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 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.
5.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 .
6.Morphea complicated by Hashimoto's thyroiditis in two sisters
Jinpeng LIU ; Xiaohong YU ; Guoling YANG ; Zhiqi SONG
Chinese Journal of Dermatology 2015;48(4):248-251
Morphea complicated by Hashimoto's thyroiditis is reported in two sisters.Case 1:a 64-year-old female presented with skin rashes on the anterior neck,trunk and bilateral anterior shins for 5 years,itching skin rashes on the perineum for 4 years,and Hashimoto's thyroiditis for 9 years.Physical examination revealed grade 1 enlargement of firm thyroid gland without exophthalmos or pretibial myxedema.Dermatological examination showed pink patches on the neck and breast,sclerosis and atrophy of skin over the back,porcelain-white patches on the perineum.Histopathological findings suggested the diagnosis of morphea on the breast and lichen sclerosus et atrophicus on the perineum.Case 2:a 55-year-old female,who was the younger sister of case 1,suffered from gradual sclerosis and atrophy of skin in the left inframammary region and abdominal region for 4 years,as well as Hashimoto's thyroiditis for 3 years.Similarly,physical examination revealed grade 1 enlargement of firm thyroid gland without exophthalmos or pretibial myxedema.Hypopigmentation,sclerosis and atrophy of skin were observed in the left inframammary region,abdominal region and central back region.Histopathological examination suggested a diagnosis of morphea.According to the clinical and histopathological manifestations,periodic acid-Schiff staining and thyroid gland function test results,the 2 cases were both diagnosed as morphea complicated by Hashimoto's thyroiditis.
7.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.
8.Effect of transketolase-like protein 1 on proliferation of human nasopharyngeal carcinoma cells
Song ZHANG ; Juhong YANG ; Weijia KONG ; Yu SUN
Chinese Journal of Pathophysiology 1986;0(03):-
0.05). However,the expression level of TKTL1 gene was significantly downregulated in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector. Cancer cells were arrested in G0/G1 phase,and cancer cell proliferation was significantly inhibited in the CNE cells transfected with siRNA TKTL1 construct. CONCLUSION:TKTL1 plays an important role in the total transketolase activity and cell proliferation of human nasopharyngeal carcinoma. TKTL1 may be considered as a potential target for novel anti-cancer therapy.
9.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.
10.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.