An acidic xylanase gene,named xyl3,was cloned from the genomic library of enviromental microbes constructed by using shotgun cloning strategy,and submitted to GeneBank with accession number of gb:AY300805.BLAST analysis indicated that the gene xyl3 has low similarity with other xylanase genes and the encoded xylanase,sorted as Glycosyl hydrolases family 10,has 77% similarity with the intra-cellular xylanase from Geobacillus stearothermophilus at amino acid level.Treated with T4 DNA polymerase,the gene xyl3 was ligated with the linearized Pichia pastoris expression vector pHBM905 produced by digestion of restriction endonuclease CpoI and NotI to generate the recombinant plasmid pHBM706.Then the plasmid pHBM706,digested by restriction endonuclease SalI,was transformed into P.pastoris GS115 to obtain the recombinant P.pastoris GS115(pHBM706),which was induced to produce the recombinant xylanase with 0.5% methanol at 28℃.At the 36th hours of induction,the porduced crude enzyme was detected to reach the higest enzyme activity of 0.177 IU/mL.The optimal pH and temperature of the enzyme activity is 5.5 and 50℃ respectively.

OBJECTIVETo study the levels of CD4+ CD25+ Foxp3+ regulatory T cells (Tregs) in the peripheral blood of children with epilepsy and the roles of Tregs in the pathogenesis of epilepsy.

METHODSForty-one children with epilepsy and thirty-eight healthy children were enrolled. The percentage of CD4+ CD25+ Foxp3+ Tregs in CD4+ T cells and the percentages of CD4+ T cells, CD8+ T cells, nature killer (NK) cells and B cells in lymphocytes were evaluated by flow cytometry.

RESULTSThe percentages of peripheral blood CD4+ CD25+ Foxp3+ Tregs and CD4+ T cells and the ratio of CD4+ T cells to CD8+ T cells in epileptic children were (2.4±0.5)%, (35±5)% and 1.32±0.24 respectively, which were significantly lower than those in healthy children ［(6.1±1.2)%, (38±4)% and 1.60±0.24 respectively; P<0.05］. In contrast, the percentages of CD8+ T cells, NK cells and B cells in lymphocytes in epileptic children were significantly higher than those in healthy children［(27±3)% vs (24±3)%, (11.1±5.1)% vs (8.5±1.9)% and (24±9)% vs (16±5)% respectively; P<0.05］.

CONCLUSIONSThe abnormal percentage of CD4+ CD25+ Foxp3+ Tregs in the peripheral blood may be involved in the pathogenesis of epilepsy.

CD4-CD8 Ratio ; Child ; Child, Preschool ; Epilepsy ; immunology ; Female ; Humans ; Infant ; Killer Cells, Natural ; immunology ; Male ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo investigate the effect of Xianxiong decoction on the mice with acute lung injury induced by lipopolysaccharide.

METHODEighty female ICR mice were randomly divided into 8 groups: model group, Xianxiong decoction group, Daxianxiong decoction group, Xianxiong decoction group without Kansui Radix group, Xianxiong decoction group without Glycyrrhizae Radix et Rhizoma group, Glycyrrhizae Radix et Rhizoma and Kansui Radix group, normal group and control group. Animals of each group, except normal group, were undertaken intraperitoneal injection and intranasal inhalation of lipopolysaccharide (LPS) on day 1, 2, 3 to establish acute lung injury (ALI) model. 30 min after modeling, 0.2 mL corresponding drugs were administrated to each mice, dexam ethasone and normal saline were given to the mice of control group and normal group respectively. White blood cell in blood, neutrophil percentage of blood and bronchoalveolar lavage fluid (BALF) supernatant, the ratio of wet and dry lung tissue ( W/D), histopathological changes of lung tissue were estimated. Sixty ICR mice were randomly divided into normal, model, control, high, middle and low dose Xianxiong decoction groups and were modeled in the same way. ELISA was applied to detect the level of NF-kappaB, TNF-alpha and IL-6 in BALF, PCR for NF-kappaB and TNF-alpha mRNA in lung tissue, and Western blot for NF-kappaB and TNF-alpha. Half of 20 ICR mice were administrated with Xianxiong decoction of its maximum tolerant normal saline.

