female SD rats were randomly divided into normal control group, model group, clomifene group and low-, mid-, high-dose Chinese medicine group. The animal model established by capsulae levonorgestreli silasticus was used to observe the IGF-1 mRNA expression in ovary by hybridization in situ. Results IGF-1 mRNA expression of PCOS model group was higher than the normal control group obviously. The semi-quantitative image analysis showed that TCM treatment could decrease IGF-1 mRNA expression. Conclusion Yishen Jianpi Yangnue Tongli Fang could decrease the IGF-1 mRNA expression in ovary and improve the insulin resistance of PCOS rats.