2.Effect of lipoxin A4 on the expression of aquqporin-1,3,5 in the typeⅡpenumonocyte of rat
Hongxia MEI ; Shengwei JIN ; Qiogquan LIAN
Chinese Journal of Emergency Medicine 2009;18(4):406-411
Objective To study the effects of Lipoxins A4(LXA4)on the expressions of aquaporin(AQP)1,3,5 in type Ⅱ pneumonocytes(ATⅡ)of rat treated with lipopolysaccharide(LPS).Method One pathogenfree male Spree Dawley(SD)rat every time.weighing 200~250 g,were used for the study.The typeⅡpenumonocytes of rats were isolated and purified,and the changes of cellular ultrastructure were observed by electron microscope in order to get the purity quotien>90%.The type Ⅱ pneumonocytes were divided randomly into five groups,namely,vebicukun group(alcohol 0.7μL/mL),control group,LXA4 group(1×10-7mol/mL),endotoxin group(LPS 1μg/mL)and LXA4+LPS group(LXA4 1×10-7mol/mL,LPS 1μg/mL).AQP-1,3,5 mRNA of in the typeⅡpenumonocytes were assayed by using reversal transcription poly chain reaction(RT-PCR),and the expressions of AQP-1,3,5 protein were detected by using.immunohistochemistry(IHC).One each specimen,these tests were repeated for six times.ANOVA was used for statistical analysis.Results RT-PCR and IHC showed that when AT Ⅱ treated with 1 μg/mL LPS for 4 hours,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were significantly decreased in LPS group compared with control group(P<0.01).However,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein after application of LXA4 significandy increased in LPS+LXA4 group in comparison with LPS group(LPS+LXA4,AQP1:0.647±0.132,AQP3:0.900±0.856,AQP5:0.879±0.058;LPS,AQP1:0.297±0.133,AQP3:0.512±0.113,AQP5:0.647±0.110;P<0.01).The AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were aignificandy increased in LXA4 group in comparison with control group(LXA4,AQP1:0.539±0.142,AQP3:0.818 4-0.176,AQP5:0.841±0.066;Blank Control,AQP1:0.518±0.139;AQP3:0.138±0.136,AQP5:0.766±0.066;P<0.01).Conclusions AQP-1,3,5 exist in typeⅡpenumonoeyte of rata,and the LXA4 can up-regulate the mRNA and protein expressions of AQP-1,3,5 in Type Ⅱ penumonocytes of rats treated with LPS.
3.Laparosocopically assisted treatment of lower digestive tract hemorrhage in children
Xingshuo JIN ; Hong LIU ; Shuhua LIAN
Chinese Journal of Minimally Invasive Surgery 2005;0(08):-
Objective To explore the value of laparoscopic diagnosis and treatment for Meckel’s diverticulum in children. Methods Laparoscopic exploration was performed in 11 children with suspected intestinal Meckel’s diverticulum. In other 2 children, the diverticulum was encountered during laparoscopic appendectomy, and then the umbilical incision was prolonged to take out the corresponding intestine for resection. Results No conversions to laparotomy were required in all the 13 children. The operation time lasted 30~90 min (mean, 75 min). The children were discharged at 5~6 postoperative days without complications. Conclusions Laparoscopic diagnosis and treatment for Meckel’s diverticulum in children is effective.
4.Amplification of Full-length mRNA 5′Ends Based on Template-switching Effects
China Biotechnology 2006;0(12):-
"CapFinder" technology,which can be used to clone the full length of 5′ UTR sequence of mRNA,was described.This technology used the terminal transferase activity of certain MMLV RT variants that added 3-5 residues(predominantly dC) to the 3′end of the first-strand cDNA exhibited when MMLV RT reached the 5′cap structure of mRNA.In the reverse reaction system containing GGG oligo,the terminal transferase activity was harnessed by the GGG oligo whose terminal stretch of dG residues can anneal to the dC-rich cDNA tail and serve as an extended template for RT.After RT switch templates from the mRNA template to the GGG oligo,a complete cDNA copy of the original RNA was synthesized with the additional GGG oligo sequences at the end.5′UTR of mRNA can be amplified with GGG oligo as forward primer and a gene-specific reverse primer.5′UTR of Bt toxin receptor E-Cadherin gene in midgut of cotton bollworm was cloned.
