The region of about 4.0 kb upstream of Spodoptera litura nucleopolyhedrovirus (SpltNPV) chiA gene was sequenced, in which six open reading frames(ORF1～6) were found. These ORFs are 156, 297, 540, 369, 1281, and 228 nucleotides long, encoding the proteins of 51, 98, 179, 122, 426, and 75 amino acids with the molecular weight of 6.15 kD, 11.46 kD, 21.70 kD, 14.69 kD, 47.59 kD, and 9.09 kD respectively. One early promoter motif CAGT in ORF1 and ORF3, two early promoter motifs CAGT in ORF2, one late promoter motif TAAG in ORF4 and two late promoter motifs TAAG in ORF5 were found in 5′noncoding regions of these ORFs. The polyadenylation signals, AATAAA, are located downstream of the translation stop codon of ORF1, ORF4 and ORF5. ORF4 is the homologous gene of AcMNPV ORF53, BmNPV ORF42, OpMNPV ORF56 and LdMNPV ORF54. Compared with all genes from baculoviruses, ORF1, ORF2 and ORF6 have no homologous genes. It is suggested that ORF1, ORF2 and ORF6 may be three novel baculovirus genes.

A series of cycloberberine derivatives were designed, synthesized and evaluated for their anti-cancer activities in vitro. Among these analogs, compounds 6c, 6e and 6g showed strong inhibition on human HepG2 cells. They afforded a potent effect against DOX-resistant MCF-7 breast cells as well. The primary mechanism showed that cell cycle was blocked at G2/M phase of HepG2 cells treated with 6g using flow cytometry assay. It significantly inhibited the activity of DNA Top I at the concentration of 0.1 mg mL-1. Our results provided a basis for the development of this kind of compounds as novel anti-cancer agents.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Berberine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type I ; metabolism ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Hep G2 Cells ; Humans ; MCF-7 Cells ; Molecular Structure ; Structure-Activity Relationship

Objective:To investigate the expression of Fas、FasL and the apoptosis of liver cancer cell line HepG2 transfected with LIGHT and IFN-? gene mediated by Cationic liposome.Methods:HepG2 cells were divided into two groups(the solo transfection of LIGHT gene and the combined transfection of LIGHT and IFN-? genes) and the control groups(no transfection).HepG2 cells were cellected at 12h,24h and 48h after transfection.The apoptosis of HepG2 cells and the expression of Fas and FasL of the HepG2 cells were investigated with flow cytometry.Results:After transfection,the apoptosis of HepG2 cells increased,and the apoptosis of combined transfection group was higher than the solo transfection of LIGHT(P

OBJECTIVE: To study the expression levels of glial cell line-derived neurotrophic factor family receptor α-1 (GFRα1) and enhancer of zeste homolog 2 (EZH2) in the intestinal tissue of children with Hirschsprung's disease (HSCR), as well as the role of EZH2 in the regulation of GFRα1 gene expression and the pathogenesis of HSCR. METHODS: The samples of colon tissue with spasm from 24 children with HSCR after radical treatment of HSCR were selected as the experimental group, and the samples of necrotized colon tissue from 18 children with neonatal necrotizing enterocolitis after surgical resection were selected as the control group. Real-time PCR and Western blot were used to measure the expression levels of GFRα1 and EZH2 in colon tissue in both groups. Human neuroblastoma SH-SY5Y cells were divided into an EZH2 over-expression group and a negative control group. The cells in the EZH2 over-expression group were transfected with pCMV6-EZH2 plasmid, and those in the negative control group were transfected with pCMV6 plasmid. The expression levels of EZH2 and GFRα1 were measured after transfection. RESULTS: Compared with the control group, the experimental group had significant reductions in the mRNA and protein expression levels of GFRα1 and EZH2 in colon tissue (P<0.05), and the protein expression of EZH2 was positively correlated with that of GFRα1 (r=0.606, P=0.002). Compared with the negative control group, the EZH2 over-expression group had significant increases in the expression levels of EZH2 and GFRα1 after SH-SY5Y cells were transfected with EZH2 over-expression plasmid (P<0.05). CONCLUSIONS Low expression of EZH2 in the colon tissue of children with HSCR may be one of the causes of inadequate expression of GFRα1 and onset of HSCR.
Child ; Colon ; Enhancer of Zeste Homolog 2 Protein ; genetics ; Glial Cell Line-Derived Neurotrophic Factor Receptors ; genetics ; Hirschsprung Disease ; genetics ; Humans ; Infant, Newborn ; RNA, Messenger

OBJECTIVETo study the levels of blood lipid among Chinese adults in 2010 and analyze the epidemiological features.

