Humans ; Hyperamylasemia ; physiopathology ; Infant, Newborn ; Neonatology

Objective:To observe the effect of Allicin in cardial fibroblasts (CFs) proliferation and Collagen secretion,and to explore its role on TLR4/NF-κB signal pathway.Methods:CFs of neonatal Wistar rats were isolated and cultured ,then was stimulated with AngⅡ.CFs proliferation was measured by thiazolyl blue ( MTT) assay.The expression of collagenⅠ,collagenⅢ was measured by ELISA.mRNA expression of TLR4 and NF-κB were detected by reverse transcription-polymerase chain reaction ,protein expression of TLR4 and NF-κB were detected with Western blot.Results: Allicin could reduced MTT value of cardial fibroblasts ( P<0.01 ) , and inhibited expression of collagenⅠ,collagenⅢ(P<0.01),which in a dose-dependent manner.Allicin could reduced mRNA expression of TLR4 and NF-κB and protein expression of TLR 4 and NF-κB in CF induced by Ang Ⅱ ( all P<0.01 ) .Conclusion: Allicin can inhibit Myocardial fibrosis ,which mechanism is possible by inhibiting TLR 4/NF-κB signal pathway.

Objective To evaluate the efficacy of naloxone used in the postoperation of cerebral tumor.Methods Eighty patients were randomly assigned to receive (treated group:40 patients) or not re- ceive (control group:40 patients) naloxone.Both the two groups accepted the conventional therapy.Re- sults After operation,the content of?-EP,ET decreased continuously but the one of the treated groups was more obviously than that of the control groups (P

The complete genomes of more cyanobacterial strains have been completed for sequence, and genetic engineering of cyanobacteria has evolved in the post-genome era. Since Kaneko and colleagues had completed the sequence for the complete genome of Anabaena sp. PCC 7120 in 2001, functions of some genes in this genome, including fructose-1,6-bisphosphate aldolase (FBA) gene, have been predicated using the method of bioinformatics. However, little information is available regarding whether this gene can encode FBA and its product characteristics of related enzyme. Here, to explore this information, the predicted II-FBA gene-encoding region in Cyanobase database was cloned by PCR method and then ligated into pET-32a to generate the expression vector, pET-FBA-II. The results of SDS-PAGE indicated that the expression level of the expected target polypeptide was approximate 23.4 percent as compared to total protein and the molecular weight is about 40 kDa as compared to the protein molecular marker. The results of enzyme activity analysis showed that the activity of II-FBA was ~11.8 U per mg protein and owned a standard activity of II-FBA. To sum up, the results not only prove the functional prediction of this II-FBA gene from the Cyanobase database, but also provide the important conditions for further studying its physiological and biochemical characteristics and functions of the gene expression product.

The changes of bioactive constituents were analyzed for Puhuang-Wulingzhi before and after compatibility and the antiplatelet and antithrombin activitives were evaluated in order to elucidate the scientific and reasonable of Puhuang-Wulingzhi compatibility. UPLC-QTOF-MA-Markerlynx, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis were used for data analysis and tracking changes of chemical composition during the decocting process. In vitro platelet aggregation induced by ADP, thrombin time(TT) and prothrombin time (PT) were investigated for Puhuang-Wulingzhi before and after compatibility. The results showed that significant differences were found between the mixed decoction and codecoction of Wulingzhi and Puhuang. Five compounds changed obviously were identified as typhaneoside, naringenin, isorhamnetin-3-O-ruinoside, quercetin-3-O-neohesperidoside, kaempferol-3-O-neohesperidoside. The codecoction, comparing with the single decoction, was more significant in antiplatelet aggregation and could prolong thrombin time. In the same crude drug dose, the thrombin time (TT) elongation were greater. These data could provide references for elucidation of bioactive components for this herb pair.
Animals ; Antithrombins ; chemistry ; pharmacology ; Blood Platelets ; drug effects ; physiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Humans ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rabbits ; Thrombin Time

