OBJECTIVESTo explore intervention with electromagnetic noise for co-suppression effect on gap-junctional intercellular communication (GJIC) induced or strengthened by low intensity magnetic field with carcinogen 12-O-tetradecanoylphorbol-13-acetate (TPA).

METHODSFibroblast cells from NIH 3T3 mice were exposed to extremely low intensity magnetic field (MF) 0.2 mT, 0.2 mT + TPA or/and electromagnetic noise with the same intensity of MF for 24 h, and GJIC was determined using fluorescence recovery analysis after photobleaching (FRAP) with a laser-scanning confocal microscope (Leica, Germany).

RESULTSGJIC function could be co-suppressed by MF of 0.2 mT with TPA, with fluorescence recovery of (23 +/- 11)%, lower than that in the control group [(46 +/- 19)%] and in the group with TPA only [(34 +/- 17) %] (P < 0.01), indicating 0.2 mT MF plus TPA could co-inhibit GJIC (P < 0.01). Superposition of 0.2 mT noise MF could get a fluorescence recovery of (35 +/- 19)% and significantly antagonize its co-suppression by TPA.

CONCLUSIONElectromagnetic noise of 0.2 mT could block the intensifying effect of power frequency magnetic field on TPA-induced GJIC inhibition.

Animals ; Cell Communication ; drug effects ; physiology ; radiation effects ; Cell Line ; Electromagnetic Fields ; adverse effects ; Fluorescence Recovery After Photobleaching ; Gap Junctions ; drug effects ; physiology ; radiation effects ; Mice ; NIH 3T3 Cells ; Noise ; adverse effects ; Tetradecanoylphorbol Acetate ; pharmacology

In order to study the important factors involved in cationic liposome-mediated gene transfer, Lipofectamine 2000 or DOTAP was evaluated using three types of cells (Hep-2, MCF-7 and SW-480) in vitro transfection efficiencies. Different properties of the two reagents were analyzed and compared by DNA arrearage assay and MTT assay. Both Lipofectamine 2000 and DOTAP had strong capability to combine with DNA; Lipofectamine 2000 can get higher transfection efficiency of the three cells by using GFP as report gene, meanwhile, DOTAP can also get higher transfection efficiency against Hep-2 cell. However, DOTAP showed lower transfection efficiency against MCF-7 and SW-480 cell. On the other hand, the cytotoxicity assay showed that over 85% cell viability of MCF-7 cell could be achieved both by Lipofectamine 2000 and DOTAP under the optimal transfection condition. Relatively speaking, Lipofectamine 2000 has very high transfection efficiency in a broad range of cell lines, but because of the special selectivity of cell type on liposome, DOTAP also has a broad application prospect.
Cell Line, Tumor ; Cell Survival ; drug effects ; DNA ; genetics ; Fatty Acids, Monounsaturated ; chemistry ; toxicity ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Lipids ; chemistry ; toxicity ; Quaternary Ammonium Compounds ; chemistry ; toxicity ; Transfection

The cDNA of human FGF-21 was subcloned into the pSUMO expression vector and the fusion protein was induced to express in Escherichia coli Rosetta (DE3). The recombinant hFGF-21 was expressed in soluble form in the pSUMO expression system. The recombinant fusion protein was purified by Ni-NTA column. The purified recombinant protein was dialyzed against PBS for re-nature. To obtain pure and active recombinant protein, the fusion protein was subjected to cleavage with SUMO protease I. To examine glucose regulation activity of hFGF-21, 3T3-L1 pre-adipocytes were differentiated into adipocytes, glucose up-take activity of hFGF-21 was examined by glucose oxidase and peroxidase (GOD-POD) assay. Compared with no stimulation control, the recombinant hFGF-21 treatment led to a significant increase in glucose consumption of adipocytes and a significant decrease in concentration of glucose in the medium (P < 0.05, P < 0.001).
3T3-L1 Cells ; Adipocytes ; metabolism ; Animals ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Fibroblast Growth Factors ; genetics ; metabolism ; pharmacology ; Glucose ; metabolism ; Humans ; Mice ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Small Ubiquitin-Related Modifier Proteins ; genetics ; metabolism ; Solubility

