Candida shehatae xyl1 gene and Pichia stipitis xyl2 gene were amplified by PCR and the xyl1 and xyl2 were both placed under the promoter GAL of vector pYES2 to produce the recombinant expression vector pYES2-P12. Subsequently the pYES2-P12 vector was transformed into S. cerevisiae YS58 by LiAc to produce the recombinant yeast YSS8-12. It was indicate that the recombinant yeast YSS8-12 could converse xylose to ethanol with the xylose consumption rate of 81. 3%.

Animals ; Cell Proliferation ; Hepatic Stellate Cells ; Liver Cirrhosis/metabolism* ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; Rats

OBJECTIVETo establish a novel suspension microarray technology for the detection of three kinds of veterinary drug residues: chloramphenicol, clenbuterol and 17-beta-estradiol (CAP, CL and E2).

METHODSThe three conjugates that veterinary drug coupled with bovine serum albumin (BSA) were synthesized and identified by ultraviolet (UV) spectrophotometry and mass spectrum. The veterinary drug conjugates were immobilized on the polystyrene fluorescent microspheres/beads. There were competitive reactions between the veterinary drugs in the aqueous phase and that on the beads for combination with their specific biotinylated monoclonal antibodies. The optimum amount of the veterinary drug conjugates and the antibodies were optimized and selected. The detective standard curves were plotted. The specificity and the unknown samples were also determined by grouping according to different concentrations of the interferes and the samples. Meantime, the different microstructures of the surfaces of the beads were also observed by scanning electron microscope.

RESULTSCouplings were completed between small molecular veterinary drugs and BSA. The amounts of the three conjugates and the antibodies were optimized. The detective standard curves of the suspension array and their corresponding coefficients of determination (R2) were good (R2 > 0.99). The detection ranges of the three veterinary drugs were (40.00 - 6.25) x 10(5) ng/L, (50.00-7.81) x 10(5) ng/L and 1.00 x 10(3) - 7.29 x 10(5) ng/L respectively. Simultaneously, the specific detection of the suspension microarray was excellent and did not indicate significant cross-reactions. Errors between the found and the real are in the range of 8.09% - 17.03%. It can be considered that the relative standard deviations were relatively small. Successful couplings were also directly confirmed by the observation for microstructures of the surfaces of the beads by scanning of electron microscope and laid good foundation for the following responses.

CONCLUSIONThe high-throughput suspension microarray should provide a novel method for multi-analysis of the veterinary drugs and have a wide applicative prospects with simple operation, sensitive, rapid and low cost.

Chloramphenicol ; analysis ; Clenbuterol ; analysis ; Drug Residues ; analysis ; Estradiol ; analysis ; Microarray Analysis ; methods ; Veterinary Drugs ; analysis

OBJECTIVETo investigate the roles of Kruppel-like factor 6 (KLF6) and its splice variant KLF6V on suppressing growth and inducing differentiation of human hepatocellular carcinoma hepG2 cells.

METHODKLF6V cDNA was amplificated by RT-PCR from human hepatocellular carcinoma (HCC) tissue and then sequenced. The recombinant vectors expressing KLF6 variant (KLF6V) were constructed using molecular clone technology based on established plasmid pcDNA3.1A(-)/wtKLF6. KLF6V or KLF6-transfected HepG2 cells were established after being screened with G418. Growth activity of HepG2/KLF6 or HepG2/KLF6V cells was detected by in vitro MTT assay. Expression of p21WAF1 or cyclin D1 protein was detected by Western blot, and expressions of AFP or ALB protein were measured by radioimmunoassay.

RESULTSA novel alternatively spliced transcript of the human KLF6 gene was found and its sequencing revealed that the variant form of KLF6 lacked 126nt and its encoded protein products had a deletion of 42 aa near the COOH-terminal amino acid in comparison with full-length KLF6. Although KLF6 alternative splicing was present in both normal and cancerous tissues, expression of the KLF6 splice variants seemed to be up-regulated in HCCs tissues. The isoform of KLF6 proteins antagonized the ability of wild-type KLF6 to up-regulate p21 expression or down-regulate cyclin D1 expression and suppress HepG2 cell proliferation. KLF6 gene increased albumin production and decreased alpha fetoprotein production of the cells.

CONCLUSIONThe isoform of KLF6 protein, present in HCC tissue, antagonizes the ability of wild-type KLF6 to suppress cell proliferation and induce cellular differentiation.

Cell Differentiation ; Cell Proliferation ; DNA, Complementary ; Gene Expression Regulation, Neoplastic ; Hep G2 Cells ; Humans ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; Protein Isoforms ; genetics ; Proto-Oncogene Proteins ; genetics ; Transfection

Carcinoma, Hepatocellular ; chemistry ; Chromatography, High Pressure Liquid ; Female ; Hemoglobins ; analysis ; Humans ; Liver Neoplasms ; chemistry ; Male ; Mass Spectrometry ; Opioid Peptides ; analysis ; Peptide Fragments ; analysis

OBJECTIVETo reveal the relationship among congenital Toxoplasma gondii (T. gondii) infection, T lymphocyte cell subsets in umbilical cord blood and pregnancy outcome.

METHODS784 umbilical cord blood samples were collected and information of pregnancy outcomes was collected in a hospital of Hefei city, Anhui province during March 2009 to May 2010. T. gondii IgM antibodies in the sera were detected by ELISA. For all neonates infected with T. gondii and 10 healthy neonates, T lymphocyte cell subsets were detected by flow cytometry.

