Objective To detect the expression of type Ⅰ interferon in monocyte-derived dendritic cells(MoDCs)after Toll like receptor(TLR)3 triggered in patients with chronic hepatitis B(CHB),and to evaluate immune responses of CHB patients and its roles in the mechanisms of persistent infection of hepatitis B virus(HBV)and chronicity of hepatitis.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and purified using magnetic beads(plasma was saved simultaneously)from 26 CHB patients and 18 healthy volunteers(HV).Dendritic cells(DCs)were induced and proliferated in a culture medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin(rhIL-4).EX3s were stimulated with Poly Ⅰ:C and the supernatants were collected at 0 h and 24 h after stimulation.Type Ⅰ interferon(IFN-α and IFN-β)in plasma and supernatants were examined by enzyme linked immunosorbent assay (ELISA).Results The levels of type Ⅰ interferon in plasma were not significantly different in groups of HV and CH B.IFN-α and IFN-β expressions in supernatants before Poly Ⅰ:C stimulation were(80.00±16.15)ng/L,(36.39±13.90)ng/L in CHB group and(76.76±15.90)ng/L,(37.14±13.68)ng/L in HV group,respectively.And there were no statistical differences between two groups(t=1.651,t=0.178;both P＞0.05).IFN-α expressions in supernatants at 24 h after stimulation in two groups were both higher than those before stimulation(at 0 h),but there were no statistical differences(t=1.534,t=1.243;both P＞0.05).IFN-β expressions in supernatants at 24 h after stimulation in HV group was(54.57±16.80)ng/L,which was significantly higher than that at 0 h(37.14±13.68)ng/L(t=4.061,P＜0.05).However,there was no significant difference at 24 h than tht at 0 h in CHB group(t=1.796,P＞0.05).At 24 h after stimulation.IFN-β level was(54.57±16.80)ng/L in HV group,which was significantly higher than that[(41.64±12.57)ng/L]in CHB group(t=2.921,P＜0.05).Conclusions Functions of MoDCs from CHB patients are impaired and MoDCs could not express type Ⅰ interferon normally.Expression of type Ⅰ interferon after TLR3 triggered in CHB patients is mainly IFN-β.

Objective To elucidate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) in patients with chronic hepatitis B (CHB), and to explore the correlation between hepatitis B virus (HBV) persistent infection and TLR3 expression. Methods Sixty CHB patients (CHB group) and 20 healthy controls (control group) were enrolled. The peripheral blood mononuclear cells (PBMCs) were isolated and CD14~+ monocytes were sorted by immunomagnetic beads. Immature DCs (imDC) were induced and proliferated in vitro and mature DCs (mDC) were obtained after the poly I:C stimulation. The expression of intracellular TLR3 mRNA was detected by real-time polymerase chain reaction (PCR), and surface markers [CD80 and human leucocyte antigen (HLA)-DR] were determined by flow cytometry after 48 h of stimulation. The comparison of quantitative data was done using t test. The qualitative data were compared using chi-square test.Results The mean fluorescence intensities (MFI) of intracellular TLR3 of imDC before poly I:C stimulation in CHB group and control group were 1212.05 ± 250.80 and 1192.95 ± 301.40,respectively, which were not significantly different (t = 0. 280, P>0. 05). While after stimulation,those were 1352.98± 313.67 and 1593. 00± 349. 65, respectively, the latter was significantly higher than the former (t = 2. 880, P<0. 05). The levels of TLR3 mRNA inside mDCs in both groups were increased after poly I:C stimulation, which were 0. 1204 ±0.0267 and 0. 1780 ± 0.0664, respectively in CHB group and control group, and that in control group was significantly higher (t = 3. 909, P<0.05). Furtherly, patients in CHB group were divided into HBeAg(+ ) and HBeAg( -) subgroups.After stimulation, the MFI and mRNA of TLR3 inside mDC were greatly elevated in both subgroups,but there were no difference between these two subgroups (t = 0. 366, P>0. 05). Conclusions The intracellular expressions of TLR3 in mDC in CHB group and control group are obviously increased after the poly I:C stimulation, but the increased level in CHB group is lower than that in control group. The results suggest that the insufficiency of TLR3 synthesis may be related to the HBVpersistent infection.

