Adrenal Cortex Hormones ; therapeutic use ; Anemia, Hemolytic, Autoimmune ; diagnosis ; drug therapy ; Female ; Humans ; Infant ; Treatment Outcome

The paper is about the systematic studies of biological characteristics of 15 stains rhizobia isolated purified from Acacia confusa grew in Guangxi karst environment.The results showed that there were typical characteristics of rhizobia.there were negative reaction about use of 3-ketolactose and beef extract peptone nutrient agar medium,and positive reaction about use of starch and citrate medium,and produce acid in reaction of BTB and litmus milk medium,(NH4)2HPO4 was used as nitrogen sources and both four monosaccharides and three disaccharides could be utilized as carbon sources in 15 strains rhizobia isolated Acacia confuse.Among the 15 strains for the tests,11 strains could deoxidize the nitrate of medium into nitrite,14 strains could grow well on NaCl solution concentration 3.0 %～4.0 %,14 strains could grow at 39℃,13 strains may grow on highest pH12 and 4 strains on lowest pH4 cultrue medium.15 strains can grow in 10% and 11 strains in 10%～30% of CaCO3 solution concentration.

Background Histone deacetylase inhibitors can regulate gene expression through modulation of the degree of acetylation of histone and non-histone,thus affecting cell proliferation,survival and chemosensitivity.Histone deacetylase inhibitors combined with paclitaxel may enhance the inhibitory effect of drugs on lung cancer cells.This study aimed to observe the effect of trichostatin A (TSA)/paclitaxel on the proliferation and apoptosis in human A549 lung adenocarcinoma cells,and to investigate its mechanism.Methods A549 cells were cultured in Dulbecco modified Eagle's medium (DMEM) in the presence of paclitaxel and the histone deacetylase inhibitor TSA,and the growth curve was obtained by trypan-blue exclusion assay and cell count.Apoptosis was assessed using Hoechst 33258 staining and flow cytometry analysis,and cell cycle was detected by flow cytometry analysis.The proteins poly ADP-ribose polymerase (PARP),caspase-3,survivin,and tubulin acetylation were detected by Western blotting.Results A significant reduction of proliferation was observed in A549 lung adenocarcinoma cells treated by paclitaxel or TSA.Combined treatment with TSA/paclitaxel caused the greatest inhibition of cell proliferation.The combined treatment with TSA and paclitaxel induced more severe apoptosis,and significantly more cells were arrested in Gz/M phase (P ＜0.05) then with a single drug.Using Western blotting,we demonstrated that treatment with TSA/paclitaxel led to synergistic increase in acetylated tubulin,PARP,caspase-3,and reduced the expression of survivin.Conclusion TSA and paclitaxel have a synergistic activity that can inhibit cell growth and induce apoptosis.

BACKGROUNDHistone deacetylase inhibitors can regulate gene expression through modulation of the degree of acetylation of histone and non-histone, thus affecting cell proliferation, survival and chemosensitivity. Histone deacetylase inhibitors combined with paclitaxel may enhance the inhibitory effect of drugs on lung cancer cells. This study aimed to observe the effect of trichostatin A (TSA)/paclitaxel on the proliferation and apoptosis in human A549 lung adenocarcinoma cells, and to investigate its mechanism.

METHODSA549 cells were cultured in Dulbecco modified Eagle's medium (DMEM) in the presence of paclitaxel and the histone deacetylase inhibitor TSA, and the growth curve was obtained by trypan-blue exclusion assay and cell count. Apoptosis was assessed using Hoechst 33258 staining and flow cytometry analysis, and cell cycle was detected by flow cytometry analysis. The proteins poly ADP-ribose polymerase (PARP), caspase-3, survivin, and tubulin acetylation were detected by Western blotting.

RESULTSA significant reduction of proliferation was observed in A549 lung adenocarcinoma cells treated by paclitaxel or TSA. Combined treatment with TSA/paclitaxel caused the greatest inhibition of cell proliferation. The combined treatment with TSA and paclitaxel induced more severe apoptosis, and significantly more cells were arrested in G2/M phase (P < 0.05) then with a single drug. Using Western blotting, we demonstrated that treatment with TSA/paclitaxel led to synergistic increase in acetylated tubulin, PARP, caspase-3, and reduced the expression of survivin.

CONCLUSIONTSA and paclitaxel have a synergistic activity that can inhibit cell growth and induce apoptosis.