RESULTCompared with model group, the number of WBC in blood of Xianxiong decoction group mice decreased (P < 0.01), percentage of neutrophils in both blood and BALF decreased as well (P < 0.01, P < 0.05); it also significantly reduced the ratio of W/D (P < 0.01); and found the alveolar wall, the number of inflammatory cells infiltrating improved, compared with model group. Xianxiong decoction reduced the level of NF-kappaB, TNF-alpha and IL-6 in BALF (P < 0.01, P < 0.01, P < 0.05); its high and low dose groups only found TNF-alpha level declined. Five mice died 24 h after administration of Xianxiong decoction which indicated its toxicity when other influential factors were considered.

CONCLUSIONXianxiong decoction is effective on the ALI mice induced by LPS, but it is of toxicity at 3 g x mL(-1).

Acute Lung Injury ; drug therapy ; genetics ; metabolism ; pathology ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Interleukin-6 ; genetics ; metabolism ; Lipopolysaccharides ; adverse effects ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred ICR ; NF-kappa B ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is an autosomal recessive disease caused by SLC25A13 gene mutations, and is characterized by delayed jaundice clearance, liver dysfunction, and elevated aminoacidemia. The confirmed diagnosis depends on gene analysis. Citrin deficiency is one of the important causes of neonatal intrahepatic cholestasis in China. Recently more and more researches about NICCD were reported. The paper summarized the epidemiology, pathogenesis, clinical characteristics, and progresses in diagnosis and treatment of NICCD.

0.05).The setting time of hydroxyaptite and glass ions cements with Co-F were longer but there was little effect on zinc phosphate cements. Conclusion The Co-F agent added to dental cement can not only improve the compressive strength but also contin- ually release fluoride.

ObjectiveTo retrospectively analyze the distribution of artery involvement above the aortic arch and differences incidence of cerebral ischemic in patients with Takayasu’s arteritis (TA) by color Doppler flow imaging (CDFI).MethodsA total of 84 cases (aged from 11 to 70 years old, median age 39 years old) continuous observed from August 2005 to June 2013 were included in this study. All patients were detected by CDFI and confirmed by digital subtraction angiography (DSA) and clinical examination. The distribution of the involved arteries were analyzed.ResultsA total of 924 vessels (84 patients) was detected, 352 (38.1%) arteries involved inflammatory lesions. The prevalence rate of female was higher than male (92.9% and 7.1%) significantly. The symptoms of cerebral ischemic included dizziness, syncope, TIA, amaurosis etc. The involvement rate of common carotid artery (CCA) was the highest (94.0%), followed by the subclavian arterythe (SA) (82.1%). The detecting rate of bilateral CCA was higher than that of bilateral SA, 75% and 50% (χ2=15.732,P=0.003), respectively. The rate of combined lesions of CCA and SA were higher in left (64.3%) than the right 56% (χ2=12.777,P=0.001). Most patients had multiple vascular (≥ 3
vessels) diseases (69.1%). No significant difference was found in the incidence of cerebral ischemia symptoms in patients with bilateral CCA or SA lesions (74.6% with78.6%) (χ2=0.219,P=0.411). No significant difference was found inthe patients with left or right CCA combined with SA lesions (76.5% with 76.4% ,χ2=1.230, P=0.602) also. The incidence of cerebral ischemia for greater than 3 vessel disease was significantly higher than less vessels (69.1% with 30.9%) (χ2=6.674,P=0.010).ConclusionThe study in the distribution of aortic arch vascular lesion for the patients with Takayasu arteritis by CDFI is valuable for the early detection and treatment of vascular lesions, and decresing the incidence of ischemic cerebrovascular disease.

