To analyse and compare the advantages and disadvantages of various kinds of examinations in teaching molecular biology,and detail the process and effect of thesis proseminar domained examination.

We studied the effect of photodynamic therapy with phycobiliproteins on human liver cancer cells in vitro. With 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT assay), we used two phycobiliproteins, R-phycoerythrin (R-PE) and C-phycocyanin (C-PC) prepared from Porphyra yezoensis, to determine the killing rates and apoptosis rates of human liver cancer cells (SMMC-7721) mediated by laser. When the concentration of R-PE was 120 mg/L, the survival rate of human liver cancer cells was 27% after treated by Argon laser with 100 J/cm2 doses, while the survival rate in the control group (without adding R-PE) was 65%. When the C-PC concentration was 120 mg/L, the survival cell rate was 47% after treated by He-Ne laser with 35 J/cm2 dose, while the survival rate in the control group (without adding C-PC) was 70%. After handled only with these two kinds of phycobiliproteins for 72 h, the growth of cancer cells presented significant inhibition. The maximal inhibition rates reached up to 31% with R-PE (120 mg/L concentration) and 27% with C-PC (250 mg/L concentration) respectively. After irradiated by laser for 8 h, the maximal cell apoptosis rates were 31.54% with R-PE and 32.54% with C-PC, respectively. It indicated that R-PE and C-PC extracted from Porphyra yezoensis could develop to new photosensitizers for cancer photodynamic therapy.
Apoptosis ; drug effects ; radiation effects ; Cell Line, Tumor ; Humans ; Lasers ; Liver Neoplasms ; pathology ; Photochemical Processes ; Photochemotherapy ; methods ; Phycobiliproteins ; isolation & purification ; pharmacology ; Phycoerythrin ; isolation & purification ; pharmacology ; Porphyra ; chemistry

Objective To evaluate the comparability of test results of self-built human carcinoembryonic antigen(CEA)electro-chemiluminescence immunoassay(ECLIA)and imported reagent.Methods A total of different concentrations 77 fresh serum speci-mens were collected and detected CEA by two kinds of ECLIA kit.The results were analyzed with Excel2003 and SPSS1 9.0 soft-ware.Results The difference between each dose was significant (P <0.05),and the detection results between each had no signifi-cant difference (P >0.05);the sensitivity of the assay was 0.3 ng/mL,the intra coefficient of variation was 4.58%-5.83%,the inter coefficient of variation was 5.07%-5.97%,the analytical recovery was 99.13%-107.28%,the specificity of the assay had no cross reaction with CA1 99 and AFP.The correlation coefficient between two kinds of reagents determination results was greater than 0.95,with imported reagent as reference test,self-built carcinoembryonic antigen ECLIA clinical performance evaluation was acceptable.Conclusion The precision of the two kinds of ECLIA in detection of CEA accord to clinical requirement.Comparability exists in evaluating the acceptability of clinical.

A new analytical method has been developed for the simultaneous determination of CrⅢand CrⅥusing on-line sample pretreatment valve-switching ion chromatography. The organic matrix in leather was removed by using a reverse-phase column as the pretreatment column. Before injection, EDTA was added into sample solution to react with the CrⅢto form anion which could absorb visible light strongly. After injection, the ions separated by the pretreatment column were received in a collection loop. Then the ions were delivered into an analytical column and separated. CrⅥ then was derived with the derivatization reagent 1, 5-diphenylcarbazide ( DPC) , and detected together with CrⅢ-EDTA complex by a UV-Vis detector. Under the optimum conditions, the linear range of the method for CrⅢ and CrⅥ was 0. 3-10 mg/L (r=0. 9991) and 0. 05-2 mg/L ( r = 0. 9992 ), whereas detection limits ( S/N = 3 ) were 80. 78 μg/L and 6. 67 μg/L, respectively. The recoveries were in the range of 88. 7%-108. 5% with the relative standard deviations for retention time and peak area less than 3%. The method could be applied to determine CrⅢ and CrⅥ in leather and cloth effectively and quickly.

