Objective To investigate the influence of start time of continuous renal replacement therapy (CRRT) on the survivay and renal function recovery of patients with severe acute kidney injury (AKI)Methods Two hundred cases of severe AKI patients in the Traffic Hospital of Shandong Province from January 2011 to January 2014 were selected and divide into groups A of 1 phase,group B of 2 phase and group C of 3phase based on the CRRT starting time.The survivary and renal function recovery of 3 groups patients were compared.Results After treatment,the recovery rate of renal function,survival rate and mortality of patients in group A were 91.04％,92.5％ and 36.2％,significantly higher than that of group B(52.11％,73.6％ and 7.5％) and group C (29.03％,36.2％ and 63.8％),the differences were statistically significant (x2 =6.266,16.143,16.143;P＜0.05);and the recovery rate of renal function and survival rate of group A was higher than of group B and C (P＜0.05);and renal function index of groupA,B,C all significantly improved than before treatment,there were statistically significant differences (P＜ O.05).Conclusion Early CRRT treatment can significantly improve the survival rate of patients with SCKI,which would be good for patients to recover renal function.

Objective To measure the fractional anisotropy(FA) values in the language function areas(Broca area,Wernicke area and arcuate fasciculus)by diffusion tensor imaging(DTI),discussing the pathological characteristics in cerebral palsy children with language development delay.Methods DTI was performed in twenty-seven cerebral palsy children with language development delay(observation group) and 20 children with febrile seizures(control group) by Philips 3.0T MRI scanner.The FA values of Broca area,Wernicke area and arcuate fasciculus were measured at Philips Workstation.Results The FA values of control group in the left Broca area,Wernicke area and arcuate fasciculusis were higher than those of the right side,and the difference was statistically significant (all P ＜ 0.05).The FA value of observation group in the left Broca area was lower than that of the right side,and the difference was statistically significant (P ＜ 0.05);the FA values of observation group in the left Wernick area and arcuate fasciculusis were higher than those of the right side,but the difference was not statistically significance (all P ＞ 0.05).The FA values in the bilateral Broca area and arcuate fasciculusis of observation group were lower than those of the control group,the difference was statistically significant (P ＜ 0.05),those in the bilateral Wernick area were also lower than the control group,but the difference was not statistically significant (P ＞ 0.05).Conclusions The main pathogenesis for language development delay in cerebral palsy children was extensive damage in language functional areas,and Broca area and arcuate fasciculus were markedly impaired than Wernick area.

In order to obtain nucleotides aptamers bind to IgE, 80 bp nucleotides single-stranded DNA library containing 40 random nucleotides was designed and synthesized. Oligonucleotides that bind to human Cepsilon3-Cepsilon4 protein were isolated from ssDNA pools by the systematic evolution of ligands by exponential enrichment (SELEX) method using nitrocellulose filters as screening medium. Through the optimization of critical PCR and asymmetric PCR parameters including annealing temperature, cycles, and molar ratios of target protein and ssDNA etc, a suitable screening system was established. The aptamers of Cepsilon3-Cepsilon4 protein with high affinity and high specificity were identified by ELISA with biotin-streptavidin-horseradish peroxidase system, and its primary sequence and second structure were analyzed by DNAMAN package and DNA folding sever after being cloned and sequenced. Moreover, target protein was bound to one aptamer and another aptamer modified with biotion together forming a sandwich-like complex, which was captured in microwell to detect IgE concentration using the optimal combination in the sandwich method named enzyme-linked aptamers sorption assay (ELASA). The method could be used for the quantitative detection of human IgE, and whose sensitivity reached to 120 ng x mL(-1).