METHODSFasting serum levels of total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were determined for 90 395 subjects aged 18 and over who were selected by multi stage cluster random sampling method from 162 surveillance sites of 31 provinces of China mainland in 2010. After complex weighting, the differences of levels of serum total TC, TG, HDL-C and LDL-C were analyzed in different age, gender and geographic regions.

RESULTSAfter complex weighting, the levels of serum TC in Chinese adults was (4.04 ± 1.08) mmol/L, with (3.86 ± 1.03), (4.29 ± 1.11) and (4.33 ± 1.09) mmol/L in the groups of 18 - 44, 45 - 59 and over 60 years old (F = 177.56, P < 0.01), respectively. The level of serum TC was (4.06 ± 1.09) mmol/L and (4.03 ± 1.07) mmol/L in males and females (F = 2.38, P > 0.05), respectively; (4.08 ± 1.13) mmol/L and (4.03 ± 1.06) mmol/L in urban and rural areas (F = 0.51, P > 0.05), respectively, and (4.13 ± 1.11), (4.03 ± 1.01) and (3.93 ± 1.11) mmol/L in areas of east, middle and west respectively (F = 2.56, P > 0.05). The levels of serum TG was (1.33 ± 1.28) mmol/L, with(1.26 ± 1.24), (1.48 ± 1.46) and (1.35 ± 1.05) mmol/L in the groups of 18 - 44, 45 - 59 and over 60 years old (F = 85.98, P < 0.01), respectively. The level of serum TG was (1.45 ± 1.48) mmol/L and (1.21 ± 1.01) mmol/L in males and females (F = 172.01, P < 0.01), respectively, (1.34 ± 1.26) mmol/L and (1.33 ± 1.28) mmol/L in urban and rural areas (F = 0.16, P > 0.05), respectively, and(1.30 ± 1.25), (1.35 ± 1.23) and (1.35 ± 1.36) mmol/L in areas of east, middle and west (F = 0.69, P > 0.05) respectively. The levels of serum HDL-C was (1.11 ± 0.31) mmol/L, with (1.09 ± 0.30), (1.13 ± 0.33) and (1.14 ± 0.33) mmol/L in the groups of 18 - 44, 45 - 59 and over 60 years old (F = 27.81, P < 0.01), respectively. The level of serum HDL-C was (1.08 ± 0.32) mmol/L and (1.14 ± 0.30) mmol/L in males and females (F = 178.66, P < 0.01), respectively, (1.11 ± 0.31) mmol/L and (1.11 ± 0.32) mmol/L in urban and rural areas (F = 0.09, P > 0.05), respectively, and (1.12 ± 0.32), (1.12 ± 0.31) and (1.09 ± 0.32) mmol/L in areas of east, middle and west (F = 1.06, P > 0.05) respectively. The levels of serum LDL-C was (2.27 ± 0.78) mmol/L, with (2.15 ± 0.74), (2.42 ± 0.80) and (2.46 ± 0.81) mmol/L in the groups of 18 - 44, 45 - 59 and over 60 years old (F = 129.84, P < 0.01), respectively. The level of serum LDL-C was (2.30 ± 0.78) mmol/L and (2.24 ± 0.78) mmol/L in males and females (F = 25.06, P < 0.01), respectively, (2.33 ± 0.82) mmol/L and (2.24 ± 0.76) mmol/L in urban and rural areas (F = 3.48, P > 0.05), respectively, and (2.35 ± 0.80), (2.25 ± 0.74) and (2.17 ± 0.77) mmol/L in areas of east, middle and west (F = 4.58, P < 0.05) respectively.

CONCLUSIONThe levels of serum lipid of middle aged male, female after menopause and people living in eastern region were higher than other Chinese adults. Effective measures should be taken earlier to control serum lipid levels.

