1.Color Doppler imaging feature of localized Castleman disease
Luzeng CHEN ; Liangliang ZHANG ; Bin WANG ; Hui ZHANG ; Yan XIONG
Chinese Journal of Ultrasonography 2012;21(8):701-703
Objective To summary the color Doppler imaging feature of localized Castleman disease.Methods From January 1997 to November 2011,32 localized Castleman diseases which were proved by pathology were analyzed.Results Round-like,hypoechoic,hypervascular lesions were showed in 23 hyalinevascular type lesions,2 of them with calcium,3 of them with structure liked lymph node hilum.Round-like,hypoechoic,hypervascular lesions or normal lymph node were showed in 5 plasma- type lesions and 4 mixed type lesions.Conclusions When round like,hypoechoic,hypervascular lesion is found by ultrasonography,Castleman should be considered.Calcium or lymph nod hilum-like structure is special finding in diagnosis of Castleman disease by ultrasonography.
2.Preparation of a G418-resistant feeder layer of NIH3T3 cells
Meiying ZHANG ; Hua LI ; Wanwei DONG ; Wei YANG ; Ying WANG ; Ying QIN ; Luzeng WANG ; Taiyi WANG
Chinese Journal of Tissue Engineering Research 2006;10(37):144-147,封三
BACKGROUND: Establishment of drug (neomycin or hygromycin) resistant feeder layer cells is necessary for screening the target gene positive clone of embryonic stem cell (ESC) transfected in vitro, and the establishment of drug-resistant NIH3T3 cell line is also important for the screening of other target ESC genes.OBJECTIVE: To obtain a feeder layer of positive cell clone of ESC transfected by pTet-on gene by means of the G418-resistant NIH3T3 cell line establishment.DESIGN: Cell culture and DNA examination.SETTING: Faculty of Laboratory Animal, China Medical University.MATERIALS: NIH3T3 cells were contributed by the Cell Biology Staff Room of China Medical University, and pWL/neo plasmid was a gift from Professor Jin Zhuang of Harvard University Medical College. G418,leukemia inhibitory factor (LIF, ESC GRO 106 U/mL) and DMEM were produced by GIBCO BRL Company, while mitomycin-C (MIT-C), DIG marking and kits were the products of Roche Company. Lipoetin was the product of Invitrogen Company, and bought from Shenyang Lianxing Biotechnology Co.,Ltd.METHODS: ①The pWL/neo plasmid containing neo gene was purified and transfected into NIH3T3 cells by lipoetin.②The transfected cells were further cultured in 25 mL medium containing G418 antibiotic of gradient concentration to observe the survival and apoptosis of cells and measure G418 minimal fatal dose (MFD) to NIH3T3 cells.③The transfected cells were subcultured, and the clone of single cell was selected to 24-pore culture board for screening amplification. At the same time, normal NIH3T3 cells were also selected and added with the selective culture medium at the same dose of G418, as screening negative control. ④The cells were planked with lower cell density, and further screened in DMEM medium containing G418 MFD; Meanwhile, normal NIH3T3 cells were taken as controls, and cultured by G418 contain MFD and by DMEM medium without G418 respectively. Light microscope was used to observe G418R NIH3T3 cells. ⑤G418R NIH3T3 cells and MEF were all applied as the feeder layer to culture the ES-D3 cells, which were observed by light microscope. Cell DNA was prepared, and evaluated by PCR and southern blot.MAIN OUTCOME MEASURES: ①G418 MFD to NIH3T3 cells. ②screening result of G418R NIH3T3 cells. ③growth of G418R NIH3T3 cells. ④growth of ES-D3 cells on G418R NIH3T3 cells.RESULTS: ①G418 MFD to NIH3T3 cells was defined as 500 mg/L.②Under the condition of G418 (500 mg/L), G418R NIH3T3 cell clones were successfully selected. ③The G418R NIH3T3 cells had no difference from normal NIH3T3 cells in morphology and propagation rate.④The ESC cultured in feeder layer present a colony growth, smooth limitation and remained an undifferentiation state. The resistant cell genomic DNA of G418R NIH3T3 cells was assayed with specific neo gene primer, and then neo gene DNA fragment was amplified; Southern blot analysis showed that neo gene fragment integrated into G418R NIH3T3 cells.CONCLUSION: The G418R NIH3T3 cells are established successfully,and the ESC cultured in the G418R NIH3T3 cell feeder layer can keep the characteristics of normal ESC.
