2.Cyr61 expression influences cancer cell proliferation and apoptosis via PI3K pathway in human ovarian carcinoma cells
Lan XIAO ; Chan HE ; Xin LI ; Ying CHEN ; Jiade ZHOU
Chinese Journal of Obstetrics and Gynecology 2012;47(8):616-620
Objective To investigate the relationship between cysteine-rich protein 61 ( Cyr61 ) and phosphatidylinositol 3-kinase ( PI3K ) signal pathway on cell proliferation and apoptotic in human ovarian carcinoma cells.Methods Recombinant human Cyr61 (rhCyr61) was pretreated with ovarian carcinoma cells.The expression of Cyr61 protein was detected by confocal spectral microscopy.Then treated the ovarian carcinoma cells with PI3K transduction inhibitors (LY294002) for 24 hours.Cell apoptosis was detected by flow cytometry (FCM).Cell viability was determined by methyl thiazolyl tetrazolium (MTT) method.The mRNA expressions of Cyr61,the protein levels of protein kinase B ( PKB),phospho-PKB and Cyr61 were assaved by real time-PCR and western blot analysis,respectively.Results The Cyr61 and phospho-PKB protein expression in two ovarian carcinoma cells (OV2008 and OVCAR-3 ) were increased in rhCyr61pretreated group.The decreasing of cell apoptosis [ ( 1.4 ±0.9)%,(2.1 ± 1.0)% ] and increasing of cell proliferation [ ( 124.0 ± 1.8)%,( 133.0 ±2.2)% ] was detected in the same time,compared with negative control group,there were significant difference ( P < 0.05 ).After exposed to LY294002 for 24 hours,the apoptosis rate of OV2008 and OVCAR-3 in pretreated with rhCyr61 group exposed to LY294002 were (21.1 ± 1.6)% and (26.4 ± 1.5 )%,respectively.Cells viability [ (59.0 ± 2.3 )%,(51.0 ± 2.0)% ]was also significantly decreased in OV2008 and OVCAR-3 pretreated with rhCyr61 cells.Meanwhile,the mRNA expressions of Cyr61 (3.2 ± 0.8,6.2 ± 1.1 ) and the protein levels of phospho-PKB and Cyr61 were greatly decreased.Compared with negative control group,there were significant difference in OV2008 and OVCAR-3 cells (all P < 0.0l ).Conclusions The activation of PI3K intracellular signaling pathways may lead to up-regulation of Cyr61 expression.Block PI3K signal pathway could significantly inhibit the expression of Cyr61,and may promote the apoptotic effects and inhibit the cell growth of ovarian carcinoma cells.
3.Progress in the clinical application of fiberoptic ductoscopy
Meng XIAO ; Fang XING ; Lan MU ; Xin WANG
Chinese Journal of Clinical Oncology 2014;(18):1199-1201
Conventional diagnostic methods used for pathologic nipple discharge (PND) include color ultrasound, mammary mo-lybdenum target X-ray radiography (mammography), nipple cytologic smears, and ductography. Diagnosis of PND through inspection yields indirect signs and has low positive rate. Fiberoptic ductoscopy (FDS) allows direct visualization of intra-ductal lesions, evaluates etiology of PND, and accurately locates intraductal lesions through wire marking. FDS is a valuable test for the early diagnosis of breast cancer and can help identify appropriate location for surgical excision. Interventional treatment for ductal ectasia and inflamma-tion is also efficient. Our study discusses FDS as a novel diagnosis and treatment method for PND patients.
4.Progress on the relationship between metformin and breast cancer
Lan MU ; Fang XING ; Meng XIAO ; Xin WANG
Chinese Journal of Clinical Oncology 2014;(21):1413-1415
Diabetes mellitus increases the risk of breast cancer and affects the prognosis of breast cancer patients. Metformin, an anti-diabetic drug, decreases blood glucose level and has a role in suppressing various cancers. In addition, metformin has a unique in-hibitory effect on breast cancer, i.e., it can inhibit breast cancer cells in vivo and in vitro. Moreover, metformin exerts its anti-tumor ef-fect on epidermal growth factor receptor 2 (HER-2) positive and monoclonal antibody trastuzumab-resistant breast cancer cell lines, breast cancer stem cells, and triple negative breast cancer cells. Metformin can reduce the risk of the breast cancer in patients with dia-betes, lower histologic grade, and increase expression of estrogen and progesterone receptors. Furthermore, metformin has certain effect on neo-adjuvant chemotherapy for breast cancer. This review aims to clarify the mechanism of metformin in restraining breast cancer based on basic and clinical study results.
