2.Bioinformatics analysis and construction of eukaryotic expression plasmid of Cx50 V64G mutation
Ping, LIU ; Ying, LIN ; Yue-Ying, YANG ; Jian-Qiu, ZHENG ; Ying, HOU ; Di, JIN ; Xiao-Bo, FU ; Hong-Mei, MA
International Eye Science 2007;7(5):1206-1208
AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.
3.Effects of iodine excess on TPO and NIS genes mRNA expression in rats
Jian-ying, XIA ; Jing-jing, SUN ; Xiu-mei, ZHANG ; Dan, YUE ; Cui-yao, WANG
Chinese Journal of Endemiology 2008;27(2):145-148
Objective To observe the effects of iodine excess on thyroid morphology,the expression of thyroid peroxidase and sodium iodide symporter mRNA and to explore their mechanisms.Methods One-month SD rats were divided into three groups:control iodine(CI),high iodine Ⅰ(HI Ⅰ)and high iodineⅡ(HI Ⅱ)and were fed with water containing iodine in different concentrations by adding K103(5,5000,10 000μg/L)respectively.Rats were sacrificed after being fed for six months.The morphology of thyroid was investigated under light microscopy and electron microscopy,the serum thyroid hormones and ratio of TPO/β-actin and NIS/β-actin were measured by radio-immunoassay and RT-PCR method.Results The major changes were increased follicles with colloid accumulation in HI groups.The levels of serum thyroid hormones TT3 and TT4 were decreased gradually from CI[(75.68±13.99,1.45±0.49)nmol/L]to HI Ⅰ[(73.82±16.48,1.34±0.31)nmol/L]and HIⅡ groups[(70.65±11.43,1.15±0.39)nmol/L],but there were no significant differences among three groups(F=O.371,l.163,P>0.05).The TPO and NIS mRNA expressions in HI Ⅰ(1.28±0.10,0.56±O.17)and HI Ⅱ(1.14±0.04,0.39±0.06)were significantly lower(F=30.863,62.62.675,P<O.05)than those of control group(1.39±0.08,0.71±0.13).Conclusions Chronic iodine excess leads to definite histological changes in rat thyroid,and inhibits the expressions of TPO and NIS mRNA as well as thyroid hormone synthesis,which in turn acts as a protective mechanism against iodine excess.
4.Effects of Pneumoperitoneum with Carbon Dioxide on Implantation and Growth of Tumor Cells
Guang-Yi WANG ; Xian-Ying MENG ; Jian-Hua GU ; Guo-Yue LV
Chinese Journal of Bases and Clinics in General Surgery 2003;0(02):-
Objective To study whether carbon dioxide used to establish pneumoperitoneum has an influence on port-site and intraperitoneal implantation and metastasis of tumor cells. Methods R 15 hepatic cancer cells were injected into 30 Wistar rats’ peritoneal cavities 1 hour before operation, then the 30 Wistar rats were randomly divided into 3 groups: gasless group, helium group and carbon dioxide group. The suspension was exposed to the gas environment for 2 hours, all animals were killed after 28 days and the port-site and intraperitoneal implantation and metastasis of tumor cells were examined.Results On port-site, intestinal serous coat, mesentery, greater omentum and diaphragm, the weights of tumor cells, in carbon dioxide group were (326.7?230.3) mg, (626.2?215.9) mg, (476.2?204.8) mg,(2 536.5?906.7) mg and (384.5?149.9) mg respectively; in helium group were (235.6?107.3) mg, (414.2?148.4) mg, (261.8?92.6) mg, (1 633.4?247.3) mg and(220.0?57.9) mg; in gasless group were (145.0?42.4) mg, (221.5?108.2) mg, (212.5?109.6) mg, (797.5?335.9) mg and 113.0 mg.The weights of carbon dioxide group showed a significant increase, compared with helium group and gasless group (P 0.05). Conclusion The insufflation of carbon dioxide promotes intraperitoneal tumor implantation and growth compared with helium and gaslessness in a rat model.
