Teaching principles and class content were stated for a experimental course of common pathogenic bacteria detection methods for the undergraduate student in food quality and safety major. They include course material preview, advanced teaching methods, combination of teaching and research, graduate teaching assistant, experimental reports writing and experimental skills evaluation. All these means lead to a good teaching outcome.

OBJECTIVETo study the mechanism of microelement including Cu and Zn on the accumulation of three danshinones in Salviae miltiorrhizae root and build a theory basis for its good quality and high yield.

METHODSand culture experiments were conducted to study the effect of Cu and Zn on the accumulation of three danshinones and oxidase including peroxidase and polyphenol oxidase activity in the plant root. The correlation between available Cu and Zn contents in matrix and oxidase activity in the plant root and, the correlation between available Cu and Zn contents in matrix and contents of tanshinones in the root were discussed.

RESULTContents of danshinones in the root increased with the increasing of Cu and Zn concentration. Dynamic monitoring on contents of dan-shinones of the plant roots growing in the pots with different Cu and Zn concentration in the whole growing season showed that the contents of danshinones for 60 days were the lowest, for 120 days the highest and then dropped for 150 days. In addition, among available Cu and Zn contents of matrix, oxidase including peroxidase and polyphenol oxidase activity and contents of tanshinones in the root,the correlation between two factors were significant difference (P < 0.05 or P < 0.01).

CONCLUSIONThe mechanism of Cu and Zn on the accumulation of danshinones may be that Cu and Zn improve the activity of peroxidase and polyphenol oxidase, which promote transformation of phenolic compounds to terpenes and therefore to increase contents of danshinones.

Catechol Oxidase ; metabolism ; Copper ; metabolism ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; analysis ; metabolism ; Peroxidase ; metabolism ; Phenanthrenes ; metabolism ; Plant Proteins ; metabolism ; Salvia miltiorrhiza ; enzymology ; metabolism ; Zinc ; metabolism

OBJECTIVETo explore the effect of perioperative nutrition support on nutritional condition and complications of the patients with postoperative pancreatic head cancer.

METHODSThirty four patients received perioperative nutrition support, including enteral nutrition and parenteral nutrition (treatment group). Forty eight patients received routine postoperative parenteral nutrition (control group). According to the operative method, these two groups were further divided into two sub-groups: (1) pancreaticoduodenectomy (PD) subgroup, including 13 cases from treatment group, and 24 cases from control group; (2) palliative operation subgroup, including 21 cases from treatment group, and 24 cases from control group. Body weight, total protein (TP), serum albumin (ALB), and the complications after operation were compared.

RESULTSThe concentrations of ALB and TP in the treatment group were significantly higher than those in the control group (P< 0.05). Body weight and TP of the patients received PD in the treatment group were significantly better than those of the control group (P < 0.05).

CONCLUSIONPerioperative nutrition support can improve postoperative nutritional condition and reduce the postoperative complications in patients with pancreatic head cancer.

Adult ; Aged ; Combined Modality Therapy ; Enteral Nutrition ; Female ; Humans ; Male ; Middle Aged ; Nutritional Support ; methods ; Pancreatic Neoplasms ; surgery ; therapy ; Pancreaticoduodenectomy ; Parenteral Nutrition ; Postoperative Complications ; prevention & control

OBJECTIVETo investigate the protective effects of oxymatrine on chronic heart failure induced by isoproterenol (ISO) and to observe its effects on ADMA metabolism pathway in ISO-induced chronic heart failure in rats.

METHODMale Sprague-Dawley rats were given oxymatrine (100,50 mg kg-1) orally for 14 days. Heart failure was induced in rats by subcutaneous injection of isoproterenol (5 mg kg-1 d-1 ) at the 8th day for 1 week. Serum parameters, haemodynamic parameters, Heart weight, and histopathological variables were analysed. Expression of protein levels were measured by Western blot.

RESULTOxymatrine (100,50 mg kg-1) significantly attenuated serum content of cTn I, improved left ventricle systolic and diastolic function and left ventricular remodeling, reduced the ISO-induced myocardial pathological changes compared with ISO group. In addition, oxymatrine (100,50 mg kg-1) significantly reduced serum level of ADMA (P <0. 01), normalize the reduced dimethylarginine dimethylaminohydrolase 2 (DDAH2) expression (P <0. 01) , but had no effect on the isoproterenol-induced upregulated protein arginine methyltransferases 1 expression.

CONCLUSIONOxymatrine could ameliorate the experimental ventricular remodeling in ISO-induced chronic heart failure in rats and the mechanism involved in reducing serum content of ADMA and increased DDAH2 expression.

