OBJECTIVE To explore the protective effect of polysaccharides from Salvia Chinensis Benth. (PSSC) on lipopolysaccharide (LPS) and D- galactosamine (GalN)- provoked mouse acute liver failure (ALF) and the possible molecular mechanism. METHODS Kunming mice were randomly divided into four groups: normal control, model, model+PSSC 30 and 100 mg·kg- 1 groups. PSCC was given once a day and for a week. To establish an ALF model, mice of model and PSSC groups were ip injected with LPS 10 μg·kg-1 and GalN 700 mg·kg-1 at the end of PSSC treatment. The microscopic structure of the liver was detected by HE staining. Serum and hepatic biochemical parameters of glutamic-oxalacetic transaminase (GOT), glutamic- pyruvic transaminase (GPT), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and glutathione (GSH) were detected by colorimetric methods. The relative content of hepatic reactive oxygen species (ROS) was measured by DCFH-DA fluorescent probes. Levels of cytokines tumor necrosis factor-α (TNF-α), interleukin 1β(IL-1β) and IL- 6 in the serum and liver were detected by ELISA. Activity of caspase 3 in liver homogenates was detected by aspase 3 activity assay kit. RESULTS Compared with normal control group, in the liver of model group, hepatocytes were arrayed in disorder, cytoplasm of hepatocytes shrank, and boundaries between cells were fuzzy, the infiltration of a large number of inflammatory cells and tissue hemorrhage could be detected, pathological scores were elevated significantly (P<0.01), levels of MDA and ROS in the liver of ALF model mice were elevated to 2.2 and 4.3 times that of the normal control, respectively (P<0.01), the level of GSH decreased to 51% (P<0.01), and the activities of SOD, CAT and GSH- Px declined to 74%, 36% and 42%, respectively (P<0.01). Levels of TNF- α, IL- 1β and IL- 6 in the serum and liver of model group were increased (P<0.01), and caspase 3 activity was increased to 5.3 times that of the normal control (P<0.01). Compared with the model group, the number of surviving mice in PSSC groups increased, liver pathological scores declined (P<0.01), levels of MDA and ROS increased (P<0.01), levels of GSH, TNF-α, IL-1β and IL-6 in the liver and serum declined (P<0.01), and caspase 3 activity decreased (P<0.01). CONCLUSION PSSC is able to alleviate LPS and GalN-induced ALF in mice. Inhibition of hepatic oxidative stress, inflammatory response, and cell apoptosis is possibly implicated in the protective effect of PSSC.

OBJECTIVE To investigate the HIV infections states in tumor patients for clinical diagnosis,treatment and to prevent HIV infection in the tumor hospital.METHODS The result of HIV detection in tumor patients from Dec 2000 to Aug 2006 was analyzed by the review statistics analysis.RESULTS Totally 48 101 tumor paients were detected,and the number of tumor patients with positive HIV antibody result was 51(0.106%).Among the positive patients there were 21 cases with blood transfusion history,14 cases with blood donating experience,2 cases with both these two kinds of experiences and 14 cases without the two kinds of experiences.Their rate was separately 41.0%,27.5%,4.0% and 27.5%.Most of the HIV positive patients had no clinical synptoms.CONCLUSIONS The HIV positive rate of patients with blood transfusion or blood donating is significantly higher than the patients without these experiences.The routine detection for the HIV before the operation,blood transfusion or other traumatic detection is very necessary.

Chemical constituents from the acetone extract of twigs of Manglietia hookeri were isolated and purified by various column chromatographic methods over silica gel and sephadex LH-20, and preparative TLC. The structures of these compounds were identified on the basis of physicochemical properties and spectral analysis, including NMR and MS spectra. Six eudesmane sesquiterpenes were obtained and their structures were identified as trans-eudesmane-4, 11-diol(1), β-eudesmol(2), (-) -10-epi-5β-hydroxy-β-eudesmol (3), epi-carrisone (4), 6-hydroxy-eudesm-4(14) -ene(5) and gynurenol(6). All the compounds were isolated from this plant for the first time. Furthermore, the 13C-NMR data of compound 3 were reported for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Magnolia ; chemistry ; Molecular Structure ; Plant Stems ; chemistry ; Sesquiterpenes, Eudesmane ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization

The method was established study the influence of different herbal combination with Radix Aconiti in the traditional medical formulae on content of the aconite alkaloids, for elucidating the scientific basis of reducing the toxicity of aconite in traditional Chinese medical formulation. The samples for ESI-MS study were prepared by decocting a mixture of Radix Aconiti Lateralis Preparata ( RALP) with Radix Glycyrrhizae Preparata (RGP) , Radix Paeoniae Alba ( RPA) , Rhizoma Zingiberis (RZ) or Radix Et Rhizoma Rhei ( RERR) , separately, and extracting the residue of the above mentioned mixtures after decocting. The diester-diterpenoid alkaloids (DDAs) was lower in the herb couples of RALP-RGP, RALP-RPA, RALP-RZ and RALP-RERR, and lipo-alkaloids was increased in the herb couples of RALP-RGP, RALP-RPA and RALP-RZ. The reason of reducing toxic effect principle is that the components of RGP, RPA and RZ have ester-exchange reactions with DDAs in RALP to produce lipo-alkaloids of low toxicity in the decocting process of the herb couples. The combination of RALP-RERR can reduce the content of DDAs in decoction and residue due to the formation of water insoluble alkaloid compound.
Aconitine ; analogs & derivatives ; analysis ; chemistry ; Aconitum ; chemistry ; Alkaloids ; analysis ; chemistry ; Diterpenes ; analysis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Esterification ; Ginger ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Hot Temperature ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rheum ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods

To produce membrane-permeable human epidermal growth factor (hEGF), a pPTD-hEGF prokaryocyte expression vector was constructed and transformed into E. coli BL 21 (DE3). The PTD-hEGF fusion protein was induced by 0.3 mmol/L of IPTG expressed in the form of inclusion body with an yield of 40% of the total protein in the cells, and then purified by Ni2+ -NTA affinity chromatography. The SDS-PAGE analysis showed a single fusion protein band with a molecular weight of 16 kD. The amino acid sequence was checked by MALDI-TOF-MS analysis. The genetic engineering PTD-hEGF fusion protein obviously promoted the proliferation and growth of the HEK-293 cells in vitro.
Amino Acid Sequence ; Cell Line ; Cell Proliferation ; drug effects ; Epidermal Growth Factor ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Transduction, Genetic

OBJECTIVETo explore the inhibitory effect of He-Ne laser irradiation on fibroblast growth of hypertrophic scars in culture.

METHODSHe-Ne laser with wavelength of 632.8 nm, power density of 50 mW/cm(2) and doses of 3 J/cm(2), 30 J/cm(2), 90 J/cm(2) and 180 J/cm(2) was used to irradiate human scar fibroblasts in culture 1, 3 and 5 times respectively, and then the cell count and cell cycle analysis were done.

RESULTSRepeated irradiation with He-Ne laser at dose of 180 J/cm(2) three and five times led to an evident decrease in total cell number compared with that of the control group and there was a significant difference (P<0.05). The cell cycle analysis showed after three and five times of irradiation with 180 J/cm(2) He-Ne laser the cell number in S-phase decreased from 51% to 20% and 14% respectively, the cell number in G(0)/G(1) phase increased from 28% to 55% and 60% respectively, and the cell percentage in Sub-G1 phase was 6.7% and 9.8% respectively.

CONCLUSIONSRepeated irradiation with 180 J/cm(2) He-Ne laser can inhibit scar fibroblasts growth in culture. It may be that He-Ne laser irradiation causes cell stagnation in G(0)/G(1) phase and apoptosis.

Cell Division ; radiation effects ; Cells, Cultured ; Cicatrix ; pathology ; Dose-Response Relationship, Radiation ; Female ; Fibroblasts ; cytology ; radiation effects ; Helium ; Humans ; Lasers ; Male ; Neon

OBJECTIVETo reveal the sequence variations of the full coding region of the human alpha (1,beta/1,4) fucosyltransferase 5 gene (FUT5) in a Chinese Han population.

METHODSThe whole coding region of the FUT5 gene was amplified and sequenced in a total of 30 unrelated Chinese Han individuals. The PCR products containing the nucleotide variants observed in the study were subcloned into plasmid pcDNA to determine all potential haplotypes in the investigated population. Genetic polymorphisms of C560T (rs778970) and C484A loci were further analyzed by polymerase chain reaction-restriction fragment length polymorphism(PCR-RLFP) method.