5.Relationship between Nitricoxide Synthase and Cell Apoptosis in Epilepsy
yue-ying, LIU ; xing-qiang, DENG ; zhuang-jian, XU ; rui, LI
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To explore the relationship of nitricoxide synthase(NOS)and cell apoptosis in epilepsy.Methods Pentylenetetrazol(PTZ)was used to build models of epilepsy.Cell apoptosis in each group were detected by flow cytometer,level of NOS was detected by colorimetric method.Results Levels of NOS increased obviously 1 hour after epilepsy and decreased;but at the 6th hour,NOS increased again and peaked at the 24th hour.Cell apotosis begin to increase at 6th hour and peaked at 24th hour after epilepsy,and decreased at 48th hour after epilepsy,there were significance differences compared with control group(Pa
6.The mechanism of androgen independent signaling pathway in castration-resistant prostate cancer and the research progress on related drugs
Meng-xia XUE ; Yue GU ; Jian-guo SUN ; Guang-ji WANG ; Ying PENG
Acta Pharmaceutica Sinica 2021;56(1):21-28
Prostate cancer is one of the common malignant tumors of male urogenital system, and the incidence of prostate cancer in China has increased significantly in the past decade. At present, endocrine therapy based on androgen blockade is the main method of clinical treatment except radical surgery and radiotherapy/chemotherapy for prostate cancer. However, the clinical benefit can only be obtained in the early stage of treatment, and nearly 90% of patients will develop to the castration resistance, and among them, nearly 90% of patients will have bone metastasis. The quality of life decreases sharply with the progression of disease for patients. In addition to the androgen signal pathway, studies have shown that many other oncogenic signal pathways have involved in the development of castration resistance, including classic cancer signaling pathways, immune and inflammatory signaling pathways, etc. Understanding the mechanism of androgen independent signal pathway in the formation of castration resistance will help to understand the off-target effect of androgen blocking therapy and introduce new treatment targets or strategies to get rid of the "no drug available" dilemma for clinical treatment of castration resistance.
7.Screen and Optimization of Cultured Medium Constituents for Biodegradation of ?-cypermethrin,a Synthetic Pyrethroid Insecticide,by Fusarium sp. Strain HG-P-01 via Central Composite Rotatable Design
Guo-Hua ZHONG ; Yue HE ; Jian-Jun LUO ; Shan GUAN ; Mei-Ying HU ;
Microbiology 1992;0(05):-
By means of comparing biomasses of biodegradation fungi,Fusarium sp.HG-P-01 for ?-cypermethrin,a synthetic pyrethroid insecticide used widely in China,in five different media,the Czapek-Dox medium was selected as the best medium for mycelia growth.Furthermore,an experiment of central composite rotatable design(CCRD) was used to optimize the content of nutrient components.The optimal composition of C,N and P in media for HG-P-01 were 20.94 g/L,1.82 g/L and 1.66 g/L,respec-tively,in which an expectant or real rate of ?-cypermethrin-degradation got to 96.34% or 93.78% by HPLC for a concentration of 50 mg/L after 24 h treatment.The predicted value in degradation rate model was con-sistent with that from HPLC method.
8.Effects of estradioi on radiation-induced apoptosis of bone marrow hematopoietic cells in mice
Bai-Long LI ; Jian-Ming CAI ; Jian-Guo CUI ; Ying-Song XIANG ; Fu GAO ; Ru-Jun YANG ; Yue-Cheng HUANG ;
Academic Journal of Second Military Medical University 1985;0(06):-
Objective:To investigate the effects of estradiol on ~(60)Co?-ray induced apoptosis of bone marrow hematopoietic cells of mice,and to discuss the related anti-irradiation mechanism.Methods:KM mice were randomly divided into 3 groups(15 mice/each group):control group(without radiation),pure radiation group and estradiol+radiation group(ER group).The pure radiation group was irradiated by 4.0 Gy?-ray at a dose rate of 1.15Gy/min;the ER group was administered with 0.1 mg estradiol(IM)at 10 days before 4.0 Gy?-ray radiation;and the control group received no special treatment.The apoptotic DNA segments of bone marrow hematopoietic cells were analyzed by DNA agarose gel electrophoresis;flow cytometry was used to examine the apoptosis rate of cells and expression of Fas and Bcl-2 at 4 h,8 h,and 12 h after irradiation.Results:Eight hours after radiation,the apoptotic DNA segments were obviously increased and apoptotic DNA ladder appeared,which was not seen in the other 2 groups.The apoptosis rate of bone marrow hematopoietic cells in ER group was significantly lower than that in the pure radiation group at 4,8,and 12 h after irradiation(P
9.Effect of formula of removing both phlegm and blood stasis on inflammatory reaction in Chinese mini-swine with coronary atherosclerosis.