Objective To explore the characteristics and components of recurrent papillary thyroid cancer in patients with metabolic syndrome .Methods We retrospectively reviewed the information of the patients with recurrent papillary thyroid cancer after initial surgery during January 1st ,2010 and December 31st ,2015 in the First Affiliated Hospital of Zhengzhou University .We compared tumor size ,lymph node metastasis ,recurrence time and postoperative invasion rate in metabolic syndrome and non-metabolic syndrome groups . Results Totally 82 patients were included with 34 men and 48 women .There was no significant difference between patients with and those without metabolic syndrome grouped by the classic diagnosis approach .However ,the lymph node metastasis grade of recurrent papillary thyroid cancer patients who also suffered from at least two metabolic disorders ,was lower ,especially in women (P=0 .002) .Moreover ,patients with metabolic disorders had shorter recurrence time (Pdiabetes=0 .034 , Pdyslipidemia =0 .037 , PBMI =0 .004 , PMetS2 =0 .036) .Conclusion Papillary thyroid cancer patients with metabolic disorder ,especially with two or more components of metabolic syndrome and overweight and/or obesity ,may have an increasing risk of recurrence .

AIM:To investigate the mechanism of angiotensinⅡ(AngⅡ)/angiotensinⅡ type 1 receptor (AT1R)pathway activating protein phosphatase 2A(PP2A)which leads to down-regulation endothelial nitric oxide syn-thase(eNOS)phosphorylation level in mesenteric arteries of rats.METHODS: METHODS: The mesenteric arteries of adult male SD rats(weighing 160~180 g;n=90)were isolated under aseptic conditions.Firstly,to determine the effect of angiotensinⅡdown-regulated eNOS(Ser1177)phosphorylation level,the mesenteric arteries were randomly divided into normal control(control)group and AngⅡgroup.The mesenteric arteries in AngⅡgroup were incubated with AngⅡat 1×10 -7mol/L,1×10 -6mol/L and 1×10 -5mol/L for 6 h,12 h and 24 h,respectively.Secondly,to investigate the mo-lecular mechanism by which angiotensinⅡ activated PP2A leading to down-regulation eNOS(Ser1177)phosphorylation level,the mesenteric arteries were randomly divided into control group, AngⅡ group and candesartan(CAN; a specific AT1R blocker)+AngⅡgroup.The mesenteric arteries were pretreated with 1×10 -5mol /L CAN for 1 h,then incubated with 1×10 -7mol/L AngⅡfor 12 h in CAN+AngⅡgroup.The protein levels of eNOS,p-eNOS(Ser1177),PP2Ac,p-PP2Ac(Tyr307)and protein phosphatase 2A inhibitor 2(IPP2A2 )in the arteries were determined by Western blot.The ac-tivity of PP2A in the arteries was detected by PP2A activity kit.RESULTS:Compared with the control group,the protein level of p-eNOS(Ser1177)in the mesenteric arteries was decreased after incubated with AngⅡfor 6 h,12 h and 24 h(P<0.05).The decreasing tendency of p-eNOS(Ser1177)showed concentration-dependently,especially in 12 h and 24 h groups.The expression of eNOS protein showed no significant difference in each group.Compared with the control group, the mesenteric arteries of the rats were incubated with AngⅡ at 1×10-7mol/L for 12 h in vitro, the protein levels of p-eNOS(Ser1177)were down-regulated(P<0.05); pretreatment with CAN significantly increased the protein level of p-eNOS(Ser1177)(P<0.05);the protein levels of eNOS showed no significant difference in each group.Compared with the control group,the protein levels of p-PP2Ac(Tyr307)and IPP2A2 were decreased after the mesenteric arteries were trea-ted with AngⅡat 1×10 -7mol/L for 12 h(P<0.05).Candesartan pretreatment restored the protein levels of p-PP2Ac (Tyr307)and IPP2A2 (P<0.05),however the expression of PP2Ac protein showed no significant difference in each group. Compared with the control group,the activity of PP2A was increased in the mesenteric arteries incubated with AngⅡat 1× 10-7mol/L for 12 h(P<0.05).Candesarten pretreatment inhibited the activity of PP 2A significantly(P<0.05).CON-CLUSION:AngⅡincreases PP2A activity via AT1R pathway,thus leading to down-regulation eNOS(Ser1177)phospho-rylation level in mesenteric arteries.The molecular mechanism of PP2A activation may be associated with decreasing the protein levels of p-PP2Ac(Tyr307)and IPP2A2.

OBJECTIVETo investigate if a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2 (MMP2) gene would inhibit the growth of hepatocellular carcinoma (HCC) in vivo.

