Student preparation for laboratory sessions is the first step of conducting laboratory experiments. It makes students maximize use of laboratory time and efficiently perform laboratory exercises in open labs. In view of teaching features and requirements of Microbiology experiment, we designed and developed ‘Online Student Preparation and Management System of Microbiology Experiment’, which integrated func- tions of student preparation for laboratory sessions and teacher management. In the system each experiment consists of six successive parts, viz., learning objectives, principle, materials and equipments, procedure video, manipulation simulation and online quiz. Teaching practices showed that the application of the system enhanced the preparing quality and makes the management of the experiment teaching more normalized and efficient. It was an effective measure in improving experimental teaching of Microbiology.

Selective markers used in yeast vector for gene manipulation were usually drug resistance or autotrophy. Unfortunately, drug resistance selective marker requires drug sensitive host and most industrial strains were not autotrophy. In this paper, flocculation gene (FLO1) from Saccharomyces cerevisiae ABXL-1D was amplified by PCR, sequenced and cloned to construct an expression vector. The new vector was easy to manipulate and suitable for broad host of yeasts without either autotrophy or drugs. beta-glucosidase gene from Bacillus polymyxa was cloned with the vector and expressed in Saccharomyces cerevisiae. The specific activity of beta-glucosidase of the recombinant yeast cell-free extract was 3.91 u/mg protein. The residue glucose of the recombinant yeast was considerably reduced in mixed fermentation of glucose and cellobiose. It should be favorable for ethanol fermentation when utilize lignocellulosic biomass as raw material.
Bacillus ; enzymology ; genetics ; Flocculation ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Saccharomyces cerevisiae ; genetics ; metabolism ; beta-Glucosidase ; biosynthesis ; genetics

Objective To study the academic level of randomized control trials(RCTs)published from 2004 to 2009 in Chinese Journal of Hepatobiliary Surgery.Methods Published work about RCTs in 72 issues of Chinese Journal of Hepatobiliary Surgery was searched by finding relevant articles published during 2004-2009.Correlated indexes including literature received time, literature publishing time, the total number of citations, number of citations in Chinese, number of citations in English, research funding situations, the number of authors, article affiliations in all RCTs, together with single(or multiple)center study situation, positive conclusions, the starting time, the end time,and number of patients recruited in clinical RCTs were extracted for aggregate analysis.Results During the past six years, a total of 135 articles of RCT were published in Chinese Journal of Hepatobiliary Surgery, accounting for 8.7% in all clinical studies.The average number of RCTs in each issue was 1.88.The average time for publishing time delay was 450 days.The total number of citations in RCTs was 1252 and the average number of RCTs for citations was 9.3.53 RCT studies were financed, accounting for 39% of the total number of research papers.Beijing Friendship Hospital attached to the Capital University of Medical Sciences had published 8 papers of RCT and it is the institution of publishing the largest number of such papers in the journal.The number of multi-center study for RCTs was one and all RCT studies had significant conclusions.The average research duraton in RCTs was 45 months.The total number of patients in 33 clinical RCTs was 2705 and the average number of patients recruited in each RCT was 82.Conclusion Chinese Journal of Hepatobiliary Surgery put emphasis on the strongest evidence and the most valuable RCT researches that have great significance in the field of domestic hepatobiliary surgery for clinical treatment and experimental research.

Bronchoscopy has been an important tool for clinicians. Based on the traditional training, this research tries to introduce microteaching combined with simulated bronchoscope, strengthens the chest CT reading training, standardizes operation practices, which significantly improve the operation speed, flex-ibility and accuracy, shorten the training cycle of the novices. This medical teaching method are effective to contribute the novices to master the operation skill of bronchoscopy.

OBJECTIVETo explore GAGA-like element binding protein in human cells.

METHODSYeast one-hybrid system was used to screen the GAGA-like element binding proteins in HTLV-1 transformed Jurkat cell cDNA fusion library. Total RNA extracted from Jurkat cells was first labeled by reverse transcription, and was taken as cDNA probe to hybridize with the candidate positive clones.

RESULTS9 positive clones were obtained, and 6 out of the 9 clones were positively hybridized with the cDNA probe.

CONCLUSIONS6 candidate clones encoding for GAGA-like element binding proteins were obtained from Jurkat cells for further investigation.

