China ; Occupational Health Services ; methods ; organization & administration

To investigate the upregulated genes associated with cold acclimation, a cold acclimation model was established based on Balb/C mouse. mRNA of muscle and liver were isolated, and the upregulated genes of these tissues were studied by representational differential analysis (RDA). The upregulated genes then were sequenced and searched by Blast software in GenBank database. The results showed that some genes were upregulated and possibly associated with cold acclimation. Three of these genes, transferrin, fibrinogen B-beta-chains and a new gene fragment (Genbank ID: AF454762), were confirmed to be upregulated by RNA slot-blot analysis. The finding of these genes might contribute to further understanding of the molecular mechanisms of cold acclimation.
Acclimatization ; genetics ; Animals ; Cold Temperature ; Gene Expression ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Muscle, Skeletal ; metabolism ; Transcriptome ; Up-Regulation

OBJECTIVETo construct a lentiviral expression vector for RNA interference of human CDH22 gene, and assess its gene silencing effect in colorectal cancer cells to provide a basis for investigating the role of CDH22 gene in the signaling pathway involved in human colorectal carcinoma metastasis.

METHODSHuman CDH22 gene short hairpin RNA (shRNA) sequence was designed using a software available on-line. After synthesis and annealing, the double-stranded oligonucleotides (dsOligoe) were cloned into the pENTR(TM)/U6 plasmid followed by sequence analysis. A positive clone was subcloned into pLenti6/BLOCK-iT(TM)-DEST vector and transformed into stb13 competent cells, with also verification by sequencing. The recombinant lentivirus was harvested from 293FT cells contransfected with the positive recombined plasmid and lentiviral packing materials. SW480 cells were infected with the recombinant lentivirus and the cells with stable CDH22 knock-down were screened by blasticidin selection. CDH22 expression in the cells was determined by real-time reverse transcription-polymerase chain reaction.

RESULTSA recombinant lentiviral vector expressing shRNAs against CDH22 gene was obtained and confirmed by DNA sequencing. Fifteen clones of SW480 cells infected with the recombinant lentivirus were selected, and clone 11 exhibited substantial knock-down of CDH22 mRNA expression.

CONCLUSIONThe lentiviral shRNA expression vector targeting human CDH22 gene capable of stable CDH22 gene knock-down in SW480 cells has been successfully constructed, which provides a basis for further study of the relationship between human colorectal carcinoma and CDH22 gene.

Base Sequence ; Cadherins ; biosynthesis ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo identify the region in PRL-3 gene promoter where the transcriptional factor Snail can bind.

METHODSPRL-3 promoter and the possible binding sites of the transcription factors were analyzed by bioinformatical methods. Chromatin immunoprecipitation and PCR were performed using the antibody specific for Snail to verify the binding of Snail to PRL-3 promoter.

RESULTSAccording to the prediction by TRED, a promoter prediction software, PRL-3 gene promoter was located between -700 bp to 299 bp of PRL-3 gene. Many possible transcription factor binding sites such as for Snail, n-MYC, ARNT, E74A, NF-kappaB, NRF-2 and AML-1 were predicted by Consite, a promoter analysis web system. Interestingly, a 5'-CACCTG-3' core sequence and other related sequences of Snail binding sites were found in the promoter region of PRL-3 genes by Consite software. Two regions in PRL-3 promoter were validated to allow binding of Snail by chromatin immunoprecipitation analysis of SW480 cells.

CONCLUSIONSSnail regulates the activity of PRL-3 gene by binding to the promoter of PRL-3 gene in SW480 cells.

Base Sequence ; Binding Sites ; Cell Line, Tumor ; Computational Biology ; Humans ; Molecular Sequence Data ; Neoplasm Proteins ; metabolism ; Promoter Regions, Genetic ; Protein Tyrosine Phosphatases ; metabolism ; Snail Family Transcription Factors ; Software ; Transcription Factors ; metabolism

OBJECTIVETo develop a new stable and efficient reagent for methemoglobin reduction test.

METHODSThe results of methemoglobin reduction test using the new reagent were compared with those by G6P/6PG ratio method and classic methemoglobin reduction test.

RESULTSThe new reagent was stable for at least 6 months at room temperature and 12 months at 2-8 degrees celsius;. The results of the test using this new reagent were stable and reliable.

CONCLUSIONThe new reagent for methemoglobin reduction test allows easy operation with well reproducible results and can be used in clinical screening of glucose-6-phosphate dehydrogenase deficiency.

Glucosephosphate Dehydrogenase ; analysis ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; Humans ; Mass Screening ; Methemoglobin ; metabolism ; Oxidation-Reduction ; Reagent Kits, Diagnostic ; Reproducibility of Results

OBJECTIVETo detect the correlation between the expression of topoisomerase 2 alpha (TOP2A) and the biological behaviors of human colorectal carcinoma.

METHODSImmunohistochemistry and real-time RT-PCR were used to detect the expression of TOP2A in colorectal carcinomas and normal mucosa.

RESULTSThe protein and mRNA expressions of TOP2A in the metastatic lymph nodes were significantly higher than those in matched primary lesions and normal tissues (P<0.05). No significant difference was found in TOP2A expressions between normal mucosa and colorectal carcinomas. The protein and mRNA expressions of TOP2A were significantly correlated to the lymph node metastasis and invasion depth (P<0.05), but not to the differentiation of the tumor (P>0.05).

CONCLUSIONTOP2A plays an important role in the invasion and metastasis of the colorectal carcinomas, and may serve as a valuable indicator for the diagnosis, treatment and the prognostic evaluation of the malignancy.

