ObjectiveTo study the impact of quercetin on LPS-stimulated macrophage inflammatory cytokine release.MethodsDetermined by MTT assay with different concentrations of quercetin on the activity of RAW 264.7 cells.Cells were divided into five groups:A (control group),B group (LPS intervention group),C (LPS+quercetin 50 μmol/L),D group (LPS+quercetin 150 μmol/L) and E group (LPS+quercetin 250 μmol/L).Cells expression of toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (Myd88) and NF-kappa B (p65) mRNA in each group was determined by real-time determination; Supematant IL-6 and TNF-α secretion level was tested by ELISA determination.ResultsThe quercetin at 0 to 250 μmol/L concentration range on the activity of RAW 264.7 cells did not significantly affect.Compared with the control group,quercetin can effectively reduce the level of LPS-induced TLR4 and of MyD88 and of NF-κB (p65) of rmRNA expression and supematant IL-6 and TNF-α secretion,and dose-dependent manner(P＜0.01).ConclusionQuercetin may reduce macrophage inflammatory cytokine secretion by inhibiting the activation of TLR-Myd88-NF-κB pathway.

Objective To study the effect of candesartan (Can) on the LPS‐induced proinflammatory cytokines release in VSMCs ,and to explore the role of TLR4‐mediated signaling pathway in this process .Methods Rat VSMCs were primarily cul‐tured ,and the effect of different concentrations of Can on VSMCs activity was observed by M TT assay .The cells were divided into 5 groups :A(control) ,B(LPS intervention) ,C(LPS + 10 - 7 mol/L Can) ,D(LPS + 10 - 7 mol/L Can ) and E (LPS + Can 10 - 5 mol/L Can) .mRNA and protein levels of Toll‐like receptor‐4(TLR4) ,myeloid differentiation primary response gene 88(Myd88)and nu‐clear factor‐κB(NF‐κB) nuclear translocation were determined by real‐time PCR and Western blot ,respectively ;IL‐1β and TNF‐αconcentration were detected by ELISA .The production of intercellular reactive oxygen species(iROS) was measured by the DCFH‐DA assay .Pretreating VSMCs with the inhibitors against TLR4 ,NADPH oxidase ,and NF‐κB ,or a combination with candesartan and these inhibitors ,and then the expression of IL‐1β and TNF‐α was measured by ELISA .Results Can in the concentration range of 10 - 8 - 10 - 3 mol/L had no significant effect onVSMCs activity .Compared with th control group ,Can could effectively inhibit LPS‐induced VSMCs ,IL‐1β and TNF‐α release ,decreased the mRNA and protein expression of TLR4 ,Myd88 ,reduced the produc‐tion of iROS ,inhibited the NF‐κB(p65) nuclear translocation with a concentration‐dependent manner(P< 0 .05) .Anti‐TLR4 anti‐body ,DPI ,and PDTC all inhibited LPS‐induced inflammatory cytokines release in VSMCs ,and the combination of Can and these blockers showed stronger anti‐inflammatory effect .Conclusion Can can decrease the releaseof inflammatory cytokines IL‐1β and TNF‐α in VSMCs stimulated by LPS ,which is realized by inhibiting the signaling pathway of TLR4/Myd88‐iROS‐NF‐κB .

Objective　To explore the clinical superiority of two kinds of chopping nucleus methods in phacoemulsification——etch and turn technique, stop and chop technique.Methods　Three hundred and sixty-five eyes were performed with phacoemulsification, among which 85 eyes were performed with turn technique and 280 eyes were performed with chop technique.The results of the two kinds of chopping nucleus were analyzed.Results　(1)Time of chopping nucleus:time of turn technique averaged 81.26 seconds while chop technique averaged 40.71 seconds; (2)Visual acuity in the first week after operation:turn technique:vision of 76 of 85 eyes were 0.5 of better, 7 eyes were 0.2～0.5(8.2%) and 2 eyes were 0.2 or worse; chop technique: vision of 253 of 280 eyes were 0.5 or better (90.4%), 21 eyes were 0.2～0.5(7.5%) and 6 eyes were 0.2 or worse (2.1%);(3)Reaction of corneal endothelial cells after the operation: central endothelium rugosity appeared in 4 eyes (4.7%) with turn techinque and in 7 eyes (2.5%) with chop technique. Conclusion　The operation of the former is less difficult and the chopping nucleus time is longer. The time of the operation on the latter is shorter and it is appriate for hard nucleus and should be applied and spreaded. There are no marked differences as for the vision after the operation and the microcorneal injury between them.