Objective To investigate the effect of co-expression of neuropilin-1(Nrp-1)and transforming growth factor-β(TGF-β)on regulatory T cell(Treg)-mediated immunosuppression. Methods CD4+CD25+Tregs were isolated from the spleens of male Balb/c mice. CD4+CD25+Tregs were blocked with various doses of Nrp-1 antibody(Ab-Nrp-1,0.5,5,10 μg/ml)for 24 hours with anti-CD3/CD28 and lipopolysaccharide(LPS)stimulation, and expression of Nrp-1 and TGF-βwas determined by flow cytometry. Meanwhile,CD4+CD25+Tregs were cultured with different doses of Ab-Nrp-1 for 1 hour,and co-cultured with CD4+CD25-T cell for 12,24 and 48 hours respectively,the proliferative activity of CD4+CD25-T cells was analyzed by microplate reader. Results Compared to control group,the expressions of Nrp-1 and TGF-β were significantly increased under anti-CD3/CD28 and LPS stimulation(both P<0.05),and treatment with Ab-Nrp-1 markedly inhibited the expression of TGF-β in a dose-dependent manner(P<0.05). The normal Treg had the potential to inhibit the proliferation of CD4+CD25-T cells(P<0.05),while various doses of Ab-Nrp-1 had the ability to reverse the immunosuppressive function of CD4+CD25+Treg in a dose-and time-dependent response manner,5μg/ml has the strongest ability,expecially after 24 hours. Conclusion Treg cell plays an important role in mediating immunosuppressive response via membrane associated TGF-β,and co-expression of Nrp-1 can markedly promote the immunosuppressive function.

Objective To investigate the FH/Wjd rat model of alcoholic liver disease.Methods Thirty-six 16-18 week old SPF grade FH/Wjd rats (male:female=1:1) were used in this study.The rats were divided into two groups randomly by body weight:water intake group and alcohol intake group.The rats took water or alcohol freely.16 weeks lat-er, ALT, AST, TBIL, TG, CHO in the serum and TG, GSH in the liver homogenate were detected.The expression of PPARαprotein in the liver tissue was detected by Western blot.The apoptosis rate of liver cells was assessed by flow cy-tometry.The pathological changes of liver tissue were examined using HE staining.Results Compared with the water in-take group, the serum TBIL and TG were significantly increased in rats of both sexes of the alcohol intake group, moreover, ALT and CHO of the female rats in the alcohol intake group were significantly decreased.TG in the liver homogenate in-creased obviously, while GSH in the liver homogenate showed a decreasing tendency.Hepatocyte apoptosis in rats of both sexes in the alcohol intake group showed an increasing tendency.The PPARαprotein expression was up-regulated obvious-ly, and the main pathological change in the liver tissue was microvesicular fatty degeneration.Conclusion Spontaneous long-term alcohol intake can induce liver injuries in FH/Wjd rats.

Aim To investigate the effect of Jinlida on cholesterol-related genes in skeletal muscle in fat-in-duced insulin resistance ApoE-/ - mice. Methods Ten male C57 BL/6 J mice were selected as normal group ( NF );50 male ApoE-/ - mice with a high-fat feeding after 16 weeks ( HF) were divided into model group, rosiglitazone ( LGLT ) , Jinlida low dose group ( JLDL, 0. 95 g · kg-1 · d-1 ) , Jinlida medium dose group ( JLDM, 1. 9 g·kg-1 ·d-1 ) , Jinlida high dose group (JLDH, 3. 8 g·kg-1·d-1), which were per-formed intragastric administration for 8 weeks. Oil red O staining of mouse skeletal muscle was used for fat ac-cumulation. Insulin receptor ( INSR) , insulin receptor body substrate-1 ( IRS-1 ) , low-density lipoprotein re-ceptor ( LDLR ) , cholesterol sensor ( SCAP ) mRNA and protein expression in mouse skeletal muscle were measured by quantitative reverse transcription PCR ( RT-PCR ) and Western blot. Results Compared with NF group, fasting blood glucose ( FBG) , choles-terol ( TC ) , triglyceride ( TG ) and low density lipo-protein cholesterol ( LDL-C ) of HF mice were signifi-cantly elevated, while high-density lipoprotein ( HDL-C ) significantly decreased ( P < 0. 05 ) . Compared with HF group, Jinlida group could reduce to varying degrees FBG, TC, TG and LDL-C in mice, and in-crease HDL-C ( P <0. 05 ) . Jinlida could downgrade fasting serum insulin ( FINS ) level, and improve the insulin sensitive index ( ISI ) ( P < 0. 05 ) . Jinlida could obviously improve skeletal muscle fat accumula-tion of mice. Compared with NF group, skeletal mus-cle INSR, IRS-1, LDLR mRNA and protein levels of HF group were significantly decreased ( P <0. 05 ) , while SCAP mRNA and protein level increased signifi-cantly (P<0. 05). Compared with HF group, Jinlida could increase to varying degrees INSR, IRS-1, LDLR mRNA and protein levels ( P < 0. 05 ) , and lower SCAP mRNA and protein levels ( P<0. 05 ) . Conclu-sion Jinlida can alleviate fat-induced insulin resist-ance in ApoE-/ - mice through regulation of cholester-ol-related gene expression.

