Objective Response surface methodology ( RSM ) was applied to optimize the ultrasonic extraction conditions for flavonoids from Coreopsis tinctoria Nutt. Methods The influence factors of ultrasonic extraction were evaluated using the Box-Behnken central component experiments and analyzed by RSM. Results The optimum extraction conditions were confirmed as follows:extraction time 30. 0 min, ratio of liquid to solid 21∶1, concentration of ethanol 60%. The yield of flavonoids under this condition was (4.65±0.036)% (n=3). Conclusion The flavonoids could be extracted with stability and higher yield from Coreopsis tinctoria Nutt under optimized conditions.

ObjectiveTo study the expression of microRNA-21 ( miR-21 )in serum of patient with diffuse large B cell lymphoma (DLBCL) and DLBCL cell lines and validate the significance of miR-21 in early diagnosis,genotyping and prognosis estimates of DLBCL.MethodsmiR-21 expression were detected by fluorescent quantity polymerase chain reaction (FQ-PCR)in 9 lymphoma cell lines (OCI-Ly1,OCI-Ly3,OCI-Ly4,OCI-Ly7,OCI-Ly8,OCI-Ly10,OCI-Ly18,OCI-Ly19 and HBL),the serum from DLBCL patients (n =62) and health controls (n =50 ).Kaplan-Meier survival analysis was carried out during the relapsefree survival period of DLBCL patients to explore the relationship between the prognosis and microRNA expression level.ResultsReal time FQ-PCR result indicated that miR-21 expression was higher in DLBCL cell lines than that in normal B cells (BC).miR-21 expression in normal B cell and 9 DLBCL cell lines separately were 1.04 ± 0.02,2.30 ± 0.35,237.97 ± 56.19,5.27 ± 0.83,3.40 ± 0.30,11.22 ± 2.70,133.55 ± 16.78,6.63 ±0.24,4.91 ±0.37 and 81.59 ±6.64.Compared with BC,the expression of miR-21 were higher in all 9 DLBCL cell lines ( t =7.3,13.7,21.0,6.2,8.8,13.6,6.5,39.5,18.1 ;P ＜ 0.01 ).miR-21 expression segregates with specific molecular subgroups of DLBCL The expression was higher in the ABC type cell lines (OCI-Ly3,OCI-Ly10,HBL) than GCB type cell lines (OCI-Ly1,OCI-Ly4,OCI-Ly7,OCI-Ly8,OCI-Ly18,OCI-Ly19;t =11.18,P ＜ 0.01 ).Consistent with the cell line models,miR-21 expression levels were higher in serum from DLBCL patients [21.38 (10.26-45.21 )] than from controls [1.87 ( 1.05-3.97 ),U =168,P =0.000],and the levels were higher in DLBCL cases with an ABC-type [28.68 ( 14.92-98.44 )] than those in GCB-type [18.30 ( 7.32-33.46 ),U =336,P =0.043].MiR-21 expression levels were different in sera from different clinical stage DLBCL patients.The miR-21 level in serum of patients with subgroup ABC and subgroup GCB in stage Ⅰ and Ⅱ were 47.49( 25.65-295.41 ) and 24.74( 16.08-50.38) respectively and in stage Ⅲ and Ⅳ were 16.66 ( 5.35-44.30 ) and 11.96 ( 4.10-21.05) respectively.The levels were higher in DLBCL cases withⅠ -Ⅱ stage than those with Ⅲ-Ⅳ stage (U =62,P =0.013 in GCB type; U =53,P =0.014 in ABC type).Moreover,compare with relapse-free survival in DLBCL patients,high miR-21 expression was associated with well prognosis ( U =259,P =0.035).ConclusionsMiR-21 is high expression in DLBCL cell lines and DLBCL patients serum.miR-21 level in sera from DLBCL patients is associated with clinical stage,molecular subgroup and prognosis estimates.MiR-21 may serve as a new biomarker to early detection,genotyping and prognosis estimates of DLBCL.