RESULTSAccording to the detection results of T. gondii IgM antibodies, 784 neonates were divided into infection group (21 neonates) and control group (763 neonates). The body weight and 1 min Apgar score of infection group were (3116.4 ± 352.6) g and (8.21 ± 1.26) points, respectively, which were statistically lower than control group ((3220.1 ± 242.3) g and (8.77 ± 1.61) points, respectively) (P < 0.01). The proportion of adverse pregnancy outcome of infection group was 19.0% (4/21), which was statistically greater than control group (4.8%, 37/763) (P < 0.01). The percentage of CD(3)(+) T lymphocyte cells in umbilical cord blood in infection group with and without adverse pregnancy outcomes were (64.51 ± 5.27)% and (64.32 ± 4.56)%, respectively, which were statistically lower than control group ((69.32 ± 4.32)%) (P < 0.01). The ratio value of CD(4)(+)/CD(8)(+) in infection group with, without adverse pregnancy outcomes and control group are 1.39 ± 0.24, 1.64 ± 0.28 and 2.34 ± 0.46, respectively, which showed statistical difference between any 2 groups (P < 0.01).

CONCLUSIONT. gondii infection leads to adverse pregnancy outcomes and disorder of cellular immunity while T lymphocyte cell subsets are closely associated with adverse pregnancy outcome.

Case-Control Studies ; Female ; Fetal Blood ; cytology ; immunology ; Humans ; Infant, Newborn ; Pregnancy ; Pregnancy Outcome ; T-Lymphocyte Subsets ; immunology ; Toxoplasmosis, Congenital ; immunology

BACKGROUND AND OBJECTIVERadiotherapy is effective in treating nasopharyngeal carcinoma (NPC). This study evaluated the treatment efficacy, toxicity, and prognostic factors of intensity-modulated radiotherapy (IMRT) in the treatment NPC.

METHODSBetween September 2003 and September 2006, 305 patients with NPC were treated with IMRT in Fujian Provincial Cancer Hospital. IMRT was delivered as follows: gross tumor volume (GTV) received 66.0-69.8 Gy in 30-33 fractions, high-risk clinical target volume (CTV-1) received 60.0-66.65 Gy, low-risk clinical target volume (CTV-2) and clinical target volume of cervical lymph node regions (CTV-N) received 54.0-55.8 Gy. Patients with stages III or IV disease also received cisplatin-based chemotherapy. All patients were assessed for local-regional control, survival, and toxicity.

RESULTSWith a median follow-up of 35 months (range, 5-61 months), there were 16, 8, and 39 patients who had developed local, regional, and distant recurrence, respectively. The 3-year rates of local control, regional control, metastasis-free survival, disease-free survival, and overall survival were 94.3%, 97.7%, 86.1%, 80.3%, and 89.1%, respectively. Multivariate analyses revealed that T-classification had no predictive value for local control and survival, whereas N-classification was a significant prognostic factor for overall survival (P < 0.001), metastasis-free survival (P < 0.001), and disease-free survival (P = 0.003). For stages III-IV disease, concurrent and adjuvant chemotherapy did not influence prognosis. The most severe acute toxicities included Grade III mucositis in 14 patients (4.6%), Grade III skin desquamation in 90 (29.5%), and Grades III-IV leucocytopenia in 20 (6.5%). There were 7% patients with Grade II xerostomia after 2 years of IMRT, no Grades 3 or 4 xerostomia was detected.

CONCLUSIONSIMRT provided favorable locoregional control and survival rates for patients with NPC, even in those with locally advanced disease. The acute and late toxicities were acceptable. N-classification was the main factor of prognosis. Further study is needed on chemotherapy for patients with NPC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Leukopenia ; etiology ; Lymphatic Metastasis ; Male ; Middle Aged ; Mucositis ; etiology ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; radiotherapy ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Radiotherapy Dosage ; Radiotherapy, Intensity-Modulated ; adverse effects ; methods ; Retrospective Studies ; Survival Rate ; Xerostomia ; etiology ; Young Adult

OBJECTIVETo probe blood serum Ab-IgG characteristics of severe acute respiratory syndrome (SARS) patients in Guangzhou and investigate the related factors.

METHODSThe serum of such population diagnosed as SARS convalescent patients, non-SARS patients, family consanguineous contraction persons, wild animal and vegetable salesman and community common people was collected. The lab detective method of ELISA was adopted for these serum samples. And the epidemic investigations for the SARS patients were also carried out.

RESULTSOf these populations, the detective rate of Ab-IgG for the clinic diagnosed SARS patients, which was 53.7%; That for the wild animal salesman and community common people were 16.7% and 0.9%, respectively. Among the clinic diagnosed SARS patients, the positive antibody detective rate was 90.4% for those which had specific contact history or infectivity, which was higher than that for other population. Among the specific contact history or infectivity cases, the antibody positive rate for the young and the old was lower than that for the adult. Meanwhile the difference did not exist among other cases. The antibody positive rate was identical between the male and the female. And the antibody detective rate was decreased by the month.

CONCLUSIONAs a whole SARS-CoV Ab-IgG detective rate for the clinic diagnosed SARS patients was 53.7% only. The reasons for that mainly lie in the wrong clinic diagnosis besides these factors such as age, hormone use and reagent and so on. The combination of lab detection results and epidemic investigation was propitious to the diagnosis veracity. It was impossible for the sub-clinic infection of SARS-CoV virus. The importance in the virus transmitting course need to be further studied.

Adult ; Antibodies, Viral ; blood ; Child ; China ; epidemiology ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; SARS Virus ; immunology ; Seroepidemiologic Studies ; Severe Acute Respiratory Syndrome ; epidemiology ; immunology

Animals ; Cellular Reprogramming ; Dentate Gyrus ; cytology ; Fibroblasts ; cytology ; Mice ; Neural Stem Cells ; cytology ; transplantation ; Swine