AIM: To study the technological parameters of the purification process of total flavonoids of Crataegus leaves with HPD-600 macroreticular resin. METHODS: Using orthogonal experiments,the adsorptive characteristics and elutive parameters of the process were studied by taking the elutive effects and purification ratio of total flavonoids of Crataegus leaves. RESULTS: Two significant factors which were the concentration of crude flavonoids solvent and elutant component were observed. CONCLUSION: The microwave-assisted extraction and purification process with macroreticular resin is employed,the content of total flavonoids is 80.75%, the rutin content is (1.38)%, The macroreticular resin can be used in the purification of flavonoids of Crataegus leaves.

Objective To explore the effect of triperygium wilfordii polyglucoside(TWP) on the podocytes of rats with diabetes nephropathy(DN). Methods One hundred SD rats were randomly divided into normal control group (Group A),DN group (Group B),TWP group(Group C).TWP group was divided into 3 subgroups(Ca,Cb,Cc)according to the different doses 4,8,16mg·kg-1·d-1,respectively.Rats in DN group and TWp group were given streptozocin(STZ)by intraperitoneal injection to establish animal model of diabetes.After 12 weeks,24 h urinary protein excretion rate(UAER),BUN,Scr,white blood cell(WBC),blood glucose(Glu),and kidney weight (KW)/body weight(BW)were determined.The renal pathological changes were evaluated by HE staining.The structural change of podocytes was observed by transmission electron microscope.The expressions of nephrin and podocin were evaluated by immunofluorescence staining. Results (1)Compared to group A,Scr,BUN,Glu,KW/BW,and UAER were significantly higher(P＜0.05)in group B and group C.Whereas the elevated liver enzymes and the decreased WBC were presented in group Cc (3/20). Compared to group A, the protein expressions of nephrin and podocin in nephridial tissue were lower, and the significant differences of pathomorphology in glomerulus,tubules and podocytes were observed in group B and group C. (2) Compared to group B, KW/BW and UAER were lower in group C (P＜0.01); the protein expressions of nephrin and podocin were higher in nephridial tissue; the pathomorphological improvements were exhibited in glomerulus,tubules and podocytes in group C, paralleled with the increase of TWP dose (P＜0.05 or P＜0.01).Conclusions TWP may exert the protective effect on podocytes in diabetic nephropathy rats,dependent on the dose of TWP. The mechanism may be associated with the up-regulatied expressions of nephrin and podocin.

Background Research demonstrated that alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid GluR2 (AMPA-GluR2) is associated with amblyopia.It has been shown that levodopa and cytidine diphosphate choline can improve visual function of amblyopic children,but the mechanism is unclear.Objective This study was to explore the possible effects of levodopa and cytidine diphosphate choline on amblyopia.Methods Monocular deprivation (MD) animal models were created in 60 2-week-old SD rats by monolateral eyelid suturing and observed for 31 days and reared in natural light together with 15 other matched normal healthy SD rats.The models were randomly divided into the MD group,levodopa group,cytidine diphosphate choline group and normal saline control group,with 15 rats for each group.40 mg/kg of levodopa,80 mg/kg of cytidine diphosphate choline,I ml normal saline were given to the rats,respectively,for 28 consecutive days.Expressions of the AMPA-CluR2 protein and AMPA-CluR2 mRNA in the rat visual cortex were detected by immunohistochemistry,Western blot and real-time fluorescence quantitative PCR.Use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The expression values of the AMPA-GluR2 protein (AMPA-GluR2/β-actin) and AMPA-GluR2 mRNA (2-△△Ct) were significantly lower in the MD group than those of the normal control group (protein:0.32 ± 0.02 vs.0.64 ± 0.05,t =13.287,P＜0.05 ;mRNA:0.30±0.01 vs.0.84±0.03,t=38.184,P＜0.05).Those in the levodopa group were significantly increased in comparison with the normal saline solution group (protein:0.59 ±0.04 vs.0.33 ±0.03,t =11.628,P＜0.05 ; mRNA:0.71±0.06 vs.0.33 ±0.02,t =13.435,P＜0.05).The expression values of the AMPA-GluR2 protein and AMPA-GluR2 mRNA were significantly increased in the cytidine diphosphate choline group compared with the normal saline solution group (protein:0.52 ± 0.04 vs.0.33 ± 0.03,t =8.497,P ＜ 0.05 ; mRNA:0.48± 0.04 vs.0.33 ± 0.02,t =7.500,P＜0.05).Conclusions AMPA-GluR2 is associated with the plasticity of visual development.Levodopa and cytidine diphosphate choline may improve visual function by down-regulating the expression of AMPA-GluR2 in the visual cortex.

Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/diagnosis* ; Death, Sudden/etiology*

Objective To investigate effects of xeroderma pigmentosum B(XPB) gene on IL-6 induced proliferation and apoptosis in human vascular smooth muscle cells (VSMC).Methods Recombinant plasmid pcDNA3.1-XPB and vacant vector plasmid pcDNA3.1 were transfected stably into VSMC by liposome,and these cells were incubated with IL-6 at a 100 U/ml for 48 hours.The experiments were divided into six groups:blank control group; pcDNA3.1 group; pcDNA3.1-XPB group;IL-6 group; IL-6 + pcDNA3.1 group; IL-6 + pcDNA3.1-XPB group.The expression levels of XPB,Bcl-2,Bax and wild type p53 (wt-p53) were detected through reverse transcription polymerase chain reaction (RT-PCR) and Western blotting.The cell survival,cell cycle and apoptosis were examined with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry,respectively.Results The transfection of pcDNA3.1-XPB increased the expression of XPB,Bax and wt-p53 (P＜0.05 or P＜0.01),decreased the expression of Bcl-2 (P＜0.05 or P＜0.01),and reduced the IL-6 induced effects on decreasing the expression of Bax and wt-p53 and increasing the expression of Bcl-2(P＜0.05 or P＜0.01).The over expression of XPB inhibited the cell growth(q=2.95,P＜ 0.05),and reduced the positive effects of IL-6 on VSMC growth ( 102.6 +6.2) ％ vs.(124.5 + 7.9) ％,q=3.49,P＜0.05.The over expression of XPB increased the apoptosis rate of VSMC(P＜0.01 ) and the cell amounts of G0/G1 phase (q=2.99,P＜ 0.05),decreased the cell amounts of S phase(q=3.05,P＜0.05),and reduced the IL-6 induced effects on decreasing the apoptosis rate of VSMC(5.9±2.1)％ vs.(0.3±0.1)％,q=7.53,P＜0.01; the cell amounts of G0/G1 phase(70.9±6.7) ％ vs.(54.8±2.9) ％,q=6.91,P＜0.01 ;and on increasing the cell amounts of Sphase(20.2+3.6)％ vs.(36.4+7.2)％,q=8.54,P＜0.01.Conclusions XPB gene could inhibit VSMC proliferation,promote VSMC apoptosis,and reduce the effects that IL-6 promotes VSMC proliferation and inhibits VSMC apoptosis.Therefore,XPB gene is likely to be potential molecular target for treatment of atherosclerosis.

Objective To investigate whether thalidomide inhibits the over expression of type I collagen in pulmonary fibrosis rats via inhibiting the JNK signaling pathway,thereby reducing bleomycin induced pulmonary interstitial fibrosis in rats.Methods 90 healthy male SD rats were randomly divided into normal control group (group N),model group (group M),thalidomide group (group T),SP600125 group (group SP) and thalidomide+SP600125 group (group T+SP).The pulmonary fibrosis models were prepared via intratracheal injection of 5mg/kg bleomycin,and rats in groups were given corresponding drugs from the first day after preparing model.Rats were randomly sacrificed at 7,14 and 28 days after treatment.The degree of pulmonary alveolitis and fibrosis was evaluated by H&E and trichrome masson stainings.The level of hydroxyproline in the lung tissue was detected by applying alkaline hydrolysis technique,and expression levels of p-JNK and type I collagen were tested by Western bloting for protein expression and real-time polymerase chain reaction (RT-PCR) for mRNA expression.Results In group M,alveolitis was the most serious on day 7; a marked pulmonary fibrosis formed on day 28; the level of hydroxyproline also peaked on day 28,and the contents of p-JNK and type I collagen were higher than in group N(F=277.87,472.51,both P＜ 0.01).Group T,SP and T+SP showed mild alveolitis and fibrosis at all time points,and their levels of hydroxyproline,p-JNK and type I collagen were remarkably decreased as compared with group M (F=14.77,61.59,101.73,all P＜0.01;F=10.33、79.12、57.48,all P＜0.01).No significant difference in p JNK was found between group SP and group T+SP.Conclusions Thalidomide may inhibit the over expression of type I collagen in pulmonary fibrosis rats via inhibiting the JNK signaling pathway,thereby reducing bleomycin induced pulmonary interstitial fibrosis in rats.

Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.

Objective:To preliminarily study the characteristics of ventricular synchronicity in patients of idiopathic pulmonary arterial hypertension (IPAH) with different heart function.
Methods:Our research included in 2 groups:IPAH group, n=40 patients intermittently collected in our hospital from 2010-10 to 2014-12 and Control group, n=23 normal subjects from the same period of time. Tissue Doppler echocardiography was conducted to compare interventricular systolic and diastolic time to peak (Ts and Te), their differences (Te-Ts) and (Te-Ts) of left ventricle (LV), right ventricle (RV), interventricular septum (IVS) between 2 groups.
Results:Ts and Te of RV were longer than Ts and Te of LV and IVS in both groups, P<0.05. Compared with Control group, by heart function reducing, IPAH group showed different Ts and Te of LV and IVS, P<0.05 and Ts was gradually prolonged, Te was gradually shorter;while Te of RV became longer firs tand then became shorter thereafter. In IPAH group, the patients with heart function grade IV had the longer Ts of LV and IVS than those with heart functionII, III;the patients with heart function IV had the shorter Te of LV, RV and IVS than those with heart function II, III, all P<0.05. In IPAH group with different heart function, Te between RV-IVS and RV-LV were different from Control group, all P<0.05;Te between LV-IVS in heart function IV patients had the largest difference from Control group, P<0.05. Compared with Control group, IPAH group had gradually decreased Te-Ts of LV and IVS by heart function reducing asgrade II>III>IV, all differences had statistic meaning.
Conclusion:IPAH patients with different right heart function may have interventricular systolic and diastolic de-synchronicity;by reduced heart function, interventricular Te-Ts could be specifically shortened.

Objective To study the diagnosis, treatment and prognosis of the rupture of breast implant. Methods Physical examination, X-ray fluoroscopy, high frequency and color the Doppler ultrasonography were helpful to make a clear diagnosis, in exploring and removing the fiber envelope by inframammary or periareolar incision, repairing the anatomy structure of the breasts and performing augmentation mammaplasty again. Results In 32 patients, the breast shape was basically symmetrical, most of the incisions were healed by first intention except 2 cases with complication of subcutaneous fluid collection. A satisfactory therapeautic result could be achieved by taking out the breast implant which was rupture, exploring and removing the fiber envelope, and extending the lacuna. Conclusion A satisfactory therapeautic result can be achieved by extending the lacuna.

Objective To investigate the inhibitory effect and mechanism of rosiglitazone on chemokines secretion in renal tubular epithelial cells (HK-2) stimulated by lipopolysaccharide (LPS). Methods Cells were divided into four groups: control (CON), LPS (1 mg/L),rosiglitazone (10 μmol/L), rosiglitazone (10 μmol/L) +LPS (1 mg/L). MCP-1 and IL-8 expression was measured using real time PCR and ELISA. PPARγ was knockdown by RNAi to investigate whether the inhibitory effect of rosiglitazone was PPARγ-dependent or -independent. The NF-κB in nucleus was detected by Western blotting. The DNA binding activity of NF-κB was determined by electrophoretic mobility shift assay. Results Compared with CON group, the expressions of IL-8 and MCP-1 were increased by (4.30±0.45) and (4.80±1.29) times in mRNA level, (1.39±0.18)and (2.11 ±0.47) times in protein level, respectively, in LPS-stimulated HK-2 cells (P＜0.05).Application of rosiglitazone followed by LPS significantly reduced IL-8 and MCP-1 secretion compared with LPS group (decreasing by 66.37% and 71.88% in mRNA levels, while 41.68% and 47.87% in protein levels) (P＜0.05). In pcDNATM 6.2-GW/EmGFP-miPPARγ transfected cells, IL-8and MCP-1 only were decreased by 18.16% and 16.83% in mRNA level, while 11.39% and 11.86%% in protein level in rosiglitazone pretreated group, showing no significant difference compared with LPS group. Rosiglitazone did not block NF-κB nuclear translocation while significantly inhibiting the DNA binding activity of NF-κB. Conclusions Rosiglitazone inhibits the expressions of MCP-1 and IL-8 via a PPARγ-dependent mechanism in HK-2 cells, resulting from inhibition the DNA binding activity of NF-κB.