Jian-Xun REN ; Lei LI ; Cheng-Ren LIN ; Jian-Hua FU ; Yue-Ying MA ; Jun-Mei LI ; Hong-Hai LI ; Min WANG ; Jian-Xun LIU
China Journal of Chinese Materia Medica 2014;39(2):285-290
OBJECTIVETo observe the effect of formula of removing both phlegm and blood stasis (TYTZ) in inhibiting the inflammatory reaction in Chinese mini-swine with coronary atherosclerosis.
METHODTotally 36 Chinese mini-swine were randomly divided to six groups: the normal control group, the model group, the Shujiangzhi group and TYTZ groups with does of 2.0, 1.0 and 0.5 g x kg(-1), and six each in every group. Except for the normal control group, all of other groups were fed with high-fat diet for 2 weeks. Interventional balloons are adopted to injure their left anterior descending artery endothelium. After the operation, they were fed with high-fat diet for 8 weeks to prepare the coronary atherosclerosis model. In the 8th week after the operation and administration, the intravascular ultrasound was adopted to observe the coronary artery plaque burden of each group and the pathological morphology of coronary artery. Such inflammatory factors as high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 were detected by ELISA. The expression of NF-kappaB p65 nuclear translocation was observed by the immunohistochemical method.
RESULTCompared with the normal control group, the model group showed significant increase in the coronary artery plaque burden at the end of the experiment (P < 0.01), notably abnormal structural changes in atherosclerotic vascular tissues, luminal stenosis, a large number of foam cells and inflammatory cell infiltration, remarkable growth of hs-CRP, TNF-alpha and IL-6 levels (P < 0.01). The immunohistochemical staining also showed the significant increase in the NF-kappaB p65 nuclear translocation of coronary artery of Chinese mini-swine in the model group. Compared with the model group, TYTZ could significantly attenuate atherosclerotic plaque burden (P < 0.01), inhibit the coronary luminal stenosis, reduce inflammatory cell infiltration, decrease such inflammatory cell factors as hs-CRP, TNF-alpha and IL-6 in serum, and inhibit the NF-kappaB p65 nuclear translocation of coronary artery (P < 0.05 or P < 0.01).
CONCLUSIONTYTZ can reduce the downstream inflammatory reaction by controlling NF-kappaB p65 nuclear translocation, so as to inhibit the occurrence and development of coronary atherosclerotic plaque in Chinese mini-swine.
Animals ; C-Reactive Protein ; metabolism ; Coronary Artery Disease ; blood ; complications ; drug therapy ; pathology ; Female ; Inflammation ; complications ; Interleukin-6 ; blood ; Male ; Medicine, Chinese Traditional ; methods ; Mucous Membrane ; drug effects ; secretion ; Swine ; Swine, Miniature ; Tumor Necrosis Factor-alpha ; blood
10.Effects of diazoxide on Fas/FasL protein expressions in rat myocardium suffered from long-term hypothermic preservation.
Ying FAN ; Ming-Zhi ZHENG ; Wei GUO ; Jian-Ping JIANG ; Li ZHU ; Yue-Liang SHEN ; Ying-Ying CHEN
Acta Physiologica Sinica 2008;60(1):11-16
The purpose of this study was to investigate the effect of a mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) opener, diazoxide (DE), on Fas/FasL protein expressions in rat heart suffered from long-term hypothermic preservation. The Langendorff isolated rat heart model was used. The hearts were stored in 4 °C Celsior solution with or without (control) DE for 8 h followed by 60 min of reperfusion. The recovery of rate-pressure product (RPP) was observed. Apoptotic cardiomyocytes were detected by TdT-mediated dUTP nick end labeling (TUNEL) technique. The expressions of Fas/FasL proteins were also analyzed by immunohistochemical method. The results showed that compared with the control group, DE (30 mmol/L) increased the recovery of RPP during reperfusion, reduced the percentage of apoptotic cells and the expressions of Fas and FasL proteins in rat hearts suffered from 8 h of hypothermic preservation. The above effects of DE were attenuated by a mitoK(ATP) channel inhibitor 5-hydroxydecanoate (5-HD). These results indicate that DE could alleviate rat myocardial injury induced by ischemia-reperfusion through reducing the expressions of Fas and FasL proteins via opening of mitoK(ATP)channel.
Animals
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Apoptosis
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Cryopreservation
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Decanoic Acids
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pharmacology
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Diazoxide
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pharmacology
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Fas Ligand Protein
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metabolism
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Heart
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drug effects
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Hydroxy Acids
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pharmacology
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Myocardium
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metabolism
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Myocytes, Cardiac
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cytology
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drug effects
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Potassium Channel Blockers
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pharmacology
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Potassium Channels
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Rats
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fas Receptor
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metabolism