Objective: To explore the relationship between intra-operative blood product transfusion and post-operative prognosis in children with pediatric open heart surgery by cardiopulmonary bypass (CPB).
Methods: A total of 1028 consecutive pediatric patients received the open heart surgery by CPB in our hospital were retrospectively studied. Uni- and multivariate Logistic regression analysis were conducted to analyze the relationship between intra-operative blood product transfusion and post-operative in-hospital death and complication rates.
Results: There were 45/1028 (4.4%) of patients died, 143 (13.9%) with low cardiac output syndrome, 43 (4.2%) received dialysis, 26 (2.5%) with sustained pulmonary failure, 17 (1.7%) with infection, and 28 (2.7%) developed neurologic complications. Multivariate Logistic regression analysis indicated that duration and the lowest temperature of CPB, 24-hour post-operative pediatric risk of mortality (PRISM-III) score and intra-operative transfusion amount of RBC > median (20.3 ml/kg) were the independent risk factors for post-operative in-hospital death. Congenital heart disease (CHD) with pre-operative cyanosis, duration of CPB, 24-hour post-operative PRISM-III score, 6-hour post-operative mediastinal drain loss and intra-operative transfusion amount of RBC > 20.3 ml/kg were the independent risk factors for post-operative complication occurrence.
Conclusion: Large volume of RBC transfusion in the open heart pediatric surgery with CPB may increase the risk of post-operative in-hospital death and complication occurrence, reducing RBC transfusion might be improving the post-operative prognosis in CHD patients.

We investigated the roles of the crude antigen(CA) of Clonorchis sinensis and excretory secretory products (ESPs) in the polarization of Th1 and Th2 cells.Bone marrow-derived cells were generated from BALB/c mice and isolated into immature DCs;immature DCs were then treated with either CA (CA stimulated group),ESPs (ESPs stimulated group),LPS (positive control group) or PBS (negative control group) for 24 hours.Then the CD4+T cells were isolated from mouse spleen by using anti mouse-CD4 Microbeads,and further cocultured with stimulated DCs for another 72 hours.The purities of DCs and CD4+ T cells were evaluated by flow cytometry and the expressing levels of T-bet mRNA and GATA-3 mRNA were detected by real-time PCR.ELISA was used to detect the levels of IFN-γ and IL-4 cytokines in the supernatant.mRNA levels of T-bet and GATA-3 in the ESPs group were higher than those in PBS-stimulated group (P＜0.05).The concentrations of IFN-γ and IL-4 cytokines in the culture were increased in the ESPs group,compared with PBS stimulated group(P＜0.05).IFN-γ but not IL-4 was increased in CA group (P＜0.05).The results implied that CA might play a role in Th1 type immune response,and ESPs likely play roles in both Th1 and Th2 immune responses.

Objective To provide a comprehensive reference index for different mouse models of herpes simplex virus 1 (HSV-1) infection by investigating the related clinical manifestations, pathological features and characteristics of viral distribution in tissues and organs of BALB/c mice infected with different HSV-1 strains by using different strategies.Methods Acute infection models were established by challenging BALB/c mice at age three or six weeks with HSV-1 17+ and McKrae strains via intranasal and corneal administrations.Correspondingly, chronic infection models were established with BALB/c mice through subcutaneous and foot pad injections.Results Although all experimental mice showed trichiasis and roachback, there were differences in weight and fatality rate among different groups.Results of the quantitative PCR detection indicated that the proliferation of HSV-1 in the nervous tissues (brain, spinal cord, trigeminal ganglion) varied among different groups.The pathological examination indicated that in the acute infection groups, significant pathological changes only occurred in the brain tissues, while in the chronic infection groups, pathological injuries only occurred in the trigeminal ganglia.Although a key index latency-associated transcript (LAT) was not detected in the trigeminal nerve tissues of mice in the chronic infection groups, co-culturing the tissues with Vero cells resulted in infectious lesions in the cells.Conclusion This study indicates that there are significant differences in weight and fatality rate among different BALB/c mouse models of HSV-1 infection.Varied replication dynamics of HSV-1 were observed in different tissues or organs of the BALB/c mice in different groups.Therefore, different indexes should be adopted to evaluate different HSV-1 infection models.

