The sfa1 gene encoded a bifunctional enzyme with the activities of both alcohol dehydrogenase and glutathione-dependent formaldehyde dehydrogenase in Saccharomyces cerevisiae.The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 nucleotides complementary to sequences in the templates(pUG6 and pUG66 marker plasmid)at 3' end and 45 nucleotides at 5' end that annealed to sites upstream or downstream of the genomic target sequence to be deleted.After two linear disruption cassettes with a Cre/loxP mediated marker were transformed into the cells of Saccharomyces cerevisiae YS-1,the positive transformants were checked by PCR to correct the integration of the cassette and concurrent deletion of the chromosomal target sequence.Once correctly integrated into the genome,the select marker can be efficiently rescued by transformating the plasmid pSH47 into YS-1 and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.The expression of the Cre recombinase finally resulted in the removal of the marker gene,leaving behind a single loxP site at the chromosomal locus.The diploid mutant YS-1-sfa1 was generated,which could enhance the output of ethanol with 8.0% by shaking culture in flask compared with the original strain YS-1.

Gene Library ; Humans ; Nucleic Acid Hybridization ; methods ; Pulmonary Fibrosis ; diagnosis

Objectives To observe the changes of magnetic resonance imaging(MRI) during the course of treatment in hepatolen-ticular degeneration (HLD) and to evaluate its related factors.Methods Sixty - three patients with HLD received routine SE sequence MRI of brain and liver.Twenty - two patients with abnormal MRI in 42 patients and each of them had 2-4 times of serial MRI ex-aminations at interval of 1.0- 1.5 year.Results Ten patients had lesion in both brain and liver in 22 patients. Nine patients only in brain, 3 patients in liver.The major lesions disappear or decrease after rational treatments. There were only slight or no changes of MRI abnormalities in patients receiving treatments or age of patients was over 15 year old. Abnormal MRI findings shown again after stopped treatments over 2 months. In such cases, the clinical and MRI improvements were slow.The rate of clinical and MRI improvement in patients with adequate treatment were markedly higher than patients with inadequate treatment.Conclusions Liver and brain lesion will show in MRI disappear after adequate treatment;There are only slight or no change or showing again in MRI abnormalities in pa-tients with inadequate treatments compared with poor clinical improvement. MRI is beneficial to understanding therapeutic effect and prognosis of HLD.

China ; Humans ; Pneumoconiosis ; diagnostic imaging ; Quality Control ; Radiography, Thoracic ; X-Ray Film

Acoustic Stimulation ; adverse effects ; Animals ; Calcium ; metabolism ; Female ; Hypothalamus ; metabolism ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley

Diagnosis, Differential ; Diarrhea ; etiology ; Humans ; Infant ; Male ; Mediastinal Cyst ; complications ; diagnosis ; surgery ; Mediastinal Neoplasms ; complications ; diagnosis ; Respiratory Sounds ; etiology ; physiopathology ; Treatment Outcome ; X-Rays

Objective To investigate the association of two functional polymorphisms of tryptophan hydroxylase 2(THP2) gene,rs4570625 and rs4565946,with poststroke anxiety disorders.Methods Totally 112 poststroke anxiety patients and 246 non-anxious stroke controls were included and completed Hamilton Anxiety Rating Scale.DNA was extracted from blood and genotyped for the two polymorphisms of THP2 gene by polymorphism chain reaction.Results The G allele of rs4570625 was associated with the increased risk of poststroke anxiety.Both the GG genotype and G allele were observed to be significantly higher in female case than in control.No significant difference in genotype and allele firequencies of the rs4565946 was found among the groups.Patients with the G-C haplotype had significantly increased the risk of poststroke anxiety compared to controls.Conclusions Our findings suggest that these ftmctional polymorphisms in TPH2 gene may be involved in development of poststroke anxiety.

Bronchoalveolar Lavage Fluid ; Humans ; Interleukin-1beta ; metabolism ; Macrophages, Alveolar ; cytology ; drug effects ; metabolism ; Silicon Dioxide ; adverse effects ; Silicosis ; metabolism

Objective Human sperm hyaluronidase in the acrosome is a key enzyme during fertilization.To study and establish a modified substrate film method to improve the diagnotic and treatment of male infertility and investigate the influence of sperm hyaluronidase on male fertility.Methods According to the biochemical feature of sperm hyaluronidase that can dissolve the hyaluronic acid in matrixes,the modified sodium hyaluronate-Gelatin membrane was used as substrate to demonstrate the sperm hyaluronidase activity by incubation and staining.70 human semen samples were selected,categorized and detected for hyaluronidase activity according to the resuh of routine clinical semen examination.The average sperm hyaluronidase activity was statistically anyliezed between the fertile group and infertile group.Results Under general light microscope,the clear and bright digestion circle around the sperm head can be observed against the background of deep purple-blue stained substrate at the positive-reaction area.The amounts of positive reaction and the diameter of the bright circle are positively related to the activity of sperm hyaluronidase.The average hyaluronidase reaction rate and diameter of normal fertile group were 70.84% and 78.17 ?m;What about the infertile group A were 60.02% and 76.92 ?m;The infertile group B were 29.11% and 8.22 ?m; There was statistical difference of HYD activity between the infertile group.B and the fertile group(P

OBJECTIVEThis study compared two newborn Cytomegalovirus (CMV) IgG antibody screening ELISA kits and evaluated the detection effectiveness of Abnova kit.

METHODSCMV IgG antibodies were detected by both SeraQuest and Abnova kits from dried blood spot (DBS) samples of 488 newborn heel sticks. The detection abilities of these two kits were compared in different sample dilution concentrations. Relative detection effectiveness of the Abnova kit was defined by statistical method using the SeraQuest kit as a point of comparison.

RESULTCompared to the SeraQuest screening test kit, the Abnova kit revealed a sensitivity of 98.9%, specificity of 78.6%, positive predictive value of 99.3%, negative predictive value of 68.8%, and the coincidence rate for these two screening test kits at 98.3%. The consistency check of both kits based on interpretation of the kappa statistic was relatively good. For the Abnova kit, the "area under the ROC curve" was 0.887, which indicates moderate accuracy.

CONCLUSIONAbnova kit can be applied to newborn screening for congenital CMV infections. However, repeating the test for ambiguous results is suggested to increase the specificity and negative predictive value.

Antibodies, Viral ; blood ; Cytomegalovirus ; immunology ; isolation & purification ; Cytomegalovirus Infections ; blood ; diagnosis ; virology ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Female ; Humans ; Immunoglobulin G ; blood ; Infant ; Infant, Newborn ; Male ; Neonatal Screening ; methods ; Reagent Kits, Diagnostic