5.The distribution of platelet glycoprotein I b? variable number tande repeat polymorphism and the relationship with cerebral infarction
Xiao-Min XIN ; Yu ZHAO ; Lan-Ying CUI ;
Chinese Journal of Laboratory Medicine 2001;0(02):-
Objective To study the distribution of variable number tande repeat(VNTR) polymorphisms of the platelet membrane glycoprotein Ⅰ b? in Han nationality at Harbin and the relationship between these polymorpbisms and cerebral infarction(CI).Methods The identification of alleles and genotypes of VNTR polymorphism of the glycoprotein Ⅰ b? gene was performed by polymerase chain reaction (PCR)in 200 healthy individuls and 200 CI patients(77 lacunar infarction patients and 123 atherosclerotic thrombotic infarction patients),to analyze The relationship between gene polymorphisms and cerebral infarction.Results(1)There were three types of alleles:B、C、D,and five types genotypes:BC,BD,CC, CD,DD in Harbin Han nationality.No person with A allele and BB genotype was found.(2)No statistically significant differences of GP Ⅰ b? gene VNTR polymorphism was found between CI patients or subtype CI patients and controls(P=0.412 and 0.572,respectively).Conclusions(1)This study indicates that the C and D alleles of VNTR polymorphisms of GP Ⅰ b? are the main alleles while the CC and CD genotypes are the main genotypes in Harbin Han people.(2)Our findings indicate that no association exists between the VNTR polymorphism of platelet GP Ⅰ b? gene and CI.
6.Impact of iodine deficiency on expression of thyroid stimulating hormone β splice variant in BALB/c mice
Xiao-hua, ZHUO ; Xin, LIU ; Zhi-mei, HU ; Xiao-yi, ZANG ; Yun, SUN ; Lan-ying, LI
Chinese Journal of Endemiology 2012;31(3):251-254
ObjectiveTo find out if the immune system derived thyroid stimulating hormone(TSH) β splice variant(TSHβ-Ⅴ) would be regulated by circulating thyroid hormone levels to get a further understanding of the function and mechanism of this TSHβ-Ⅴ in thyroid homeostasis.MethodsA total of 20 weaning Balb/c mice (half male and half female) were selected and randomly divided into two groups according to their body mass and gender(n =10).Mice of control group were fed with common diet and deionized water.Mice of the low-iodine(LI) group were fed with low-iodine diet(containing iodine 20 - 40 μg/kg,iodine-intake about 0.25 μg/d) and deionized water.The experimental period was 3 months.At the end of the experiment,mice were executed and the blood was collected to observe the levels of TSH and thyroid hormone by chemiluminescence immunoassay (CIA) ; bone marrow (BM),peripheral blood(PBL),thyroid gland and pituitary were collected to assay the TSHβ-Ⅴ mRNA expression by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR).ResultsThe serum free thyroxine(FT4) and total thyroxine(TT4) levels in LI group of mice[(0.47 ± 0.70)nmol/L,(2.41 ± 0.28)pmol/L] were significantly lower than that of the control group of mice [(55.2 ± 3.68) nmol/L, (32.72 ± 1.02) pmol/L,t =43.81,86.04 、all P < 0.01 ] and the serum total triiodothyronine(TT3) and free triiodothyronine(FT3) reduction in LI group of mice[ (0.76 ± 0.08)nmol/L,(4.01 ± 0.40)pmol/L] were significantly lower than that of the control group of mice [ (1.10 ± 0.06)nmol/L,(5.40 ± 0.38)pmol/L,t =9.81,7.5 1,P < 0.01 ].Iodine insufficiency strongly elevated the serum TSH in LI group of mice[ (35.67 ± 17.39)mU/L] than that in control group of mice[ (0.24 ± 0.10)mU/L,t =- 6.11,P < 0.01 ].The mRNA levels of TSH β-Ⅴ in BM (9.62 ± 0.60) and in PBL( 9.25 ± 0.83 ) of LI group of mice were lower than those in control group of mice (7.69 ± 0.36,7.11 ± 0.41,t =6.77,5.64,P < 0.01),while the mRNA level of TSH β-Ⅴ in pituitary of LI group of mice (1.99 ± 0.61) was increased compared with that in control group of mice (5.75 ± 0.98,t =- 8.02,P< 0.01).Compared with control group of mice(9.12 ± 0.62),the level of thyroid TSH β-Ⅴ mRNA in LI group of mice (9.32 ± 0.91 ) was not significantly changed (t =0.45,P > 0.05).There was no detectable native TSHβ in BM,PBL and thyroid.The mRNA level of native TSHβ in pituitary in LI group of mice( - 7.17 ± 1.78) was dramatically elevated compared to that in control group of mice( - 1.43 ± 0.51,t =- 7.60,P < 0.01 ).ConclusionsThe mRNA levels of TSHβ-Ⅴ are suppressed in BM and PBL in low iodinediet induced hypothyroidism mice,which suggest that immune system derived TSHβ-Ⅴ may be more important thannative TSHβ in immune-thyroid regulation.