OBJECTIVE To explore the effect of Lianhuaqingwen capsules ( LHQW ) on junction protein expression in mouse lung tissue of lipopolysaccharide (LPS)-induced acute lung injury ( ALl). METHODS 120 male mice were randomly divided into six groups: normal control, model, model+dexa-methasone 5 mg.kg-1 , model +LHQW 2, 4 and 8 g.kg-1 groups. Dexamethasone and LHQW were administered orally, once daily, for 7 d. 24 h after the last administration, LPS solution was instilled into the tracheas of mice except the normal control group to prepare the mouse model of ALl. 24 h after the establishment of the ALl model, the mice were sacrificed and the pathological changes in the mouse lung tissue were observed by optical microscopy and ultrastructure of alveolar epithelium was observed by transmission electron microscopy. The cell percentage of positive expression of tumor necrosis factor-α(TNF-α) in the peripheral blood T lymphocytes was detected by flow cytometry. The expressions of con-nexin 43 ( Cx43), occludin and zonula occludens protein-1 ( ZO-1) in lung tissues were detected by immunohistochemistry. RESULTS Under the light microscope, the mouse lung of model group showed a large amount of inflammatory cell infiltration and alveolar wall thickening. Compared with model group, inflammatory cell infiltration was reduced in model+dexamethasone, model+LHQW 2,4 and 8 mg.kg-1 groups. Under the electron microscope, the mouse alveolar epithelial cells of model group showed injury. Compared with model group, the damage was reduced in model+dexamethasone, and model+LHQW 2, 4 and 8 mg.kg-1 groups. The cell percentage of TNF-α positive expression in peripheral blood T lympho-cytes in normal control, model, model+dexamethasone, model+LHQW 2,4 and 8 mg.kg-1 groups was (3.6±0.9)%, (6.4±0.8)%, (2.8±0.7)%, (4.7±1.6)%, (4.0±1.5)% and (3.6±1.2)%, respectively. The percentage in model group was obviously higher than that in normal control group( P<0.01), but was lower in the four drug treatment groups than in model group(P<0.05, P<0.01). The expression of Cx43, occludin and ZO-1 in lung tissue of model group was lower than that of normal control group(P<0.01), but higher in model+dexamethasone, model + LHQW 4 and 8 mg.kg-1 groups than in model group(P<0.05). CONCLUSION LHQW may alleviate ALl induced by LPS and play a protective role by inhibiting inflammatory cell infiltration and improving protein connection expression in alveolar epithelial cells and pulmonary vascular endothelial cells.