3.Construction of a 5-Lipoxygenase Transgenic Mice
Meiying ZHANG ; Honglian WU ; Wei YANG ; Zhaoyang LI ; Wanwei DONG ; Shenglai ZHOU ; Yang YU ; Wei WANG ; Xiangchuan Lü ; Ying QIN ; Zhihong ZHENG ; Luzeng WANG
Acta Laboratorium Animalis Scientia Sinica 2010;18(1):60-64
Objective To construct a 5-lipoxygenase (5-LO) transgenic mouse model of atherosclerosis.Methods Purified 5-LO fragment was injected into male pronucli and the firtilized eggs were transplanted into pseudopregnant mice.PCR and Southern blot were used to detect the genotype of DNA separated from the newborn mouse tail tissues.RT-PCR and Western blot analysis were used to detect the gene transcription and expression.Results PCR and Southern blot results showed that 7 of 25 mice were transgenic mice.Expression of 5-LO and FLAP was found in the bone marrow,spleen,kidney,and peritoneal cells.Results of RT-PCR and Western blot showed that No.9,20,24transgenic mice expressed a higher level of 5-LO and FLAP than those in the wild type C57BL/6 mice.The expression levels in bone marrow and peritoneal cells were significantly different(P<0.05).Conclusion A 5-LO transgenie mouse line has been established in this study and may be used for future study on the function of 5-LO gene.
4.Dynamic study of platelets surface glycoprotein in Kawasaki disease.
Yi PANG ; Hetao LIANG ; Shilu YU ; Xiaoping LIU ; Quan WANG ; Fang TANG ; Yuhong LIU ; Luzeng JIANG
Chinese Journal of Hematology 2002;23(3):134-137
OBJECTIVETo study the activation of platelets in children with Kawasaki disease (KD) at molecular level.
METHODSThe expression of platelet surface glycoproteins CD(41), CD(42a), CD(61), CD(62p) and CD(63) in 20 KD patients was measured by flow cytometry before and at 1, 2, 3 week after treatment with aspirin and high-dose (1 approximately 2 g/kg) intravenous gamma-globulin (IVIG).
RESULTSThe expression of glycoprotein CD(41), CD(42a), CD(61), CD(62p) and CD(63) were higher in KD group than in control group. Aspirin and IVIG could not inhibit these high expression of glycoproteins. Higher expression of CD(62p) was observed in patients with coronary artery injury.
CONCLUSIONPlatelets were highly activated in KD patients which may be one of the most important pathophysiological step in KD. It provided a theoretical basis for treatment of KD with antagonist of glycoprotein of platelets. Obviously increase of CD(62p) can be taken as a criterion for predicting coronary artery injury in KD patients.
Aspirin ; pharmacology ; Blood Platelets ; drug effects ; metabolism ; Child ; Child, Preschool ; Coronary Artery Disease ; complications ; metabolism ; Female ; Humans ; Injections, Intravenous ; Male ; Mucocutaneous Lymph Node Syndrome ; metabolism ; pathology ; P-Selectin ; metabolism ; Platelet Activation ; physiology ; Platelet Membrane Glycoproteins ; metabolism ; gamma-Globulins ; pharmacology
5.Analysis of detecting value of ultrasound and the clinic-pathological features of axillary metastasis in breast cancer.
Ling XIN ; Luzeng CHEN ; Hong ZHANG ; Qian LIU ; Ling XU ; Bin WANG ; Ting LI ; Xuening DUAN ; Yinhua LIU
Chinese Journal of Surgery 2014;52(12):924-928
OBJECTIVESTo evaluate the value of ultrasound (US) in predicting axilla status and to investigate the clinic pathologic characters in the axillary node metastasis.
METHODSFrom June 2012 to June 2013, 323 female primary breast cancer patients who received both axilla ultrasound and pathology examinations were reviewed retrospectively. The features of axillary nodes including diameter, longitudinal-transverse axis ratio, cortical thickness and blood flow grade were used to evaluate axillary status. US accuracy of axillary node metastasis was analyzed correlated with the final pathology results. The clinical and histological features associated with axillary node metastasis was analyzed by χ² test.
RESULTSThe proportions of Luminal A-like, Luminal B-like, human epidermalgrowth factor receptor-2 positive and triple negative breast cancer were 11.1% (36/323), 58.5% (189/323), 13.3% (43/323) and 17.0% (55/323) . The sensitivity, specificity, positive predictive value and negative predictive value of axilla US in the diagnosis of nodal metastasis were 35.6% (46/129), 98.9% (181/183), 95.8% (46/48) and 68.6% (181/264). Axillary lymph node metastasis had statistically significant correlation with menopausal status and clinical tumor size (χ² = 4.337, 11.100; P = 0.037, 0.001).
CONCLUSIONSStandardized ultrasound is the basic way to evaluate axilla status. Sentinel lymph node biopsy should be done to acquire accurate preoperative staging of axilla when US shows no signs of metastasis. Axillary lymph node metastasis is significantly related to menopausal status and clinical tumor size, but not significantly related to subtype classification of primary breast cancer.
Axilla ; Breast Neoplasms ; pathology ; Female ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; diagnostic imaging ; Neoplasm Staging ; Retrospective Studies ; Sensitivity and Specificity ; Sentinel Lymph Node Biopsy ; Ultrasonography