METHODSUsing the recombinant adenoviral vector carrying antisense MMP2 gene (Ad-MMP2AS) which was constructed by us previously, to infect the human HCC cell line (Bel-7402). Then the invasiveness of the Bel-7402 cells was assayed in Matrigel, and the production of MMP2 in the Bel-7402 cells was detected with Western blotting analysis and Gelatin zymography. Then the Ad-MMP2AS-infected cells were subcutaneously inoculated in nude mice. After tumors developed, Ad-MMP2AS was injected intratumorally into pre-existing tumors. The tumors were removed, sectioned, and stained with H E.

RESULTSCompared with PBS or Ad-CMV-infected cells, the infected Bel-7402 cells with Ad-MMP2AS injections significantly reduced their MMP2 enzyme activity and invasiveness about 52.05% in Matrigel assays, and the tumor volumes in nude mice resulted in a 3.3-fold reduction. In addition, direct intratumoral injection of Ad-MMP2AS into pre-existing tumors significantly prevented further expansion of the tumor masses and resulted in a 63.06% reduction in tumor cell growth.

CONCLUSIONThe recombinant adenovirus with antisense MMP2 can effectively inhibit the invasiveness and growth of Bel-7402 cells in vitro and in vivo, and it has a therapeutic potential for HCC.

Adenoviridae ; genetics ; Animals ; Carcinoma, Hepatocellular ; pathology ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms, Experimental ; pathology ; Male ; Matrix Metalloproteinase 2 ; genetics ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Oligonucleotides, Antisense ; genetics ; Recombinant Proteins ; genetics ; Tumor Cells, Cultured

Objective To investigate the influence of combination of external prostaglandin and narrow band ultraviolet B (NB-UVB) on skin melanin and tyrosinase level in models with vitiligo.Methods Guinea pigs models with vitiligo were produced by 3％ hydroquinone cream.And then all the animal models were randomly divided into six groups according to the weight:normal group,model group,control group,medication alone group,light alone group and combination group.Normal group and model group were not treated.Control group was used the same amount of vaseline,2 times per day;the 0.25 mg · mL-1 dipropionazone tincture skin smear in medication alone group,2 times per day,stay on 2 months;the NB-UVB irradiation treatment instrument was exposed to 1 J · cm-1 at about 20 cm from the skin of the white spot,and the wavelength was 310 ～315 nm in light alone group,and once every other day,stay on 2 months;combination group,topical prostaglandina combination NB-UVB irradiation,frequency,method and time same to medication alone group and light alone group.After administration,the observation of vitiligo guinea pig skin tissue morphology and melanin distribution in the light microscope.Results The percentage of melanin hair follicles in model group,control group,medication alone group,light alone group and combination group were (37.50 ± 20.43) ％,(53.33 ± 14.38) ％,(55.00 ± 16.12) ％,(46.67 ± 19.66)％,(71.67 ± 7.53)％,respectively.The combined group is compared to the model group,the difference is significant in statistics (P ＜ 0.05).But comparison between control group,medication alone group,light alone group and model group,the difference were not significant in statistics (all P ＞ 0.05).Conclusion Combination of external prostaglandins and NB-UVB have some effect on vitiligo treatment.

BACKGROUNDCatheter ablation for atrial fibrillation (AF) has been demonstrated to be effective in a subsets of patients with AF. However, very few data are available in regard to patients with prior history of stroke undergoing catheter ablation. This study aimed to investigate the outcome of catheter ablation in AF patients with prior ischemic stroke.

METHODSBetween January 2008 and December 2011, of 1897 consecutive patients who presented at Beijing An Zhen Hospital for treatment of drug-refractory AF, 172 (9.1%) patients in the study population had a history of ischemic stroke. All patients underwent catheter ablation and were followed up to assess maintenance of sinus rhythm and recurrence of symptomatic stroke.

RESULTSAmong these 1897 patients, 1768 (93.2%) who had complete follow-up information for a minimum of six months were included in the final analysis. Patients in the stroke group (group I) and the no-stroke group (group II) were similar in regards to gender, body mass index (BMI), history of diabetes, type of AF, and left atrial size. The patients in group I were older than those in group II, and had a higher incidence of hypertension, chronic heart failure, lower left ventricular ejection fraction (LVEF), and higher CHADS2 scores. Six months after ablation, 107 (68.6%) patients in group I and 1403 (87.1%) in group II had discontinued warfarin treatment (P < 0.001). During a median follow-up of (633 ± 415) days, 65 patients in the group I and 638 in group II experienced AF recurrence, and five patients in group I and 28 in group II developed symptomatic stroke. The rates of AF recurrence and recurrent stroke were similar between group I and group II (41.7% vs. 39.6%, P = 0.611; 3.2% vs. 1.7%, P = 0.219; respectively).

