Objective: To investigate the detection ability of two reagents by automated coagulation analyzer ACL-TOP using immune turbidimetric method, and to determine the merits and demerits of them. Methods:Two reagents were compared in the study. We selected 532 cases of hospitalized patients in Anzhen Hospital, 388 cases of which were unstable angina, 60 cases were pulmonary thromboembolism, 70 cases were coronary atherosclerosis, 14 cases were abdominal aneurysm, and the rest were the patients with high blood pressure and coronary heart disease. The plasmas of all the patients were extracted, and D-dimer was detected on ACL-TOP by D-Dimer (referred to as reagent A) and D-DHS (referred to as reagent B). Results:The results of D-dimer increased in different degrees, and there are 88 cases of false positive cases. The case whose FDP and individual D-dimer of D-dimer simultaneous detection was elevating and clinical symptoms were disappeared was detected by the VIDAS instrument platform verification. There were significant differences between A and B. The detection result of reagent A was [1594.15±2286.18], and reagent B was [912.97±1814.06]. There were significant differences between them (t=3.654, P<0.05). In addition, the consistency of the two reagents was poor (r=0.419, P<0.05). Conclusion:Comparing two reagents to detect D-dimer quantitatively by automated coagulation analyzer ACL-TOP, D-DHS owns greater ability to exclude false-positive results, so does the ability of anti-interference with jaundice, hemolysis and chylemia.

Objective To observe the antioxidant effects of 99Tc-MDP on the brain of aged mice induced by D-galactose.Methods A total of 48 healthy female Kunming mice were divided into 6 groups by random number table method:normal control group,model group,vitamin E (Vit E) group,groups treated with low,middle and high dosages of 99Tc-MDP.Except the normal control group,mice of each group were injected with 10％ D-galactose saline subcutaneously in the neck for 42 d.At the same time,the 99Tc-MDP groups were given different dosages of 99Tc-MDP (1.75× 10-5,3.50× 10-5,0.70× 10-4 μg/g according to the body mass) respectively into abdominal cavity twice a day.Vit E group was given Vit E by intragastric administration from the first day.The model group was injected with saline every day.After behavioral testing,mice serum samples and brain tissue samples were collected for the determination of superoxide dismutase (SOD),malondialdehyde (MDA),glutathione peroxidase (GSH-Px) and monoamine oxidase (MAO)levels.Then the mice were sacrificed and the brain tissue was taken for HE staining.The independent-sample t test was used for data analysis.Results The mice model was established successfully.SOD levels in brain tissue of groups injected with low,middle,high dosages of 99Tc-MDP,Vit E group were (19.06± 6.44),(20.41±4.02),(22.24±3.76),(24.71±5.09) U/mgprot,respectively,all of which were higher than that of the model group((11.32±2.90) U/mgprot;t=3.099 6-6.504 6,P＜0.05 or ＜0.01).There were similar results for GSH-Px (t =2.214 1-4.145 7,P＜0.05 or ＜0.01).MDA and MAO levels in brain tissue of 99Tc-MDP groups were lower than those of the model group (t =2.140 3-3.057 8,all P＜0.05).Compared to normal control group,the hippocampus in model group showed reduced cell number and layers,disordered structure,with part of the cells in smaller volume and abnormal nuclear shape.In Vit E group and the three 99Tc-MDP groups,no significant change of neuron was observed compared with normal control group,the degeneration and necrosis of hippocampal cells were mild compared with model group,the cell number and morphology were normal,and the structures were clear.Conclusion 99Tc-MDP may increase the activities of SOD,GSH-Px and reduce the levels of MDA and MAO in brain tissue of aged mice,thus it may be helpful in delaying the brain aging.

