OBJECTIVETo investigate the effects of matrine on the cell cycle and apoptosis in human colon adenocarcinoma SW620 cells and explore the possible mechanisms.

METHODSThe effect of matrine on cell proliferation was assessed using MTT assay, and the cell cycle arrest induced by matrine was determined by flow cytometry. The changes of cell morphology were observed through optical microscope, fluorescence microscope and electron microscope, and the cell apoptosis was detected using Annexin V-FITC apoptosis assay.

RESULTSMatrine inhibited the proliferation of SW620 cells in a dose- and time-dependent manner. Compared with the control group, the matrine-treated cells showed increased cell percentage arrested in G 0/G1 phase with decreased S-phase cells. Morphologically, the SW620 cells treated with matrine exhibited cell shrinkage, cell size reduction, plasma condensation, cytoplasmic vacuolar changes, and formation of apoptotic body with also the presence of the signet-ring cells, all typical of apoptotic cells.

CONCLUSIONMatrine exposure of SW620 cells inhibits the cell proliferation, causes cell cycle arrest at G 0/G1 phase, and induces apoptosis in a dose- and time- dependent manner.

Adenocarcinoma ; pathology ; ultrastructure ; Alkaloids ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Shape ; drug effects ; Colorectal Neoplasms ; pathology ; ultrastructure ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Quinolizines ; pharmacology

OBJECTIVEExpression of vimentin in carcinoma cells is a marker for poor prognosis in patients. The aim of this investigation was to assess the influence of vimentin on the characteristics of carcinoma cells.

METHODSThe full-length vimentin gene open reading frame (1401 base pairs) was cloned into the plasmid vector pcDNA 3.1 (+), and these vectors were used to stably transfect the human hepatocellular carcinoma HepG2 cell line. Vimentin gene expression was evaluated by RT-PCR and Western blot. Proliferative activity and invasive potential of tumor cells were determined by the CellTiter 96 aqueous one solution cell proliferation assay and BioCoat GFR Matrigel invasion chamber, respectively.

RESULTSDNA sequencing and restriction endonuclease digestion analysis demonstrated that the recombinant vector was correctly cloned. The stable cell line demonstrated a higher vimentin RNA and protein expression. However, both proliferative and invasive abilities of the cells were reduced in vitro ( P < 0.05).

CONCLUSIONA recombinant plasmid pcDNA3. 1-VIM is successfully constructed and a carcinoma cell line HepG2-pV highly expressing vimentin is obtained. Recombinant vimentin protein suppresses the proliferative and invasive abilities of HepG2 cells, suggesting that it might decrease malignant phenotype of tumor cells in vitro. This work makes a foundation for further study on the relationship between vimentin and biological phenotype of carcinoma cells.

Cell Movement ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Hep G2 Cells ; Humans ; Neoplasm Invasiveness ; Open Reading Frames ; genetics ; Plasmids ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Vimentin ; genetics ; metabolism

OBJECTIVETo search for the genes which could interact with newly found homo sapiens cross-immune reaction antigen (PCIA1) gene and accordingly to provide insights into the study of the gene function.

METHODSThe Stratagene's BacterioMatch Two-Hybrid System and BacterioMatch Fetal Kidney Library were adopted and the recombinant bait plasmid pBT-PCIA1 was cotransformated with the target plasmid pTRG-cDNA library DNA into the reporter stain. After screening and isolation of positive pTRG clones, the target genes were identified by DNA sequencing and bioinformation analysis.

RESULTSAmong all the seven detected target genes, three genes' function were not known, the other four genes had important functions. Their mutations or abberant expression resulted in severe diseases and overexpression of ACTN4 (actinin, alpha 4), PSAP (prosaposin) or EIF3S10 (eukaryotic translation initiation factor 3, subunit 10 theta) could promote tumor development and progression.

CONCLUSIONThe bacterial two-hybrid system technique is an efficient method, which can provides insights into the study of novel genes' function by detecting protein-protein interactions. This study indicates that PCIA1 gene expression correlates with tumor formation, invasion and metastasis.

Bacteria ; genetics ; metabolism ; Computational Biology ; DNA Restriction Enzymes ; metabolism ; Gene Library ; Genetic Vectors ; Humans ; Neoplasms ; genetics ; pathology ; Plasmids ; genetics ; Sequence Analysis, DNA ; Two-Hybrid System Techniques

OBJECTIVETo investigate whether the variants A(-6)G and A(-20)C of angiotensinogen (AGT) gene are involved in the pathogenesis of essential hypertension in Kazakans.