Heparin and low molecular weight heparin have been widely used in clinical therapy as anticoagulants in cardiovascular disease and in hemodialysis. Crude heparin is usually prepared from porcine intestinal mucosa. Purified heparin is a mixture of polysaccharides consisting mainly of repeating GlcNS(6S)-IdoA2S disaccharides and other disaccharides with different GlcNAc/GlcNS±3S±6S-GlcA/IdoA±2S residues. Heparin injections are drugs prepared from heparin active pharmaceutical ingredient ( API ) that is prepared from crude heparin. Low molecular weight heparins are dominant heparin-based drugs used clinically, which are prepared by degrading heparin into smaller sizes. As a result, low molecular weight heparins are sharing the same major disaccharides but have different reducing and non-reducing ends. In current study, we focused on the disaccharide compositional analysis of clinically used heparin and heparin-based drugs. HeparinaseⅠ,II, and Ⅲ were used to degrade all heparin and heparin-based drugs including heparin sodium injection, Enoxaparin sodium injection, Nadroparin calcium injection, Dalteparin sodium injection, Fondaparinux sodium into disaccharides. All the degraded products were analyzed by strong anion high perforance liquid chromatography ( SAX-HPLC) coupled with an UV-detector. Commercially available unsaturated disaccharide standards were then used for structral identification. Furthermore, unusual disaccharides present in Nadroparin, Dalteparin, and Fondaparinux were confirmed by reversed-phase ion pair HPLC coupled with mass spectrometry. The developed method produced detailed structural information, which should be useful for quality control of heparin and heparin-based drugs.

Objective To investigate the clinical significance of detecting mammaglobin in sentinel lymph nodes(SLN) of breast cancer. Methods In 32 breast cancer patients, methylene blue was injected around parechyma of breast cancer to locate the SLNs, and nested RT PCR(reverse transcriptase polgmecane chain reaction) was used to examine the expression of mammaglobin mRNA. Results The SLN was successfully identified in 30 of the 32 cases(93.8%).The micrometastases detection between group SLNs and non SLNs had significant statistical difference(P

Objective To construct a recombinant Ientiviral vector of mFVII/Fc and investigate its transfective efficiency into human bone mesenchymal stem cells (hBMSCs),and to detect the expression of mFVII/Fc fusion gene in vitro. Methods Coagulation factor VII (FVII) was cloned in vitro,with a point mutation from Lys to Ala in the position of 341 in the gene level.The cDNA fragments of mutational FVII (mFVII) and those of IgG1Fc were fused together with DNA ligase.After digestion,integration and sequencing,the fusion DNA was identified and transfected human embryonic kidney 293T cell packaging for re-mFVII/Fc lentiviral vector.After successful identification of vectors,detect the Ientiviral titer determination,bulk transfer after the determination of best MOI value of the third generation of hBMSCs,obseve the GFP expression with fluorescence microscope,have relative quantitative analyse of mRNA and protein expression of mFVII/Fc with RT-PCR and ELISA at different time points. Results In contrast with GenBank ID: AF 272774,the fusion gene matches exactly except the synonymous mutation,and the titer of packaging lentivirus was 2×108 TU/ml.Analyzed by Flow cytometry, indentification results of hBMSCs were as follows,CD+29(98.08%),CD+44 (97.63%),CD+34(0.31%) and CD+45(0.58%),respectively.The transfection efficiency of hBMSCs after 72 hours was (84±3)%,and the hBMSCs with mFVII/FC transfcetion have a large number of mRNA transcription and protein expression levels. Conclusions In this experiment we obtained a stable genetic vector with hBMSCs fusion gene expression successfully,which lay a foundation for the tissue factor study of prostate cancer targeting therapy and cancer gene therapy research.

OBJECTIVE:To investigate the clinical efficacy and safety of Cinobufacin capsules combined with radiotherapy in the treatment of median and advanced nasopharyngeal carcinoma. METHODS:Ninety-five patients with median and advanced naso-pharyngeal carcinoma in cancer center of our hospital during 2011-2012 were analyzed retrospectively,and then divided into con-trol group (57 cases) and observation group (38 cases) according to therapy plan. Control group received radiotherapy alone, while observation group was additionally given Cinobufacin capsule 0.5 g,tid,till the end of radiotherapy. A treatment course last-ed for 4 weeks. The prolonged radiotherapy were compared between 2 groups. Clinical response rate was observed in 2 group,and 1-year and 3-year survival rate and the occurrence of ADR were followed up. RESULTS:The proportion of prolonged radiotherapy was 89.47% in control group,which was significantly higher than 52.63% of observation group,with statistical significance(P<0.05). At the end of therapy and one month after therapy,clinical total response rates of observation group were 81.58% and 63.16%,which were significantly higher than 70.18% and 45.61% of control group,with statistical significance (P<0.05). The incidence of white blood cell lowering,hemoglobin lowering,local side effect of radiotherapy, nasopharyngeal dysphagia/odyno-phagia in observation group were significantly lower than in control group,with statistical significance (P<0.05). There was no statistical significance in survival rate between 2 groups in 1st-year and 3rd-year follow-up (P>0.05). CONCLUSIONS:Cinobu-facin capsule combined with radiotherapy can significantly relieve the side effects of radiotherapy in median and advanced nasopha-ryngeal carcinoma patients.

