Objective To investigate changes in NF ?B signal pathway in PAM stimulated by lipopolysaccharide(LPS) in vitro , and to explore the molecular pathological mechanism of acute lung injury(ALI). Methods After PAM were stimulated by LPS, the changes in expression of IKK mRNA, activation of NF ?B, degradation of I?B, and secretion of TNF ? in PAM were measured at 0, 15min, 30min, 1h, 2h, and 4h by in situ hybridization, electrophoretic mobility shift assay (EMSA), and enzyme linked immune absorbing analysis (ELISA), respectively. Results The expression of IKK ? mRNA was increased 15min after LPS stimulation and reached the peak at 30 min, then returned to the base line after 1 hour. The changes in I?B ? mRNA were opposite. The activity of NF ?B was increased 15min after LPS stimulation, peaking at 1 hours, and returned to the pre stimulation level after 2 hours. The content of TNF ? was increased initially at 30min, reached the peak at 1 hour, and gradually returned to the pre stimulation level in 2～4 hour. Conclusion The transduction pathway of activation of IKK ? degradation of I?B/activation of NF ?B/synthesization of TNF ? might play a critical role in the molecular pathological mechanism of LPS induced ALI.

Objective Beijing plans to run a DRGs-PPS pilot.Cost estimation of each DRG group needs a set of scientific reasonable standards for inpatient charge category.MethodsResource consumption accounting Classify charge item by practitioners,services,resources; medical consumables are singled out to emphasize the value of medical personnel's services; some of old categories are subdivided in order to be compatible with new categories.Results 19 old charge categories of patient discharge chart turn to 31 new items by dividing medical service into treatment,surgery,nursing,imaging,examination,management,paramedic,pharmacy,etc.ConclusionRegulating the charge categories of services,medical data is more consistent by using the same statistical coverage.Also it accurately present the incoming of all operationstreatment,surgery,nursing,imaging,examination,management,paramedic,pharmacy,etc.It gives government directions of decision-making to adjust the weight of DRG groups.

Objective To explore the neural basis of the visual illusory motion using the event-related potential(ERP).Methods One hundred trials(each duration 2 000 ms)of a visual illusory motion figure,rippling wheat pattern made by Akiyoshi Kitaoka,and a visual static figure made of modified rippling wheat pattern were randomly presented with equal probability.Ten healthy right handed undergraduate students as subjects were asked to judge the stimulus whether motional or not.The EEG was recorded from 128 scalp sites using with electrodes mounted in HydroCel GSN cap(Electrical Geodesics Incorporated,Oregon,USA).Results 1)The C1 component of ERP could be evoked by both visual illusory motion pattern and static pattern in POz.The peak of C1 component was presented about 75 ms after each stimulus.C1 component evoked with visual illusory motion was negative,but it's static pattern to be positive.2)P100 and P200 components could be obviously evoked by both kinds of stimuli,but in O2 the amplitudes of P100 evoked by different stimuli were significant different and in T3 the amplitudes of P200 were significant different too.Conclusion The visual illusory motion is formed at the primary visual cortex.It may be related to the organized mode of perception.

Objective: To investigate the targeting and anti-tumor ability of the tumor vessel-specific antibody ScFvH1 selected from phage-ScFv library, and to discuss the application of the antibody in clinical diagnosis and therapy of cancer. Methods: The ScFvH1 gene was inserted into pET-28a(+)/EGFP vector containing green fluorescent protein(GFP) gene and pTIG-Trx vector containing thioredoxin gene; the products were then expressed in E.coli and purified by using Ni-NTA. Tumor-bearing mice model was established by subcutanuous injection of cervical cancer cell line HeLa. The mice were injected with purified ScFv-EGFP fusion protein through vena caudalis and the GFP signals were observed by fluorescent microscope to evaluate the targeting ability of the antibody. Meanwhile, the mice model also received intratumoral injection of purified ScFv-EGFP fusion protein to evaluate the anti-tumor effect of the antibody. Results: Soluble ScFvH1 gene and ScFvH1-EGFP protein were successfully expressed in E.coli; a single band was showed in SDS-PAGE after the purification by Ni-NTA. We found that ScFvH1-EGFP fusion protein was enriched to tumor tissues, but there was only weak fluorescent signal when EGFP protein was injected. No EGFP signal was observed in the lung of tumor-bearing mice. Tumor inhibition experiment showed that the tumor growth in the antibody treatment group was similar to that of the PBS control group. Conclusion: The tumor vessel-specific antibody ScFvH1 selected from phage-ScFv library can specifically target tumor vessels, but it has no obvious inhibitory effect on tumor growth. Our findings pave a way for antibody in cancer diagnosis and treatment.