Acetylation ; Adenocarcinoma ; drug therapy ; pathology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Histone Deacetylase Inhibitors ; pharmacology ; Humans ; Hydroxamic Acids ; pharmacology ; Lung Neoplasms ; drug therapy ; pathology ; Paclitaxel ; pharmacology ; Tubulin ; metabolism

To construct Fv antibodies against H5N1 Avian influenza virus hemagglutinin,extracted mRNA from B lymphoblastoid cell lines secreting anti-HA antibodies was used and the VH and VL genes were amplified by RT-PCR and linked together by splicing overlap extension (SOE) with (Gly4 Ser)3 linker. The recombinant plasmid was then transformed to E. coli BL21(DE3) and sequence analysis indicated the total length of scFv was 714 bp and the expression of Fv was validated by PAGE and Western blot.
Animals ; Antibodies ; genetics ; metabolism ; pharmacology ; Birds ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation ; Hemagglutinins ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; metabolism ; Influenza A Virus, H5N1 Subtype ; drug effects ; immunology ; Influenza in Birds ; virology ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; immunology

OBJECTIVETo investigate the effect of trichostatin A (TSA)/paclitaxel on the growth and apoptosis in human lung adenocarcinoma cell line A549 cells.

METHODSHuman lung adenocarcinoma A549 cells were cultured in DMEM in the presence of paclitaxel and the histone deacetylase inhibitor trichostatin A, and the growth curve was obtained by trypan-blue exclusion assay and cell count. Apoptosis was assessed using Hoechst 33258 staining and flow cytometry, and cell cycle was detected by flow cytometry analysis. The proteins of PARP, caspase-3, survivin and tubulin acetylation were detected by Western blotting.

RESULTSSignificant growth reduction was observed in the A549 cells following treatment with paclitaxel or the histone deacetylase inhibitor TSA. The combined treatment with TSA/paclitaxel caused the highest inhibition of cell growth. The apoptosis rate of A549 cells treated with TSA or paclitaxel for 24 hours was (17.6 ± 1.8)% and (39.2 ± 3.7)%, respectively, but a significantly higher apoptosis rate was (64.2 ± 4.2)% was induced by combined treatment with TSA and paclitaxel. In contrast with the control group, the cell cycle was markedly arrested at G2/M phase in the TSA and paclitaxel group (P < 0.05). The Western blot analysis demonstrated that treatment with TSA/paclitaxel led to a synergistic increase of acetylated tubulin, PARP and caspase-3, and reduced the expression of survivin.

CONCLUSIONTSA or paclitaxel alone can inhibit the cell growth and induce apoptosis, and the combination of TSA and paclitaxel exerts a synergistic effect on the growth and apoptosis in lung adenocarcinoma cells.

Acetylation ; Adenocarcinoma ; metabolism ; pathology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Synergism ; Histone Deacetylase Inhibitors ; pharmacology ; Humans ; Hydroxamic Acids ; pharmacology ; Inhibitor of Apoptosis Proteins ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Paclitaxel ; pharmacology ; Poly(ADP-ribose) Polymerases ; metabolism ; Tubulin ; metabolism ; Tubulin Modulators ; pharmacology

AIM To study the chemical constituents from the fruits of Psidum littorale Raddi.METHODS The ethyl acetate and methanol fractions of 90％ ethanol extract from P.littorale were isolated and purified by silica,Sephadex LH-20,D-101 macroporous resin and preparative HPLC column,then the structures of obtained compounds were identified by spectral data.RESULTS Twelve compounds were isolated and identified as lupeol (1),betulin (2),nigaichigoside F1 (3),2oα-hydroxybetulinic acid (4),(-)-episyringaresinol (5),(+)-syringaresinol (6),(+)-isolariciresinol-9'-O-β-glucopyranoside (7),(7S,8R)-urolignoside (8),benzyl-β-D-glucopyranoside (9),butanedioic acid (10),β-sitosterol (11),daucosterol (12).CONCLUSION Compounds 1-9 are isolated from genus Psidium for the first time.

AIM To study the phenylethanoid glycosides from Verbenae Herba.METHODS The 80%ethanol extract from Verbenae Herba was isolated and purified by silica gel,Sephadex LH-20,TLC and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as verbofficoside A(1),cistanoside D(2),epimeredinoside A(3),verbascoside(4),isoverbascoside(5),cistanoside C(6),cistanoside F(7),decaffeoylacteoside(8),jionoside C(9).CONCLUSION Compound 1 is a new compound.Compounds 3 and 6-9 are isolated from this plant for the first time.