Objective To study the effects of 50 Hz magnetic fields on the in vitro proliferation of human osteosareoma cell line MG-63 with different cell densities.Methods Four different magnetic intensities(1 mT, 2 mT,3 roT,4 mT)were used to stimulate the cells,and the experiment was repeated with different cell densities. The method of MTT was employed to evaluate the level of proliferation.Results Fifty Hz magnetic fields signifi- cantly affected the level of proliferation of human osteosareoma cell line MG-63,and the 2 mT intensity exerted the greatest influence on it.The effects of the magnetic field differed with different cell densities.Conclusion The effect of 50 Hz magnetic fields on the in vitro proliferation of human osteosarcoma cell line MG-63 was not only relat- ed to the magnetic intensity,but also the cell density,

Over the past three decades, more and more antisense drugs have been approved for marketing or clinical trails. Antisense technology has become the focus of pharmaceutical research due to its unique advantages in treating diseases and strong clinical development potential. There is a big difference from traditional small molecule chemical drugs, and macromolecular protein biological drugs. Antisense drugs are a very independent drug form. Antisense drugs were initially used to treat diseases with single gene mutations, but recently they have gradually begun to be used for the treatment of common diseases. Rational antisense drug design is crucial for disease treatment based on genetics. This paper reviews the latest progress in the field of action mechanism, chemical modification and delivery strategy of antisense drugs, and analyzes the current intractable problems. It is believed that with the resolution of these problems, the research of antisense drugs can reach a new level.

OBJECTIVETo explore the mechanism of acupuncture in treatment of Parkinson's disease.

METHODSFifty Wistar rats were randomly divided into a normal control group, a sham-operation control group, a model group, a Fengfu-Taichong group and a Shuanggu Yitong group. The hemilateral rotation Parkinson's disease model was prepared by micro-injection of 6-hydroxyl-dopamine into the right striatum. Effects of acupuncture at "Fengfu" (GV 16), "Taichong" (LR 3) hy Shoanggu Yitong needling method on proliferation and differentiation of nerve stem cells in the Parkinson's disease model rat were compared. Proliferation and numher changes of the nerve stem cells in the suhstantia nigra and the striatum were investigated with immunohistochemical method, and the number of nerve stem cells transforming neurons were observed with immunohistochemical double-labeling method.

RESULTSIn the two acupuncture groups, the proliferative cells in the substantia nigra and thestriatum of the destroyed side increased significantly (P < 0.01), and the number of nerve stem cells and the number of transformed neurons in the destroyed side of the substantia nigra and the striatum increased significantly only in the Shuanggu Yitong group (P < 0.01).

CONCLUSIONShuanggu Yitong needling method can induce proliferation and differentiation of nerve stem cells in the substantia nigra and the striatum regions in the Parkinson's disease model rat.

Acupuncture Therapy ; methods ; Animals ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Corpus Striatum ; pathology ; Male ; Neurons ; cytology ; Parkinson Disease ; therapy ; Rats ; Rats, Wistar ; Stem Cells ; cytology ; Substantia Nigra ; pathology

AIMTo establish a method for determinate of the inhibitory activity of angiotensin-converting enzyme inhibitor captopril by high performance capillary electrophoresis.

METHODSThe characteristic absorptive wavelength of hippuric acid determined by ultraviolet spectrophotometer is 228 nm. The method employed a melted capillary column, 50 mmol.L-1 phosphoric acid (pH 8.3) buffer solution, inject pressure 4.8 kPa, inject time 3 s, separation voltage 20 kV and detection wavelength 228 nm.

RESULTSThe reactant and resultant was separated completed within 7 min. IC50 of captopril was 0.019 mumol.L-1. Captopril is a competitive inhibitor, which was proved by enzyme reaction dynamics.

CONCLUSIONThe method was shown to be accurate, simple and rapid and can be used for determination of the inhibitory activity of captopril.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Captopril ; pharmacology ; Electrophoresis, Capillary ; methods ; Hippurates ; analysis ; Peptidyl-Dipeptidase A ; metabolism