Aim To investigate the transcytosis mechanism of chlorogenic acid(CGA)by using Caco-2 and MDCK(Madin Darby canine kidney) monolayers models.Method ① Caco-2 and MDCK cell models:Caco-2 cell(105 cells/cm2) and MDCK cell(5?104 cells/cm2) were inoculated in Millicell-CM culture plate inserts,and the TEER of cell monolayer were detected to make sure the models are available for experiments.② Permeating experiments: to measure the value of OD of CGA and calculate the cumulative amount.Result CGA could be Absorbed and secreted on two monolayer models.Verapamil could inhibit the secretion at lower concentration of CGA on MDCK monolayer model.P-pg could partly act on the secretion of CGA on Caco-2 and MDCK cell models.Conclusion CGA can secrete and Absorb at the same time across Caco-2 and MDCK cell monolayers,P-pg partly involving in the secretion of CGA.

Horseradish peroxidase (HRP) has been curently used for tracing the interrelationship between neurons in peripheral and central nervnos system. In order to differentiate endo-and exogenous peroxidases, 15 adult, healthy rats were selected for investigating the distribution of endogenous peroxidase in tissues. Fresh cryostat frozen sections of brain were fixed in fixatives of various concentrations and PH, and sections were made after perfusion of the fixative through heart. They were incubated in media containing DAB and hydrogenperoxide. Potassium cynide or sodium azide was used as inhibitators.The reaction of peroxidase in medula oblongata, pons, midbrain, thalamus, hypothamus, hippocampus and striatum etc were studied with dark field and transmitted light microscope. In the superficial layer of the Ⅲ and Ⅳ ventricles and periaqueductal gray substance of midbrain, there were large number of dark brown peroxidase granules with non neuronal distribution.The peroxidase granules were found in the entire or the dorasal part of locus ceruleus, the ventral part of cerebellum and the dorasal part of hippocampus. Small amount of granules were located in medial nucleus of habenularis, stria medullaris, nucleus supraopticus hypothalami and suprachiasmaticus. Large amount of positive reactive granules were found in nucleus arcuate. Some of them were localized in the perineurons of polydendritic neurons but most of them were of nonneuronal distribution. Sparse granules were also observed in the dorsomedial part of nucleus caudatus putamen and dorsal part of nucleus septicus lateralis. In commisure fornix and corpus callosum, the granules were distributed along the nerve fiber bundle. There were large amount of positive reactive granules around the subfornix organ especially on its dorsal part. To sum up, the main distribution of them are periventricular, periaqueductal gray substance and periventricular organs (such as area postrema, locus coeruleus, nucleus arcuata, subfonix organ and several neurosecretory nuclei of hypothalamus). It seems that the distribution of this enzymes might be closely related to synthesis, metabolism and release of the neurosecretion or transmitters of these nuclei and some of them might act through the cerebro-spinal fluid.

Objective:To study the effect and clinical significance of tumor necrosis factor-?(TNF-?) in viral myocarditis(VMC).Methods:Animal model was established by serum TNF-? in VMC mice was assayed by the way of ELISA.In addition,VMC mice were treated by TNF-? mAb and rTNF-?,death rate andmyocardial change of intervention group were compared with control group.Results:Serum TNF-? in VMC mice increased apparently.Inflammatory cell infiltration and myocardial necrosis had ralations with TNF-? in VMC.Inflammatory cell infiltration and myocardial necrosis in the mice of exogenous TNF-? intervention group were more serious than simple VMC group apparently,death rate of mice was also higher than simple VMC group.Inflammatory cell infiltration and myocardial necrosis in the mice of TNF-? mAb intervention group were milder than simple VMC group apparently,death rate of mice was also lower.Conclusion:TNF-? take part in the immunoreaction and pathogenesis of VMC,it has important significance to the therapy of VMC.

Objective To study ultrastructure changes of myocardium and morphic change of cardiomyocyte apoptosis in mice with viral myocarditis(VMC).Methods Our experiment established animal model of VMC by the way of coxsackie virus B3(CVB3) inoculation,then we studied the changes and apoptosis of cardiomyocyte by ways of microscope and electron microscope.Results The myocardial changes and inflammatory cell infiltration were found by ways of microscope and electron microscope from 5 days after virus inoculation in experimental mice,the peak changes were at 7-9 days,and were almost recoverd at 35 days.Apoptotic changes and apoptotic bodies were found by eletron microscope at 7-9 days after virus inoculation in mice with VMC.Conclusions VMC can be caused in mice after CVB3 inoculation,abnormal cardiomyocyte apoptosis can be found in VMC.