Alkaloids ; pharmacology ; therapeutic use ; Amidohydrolases ; metabolism ; Animals ; Arginine ; analogs & derivatives ; blood ; metabolism ; Chronic Disease ; Gene Expression Regulation, Enzymologic ; drug effects ; Heart Failure ; drug therapy ; metabolism ; pathology ; physiopathology ; Hemodynamics ; drug effects ; Isoproterenol ; adverse effects ; Male ; Organ Size ; drug effects ; Quinolizines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Troponin I ; metabolism

The purpose of this study was to establish a SYBR Green I real-time quantitative RT-PCR method for investigating the correlation between CML28 mRNA expression levels and relapse of leukemia after allo-hematopoietic stem cell transplantation (HSCT). pcDNA3.1HisA-CML28 plasmid had been constructed as the standard template. Serial monitoring of CML28 mRNA levels by SYBR Green I real-time quantitative RT-PCR technique was performed in 14 patients, including 10 patients with CML and 3 patients with AML, 1 patient with Ph(+) ALL. The results showed that the sensitivity of the established method was at 10(-4) (0.05 ng) level, with interassay variation and intraassay variation of standard samples both below 10%. The CML28 was highly expressed in AML and CML-BP or AP. CML28 level in newly diagnosed group was (6.58 +/- 2.34) x 10(-2), in pre-conditioning regimen group was (2.19 +/- 0.32) x 10(-2), in group that 1 month after HSCT was (1.35 +/- 1.28) x 10(-2), in group that 3 months after HSCT was (4.57 +/- 6.39) x 10(-3). CML28 can be detected 3 months after HSCT in 1 patient with CML-CP and 3 patients with CML-AP or BC. 2 of them with low level (<2 x 10(-2)) survived without relapse, the other 2 case with high level (>2 x 10(-2)) relapsed within one year, 1 case died and 1 case received the second time HSCT, CML28 level decreased rapidly after HSCT, but still higher than 2 x 10(-2) and relapse has taken place. The conclusions was made that CML28 mRNA level is obviously correlated with the development of leukemia. Serial quantification of CML28 mRNA levels are necessary for HSCT recipients, and more informative than a single detection. Using of this assay to evaluate MRD in the patients received HSCT is helpful for prediction of relapse.
Adolescent ; Adult ; Antigens, Neoplasm ; genetics ; Antigens, Surface ; genetics ; Child ; Exoribonucleases ; genetics ; Exosome Multienzyme Ribonuclease Complex ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; genetics ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Organic Chemicals ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; RNA-Binding Proteins ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Time Factors

OBJECTIVETo investigate the role of STK15 in regulating mitosis of gastric cancer cells (MKN45) by gene silencing through RNA interference mechanism.

METHODSRNA interference technique was used to inhibit STK15 expression in MKN45 cells. The expression levels of STK15 mRNA and protein were measured by real-time quantitative RT-PCR and Western blot respectively and cell morphological changes were investigated by reverse microscopy. In addition, cell cycle distribution and cellular proliferation were determined by flow-cytometry and MTT assay respectively. Finally, the mitotic phenotype of MKN45 cells was studied by immunofluorescence staining and confocal microscopy.

RESULTSSilencing of STK15 gene by RNA interference was confirmed by marked decrease of STK15 mRNA and protein levels in the treated MKN45 cells. This silencing correlated with rounding of the cells, decreasing of DNA content in G(2) phase (P < 0.05) and a lowered proliferation index (P < 0.05), along with alterations of mitotic phenotype of MKN45 (P < 0.05).

CONCLUSIONSTK15 gene may play a key role in regulating cellular mitosis and its inhibition by RNA interference leading to mitosis arrest in MKN45 cells.

Adenocarcinoma ; metabolism ; pathology ; Aurora Kinase A ; Aurora Kinases ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA, Neoplasm ; metabolism ; Gene Silencing ; Humans ; Mitosis ; drug effects ; Protein-Serine-Threonine Kinases ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; pharmacology ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo compare and analyze the MRI features of different renal cell carcinoma (RCC) subtypes.

METHODSThe MR images of 81 surgically and pathologically confirmed renal cell carcinomas from 79 patients were reviewed retrospectively. The MR imaging features of lesions in plain scan, the degree and patterns of lesion enhancement (homogeneous, heterogeneous, peripheral), and tumor spreading patterns were analyzed. In order to evaluate the diagnostic validity of differentiating RCC subtypes using signal enhancement, receiver operating characteristic curves (ROC) were generated. The cutoff value of post-contrast signal intensity to noise ratios (SNR) of the tumor parenchyma were also generated in order to differentiate clear cell RCC from other subtypes.

