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.

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

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

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.

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

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.

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

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

OBJECTIVETo explore the difference in the efficacy and effect mechanism of subcortical ischemic vascular disease (SIVD) complicated with depression between acupuncture and medication.

METHODSSixty patients were randomized-into an acupuncture group and a medication group, 30 cases in each one. In the acupuncture group, acupuncture was applied to Baihui (GV 20), Shuigou (GV 26), Fengfu (GV 16), Fengchi (GB 20) and the others, once a day, 6 times a week. The treatment of 4 weeks made one session and totally 2 sessions were required. In the medication group, nimodipine 30 mg, three times a day and fluoxetine 20 mg, once a day were prescribed for oral administration, for 8 weeks totally. Before treatment, at the end of the 4th week and at the end of the 8th week of treatment, cerebral blood flow velocity (CBFV) and solubility CD40 ligand (sCD40L) were determined respectively. The scores in Montreal cognitive assessment (MoCA) and Hamilton' s depression scale (HAMD) were evaluated in the two groups. The efficacies on cognitive function and depression symptoms were compared in the patients between the two groups. Results Compared with the outcome before treatment, mean blood flow velocity (Vm) of middle cerebral artery (MCA), anterior cerebral artery (ACA) and posterior cerebral artery (PCA) was increased significantly at the end of the 4th week of treatment in the two groups (all P<0.05). At the end of the 8th week, Vm was increased much significantly (all P<0.01). The differences were not significant in comparison between the two groups (all P>0.05). Compared with the expression before treatment, sCD40L was reduced significantly after treatment in the patients of the two groups (all P<0.01), but the differ- ence was not significant between the two groups (all P>0.05). Compared with that before treatment, MoCA score was increased significantly after treatment in the two groups (all P<0.01), HAMD score was reduced sig- nificantly (all P<0.01), the differences were not significant in comparison between the two groups (all P>0.05). The total effective rate of cognitive improvement was 86.7% (26/30) in the acupuncture group and was 80.0% (24/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05). The total effective rate of the improvement in depression was 93.3% (21/30) in the acupuncture group and was 86.7% (26/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05).

CONCLUSIONAcupuncture could significantly increases CBFV and reduces serum sCD40L expressions in the patients of SIVD complicated with depression, and significantly improves cognitive function and relieves depression symptoms. The efficacy of it is similar to that of western medication. The increase of serum sCD40L expression is possibly involved in the occurrence and development of SIVD. Reducing sCD40L expression contributes to the alleviation of damage induced by cerebral ischemia and reperfusion.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Cerebral Infarction ; complications ; drug therapy ; therapy ; Depression ; drug therapy ; etiology ; therapy ; Female ; Fluoxetine ; administration & dosage ; Humans ; Male ; Middle Aged ; Nimodipine ; administration & dosage ; Treatment Outcome