Objective To investigate the association of T help (Th) lymphocyte and heart rate variability (HRV) in congestive heart failure (CHF) patients. Methods Ninety-six patients with CHF and thirty healthy persons were enrolled in the study. Time-domain HRV analysis was performed based on 24 hour Holter Electrocardiogram (ECG) monitoring. Interferon-γ (IFN-γ) was used as markers for the differentiation of Th1 subsets and interleukin-10 (IL-10) for the Th2 subsets. IFN-γand IL-10 in CD4 + T lymphocytes were quantified by 3-color flow cytometry. Results The frequency of IL-10-Producing T Cells in the CHF group was significantly lower than those in the healthy control ( ( 16.4 ± 5.8 ) % vs. ( 26.8 ± 3.7 ) %, t = 9. 243, P ＜ 0. 001 ). The frequency of IFN-γ in the CHF group ( ( 18.4 ± 7.3 )% ) was significantly lower than that in the healthy controls ( (7.3 ±4.6) % ,t =7. 917, P ＜ 0. 001 ). The following index of HRV in the CHF groups were all significantly lower than those in the healthy controls: (98. 6 ± 21.3) ms vs. ( 145. 1 ± 42. 6) ms for SDNN, (83. 9 ± 22.4) ms vs.(136.5 ±39.6)ms for SDANN, (40.6 ± 14.5) ms vs. (55.8 ± 17.9) ms for SDNNI, (20. 7 ± 12.9) ms vs.(29.1 ± 12.6) ms for RMSSD, (5.6 ± 3.7 ) % vs. ( 11.8 ± 4.4) % for PNN50 ( Ps ＜ 0.05 ). In CHF patients, the frequency of IL-10 were positively associated with SDNN, SDANN, SDNNI, RMSSD and PNN50 ( r = 0. 49,0. 57,0. 58,0.47 and 0. 52 ,respectively,Ps ＜ 0.01 ). In the CHF patients, the frequency of IFN-γand IFN-γ/IL-10 ratio were negatively associated with SDNN ,SDANN ,SDNNI, RMSSD and PNN50 ( r = - 0. 49, - 0. 54, - 0. 57, - 0.52,- 0.53, - 0. 52, - 0.64, - 0.57, - 0. 58, - 0. 67, Ps ＜ 0. 01 ). Conclusions Autonomic nervous system is involved in the regulation of the balance of Th1/Th2 in patients with CHF. Sympathetic nerve system enhances the effect of Th1 ,whereas parasympathetic nervous system enhances the effect of Th2.

Objective To investigate current status of learned resourcefulness in pregnant women,and to identify its influencing factors.Methods With convenient sampling,a self-made General Data Questionnaire and the Chinese version of Self-Control Schedule (C-SCS) was used to survey 360 pregnant women in the antenatal clinic in one three-class hospital in Guangzhou.Results The mean score of learned resourcefulness in pregnant women was (115.00±14.42) and its influencing factors included education degree,gravidity,pregnancy school training and planed pregnancy.Conclusions Learned resourcefulness in pregnant women loads upper level.Medical staff should take targeted interventions to strengthen it and facilitate the mental well-being of pregnant women.

A real-time tele-monitoring system for physiological multi-parameter based on Internet is introduced,and physiological signals are transmitted by P2P network technology.The experiment results show that the speed of data transfer has been improved greatly using P2P technology,and physiological signals,such as ECG,can be monitored in real-time.

Objective To analyze the association of Crohn's disease(CD)with vitamin D receptor(VDR) gene polymorphisms. Methods After collecting 326 CD patients and 464 healthy controls,the four single nucleotide polymorphisms of VDR (FokI, BsmI, ApaI and TaqI) were examined by a SNaPshot technique. Results Compared with those in controls,the frequencies of mutant allele(A)and genotype(GA+AA)of BsmI were significantly decreased in CD patients(both P=0.001). The similar conclusions were also drawn for the mutant allele(C)and genotype(TC+CC)of TaqI(both P<0.05). In further stratified analysis,compared with those in controls,the mutant alleles and genotypes of BsmI and TaqI were significantly reduced in stenotic type CD patients (all P<0.0083). The analyses of linkage disequilibrium(LD)and haplotype showed that BsmI,ApaI and TaqI were in a strong LD,and the formed haplotype AAC was significantly lower in CD patients than that in controls (P <0.05). Conclusions VDR(BsmI and TaqI)polymorphisms are significantly related with the reduced susceptibility to CD,especially for patients with stenotic CD. Moreover,the haplotype AAC might engender a reduced risk of CD.