Objective To investigate the effects of phenylephrine and norepinephrine on tissue oxygen metabolism in elderly patients undergoing fluid restriction. Methods Eighty elderly patients who had underwent elective colorectal surgery were selected, with ASA of Ⅰ-Ⅱ grade. The patients were divided into restricted fluid administration combined with phenylephrine group (group Ⅰ) and restricted fluid administration combined with norepinephrine group (groupⅡ) by random digits table with 40 cases each. The compound recipe sodium lactate was given at a rate of 5 ml/(kg · h). Small dose of phenylephrine was infused intravenously at 0.1-0.3μg/(kg · min) simultaneously during the surgery in group Ⅰ, or norepinephrine 0.01-0.03 μg/(kg · min) in group Ⅱ. Mean arterial pressure (MAP) was maintained ≥ 65 mmHg (1 mmHg = 0.133 kPa). The intraoperative blood loss was replaced with the equal volume of 6% hydroxyethyl starch 130/0.4. The blood gas analysis, hemoglobin and so on were determined before the surgery (T0), 1 h after beginning of surgery (T1), 2 h after beginning of surgery (T2) and at the time of leaving operation room (T3). The complications were recorded. Results There were no statistical differences in MAP, central venous pressure (CVP), heart rate, lactic acid, arterial partial pressure of oxygen, central venous oxygen pressure, central venous blood oxygen saturation, oxygen uptake rate, hemoglobin, albumin, blood urea nitrogen, creatinine and incidence of complication between 2 groups (P>0.05). The T1-3 hemoglobin and arterial oxygen content (CaO2), T1 and T3 venous oxygen content (CcvO2) and T2 atrerial venous oxygen content differences (Da-cvO2) in groupⅠwere significantly higher than those in groupⅡ, hemoglobin:(103.8 ± 7.9) g/L vs. (110.8 ± 8.6) g/L, (101.7 ± 7.7) g/L vs. (107.3 ± 7.8) g/L and (101.6 ± 6.9) g/L vs. (106.8 ± 5.6) g/L; CaO2: (140.6 ± 9.7) mmol/L vs. (149.6 ± 10.5) mmol/L, (137.5 ± 9.3) mmol/L vs. (144.9 ± 10.2) mmol/L and (137.2 ± 9.2) mmol/L vs. (143.1 ± 8.9) mmol/L;CcvO2:(119.8 ± 10.2) mmol/L vs. (126.4 ± 10.8) mmol/L and (102.3 ± 8.6) mmol/L vs. (108.5 ± 8.9) mmol/L;Da-cvO2:(19.6 ± 3.9) mmol/L vs. (23.0 ± 4.5) mmol/L, and there were statistical differences (P<0.05). Conclusions Phenylephrine has more effect on tissue oxygen metabolism in the elderly patients undergoing fluid-restricted abdominal surgery.

Objective To provide a reference for prevention of trace elements deficiency in children by investigating the distribution and variation law of Ca,Cu,Zn,Fe,Mg of children in Beijing area.Methods Five kinds of trace elements( Ca,Cu,Zn,Fe,Mg) in peripheral blood of 7972 children of Beijing Children's Hospital for health examination from 2010 to 2011 were detected by atomic absorption spectrometer.The children were Beijing urban residents.They were divided into five age groups:infancy group ( 1 - 12 months,1506 cases),toddler age group ( 1 - 2 years,2766 cases),preschool age group ( 3 - 6 years,1918 cases),school age group (7 - 11 years,1576 cases) and adolescence group ( 12 - 18 years,206 cases).According to gender,each group was sub-divided into male and female group.The distributions of the 5 kinds of trace elements were analyzed among different age and gender groups.Results With the age increasing,the trend of the levels of Ca and Cu were downward,but the levels of Zn and Fe were upward,and the level of Mg was stable.For boys and girls,the levels of Cu in infancy group were (21.90 ±2.89) and(21.25 ±2.80) μmoL/L; in toddler age group,they were (21.76 ±2.78)and (21.29 ±2.69) μmol/L; in preschool age group,they were ( 21.32 ± 2.83 ) and ( 20.88 ± 2.84 ) μ mol/L; and in school age group,they were (20.81 ± 3.02 ) and ( 20.36 ± 3.37 ) μmol/L,respectively.The distribution of Cu showed significantly different among different gender groups ( t =4.640,4.475,3.290,2.894; P ＜ 0.01 ).In adolescence group,the levels of Cu were ( 19.53 ± 2.91 ) and (20.30 ± 2.90) μmol/L for boys and girls respectively and there was no significant difference ( t =- 1.796,P ＞ 0.05 ).In each gender group,the content of Ca,Zn,Fe,and Mg was not significantly different between boys and girls.It was common for the deficiency of Zn and Fe in all ages.Their total deficiency rates were 58.9％ and 19.2％.The deficiency rates of Zn and Fe in infancy were 67.7％ and 42.3％.Conclusions The distributions of Ca,Cu,and Mg in peripheral blood of children in Beijing become reasonable.Fe and Zn are deficient in different degree,and should be rationally used according to clinical guide.

