2.Inhibitory effects of PTEN gene transfection combined with L-OHP on proliferation of human cholangiocarcinoma cells
Ping CUI ; Tide DUAN ; Jian DONG ; Wei JIA ; Shupeng DAI
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To investigate the inhibitory effects of PTEN gene transfection combined with L-OHP on human cholangiocarcinoma cell line, QBC939, providing a new method for gene therapy of human biliary duct carcinoma. Methods: A eukaryotic expression vector containing PTEN gene was transfected into human QBC939 cells under mediation of lipofectamine and positive cell clones were selected and amplified. Expression of PTEN gene was detected by immunohistochemistry. MTT test was used to determine the in vitro activity of cells, electron microscope was applied to observe cell ultrastructure, and flow cytometry was used for determining the cell cycle and apoptosis. In vitro test was used to study the invasive ability of cells before and after treatment. Results: After transfected with PTEN gene, QBC939 cells had a higher expression of PTEN gene (P
3.The protective effect and its underlying mechanism of repetitive transcraniai magnetic stimulation on dopaminergic neurons in the substantia nigra of mice with Parkinson's disease
Qiaoyun DONG ; Quandong WANG ; Ping GU ; Li LIU ; Yanyong WANG ; Xiuqiao SUN ; Dongsheng CUI ; Mingwei WANG
Chinese Journal of Physical Medicine and Rehabilitation 2008;30(4):227-230
Objective To investigate the effect and its underlying mechanism of repetitive transcranial magnetic stimulation(rTMS)on dopaminergic neurons and glial cell line-derived neurotrophic factor(GDNF)in the substantia nigra(SN)in mice with Parkinson's disease(PD). Methods Thirty-two male C57 BL/6J mice were randomly divided into a normal saline group,a sham-rTMS group,a PD model group and an rTMS group,with 8 mice in each group. All the mice except those in the normal saline group were administered with 4 times of subcutaneous in jection of 1-methyl,4-phenyl-1,2,3,6 tetrahydropyridine(MPTP)15 mg/kg at 2-hour intervals in 1 day to induce neuronal injury in the SN and to establish acute mice PD model.The mice in the rTMS group received 5 trains of 1 Hz rTMS for 25 s,at the intensity of 1 Tesla(T)daily for 2 weeks.After rTMS,the effect of rTMS on PD mice was observed by immunohistoehemieal technique with regard to the expression of tyrosine hydroxylase(TH)and GDNF in the SN,and the quantitative analysis was performed by an advanced image-analysis system. Results Compared with normal saline group,the number of TH and GDNF immunoreaetive(TH-ir and GDNF-ir)cells and the corrected optical density(COD)values of PD model group and sham-rTMS group were significantly lower(P<0.01);Com pared with PD model and sham-rTMS groups,the numbers of TH and GDNF positive cells and COD values in rTMS group were significantly higher(P<0.05).There was a significant positive correlation between the count of TH-ir and that of GDNF-ir cells(r=0.836,P<0.01).The correlation between the COD values of TH-ir and that of GD-NF-ir cells was also significant(r=0.921,P<0.01).Conclusion rTMS markedly increased the number and the COD values of TH-positive dopaminergic neurons and simultaneously increased the number and the COD value of GD NF-ir cells in the SN of PD mice.These findings suggest that rTMS has neuroprotective effects on dopaminergic neurons in the MPTP-induced PD mice,which might be mediated by up-regulation of the expression of GDNF protein in the SN.