Objective To evaluate the effect of human bone morphogenetic protein 2 (hBMP-2)/Bone Mesenchymal Stem Cells (BMSCs)/demineralized bone matrix(DBM) on repairing rabbits’femoral head after necrosis and to explore the new treatments for femoral head necrosis. Methods Femoral head necrosis models was established by clinical core decom?pression combined with liquid nitrogen frozen. Then, animals were randomly devided into 4 groups (n=12 per group):Group A were not implanted anything as control group, Group B were implanted with DBM. Group C were implanted with hBMP-2/DBM. Group D were implanted with hBMP-2/BMSCs/DBM. Four rabbits from each group were sacrificed at 4,8 and 12 weeks after surgery to evaluate the the repairing effect of Osteonecrosis of the femoral head (ONFH) through X-ray examina?tion, observation of the specimen and HE staining. Results X-ray revealed defect of femoral head in Group A without clear bone formation. There is a little fibrous hyperplasia and no obvious osteogenic response. By contrast, the femoral head defect areas became fuzzy in group B, group C and group D with new bone trabeculars. And the regenerate phenomenons of group D were significantly better than that of group B and group C of the same time point. As to the Lane-Sandhu X Ray scores, it is lower in group A than that in group B;It is lower in group C than that in group D(P<0.05). There is no statistical difference between Group B and Group C. General observation of the specimen revealed that the femoral head of group A collapsed with drilling holes. The femoral heads of group B and group C showed no collapse but the drilling holes existed. Femoral head in group D was not collapsed and the drilling holes disappeared. HE staining showed that bone trabeculars became ne?crotic and fragmented in Group A with a lot of air trapped cells. There were newborn immature bone trabeculars and osteo? blasts in group B and group C. Group D were of large number of bone cells, fat cells, and newborn mature bone trabeculars. The ratio of empty lacuna is higher in Group A than that in Group B;it is higher in Group C than that in Group D(P<0.05). Conclusion hBMP-2/BMSCs/DBM can induce BMSCs differentiation into osteoblasts after being implanted. It has good re?pairing effect on ONFH with good application prospect.

Objective The use of in vivo cryotechnique (IVCT) in combination with electrocardiograph (ECG) to study cardiac microcirculation under different hemodynamic conditions in living mouse.Methods Living mouse heart monitored by electrocardiograph was suffered from IVCT and freezing substitution under normal blood flow,myocardial ischemia or cardiac arrest conditions.Hematoxylin eosin (HE)staining,Schiff's staining and immunofluorescence staining for serum albumin,immunoglobulin were utilized on continuous paraffin sections,respectively.Confocal microscopy and statistical analyses were used.Results Comparing with normal hemodynamics,microvascular red cell volume reduction,morphology changed,myocardial cell glycogen loss,serum albumin ectopic distribution to myocardial cytoplasm,T tubular network failure and spacing width were happened in myocardial iscbemia condition; different shapes of red blood cells,myocardial cells glycogen deficiency,T tubular network failure and interval narrowing were found under cardiac arrest conditions.Conclusions Cardiac microcirculation,pathological changes of myocyte and its surrounding microenviroument in living mouse heart can be immediately captured in situ by the application of IVCT and ECG.

Objective To set up a new method for fast determining and identifying Xintong Oral Liquid by acousto-optic tunable filter-near infrared(AOTF-NIR)spectroscopy.Methods Identifying method by NIR spectroscopy combined with principal component analysis(PCA),and determining model of qualitative and quantitative analyses based on PLS1 algorithm.Results The XT-C identifying model could be used to identify Xintong Oral Liquid,the determining model of qualitative and quantitative analyses with better accuracy,RMSEP of the models for puerarin was 0.137 1,the determination coefficients was R2=0.984 5.The correlation coefficient of the true value and predication value from validation was r2=0.996 4.The average recovery of the predication set was 101.9%.RSD for precision was 1.98%,RSD for stability was 1.57%.Conclusion The method is a quick and simple assay technique with low cost,which is able to be used in the qualitative and quantitive analyses on Xintong Oral Liquid.

Objective To analyze levofloxacin resistance in Helicobacter pylori (Hp) and the sequence difference of gyrA gene in levofloxacin resistance and sensitive Hp strains. To explore the function of gyrA gene mutation in the development of levofloxacin resistance Hp strain.Methods From July 2007 to December 2008 in Department of Gastroenterology, Jinhua People hospital of Zhejiang Province, the gastric mucosa from gastroscopy biopsy of chronic gastritis and peptic ulcer patients were cultured in Hp selective medium under microaerobic condition at 37 degrees for three to five days. The Hp strains were isolated and identified by oxidase test, catalase test,urease test and UreA gene detecting. The levofloxacin susceptibility was determined by E-test and then resistance and sensitive strains were screened. The genomic DNA of Hp strains was isolated. The gyrA gene was amplified by PCR and the sequences were analyzed. Results 38 clinical isolated Hp strains were passed the levofloxacin susceptibility E-test, among those the minimum inhibitory concentration (MIC) of 12 strains was over 1.0 μg/ml, the percentage of resistant strain was 31.58%, while the sensitive stains was 68.42%. The gyrA gene sequence result indicated 10 resistant strain with 261, 271 and 272 10 site mutation, 2 strains with C261A mutation, one strain with C261G mutation, two strains with G271A mutation, 2 strains with A272G mutation, 2 strains with C261A,G271T andA272G mutation, 1 strain with C261G and A272G mutation. However, no mutation sites were found in 26 sensitive strains. Conclusion The rate of levofloxacir-resistance in isolated clinical Hp strain was high. The drug-resistance was associated with 261,271 and 272. site mutation of gyrA gene.