Objective To investigate whether the expression frequency and function of peripheral and liver NK cells was correlated with the liver injury in patients with hepatitis B virus related acute-on-chronic liver failure(HBV-ACLF).Methods Peripheral blood samples were obtained from fifteen HBV-ACLF patients and fifteen chronic hepatitis B patients.The frequency of peripheral NK cells (CD3-CD56+) and CD107a expression on surface of peripheral NK cells were detected by multicolor flow cytometry.Expression of IFN-γ by peripheral NK cells was detected by intracellular cytokine staining.Needle biopsy liver tissues were obtained from twenty patients with HBV-ACLF,ten patients with mild CHB,and expression of live NK cells (CD3-CD57+) was analyzed by dual immmunohistochemical staining of CD 3 and CD57.Results There was no significant difference in the frequency of peripheral NK cell and IFN-γ expression by peripheral NK cells between HBV-ACLF and mild CHB patients.However,CD107a expression on surface of peripheral NK cells of HBV-ACLF patients was remarkably higher than that of CHB patients.The frequency of liver CD57+NK cell of HBV-ACLF patients was remarkably higher than that of CHB patients(95.1 ±21.3/low power field vs 9.5±10.6/low power field,P＜0.01).Further analysis revealed that the frequency of liver CD57+ NK cells in HBV-ACLF patients was positively correlated with the TBIL level.Conclusion The enhanced cytotoxic activity of peripheral NK cells and the recruitment of liver CD57+ NK cells may aggravate immune-mediated liver injury and promote the apoptosis and necrosis of hepatocytes.

Objective To investigate the role of the chemokine receptor CXCR3 and its ligands in the migration of lymphocytes and acute hepatic failure. Methods BALB/cJ mice (6-8 weeks, female) were intraperitoneally injected with 100 PFU mouse hepatitis virus-3(MHV-3). The proportions and numbers of T cells and NK cells in liver, spleen, and blood as well as the expression of CXCR3 in T cells, and NK cells post MHV-3 infection was analyzed by flow cytometry. The hepatic mRNA level of the CXCR3-associated chemokines(CXCL9 and CXCL10) was detected by real-time PCR. A transwell migration assay was used to assess the chemotactic effect of MHV-3-infected hepatocytes and CXCL10 on the splenic lymphocytes. Results Following MHV-3 infection, the number of hepatic NK cells and T cells and the frequencies of hepatic NK cells and T cells expressing CXCR3 increased markedly; however, in the spleen and peripheral blood, they both decreased significantly. Moreover, the hepatic mRNAs levels of CXCL9 and CXCL10 were significantly elevated post infection. The transwell migration assay demonstrated that MHV-3-infected hepatocytes have the capacity to attract and recruit the splenic NK cells and T cells, and CXCL10 plays a key role in lymphocyte mobilization from the spleen. Conclusion Interactions between CXCR3 and its ligands (CXCL9 and CXCL10),especially CXCL10 may play a key role in the recruitment of intrahepatic lymphocytes and subsequent necroinflammation and acute hepatic failure in MHV-3 infection.

Objective:To study the correlation of elastase(EA) and ? 1-antitrypsin (? 1-AT) level in gingival crevicular fluid(GCF) of different periodontal status and their roles in periodontal inflammatary pathogenesis. Methods: 62 volunteers aged 45～51 years old were enroled.Their periodental status were examined and grouped into healthy periodontium (H),8 cases,marginal gingivitis (MG),12 cases,mild chronic periodontitis (MCP),20 cases and advanced chronic periodontitis (ACP),22 cases.EA in GCF were measured with a chromogenic low molecular weight substrate reaction and the ? 1-AT with ELISA. Results: Significantly positive correlation was found between GCF-EA activity and clinical periodontal parameters (P

Objective To explore the application of bacterial community diversity profiling in forensic comparison of soil.Methods The DNA profilings of bacterial community from 5 soil samples of different source and 4 samples of same source were obtained through the specific amplification of 16S rDNA segments followed by terminal restriction fragment length polymorphism(T-RFLP)analysis,and the similarity indexes between different soil samples were calculated.Result The ranges of similarity indexes between different sources of soil samples were 0.3～0.44,and the ranges between same sources of soil samples were 0.76～0.87.Conclusion There is a great relativity between the bacterial community diversity profiling and the source of soil sample.