Objective To investigate the effects of cholesterol-lowering agents on the proliferation, stemness characters, migration, invasion, and neutrophil extracellular traps formation (NETs) formation in liver cancer cells. Methods ASPP2 or HMGCR gene was knocked down in mouse liver cancer cell Hepa1-6 to establish cells with high or low cholesterol, respectively. Simvastatin and berberine were used to reduce cholesterol synthesis. CCK-8 and plate cloning assays were conducted to detect the proliferation ability of liver cancer cells. Sphere formation assay and qRT-PCR were used to analyze the stemness character and expression of related genes. Wound-healing assay and Transwell assay were used to analyze the ability of cell migration and invasion. Immunofluorescence staining was carried out to analyze the effect of lipid-lowering agent on NETs formation. Results Cholesterol-lowering agents significantly inhibited the proliferation and stemness-related gene expression of Hepa1-6 cells (P < 0.001), significantly inhibited the migration and invasion of Hepa1-6 cells (P < 0.001), and significantly inhibited the neutrophil-induced invasion and formation of NETs (P < 0.001). Conclusion Cholesterol-lowering agents suppress the proliferation and invasion via inhibiting the stemness characters and NETs formation in liver cancer cells. It is a potential strategy for the treatment of liver cancer metastasis.

In order to study effects of ginseng on the metabolism of drug belong to CYP3A4 substrate, screening of pregnane X receptor activation from ginsenosides was performed by reporter assay. Based on PXR-CYP3A stable translation cell lines, 13 ginsenosides were screened for pregnane X receptor activation by reporter assays, and RIF as the positive control. The effect of ginsenosides Rg1 onCYP3A4 mRNA expression was also investigated by RT-PCR. The PXR-CYP3A stable translation cell lines had good response to RIF, and the EC50 is 2.51 micro mol x L(-1). When the condition of final concentration was 10 micromol x L(-1), ginsenoside F2 and protopanaxatriol had moderate inductive effects on PXR. Panaxotriol, Rg2, pseudoginsenoside F11, Rg1, ginsenoside and Rb3 had inhibitory effects on PXR. Ginsenoside Rf1, Rg3, Rh2 and protopanaxdiol had no obvious effects on PXR. Rg1 down-regulated CYP3A4 mRNA expression in a concentration-dependent manner. Activation of pregnane X receptor by ginsenosides may influence the metabolism of drug belong to CYP3A4 substrate, and cause ginseng-drug interactions.

We analyzed the whole genome coding sequence of Volvariella volvacea to study the pattern utilization of codons by Codon W 1.4.2. As results, 24 optimal codons were identified. Moreover, the frequency of codons usage was calculated by CUSP program. We compared the frequency of codons usage of V. volvacea with other organisms including 6 modal value species (Homo sapiens, Saccharomys cerevisiae, Arabidopsis thalian, Mus musculus, Danio rerio and Drosophila melanogaster) and 4 edible fungi (Coprinopsis cinerea, Agaricus bisporus, Lentinula edodes and Pleurotus ostreatus). We found that there were less differences in 3 edible fungi (excluding Pleurotus ostreatus) than 6 modal value species, comparing with the frequency of codons usage of V. volvacea. With software SPSS16.0, cluster analysis which showed differences in the size of codon bias, reflects the evolutionary relationships between species, which can be used as a reference of evolutionary relationships of species. This was the first time for analysis the codon preference among the whole coding sequences of edible fungi, serving as theoretical basis to apply genetic engineering of V. volvacea.
Agaricales ; genetics ; Animals ; Arabidopsis ; genetics ; Cluster Analysis ; Codon ; DNA, Fungal ; genetics ; Drosophila melanogaster ; genetics ; Humans ; Mice ; Saccharomyces cerevisiae ; genetics ; Software ; Volvariella ; genetics ; Zebrafish ; genetics