Objective To investigate the association of ulcerative colitis (UC) with fork head/ winged helix transcription factor-3 (Foxp3) polymorphisms in Han population in Zhejiang province,China.Methods A total of 381 UC patients and 490 healthy controls were enrolled in this study.The four single nucleotide polymorphisms (SNPs) of Foxp3 (rs3761547,rs2232365,rs2294021,rs3761548) were examined by SNaPshot.The analyses of linkage disequilibrium (LD) and haplotype were also performed in all study subjects.Results When male and female UC patients were compared with their corresponding controls respectively,the alleles and genotypes of the four SNPs were not statistically different (all P ＞0.05).According to severity and location of the disease,the UC patients were divided into different subgroups.The alleles (C,G,A) of (rs2232365,rs2294021,rs3761548) were more frequent in male patients with severe UC than in the male controls (69.6％ vs 34.3％,P =0.001;69.6％ vs 34.3％,P =0.001;39.1％ vs 14.4％,P =0.002,respectively).As compared with the female controls,the alleles (C,G,A) and genotypes (TC + CC,AG + GG,CA + AA) of (rs2232365,rs2294021,rs3761548) were significantly increased in the female patients with severe UC (51.9％ vs 38.0％,63.5％ vs 39.2％,53.8％ vs21.4％,80.8％ vs57.7％,84.6％ vs58.4％,76.9％ vs34.7％,all P＜0.05).The four SNPs above were shown to be in a strong LD both in male and in female subjects.When male and female UC patients were compared with their corresponding controls respectively,nevertheless,each haplotype frequency was not statistically different (all P ＞ 0.05).Conclusions Foxp3 (rs2232365,rs2294021,rs3761548) variations might engender the increased risk of severe UC in Chinese Han patients.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of TLR4 and its role in lipopolysaccharide (LPS)-induced NF-κB activation and CD40 expression in rat peritoneal mesothelial cells (RPMCs). Methods RPMCs were harvested from Spragne-Dawley rat peritoneal cavity and maintained under defined in vitro condition. The cells were treated with Ang Ⅱ at different concentrations (10-9, 10-8, 10-7, 10-6 mol/L) and exposed to Ang Ⅱ (10-7 mol/L) for different times (1, 2, 4, 8, 12, 24, 48 h for mRNA and 6, 12, 24, 36, 48 h for protein, respectively). Meanwhile, the influence of AT1 receptor antagonist (AT1R, losartan, 10-5 mol/L) and AT2 receptor blocker (AT2R, PD123177, 10-5 mol/L) on the TLR4 induced by Ang Ⅱ was observed. After synchronization for 24 hours, the cells were randomly assigned to four groups: the control group, the Ang Ⅱ (10-7 tool/L) group, the LPS (1 mg/L) group, the Ang Ⅱ (10-7 mol/L) plus LPS (1 mg/L) group, which were used to investigate the effects of Ang Ⅱ on the NF-κB activation and CD40 expression induced by LPS. The mRNA expression of TLR4 and CD40 was measured by RT-PCR and the protein abundance of TLR4, NF-κB p65, phospho-p65, IKBα and phospho-IκBα were analyzed by Western blot. Immunofluorescence was performed to determine the subcellular localization of p65 subunit of NF-κB. Results (1) Treatment of RPMCs with Ang Ⅱ resulted in a concentration-dependent increase in the expression of TLR4. Ang Ⅱ at 10-9, 10-8, 10-7 and 10-6 mol/L increased TLR4 mRNA expression by 70.5%, 89.5%, 102.9%, and 121.9%, respectively and protein expression by 12.1%, 27.7%, 51.2%, and 41.6%, respectively (P<0.01). Treatment of RPMCs with 10-7 mol/L Ang Ⅱ resulted in a time-dependent increase in the expression of TLR4, with the peak of mRNA expression at 8 and 12 h (P<0.01) and the protein expression at 12 and 24 h (P<0.01). (2) Losartan antagonized Ang Ⅱ-stimulated expression of TLR4 by 33.5% (P<0.05), PD123177 had no such effect (P0.05). (3) Treatment of RPMCs with LPS (1 mg/L) for 60 rain significantly increased the ratio of phospho-IκBα to IκBα by 362.6% (P< 0.01) , phospho-p65 to p65 by 67.4% (P<0.05), and LPS (1 mg/L) for 4 h significantly increased the expression of CD40 mRNA by 299.9% (P<0.01) compared to the control group. In comparison to the LPS (1 mg/L) group, preincubation of RPMCs with AngⅡ (10-7 mol/L) for 24 h then treated with LPS (1 mg/L) for 60 rain significantly increased the ratio of phospho-IκBα to IκBα by 49.1% (P<0.01), phospho-p65 to p65 by 29.3%(P<0.05), and LPS (1 mg/L) for 4 h significantly increased the expression of CD40 mRNA by 56.8%(P<0.01). (4) The p65 subunit of NF-κB was dominantly distributed in the cytoplasm in the control and Ang Ⅱ group. Following exposure to LPS for 60 min, p65 subunit labeling was upregulated and translocated into the nuclei. A significantly increased nuclear staining of p65 in ceils treated with Ang Ⅱ plus LPS were observed. Conclusions Ang Ⅱ induces the expression of TLR4 in dose- and time-dependent manner in RPMCs, resulting in enhanced NF-κB signaling and induction of CD40 expression, Locally produced Ang Ⅱ in the peritoneum may play an amplified role in LPS-induced peritoneal inflammation.

