1.Establishment of real time PCR for detecting plasma cell free DNA of rats and its significance.
Zhi-cheng GUO ; Lang YIN ; Xiao-hui WANG
Chinese Journal of Applied Physiology 2015;31(2):186-190
OBJECTIVEOvertraining is a serious problem in sports, assessed by comprehensive multi-index evaluation, but so far there is still no sensitive, specific monitoring indicator or simple evaluation method to evaluate it. This research established a method for detecting plasma cell free DNA (cfDNA) of rats by real time PCR and discuss edits significance: a new molecular marker of overtraining?
METHODSTwelve male SD rats were randomly divided into control group and overtraining group. The overtraining group rats were undertaken overtraining on a motor-driven treadmill for 5 weeks, while the control group rats kept quiescent. All the rats were drawn blood at pre-and after-5 weeks to detect plasma levels of cfDNA, testosterone (T) and corticosterone (Cort) as well as peroxidation/antioxidation parameters (T-AOC, MDA, SOD, GSH-Px) and creatin kinase (CK).
RESULTS(1) Plasma cfDNA of rat was detected specifically by our real time PCR. (2) Compared with control group rats, the plasma cfDNA of overtraining rats increased obviously (about 5.43 fold). (3) Plasma cfDNA was related to plasma T, Cort, T/C ratio and MDA (correlation coefficent were -0.729, 0.854, -0.655 and 0.720, respectively) rather than plasma T-AOC, GSH-Px, SOD and CK.
CONCLUSION(1) A real time PCR method was established successfully to determine plasma cfDNA of rat. (2) A remarkable raises of plasma levels of cfDNA were found in overtraining rats which were associated with T, Cort and T/C, suggested that plasma cfDNA might be a new molecular marker of overtraining. (3) The increase of plasma cfDNA of overtraining rat might correlate with enhanced oxidative stress induced by overtraining instead of muscle damage.
Animals ; Biomarkers ; blood ; Corticosterone ; blood ; DNA ; isolation & purification ; Exercise Test ; Fatigue ; blood ; Male ; Physical Conditioning, Animal ; Plasma Cells ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Testosterone ; blood
2.Proteomic study on effect of tangcao pill on microsome CYP450.
Li-Jun ZHANG ; Xiao-Fang JIA ; Lin YIN ; Xiao-Qian LIU ; Yin-Zhong SHEN ; Hong-Zhou LU ; Neng-Neng CHENG
China Journal of Chinese Materia Medica 2014;39(11):2136-2139
Tangcao pill is commonly applied in adjuvant and even alternative therapy for patients with AIDS. However, the herb contains complex ingredients, but with unknown effect against anti-HIV drug and unknown function. Because CYP450 emzyme is the main metabolic enzymes of the drug, it is of important significance to study the regulation of CYP450 enzymes before and after the combined administration of Tangcao pill and EFV. Proteomics, due to its high throughout and high sensitivity, has been widely applied in CYP450 enzyme study. In this paper, liver microsomes were separated through differential centrifugation. Their proteins were separated through SDS-PAGE. The three protein bands that CYP450 enzymes were located were cut and identified by liquid chromatography tandem mass spectrometry. Totally 16 CYP450 isoenzymes were identified. Furthermore, in order to make a quantitative analysis on the effect of tang herb on CYP450 emzyme, the multiple reaction monitoring (MRM) technology based on MS was adopted. The CYP2C11 was selected based on the results of the mass spectrum identification of proteins. The characteristic polypeptides were obtained through searching Expasy blast database. The m/z of the fragment ions was less than 800. In the paper, the m/z of ion pairs of CYP2C11 were 711.5/232.1, 711.5/319.2, 711.5/466.2 and 711.5/595.3, and the m/z of ESAT-6 (internal standard, IS) were 735.5/215.3, 735.5/389.3, 735.5/460.3 and 735.5/524.3. The relative peak (analyte/IS) area was adopted for the relative quantitative analysis. Compared with the EFV single administration group, the EFV and Tangcao pill combined administration group showed a 1.6-fold increase in CYP2C11. The results of the paper indicated that Tangcao pill may affect drug metabolism by regulating metabolic enzymes such as CYP2C11, but the specific mechanism still unknown.
Animals
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Cytochrome P-450 Enzyme System
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chemistry
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genetics
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metabolism
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Drugs, Chinese Herbal
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administration & dosage
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Electrophoresis, Polyacrylamide Gel
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Male
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Microsomes, Liver
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chemistry
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drug effects
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enzymology
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Proteomics
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Rats
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Rats, Sprague-Dawley
3.Learning resource exploration of anesthesiology postgraduate
Yu-qing WU ; Cheng-hua ZHOU ; Yong-ping WU ; Xiao-xing YIN ; Li-cai NG ZHA ; Yin-ming ZENG
Chinese Journal of Medical Education Research 2011;10(11):1296-1298
Thorough excavation and artful utilization of various kinds concrete and invisible learning resources contribute to the cultivation of excellent postgraduates.In postgraduate education of anesthesiology Xuzhou Medical College utilizes time,network,technique platform,research outcome,self-potentiality and clinical patients resources,which produces an active effect and has important instructional significance.