7.The analysis of plasmid-mediated AmpC enzyme genotype and epidemiology of Escherichia coli and Klebsiella pneumoniae
Fu-Ying FENG ; Xiao-Peng LAN ; Xian-Yue YANG ; Ya-Bin ZHANG ; Xin-Lan HU ; Rong-Ying GUO ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To investigate the prevalence,genotype and epidemiology of plasmid- mediated AmpC enzyme of Escherichia coli and Klebsiella pneumoniae.Methods A total of 67 clinical isolates of nonrepetitive cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae collected by Fuzhou General Hospital and Fujian Provincial Hospital during a period of Sept.2004 to Mar.2005 were detected by three-dimensional extract test for AmpC enzyme,and PCR for AmpC enzyme and other ?-lactamase gene amplification and DNA sequencing were carried out for genotype of ?-lactamase.Plasmid transformation experiment was used to study the transfer of cefoxitin resistance.The homology of the isolates was determined by ERIC-PCR fingerprinting.Results At two hospitals in Fuzhou,the prevalence of plasmid-mediated AmpC enzyme among cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae were 16.7% and 10.5%, 8.0% and 0,respectively.Two isolates of Klebsiella pneumoniae produced DHA-1 plasmid-mediated AmpC enzyme,and 4 isolates of Escherichia cob and one strain of Escherichia coli produced CMY-2 and CMY-22 plasmid-mediated AmpC enzyme respectively.Furthermore,5 strains of Escherichia coli with CMY AmpC enzyme were also found simuhaneously to produce TEM-144,CTX-M-27,CTX-M-14 and TEM-1 ?-lactamase respectively.Three strains of Escherichia coli and one isolate of Klebsiella pneumoniae could transfer cefoxitin resistance to acceptant bacillus.ERIC-PCR fingerprinting reveals 2 strains of Klebsiella pneumoniae came from same clone,but 5 strains of Escherichia coli came from different clones.Conclusions The clinical isolates of Klebsiella pneumoniae producing DHA-1 plasmid-mediated AmpC enzyme and Escherichia coli producing CMY-2,CMY-22 plasmid-mediated AmpC enzyme are found in Fuzhou.CMY-22 AmpC enzyme and TEM-144 ?-lactamase are the first reported in the world,GenBank accession number: DO256079,DO256080
8.Screening analysis of 4 710 cases of inherited metabolic diseases in high - risk infants using urease pretreat-ment-gas chromatography-mass spectrometry
Hu HAO ; Sitao LI ; Xuejun OUYANG ; Suzhen FANG ; Hekui LAN ; Chunhua ZHANG ; Congcong SHI ; Xin XIAO
Chinese Journal of Applied Clinical Pediatrics 2015;(21):1660-1662
Objective To detect the incidence of inherited metabolic diseases(IMD)and disorders of metabo-lism in 4 710 high - risk infants,as well as providing basis of clinical diagnosis and treatment by using urease pretreat-ment - gas chromatography - mass spectrometry(UP - GC - MS). Methods Samples were collected from high - risk infants with IMD,after removing urea,putting in internal standard,removing protein,vacuum drying and bis (trimethyl - silyl)trifluoroacetamide / trimethyl - chlorosilane derivativing,UP - GC - MS was used to analyze compo-sitions such as organic acids,amino acids,carbohydrates,pyridoxines,purines and pyrimidines,then metabolic analysis was proceeded to refer to the normal detection value of the healthy children,finally a metabolic diagnosis was made ba-sing on the clinical data such as the high - risk clinical manifestations and general biochemical tests and other special examinations. Results In the 4 710 cases,there were 98 cases of IMD(2. 1% ),326 cases of suspected IMD(6. 9% ), 2 610 cases of metabolic disorders(55. 4% ). There were 98 cases of IMD,including 57 cases of methylmalonic aciduria,12 cases of propionic acidemia,7 cases of glutaric aciduria,5 cases of hyperphenylalaninemia,maple syrup u-rine disease and multiple carboxylase defects each,4 cases of isovaleric acidemia and 3 cases of 4 - hydroxy butyric acid urine disease. Conclusions UP - GC - MS is a effective way to diagnose IMD and metabolic disorders of infants. Common IMD in Guangdong Province include methylmalonic aciduria,propionic academia,glutaric aciduria,hyperphe-nylalaninemia,maple syrup urine disease and multiple carboxylase defects. The results of the tests can provide effective guidance for diagnosis and treatment of suspected infants.