5.Effects of siRNA targeting PCNA gene on nasopharyngeal carcinoma CNE2 cell cycle
Bing LIAN ; Jiqun WANG ; Lin JIN
Chinese Journal of Pathophysiology 2000;0(08):-
AIM:To explore the effects of small interfering RNA(siRNA) targeting PCNA gene on nasopharyngeal carcinoma CNE2 cells growth and cycle.METHODS:Three synthesized siRNA targeting PCNA gene was transfected into CNE2 cells by using LipofectamineTM reagent.The PCNA mRNA and PCNA protein were detected by real-time polymerase chain reaction(RT-PCR) and immunohistochemical method.Inverted phase contrast microscope was used to determine the CEN2 cells growth before and after PCNA-siRNA transfected.Flow cytometry was used to observe the cell cycle.RESULTS:In CNE2 cells after PCNA-siRNA transfection,the expressions of PCNA mRNA and protein were down-regulated at different degree.Inhibition ratio of PCNA mRNA was 98.5%.Meanwhile,the cell cycle was suffocated at G0/G1 stage.CONCLUSION:The synthesized PCNA-siRNA effectively interferes nasopharyngeal carcinoma cells by down-regulating the expressions of the PCNA mRNA and its protein,therefore inhibits the growth of CNE2 cells.Future application of PCNA-siRNA in the gene therapy of nasopharyngeal carcinoma might be expected.
6.Effect of Copper and Cu/Zn Ratio in Blood on Blood-fat in Copper Deficiency Rats
Lian DUAN ; Zhen WU ; Yinlong JIN
Journal of Environment and Health 1992;0(05):-
Objective To study the relation between copper,copper-zinc ratio in blood and blood-fat only in the copper intake lower than the normal acceptable daily intake,and to provide a unified dosage evidence for copper multi-ways exposure.Methods Forty-two SD rats were divided into 7 groups randomly.The rats in one group among them as the normal control group were fed on the normal fodder and water,the other 6 groups were fed on special fodder without copper and deionized water and i.g.copper gluconate at doses of 0ADI(the acceptable daily intake of rat),1/625ADI(0.000 128 mg/ml),1/125ADI(0.000 64 mg/ml),1/25ADI(0.003 2 mg/ml),1/5ADI(0.016 mg/ml) and ADI(0.8 mg/ml),for 30 days.The activity of ceruloplasmin(CP),the content of copper,zinc in the blood and the level of total cholesterol(TC),triglyceride(TG),high-density lipoprotein(HDL) and low density-lipoprotein(LDL) were determined.Results In the case of copper intake lower than the normal acceptable daily intake,the activity of CP in the blood of all treated rats were lower than the normal level(P
8.The expression of α-enolase in hypertrophic cardiomyocytes induced by endothelin-1(ET-1)and regulation mechanism for the expression
Lian ZHU ; Ningyuan FANG ; Xian JIN ; Haiya WANG ; Pingjin GAO
Chinese Journal of Geriatrics 2008;27(11):851-855
ObjectiveTo investigate the protein expression of ERK1/2,p-ERK1/2,HIF-1α and α-enolase in hypertrophic cardiomyocytes induced by ET-1 and explore the regulation mechanism of overexpression of α-enolase in hypertrophic cardiomyocytes.MethodsET-1-induced abnormal cardiomyocytes were used as model of cardiac hypertrophy.Cellsurface area, [<'3>H]-leucine incorporation and the actin staining were measured to determine the extent of hypertrophy. Cultured cardiomyocytes were divided into 4 groups at random, control group, PD98059 treated group, ET-1 treated group and PD980594- ET-1 treated group. The protein expressions of ERK1/2, p-ERK1/2,HIF-1α and α-enolase were detected by immunoblotting analysis.ResultsCompared with the control group , cell surface area and [<'3>H] leucine incorporation were increased in ET-1 treated group ((1350.7±107.5)μtm<'2> vs. (896.1±70. 