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.The value of clinical application of leukocyte classification alarm system (Q-Flag) for Sysmex XT-1800i blood cell analyzer
Bing HAN ; Yang GAO ; Kang LI ; Qisheng SONG ; Bin YU
International Journal of Laboratory Medicine 2015;(14):2013-2014
Objective To study the value of clinical application of leukocyte classification alarm system (Q‐Flag ) for Sysmex XT‐1800i blood cell analyzer .Methods 394 blood samples with alarm system (Q‐Flag) and 190 ones without abnormal alarm infor‐mation detected by the Sysmex XT‐1800i blood cell analyzer were performed to observe the quantity and morphology of blood cells by manual differentiation ,and the results were compared between them .Results The alarm system of Sysmex XT‐1800i blood cell analyzer in detecting cell morphologic abnormalities was 100 .0% for sensitivity ,62 .3% for specificity ,70 .8% for positive predic‐tive value and 100 .0% for negative predictive value .Based on thegold standardof manual differentiation ,no abnormal cells were observed in those blood samples without Q‐Flag alarm information and the coincidence rate was 100% .The coincidence rate of leu‐kocyte classification was from 0 .0% to 75 .0% for blood cell analyzer when the alarm system (Q‐Flag) was between 100U and 200U ,and that was from 66 .7% to 95 .6% when the alarm system (Q‐Flag) was between 200U and 300U .Conclusion The alarm system sensitivity of leukocyte classification alarm system (Q‐Flag) is high for Sysmex XT‐1800i blood cell analyzer ,and it is nec‐essary to manually differentiate the samples with abnormal Q‐Flag in order to provide accurate and reliable clinical diagnostic infor‐mation .
5.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.
6.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.
8.The expression of ZNRF3 in thyroid cancer and its functions in thyroid cells
Wangwang QIU ; Zhili YANG ; Jun YAN ; Song YU ; Qi ZHENG
Journal of International Oncology 2016;43(4):246-249
Objective To detect the expression and function of ZNRF3 in different kinds of thyroid cancer tissues and cells.Methods Immunohistochemistry was used to detect the expressions of ZNRF3 protein in 35 cases of papillary thyroid carcinoma and 10 cases of poorly differentiated thyroid carcinoma.The expressions of ZNRF3 gene in TPC-1 and 8505C were detected by RT-PCR,and the cell lines were derived from papillary thyroid carcinoma and poorly differentiated thyroid carcinoma respectively.After silenced ZNRF3 gene expression with lentivirus,the proliferation ability of TPC-1 cells were detected with CCK-8,the invasion and metastasis ability of TPC-1 cells were detected with Transwell.Results According to results of immunohistochemistry and RT-PCR,the expressions of ZNRF3 in papillary thyroid carcinoma cells and tissues were higher than those in poorly differentiated thyroid carcinoma cells and tissues,the differences were statistically significant (4.83±0.44 vs.3.13 ±0.59,t =2.20,P <0.05;1.01±0.06 vs.0.21±0.04,t =11.80,P<0.01).After ZNRF3 geng silencing,according to the results of CCK8,the proliferation ability of TPC-1 cells was significantly enhanced in 72 h,the difference was statistically significant (0.96 ± 0.10 vs.0.64 ± 0.05,t =3.19,P < 0.05);and according to the results of Transwell,the TPC-1 cell's invasion (0.12 ± 0.01 vs.0.09 ±0.00,t =5.48,P<0.01) and migration (0.22 ±-0.01 vs.0.17 ±0.01,t =4.58,P <0.05) also increased,the differences were statistically significant.Conclusion The expression of ZNRF3 in papillary thyroid carcinoma is higher than that in poorly differentiated thyroid cancer.ZNRF3 is tumor suppressor gene in the thyroid tumors.
9.The Differential Expression of MMP-9 and Ki67 in Pancreatic Cancer
Yu SUN ; Ning SUN ; Shaoshi YANG ; Guodong SONG ; Jianqiu CHEN
Tianjin Medical Journal 2014;(1):44-46
Objective To investigate the expression and significance of MMP-9 and Ki67 for predicting the progres-sion and prognosis of pancreatic cancer. Methods S-P immunohistochemical method was used to detect the expressions of MMP-9 and Ki67 in 100 pancreatic cancer tissue specimens. The relationship between the expressions of MMP-9 and Ki67 and patient age, tumor size, lymph node metastasis, tumor differentiation, clinical stages and prognosis were analyzed. Re-sults There were higher expressions of MMP-9 protein 46%(46/100) and Ki67 protein 53%(53/100) in 100 samples of pancreatic adenocarcinoma. And the expressions of MMP-9 and Ki67 were inversely associated with tumor differentiation, clinical stages, and lymph node metastasis of pancreatic cancer (P<0.05). There were no significant differences in the ex-pressions of MMP-9 and Ki67 between patient age and tumor size. The expressions of MMP-9 and Ki67 were positive corre-lated (rs=0.405,P<0.05). Moreover, the overall survival rates were correlated with patient age, lymph node metastasis, tumor differentiation and the expression of MMP-9, but no correlation with tumor size, clinical stages, and the expression of Ki67. Conclusion The expressions of MMP-9 and Ki67 were associated with pancreatic cancer progression. And the detection of expression of MMP-9 may have practical value in prognosis in patients with pancreatic cancer.
10.Effect of electroacupuncture at Neiguan (PC 6) and Baihui (GV 20) on CHOP and caspase-12 gene expressions in rats after ischemia-reperfusion injury
Wenhui LI ; Jie YU ; Yaping LIN ; Xing TAN ; Yang SONG
Journal of Acupuncture and Tuina Science 2017;15(1):8-13
Objective:To investigate the effects of electroacupuncture (EA) at Neiguan (PC 6) and Baihui (GV 20) by observing the changes of CCAAT/enhancer-binding protein(C/EBP) homologous protein (CHOP) and caspase-12 gene expressions in rats after cerebral ischemia-reperfusion injury (IRI), and explore whether the apoptosis pathway of endoplasmic reticulum stress (ERS) is involved in the protective mechanisms of EA.
Methods:Sixty rats were randomly assigned to five groups (12 in each group): a normal control group (group A), a sham-operation group (group B), an operation group (group C), an Edaravone group (group D) and an EA group (group E). The cerebral IRI rat model was induced by middle cerebral artery occlusion (MCAO) using intraluminal monofilament. 2,3,5-triphenyl tetrazolium chloride (TTC) staining was adopted in the measurement of cerebral infarction volume. Real-time polymerase chain reaction (RT-PCR) was used to determine the mRNA expressions of CHOP and caspase-12.
Results: Compared with group A and group B, the volume of cerebral infarction and mRNA expressions of CHOP and caspase-12 in group C, group D and group E were increased, with statistical significances (P<0.05 orP<0.01); compared with group C, the volume of cerebral infarction and mRNA expressions of CHOP and caspase-12 in group D and group E were decreased significantly (P<0.05 orP<0.01); there were no significant differences between group D and group E in comparing the above items (P>0.05).
Conclusion:EA at Neiguan (PC 6) and Baihui (GV 20) can effectively suppress the volume of cerebral infarction. Furthermore, the underlying mechanism of EA at Neiguan (PC 6) and Baihui (GV 20) is possibly related to the down-regulation of CHOP and caspase-12 mRNA expressions, so as to decrease cell apoptosis.