4.The protective effect of repetitive transcranial magnetic stimulation on the nigrostriatal system in Parkinson's disease
Quandong WANG ; Ping GU ; Qiaoyun DONG ; Yanyong WANG ; Dongsheng CUI ; Mingwei WANG
Chinese Journal of Physical Medicine and Rehabilitation 2012;(12):889-893
Objective To investigate the effect of repetitive transcranial magnetic stimulation (rTMS) on dopaminergic neurons in substantia nigra and nigrostriate fibers in the striatum in cases of Parkinson's disease (PD).Methods Forty male C57BL/6J mice were randomly divided into a control group,a PD model group,a sham-rTMS (srTMS) group and an rTMS group with 10 mice in each group.A PD model was established by subcutaneous injection of a solution of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP),and the mice were treated with rTMS or sham rTMS for 14 days.Tyrosine hydroxylase (TH) expression in the substantia and striatum were detected using an immunohistochemical technique,and the corrected optical densities (CODs) of TH in the striatum were analysed using an image analysis system.The Nissl bodies were detected by Nissl staining.The morphological disposition of nerve fibers in the striatum was detected using Warthin-Starry staining.Results The fraction of neurons expressing TH decreased significantly more in the substantia nigras of mice in the rTMS group than in the control group,the PD group and the srTMS group.In the control group the neural plasm of dopaminergic neurons was full of dark blue and granular Nissl bodies.Many Nissl bodies were lost in the PD and srTMS groups,and the remaining Nissl bodies were colored lightly.Few Nissl bodies were lost in the rTMS group.Positive TH reactions in the striatum were significantly decreased in the rTMS group compared to the control group.The average COD was also significantly lower.But positive TH reactions in the striata of the rTMS group mice were significantly greater than in the PD and srTMS groups,and the average COD was significantly higher.In the control group,the disposition of nerve fibers in the striatum was typically fasciculated,concentrated and ordered; in PD and srTMS groups,many fibers were lost,and the remaining nerve fibers were rare,ruptured and scattered ; in the rTMS group fewer nerve fibers were lost and the disposition of the remaining fibers was more fasciculated,concentrated and ordered than in the PD and srTMS groups.Conclusion rTMS may play a role in treating Parkinson's disease by protecting dopaminergic neurons as well as nigrostriate fibers and by improving the synthesis and transport of dopamine.
5.Expression of a begomoviral DNAbeta gene in transgenic Nicotiana plants induced abnormal cell division.
Xiao-feng CUI ; Yun-qin LI ; Dong-wei HU ; Xue-ping ZHOU
Journal of Zhejiang University. Science. B 2005;6(2):83-86
An increasing number of monopartite begomoviruses are being identified that a satellite molecule (DNAbeta) is required to induce typical symptoms in host plants. DNAbeta encodes a single gene (termed betaC1) encoded in the complementary-sense. We have produced transgenic Nicotiana benthamiana and N. tabacum plants expressing the betaC1 gene of a DNAbeta associated with Tomato yellow leaf curl China virus (TYLCCNV), under the control of the Cauliflower mosaic virus 35S promoter. Transgenic plants expressing betaC1 showed severe developmental abnormalities in both species. Microscopic analysis of sections of both transgenic and non-transgenic N. tabacum leaves showed abnormal outgrowths of transgenic N. tabacum to be due to disorganized cell division (hyperplasia) of spongy and palisade parenchyma. Immuno-gold labeling of sections with a polyclonal antibody against the betaC1 protein showed that the betaC1 protein accumulated in the nuclei of cells. The possible biological function of the betaC1 protein was discussed.
Cell Division
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physiology
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Cell Nucleus
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genetics
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metabolism
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ultrastructure
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Cells, Cultured
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DNA, Viral
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genetics
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Geminiviridae
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genetics
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Plant Diseases
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genetics
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virology
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Plant Leaves
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cytology
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genetics
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growth & development
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metabolism
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Plants, Genetically Modified
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growth & development
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metabolism
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Recombinant Proteins
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metabolism
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Tobacco
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cytology
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growth & development
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metabolism
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ultrastructure
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Viral Proteins
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genetics
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metabolism
6.Comparative study between the features of FFA and OCT in diabetic macular edema
Qin-Hui, WANG ; Jiu-Ping, LIU ; Dong-Mei, CUI ; Yan, HE ; Xin, TANG
International Eye Science 2014;(12):2210-2213
AlM: To assess the correlation between the features of optical coherencetomography ( OCT ) and fundus fluorescein angiography ( FFA) in diabetic macular edema ( DEM) .
METHODS: Totally 70 patients (135 eyes) with diabetic retinopathy ( DR) were evaluated by central vision, best corrected visual acuity ( BCVA ) , intraocular pressure, indirect ophthalmoscopy, slit lamp microscope combined+ 90D front mirror mydriatic fundus examination, mydriatic fundus color photography, OCT, FFA, the correlation between FFA and OCT were analyzed.
RESULTS: ln mild macular oedema cases, abnormalities in FFA was 56 eyes, abnormalities in OCT was 68 eyes (P=0. 0009);FFA showed 12 normal eyes, 10 eyes in OCT were characterized by diffused macular oedema; FFA was performed with cystoid macular oedema, OCT was 46. 7% with cystoid type .
CONCLUSlON: DME is diagnosed by Combination FFA with OCT, OCT is an indispensable tool when following up DME, and it has advantage in early application.