9.Impact of setup errors on dose distribution of three dimensional conformal radiotherapy for patients with esophageal carcinoma
Chao GAO ; Lan WANG ; Zifeng CHI ; Chun HAN ; Jun WANG ; Xin ZHANG ; Guoxin MA ; Aiqin XIAO
Chinese Journal of Radiation Oncology 2009;18(4):270-273
Objective To measure the setup errors of patients with esophageal carcinoma during the treatment of three dimensional conformal radiotherapy (3DCRT), and to analyze the impact of setup errors on dose distribution of GTV,CTV and normal tissues around. Methods Forty-two patients with esophageal cancer treated by 3DCRT were included. The setup errors of each patient were measured once a week for 6 times by electronic portal imaging device (EPID). The setup errors were integrated into the treatment plan-ning system by moving the isocenter. Then the dose distribution of GTV, CTV and normal tissues were recal-culated. Results The systematic setup errors of the 42 patients were - 2.31 mm, - 0.55 mm and - 0.16 mm, and the random errors were 4.42 mm, 4.35 mm and 4.48 mm in the directions of lef-fight, anterior-posterior,and superior-inferior, respectively. The dose covered 95% GTV( D95 ) was reduced by 32 cGy and by 88 cGy for CTV D95. The lung V20 in the original plan and the integrated plan was 22.49% and 22.02%, respectively. The average dose of the heart in the two plans was 2077.62 cGy and 2036.23 cGy, respectively. In the original plan, no patient had maximum dose of spinal cord over 4500 cGy; While in the intergrated plan there were 18 patients had the spinal cord dose more than 4500 cGy, with a maximum dose of 5503.90 cGy. Conclusions The setup errors cause significant dose reduction of GTV and CTV, but not of the lung and heart . The maximum dose of the spinal cord may exceed 4500 cGy due to the setup errors.
10.Effect of travoprost on matrix metalloproteinase-2 expression in human ciliary muscle cells cultured in vitro
Yuqing LAN ; Jianhui XIAO ; Wei PENG ; Chi ZHANG ; Hui GUO ; Xin SONG
Chinese Journal of Tissue Engineering Research 2007;11(41):8414-8417
BACKGROUND:Travoprost can increase human ciliary muscle cell interspace, decrease uveoscleral outflow resistance and then decrease intraocular pressure. But whether this action pathway is conducted by enhancing the expression of matrix metalloproteinase (MMP) in the ciliary muscle cells remains unclear.OBJECTIVE:To observe the effect of travoprost on the expression of MMP-2 in the human ciliary muscle cells.DESIGN:Controlled observation analysis.SETTING:Zhongshan Ophthalmic Center,Sun Yat-sen University.MATERIALS:This study was Carried out in the Zhongshan Ophthalmic Center,Sun Yat-sen University between August 2005 and April 2006.Donor was from the unilateral eyeball of a youth patient,who was dead within one hour,had no any disease (informed consent was obtained from the relatives) in the Zhongshan Ophthalmic Center, Sun Yat-sen University.Rabbit anti.human MMP-2 polyclonal antibody (Boster Bioengineering Co.,Ltd,Wuhan),and travoprost (86610F,0.004% solution,ALCON company.USA) were used in this study.METHODS: Experimental intervention: 1μmol/L travoprost was added into bovine serum-free medium of human ciliary muscle cell, serving as experimental group,and meanwhile,the cells which were not interfered by drugs were taken as control group.In the experimental group,cells were harvested 6, 12,and 24 hours after travoprost being added.Experimental evaluation:MMP-2 gene and protein expressions in the human ciliary muscle cells in each group were detected by RT-PCR and ELISA methods.The activity of MMP-2 in each group was detected by Zymography technique.MAIN OUTCOME MEASURES:MMP-2 mRNA expression in the human ciliary muscle cell,MMP-2 protein expression and MMP-2 activity in the extracellular fluid.RESULTS:①In the experimental group, at 6,12 and 24 hours after travoprost being added,the relative expression of MMP-2 mRNA was gradually increased (F=236.959,P<0.01).②In the experimental group,at 6,12 and 24 hours after travoprost being added,MMP-2 protein was also gradually increased with time (F=38.110,P<0.01).③Zymography technique detection showed that in the experimental group,at 6,12 and 24 hours after travoprost adding,MMP-2 activity was gradually enhanced with time (F=74.348,P<0.01).CONCLUSION:After human ciliary muscle cell cultured in vitro being subjected to the intervention of travoprost.MMP-2 expression is gradually increased with action time of travoprost, and meanwhile MMP-2 activity is also gradually enhanced.