In this study, the effects of apollon antisense oligodeoxynucleotide (ASODN) on the proliferation and apoptosis of human Lovo cells in vitro were investigated. Apollon ASODN was incubated with human colorectal Lovo cells for 48 h, the proliferation inhibition and the clone forming rates were detected by WST method and clone formation assay, respectively. The expression of apollon mRNA was analyzed by real time fluorescent quantitative reverse transcription polymerase chain reaction. The percentage of apoptotic cells and cell cycle distribution were determined by flow cytometry. The morphology of apoptotic cells was examined by fluorescence microscope. Lovo cells incubated with apollon ASODN combined with 5-fluorouracil (5-FU), cisplatin (DDP) or epirubicin (EPI) of different concentrations, cell proliferation inhibition rates were detected with WST method and IC50 was calculated. It was found that ASODN targeting apollon gene could all suppress the growth of Lovo cells and induce apoptosis of these cells significantly (P < 0.05). After Lovo cells treated with apollon ASODN for 48 hours, the expression of the apollon mRNA level was suppressed significantly. And a marked concentration-dependent decline of cell proliferation and clone forming, increasing of cell apoptosis levels were observed. The percentage of G0/G1 phage cells was abated and that of S phage cells was increased and the Lovo cells arrested at S phage of the cell cycle detected with flow cytometry. Many Lovo cells stained with Hoechst 33258 exhibited apoptotic morphology such as cell shrinkage, nuclear condensation and nuclear fragmentation. Cell proliferation inhibition was detected and their chemo-therapeutic effects of 5-FU, DDP and EPI on Lovo cells combined with apollon ASODN (0.08 micromol x L(-1)) were enhanced independently compared with single 5-FU, DDP and EPI groups, and the sensitivity enhanced about 2.58, 4.47, and 5.33 times respectively. It can be concluded that ASODN targeting apollon can suppress the expression of apollon mRNA, and inhibit the proliferation, induce apoptosis, arrest cell cycle at S phase of colorectal cancer Lovo cells in vitro and enhance the chemo-sensitivity to 5-FU, DDP and EPI.

OBJECTIVE:To study the absorption features of Silymarin enteric coated-polyllactic-co-glycolic acid (PLGA) nanoparticles in rat in situ intestine perfusion model and colonic adenoma Caco-2 cell model. METHODS:HPLC method was used to determine the content of silymarin. The absorption rate constant(Ka)and apparent absorption coefficient(Kapp)of Silymarin sus-pension,Silymarin PLGA nanoparticles and Silymarin enteric coated-PLGA nanoparticles were investigated in duodenum,jejunum, ileum and colon of rat in situ intestine perfusion model;the apparent permeability coefficient (Papp) of those drugs containing low-concentration,medium-concentration and high-concentration(20,40,60 μg/mL)of silymarin in Caco-2 cell model were also investigated. RESULTS:Compared with Silymarin suspension,Ka and Kapp of Silymarin PLGA nanoparticles and Silymarin enteric coated-PLGA nanoparticles were all increased in duodenum,jejunum,ileum and colon(P<0.05);compared with the correspond-ing concentration Silymarin suspension,two-way Papp of Silymarin PLGA nanoparticles and Silymarin enteric coated-PLGA nanopar-ticles containing low-concentration,medium-concentration and high-concentration of silymarin were all increased in Caco-2 cell model (P<0.05);there was no statistical significance between Silymarin PLGA nanoparticles and Silymarin enteric coated-PLGA nanoparticles (P>0.05). CONCLUSIONS:Silymarin enteric coated-PLGA nanoparticles can effectively increase the intestinal ab-sorption,cellular uptake and transmembrane transport rate of silymarin.

Objective To investigate the clinical value of transthyretin (TTR) from patients with early rheumatoid arthritis (ERA). Methods 58 patients with ERA , 34 patients with later RA (LRA) and 34 healthy control (HC) were included in the research. TTR was analyzed by ELISA, whose variance was analyzed. TTR density, disease activity score28 (DAS28) score and rheumatoid factor (RF) were tested, and their correlation with TTR was analyzed. Results Serum level of TTR with ERA significantly increased compared with that with LRA and HC (P < 0.05), no statistical significance with LRA group and HC. TTR level was no correlation with the number of swelling and tender joints, disease activity score 28, RF, ESR, CRP, anti-cyclic citrylinated peptide antibody and anti-keratin antibodies, hemoglobin, thrombocyte and albumin. Conclusion Serum level of TTR significantly increased with ERA patients, contributing to early diagnosis for RA.

Objective To evaluate the refinement process for extracting Herba Artemisiae Scopariae(HAS) with macroporous resin.Methods With reservation rate of main peak in the fingerprint as the index,the refinement process for extract of HAS was evaluated.The safety of oral administration of HAS extract before and after refinement process was evaluated by detecting serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST),observing the hepatic histopathological changes in mice model of CCl4-induced acute hepatic injury,and by mice acute toxicity experiment.Results The fingerprint of HAS extract before and after treatment was similar,the difference of their hepatoprotective effect was insignificant,and there was no obvious acute toxic reaction of HAS extract before and after refinement.Conclusion This experiment verified the rationality of the refinement process for extracting Herba Artemisiae Scopariae with macroporous resin by the method of pharmacodynamics combining with HPLC fingerprint.

