2.Study of EPO pretreatment's effect on expression of TNF-? gene in cultured cardiac myocytes with hypoxia/reoxygenation injury and the possible mechanism
Chuan QIN ; Yingbin XIAO ; Qianjin ZHONG
Chinese Journal of Thoracic and Cardiovascular Surgery 2003;0(03):-
Objective The present study is to investigate the effect of EPO pretreatment on TNF-? expression in cultured cardiac myocytes with H/R and to explore the possible NF-?B signal transduction mechanism. Methods The model of cultured cardiac myocytes with H/R was established and the cardiac myocytes were divided into 4 groups, including EPO group (treat with EPO 10?U/ml 24?h before H/R), EPO+PDTC group (treat with EPO 10?U/mL and PDTC 5??g/ml 24?h before H/R), PDTC group (treat with PDTC 5??g /ml 24h before H/R) and control group. Change of TNF-? gene expression before and after H/R in cardiac myocytes was detected with RT-PCR and western blot. Change of NF-?B activity before and after H/R in cardiac myocytes was assayed with EMSA. Results Before H/R, there was no significant difference in TNF-? mRNA and protein expression between the 4 groups and after H/R, TNF-? mRNA and protein expression increased significantly in the 4 groups compared to control group before H/R. After H/R, TNF-? mRNA and protein expression was lower in EPO group than in the other 3 groups. Before H/R, NF-?B activity was higher in EPO group than in the other groups. After H/R, NF-?B activity increased significantly in all the 4 groups compared to the control before H/R and NF-?B activity was lower in EPO group than in the other groups. Conclusion EPO pretreatment inhibited the upregulation of TNF-? gene expression after H/R in cardiac myocytes, which might be related to the inhibition of NF-?B activation; EPO pretreatment might inhibit the activation of NF-?B after H/R through the negative feed-back mechanism of NF-?B activation.
4.Treatment of primary liver cancer associated with active hepatitis.
Chao-hui ZUO ; Yong-zhong Ou YANG ; Sheng-chuan MO
Chinese Journal of Oncology 2006;28(4):319-320
Adult
;
Aged
;
Carcinoma, Hepatocellular
;
surgery
;
virology
;
Female
;
Follow-Up Studies
;
Glutathione
;
therapeutic use
;
Hepatectomy
;
methods
;
Hepatitis B
;
blood
;
drug therapy
;
Hepatitis B Surface Antigens
;
blood
;
Hepatitis C
;
drug therapy
;
Humans
;
Liver Neoplasms
;
surgery
;
virology
;
Male
;
Middle Aged
;
Prognosis
5.Evaluation of sentinel lymph node biopsy in detection of axillary lymph node metastasis in breast cancer.
Jian-Guo ZHANG ; Lei ZHONG ; Chuan-Le LI
Chinese Journal of Oncology 2007;29(3):235-236
Adult
;
Axilla
;
Breast Neoplasms
;
pathology
;
surgery
;
Carcinoma, Ductal, Breast
;
pathology
;
secondary
;
surgery
;
Carcinoma, Intraductal, Noninfiltrating
;
pathology
;
secondary
;
surgery
;
False Negative Reactions
;
Female
;
Humans
;
Lymph Node Excision
;
Lymph Nodes
;
pathology
;
surgery
;
Lymphatic Metastasis
;
Mastectomy
;
methods
;
Reproducibility of Results
;
Sentinel Lymph Node Biopsy
;
methods
;
Technetium Tc 99m Sulfur Colloid
6.cDNA Cloning of Maize Protein Kinase Gene ZmLRRPK1 and Its Expression Analysis
Ming-Di BIAN ; Chuan DENG ; Zhong-Yi BAI ;
China Biotechnology 2006;0(07):-
Abstract LRR-RLKs plays an important role in multiple signal transduction in plant.One full-length cDNA encoding a LRR-RLKs homologue was isolated from maize through in silico cloning and named as ZmLRRPK1(GenBank accession:EU873320).The predicted ZmLRRPK1 protein has 594 amino acids with an estimated molecular mass of 66kDa and an isoelectric point of 5.42.ZmLRRPK1 has the typical domain of LRR-RLKs.RT-PCR analysis indicated that ZmLRRPK1 expression was induced by ABA,mannitol and salt in coleoptiles of maize and kept high level during 24 hours.
7.Role of thrombus precursor protein in the monitoring of anticoagulation
Chuan QIN ; Yingbin XIAO ; Qianjin ZHONG ; Al ET ;
Chinese Journal of Thoracic and Cardiovascular Surgery 2003;0(04):-
Objective: To investigate the role of thrombus precursor protein (TPP) in the monitoring of anticoagulation after mechanical heart valve replacement. Methods: TPP and INR were compared between the coagulant group and control group. In the coagulant group, TPP and INR were also compared between the patients with atrial fibrillation and the patients without atrial fibrillation. The relationship between TPP levels and INR levels in 60 cases of anticoagulated patients was analyzed by linear regression. Results: It was found that the anticoagulant therapy could effectively decrease the level of TPP and increase the level of INR. In the anticoagulant group, the patients with atrial fibrillation had higher TPP level than other patients. No significant relationship was found between TPP levels and INR levels. TPP level in the patients with bleeding complications was far lower than 6 ?g/ml. Conclusion: TPP is a very valuable monitoring marker assisting INR. Patients with atrial fibrillation may require higher anticongulant intensity. INR and TPP should be tested at the same time in the patients receiving oral anticoagulation after mechanical heart valve replacement.
