Animals ; Apoptosis ; Hypoxia-Inducible Factor 1, alpha Subunit ; Hypoxia-Ischemia, Brain ; genetics ; pathology ; Neurons ; cytology ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; genetics

Manganese peroxidase (MnP), a crucial enzyme in lignin degradation, has wide potential applications in environmental protection. However, large-scale industrial application of this enzyme is limited due to several factors primarily related to cost and availability. Special attention has been paid to the production of MnP from inexpensive sources, such as lignocellulosic residues, using solid-state fermentation (SSF) systems. In the present study, a suitable SSF medium for the production of MnP by Schizophyllum sp. F17 from agro-industrial residues has been optimized. The mixed solid medium, comprising pine sawdust, rice straw, and soybean powder at a ratio of 0.52:0.15:0.33, conferred a maximum enzyme activity of 11.18 U/g on the sixth day of SSF. The results show that the use of wastes such as pine sawdust and rice straw makes the enzyme production more economical as well as helps solve environmental problems.
Culture Media ; Fermentation ; Industrial Microbiology ; methods ; Oryza ; Peroxidases ; biosynthesis ; Schizophyllum ; enzymology ; Wood

OBJECTIVETo establish a general method of focal cerebral ischemia model in different varieties of mice.

METHODEach group of healthy adult KM and C57BL/6 mice were randomly divided into control group (n = 10) and MCAO group (n = 10). The mice in MCAO group were applied in the preparation of the MCAO model by intraluminal occlusion using monofilament. Twenty-four hours after operation,the neurologic function was evaluated,middle cerebral artery blood flow was monitored and the infarction volume was calculated by TTC staining, to evaluate the reliability of the model.

RESULTIn the MCAO group, the base value of the cerebral blood flow down of KM and C57BL/6 mice respectively was (81.65 ± 4.59)%, (83.68 ± 6.25)%. The neurological deficit score respectively was (2.30 ± 0.82), (2.50 ± 0.80). TTC staining can clearly show the infarction area, and relatively stable, 24 hours of the survival rate of KM and C57BL/6 mice were 100% and 80% respectively.

CONCLUSIONThe key link is the optimization and improvement of monofilament, temperature, anesthesia and so on. The modified intraluminal occlusion of MCAO using monofilament is a kind of reliable and simple method to establish experimental cerebral ischemia model in mice.

Animals ; Blood Flow Velocity ; Brain ; blood supply ; pathology ; physiopathology ; Brain Ischemia ; complications ; physiopathology ; Cerebrovascular Circulation ; Disease Models, Animal ; Infarction, Middle Cerebral Artery ; complications ; physiopathology ; Male ; Mice, Inbred C57BL ; Middle Cerebral Artery ; pathology ; physiopathology ; surgery ; Nervous System Diseases ; etiology ; physiopathology ; Species Specificity

Objective To investigate the effect of PTEN gene in the mouse uterus during embryo implantation.Methods Real-time fluorescence quantitative PCR(FQ-PCR) was used to detect PTEN mRNA expressed in the endometria of the nonpregnant mice and the late pregnant mice(day 1,day 3,day 4,day 5 and day 7),with 20 mice sacrificed at each fixed day.Out of another 20 3-day pregnant mice,ten received PTEN antisense oligonucleotide at the horn of uterus and ten received normal saline to count the blastocysts at pregnant day 8.Results The PTEN mRNA/?-actin mRNA in pregnant mice was higher than that of nonpregnant mice,gradually hoisted as days passed by,and reached the highest at pregnant day 5.The number of blastocysts in the mice that received PTEN antisense oligonucleotide was fewer than that received normal saline.Conclusion PTEN persistently expresses in mouse endometria during the early pregnancy and maybe participate in the regulation process of mouse blastodyst implantation.

A rapid and sensitive method has been developed for the simultaneous determination of four selenium species Se(Ⅵ), Se(Ⅵ), selenomethionine, and Se-methylselenocysteine in Se-enriched yeast by liquid chromatography-hydride generation atomic fluorescence spectrometry (HPLC-HG-AFS). The isolation of the analytes from yeast samples was accomplished by proteaseⅩⅣ and trypsin enzymatic digestion. The target compounds were separated on a PRP-X100 anion exchange column and analyzed by HG-AFS. The mobile phase was 20 mmol/L (NH4)2HPO4. Good linearity was obtained for all the selenium species, with linear correlation coefficients higher than 0. 9996. The LODs of the four species were between 0. 5 and 5. 0 μg/kg. Average recoveries for the four analytes were in the ranges of 82 . 5%-101 . 2%, with intra-and inter-day RSD lower than 8. 6% and 14. 5, respectively. The proposed analytical method is simple, sensitive, with low operation cost, making it applicable for the determination of the selenium species in Se-enriched feeds.