RESULTSOf the 81 lesions, 58 were clear cell carcinomas, 10 chromophobe cell carcinomas, 8 papillary cell carcinomas, and 5 unclassified RCC. All the chromophobe cell subtype tumors showed a homogeneous density (P < 0.05). The clear cell subtype tumors were likely heterogenous, and also showed heterogenous enhancement with mixed signal than other subtypes (P < 0.05). The cutoff value of SNR, which was used to differentiate clear cell subtype from the other subtypes, were 616 (corticomedullary phase), 579 (nephrographic phase) and 278 (excretory phase), retrospectively. The nephrographic phase is the most appropriate for differentiation, with a sensitivity of 62.1%, specificity of 91.3%, positive predictive value of 94.7%, negative predictive value of 48.8% and an accuracy value of 70.3%. No significant difference was found in tumor spreading patterns among all subtypes of RCC.

CONCLUSIONMR imaging features, particularly tumor heterogeneity and degree of enhancement are useful in differentiation of the renal cell carcinoma subtypes, and in choosing an individualized therapy.

Adult ; Aged ; Carcinoma, Renal Cell ; pathology ; Female ; Humans ; Image Enhancement ; methods ; Kidney Neoplasms ; pathology ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Retrospective Studies ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo observe the effect of polo-like kinase 1 (PLK1) gene depletion on mitosis phenotype and elucidate its vital role in gastric cancer cell line (MKN45) mitosis.

METHODSThe PLK1 expression in MKN45 cells was blocked by RNA interference (RNAi), the expression level of PLK1 mRNA and protein were measured by real-time quantitative PCR and Western blot, respectively. The morphological change of microtubules and mitosis phenotype in MKN45 cells were observed by immunofluorescence staining and laser confocal microscopy, the morphological changes of cells were observed by reverse microscopy, the variation of cell cycle distribution was detected by flow-cytometry.

RESULTSAfter RNAi targeting PLK1, PLK1 mRNA and protein level decreased obviously, the cell microtubules became obscure and lost cohesiveness, the mitosis phenotype also varied substantially (P < 0.05), more gastric cancer cells became rounded and showed G(2) phase cell DNA content (P < 0.05).

CONCLUSIONPLK1 gene plays a key role in mitosis and its inhibition can lead to mitosis arrest in MKN45 cells.

Adenocarcinoma ; enzymology ; metabolism ; pathology ; Cell Cycle Proteins ; biosynthesis ; genetics ; Cell Line, Tumor ; G2 Phase ; drug effects ; Humans ; Mitosis ; drug effects ; Protein-Serine-Threonine Kinases ; biosynthesis ; genetics ; Proto-Oncogene Proteins ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; pharmacology ; Stomach Neoplasms ; enzymology ; metabolism ; pathology ; Transfection

OBJECTIVETo investigate cytotoxicity and genotoxicity of benzo(a)pyrene (B(a)P) by 16HBE-CYP1A1 cells which are human bronchial epithelial cell with CYP1A1 transformed.

METHODSExpression of CYP1A1 and mEH of cell models were tested by real-time quantitative polymerase chain reaction. Cells were treated with 0, 1, 5, 10 and 20 micromol/L B(a)P for 24 h. Adverse effects of B(a)P were tested by cytokinesis-block micronucleus (CBMN) cytome assays. Cytotoxicity was assessed by the nuclear division index (NDI), frequency of necrotic and apoptotic cells. Genetic damages were assessed by frequencies of CBMN, nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs).