8.Expression levels of brain nicotinic acetylcholine receptor mRNA and protein in coal-burning type of fluorosis rats
Chuan-zhi, GUI ; Long-yan, RAN ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2011;30(3):239-242
Objective To observe the learning and memory changes in coal-burning type of fluorosis rats, detect the expressions of neuronal nicotinic acetylcholine receptor(nAChR) at mRNA and protein levels in rat brains and to reveal the mechanism of changed learning and memory ability. Methods Twenty-four healthy SD rats, weighting 100 - 120 g, were randomly divided into three groups(8 in each). Control group was fed with normal diet, and low- and high-dose fluoride groups were fed with corn polluted with high fluoride (fluoride were 11.30,104.20 mg/kg, respectively) during drying processes with local burning-coal from the areas of endemic fluorosis to established rat model of chronic fluorosis. After exposed to fluoride for 6 months, behavioral changes were measured by Morris water maze. Animals were sacrificed, the brain was taken, after homogenizing the fluoride content of brain tissue was determined by fluoride ion selective electrode. The α3, α4 and α7 nAChR subunits at mRNA and protein levels were analyzed by real-time PCR and Western blotting, respectively. Results For rats in low- and high-fluoride groups, the escape latency time[(12.42 ± 8.03),(17.48 ± 8.05)s] was significantly longer than that in the control[(7.04 ± 3.29)s, all P< 0.05]. For rats in high-fluoride group, the numbers of crossing the platforms (1.62 ± 0.87) and the time of staying at the platforms[(16.70 ± 5.02)s] were significantly decreased as compared to that of control[3.53 ± 1.67, (23.33 ± 5.35)s, all P < 0.05]. The fluoride content in rat brain tissue in low- or high-fluoride groups [(1.14 ± 0.04), (1.79 ± 0.04)mg/kg] was significantly higher than that of control [ (0.52 ± 0.05) mg/kg, all P < 0.05]; in addition, the amount of fluoride in brain tissue of high-fluoride group was significantly higher than that of low-fluoride group(P < 0.05). In high-fluoride group, the mRNA expressions of α3, α4 and α7 nAChR subunits in rat brains(1.51 ± 0.20,1.45 ± 0.06,1.63 ± 0.08) were significantly lower as compared to controls (1.79 ± 0.11,1.66 ± 0.14,1.83 ± 0.06, all P< 0.05); whereas there were no significant changes in mRNA levels of these receptor subunits of the rat brains between low-fluoride group(1.65 ± 0.17,1.59 ± 0.09,1.71 ± 0.03) and controls (all P > 0.05). Furthermore, the protein levels of α3, α4 and α7 nAChR subunits in rat brains of highfluoride group(0.58 ± 0.13,0.16 ± 0.03,1.41 ± 0.38) and low-fluoride group(0.56 ± 0.23,0.08 ± 0.02,0.51 ± 0.16) were significantly lower than those of controls( 1.48 ± 0.42,0.57 ± 0.21,2.56 ± 0.26, P<0.05 or < 0.01). Conclusions Decreased ability of learning and memory in coal-burning type of fluorosis rats may be associated with declined expressions of nAChR at proteins and mRNA levels, which might be the main mechanism of the behavior change.
9.Expression of Apoptosis Associated Proteins Survivin and Fas in Nephroblastoma and Their Clinical Significance
zhong, JIANG ; chuan-min, YANG ; bu-xian, JIANG
Journal of Applied Clinical Pediatrics 2004;0(11):-
Objective To explore the role of Survivin and Fas in the tumor cells′ invasion and metastasis and to evaluate whether they can be used as a reliable predictor for diagnosis and a new treatment pattern for therapy.Methods Thirty paraffin-embeded material of nephroblastoma were collected.The control group was composed of 15 specimens of normal kidney tissue acquired from paratumor region of the material.Immunohistochemistry methods was used to detect the stain of Survivin and Fas proteins in normal kidney and nephroblastoma specimens,compared them with clinical stage,pathology type and the outcome.Results Positive expression rate of Survivin in tumor specimens and significantly higher than that in control group(53.3% vs 0,P
10.Application of High Resolution Melting Method in the Detection of GLP1R Gene Polymorphism Site rs3765467
Yuan ZHANG ; Xia HE ; Lei ZHONG ; Junlan CHUAN ; Enwu LONG
China Pharmacy 2017;28(17):2305-2308
OBJECTIVE:To establish the method for the detection of the known glucagon-like peptide 1 receptor (GLP1R) gene mutation site rs3765467(NT_007592.16,position:39065819),and to evaluate its accuracy and practicability. METHODS:Peripheral venous blood samples of 72 healthy subjects were collected in medical examination center of our hospital during Oct. 2015-Feb. 2016. The whole blood DNA was extracted by column extraction method. After amplified by touch down PCR,high reso-lution melting(HRM)method was adopted to analyze amplified product. Sequencing verification by double stranded chain termina-tion method(Sanger sequencing method)was performed for 38 test samples. The results of 2 methods were compared. RESULTS:The results of mutation scanning showed that there were 39065817 and 39065819 polymorphism sites in amplified segments. Four types of mutations were detected by HRM method [GCG/GCG,GCA/GCG or ACG/GCG,GCA/GCA or ACG/ACG,A(G) CA(G)],but 6 types of mutations was detected by Sanger sequencing method [GCG/GCG,ACG/GCG,ACG/ACG,A(G)CA (G),GCA/GCG,GCA/GCA]. CONCLUSIONS:HRM method can identify GCG/GCG and A(G)CA(G)genotype,but can not identify GCA/GCG and ACG/GCG heterozygous mutation,GCA/GCA and ACG/ACG homozygous mutation. The method is not suitable for the detection of single nucleotide polymorphism for multiple neighboring sites. In the detection of single nucleotide mu-tation,economical and simple method should be selected after comprehensive analysis of sequence.