Adolescent ; Adult ; Age Distribution ; Asian Continental Ancestry Group ; China ; epidemiology ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Cross-Sectional Studies ; Dyslipidemias ; epidemiology ; Female ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Sex Distribution ; Triglycerides ; blood ; Young Adult

A series of novel N-(2-arylethyl) isoquinoline derivatives were designed, synthesized and evaluated for their anti-cancer activities. Among these analogs, compound 9a exhibited the potential anti-cancer activities on HepG2 and HCT116 cells with IC50 values of 2.52 and 1.99 microg x mL(-1), respectively. Cell cycle was blocked at S phase of HepG2 cells treated with 9a by flow cytometry detection. Our results provided a basis for the development of a new series of anti-cancer candidates.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; HCT116 Cells ; Hep G2 Cells ; Humans ; Inhibitory Concentration 50 ; Isoquinolines ; chemical synthesis ; chemistry ; pharmacology ; Molecular Structure ; Structure-Activity Relationship

Metabolic profile of bile acids was used to evaluate hepatotoxicity of mice caused by ethanol extraction of Dioscorea bulbifera L. (ethanol extraction, ET) and diosbulbin B (DB), separately. Ultra-performance liquid chromatography coupled with quadrupole mass spectrometry (UPLC-MS) was applied to determine the contents of all kinds of endogenous bile acids including free bile acids, taurine conjugates and glycine conjugates. Obvious liver injuries could be observed in mice after administrated with ET and DB. Based on the analysis using principle components analysis (PCA), toxic groups could be distinguished from their control groups, which suggested that the variance of the contents of bile acids could evaluate hepatotoxicity caused by ET and DB. Meanwhile, ET and DB toxic groups were classified in the same trends comparing to control groups in the loading plot, and difference between the two toxic groups could also be observed. DB proved to be one of the toxic components in Dioscorea bulbifera L. Bile acids of tauroursodeoxycholic acid (TUDCA), taurochenodeoxycholic acid (TCDCA), taurocholic acid (TCA), taurodeoxycholic acid (TDCA), cholic acid (CA) and others proved to be important corresponds to ET and DB induced liver injury according to analysis of partial least square-discriminant analysis (PLS-DA) and the statistical analysis showed that there were significant differences between the control groups and toxic groups (P < 0.01). Furthermore, good correlation could be revealed between the foregoing bile acids and ALT, AST. It indicated that taurine conjugated bile acids as TUDCA, TCDCA, TCA and TDCA along with CA could be considered as sensitive biomarkers of ET and DB induced liver injury. This work can provide the base for the further research on the evaluation and mechanism of hepatotoxicity caused by Dioscorea bulbifera L.
Animals ; Bile Acids and Salts ; metabolism ; Chemical and Drug Induced Liver Injury ; metabolism ; Cholic Acid ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Dioscorea ; toxicity ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Heterocyclic Compounds, 4 or More Rings ; isolation & purification ; toxicity ; Least-Squares Analysis ; Male ; Mice ; Mice, Inbred ICR ; Plants, Medicinal ; toxicity ; Principal Component Analysis ; Rhizome ; toxicity ; Tandem Mass Spectrometry ; methods ; Taurochenodeoxycholic Acid ; metabolism ; Taurocholic Acid ; metabolism ; Taurodeoxycholic Acid ; metabolism

OBJECTIVETo understand the prevalence of hypercholesterolemia, high blood low density lipoprotein cholesterol, low blood high density lipoprotein cholesterol and hypertriglyceridemia among Chinese adults in 2010.

METHODSLevels of triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C) and total cholesterol (TC) were determined in fasting serum for subjects who were selected by multi-stage stratified cluster random sampling in from 162 surveillance points of 31 provinces in 2010. Prevalence of hypercholesterolemia, high blood LDL-C, low blood HDL-C and hypertriglyceridemia of 97 409 subjects was analyzed.