Objective To investigate the molecular mechanisms for the development of cardiac hypertrophy in hypertension,the present study provided the differential protein expression analysis of hypertrophied heart at differ- ent stages in spontaneously hypertensive rats (SHR).Methods The profiles of protein expression of left ventricu lar myocardium in SHR and its normotensive control Wistar-Kyoto (WKY) rats at the age of 4 and 20 weeks were analyzed with two-dimensional gel electrophoresis (2-DE) in combination with matrix assisted laser desorption ioniza- tion-time of flight (MALDI-TOF-TOF) mass spectrometry.Results Although the blood pressure of SHR was normal at 4 weeks age,hypertrophy of the left ventricle had already developed.The expression pattern in the hy- pertrophic myocardium was found 27 modulated proteins,20 of which were identified.These proteins are involved in reactions of energy metabolism,mitoehondrial oxidative phosphorylation and oxidative stress,etc.The expres- sion of 13 proteins was significantly changed in SHR rats at early stage prior to the development of sustained hyper- tension,while the expression changes of other 7 proteins occurred only at late stage in SHR rats when the blood pressure was significantly elevated.Conclusions lncrease in glycolysis and decrease in oxidation of fatty acid and glucose was shown in the hypertrophied myocardium from early stage in SHR prior to the development of hyperten sion.The significant changes in protein expression of the mitochondrial electron transport chain and antioxidative molecules support the hypothesis that oxidative stress promotes and accelerates the development of hypertensive car- diac hypertrophy.

Objective To study the feasibility of DCE-MRI application in the orthotopic transplantation model of gastric cancer in nude mice.Methods Orthotopic transplantation model of gastric cancer in nude mice was established, and 15 models underwent DCE-MRI using the contrast agent of gadopentetate.Subsequently, the tumor was dissected in order to detect the MVD.The MVD was compared between orthotopic transplantation tumor model of gastric cancer and normal gastric mucosa.Results Fifteen nude mice with orthotopic transplantation of gastric cancer successfully underwent DCE-MRI examination.As for the cancer, the values of Ktrans, Kep and Ve were (2.11±0.44) min-1, (4.59±0.93) min-1, and 0.46±0.06, respectively.The MVD in gastric cancer tissues was significantly higher than that in normal gastric mucosa (χ2=16.205, P<0.001).Conclusion DCE-MRI can be used for noninvasively quantitative evaluation of vascular parameters of gastric carcinoma.

Objective To study the clinical values of positron emission tomography(PET)in pre-term and term newborn infants through observing neonatal brain by 18F-fluorodeoxyglucose(18F-FDG)PET.Methods The brain by 18F-FDG PET in 11 term and 7 pre-term newborn infants after administration of 0.1 mCi /kg 18F-FDG were observed.There were 11 males and 7 females,who were normal by brain computed tomography or magnetic resonance imaging.Results The brain 18F-FDG PET image in pre-term and term newborn infants was relatively high in thalamus,and relatively low in cerebral cortex,whereas the total brain was different with adults.Especially in the area of cerebral cortex,the uptake of glucose was relatively higher,and the structure of brain 18F-FDG image was more clear in term infants than that in pre-term infants.Conclusion Neonatal brain picture by 18F-FDG PET is a new tool for predicting the brain function,and its clinical values need further investigating.

Objective To explore the changes of positron emission tomography(PET)in newborn infants with HIE through 18F-fluorodeoxyglucose(18F-FDG)and it's significance.Methods Eleven healthy newborn infants and 8 newborn infants with HIE were selected.Among the healthy newborns,7 cases were male and 4 cases were female,and the mean birth-weight was(3 350?620)g,the gestational age was(37.9?1.3)weeks.Among the HIE neonates,5 cases were male and 3 cases were female,and the mean birth weight was(3 180?390)g,the gestational age was(37.1?2.4)weeks.There were no significant differences of sex and gestational age between the 2 groups.The examination time was form 3 to 21 d,and the mean age was(8.7?3.9)d.PET of the children in 2 groups were observed after 0.1 mCi/kg 18F-FDG injected 30 min.Results The brain 18F-FDG PET image in newborn infants was relatively high in thalamus,and relatively low in cerebral cortex,whereas the total brain was different with that of the adults,and that was not as clear as that of adults.Especially in the area of cerebral cortex,the uptake of glucose was relatively higher.The structure of brain 18F-FDG image was significantly changed in newborn infants with HIE,especially increased in the areas of peripheral ventricle and hypophloeodal cerebral white matter,and there was a remarkably bilateral asymmetry.Conclusions Neonatal brain picture by 18F-FDG PET is a new tool for predicting the brain function,and its clinical values need further investigating.