OBJECTIVETo observe the effects of ganoderma lucidum spores (GLS) on mitochondrial calcium ion and cytochrome C in the epididymal cells of type 2 diabetes rats.

METHODSFifty adolescent rats were randomly divided into a model group (n=20), a GLS group (n=20) and a control group (n=10). The animals of the former two groups were injected with 2% STZ via vena caudalis for one time to induce type 2 diabetes. Then the model group was given high-fat-sugar diet, the GLS group high-fat-sugar diet + GLS (250 mg/kg x d), and the control group normal diet + CA-citrate sodium buffer. The bilateral epididymides were obtained 10 weeks later and the contents of mitochondrial calcium and cytochrome C detected.

RESULTSType 2 diabetes models were successfully constructed. The content of mitochondrial calcium in the epididymal cells was significantly higher in the model group ([3.279 +/- 0.502] mg/L) than in the control group ([2.606 +/- 0.048] mg/L, P < 0.01), with no significant difference between the GLS group ([2.693 +/- 0. 196] mg/L) and the control (P > 0.05). In the model group, the content of mitochondrial cytochrome C ([3.213 +/- 1.511] micromol/L) was significantly lower (P < 0.05) while that of cytoplasm cytochrome C ([2.484 +/- 0.661] micromol/L) significantly higher (P < 0.05) than in the control ([5.688 +/- 1.679] micromol/L and [1.574 +/- 0.329] micromol/L, respectively). In the GLS group, the content of mitochondrial cytochrome C ([5.258 +/- 1.560] micromol/L) was higher, with no significant difference (P > 0.05), and that of cytoplasm cytochrome C ([1.727 +/- 0.396] micromol/L) significantly lower than in the model group (P < 0.05), but the difference between the GLS and the control group was not significant (P > 0.05).

CONCLUSIONWith disequilibrium of calcium homeostasis and damage to mitochondria, there might be excessive apoptosis in the epididymal cells of type 2 diabetes rats. Ganoderma lucidum spores could protect epididymal cells and counteract their apoptosis in diabetic condition.

Animals ; Calcium ; metabolism ; Cytochromes c ; metabolism ; Diabetes Mellitus, Experimental ; therapy ; Diabetes Mellitus, Type 2 ; therapy ; Epididymis ; cytology ; pathology ; Male ; Mitochondria ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Reishi ; physiology ; Spores, Fungal

OBJECTIVETo study the rabies molecular biology features in animals between high incidence area of rabies and no rabies cases area in Hunan.

METHODSdetect saliva of dogs and brains of dogs and cats by direct immunofluorescence assay, review positive samples by RT-PCR, sequencing extract RNA virus for genetic analysis.

RESULTS12 were detected rabies virus antigen and positive nucleoside acid in 82 dogs from Wugang city also 1 in 17 from Dongkou county; the positive rate: Wugang 14.63 percent, Dongkou 5.88 percent. No rabies virus was detected in 67 samples of dog brains from Fenghuang County. Also none in 28 samples of cat brains. Amplificating N gene of rabies virus from positive samples of dog brain's tissue (No Wg13, Dk13) by RT-PCR, it shows that homology of nucleoside acid between two strain of virus is 99.4 percent; also 99.1 percent of amino acid. The homology of nucleoside acid (amonio acid) among Wg13 stain and Chinese strain CTN and aG strain is 89.4 percent (98.2 percent) and 86.1 percent (95.1 percent); The homology of nucleoside acid (amonio acid) among Dk13 stain Chinese strain CTN and aG strain is 89.1 percent (98.0 percent), 86.1 percent (94.9 percent).Compare with isolated rabies virus from abroad, the homology between two strains and Indonesia is 92.8 percent and 93.2 percent, the most similar of them. The strains isolated from other countries including Japan, Sri Lanka and India are relatively lower; The sequence of gene Wg13 and Dk13 were taken replacement of amino acid.

CONCLUSIONTwo strains are belong to type I rabies virus, comparing its N gene with current using vaccine strains, both are in same group, and homology are relatively higher.

Animals ; Cats ; Dogs ; Genes, Viral ; Phylogeny ; Rabies virus ; classification ; genetics ; Reverse Transcriptase Polymerase Chain Reaction