2)μtm<'2> , P<0.05; (1387.9±14.8) dpm vs. (787.7±10.2)dpm,P<0.013. Actin staining showed that ET-1-treated cardiomyoeytes had more intense actin staining and clear cross-striations than did control group, which suggested that myocardial cell hypertrophy could be induced by ET-1 in WKY neonatal cardiomyocytes. After MEK 1/2 inhibitor PD98059 was used, the cell surface area and [<'3>H] leucine incorporation were decreased in PD980594-ET-1 treated group compared with ET-1 treated group[(907.0±92.5)μm<'2> vs. (1350.7±107.5)μm<'2>;(841.5±10. 5)dpm vs. (1387.9±14.8)dpm, both P<0.05], which suggested that myocardial cell hypertrophy could be regulated by ERK1/2 signal pathway. Immunoblotting analysis showed that the protein expressions of p-ERK1/2, HIF-1α and α-enolase increased after ET-1 treatment,while PD98059 as an inhibitor of the upstream kinase of ERK1/2 was used, the protein expressions of HIF-1α and α-enolase were partially inhibited.ConclusionsET-1 induces hypertrophic cardiomyocytes through ERK1/2 phosphorylation in cultured neonatal rat cardiac myocytes.ERK1/2 and HIF-1α signal pathway may play an important role in the overexpression of α-enolase in the hypertrophic cardiomyocytes.
9.Expression of intercellular adhesion molecule-1 and lymphocyte function-associated antigen-1 in salivary glands of patients with Sj(?)gren's syndrome
Jin-Hua SHI ; Yun-Xia WU ; Lian-Ping YANG ;
Chinese Journal of Rheumatology 2001;0(05):-
Objective To explore the role of intercellular adhesion molecule-1(ICAM-1)and lym- phocyte function-associated antigen-I(LFA-1)in the pathogenesis of Sjgren's syndrome(SS)and provide a theoretical basis for clinical therapy.Methods Immunohistochemical method was used to detect these two cellular adhesion molecules in labial salivary glands of primary Sjgren's syndrome patients and 15 healthy controls.Semiquantitative analysis was performed by image analysis software.Results①In salivary gland samples,the expression of both ICAM-1 and LFA-1 was significantly higher compared to that of controls(P
10.Inhibitory effect of SIN-1 on ICAM-1 expression in rat kidney after ischemia/reperfusion injury
Hong LIU ; Xiongfei WU ; Xiyu JIN ; Lian LI ;
Journal of Third Military Medical University 2003;0(09):-
Objective To explore the therapeutic mechanisms of nitric oxide donor (SIN 1) in ischemic acute renal failure (IARF). Methods The effects of SIN 1 on intercellular adhesion molecule 1 (ICAM 1) expression and inflammatory cell infiltration in rat kidney after ischemia/reperfusion (I/R) injury were examined using ICAM 1, ?2 integrin polyclonal antibody, and immunohistochemistry. The renal functions were measured simultaneously. Results Progressive increased expression of ICAM 1 was found in kidneys, especially in vasa recta, after IR injury. ?2 integrin positive cells increased simultaneously in the outer medulla. The two indexes reached the peak values at 24 h after I/R injury. SIN 1 infusion at the beginning of reperfusion could remarkably inhibit the enhanced ICAM 1 expression, reduce the local infiltration of inflammatory cells, and ameliorate the renal functions. Conclusion NO donor can inhibit the renal expression of ICAM 1 and the infiltration of leukocytes after ischemia reperfusion injury, which may play an important role in the treatment of ischemic acute renal failure.