Carrier Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA-Binding Proteins ; Drosophila Proteins ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Homeodomain Proteins ; genetics ; Human T-lymphotropic virus 1 ; genetics ; Humans ; Jurkat Cells ; Leukemia, T-Cell ; metabolism ; pathology ; Transcription Factors ; genetics ; Two-Hybrid System Techniques

OBJECTIVETo study the effect of Rac-MEKK-JNK (Rac-mitogen activated protein kinase kinase kinase-C-jun N-terminal protein kinase) signal pathway on heat shock-induced hsp90 beta gene expression and the impact of Hsp90 on the regulation of the pathway.

METHODSDN-Rac, DN-MEKK or DN-JNK were cotransfected with hsp90 beta CAT reporter plasmid beta 3.1 into Jurkat or LETPa-2 cells individually, the CAT mRNA expression was then determined quantitatively by competitive RT-PCR based system. Western blot was carried out to detect the expression level and phosphorylation of c-Jun in Jurkat and LETPa-2 cells that were transfected with DN-Rac, DN-MEKK or DN-JNK. By in vitro kinase activity assay and Western blot, the effect of geldnamycin (GA) on heat induced JNK activity were evaluated.

RESULTSIn Jurkat cell transfected with DN-Rac, DN-MEKK or DN-JNK, heat shock induced relative CAT mRNA expression level was decreased to (72.8 +/- 5)%, (60 +/- 13.2)% and (47.7 +/- 12.1)% of the control respectively; while in LETPa-2 cell hsp90 beta 3.1 reporter gene expression was accordingly suppressed to (16.17 +/- 5.1)%, (50.2 +/- 8.7)% and (47.5 +/- 10)% of control. C-Jun expression and phosphorylation were inhibited by the transfection of either one of DN-Rac, DN-MEKK or DN-JNK. With GA treatment, heat shock induced JNK activity was repressed, while the expression level of JNK or c-Jun was not obviously changed.

CONCLUSIONSRac-MEKK-JNK pathway promotes heat shock induced hsp90 beta gene expression and hsp90 may participate in the regulation of heat shock activated Rac-MEKK-JNK signal pathway in both Jurkat and LETPa-2 cells.

Benzoquinones ; Cell Line, Tumor ; Genes, Reporter ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Hot Temperature ; Humans ; JNK Mitogen-Activated Protein Kinases ; Lactams, Macrocyclic ; Leukemia, T-Cell ; pathology ; Mitogen-Activated Protein Kinase Kinases ; physiology ; Mitogen-Activated Protein Kinases ; physiology ; Protein Kinase C ; physiology ; Quinones ; pharmacology ; Signal Transduction ; Transfection

OBJECTIVENan'ao County in Guandong Province is a high-risk area of esophageal cancer in Southern China. Of the suspected etiological factors in the environment, N-nitrosamines and their precursors have received the greatest attention.

METHODSSixty samples of the diet ingested by the inhabitants were collected and detected for volatile N-nitrosamines and their precursors. Five N-nitrosamines detected by Gas Chromatography-Thermal Energy Analyzer were N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosopiperidine and N-nitrosomethyl-benzylamine.

RESULTSThe average content of 5 volatile N-nitrosamines in the diet was 312.0 micrograms/kg (median). The daily intake of the nitrosamines was 286.5 micrograms/head/day. Only the ability to exogenously synthesize N-nitrosopiperidine was powerful among 5 volatile N-nitrosamines. By a computerized stepwise regression analysis and curve fitting, we studied the correlation among the nitrosamines, the precursors and the major food items in the samples.

CONCLUSIONIt demonstrated that a relatively high content of volatile N-nitrosamines was present in the diet collected in the area.