Objective Establish rabbit VX2 soft tissue tumor model,and treat it with percutaneous ethanol injection(PEI)under the guidance of magnetic resonance imaging.Make ready for the therapeutic evaluation with functional magnetic resonance imaging. Methods Fifteen healthy New Zealand white rabbits were included in this study.0.2 mL tumor tissue suspensions were injected into the rabbits’posterior limb.14 days later,all rabbits were underwent conventional MRI examination.PET were performed to all the tumors under the guidance of MRI in the next day of the examination.T2 WI was used as guidance and monitoring means.MR com-patible puncture needle with lateral hole was stabed into the lesion center,and inj ected anhydrous ethanol according to the volume of tumors’diameter (1 mL/cm )slowly.the tumors signal characteristics,morphological feature and pathological feature were ob-served pre and post-operation.Results All of the 1 5 rabbits were established soft tissue tumor model successfully;the success rate is 100%.The tumors were oval or round,3-4 cm in diam.MRI scanning showed low signal on T1 WI and high signal on T2 WI be-fore PEI.PEI was performed to all the tumors under the guidance of MR successfully with 3.5 mL ethanol injected into the tumors in average.T2 WI could monitor the ethanol in dispersion and distribution within the tumors clearly.Histologically,tumors were composed of large,uniform,oval/round cells arranged in solid nests which was intensive in the periphery of tumors.Necrosis tissue was apparent in the center of the tumors.10 days after operation,most tissue in the periphery of tumors was coagulative necrosis , only a few tumor cells left.Ranges of necrosis in the tumors center were obviously increased compared with pre-operation.Conclusion Rabbit VX2 tumor of soft tissue model is suitable for the therapeutic evaluation of tumor .It is an animal model which has the characteristic of simple to operate and high rate of suc-cessful.MR T2 WI can monitor the ethanol in dispersion and distribution within the tumors clearly.It is a good guidance and monitoring imaging method of tumor ablation.

Objective To establish an inactivated viral particle-based ELISA for the detection of antibodies against Middle East respiratory syndrome coronavirus (MERS-CoV) in serum samples collected from a MERS-CoV associated case. Methods Serum samples were collected from 10 newborns and 40 healthy adults. A viral particle-based ELISA was established by using the inactivated MERS-CoV virions as antigen. The levels of IgM and IgG antibodies in the serum samples were detected by the established ELISA and the cut-off values for positive detection were determined. Then the inactivated MERS-CoV virion-based ELISA was used to detect the antibodies against MERS-CoV in 5 serum samples collected from the first im-ported MERS case in China. Results The cut-off values of IgM and IgG antibodies in serum samples for ELISA were determined to be A450 readings of 0. 32 and 0. 42, respectively. The titers of IgM and IgG anti-bodies in serum samples collected at early admission to hospital from the first imported MERS case in China were both 1 ︰ 40. Seroconversion occurred 2 weeks after his admission to hospital with the titers of IgM and IgG reaching to 1 ︰ 320. Conclusion The inactivated MERS-CoV virion-based ELISA was established successfully and could be used for the detection of serum antibodies (IgG and IgM) in MERS associated cases.