Objective To investigate the expression of CD40 and intercellular adhesion molecule-1 (ICAM-1) treated with lipopolysaccharide (LPS) in rat peritoneal mesothelial cells(RPMC) and the role of NF-κB signal transduction pathway. Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were exposed respectively to different concentrations of LPS for 12 h or treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of CD40 and ICAM-1, the RPMCs were treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of NF-κB and p-NF-κB protein, the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μmol/L or 1 μmol/L ) for 3 h, then treated with LPS for another 3 h, respectively. Expression of CD40 and ICAM-1 mRNA was examined by RT-PCR. Expression of NF-κB and p-NF-κB protein was detected by Western blot. Results Compared with medium control group, stimulation of RPMCs with 1 μg/ml and 5 μg/ml of LPS resulted in a significant increase in the expression of CD40 and ICAM-1 mRNA(P<0.05). 10 μg/ml of LPS had strongest effect on CD40 and ICAM-1 expression compared with that of 1 μg/ml and 5 μg/ml of LPS. Treatment with 5 μg/ml of LPS resulted in time-dependent increase in the gene level of CD40 and ICAM-1, with the peak at 3 h. However, after that time point, the gene level of them was gradually attenuated. Following treatment with LPS (5 μg/ml), the level of p-NF-κB began to increase at 15 min, gradually reached the peak at 1 h, and then decreased. But the level of p-NF-κB at 2 h was still significantly higher than that of medium control. 5 μmol/L of BAY11-7085 decreased significantly the up-regulation of CD40 and ICAM-1 induced by LPS. Conclusion LPS enhanced the expression of CD40 and ICAM-1 on RPMCs in a concentration-dependent and a time-dependent manner. LPS induced expression of CD40 and ICAM-1 depend on the NF-κB signal transduction pathway.

Objective To explore the association of genetic polymorphism of fucosyltransferase (FUT) 2 and FUT3 and expression of Lewis antigen with ulcerative colitis (UC) in Chinese Zhejiang Han population.Methods We genotyped FUT2 (rs281377, rs1047781 and rs601338) and FUT3 (rs28362459, rs3745635 and rs3894326) in 485 UC patients and 580 healthy controls using SNaPshot. By immunohistochemistry method, we also evaluated expression of Lewis a and b antigens in the sigmoid colon of 10 UC patients and 10 patients with benign colonic polyps.Results The frequencies of mutant allele (A) and genotype (GA+AA) in FUT3 rs3745635 were higher in UC patients than in controls (P=0.016, 95%CI 1.339-1.699;P=0.038, 95%CI 1.330-1.742, respectively). Stratified analyses revealed that the frequencies of mutant allele (G) and genotype (TG+GG) of FUT3 rs28362459 were significantly lower in patients with extensive colitis than in those with distal colitis (P=0.001, 95%CI 0.567-0.786;P<0.001, 95%CI 0.503-0.742, respectively). Similar conclusions were drawn for the mutant allele (A) and genotype (GA+AA) of FUT3 rs3745635 in patients with extensive colitis compared to those with distal colitis (P=0.011, 95%CI 0.621-0.900;P=0.006, 95%CI 0.553-0.845, respectively). Although expression of Lewis b antigen in the sigmoid colon did not differ between UC patients and controls, Lewis a antigen expression was higher in the crypt epithelium of both inflammatory and non-inflammatory sigmoid colon of UC patients than in controls (P=0.028).Conclusion Polymorphisms of FUT3 and expression level of Lewis a antigen might be associated with UC.