2.Determination of Epicatechin Content in Uncaria macrophylla by HPLC
Jun YANG ; Chunqing SONG ; Ming CHEN ; Haiyan WANG ; Zhibi HU
Chinese Traditional and Herbal Drugs 2000;31(9):662-663
Epicatechin content in Uncaria macrophylla Wall. was determined by RP-HPLC. As aresult, the epicatechin contents were 0.38% in the hook and stem, and 0. 820% in the leaf. Thus it seemedto be more worthwhile to produce epicatechin from the leaf rather than from the hook and stem.
3.An Improved Method of Fluorescent Labeled Amplified Fragment Length Ploymorphism
Kai YANG ; Wan SONG ; Chunqing ZHANG ; Jizeng JIA
Progress in Biochemistry and Biophysics 2001;28(2):256-258
A new and efficient reaction system has been set up, in which the MseⅠ primers were fluorescent labeled for auto-sequencer. PCR reagents and primers and adapters of MseⅠ and EcoRⅠ, which were synthesized, the AFLP protocol has been modified, and reaction and electrophoresis conditions were optimized, the results obtained can be comparable to that of AFLP fluorescent labeled AFLP kits with less cost.
4.Effect of Low Frequency Ultrasound on Carotid Plaque
Fengyun WANG ; Yanhui LI ; Xiafeng YANG ; Chunqing FENG ; Yihua SUN ; Xiaoqian YANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(5):591-594
Objective To observe the effects of low frequency ultrasound on carotid artery plaque and artery stenosis. Methods 156 patients with carotid atherosclerosis were divided into treatment group (n=80) and control group (n=76). The control group was administered routine medicine, while the treatment group accepted low frequency ultrasound therapy in addition. The size and shape of carotid artery plaque, severity of stenosis and the level of lipid were observed before and after treatment, and the side-effects were recorded. Results The intima-media thickness (IMT), diameter of plaque, plaque score decreased after treatment in both groups, and decreased more in the treatment group than in the control group (P<0.05); while the frequence of moderate stenosis and severe stenosis was less (P<0.05). The levels of low density lipoprotein- cholesterol and total cholesterol decreased in both groups after treatment (P<0.05), and decreased more in the treatment group than in the control group (P<0.05). No serious side-effect was observed. Conclusion Low frequency ultrasound can reduce the atherosclerotic plaques in carotid artery and relieve the stenosis.
5.Effects of Bmi-1-siRNA on proliferation of lung adenocarcinoma SPC-A1 cells and its mechanism
Yifang WANG ; Ben LIU ; Chunqing LIU ; Xiangyu ZHENG ; Dandan LIU ; Jie ZHU ; Chunhui YANG ; Xiuxiang MENG
China Oncology 2014;(5):333-341
Background and purpose:The human oncogene B-cell-speciifc moloney murine leukemia virus integration site 1 (Bmi-1) is an important member of the polycomb group family, and it regulates cell proliferation and senescence via INK4a/ARF locus. This study investigated the effects of Bmi-1-siRNA on the proliferation of lung adenocarcinoma cell line SPC-A1 cells with INK4a/ARF locus and clarify the mechanism of Bmi-1-mediated effect on proliferation of lung adenocarcinoma cells. Methods:In this study, we chose the most efifcient siRNA chain the pGeneshl-2-Bmi-1 sense-1 and inserted into a pSUPER-retro-neo retroviral vector. The packaged si-Bmi-1 pSUPERret-ro-neo retroviral vector was stably transfected into lung adenocarcinoma SPC-A1 cell line. The stably transfected cells were cultured and passed. After transfection, the levels of Bmi-1 mRNA and protein expression of SPC-A1 cells were analyzed by RT-PCR and Western blot respectively. Trypan blue, MTT and plate colony forming assay were performed to observe the proliferation capibility of SPC-A1 cells and evaluate the cloning forming ability in vitro. The potency of tumorigenesis was observed in nude mouse through hypodermic inoculation of SPC-A1 cells. Cell cycle distribu-tion was analyzed by lfow cytometry (FCM) in SPC-A1 cells. The expression levels of proliferation proteins including p16INK4a, p53, Cyclin D1, PTEN, Akt and Ser473p-Akt were analyzed by Western blot. Results:The mRNA and protein expression levels of Bmi-1 were signiifcantly reduced in SPC-A1-Bmi-1-siRNA cells transfected with pSUPER-retro-neo retroviral vector. Knockdown of Bmi-1 could inhibit the growth, colony formation in vitro and tumorigenesis in vivo of SPC-A1 cells (P<0.01). The transfected SPC-A1 cells were arrested in G1 phase [(64.6±1.2)%, P<0.05]. Compared with two control groups, p16INK4a, p53 and Akt were not affected (P>0.05), while Cyclin D1 and Ser473p-Akt were downregulated (P<0.01) and PTEN was up-regulated (P<0.01) in the SPC-A1-Bmi-1-siRNA cells. SPC-A1-Bmi-1-siRNA cells were treated with various concentrations of PTEN inhibitor to determine expression levels of PTEN, Bmi-1 and Ser473p-Akt protein. Ablation of PTEN rescued Bmi-1 and Ser473p-Akt expression in SPC-A1-Bmi-1-siRNA cells. Conclusion:Knockdown of Bmi-1 gene can arrest the proliferation of SPC-A1 cells through G0/G1 phase arrest by inhibiting Cyclin D1 expression indirectly, which may be not associated with p16INK4a signaling pathway.