Objective To investigate the effect of non invasive cardiac output monitoring(NICO)system in pig model with acute respiratory distress syndrome(ARDS),and to provide experimental basis for clinical application. Methods Eleven anaesthetized and ventilated ARDS male pig models were induced by intravenously infusing 0.2 mL/kg oleic acid. Lung recruitment was condocted by pressure control ventilation on pigs with ARDS. The optimal positive end-expiratory pressure(PEEP)was determined by optimal dead space fraction〔the ratio of dead space to tidal volume(VD/VT)〕. Cardiac output(CO)was determined by NICO,the respiratory function was monitored, and the VD/VT,dynamic compliance(Cdyn),oxygenation index(PaO2/FiO2),the volume of alveolar ventilation(Valv) and arterial blood oxygen saturation(SaO2)were recorded before infusing oleic acid,after stabilization of ARDS model and at optimal PEEP level,and the intrapulmonary shunt fraction(Qs/Qt)was calculated. CO was also determined by application of pulse indicated continuous cardiac output(PiCCO),and the linear regression analysis between CO determined by NICO and CO determined by PiCCO was conducted. Results Seven experimental ARDS pigs model were successfully established. The optimal PEEP identified by the lowest VD/VT method was(15.71±1.80)cmH2O (1 cmH2O=0.098 kPa). Compared with before infusing oleic acid,VD/VT and Qs/Qt after stabilization of ARDS model were significantly increased〔VD/VT:(72.29±8.58)% vs.(56.00±11.06)%,Qs/Qt:(21.04±15.05)%vs.(2.00±1.32)%,both P<0.05〕,and SaO2 and Valv were significantly decreased〔SaO2:0.888±0.108 vs. 0.999±0.053,Valv(mL):92.06±35.22 vs. 146.11±45.43,both P<0.05〕. VD/VT,Qs/Qt,SaO2 and Cdyn at optimal PEEP level were improved to the levels before infusing oleic acid〔(61.07±9.30)%,(3.21±6.10)%, 0.989±0.025,(117.14±41.14)mL〕. Cdyn and PaO2/FiO2 after stabilization of ARDS model were significantly lowered compared with those before infusing oleic acid〔Cdyn (mL/cmH2O):14.43±5.50 vs. 38.14±6.72, PaO2/FiO2 (mmHg, 1 mmHg=0.133 kPa):78.71±23.22 vs. 564.37±158.85, both P<0.05〕. Cdyn and PaO2/FiO2 at optimal PEEP level〔(19.71±4.86)%,(375.49±141.30)mmHg〕were elevated compared with the levels after stabilization of ARDS model(both P<0.05),but still lower than those before infusing oleic acid(both P<0.05). Compared with the levels after stabilization of ARDS model,CO at optimal PEEP level showed obvious decrease from(4.18±2.46)L/min to(3.95±2.69)L/min without significant difference(P>0.05). There was linear correlation between CO determined by NICO and CO determined by PiCCO(r2=0.925,P<0.001). Conclusions NICO technique provides a useful and accurate non invasive estimation of CO and respiratory function.VD/VT provided by NICO can titrate the optimal PEEP in patients with ARDS.

Aged ; B-Lymphocytes ; pathology ; Histiocytic Sarcoma ; complications ; Humans ; Lymphoma, B-Cell ; etiology ; Lymphoma, Large B-Cell, Diffuse ; etiology ; pathology ; Male

This study was aimed to investigate the protocol in vitro to incubate the dendritic cell (DC) derived from peripheral blood monocytes using serum-free medium X-VIVO 20. Peripheral blood monocytes from healthy donors were treated with 100 ng/ml GM-CSF and 500 U/ml IL-4, respectively. After cultivation for 6 days, they were treated with 100 ng/ml calcium ionophore A23187. After cultivation for 24 hours the cellular morphology was observed under invert microscope, the surface markers were analyzed by flow cytometry, the proliferation of allogenetic T cells was detected by MTT colorimetry, the specific cytotoxicity of T cells primed with DC was examined by MTT assay. The results showed that in all three groups with serum-free, fetal calf serum (FCS) and human AB serum mediums, cells displayed characteristic morphological features of DC. Simultaneously CD14 expression was decreased, and CD83, HLA-DR and CDw123 expression were increased on these cells. In addition, DCs cultured with these methods could evidently stimulate the proliferation of allogenetic T cell. As compared with the two controls of serum containing groups, the cultured cells in the serum-free groups showed almost the same allo-stimulatory capability and cellular morphology and surface markers, and T lymphocytes primed with the culture-derived DC exhibited the similar killing activity to K562 (P > 0.05). It is concluded that there is no significance in DC numbers, morphology, epitope and ability to stimulate the proliferation of allogenetic T cells between DC induced by serum-free X-VIVO 20 medium and DC induced by serum-contained medium. DC cultured and induced by serum-free medium is worth using in practice widely.
Cell Culture Techniques ; Culture Media, Serum-Free ; Dendritic Cells ; cytology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interleukin-4 ; pharmacology ; Monocytes ; cytology ; Recombinant Proteins ; T-Lymphocytes ; cytology ; T-Lymphocytes, Cytotoxic