The optimum condition of shaking-flask p roducing enzyme were the tempe rature 26℃,initial pH 6 4,fermentation period 19 hours,medium volume 15mL m e dium/300mL Flask.soymilk-clotting enzyme was obtained from ammonium sulfate p r ecipitation.The optimum temperature and pH for the soymilk-clotting activity wa s 70℃and 5 8.The enzyme was easy to lose activity in acid or alkaline circumst a nce.About 60% of the original activity remained after 1 hour at 60℃.Ca 2+ ,Fe 2+ , Mg 2+ ,Na +increased the clotting activity,whereas Zn 2+ ,Al 3+ ca use inhibition.

Adolescent ; Bacteria ; isolation & purification ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Respiratory Tract Infections ; microbiology

OBJECTIVETo explore the diagnostic values of vestibular autorotation test (VAT) for patients with vertebrobasilar insufficiency (VBI).

METHODSVAT and videonystagmography ( VNG) were performed on 73 patients with VBI and 48 patients with peripheral vestibular lesions (contrast group). Parameters analyzed included Gain, phase and asymmetry of VAT, as well as the canal paresis (CP) of caloric test and results of optokinetic-pursuit tests in VNG. Positive result of the test could be defined if anyone of the parameters was abnormal.

RESULTSFor VAT test, Gain was enhanced in VBI group and was reduced in contrast group. In VBI group and contrast group, Gain enhanced showed in 47 (64.4%) cases and 5 (10.4%) cases, respectively (chi2 = 31.19, P < 0.01). Simultaneity, Gain reduced in 11 cases (15.5%) and 22 cases (45.8%), respectively (chi2 = 13.82, P < 0.01). But there was no statistics significant for results of the parameters of phase, asymmetry and integration between two groups. For VNG test, results with optokinetic-pursuit tests were more abnormal in VBI group than that in contrast group, which showed central lesions characteristics. Forty-four cases (60.3%) in VBI group and 10 cases (20.8%) in control group showed central lesions results with optokinetic-pursuit tests and visual fixation test (chi2 = 15.89, P < 0.01). Unilateral or bilateral CP showed in 33 cases (68.6%) in control group and 51 cases (69.9%) in VBI group with caloric test.

CONCLUSIONSGain of VAT is mostly enhanced in VBI group, and Gain as a main characteristic is reduced in patients with peripheral vestibular lesions. The Gain parameter is availability for assessing characteristics of vestibular lesions. Phase and asymmetry can be used to assess the vestibular function but can not indicate the characteristics of vestibular lesions.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Vertebrobasilar Insufficiency ; complications ; physiopathology ; Vertigo ; etiology ; physiopathology ; Vestibular Function Tests ; methods ; Young Adult

OBJECTIVETo explore the possibility of using an endovascular microcoil as a gene delivery system.

METHODSAnti-adenoviral monoclonal antibodies were covalently attached to the collagen-coated surface of platinum microcoil. These antibodies were used to tether adenovirus encoding green fluorescent protein (Ad-GFP). Cell culture studies with rat arterial smooth muscle cells (A10) assessed transduction on or near the coil. Platinum coils coated with Ad-GFP were implanted into the ligated common carotid artery (CCA) of adult rats in a model of arterial stasis and pressurization. After 7 days, CCA segments were harvested, and coils were removed for histopathology and GFP expression studies, while organs were evaluated by polymerase chain reaction to assess viral biodistribution.

RESULTSIn cell culture studies, GFP-positive smooth muscle cells were detected only on the platinum coil surface. After 7 days, GFP was detected on the harvested platinum coil and in the organizing thrombus within the CCA according to fluorescence microscopy and immunohistochemistry. Morphometric analyses revealed that (13.3 +/- 2.0)% of cells within the organized thrombus were transduced with Ad-GFP via the gene delivery system. Ad-GFP was not detectable by polymerase chain reaction in lung, liver, or kidney.

CONCLUSIONSGene delivery endovascular microcoils represents an interventional device-based gene therapy system that can serve as a suitable platform for either single or multiple gene therapy vectors.

Adenoviridae ; genetics ; immunology ; Aneurysm ; surgery ; Animals ; Antibodies, Viral ; chemistry ; metabolism ; Biological Availability ; Carotid Artery, Common ; drug effects ; metabolism ; surgery ; Cells, Cultured ; Drug Delivery Systems ; instrumentation ; Embolization, Therapeutic ; methods ; Endothelial Growth Factors ; therapeutic use ; Genetic Therapy ; instrumentation ; methods ; Genetic Vectors ; administration & dosage ; chemistry ; Green Fluorescent Proteins ; analysis ; Muscle, Smooth, Vascular ; cytology ; Platinum ; chemistry ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; therapeutic use ; Transduction, Genetic ; instrumentation ; methods

AIMTo produce poly (lactic-co-glycolic acid) (PLGA) microspheres, containing a staphylokinase variant (K35R, DGR) with reduced immunogenecity and antiplatelet aggregation activities, which allowed the preservation of protein stability during both particle processing and drug release.