4.Ethical Inspection about laboratory animals.
Nai-bin YANG ; Xiao-jun PAN ; Jing-jing CHENG ; Jia-qiang LIN ; Jia-yin ZHU
Chinese Journal of Applied Physiology 2015;31(6):504-507
Laboratory animals and animal experiments are foundations and important support conditions for life sciences, especially for medical research. The animal experiments have drawn extensive attention from the society because of the ethical issue. This paper takes Wenzhou Medical University as an example to give a brief introduction to the ethical review about laboratory animals in the university so as to further draw attention and concerns from the public about the ethical issue of laboratory animals. We successively introduce its scientific projects, nurturing environment and ethical review of laboratory animals.
Animal Experimentation
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ethics
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Animals
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Animals, Laboratory
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Universities
5.Leptin activates STAT3 and ERK1/2 pathways and induces endometrial cancer cell proliferation.
Yi, LIU ; Liqun, LV ; Wei, XIAO ; Cheng, GONG ; Jie, YIN ; Donghua, WANG ; Hui, SHENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(3):365-70
Obesity is an established risk factor for endometrial cancer. Leptin, a secreted protein of the ob gene by white adipose tissue, plays an important role in the regulation of food intake and energy consumption in the brain and acts as a potential growth stimulator in normal and neoplastic cancer cells. However, a direct role for leptin in endometrial cancer has not been demonstrated. In the present study, the effect of leptin on the proliferation of Ishikawa endometrial cancer cells was investigated as well as the possible mechanism(s) underlying this action in endometrial cancers which express both short and long isoforms of leptin receptors. The expression of leptin receptor (ObRb) in Ishikawa cells was detected by RT-PCR and Western blotting. The cells after serum starvation, were treated by leptin with various concentrations (0, 10, 50, 100, 150 ng/mL) for different durations (6, 12, 24 h). The effect of leptin treatment on cell proliferation was examined by MTT assay. Meanwhile, inhibitory effect of Janus tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) inhibitor AG490 or extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor PD98059 on the proliferation of Ishikawa cells induced by leptin was also studied. Ishikawa cells were treated with 100 ng/mL leptin for various periods (0, 20, 40, 60 min), and the levels of STAT3 phosphorylation and ERK1/2 phosphorylation were examined by Western blotting. The results showed that leptin induced the phosphorylation of STAT3 and the activation of ERK1/2 in a time- and dose-dependent manner in the Ishikawa endometrial cancer cells. Blocking STAT3 phosphorylation with the inhibitor AG490, or blocking ERK1/2 activation by the specific ERK1/2 kinase inhibitor, PD98059, abolished leptin-induced proliferation of Ishikawa cells. In addition, leptin was found to potently induce the invasion of endometrial cancer cells in a Matrigel invasion assay. Leptin-stimulated invasion was effectively blocked by pharmacological inhibitors of STAT3 (AG490) and ERK1/2 kinase (PD98059). These results suggested that leptin promotes endometrial cancer growth and invasiveness by activating STAT3 and ERK1/2 signaling pathways and therefore blocking its action at the receptor level can be a rational therapeutic strategy.
6.Application study of malignant pancreas tumor treatment with embedment of ~(125)I by CT guide
Xiao-Kun HU ; Cheng-Bin YIN ; Shao-Kui WANG ; Yong-Kui WANG ; Jin JIN ;
Cancer Research and Clinic 2006;0(10):-
Objective To discuss the method,safety and effect of malignant pancreas tumor treated by CT guided percutaneous embedding of~(125)I. Methods 32 cases of malignant pancreas tumor with CT scan and contrast enhancement were retrospectively analysed.All the cases had been confirmed pathologically be- fore CT guided therapy.The number of~(125)I particle was 12~46,The distance between particles was 0.~1.2 cm. The number of puncture point was 1~2.The number of puncture direction was 2-5 times.Results Regarding 32 cases for 1 month,the size of the tumor reduced in 11 cases,no change in 20 cases,and increased in 1 case.As for 31 cases for 2 months,the size reduced in 16 cases,no change in 13 cases,and enlarged in 3 cases.Regarding 30 cases for 3 months,the size reduced in 18 cases,no change in 11 cases,and increased in 3 cases. Regarding 28 cases for 6 months, he size reduced in 10 cases, no change in 5 cases, and in- creased in 13 cases.Regarding 22 cases for 1 year,12 cases had been done the second therapy,the size of the tumor reduced in 16 cases,no change in 3 cases,and increased in 3 cases.Conclusion The size of pancreas tumors were reduced obviously,the symptoms were relieved after the treatment.The method turned out to be safe and accurate.
