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.A new device and method for tail vein injection in mice
Tao YU ; Yang ZENG ; Yuxia SUN ; Yongpeng SONG ; Xun TANG
Chinese Journal of Comparative Medicine 2016;26(10):82-84
Objective To provide a device and an effective method for tail vein injection in mice. Methods Doing the tail vein injection in mice with the self-designed device which is consisted of constant temperature part, lighting part and holding part.The difficulty and time of injection with and without the device were compared.Results It was faster and more accurate to perform the tail vein injection in mice with this self-designed device.Conclusion Using this self-designed device can significantly improve the efficiency and save the injection time.
5.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.
6.Effect of Chinese Medicinal Herbs on Human Scalp Hair Growth in Vitro
Yu SUN ; Shuxia YANG ; Ping TU ; Jiquan SONG ; Yangfeng DING
Chinese Journal of Dermatology 1994;0(05):-
Objective To study the effect of Chinese herbs on human scalp hair growth in vitro. Methods Organ culture of human scalp hair follicles was used to study the effects of extracts of Crataegus cuneata, Ligustrum lucidum, Polyporus umbelatus and Bletilla striata mixture and ginseng saponin on hair growth. Results Low doses of mixture extracts(1.28 ?g/mL and 6.4 ?g/mL) markedly enhanced the hair growth and lengthened the period of hair growth, while high doses of mixture extracts(4 mg/mL and 20 mg/mL) sharply inhibited hair growth and shortened the period of hair growth. High doses of ginseng saponin (40 ?g/mL and 200 ?g/mL) also had an inhibiting effect on hair growth. Conclusion This observation suggests that extracts of Crataegus cuneata, Ligustrum lucidum, Polyporus umbelatus and Bletilla striata mixture can promote hair growth in vitro.
7.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.
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.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 .
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.