AIM: To study the influencing factors of static adsorpting process of extract of Herba Artemisiae scopariae with macroporous resin,and determine the refinement process. METHODS: The absorbances and the adsorption quantity of caffeotannic acid were used as indexes,the adsorption effect of 9 kinds of macroporous resin and static adsorption curve of F resin;density and pH of the extract liquor were investigated by use of factorial experiment;HPLC fingerprint of extract of Herba Artemisiae scopariae were evaluated the adsorption effect.(RESULTS:) The adsorpting effect were different among types of macroporous resin,and the adsorpting equilibrium time was 6 h in the use of F resin;Density and pH of the extract liquor are important factors of adsorption;The experiment indicated that decreased liquor by this process were important factors of adsorption;The experiment indicated that this process decreased the yield of extract by 2.5%. CONCLUSION: The result can offer information about the determination of refinement process of extract of Herba Artemisiae scopariae with macroporous resin.

AIM: To study the kinetic adsorption process in the use of the extraction of Herba Artemisiae scopariae. METHODS: In combination with caffeotannic acid and the absorbance, HPLC fingerprint of extract of Herba Artemisae scopariae was adopted as marker, to investigate the kinesis adsorb and elution process, the to determine the max quantity of physic liquor and the type and quantity of elution solvent. RESULTS: The max adsorption quantity of caffeotannic acid that could adsorb was 20.9 mg/g dry resin, and the elution solvent determined was five-fold column volume of 80% alcohol; The repeated experiment indicated that this process decressed to 2.5% yield of extract and meanwhile it retained the whole components of the HPLC fingerprint effectly. CONCLUSION: The kinetic adsorption can enrich the active components in Herba Artemisiae scopariae effectively.

Objective To investigate the effects of siRNAs targeted protein kinase C ?(PKC?) on the proliferation and apoptosis of A549 cell line.Methods Six PKC? siRNAs were designed and chemical synthesized. Candidate PKC? siRNA was transfected into A549 cells,PKC? mRNA level was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Then,the effects of more effective PKC? siRNAs on A549 were further investigated by using clone formation assay,Hoechst 33258 staining,propidium iodide (PI) staining,Annexin V-FITC and PI dual staining and western blot. Results Six candidate PKC? siRNAs were designed and named as No.1,No.2,No.3,No.4,No.5 and No.6 PKC? siRNA. Compared with controls,PKC? mRNA levels were all significantly downregulated and the cell proliferation was inhibited in A549 cells treated with candidate PKC? siRNAs except No.5 (P

Deciphering the metabolites of multiple components in herbal medicine has far-reaching significance for revealing pharmacodynamic ingredients.However,most chemical components of herbal medicine are secondary metabolites with low content whose in vivo metabolites are close to trace amounts,making it difficult to achieve comprehensive detection and identification.In this paper,an efficient strategy was proposed:herb-derived metabolites were predicted according to the structural characteristics and metabolic reactions of chemical constituents in Corydalis Rhizoma and chemical structure screening tables for metabolites were conducted.The fragmentation patterns were summarized from represen-tative standards combining with specific cleavage behaviors to deduce structures of metabolites.Ion abundance plays an important role in compound identification,and high ion abundance can improve identification accuracy.The types of metabolites in different biological samples were very similar,but their ion abundance might be different.Therefore,for trace metabolites in biological samples,we used the following two methods to process:metabolites of high dose herbal extract were analyzed to char-acterize those of clinical dose herbal extracts in the same biological samples;cross-mapping of different biological samples was applied to identify trace metabolites based on the fact that a metabolite has different ion abundance in different biological samples.Compared with not using this strategy,44 more metabolites of clinical dose herbal extract were detected.This study improved the depth,breadth,and accuracy of current methods for herb-derived metabolites characterization.

Cardiac Catheterization ; Ductus Arteriosus, Patent ; therapy ; Heart Septal Defects, Atrial ; therapy ; Humans ; Printing, Three-Dimensional ; Vena Cava, Inferior

Purpose: NUF2 has been implicated in multiple cancers recently, suggesting NUF2 may play a role in the common tumorigenesis process. In this study, we aim to perform comprehensive meta-analysis of NUF2 expression in the cancer types included in the Cancer Genome Atlas (TCGA). Materials and Methods: RNA-sequencing data in 31 cancer types in the TCGA data and 11 independent datasets were used to examine NUF2 expression. Silencing NUF2 using targeting shRNAs in hepatocellular carcinoma (HCC) cell lines was used to evaluate NUF2’s role in HCC in vitro and in vivo. Results: NUF2 up-regulation is significantly observed in 23 out of the 31 cancer types in the TCGA datasets and validated in 13 major cancer types using 11 independent datasets. NUF2 overexpression was clinically important as high NUF2 was significantly associated with tumor stages in eight different cancers. High NUF2 was also associated with significantly poorer patient overall survival and disease-free survival in eight and six cancers, respectively. We proceeded to validate NUF2 overexpression and its negative association with overall survival at the protein level in an independent cohort of 40 HCC patients. Compared to the non-targeting controls, NUF2 knockdown cells showed significantly reduced ability to grow, migrate into a scratch wound and invade the 8 μm porous membrane in vitro. Moreover, NUF2 knockdown cells also formed significantly smaller tumors than control cells in mouse xenograft assays in vivo. Conclusion NUF2 up-regulation is a common feature of many cancers. The prognostic potential and functional impact of NUF2 up-regulation warrant further studies.