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.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
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.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.
7.Changes of the cellular immune function of the elderly patients with nonsmall-cell lung cancer after chemotherapy
Gongping CHEN ; Ningfang LIAN ; Yongxu JIN ; Biying WANG ; Qichang LIN
Chinese Journal of Geriatrics 2011;30(11):941-943
ObjectiveTo investigate changes of the cellular immune function in the elderly patients with nonsmall-cell lung cancer (NSCLC) after chemotherapy.Methods T-lymphocyte subsets and natural killer cell were detected in 29 elderly patients with NSCLC,20 adults with NSCLC and 22 healthy elderly,and their levels were compared between pre-chemotherapy and at the end of 2 cycles of chemotherapy in the elderly patients with NSCLC.ResultsThe levels of CD3,CD4,CD8,CD4/CD8andNK cell were (58.9±15.8),(32.3±12.7),(22.0±9.8),(1.3±0.7),(21.6± 7.7),respectively in the elderly patients with NSCLC,(65.9 ± 7.2),(38.5 ± 7.6),(23.1 ± 9.2),(1.5±0.7),(16.8±6.2),respectively in adults with NSCLC and (67.3±9.0),(39.0±7.8),(23.9±9.3),(2.0±1.6),(22.5±5.8),respectively in healthy elderly.The levels of CD3 and CD4 were decreased (t=2.109,2.159,P<0.05) and NK cell was increased (t=2.273,P<0.05) while CD8 and CD4/CD8 had no difference(t = 0.406,0.736,P> 0.05 ) in the elderly patients with NSCLC as compared with adults with NSCLC.The levels of CD3,CD4,and CD4/CD8 were lower (t = 2.234,2.200,2.016,all P< 0.05) in elderly patients with NSCLC than in healthy elderly,with no significant change in the levels of CD8 and NK cell(t= 0.700,0.474,P>0.05) between the two groups.The levels of CD3 (51.6 ±10.3)was reduced(t=2.067,P<0.05) and CD4 (31.7 ± 11.7),CD8(21.6 ± 6.5),CD4/CD8 (1.3 ± 0.7),NK cell (26.0 ±12.7)had no remarkable difference (t =0.186,0.180,0.289,1.570,all P> 0.05)after chemotherapy in elderly patients with NSCLC.ConclusionsThe cellular immune function in the elderly patients with NSCLC is lower than in adults with NSCLC and healthy elderly,and further decreases after chemotherapy.
9.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.
10.Auditory tests in 14 patients(16 ears)with acoustic neuroma
Jin XU ; Chan LIU ; Xiuwu CHEN ; Nengjing LIAN ; Bo LIU
Chinese Archives of Otolaryngology-Head and Neck Surgery 2001;8(1):7-10
Objective:To investigate the relationship between hearing loss and acoustic neuroma(AN),the tests of pure tone audiometry,acoustic emissions impedance audiometry,audiometry brainstem response(ABR)and evoked otoacoustic emissions(EOAE)wee measured in 14 patients (16 ears)from March 1999 to December 2000.Methods:Fourteen patients (16 ears)with acoustic neuroma (8 males and 6 females,ranging in age from 21 to 72 years old)were diagnosed by CT or MRI scaning,and final confirmed by surgery and pathology.In the auditory tests ,efferent suppression test was curried out only in 4 ears with recordable emissions,promontory stimulation test (PST)was examined only in 5 ears with severe or profound deafness (hearing loss≥80dB SPL)who have no both measurable ABR and recordable EOAE.Results:It was found that 2 ears (12.5%,2/16)of the AN ears showed neural impairment,6 ears (37.5%,6/16)were cochlear impairment and 8 ears (50.0%,8/16)were cochlear-retrocochlear impairment.All of 4 tumors ears with EOAE emission have a disorders of efferent function.Conclusion:EOAE test had significant value for evaluation of the status of cochlear function (at the level of outer hair cells)in AN patients.The retrocochlear auditory nerve function of AN patients were evaluated by the tests of ABR combined PST which showed significant value.Results showed that the hearing impairment of AN have different levels of the peripheral auditory system according to auditory tests,including cochlear,eighth cranial nerve and efferent nerve level at the same or independently.