CONCLUSIONCatheter ablation of AF in patients with prior stroke is feasible and efficient.

Aged ; Atrial Fibrillation ; surgery ; Catheter Ablation ; methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Stroke ; surgery

OBJECTIVETo investigate the relationship of sperm morphology with reproductive hormones in infertile men and the pathogenesis of teratozoospermia.

METHODSThis study included 90 infertile men aged 25 - 40 years. We measured their testis volumes using the Prader orchidometer, conducted routine semen analyses according to the WHO laboratory standard, and determined the concentrations of reproductive hormones and sex hormone-binding globulin (SHBG) by chemiluminescence and the levels of free testosterone (FT) and bioavailable testosterone (BioT).

RESULTSAll the subjects showed normal sperm concentration. Based on the results of semen morphology analysis, the 90 infertile men were equally divided into groups 1 (morphologically normal sperm <4%), 2 (morphologically normal sperm > or = 4% and <10%), and 3 (morphologically normal sperm > or = 10%), with no significant differences in age among the three groups (P>0.05). The volumes of the left testis were (14.27 +/- 3.65) ml, (16.90 +/- 3.57) ml and (14.57 +/- 3.57) ml, respectively (P = 0.006 group 1 vs group 2, P = 0.741 group 1 vs group 3, P = 0.014 group 2 vs group 3), and those of the right testis were (14.60 +/- 3.70) ml, (16.60 +/- 3.35) ml and (14.67 +/- 3.54) ml, respectively (P = 0.050). There were no significant differences among the three groups in prolactin, follicle-stimulating hormone, luteinising hormone, estradiol, total testosterone and SHBG, (P>0.05). The levels of serum FT were (0.25 +/- 0.07) nmol/L, (0.29 +/- 0.07) nmol/L and (0.31 +/- 0.13) nmol/L (P = 0.086 group 1 vs group 2, P= 0.010 group 1 vs group 3, P= 0.364 group 2 vs group 3), and those of BioT were (5.81 +/- 1.58) nmol/L, (6.78 +/- 1.55) nmol/L and (7.29 +/- 3.02) nmol/L, respectively (P = 0.086 group 1 vs group 2, P = 0.010 group 1 vs group 3, P = 0.364 group 2 vs group 3). The percentage of morphologically normal sperm was positively correlated with the levels of serum FT and BioT (P<0.05).

CONCLUSIONThe higher the levels of serum FT and BioT, the higher the percentage of morphologically normal sperm, which suggests that serum FT and BioT might be involved in the pathogenesis of teratozoospermia.

Adult ; Estradiol ; blood ; Follicle Stimulating Hormone ; blood ; Humans ; Infertility, Male ; blood ; physiopathology ; Luteinizing Hormone ; blood ; Male ; Prolactin ; blood ; Semen ; Semen Analysis ; Sex Hormone-Binding Globulin ; metabolism ; Sperm Count ; Spermatozoa ; abnormalities ; Testis ; Testosterone ; blood

OBJECTIVETo investigate the distribution of the single nucleotide polymorphisms (SNPs) in CAPN10 gene in Chinese population and their relation with type 2 diabetes mellitus in Han people of Northern China.

METHODSCAPN10 gene was sequenced to detect SNPs in 27 samples of different nationalities in China. 5 SNPs were genotyped with single-base extension (SBE) method to perform case-control study in 156 normal Han people of Northern China and 173 type 2 diabetes and the 3 positive loci reported in the article were performed haplotype analysis. One positive locus was also analyzed with transmission-disequilibrium test (TDT) and sib transmission-disequilibrium test (STDT) in 68 type 2 diabetes pedigrees (377 cases).

RESULTSA total of 40 SNPs were identified in length of 8,936 bp, with an average of 1 in every 223 bp; The SNPs in CAPN10 gene did not distribute evenly and the SNPs in Chinese was different from that reported in American Mexicans. There was no significantly statistical difference in the allele frequency of the 5 SNPs between case and control (P > 0.05), and the haplotype frequencies in the two groups were not much different (P > 0.05). There was no positive results in TDT and STDT analysis (P > 0.05).

CONCLUSIONSThe SNP distribution of CAPN10 gene varies with different nationalities. The studied SNPs in CAPN10 gene may not be the major susceptibility ones of type 2 diabetes mellitus in Han people of Northern China.

Alleles ; Asian Continental Ancestry Group ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; genetics ; Ethnic Groups ; Genetic Predisposition to Disease ; Genetic Testing ; Genotype ; Humans ; Polymorphism, Single Nucleotide