Objective To explore the clinical features and surgical management of aged patients with acute cholecystitis,and try to command the opportunity and procedure of laparoseopic cholecystectomy(LC)better.Meth- ods Clinical data of 52 aged cases with acute cholecystits undergone LC were analyzed retrospectively.Results All of the patients(within 48h of the acute attack)were successfully recovered without serious operative complications. Conclusion Aged acute cholecystitis progressed rapidly and its operative difficulty and risk were higher;only if more attention was paid to perioperative managements and operative time and technical skill were mastered,early LC for the patients was safe and feasible.Therefore.it should be recommended in the great majority of cases except the des- perate patients whose general condition was too poor to operate.

Animals ; DNA Replication ; DNA-Directed DNA Polymerase ; genetics ; Expressed Sequence Tags ; Gene Expression ; Humans ; Mutagens ; toxicity ; Mutation ; genetics ; Signal Transduction

OBJECTIVETo optimize the method in the Chinese Pharmacopoeia for determining Salviae Miltiorrhizae Radix et Rhizoma.

METHODTanshinone II(A) and salvianolic acid B were selected as the index in optimization of the sample preparation method of Salviae Miltiorrhizae Radix et Rhizoma in Chinese Pharmacopoeia. Orthogonal test was used to optimize the extraction process of Salviae Miltiorrhizae Radix et Rhizoma, and concentration of contents were detected by high performance liquid chromatography method. A detection of using methanol-water (85: 15) at wavelength of 270 nm was employed for tanshinone II(A) and a detection of using methanol-acetonitrile-formic acid-water (30:10:1: 59) at wavelength of 286 nm was employed for salvianolic acid B.

RESULTThe optimized extraction process of tanshinone II(A) and salvianolic acid B was: extracted by 90% methanol and reflux twice (0.5 h each time) at 75 degrees C, extracted by 70% methanol and reflux twice (1.5 h each time) at 75 degrees C, respectively.

CONCLUSIONOptimized extraction and determination methods could be used to reflect the content of tanshinone II(A) and salvianolic acid B in Salviae Miltiorrhizae Radix et Rhizoma more accurately and efficiently.

Benzofurans ; analysis ; Chromatography, High Pressure Liquid ; Diterpenes, Abietane ; analysis ; Rhizome ; chemistry ; Salvia miltiorrhiza ; chemistry ; Temperature

OBJECTIVETo investigate the function of POLH(polymerase eta) through establishment of the POLH gene-blocked cell line FL-POLH(-).

METHODSA mammalian expression vector expressing antisense POLH gene fragment pMAMneo-amp-POLHA (-) was constructed by cloning the 1473 - 2131 fragment of POLH gene into the mammalian expression vector pMAMneo-amp(-) in antisense orientation. The FL cells were transfected with this antisense RNA expressing vector and selected by G418. The mutation assay was conducted using the shuttle plasmid pZ189.

RESULTThe spontaneous mutation frequency of SupF tRNA gene in the plasmid replicated in the FL-POLH(-) was 13.5 x 10(-4), while it was 4.9x10(-4) and 3.7x10(-4) in the control cells FL and FL-M, respectively. The nontargeted mutation frequency of SupF tRNA gene decreased in the plasmid replicated in these cell lines pretreated with MNNG.

CONCLUSIONPOLH plays an important role in maintenance of genetic stability and genesis of nontargeted mutation.

Antisense Elements (Genetics) ; pharmacology ; Cell Line ; DNA-Directed DNA Polymerase ; genetics ; physiology ; Methylnitronitrosoguanidine ; toxicity ; Mutagenesis

OBJECTIVETo understand the up regulatory mechanism of human REV3 gene induced by the chemical carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG).

METHODSBioinformatic analysis of human REV3 gene promoter region was based on BLAST alignment, promoter prediction software and recognition of transcriptional factor binding sites. Cloning of human REV3 gene promoter region was performed by nested PCR. Response of human REV3 gene promoter to the chemical carcinogen MNNG was measured by transient transfection assay based on the dual luciferase reporter assay system.