RESULTSHigh levels of CYP1A1 and mEH were found in 16HBE-CYP1A1 cells (relative mRNA content was 7.8 x 10(-4) and 0.030 respectively). In 16HBE-CYP1A1 cells, NDI were decreased in 1, 5, 10 and 20 micromol/L B(a)P treated groups, 1.92 +/- 0.04, 1.71 +/- 0.01, 1.61 +/- 0.04, and 1.41 +/- 0.01, respectively; and lower than control group (2.08 +/- 0.03). Compared with control group ((82.67 +/- 6.66)%), the binucleated cells ratios were decreased, (76.33 +/- 3.51)%, (66.33 +/- 0.58)%, (51.67 +/- 1.53)% and (39.0 +/- 1.0)% respectively.Necrotic cells ratios were (1.93 +/- 0.42)%, (2.20 +/- 0.53)%, (8.07 +/- 0.90)% and (15.27 +/- 2.80)%, respectively, higher than control group ((0.47 +/- 0.11)%). The differences were significant (F values were 899.94, 303.33, 240.87, P < 0.01). Apoptotic cells were increased at lower groups and decreased to normal at higher groups treated by B(a)P. They were (1.20 +/- 0.53)%, (2.00 +/- 0.20)%, (1.47 +/- 0.12)%, (1.20 +/- 0.00)% and (1.20 +/- 0.00)%, respectively. Analysis on biomarkers of genetic damage, the significant dose-effect relationship were observed in NPBs and NBUDs (F values were 50.23, 121.09, P < 0.01, respectively). Frequencies of NPBs were (4.67 +/- 2.89) per thousand, (7.33 +/- 1.53) per thousand, (10.67 +/- 2.08) per thousand and (11.00 +/- 1.00) per thousand respectively. Frequencies of NBUDs were (2.33 +/- 0.58) per thousand, (4.00 +/- 1.00) per thousand, (5.00 +/- 1.00) per thousand, and (7.67 +/- 1.16) per thousand respectively. However, the dose-relationship of CBMN last only to 10 micromol/L B(a)P treated groups in 16HBE-CYP1A1 cells, and frequencies of CBMN were (8.33 +/- 3.21) per thousand, (14.67 +/- 1.15) per thousand, respectively. Frequency of CBMN was (16.67 +/- 2.88) per thousand in 20 micromol/L B(a)P treated group, lower than 10 micromol/L B(a)P treated group ((17.67 +/- 2.08) per thousand). In 16HBEV control cells, the cytotoxicity was found only in higher B(a)P treated groups and frequencies of CBMN, NPBs and NBUDs were increased also. While no significant differences were observed between 5, 10, 20 micromol/L B(a)P treated groups (they were (6.37 +/- 2.08) per thousand, (9.33 +/- 1.52) per thousand, (9.33 +/- 3.21) per thousand; (4.33 +/- 1.53) per thousand, (6.00 +/- 2.65) per thousand, (5.33 +/- 1.53) per thousand and (2.33 +/- 0.58) per thousand, (3.33 +/- 1.16) per thousand, (3.67 +/- 1.16) per thousand, respectively).

CONCLUSIONSThe genetic damages were more severe after treated with activated B(a)P, which may be induced by decreased NDI, increased necrotic cells and inhibition of apoptosis.

Apoptosis ; drug effects ; Benzo(a)pyrene ; toxicity ; Cell Division ; drug effects ; Cell Line, Transformed ; DNA Damage ; Humans ; Micronuclei, Chromosome-Defective

OBJECTIVETo observe the effect of polo like kinase 1 (plk1) gene depletion on the growth of gastric cancer cell line-MKN45 cells in vitro and vivo and discuss the feasibility and effectiveness of arranging plk1 as gene therapeutic target for gastric cancer.

METHODSThe plk1 expression of MKN45 cells was inhibited by RNA interference (RNAi). The plk1 mRNA and protein level were measured by real-time quantitative PCR and western blotting, and the change of cell cycle distribution and apoptosis rate were detected by flow-cytometry, and the MKN45 cells proliferation was measured by MTT method. MKN45 cells treated with plk1 siRNA were transplanted subcutaneously in nude mice and their tumorgenesis ability were observed, the plk1 protein levels of the samples from nude mice in different groups were compared.

RESULTSAfter treatment with plk1 siRNA, plk1 mRNA and protein level decreased obviously in certain time, more MKN45 cells accumulated at G(2)/M (P < 0.05). Apoptosis rate of MKN45 cells treated with plk1 siRNA was higher than that of control cells at 48 h and 72 h (P < 0.05), and MKN45 cells proliferated slowly than control groups (P < 0.05), while the tumorgenesis ability obviously decreased, but the plk1 protein levels of the samples from nude mice in different groups were not different.

CONCLUSIONSsiRNA targeting plk1 can inhibit the proliferation of MKN45 cells in vitro and vivo. Plk1 may be a novel therapeutic target for gastric cancer.

Animals ; Apoptosis ; drug effects ; Cell Cycle Proteins ; drug effects ; genetics ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Male ; Mice ; Mice, Nude ; Protein-Serine-Threonine Kinases ; drug effects ; genetics ; Proto-Oncogene Proteins ; drug effects ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; pharmacology ; Stomach Neoplasms ; drug therapy ; enzymology ; pathology ; Transfection