Objective To investigate the role of endogenous histamine in Parkinson disease. Methods 6-OHDA-lesioned rats were prepared by the conventional mothod,and in the meantime a group of rats were administrated with ?-fluoromethylhistidine(?-FMH),an irreversible inhibitor of histidine decarboxylase(HDC),via intracerebroventricular injection(12.5 ?g or 25 ?g,i.c.v.) for seven days.On the 7 day,the apomorphine-induced turning behavior and were detected, and the immunoreactivity of dopaminergic neurons in the substantia nigra pars compact(SNc) and histaminergic neurons in the tuberomammillary nucleus(TMN) were evaluated by tyrosine hydroxylase(TH) and HDC immunohistochemistry,respectively.Additionally,the level of dopamine in striatum was determined with high performance liquid chromatography(HPLC). Results ?-FMH(25 ?g,i.c.v.) significantly reduced the turning behavior and prevented the loss of dopaminergic neurons in the SNc,and slightly increased dopamine level in the striatum.Whereas,the immunoreactivity of histaminergic neurons in the TMN of hypothalamus in both the 6-OHDA lesioned and the ?-FMH treated rats was not changed.Conclusion Endogeneous histamine may involve in the pathological processes of PD.However,the histaminergic neurons are not involved in PD.

Objective To investigate the effect of glutamine on energy intake and prognosis of severe burn patients in differ-ent pathway.Methods Using retrospective study method,we brought the patients into our study from October 2010 to April 2014 in accordance with the inclusion criteria.Patients who were given the Gln before 5 days after injury through gastric bowel were brought into the EN group,others who were given the Gln after 5 days after the injury through vein were brought into PN groups. A total of 66 patients were included in this study,with 31 cases of EN group and 35 cases of PN group.Total energy intake,external and internal energy intake,nutrients heating,energy intake/energy consumption ratio,blood glucose control,insulin use and viscera damage(the blood urea nitrogen,creatinine,alanine aminotransferase,aspartate aminotransferase,total bilirubin,creatine kinase,lac-tate dehydrogenase),ICU days,hospital stays and mortality were observed in 1,2,3,4 weeks after injury.Results Within a month after the injury,the energy intake of patients in EN group were more than PN group,especially the energy from intestinal canal.Be-sides,the ICU days of patients in EN group were shorter than patients in PN group (all P <0.05).Other indicators of two groups of patients were no significant statistical difference(P >0.05).Conclusion Supplement of Gln may be more conducive to improve the intestinal function in patients with burns may be more conducive to improve the intestinal function in patients with burns,im-prove the degree of tolerance of enteral nutrition,increase energy intake and conducive to improving the prognosis of patients.

Objective:To obtain phage-displayed ScFv library targeting tumor tissues and to screen for antibodies specifically binding to tumor vessels using in vivo phage display,so as to lay a foundation for diagnosis and treatment of cancer.Methods:The membrane proteins were extracted from the specimens of esophageal carcinoma,stomach carcinoma,brain cancer,lung cancer,and spinal cord tumor.The recombinant phage-antibody system was used to construct a single-chain Fv fragment(ScFv)cDNA library from the total RNA of the BALB/c mice immunized with purified membrane protein.The specific primers of VH and VL were used to amplify the cDNA of VH and VL,respectively,which were then assembled into ScFv gene with a specially constructed linker DNA.The ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated samples were transformed into competent E.coli TG1.The transformed cells were infected with M13KO7 helper phage to yield recombinant phage.Using the animal model of human cervical carcinoma(HeLa cells),sepecific phage-ScFvs were selected by phage displaying and panning in vivo.After four rounds,24 phage-ScFvs,which were identified by PCR,were analyzed immunohistochemically.The ScFvs expressed in the tumor tissue slices and negative in control kidney tissue slices were sequenced.Results:Tomors-bearing animal models were established with 7 different kinds of carcinoma cell lines in BALB/c nude mice.It was found that inoculation with HeLa cells resulted in most satisfactory tumorigenesis in nude mice.A ScFv library of 1.6?106 was obtained and a tumor vessel specific phage-ScFv named ScFvH1(VH-linker-VL)was selected from the library.Conclusion:A tumor targeting ScFv library has been successfully constructed and a tumor vessel-specifrc antibody has been identified from the library,which provides a new way for the early diagnosis and therapy of cancer.