6.Seeding fresh bone marrow aspirate directly on scaffolds to construct tissue-engineered ligament
Hong WANG ; Ming TANG ; Chunqing MENG ; Hanqi WANG ; Shuhua YANG ; Jingyuan DU ; Deyu DUAN ; Zengwu SHAO
Chinese Journal of Tissue Engineering Research 2014;(12):1870-1876
BACKGROUND:There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned.
OBJECTIVE:To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament
METHODS:We constructed fibroin fiber/smal intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cel s (bone marrow mesenchymal stem cel s) on the scaffold to built cel seeding group. The control group had no treatment. After that, we detected the density of cel adhesion, cel proliferation ability and extracel ular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks.
RESULTS AND CONCLUSION:After 4 hours of incubation, bone marrow seeding group was significantly higher than the cel seeding group in cel adhesion density and proliferation rate (P<0.05). Bone marrow seeding group and cel seeding group showed higher type I, III col agen secretion compared with the control group (P<0.05), but the col agen secretion of bone marrow seeding group and cel seeding group showed no significant difference. Composite cel scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.
7.Blood oxygen level and the pathogenic mechanism of expressive aphasia after stroke
Huixiang WU ; Weihong QIU ; Zhuang KANG ; Chunqing XIE ; Guifang WAN ; Qinglu YANG ; Shaoqiong CHEN
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(6):407-412
Objective To evaluate the activation patterns in the cortexes of expressive aphasics after stroke so as to explore the pathogenic mechanism of expressive aphasia.Methods Blood oxygen level dependent functional magnetic resonance imaging (BOLD-fMRI) was the method of choice.It was administered to 9 subjects with expressive aphasia at 1 to 3 months post-stroke onset and to 10 healthy controls.Active areas in the patients' brains were observed using a block-designed picture-naming task,and language function was tested with the China Rehabilitation Research Center's aphasia examination (CRRCAE).The control group received BOLD-fMRI only.SPM8 software was used to process the fMRI data.Results Differences were observed in the mapping of activated areas between the two groups,but many activated areas showed no difference.Significant differences in activation were observed in areas associated with vision,language and cognition,including the bilateral inferior frontal gyrus,the bilateral superior temporal gyrus,the bilateral insula,the bilateral basal ganglia,the left superior frontal gyrus,the left middle frontal gyrus,the left precentral gyrus,the left thalamus,and the left middle temporal gyrus.All the patients had activated cortex regions associated with visual processing in the left and/or right hemisphere,such as the middle frontal gyrus,the middle temporal gyrus,the lingual gyrus and the fusiform gyrus.The activation volumes in the left hemisphere were significantly smaller than those in normal adults.Regions related to language such as the left inferior frontal gyrus (Broca's area),the left middle frontal gyrus,and the right inferior frontal gyrus (the mirror region of Broca's area) were activated in some of them.While the activation frequency,activated volume and activation intensity generally were all less in the patients than in the controls,the activation intensity in the right superior temporal gyrus,the bilateral superior parietal lobule and the left inferior temporal gyrus were stronger.Conclusions Language production may be associated with multiple,interconnected regions.The right hemisphere participates in natural language processing.Aphasia damages both linguistic and cognitive areas,reducing activation in Broca's aphasia.Activation areas in the left hemisphere and the right inferior frontal gyrus decrease significantly,while some regions in the right hemisphere are relatively more activated.The right inferior frontal gyrus may play a different role in language recovery at different periods of aphasia after stroke.