Objective To explore the diagnostic value of anti-cyclic citrullinated peptide (CCP) 3.1 IgG/IgA antibody detected by enzyme-linked immunosorbent assay (ELISA) in patients with rheumatoid arthritis (RA).Methods The ELISA was used to measure the anti-CCP3.1 antibody in the serum of 169 RA patients,100 patients with other rheumatic diseases (including systemic lupus erythematosus,Sjogren's syndrome and osteoarthritis) and 72 healthy controls.The diagnostic value of CCP3.1 was assessed and compared with the second generation of anti-CCP IgG (CCP2) antibody,the correlations between anti-CCP3.1 antibody and the clinical and laboratory parameters were analyzed.Two-independent samples t test,chi-square test and Spearman's correlation were adopted for statistical analysis.Results ① The average cut-off concentration of anti-CCP3.1 antibody was (1122±1429) U/ml in RA,(13±14) U/ml in other rheumatic diseases and (6±5) U/ml in healthy controls.② The area under curve of ROC for anti-CCP3.1 antibody and anti-CCP2 antibody were 0.923 and 0.936 respectively.There was no difference between the sensitivity (82％ vs 79％)and specificity (97％ vs 99％) of anti-CCP3.1 antibody and anti-CCP2 antibody.The Kappa values between anti-CCP3.1 antibody and anti-CCP2 antibody was 0.763.③ We also found that anti-CCP3.1 antibody was positive in 20％(7/35) of anti-CCP2 antibody negative,43％(18/42) of RF negative,62％(47/76) of AKA negative,71％(49/69) of APF negative and 13％ of autoantibodies negative patients,indicated that antiCCP3.1 antibody had a potential value in the diagnosis of serum negative patients with RA.④ The presence of anti-CCP3.1 antibody was correlated with RF,HRF-IgG,APF,AKA,GPI and IgA (P＜0.05),except disease activity.Conclusion The sensitivity of anti-CCP3.1 antibody is slightly higher than anti-CCP2 antibody.The Anti-CCP3.1 antibody is a very valuable parameter for the diagnosis of RA,especially in serum negative patients.

Animals ; Animals, Newborn ; Brain ; metabolism ; pathology ; Caspase 3 ; Caspases ; metabolism ; DNA-Binding Proteins ; genetics ; Gene Expression ; Hypoxia, Brain ; genetics ; physiopathology ; Hypoxia-Inducible Factor 1 ; Hypoxia-Inducible Factor 1, alpha Subunit ; Immunohistochemistry ; Ischemic Preconditioning ; Nuclear Proteins ; genetics ; RNA ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors

Adenocarcinoma ; metabolism ; pathology ; Apoptosis ; drug effects ; Baculoviral IAP Repeat-Containing 3 Protein ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromones ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Morpholines ; pharmacology ; Ovarian Neoplasms ; metabolism ; pathology ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; Ubiquitin-Protein Ligases ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; metabolism

Objective To study the clinical characteristics of children with demyelinating disease.Methods Age of onset,presymptoms,clinical manifestations and auxiliary examinations of 51 children diagnosed as demyelinating disease were analyzed retrospectively.Results The onset age was from 3 months to 14 years,and the number of school age children was 38.Before illness,32 cases were relevant to infection,3 cases to vaccine inoculation.Most of children had acute courses.In the initial stage,32 cases with peripheral nervous demyelinating disease presented to paralysis of limbs;18 cases with central nervous demyelinating disease presented to visual disorder,somatasthenia,fever,convulsion and headache.The EMGs of children with peripheral nervous demyelinating disease showed nervous lesion.Among 18 children with central nervous demyelinating disease,17 cases showed abnomal signal on MRI or CT.Conclusions Children with demyelinating disease displays diversified clinical manifestations.By investigating case history combined with auxiliary exaninations,it is not difficult to make a correct diagnosis and its prognosis is good commonly.

Objective:To investigate the effect on the expression of Slug for the trasfection of miR-200c combined with the research on the ability of migration of breast cancer cell BT549.Methods:Chemically synthesized miR-200c mimic was trasfected into BT549 cells,which have high metastatic potential.The effect on the ability of migration of breast cancer cell BT549 for the transfection of miR-200c was analyzed by Transwell migration assay and Wound healing assay.The expression of Slug and E-cadherin mRNA was detected by Real-time PCR.The expression of Slug protein was detected by Western blot.Results:Transfection with miR-200c mimic significantly down-regulated the expression of Slug as compared with the control group (P<0.05).BT549 cell tranfected with miR-200c mimic had lower levels of migration capacity than cells in the control group (P<0.05).Conclusion:miR-200c inhibits Epithelial-mes-enchymal transition by suppressing Slug leading to down-regulation of migration capacity of breast cancer cell BT549.