METHODST his case control study recruited 125 subjects with hypertension and 74 normotensive subjects from Kazakans of Xinjiang. Genomic DNA from leukocytes was analyzed for genetic variants A(-6)G and A(-20)C in 5' upstream core promoter of AGT gene by polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP), restriction fragment length polymorphism (RFLP) and automatic sequencing.

RESULTS(1)There were only A(-6)G and A(-20)C variants in the -164 to +73 region of Kazakans' AGT gene. (2) The distributions of genotypes AA, AG, GG at locus -6 of AGT gene showed significant difference between the hypertensive group (0.39, 0.45, 0.16) and the normotensive group (0.49, 0.49, 0.02; Chi2=8.56, P=0.014). There were evident differences in the frequencies of the -6A and the -6G allele of the two groups (0.62, 0.38 and 0.73, 0.27; Chi2=5.35, P=0.021). (3) No significant difference was observed in the distribution of genotypes AA, AC, CC at locus -20 of AGT gene between the hypertensive group (0.69, 0.26, 0.05) and the normotensive group (0.65, 0.32, 0.03; Chi2=2.42, P=0.30). There was no distinct difference in the frequencies of the -20A allele and the -20C allele of the two groups (0.82, 0.18 and 0.82, 0.18; Chi2=0, P=0.99). (4) No significant difference was found at the levels of systolic and diastolic blood pressure between the groups corresponding to genotypes at the loci -6 and -20 of AGT gene.

CONCLUSIONThe results suggest that the polymorphism of A(-6)G in 5' upstream core promoter of the AGT gene may be involved in the pathogenesis of essential hypertension in Kazakans, while the A(-20)C variant may not play an important role in the etiology of essential hypertension in Kazakans.

5' Flanking Region ; genetics ; Adult ; Alleles ; Angiotensinogen ; genetics ; Base Sequence ; Blood Pressure ; physiology ; Case-Control Studies ; China ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; genetics ; physiopathology ; Male ; Middle Aged ; Molecular Sequence Data ; Polymorphism, Genetic ; Polymorphism, Single-Stranded Conformational ; Promoter Regions, Genetic ; genetics

The aim of this study was to investigated the biological diversity, antibacterial activites and the plant growth-promoting traits of endophytic fungi of sandal (Santalum album), and to assess their potential in the development of antibacterial substances and rapid cultivation of sandal. The results of isolation and taxa analysis of endophytic fungi from sandal showed that 325 strains of endophytic fungi belonging to 16 genera of endophytic fungi were isolated from sandal (of which 86 from roots, 105 from stems and 134 from leaves). The isolation rate and colonization rate of endophytic fungi in different sandal parts showed the same pattern of change: leave>stems>roots. The diversity index of endophytic fungi in sandal roots was significantly higher than that of stems and leaves. The dominant endophytic fungi of sandal roots, stems and leaves showed significant differences. The dominant endophytic fungi of roots were Fusarium (50.00%) and Alternaria (10.47%), Alternaria (58.11%) and Acremonium (20.00%) for stems, and Pantoea (74.63%) for leaves. The antibacterial activity of 40 representative strains of sandal endophytic fungi were analyzed and the results showed that 90% of endophytic fungi exhibited inhibitory activity against at least one of the tested bacteria strains, and the strains with inhibitory activity to Escherichia coli, Enterobacter aerogenes, Shigella dysenteriae, Salmonella typhimurium, Staphylococcus aureus, and Bacillus subtilis accounted for 45.0%, 30%, 47.5%, 55%, 72.5%, and 62.5%, respectively. The sandal fungal endophytes with plant growth-promoting characteristics were screened, and 5 strains of endophytic fungi with phosphorus-solubilizing activity, 8 strains of endophytic fungi producing IAA, and 4 strains of endophytic fungi producing siderophores were found. Among them, endophytic fungus Monilia sp TXRF45 clould produced IAA and siderophores, and also show phosphate-solubilizing activity. The results indicated that the endophytic fungi of Sandal were rich in species diversity and their distribution had a certain tissue specificity. Some strains showed good antibacterial activity and growth-promoting properties, which could potentially applicable for the development of antibacterial substances and rapid cultivation of sandal.