Objective To compare the differences of lung pathological changes of acute lung injury in mice in-duced by lipopolysaccharide ( LPS) and graphite particles, and to explore the possible mechanisms of acute lung injury in-duced by fine particles of different origins.Methods 140 male specific-pathogen-free Kunming mice weighing 18-20 g were randomly divided into 7 experimental groups, in addition to the normal control group.The experimental groups were treated by intratracheal instillation of LPS solution or graphite powder suspension in different doses, respectively, to induce acute lung injury in the mice.The mortality of the mice was observed, and pathological changes of the lung tissues were ex-amined by light and transmission electron microscopy.Western blot was used to detect the protein expression of neutrophil elastase ( NE) in lung tissues , and real-time quantitative PCR was used to detect mRNA expression of monocyte chemotac-tic protein-1 ( MCP-1) in the lung tissue .Results Compared with the normal control group, some pathological changes were observed in the lung tissues of the groups L ( LPS) and G ( graphite) .There were numerous macrophages in the lung tissues in the group G mice, and exudate, mainly neutrophils, in the lung tissues of the group L.The NE protein expres-sion in the lung tissue was significantly higher than that of the normal control group ( P<0.05) , and there was also a sig-nificant difference between the groups L and G (P<0.05).The MCP-1 mRNA expression in lung tissues was higher in the control group (P<0.01), and there was also a significant difference between the groups L and G (P<0.01).Conclu-sions Diverse types of particulate matters induce different pathological changes in the lungs, therefore the mechanism may also be different in the inflammatory responses.It means that the lung injuries caused by fine particles of mixed composition may have complex mechanisms.

Aim To construct the stable hFcεRIα/RBL-2H3 cell line expressing human FcεRIα( hFcεRIα) . Methods The human FcεRIα gene was obtained by RT-PCR and cloned into the eukaryotic expression vector pCI-neo. Then, the pCI-neo-hFcεRIα vector was transfected into RBL-2H3 cells by lipo-somes, and the transfected cells were screened through G418 fil-tration subsequently. Finally, RT-PCR, Western blot and immu-nofluorescence assay were used to determine the result of trans-fection. Results According to the optimized transfection param-eters, the transfection efficiency reached 75. 38%. The results of Western blot, immunofluorescence and RT-PCR showed that hFcεRIα could be expressed in RBL-2H3 cells successfully. Conclusion HFcεRIα/RBL-2H3 cells were successfully con-structed,which will be the experimental basis for further study on the mechanism of IgE/FcεRI and drugs for allergy diseases.

Objective To observe the effect of emotional resilience group training on fatigue and sleep quality of patients with gastrointestinal cancer.Methods A total of 321 hospitalized patients with gastrointestinal cancer were randomly divided into experimental group (160 cases) and control group (161 cases) by random number table.Two groups of patients were treated with routine nursing care.In addition,the experimental group was given 8 weeks of emotional resilience group training once a week.The effect of intervention was assessed by the cancer fatigue scale (CFS) and Pittsburgh sleep index (PSQI) before and after the intervention.Result There was no significant difference in CFS and PSQI between the two groups before intervention (t=0.18,1.82,P＞0.05).After intervention,there was no significant difference in the total scores and each dimension scores of CFS and PSQI in the control group (P＞0.05).The total scores of CFS and PSQI in the intervention group (13.72± 1.33 and 10.62± 1.01) were significantly lower than those before intervention (25.35 ± 2.07 and 17.38 ± 2.69).The dimensions of CFS,sleep quality,sleeping time,sleep disorder,hypnotic drug use and daytime dysfunction were significantly lower than those before intervention (P＜0.01,P＜0.05).After intervention,there were significant differences in scores of CFS,somatic,cognitive and emotional dimensions between the two groups (t=18.21,-36.94,-13.17,-6.17,P＜0.01),and the scores of PSQI,sleep quality,sleeping time,sleep disorder,hypnotic use and daytime dysfunction were statistically different between the two groups (t=19.96,-82.86,-16.59,-9.39,-28.00,-9.25,P＜ 0.01).Conclusion Emotional resilience group training can effectively reduce the fatigue of patients with gastrointestinal cancer and improve sleep quality.