RESULTSIn addition to seven previously reported base substitutions, two novel polymorphisms, namely C484A (Leu162Met) and T684C, were found in the coding region of the FUT5 gene in the 30 individuals. Seven haplotypes were identified by subcloning the variants into plasmid and subsequent DNA sequencing. The allele frequencies in the rs778970 locus in 160 Chinese Han individuals was 0.3031 for 560C and 0.6969 for 560T, while no polymorphism was detected in the C484A locus.

CONCLUSIONThe sequence of the coding region in the human FUT5 gene demonstrated high genetic diversity, and the allelic distribution of the rs778970 locus in the Chinese populations is polymorphic.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; Base Sequence ; Fucosyltransferases ; genetics ; Gene Frequency ; Genetic Variation ; genetics ; Haplotypes ; Humans ; Open Reading Frames ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Population Groups ; genetics ; Sequence Analysis, DNA

Adult ; Asian Continental Ancestry Group ; DNA, Viral ; blood ; Female ; Genes, Viral ; genetics ; Hepatitis B virus ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Protein Isoforms ; genetics ; Viral Core Proteins ; genetics

OBJECTIVESTo analyze the differential expression genes (DEGs) among hepatocellular carcinoma (HCC), para-cancerous tissue (PCT) and normal liver tissue (NLT) and explore the target genes related to the development and progression of HCC.

METHODSThe total RNAs of matched HCC, PCT and NLT of HCC patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis and lab-on-chip. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of HCC and NLT, HCC and PCT were hybridized with Agilent oligo microarray (21,074 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. The selected candidate genes were confirmed by SYBR Green I stained real time RT-PCR.

RESULTS(1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 420 up-regulated genes and 552 down-regulated genes among 2-fold DEGs, including DKK1 (dickkopf homolog 1) which was 5-fold up-regulated; (3) The results of real time RT-PCR, using beta-actin as an internal control, showed that the 2-Delta Ct values of DKK1 in HCC, PCT and NLT were 0.089 504, 0.007,65 and 0.000,631 respectively.

CONCLUSION(1) The high throughput and effective Agilent oligo microarray can screen novel therapy targeted genes by analyzing the DEGs in development and progression of HCC; (2) The development and progression of HCC is a complicated process involving multigenes and multiprocedures; (3) DKK1, as a novel gene, is involved in the development and progression of HCC and may be a new therapy target.

Carcinoma, Hepatocellular ; genetics ; pathology ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; genetics ; pathology ; Oligonucleotide Array Sequence Analysis ; RNA, Neoplasm ; genetics

BACKGROUNDParkinson's disease (PD) is an autosomally inherited neurodegenerative disease in elderly people. The etiology of PD has long been thought to be associated with both genetic and environmental factors. To explore potential genetic risk factors for PD in the northern Han Chinese population, we investigated three single nucleotide polymorphisms (SNPs) (rs4538475, rs11107 and rs12564040) in the BST1, PARK15 and PARK9 genes.

METHODSGenomic DNA from 215 PD patients and 212 matched controls was amplified in two independent PCR systems and subsequently genotyped by digestion with the endonuclease PstI. Genetic parameter and association studies were carried out with SPSS 13.0 and PLINK 1.07 software.

RESULTSWe could accurately detect all genotypes in the three loci with the PCR-RFLP or mismatched PCR-RFLP techniques. The observed heterozygosities of the rs4538475 and rs11107 loci in PD and control groups ranged from 0.460 - 0.481 and 0.410 - 0.441, in BST1, PARK15 respectively, while we detected no heterozygosity at the rs12564040 locus in PARK9. The similar distributions of genotypic frequency between both groups suggest that the three SNPs investigated in this study are unlikely to play roles as common risk factors or pathogenic mutations for PD in northern Han Chinese.

CONCLUSIONThe SNPs investigated in the BST1, PARK15 and PARK9 genes associated with PD susceptibility are not associated with PD in the northern Han Chinese population.

ADP-ribosyl Cyclase ; genetics ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; genetics ; Asian Continental Ancestry Group ; genetics ; F-Box Proteins ; genetics ; Female ; GPI-Linked Proteins ; genetics ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; Parkinsonian Disorders ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; genetics ; Young Adult