1.Experimental research of immunophenotype CD133,CD34,CD44 in human lung adenocarcinoma
Sichuang TAN ; Ruoxin WANG ; Sipin TAN ; Wen HU ; Yuchao MA ; Fenglei YU
Journal of Chinese Physician 2011;13(5):592-595
Objective To validate the possibility of CD133 CD34 CD44 be served as biomarkers in cancer stem cell of human lung adenocarcinoma. Methods Two kinds of culturing methods were performed to generate adhesive tumor cells and floating aggregates, and the differences of expression of CD133 CD34 CD44 between 2 kinds of cultured cells were observed by immunofluorescence. Results Floating aggregates grew more slowly, kept activity for longer period than adhesive cells (72.5% vs 47.5%,P<0.05). Floating aggregates expressed higher level of CD133, CD34 and CD44 than adhesive cells (68.97%,82.76%,93.10% vs 5.26%,15.79%,5.26%,P<0.01). Conclusions The combination of CD133, CD34 and CD44 probably can be used as surface markers of cancer stem cells for human lung adenocarcinoma.
2.Construction and evaluation of human Dp71 shRNA vector
Sichuang TAN ; Zhikang CHEN ; Xiaoxia CAO ; Qiaocheng WEN ; Weilin ZHANG ; Qingren ZENG ; Sipin TAN
Journal of Central South University(Medical Sciences) 2014;(4):338-343
Objective: To construct effective short hairpin RNA (shRNA) recombinant plasmids targeting humanDystrophin Dp71 gene, and evaluate their interference effciency. Methods: hTree pairs of siRNA sequences targeting human Dp71 gene and one pair of control siRNA sequence were designed, synthesized, and then inserted into the pRNAT-U6.1/Neo vector. hTe shRNA recombinant vectors were evaluated by enzyme digestion and sequencing. Dp71-shRNA and control shRNA plasmids were transfected into human normal gastric epithelial cells (GES-1) and humanbronchial epithelium (HBE). Western blot was used to evaluate its interfering effciency. Results: Restriction enzyme digestion and sequencing showed that the Dp71-shRNA vectors were successfully constructed. Western blot displayed that Dp71 protein expression was reduced to a signiifcant degree atfer transfection with the 3 Dp71-shRNA plasmids, and Dp71-shRNA2 plasmid inhibit the Dp71 expression most effciently. Conclusion: Dp71-shRNA vectors have been successfully constructed. The 3 Dp71-shRNA plasmids can inhibit Dp71 expression in GES-1 and HBEC, with Dp71-shRNA2 plasmid displaying the highest inhibition effciency.
3.Effect of nucleolin on cardiac cell apoptosis in Type 2 diabetic cardiomyopathy mice
Meidong LIU ; Li SUN ; Bimei JIANG ; Sipin TAN ; Ke LIU ; Xianzhong XIAO
Journal of Central South University(Medical Sciences) 2017;42(3):241-245
Objective:To investigate the effect of nucleolin on cardiac cell apoptosis in Type 2 diabetic cardiomyopathy mice.Methods:Mice were fed with high-fat and high-sugar food for 20 weeks (mice were injected intraperitoneally with 60 mg/kg streptozotocin in the 5th and 6th weeks) to establish a mouse model of Type 2 diabetes.The mice were divided into 4 groups:a wild type (WT) control group,a nucleolin transgenic (TG) control group,a WT diabetic group,a TG diabetic group.Diabetesrelated indicators were detected at the end of the 8th week.At the end of the 20th week,HE staining was used to observe myocardial morphological changes;TUNEL staining and caspase-3 activity were used to detect the extent of apoptosis of cardiac myocytes.Results:The level of fasting blood glucose was significantly increased in the diabetic group than that in the control group.In WT diabetic group,myocardial disarrangement,fragmentation and dissolution were observed (determined by HE staining);cellular apoptosis (determined by TUNEL staining and caspase-3 activity) also increased markedly in the WT diabetic group.Compared with the wild mice in the diabetic group,myocardial morphological changes and cardiac myocytes apoptosis were alleviated significantly.Conclusion:Nucleolin overexpression affectes the occurrence and development of diabetic cardiomyopathy through inhibition of cardiac myocyte apoptosis.
4.LUNX mRNA in regional lymph nodes of non-small cell lung cancer patients by RT-PCR and its clinical significance.
Sichuang TAN ; Zhangbo CHENG ; Yuchao MA ; Sipin TAN ; Zhaochu YIN ; Wen HU ; Fenglei YU
Journal of Central South University(Medical Sciences) 2010;35(12):1236-1241
OBJECTIVE:
To evaluate the detection of humen-lung-specific X protein (LUNX) gene in micrometastases of patients with non-small cell lung cancer.