Adult ; China ; epidemiology ; Diet ; Esophageal Neoplasms ; epidemiology ; etiology ; Female ; Humans ; Male ; Middle Aged ; Nitrosamines ; administration & dosage ; adverse effects ; Risk Factors ; Volatilization

One pair of degenerate primer was designed according to conserved motifs of the psaB (A2 subunit of photosystem I) of Chlamydomonas reinhardtii, Chlamydomonas moewusii, Chlorella vulgaris and Mesostigma viride, and a total RNA of Dunaliella salina (D. salina) was extracted with TRIzol reagent. A cDNA fragment, about 1.8kb in length, from green algal D. salina was obtained through RT-PCR method. The resulting PCR product was cloned into T-vector and screened to determine its sequence. Homologous analysis of the deduced amino acid sequence was performed by BLAST and subsequeqtly compared with GenBank data. The obtained cDNA sequence was 1815 bp long, which encodes 605 amino acids (GenBank accession number: AY820754). The sequence shared high homologue with the following psaB: Chlamydomonas reinhardtii 92%, Chlamydomonas moewusii 91%, Chlorella vulgaris 86%, Mesostigma viride 85%, Physcomitrella patens subsp. Patens 85% and Nephroselmis olivacea 84%. It can be concluded that the cloned sequence is psaB cDNA fragment from D. salina.
Algal Proteins ; genetics ; Amino Acid Sequence ; Animals ; Chlamydomonas reinhardtii ; genetics ; Chlorophyta ; genetics ; metabolism ; Cloning, Molecular ; DNA, Complementary ; genetics ; Molecular Sequence Data ; Photosystem I Protein Complex ; genetics ; Sequence Analysis, Protein ; Sequence Homology, Amino Acid

OBJECTIVETo study the effect of Chinese herbal compound (CHC) on the expression of hepatocyte cytochrome P450IIE1 in rat model of alcoholic fatty liver (AFL).

METHODSThe AFL rats models were established by administering the drinking water with 40%(v/v) ethanol, and the changes of pathology in liver and hepatocyte P450IIE1 expression, as well as the contents of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), vitamin E (VitE) in liver were detected and compared with those in the control group.

RESULTSFatty degeneration in liver recovered normally in the CHC-treated group. Immunohistochemical and in situ hybridization examination showed that CHC could inhibit the hepatocyte cytochrome P450IIE1 expression markedly, and restore the contents of MDA, SOD, GSH, VitE to nearly normal range.

CONCLUSIONCHC can prevent AFL through inhibiting the hepatocyte cytochrome P450IIE1 expression markedly

Animals ; Cytochrome P-450 CYP2E1 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fatty Liver, Alcoholic ; pathology ; Gene Expression ; Hepatocytes ; drug effects ; enzymology ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of Inonotus obliquus polysaccharides on testicular injury induced by exposure to high power microwave (HPM) in rats.

METHODSA total of 30 male Wistar rats were randomly divided into 5 groups, i.e., the normal control group, the microwave radiation model group, the treatment group, the new microwave radiation model group, and the prevention group, 6 in each group. All rats, except those in the normal control group, were exposed to microwave at an average power density of 200 mW/cm2 for 6 min. Rats in the control group and the model group were administered with normal saline by gastrogavage, once a day. Rats in the treatment group and the prevention group were given with Inonotus obliquus polysaccharides by gastrogavage, 2 mL each time (400 mg/kg body weight), once a day. All rats were sacrificed on the 11th day.The sperm density and the rate of sperm deformity were determined. Pathological changes of testis were observed by light microscope and transmission electron microscope.

RESULTSShort-term HPM irradiation could significantly reduce the sperm density and increase the sperm deformity rate (P < 0.05). Meanwhile, obvious pathological changes of testes occurred. Compared with the two model groups, the sperm density increased and the sperm deformity rate decreased in the treatment group and the prevention group (P < 0.05). Under the light microscope, injuries of spermatogenic cells and stromal cells, as well as vascular dilatation and congestion were obviously alleviated in the treatment group and the prevention group. Mitochondrial swelling and endoplasmic reticulum expansion shown by ultrastructural observation were also significantly alleviated. Of them, injuries of spermatogenic cells and inflammation response were milder in the treatment group than in the prevention group.

CONCLUSIONSInonotus obliquus polysaccharides had significant protective effect on microwave radiation induced testicular injury. Better effect was obtained by therapeutic medication than preventive medication.

Animals ; Basidiomycota ; chemistry ; Male ; Microwaves ; adverse effects ; Polysaccharides ; pharmacology ; Radiation Injuries, Experimental ; prevention & control ; Radiation-Protective Agents ; pharmacology ; Rats ; Rats, Wistar ; Testis ; drug effects ; pathology ; radiation effects