We developed large-scale preparation of phycoerythrin from Porphyra haitanensis, a main economic red algae in China. Firstly, P. haitanensis thallus was broken by using "swelling and smash" method. Then times of grads ammonium sulfate precipitation applied to the crude extraction were compared. Desalted solution was further purified with one-step chromatography using hydroxyapatite and properties on spectrum and molecular weight were identified finally. The results indicated that after four times of ammonium sulfate precipitation (15%, 50%, 10% and 40%), the absorption spectrum purity of P. haitanensis achieved 0.9 (A564/A280), and 507.82 mg phycoerythrin (A564/A280 > 3.2) was obtained from 7 kg fresh algae after further hydroxyapatite chromatography. This research provides a potential way for preparation of phycoerythrin in large sclae.
Ammonium Sulfate ; chemistry ; Chromatography ; methods ; Phycoerythrin ; isolation & purification ; Porphyra ; chemistry

Fluorescent proteins can be used as probes to investigate intercellular molecular interactions and trace the pathway of specific metabolites, thus providing a detailed and accurate description of various metabolic processes and cellular pathways in living cells. Nowadays, the existing fluorescent proteins cover almost all spectral bands from ultraviolet to far-red. These fluorescent proteins have been applied in many fields of bioscience with the help of high-resolution microscopy, making great contributions to the development of biology. It is generally agreed that orange fluorescent proteins refer to the fluorescent proteins at the spectral range of 540-570 nm. In recent years, researches on orange fluorescent proteins have made great progress, and they have been widely applied in the field of biology and medicine as reporter protein and fluorescence resonance energy transfer as fluorescent receptor. This paper reviews the studies in the field of orange fluorescent proteins over the last 15 years, with the special focus on the development and application of orange fluorescent proteins to provide the basis for the future studies.
Biosensing Techniques ; trends ; Fluorescence Resonance Energy Transfer ; Luminescent Proteins ; metabolism ; Research ; trends

Macrobrachium nipponensis is delicious and has high economic value, but its susceptibility to white-spot syndrome virus (WSSV) is unknown. Susceptibility, morbidity, and multiplication of WSSV in M. nipponense were studied by epidemiological survey, infection experiment and qPCR. M. nipponense was the natural host of WSSV, and the natural carrying rate was about 8.33%. M. nipponense could be infected with WSSV via oral administration, muscle injection and immersion, and the cumulative infection rate of 10 d exposure was 100%, and the cumulative mortality rates were 100%, 75% and 0%, respectively. The infection of WSSV is fast by muscle injection. The virus content after 5 day's injection is 1 000 times higher than that of the first day of infection, and the mortality rate reached 100% after 8 days. The median lethal dose (LD₅₀) measured as the mortality of infected M. nipponense via injection indicated the LD₅₀ in the concentration of WSSV of 2.71×10⁵ virions/μL. In shrimp farming, M. nipponense can be infected by ingesting WSSV infected shrimp or dead shrimp, and also by soaking in WSSV-containing water and thus become a vector, consequently affecting the spread and pathogenicity of WSSV.

Light quality can regulate both psbA genes and vector promoter psbA of the engineered Synechococcus. Through light regulation, we tried to improve yield of the recombinant protein for vp28 gene-expressed Synechococcus sp. PCC7002. To drive photon-capturing efficiently, three limiting factors (irradiance, temperature and pH) were optimized by measuring net photosynthesis. High cell density cultures were performed with variant ratios of white, red and blue light in a 5-L photo-bioreactor. Yields of biomass, expressions of vp28 and transcription levels of psbA were compared. High ratio blue light-induced vp28 transcription had tripled and the relative accumulation of VP28 protein was doubled. The relative expressions of psbAII and psbAIII had positive correlations with higher ratio of blue light, not the red light. With high ratio red light inducing, dry biomass reached 1.5 g/L in three days. Therefore, we speculated that red light accelerated biomass accumulation of the transgenic strain and blue light promoted transcription for PpsbA and psbA. These results provided useful information for mass production of cyanobacteria and its secondary metabolites.
Gene Expression Regulation, Bacterial ; Light ; Photosystem II Protein Complex ; genetics ; Promoter Regions, Genetic ; Synechococcus ; genetics ; growth & development ; radiation effects