Objective To evaluate the reliability of lactate clearance rate in evaluating the efficacy of early fluid resuscitation in the patients with severe sepsis.Methods One hundred and forty-two patients with severe sepsis,aged 28-87 yr,were enrolled in the study.Isotonic crystalloid fluid was infused after admission to ICU to maintain central venous pressure ≥ 8 mmHg,mean arterial pressure ≥ 65 mmHg,central venous oxygen saturation (ScvO2) ≥ 70％,and urine output ≥ 0.5 ml·kg 1 ·h-1 within 6 h.The patients were divided into 2 groups according to the treatment outcome:survival group and death group.Immediately before fluid resuscitation and at 6,12 and 24 h after fluid resuscitation,blood samples were collected from the central vein for blood gas analysis and ScvO2 was recorded.Blood samples were collected from the peripheral vein for determination of the concentration of lactate and lactate clearance rate was calculated.Results Compared with survival group,the lactate clearance rate was significantly decreased at 6,12 and 24 h after fluid resuscitation,and no significant change in ScvO2 was found in death group.Conclusion Lactate clearance rate can evaluate the efficacy of early fluid resuscitation in the patients with severe sepsis.

Objective To analysis the clinical outcome at discharge and its risk factors of extremely preterm infants.Method To retrospectively analysis the clinical outcome at discharge and it's risk factors of extremely preterm infants (less than 28 weeks gestation) admitted from September 2008 to August 2014 in our Hospital.Result A total of 179 cases were enrolled.Survival rate was 59.2％ (106/179).Unfavorable outcome rate was 74.3％ (133/179),among them 73 cases died.The top five causes of death were severe bronchopulmonary dysplasia (BPD) (28 cases),Ⅲ ～ Ⅳ o intraventricular hemorrhage (IVH) (19 cases),sepsis (16 cases) and necrotizing enterocolitis (NEC) (6 cases).Among the 60 survivals with unfavorable outcomes,35 cases had either severe neurologic or ophthalmological sequela,and 25 cases had severe pulmonary sequela.Univariate analysis showed that,comparing with improved group,unfavorable outcome group had higher rates of not receiving prenatal steroids,placental abruption,male,small for gestation age,resuscitation with chest compression,admission age older than 72 hour,severe respiratory distress syndrome (RDS),without pulmonary surfactant (PS) usage,mechanical ventilation beyond 2 weeks and sepsis (P ＜ 0.05).Logistic regression analysis showed that those without prenatal steroids (OR =9.402,P =0.002),small for gestational age (OR =8.271,P =0.018),resuscitation with chest compression (OR =6.325,P =0.023),admission age older than 72 hour (OR =4.174,P =0.028) were independent risk factors for unfavorable outcome of extremely premature at discharge.Conclusion Extremely preterm infants have a higher rate of unfavorable outcome at discharge.Avoid small for gestational age,transfer properly and in time both in utero and after birth,and conduct prenatal steroids could improve their clinical outcome at discharge.

Objective To evaluate the efficacy of intracoronary,antologous bone marrow mononuclear cell transplantation on the patients with chronic heart failure.Methods Eighty patients with chronic heart failure were assigned into bone marrow cell transplantation group (n=48) and control group (n=32),with intmcomnary delivery of autologous bone marrow mononuclear cell suspension or not respectively,in addifion to conventional treatment.The left venlrieular ejection fractions (LVEF),the level of plasma brain natriuretic peptide (BNP),and the siges of myocardial perfusion defect were measured and compared with useing single photom emission computed tomography (SPECT) and echocardiography before and after the treatment.Results The one year follow-up results showed that LVEF increased from base-line (42.1±5.8)%,(43.9±6.7)% to (54.1±4.8)%,(49.8±7.7)% by echocardiography.The plasma levels of BNP decreased from base-line (699±101)ng/L,(687±95)ng/L to (305±78)ng/L,(399±89)ng/L,and the size of myocardial perfusion defect decreased from bese-line (26.8±8.6)%,(26.3±6.5)% to (15.8±4.3)%,(20.5±7.8)% in bone marrow cell transplantation group and control group respectively,the absolute changes were statistically significant.During and after the procedure of bone marrow cell transplantation group,there was no complication occurring. Conclusion Intmcoronary autologous bone marrow mononuclear cell transplantation for treating patients with chronic heart failure results in improvement in left ventricular systolic function and myocardial perfusion,and this therapy is safe and feasible.