RESULTSAfter the complex weighting, prevalence of hypercholesterolemia in Chinese adults was 3.3%, with 2.2%, 4.7% and 4.7% in the groups of 18 - 44, 45 - 59 and over 60 years old (P < 0.01), respectively, 3.4% and 3.2% in males and females (P > 0.05), respectively, 4.2% and 2.9% in urban and rural areas (P < 0.01), 4.2%, 2.4% and 3.1% in areas of east, central and west (P < 0.01). Prevalence of high blood LDL-C was 2.1%, with 1.3%, 3.0% and 3.6% in the groups of 18 - 44, 45 - 59 and over 60 years old (P < 0.01), respectively, 2.1% in both males and females (P > 0.05), 3.0% and 1.8% in urban and rural areas (P < 0.01), and 2.9%, 1.5% and 1.8% in areas of east, central and west (P < 0.01), respectively. Prevalence of low blood HDL-C was 44.8%, with 46.9%, 42.6% and 41.2% in the groups of 18 - 44, 45 - 59 and over 60 years old (P < 0.01), 50.6% and 38.9% in males and females (P < 0.01), respectively, 45.4% and 44.6% in urban and rural areas (P < 0.05), and 43.5%, 43.3% and 48.6% in areas of east, central and west (P < 0.01). Prevalence of hypertriglyceridemia was 11.3%, with 10.1%, 14.2% and 10.8% in the groups of 18 - 44, 45 - 59 and over 60 years old (P < 0.01), respectively, 13.8% and 8.6% in males and females (P < 0.01), respectively, 12.1% and 10.9% in urban and rural areas (P < 0.01), and 11.0%, 11.7% and 11.2% in areas of east, central and west (P < 0.05).

CONCLUSIONLow blood HDL-C and hypertriglyceridemia are two major types of dyslipidemia in Chinese adults.

Adolescent ; Adult ; Age Distribution ; Asian Continental Ancestry Group ; China ; epidemiology ; Dyslipidemias ; epidemiology ; Female ; Humans ; Hypercholesterolemia ; epidemiology ; Hypertriglyceridemia ; epidemiology ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Sex Distribution ; Young Adult

OBJECTIVETo establish the tridimensional culture method for tissue-engineered skin to observe the histomorphological change in human immortal KC strain (HacaT)cocultured with xenogenic acellular dermal matrix (ADM).

METHODSThe ADM was prepared from SD rats by a modified method. HaCaTs were cultured in defined KC-serum free medium. HaCaTs in log growth phase were inoculated on ADM at the cell density of 2 x 10(5)/cm(2). They were submergedly cultured for 5 days and then changed to air-liquid phase culture for another 5 days. ADM and growth of HaCaTs on day 1 and 5 after cocultured with ADM were observed with scanning electron microscope. The histological change in ADM and HaCaTs on day 1, 5, and 10 after cocultured with ADM were examined by HE staining.

RESULTSThe gross appearance of ADM was white with smooth and soft texture, and intact collagen bundles without cellular residue. HaCaTs adhered and stretched out pseudopodia on the surface of the ADM on day 1 after combined culture, and a monolayer of cells was formed on day 5, growing into 3-6 layers of cells on day 10 with a tendency to grow into ADM.

CONCLUSIONSSD rats ADM is benefit for the adhesion of HaCaTs and the permeation of nutrient solution, from which an engineered multiple-layered human skin can be obtained within 10 days.

Animals ; Cells, Cultured ; Coculture Techniques ; Dermis ; cytology ; Humans ; Keratinocytes ; cytology ; Rats ; Rats, Sprague-Dawley ; Skin, Artificial ; Tissue Engineering ; methods

Considering that some antibacterial agents can identify the outer structure of pathogens like cell wall and/or cell membrane, we explored a self-enhanced targeted delivery strategy by which a small amount of the antibiotic molecules were modified on the surface of carriers as targeting ligands of certain bacteria while more antibiotic molecules were loaded inside the carriers, and thus has the potential to improve the drug concentration at the infection site, enhance efficacy and reduce potential toxicity. In this study, a novel targeted delivery system against methicillin-resistant Staphylococcus aureus (MRSA) pneumonia was constructed with daptomycin, a lipopeptide antibiotic, which can bind to the cell wall of S. aureus via its hydrophobic tail. Daptomycin was conjugated with N-hydroxysuccinimidyl-polyethylene glycol-1,2-distearoyl-sn-glycero-3-phosphoethanolamine to synthesize a targeting compound (Dapt-PEG-DSPE) which could be anchored on the surface of liposomes, while additional daptomycin molecules were encapsulated inside the liposomes. These daptomycin-modified, daptomycin-loaded liposomes (DPD-L[D]) showed specific binding to MRSA as detected by flow cytometry and good targeting capabilities in vivo to MRSA-infected lungs in a pneumonia model. DPD-L[D] exhibited more favorable antibacterial efficacy against MRSA than conventional PEGylated liposomal daptomycin both in vitro and in vivo. Our study demonstrates that daptomycin-modified liposomes can enhance MRSA-targeted delivery of encapsulated antibiotic, suggesting a novel drug delivery approach for existing antimicrobial agents.