METHODS:
The expression of LUNX gene in tumor tissue, lung and lymph nodes was detected by reverse transcriptase-polymerase chain reaction(RT-PCR) both in 43 non-small-cell lung cancer patients (the experimental group) and 15 lung benign patients (the control group). LUNX mRNA expression in clinic pathology,stage of cancer cell differentiation, clinic stage, age, sex, smoking history, and 4 lung cancer blood markers (CEA,CA125,NSE, and CYFRA211) were evaluated.
RESULTS:
The expression of LUNX gene was positive in the 2 groups. LUNX gene expression was positive in 33 of the 87 lymph nodes of the 43 patients in the experimental group (37.93%), and in 2 of the 26 lymph nodes in the control group (7.69%). The LUNX mRNA positive in the lymph nodes was closely related to the pathological type, cancer cell differentiation and clinic stage(r=0.660,0.500,0.460; P=0.011,0.017,0.022, all P<0.05), while not closely related to age, sex, smoking history and 4 lung cancer blood markers (CEA,CA125, NSE, and CYFRA211) (r=0.111, 0.135,0.083,0.354; P=0.739,0.714,0.773,0.125,all P>0.05).
CONCLUSION
The LUNX mRNA expression detected by RT-PCR is more sensitive than by traditional ways. The expression of LUNX gene mRNA in the lymph nodes is a valuable index for the detection of micrometastases in patients with non-small cell lung cancer.
Adult
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Aged
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Biomarkers, Tumor
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biosynthesis
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genetics
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Carcinoma, Non-Small-Cell Lung
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genetics
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metabolism
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pathology
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Female
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Glycoproteins
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genetics
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metabolism
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Humans
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Lung Neoplasms
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genetics
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metabolism
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pathology
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Lymph Nodes
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metabolism
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Lymphatic Metastasis
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Male
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Middle Aged
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Phosphoproteins
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genetics
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metabolism
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RNA, Messenger
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genetics
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metabolism
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Reverse Transcriptase Polymerase Chain Reaction
5. Evaluation on case/problem-based and interactive teaching mode in pathophysiology curriculum
Sipin TAN ; Zihui XIAO ; Zizhi TU ; Kangkai WANG ; Ying LIU ; Bimei JIANG ; Gonghua DENG ; Huali ZHANG ; Xianzhong XIAO
Chinese Journal of Medical Education Research 2019;18(9):884-888
Objectives:
To provide new ideas on how to shift students' learning attitude from passive learning to active learning, we explored and evaluated a case/problem-based and interactive teaching mode in pathophysiology curriculum.
Methods:
Case/problem-based and interactive teaching mode is an innovative teaching model adopted in pathophysiology curriculum for grade 2015 students of 5-year program in clinical medicine and other medical students of non-clinical majors in Xiangya Medical School, Central South University. The teaching effectiveness of the case/problem-based and interactive teaching mode was evaluated by questionnaire survey, with 460 medical students enrolled in the survey whose approval degree on current teaching mode was analyzed. Excel was used to collect and process data, complete descriptive analysis and calculation of the percentage of indicators.
Results:
A total of 460 anonymous questionnaires were distributed and 453 valid questionnaires were retrieved, from which the following information was obtained: ① Pre-class learners' guidance designed for current teaching mode: 88.7% of students (402/453) believed that "Pre-class Learners' Guidance" motivated them to preview relevant teaching contents before class. 82.8% of students (375/453) believed "Pre-class Learners' Guidance" improved discussion quality in class. 76.6% of students (347/453) believed "Pre-class Learners' Guidance" expanded thinking and exploring space, while it did not increase student study burden (306/453, 67.6%). ② Compared with traditional teaching mode, the case/problem-based and interactive teaching mode had following advantages: It's helpful to cultivate students' clinical thinking (414/453, 91.4%), strengthen students' memory and understanding during study (400/453, 88.3%), attract students' attention in class (380/453, 83.9%), and aroused student's interest in class discussion (327/453, 72.2%). ③ 83.4% of students (379/453) preferred current teaching mode: they believed this teaching mode could improve students' ability to analyze and solve problems (325/453,71.7%), train clinical thinking (321/453, 70.9%), improve students' self-study ability (247/453, 54.5%) and increase students' capabilities of making summary and conclusion (197/453, 43.5%).
Conclusion
Case/problem-based and interactive teaching mode in pathophysiology curriculum enhances students' ability of self-studying, activates classroom's atmosphere, improves teaching quality, and effectively fosters students' clinical thinking. Therefore, this teaching mode deserves to be spread and applied in classroom teaching of pathophysiology and other basic medicine disciplines as well.