7.Experimental Study of Anxiety Disorder on Event-Related Potentials P_(300)
da-yuan, NAN ; xing-shi, CHEN ; zu-cheng, WANG ; ze-ping, XIAO ; hai-yin, ZHANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To study the characteristics of event-related potentials P300 in patients with anxiety disorder(AD). Methods P300 tests were carried out in 30 patients with AD and 30 healthy adult controls. ResultsPatients with AD had significantly delayed P3 latency ([326?16] ms vs [339?19]ms, P
8.Effectiveness of health education on iodine deficiency disorders and drinking-water-borne endemic fluorosis in Xinzhou City Shanxi Province in 2011
Jin-li, CHENG ; Jun-lan, ZHAO ; Yin, ZHI ; Xiao-li, YANG
Chinese Journal of Endemiology 2013;32(5):580-583
Objective To find out whether people in Xinzhou Shanxi know the hazard of both iodine deficiency disorders and drinking-water-borne endemic tluorosis,in Xinzhou Shanxi and to promote people actively participate in prevention of the diseases and to achieve a sustainable development of health education in endemic diseases.Methods Ningwu,Baode,Kelan and Jingle Counties were selected to carry out the.health education program of iodine deficiency disorders and Wutai,Xinfu and Fanshi counties were selected to carry out the health education project of drinking-water-borne endemic fluorosis in Xinzhou City Shanxi Province in 2011.To carry out the base line questionnaire survey,in the beginning and at the end of the project,three townships from each county were chosen,and one primary school was selected from each chosen township,15 housewives were selected from each chosen township and 30 fifth grade students were selected from each primary school.Results ① The baseline survey:a total of 366 pupils of grade 5 and 183 housewives were investigated,and the awareness rate of iodine deficiency disorders was 77.41% (850/1098) and 80.33% (441/549),respectively; a total of 270 pupils of grade 5 and 138 housewives were investigated,and the awareness rate of drinking-water-borne endemic fluorosis was 80.74% (654/810) and 81.40% (337/414),respectively; ② The effect evaluation:a total of 366 pupils of grade 5 and 181 housewives were investigated,and the awareness rates of iodine deficiency disorders were 91.62% (1006/1098) and 92.45% (502/543),which were compared with that of baseline investigation,and the awareness rates were improved significantly (x2 =84.69,34.04,all P < 0.01); a total of 270 pupils of grade 5 and 138 housewives were investigated,awareness rates of drinking-water-borne endemic fluorosis were 93.95% (761/810) and 93.48%(387/414),which were compared with that of baseline survey,and the awareness rates were improved significantly(x2 =63.94,27.47,all P < 0.01).Conclusions After implementation of health education project on endemic diseases,the self-protection awareness of the fifth grade students and housewives is promoted effectively,awareness of prevention knowledge on endemic diseases is raised,which lays a foundation for controlling and eliminating the endemic diseases.
9.Stimulation of bone marrow fibroblst conditioned medium on megakaryocyte in vitro and platelet in vivo.
Yan-hong HUANG ; Xiao-yin ZHOU ; Men-qun TAN ; Lamei CHENG ; Guangxiu LU ; Qiru WANG
Journal of Central South University(Medical Sciences) 2005;30(6):726-728
Animals
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Blood Platelets
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cytology
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Bone Marrow Cells
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cytology
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Cell Differentiation
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Cells, Cultured
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Culture Media, Conditioned
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Cyclophosphamide
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Female
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Fibroblasts
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cytology
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Male
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Megakaryocytes
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cytology
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Mice
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Stem Cells
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cytology
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Thrombocytopenia
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chemically induced
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therapy
10.Clinical Analysis of 317 Children with Fever of Unknown Origin
Li-Zhi CAO ; Ying-Chao WANG ; Xiao-Cheng YIN
Chinese Journal of Contemporary Pediatrics 2001;3(2):154-157
Objective To review the etiology of fever of unknown origin (FUO) and the methods used to establish a diagnosis. Methods A retrospective review of the medical records of 317 patients with FUO admitted between January, 1996 and December, 2000 was performed. Results Of the 317 children, 298 cases (94.0%) had a definitive etiology established. Of the 298 cases, 160 (53.7%) had infectious diseases and non-infectious diseases, such as collagen vascular disease, and neoplasm which accounted for a large fraction of the remainder. A diagnosis was established in 140 (47.0%) by comprehensive clinical analysis alone. Culture of bacteria in serum and biopsy specimens established a diagnosis in 64 cases (21.5%) and 37 cases (12.4%), respectively. Non-invasive imaging techniques (35 cases; 11.7%), autopsy (11 cases; 3.7%), bone marrow examination (6 cases; 1.9%) and retrospective diagnosis (5 cases; 1.7%) accounted for the remainder of the diagnosis. Conclusions Most cases of FUO can be diagnosed by clinical characteristics of the patients and by essential laboratory studies. Pathological examination is very important in diagnosing the etiology of FUO. Infectious diseases, collagen vascular disease and neoplasm are the major causes of FUO in children.