Objective To explore heterogeneous subtypes of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation and the characteristics differences of different types of children after liver transplantation. Methods Seven hundred and forty-one children who underwent living-related liver transplantation were enrolled. The self-designed general information questionnaire, Chinese version of 5-Item World Health Organization Well-Being Index (WHO-5) and the parent-report version of the Strengths and Difficulties Questionnaire (SDQ) were filled out by their guardians. The scores of five dimensions of SDQ were used as the manifest variables of the model. The classification model of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation was constructed by latent profile analysis. The latent categories of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation were analyzed. The influencing factors of latent categories were analyzed by univariate analysis and logistic regression model. Results There were three latent categories of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation, including peer communication problem group (n=302), psychological and behavioral adaptation group (n=145) and psychological and behavioral adjustment difficulty group (n=294). The first two groups were merged into the psychological and behavioral health group (n=447), which had significant differences in the five dimensions and the total score of difficulties of SDQ compared with the psychological and behavioral adjustment difficulty group (n=294) (all P<0.001). Logistic regression analysis showed that age≤5 years old, primary disease of non-cholestatic liver disease, stem family were the risk factors for psychological and behavioral adjustment difficulties in pediatric recipients after liver transplantation. Female gender, high education levels of parents and high WHO-5 score of guardians were the protective factors for psychological and behavioral adjustment difficulties in pediatric recipients after liver transplantation (all P<0.05). Conclusions The psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation are heterogeneous. Medical staff should pay extensive attention to different characteristics of pediatric recipients after liver transplantation with different psychological and behavioral adaptation categories and adopt targeted screening and intervention strategies, aiming to improve psychological and behavioral adaptation outcomes of pediatric recipients after liver transplantation.

Objective To assess alternations in left ventricular( LV) torsion parameters in healthy subjects and patients with premature ventricular complexes ( PVCs) from the right ventricular outflow tract (RVOT) .Methods ThirtypatientswithPVCsfromRVOTand31healthysubjectswereincluded.Two-dimensional speckle tracking imaging( 2D-STI) was applied to evaluate LV torsion parameters ,including LV rotational degrees in basal and apical levels respectively ,LV total torsional degrees ,and time to peak rotational and torsional degrees . All values of patients with PVCs were recorded during both sinus beats ( PVC-S) and premature ventricular beats ( PVC-V) . Results No significant difference was observed in left ventricular ejection fraction(LVEF) between PVC-S group and control subjects( P > 0 .05) ,while LV rotational degrees in apical levels[( 8 .47 ± 3 .54)° vs (9 .50 ± 3 .21)° , P = 0 .042] and LV total torsional degrees [ (11 .25 ± 6 .31)° vs (14 .00 ± 4 .07)° , P =0 .046] were significantly reduced in PVC-S group . In addition to the reduction of LV rotational degrees in apical levels[( 3 .93 ± 7 .23)° vs (9 .50 ± 3 .21)° , P =0 .000] and LV total torsional degrees[ ( 4 .35 ± 9 .62)° vs (14 .00 ± 4 .07)° , P =0 .000] ,lower apical levels[ ( -0 .57 ± 4 .44)° vs ( -5 .26 ± 3 .84)° , P =0 .000] and advanced LV rotational degrees [ ( 40 .5 ± 18 .6)％vs (48 .0 ± 9 .1)％ , P =0 .05] in basal levels were observed in PVC-V group in comparison with the control subjects . Compared with the PVC-S group ,PVC-V group showed lower LV rotational degrees in basal levels [ ( -0 .57 ± 4 .44)° vs ( -4 .57 ± 4 .57)° , P = 0 .000] and advanced time to peak rotational degrees [ (40 .5 ± 18 .6)％ vs (48 .1 ± 12 .6)％ , P =0 .018] ,as well as advanced time to peak and lower LV total torsionaldegrees[(39.3±15.4)％ vs(46.7±13.8)％ ,P =0.007 ;(4.35±9.62)°vs(11.25±6.31)°,P=0 .001] .Conclusions As to RVOT-PVC patients ,LV myocardial torsional motion has changed in PVC-S mainly manifested as a decrease of rotation degrees in apical levels even if the LVEF is still in the normal range . During PVC-V the rotation and twist degree is further reduced ,and the time sequence altered , accompanied with significantly decreased LVEF .