7.Applied study on optimizing therapeutic regimen of gastric cancer with the chemosensitivity test in vitro by MTT assay
Cui-Qing ZHANG ; Dong-Ping TANG ; Chuan-Min ZHANG ; Kai TANG ; Nan-Wu YANG ; Hai-Ping HE ;
Cancer Research and Clinic 2006;0(10):-
0.05. Conclusion The tumour chemosensitivity test in vitro gave some prediction and guidances for the clinic chemotherapy,and it could discover the drug resisting cases.The combined chemotherapy should be selected for gastric carcino- ma patients.
8.Investigation of transmission and protection in family of pulmonary MDR-TB
Yu-Sen CUI ; Ping WU ; Qi-Ping GE ; Ya-Dong DU ; Wei-Wei GAO
Chinese Journal of Modern Nursing 2010;16(12):1402-1404
Objective To investigate the situations of close contacting with MDR-TB in family and analysis the necessity and urgency of family protection.Methods Nine typical primary MDR-TB cases in our hospital that close contacted with family pulmonary TB patients.their relatives contacted with them and their treatment prognosis were retrospectively analyzed.Results All of the 9 cases were transmitted by lineal MDR-TB relatives.In one case,3 generations were pumonary TB patients.All of 4 family members were pulmonary TBpatients in another case.3 cases were treated with operations:2 cases with pulmonary lobectomy and 1 case with intestinal resection.Treatment prognosis:3 cases cured,2 cases not cured,4 cases dead.Conclusions Family close contacting with MDR-TB patients should be intervened earlier:including health education,effective protection and health examination regularly et al.
9.Study on identification of "Digeda" raw materials in Mongolian patent medicine by PCR amplification of specific alleles.
Zhan-hu CUI ; Xian-zhang HUANG ; Ping LONG ; Le ZHANG ; Dong-dong ZHAO ; Ying-li WANG ; Min-hui LI
China Journal of Chinese Materia Medica 2015;40(5):793-798
To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.
Alleles
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DNA Primers
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genetics
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DNA, Plant
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genetics
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Medicine, Mongolian Traditional
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Molecular Sequence Data
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Plants, Medicinal
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classification
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genetics
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Polymerase Chain Reaction
;
methods
10.Effects of inward rectifier potassium channel blockers on EPCs function.
Wen-ping LI ; Xiao-dong CUI ; Ning-ning HOU ; Xiao-yun ZHANG ; Jian-hua LIU ; Jing ZHANG ; Min CHENG
Chinese Journal of Applied Physiology 2015;31(5):448-451
OBJECTIVETo investigate the effects of inward rectifier potassium channel blockers (BaCl2, CsCl) on the functions of endothelial progenitor cells (EPCs).
METHODSDensity gradient centrifugation-isolated rat hone marrow mononuclear cells were cultured in vitro. EPCs were harvested and seeded on six culture dish when cells grew to 3-5 passages. Before testing the EPCs were synchronized with M199, which contain 2% fetal calf serum. In the end, EPCs were treated with different intervention. The experiment mainly included two parts: (1) BaCl2 (100 micromol/L) and free BaC2 of Tyrodes solution; (2) CsCl (1 mmol/L) and control. Cell pretreated with blockers above mentioned for 12 h, then the gene expression of stromal cell-derived factor-1 (SDF-1), epoprotenol (PGI2) were assessed, beyond that the ability of adhesion, migration were assayed with different tests. In addition, the medium was collected when EPCs were treated for 3 days. The levels of SDF-1 were measured by sandwich enzyme-linked immunosorbent assay (ELISA). Going even further, EPCs were treated with the signal pathway blockers in advance, after repeat the above steps, in order to analyze the change of SDF-1 and then discuss its mechanism.
RESULTSCompared with control group, BaCl2, CsCl could increase EPC adhesion and migration to same extent. Moreover, the gene expression of SDF-1, PGI2 was significantly up-regulated and the production of SDF-1 increased evidently. Furthermore, the mechanism of SDF-1 secretion increasing mainly was associated with eNOS signaling pathways.
CONCLUSIONBa2+ and Cs+ play important roles in increasing EPCs functions, such as adhesion, migration and secretion.
Animals ; Barium Compounds ; pharmacology ; Cells, Cultured ; Cesium ; pharmacology ; Chemokine CXCL12 ; metabolism ; Chlorides ; pharmacology ; Endothelial Cells ; cytology ; Enzyme-Linked Immunosorbent Assay ; Potassium Channels, Inwardly Rectifying ; antagonists & inhibitors ; physiology ; Rats ; Stem Cells ; cytology