Antigens, Neoplasm ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; DNA Topoisomerases, Type II ; metabolism ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Male ; Poly-ADP-Ribose Binding Proteins

Objective To observe the efficacy and safety of applying sirolimus-eluting stents in emergency percutaneous coronary intervention (PCI) for the patients with acute myocardial infarction (AMI).Methods In total,220 patients with AMI were enrolled in this study at Shanghai Tongji Hospital, divided into two groups,one with bare-metal stent and the other with sirolimus-eluting stent.Cardiovascular fatality,major adverse cardiac events (MACE) and target vessel revascularization (TVR) were observed one and six months after PCI in the two groups.Results There was no significant difference in overall fatality and MACE in the 1~(st) or 6~(th) months after PCI between the two groups.Three cardiogenic deaths occurred in bare-metal stent group with a fatality of 2.8 percent,and five deaths in sirolimus-eluting stent group with a fatality of 4.5 percent in six months after PCI.However,rate of restenosis in those with sirolimus-eluting stents was significantly lower than that of bare-metal stents (6.0 percent vs 16.1 percent,P

OBJECTIVETo carry out epidemiological study on an outbreak caused by E. coli O157:H7 infection in Jiangsu province in 1999.

METHODSEpidemiological, microbiological and moleculebiological methods were used to find out the source, route of transmission and risk factors.

RESULTS95 severe O157:H7 infected patients with acute renal failure in 9 counties and districts of 2 municipalities were reported in Jiangsu province, 1999 while 83 of the patients died with a death rate of 87.37%. Most patients were seen in mid or late June. The ratio of male to female was 1 to 1.44 and 88.42% of the patients were over 50 years old. 38 patients occurred in 2000 with 34 deaths. Major factors contributing to the outbreak would include without drinking tap water, eating leftover food, poor sanitary status in kitchen, not washing hands before meal and after bowl movement. 2 strain of O157:H7 was isolated from severe patients and 3 from diarrhea cases. Carrier rate among animals was up to 9.62% and 99.41% of the strains carried toxic gene. Strains isolated from feces of patients and animals belonged to the same colonies.

CONCLUSIONThis outbreak was severe which caused by O157:H7 and was first seen in China, which was closely related to the high carrier rate of O157:H7 in animals and to the positive rate of high toxic gene of the strains. There were various routes of transmission and the main factors of infection would include poor personal health habits and poor sanitation of the household.

Acute Kidney Injury ; epidemiology ; etiology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Bacterial ; immunology ; Case-Control Studies ; Child ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; microbiology ; Disease Outbreaks ; Escherichia coli Infections ; complications ; epidemiology ; Escherichia coli O157 ; isolation & purification ; Escherichia coli Proteins ; immunology ; Female ; Hemolysin Proteins ; immunology ; Humans ; Infant ; Male ; Middle Aged ; Seroepidemiologic Studies

OBJECTIVETo study expression of the zinc finger transcriptional factor Snail in colorectal carcinoma and its significance.

METHODSExpressions of Snail in colorectal carcinoma SW480 and SW620 cells were assayed by immunocytochemistry and immunofluorescent cytochemistry. The paraffin-embedded specimens from 68 cases of colorectal carcinoma and its corresponding adjacent tissues, 33 cases of adenoma and 35 cases of metastatic lymph nodes were also examined for Snail expressions using immunohistochemistry.

RESULTSSnail protein was located mainly in the cell nucleus of SW480 and SW620 cells. The expressions of PRL-3 protein in the specimens of colorectal carcinoma, normal mucosa, adenoma and metastatic lymph nodes were significantly different (Chi(2)=92.852, P=0.000). In the adenoma tissues, the expression was significantly higher than that in normal mucosa (Z=-2.902, P=0.004), the metastatic lymphnodes had significantly higher expressions than the primary colorectal carcinomas (Z=-4.951, P=0.000), which, in turn, showed significantly higher expression than the adenoma tissues (Z=-3.572, P=0.000). Significant correlation of Snail expression was found to the progression and metastasis of colorectal carcinoma (Z=-2.043, P=0.041).

CONCLUSIONThe expression of Snail is significantly correlated to genesis, progression and metastasis of colorectal carcinoma.

Cell Line, Tumor ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Snail Family Transcription Factors ; Transcription Factors ; metabolism ; Zinc Fingers

OBJECTIVETo study the relevant position of the pedicle of C1 to the lateral mass of C(2-4), set up an identification technique for the entry point decision of C1 pedicle screw by using the lateral mass of C(2-4) as anatomic landmarks.

METHODSTwenty cadaver specimens were used to measure the distance from the sagittal midline of spine to the medial border, the midpoint and the lateral border of C1 pedicle or the lateral mass of C2, C3 or C4. The anatomic relation between the measurements data of C1 pedicle and that of the lateral masses of the cervical vertebrae were analyzed, and the technique of C1 pedicle screw fixation was established.

RESULTSThe average medial border of the lateral mass of C2, C3 and C4 was 0.37 mm, 0.27 mm and 0.24 mm lateral to that of C1 pedicle, the average midpoint of the lateral mass of C2, C3 and C4 was 1.18 mm, 1.41 mm and 1.74 mm lateral to that of C1 pedicle, and the average lateral border of the lateral mass of C2, C3 and C4 was 1.96 mm, 2.54 mm and 3.24 mm lateral to that of C1 pedicle, respectively.

CONCLUSIONThere is a steady anatomic location relation between C1 pedicle and the lateral mass of C2, C3 or C4. As well as the lateral mass of C2, the lateral mass of C3 or that of C4 could be convenient anatomic landmarks to determine the location of C1 pedicle and the position of C1 pedicle screw entry point.

Adult ; Cadaver ; Cervical Atlas ; anatomy & histology ; surgery ; Cervical Vertebrae ; anatomy & histology ; surgery ; Female ; Humans ; Male ; Spinal Fusion ; methods