Objective To analyze the alterations of serum protein in ESCC,compare alterations of serum protein with and without LM. Methods Serum samples were collected from 64 ESCC patients before operation and 60 cases with gender and age-matched healthy controls,special serum protein or peptide spectra was determined by SELDI-TOF-MS measurement after treating the sample onto weak cation exchange (WCX2) protein chip for each case. The serum protein profiles were compared by Biomarker Wizard Software between the ESCC patients and healthy controls, and among ESCC patients stratified according to gender, age, location of tumor, size of tumor, infiltration and with or without LM. Results (1)120 protein peaks were detected at the molecular range of 0 to 50000 in comparing of ESCC patients and healthy controls. 31 significantly different peaks were found between ESCC patients and healthy controls (P <0.05), 10 peaks were selected(P<0.01). (2) One significantly different protein peak (m/z 4174) was detected between T1 and T3, T4 (P<0.05). (3) There were three significantly different protein peaks (m/z 3970,4174 and 4277) between with LM and without LM (P<0.05).The peak (m/z 4174) was shared by two groups above. (4) No significant different protein was found when patients stratified according to gender, age, location of tumor and size of tumor. Conclusion Significant difference exists in serum proteins between ESCC patients and healthy controls. There are statistical difference exists in serum proteins between T1 and T3, T4, with LM and without LM. This difference is less than between ESCC patients and healthy controls. Some commonness is existed in serum protein fingerprint for patients with serious infiltration and with LM.

ObjectiveTo observe the effect of Astragalus membranaous on angiotensinⅡ (AngⅡ)-induced transform-ing growth factor β1 (TGF-β1) production of cardiac ifbroblasts.Methods Cardiac ifbroblasts were culturedin vitro. Cells were allocated into 3 groups: control group, Astragalus membranaous groups (50, 100, 200 mg/ml), Ang II group (10-7 mol/L) and AngⅡ/Astragalus membranaous groups (50, 100, 200 mg/ml). The proliferation of each group was tested by methyl thiazolyl tetrazolium method. TGF-β1 was measured by ELISA.Results The proliferation of cardiac ifbroblasts had signiifcant difference between each groups (F=71.84,P=0.000). The proliferation of cardiac ifbroblasts with Ang II stimulation was higher than that of cells without Ang II stimulation (P<0.05). Astragalus membranaous inhibited Ang II-induced cardiac ifbroblasts proliferation dose dependently (P<0.05). The TGF-β1 production had signiifcant difference between each groups (F=786.81,P=0.000). The TGF-β1 production in AngII/astragalus membranaous groups was lower than that in Ang II group (P<0.05). The TGF-β1 production in Ang II group was the highest, and had signiifcant difference as compared to other groups (P<0.05). Astragalus membranaous inhibited Ang II-induced TGF-β1 production dose dependently (P<0.05).Conclusions Ang II can stimulate the proliferation of cardiac ifbroblasts, and promote the TGF-β1 production. Astragalus membranaous can inhibit the proliferation of Ang II-induced cardiac ifbroblasts, and reduce the TGF-β1 production of cardiac ifbroblasts.

Objective To investigate the expressions of Wnt2 and β-catenin in Doxorubicin (DOX)-induced myocardial injury and to explore their roles in myocardial cell apoptosis.Methods Cardiomyoblast cells were damaged by different concentrations of DOX(1 mg/L,2 mg/L,3 mg/L,4 mg/L) for 72 h.The effect of different concentrations of DOX on cardiomyocyte growth curve was detected according to the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-h-tetrazolium bromide(MTT) assay.DOX(1 mg/L) was used to induce the model of cardiomyoblast cell injury.Cardiomyocytes were divided into 4 groups:group A:DOX-injured cardiomyocytes for 12 h ;group B:DOX-injured cardiomyocytes for 24 h ; group C:DOX-injured cardiomyocytes for 48 h; group D:normal cardiomyocytes.The expressions of Wnt2,β-catenin and p53 were observed by Western blot and reverse transcription polymerase chain reaction(RT-PCR) at the time point of 12 h,24 h and 48 h.Results DOX significantly inhibited cardiomyocyte proliferation in a dose dependent fashion.The protein and mRNA expressions of Wnt2 increased in the DOX-induced myocardial injury group compared with the group D,with statistical significance (F =224.115,P ＜ 0.05) ;The expressions of β-catenin,p53 were significantly increased compared with the group D,and the higher expression appeared with the time extending(F =188.145,231.927,all P ＜ 0.05).Significantly positive correlation between Wnt2 and β-catenin expression was observed(r =0.940,P ＜ 0.05).Conclusions These findings suggest that Wnt2/β-catenin signaling pathway may play important roles in the cardiovascular disease and be useful for exploring the molecular mechanism of myocardial injury..