The aim of this study was to investigate the effect of WR2721(amifostine) against bone marrow hematopoietic damage of mice exposed to 6.5 Gy of (60)Co-γ ray. A total of 60 C57/BL6J mice was divided into 3 groups:normal group (mice were injected with physiological salt solution), irradiation group (mice were injected with physiologic salt solution before irradiation) and WR2721 group (mice were injected with WR2721 before irradiation). The WBC, neutrophil (Neut), Plt and RBC levels in peripheral blood of 3 group mice were counted within 60 days after irradiation; the bone marrow nuclear cells (BMNC) were counted at 2 and 24 hours after irradiation; the hematopoietic stem/progenitor cell (LK/LSK) level and colony formation capability were detected by flow cytometry at 2 and 24 hours after irradiation. The results indicated that the counts of WBC and neut at 4 and 18 days, Plt at 7-18 days and RBC at 10-30 day after irradiation in WR2721 group were higher than those in irradiation group (P < 0.05); the BMNC, LSK and LK levels obviously increased at 24 hours after irradiation (P < 0.05), the CFU-GEMM, CFU-GM, CFU-MK BFU-E and CFU-E all significantly increased at 2 and 24 hours after irradiation (P < 0.01), as compared with irradiation group. It is concluded that WR2721 can effectively alleviate early hematopoietic damage and promote the fast recovery of peripheral blood cells in mice exposed to γ-ray, suggesting that the WR2721 has significant radioprotective effect on hematopoietic system.
Amifostine ; pharmacology ; Animals ; Blood Cell Count ; Bone Marrow Cells ; cytology ; drug effects ; radiation effects ; Gamma Rays ; Hematopoietic Stem Cells ; cytology ; drug effects ; radiation effects ; Male ; Mice ; Mice, Inbred C57BL ; Radiation-Protective Agents ; pharmacology ; Whole-Body Irradiation

Many phytochemicals show promise in cancer prevention and treatment, but their low aqueous solubility, poor stability, unfavorable bioavailability, and low target specificity make administering them at therapeutic doses unrealistic. This is particularly true for (-)-epigallocatechin gallate, curcumin, quercetin, resveratrol, and genistein. There is an increasing interest in developing novel delivery strategies for these natural products. Liposomes, micelles, nanoemulsions, solid lipid nanoparticles, nanostructured lipid carriers and poly (lactide-co-glycolide) nanoparticles are biocompatible and biodegradable nanoparticles. Those nanoparticles can increase the stability and solubility of phytochemicals, exhibit a sustained release property, enhance their absorption and bioavailability, protect them from premature enzymatic degradation or metabolism, prolong their circulation time, improve their target specificity to cancer cells or tumors via passive or targeted delivery, lower toxicity or side-effects to normal cells or tissues through preventing them from prematurely interacting with the biological environment, and enhance anti-cancer activities. Nanotechnology opens a door for developing phytochemical-loaded nanoparticles for prevention and treatment of cancer.
Antineoplastic Agents, Phytogenic ; administration & dosage ; therapeutic use ; Drug Carriers ; Humans ; Materials Testing ; Nanoparticles ; Neoplasms ; drug therapy ; Phytochemicals ; administration & dosage ; therapeutic use ; Plant Extracts ; administration & dosage ; therapeutic use

Objective:To investigate the taxonomic structure and diversity of endophytic fungi from Datura metel and screen the strains with anti-dermatophyte activities， so as to provide resources for the development of new lead compounds against dermatomycosis. Method:Endophytic fungi were isolated from the roots， stems and leaves of D. metel after tissue block incubation and then identified by morphological analysis and rDNA-internal transcribed spacer（ITS） sequencing. Their anti-dermatophyte activities were detected by agar diffusion assay. Result:A total of 292 strains of endophytic fungi were isolated from D. metel， belonging to 34 genera， with Fusarium （72.97%） in roots， Fusarium （37.25%） and Plectosphaerella （28.43%） in stems， and Colletotrichum （39.66%） in leaves as the dominant species. The isolation rate （89.23%）， colonization rate （84.62%）， and diversity index （1.82） of endophytic fungi in leaves were significantly higher than those in roots （70.48%， 70.48% and 1.23） and stems （69.39%，68.03% and 1.64）. The determination of anti-dermatophyte activities of 35 endophytic fungal fermented filtrates showed that the strains exhibiting inhibitory activities against Microsporum canis， Trichosporon mucoides， Trichophyton rubrum and Candida albicans accounted for 97.14%， 71.43%， 45.71%， and 25.71%， respectively. Among them， six strains （17.14%）， namely Fusarium sp. R1， Penicillium sp. R5， Aspergillus sp. R7， Metarhizium sp. S18， Diaporthe sp. S19， and Glomerella sp. L57， all inhibited the four types of cutaneous fungal pathogens. Conclusion:The endophytic fungi in D. metel are diverse， and the proportion of endophytic fungi possessing anti-dermatophyte activities is high， allowing them to serve as potential resources for the development of new anti-dermatophyte agents.