RESULTBioinformatic analysis showed that human REV3 gene promoter region was located on chromosome 6 PAC clone RP3-415N12, and that the hypothetical promoter region contained promoter sequences, rich CpG islands, and putative recognition sites for several transcriptional factors, including AP-1/c-Jun/c-Fos, AP-2, STAT, CREBP, and NF-kappaB. Reconstructed reporter plasmid pGL3- 2582 was established by inserting 2582 nucleotides from the promoter region into the luciferase reporter vector pGL3-Basic. Transient transfection assay showed the hypothetical REV3 promoter region had promoter function, and it responded to MNNG treatment (P<0.01).

CONCLUSIONHuman mutator REV3 gene promoter region has been successfully cloned. The response of REV3 promoter region to MNNG suggests that REV3 gene can be regulated at transcriptional level under conditions of genotoxic stress.

Base Sequence ; Binding Sites ; Cloning, Molecular ; Computational Biology ; DNA-Directed DNA Polymerase ; genetics ; Humans ; Methylnitronitrosoguanidine ; toxicity ; Molecular Sequence Data ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Saccharomyces cerevisiae Proteins ; genetics

OBJECTIVETo investigate the effect of MNNG on some of the transcription factors such as NF- kappaB, CREB, AP-1 and c-Myc.

METHODSThe activities of these transcription factors were measured by transient transfection assay of SEAP vectors.

RESULTThe expressions of AP-1, CREB and NF- kappaB driven reporter genes were elevated for about 1.3, 1.4 and 1.3 times in MNNG-treated cells, respectively, as compared to untreated controls. However, the exposure of MNNG had no effect on the activity of c-Myc.

CONCLUSIONThe activation of certain transcription factors might be involved in the process of untargeted mutation induced by low concentration of MNNG treatment.

Animals ; Cercopithecus aethiops ; Cyclic AMP Response Element-Binding Protein ; drug effects ; Methylnitronitrosoguanidine ; toxicity ; Mutation ; NF-kappa B ; drug effects ; Proto-Oncogene Proteins c-myc ; drug effects ; Transcription Factor AP-1 ; drug effects ; Transcription Factors ; drug effects ; Vero Cells

OBJECTIVETo understand benzo[a]pyrene (B[a]P) mediated cellular responses, and to provide clues to explore molecular mechanism of mutagenesis and carcinogenesis induced by B[a]P.

METHODSTwo-dimensional electrophoresis (2-DE) was used to investigate the protein expression levels of FL cells after B[a]P exposure, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) combined with database search was applied to identify the differentially expressed proteins.

RESULTStatistical analysis showed that the volumes of 47 protein spots were altered after B[a]P treatment (P<0.05) and 23 proteins were successfully identified, including zinc finger proteins, SWI/SNF related protein, Bromo domain containing domain and other proteins.

CONCLUSIONThese affected proteins may be involved in the cellular responses to B[a]P exposure, and may mediate the B[a]P induced mutagenesis and carcinogenesis.

Amnion ; chemistry ; cytology ; drug effects ; Benzo(a)pyrene ; toxicity ; Cells, Cultured ; DNA Repair ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Proteomics ; Zinc Fingers

OBJECTIVETo investigate the protein profile after treatment of low concentration of N- methyl-N'-nitro-N-nitrosoguanidine (MNNG) in human FL cells.

METHODSAfter MNNG treatment, whole cellular proteins were separated using two-dimensional gel electrophoresis and visualized by silver staining; the digitized images were analyzed with 2D analysis software. The differentially expressed protein spots were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).

RESULTMore than 60 proteins showed significant changes in MNNG-treated cells compared to control cells (DMSO treatment). There were 18 protein spots detected only after MNNG treatment, while 13 protein spots were detected only in the control cells. Moreover, the levels of another 31 proteins were either increased or decreased in MNNG-treated FL cells. And some of the proteins were identified by MALDI-TOF-MS.

CONCLUSIONThere are significant alterations of protein profile after MNNG attack.

Amnion ; chemistry ; cytology ; drug effects ; Cells, Cultured ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Methylnitronitrosoguanidine ; toxicity ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Adolescent ; Brain Neoplasms ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Melanoma ; metabolism ; pathology ; Melanoma-Specific Antigens ; metabolism ; Neurilemmoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Vimentin ; metabolism

Animals ; Apoptosis ; physiology ; Cellular Senescence ; physiology ; Humans ; Neoplasms ; pathology ; Sphingolipids ; chemistry ; metabolism ; physiology