8.Effect of siRNA-mediated silencing Bmi-1 gene expression on the proliferation of lung cancer cell line A549 in vitro and in vivo
Xiangyu ZHENG ; Jie ZHU ; Yifang WANG ; Chunqing LIU ; Ben LIU ; Chunhui YANG ; Dandan LIU ; Xiuxiang MENG
China Oncology 2013;(7):505-511
Background and purpose:The pro-oncogene Bmi-1 is a member of the polycomb-group family, can regulation of the proliferation and self-renewal of normal and tumor stem cells. In recent years, Bmi-1 has been found that it is overexpressed in varieties of human malignant tumors. The study aimed to observe the effects of Bmi-1-siRNA on the growth capacity of lung cancer cell line A549 in vivo and in vivo, and explore its mechanism. Methods:The most effective one as a target sequence was chosen from four Bmi-1 siRNA sequences which were designed by our lab, and one random sequence was chosen as a negative control. In short, the chemically synthesized siRNA and control sequences were connected to a retrovirus expressing vector, pSUPERretro-Neo plasmid, and then transfected into A549 cells. The stably transfected cells were cultured and passed. The level of mRNA and protein of Bmi-1 in A549 cells were assessed by RT-PCR and Western blot respectively. The proliferations of A549 cells in vivo was analyzed with MTT, trypan blue exclusion and plate colony forming methods. Flow cytometry was used for cell cycle analysis. The potency of tumorigenesis was observed in nude mouse through hypodermic inoculation of A549 cells. The expressions of cyclin D, p21/27, p-AKT and PTEN were analyzed by Western blot. Results:Compared to A549-ctr and A549-wt cells, Bmi-1 mRNA and protein levels all signiifcantly reduced in A549-Bmi-1-siRNA cells. Bmi-1-siRNA inhibited the growth, colony formation in vitro and tumorigenesis in vitro of A549 cells, and the interference cells cell cycle arrested in G1 phase. In A549-Bmi-1-siRNA cells, p-AKT and cyclinD1 expression were down-regulated while p21/p27 and PTEN were up-regulated. Conclusion:Silencing Bmi-1 gene inhibits the proliferation of A549 cells through G1 phase arrest, which involves the downregulation of cyclin D/p-AKT and upregulation of p21/p27/PTEN.
9.The Feasibility Study on the Application of Global Medical Device Nomenclature(GMDN).
Wanjuan YANG ; Jian ZHENG ; Jun LI ; Ying HUANG ; Chunqing ZHANG ; Jingli LI
Chinese Journal of Medical Instrumentation 2015;39(5):349-352
The article has analyzed the policy co-ordination, Industry coverage and technical experience of the global medical device nomenclature (GMDN) to our country, argued the feasibility on the application of GMDN in our medical device administration system, provided some reference on building the naming system of medical device in our country.
Equipment and Supplies
;
Feasibility Studies
;
Industry
;
Terminology as Topic
10.Expression and role of bone morphogenetic protein receptor Ⅱ in focal cortical dysplasia
Wei GUO ; Chunqing ZHANG ; Sixun YU ; Haifeng SHU ; Shiyong LIU ; Ning AN ; Hui YANG
Chinese Journal of Trauma 2011;27(5):472-476
Objective To detect the expression of bone morphogenetic protein receptor Ⅱ ( BMPR Ⅱ ) in human focal cortical dysplasia ( FCD Ⅱ b). Methods Fourteen specimens of FCD Ⅱ b surgically removed and pathologically verified were collected from June 2008 to June 2010 and the expression of BMPR Ⅱ in the normal brain tissues and the pathological specimens was detected by means of immunohistochemistry and western blot. Results In the normal brain tissues, BMPR Ⅱ was widely expressed in the cortical neurons of the grey matter, with no positive immunostaining in the white matter. In the cortical lesion of FCD Ⅱ b, BMPR Ⅱ was strongly expressed in the misshapen cells including balloon cells (BCs) , dysmorphic neurons (DNs) and giant neurons (GNs). Positive BMPR Ⅱ expression was also observed in the reactive astroeytes and low level expression of BMPR Ⅱ was found in the normal-appearing (NA) neurons. Western-blot analysis showed that BMPR Ⅱ expression tended to be lowered in the FCD Ⅱ b specimens compared with the normal brain tissues ( P < 0. 05 ). Conclusion The expression of BMPR Ⅱ is altered and reduced in the FCD Ⅱ b, suggesting that BMP signal pathway may participate in the pathogenesis of FCD.