METHODSDGR-loaded microspheres were fabricated using a double emulsion-solvent evaporation technique. The effects of preparative parameters, such as stirring rate, polymer concentration, and the excipients of both internal and external aqueous phase (W2), on DGR encapsulation efficiency and microsphere characteristics were investigated. In vitro and in vivo release of DGR were conducted and the cause for instability of DGR during release was also investigated.

RESULTSModerate ultrasonic treatment of aqueous DGR/dichloromethane mixtures caused approximately. Eighty four per cent DGR denaturation. However, the activity recovery of DGR almost amounted to 100% when 2% polyvinyl alcohol (PVA) was addled into the aqueous phase. It was found that NaCl in the external water phase significantly increased DGR encapsulation efficiency. Furthermore, NaCl in the external water phase played a role in determining size and surface morphology of microsphere. In vitro release test showed a burst release of DGR from microspheres, followed by sustained release of 50% total activity over 15 days. In vivo experiments showed that DGR released from microspheres sustained 5 days. Denaturation of DGR within microspheres might be resulted from acidic microclimate.

CONCLUSIONThe stability of DGR was effectively protected during microencapsulation and a relatively high encapsulation efficiency of DGR was obtained. PLGA microspheres could be an effective carrier for DGR.

Animals ; Area Under Curve ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Escherichia coli Proteins ; administration & dosage ; genetics ; pharmacokinetics ; Genetic Variation ; Lactic Acid ; Male ; Metalloendopeptidases ; administration & dosage ; genetics ; pharmacokinetics ; Microspheres ; Particle Size ; Polyglycolic Acid ; Polymers ; Rabbits

Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Middle Aged ; Papillomavirus Infections ; epidemiology ; Prevalence ; Risk Factors ; Uterine Cervical Neoplasms ; diagnosis ; epidemiology ; Young Adult

To construct Fv antibodies against H5N1 Avian influenza virus hemagglutinin,extracted mRNA from B lymphoblastoid cell lines secreting anti-HA antibodies was used and the VH and VL genes were amplified by RT-PCR and linked together by splicing overlap extension (SOE) with (Gly4 Ser)3 linker. The recombinant plasmid was then transformed to E. coli BL21(DE3) and sequence analysis indicated the total length of scFv was 714 bp and the expression of Fv was validated by PAGE and Western blot.
Animals ; Antibodies ; genetics ; metabolism ; pharmacology ; Birds ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation ; Hemagglutinins ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; metabolism ; Influenza A Virus, H5N1 Subtype ; drug effects ; immunology ; Influenza in Birds ; virology ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; immunology

OBJECTIVETo examine the association between autism spectrum disorder (ASD) and epilepsy in children.

METHODSA total of 190 children with ASD were enrolled. A self-designed questionnaire, Childhood Autism Rating Scale, and Autism Behavior Checklist were used to determine the association between ASD and epilepsy.

RESULTSAmong the 190 children with ASD, 20 (10.5%) had epileptic seizures and 12 (6.3%) were diagnosed with epilepsy. The rates of abnormal physical development and hearing disorders before the age of one year were significantly higher in ASD children with epileptic seizures than in those without epileptic seizures (P<0.05). The ASD children diagnosed with epilepsy and those receiving epilepsy treatment had a significantly increased rate of abnormal physical development before the age of one year (P<0.05). The ASD children with epileptic seizures had poorer sensory responses and behavioral competencies than those without epileptic seizures (P<0.05). Epilepsy treatment have a positive effect on behavioral competencies in ASD children (P<0.05).

CONCLUSIONSThere is a significant association between ASD and epilepsy in children. The possibility of the comorbidity between ASD and epilepsy may be assessed according to the status of growth and development before the age of one year, sensory responses and behavioral competencies, and the presence or absence of epileptic seizures.

Adolescent ; Autism Spectrum Disorder ; complications ; Child ; Child, Preschool ; Developmental Disabilities ; etiology ; Epilepsy ; complications ; Female ; Hearing Disorders ; etiology ; Humans ; Male

Objective To observe the preventive value of recombinant human granulocyte colony stimulating factor(rhG-CSF)in cancer patients after chemotherapy.Methods In the open study,enrolled 52 patients with previously untreated cancer and with normal bone marrow function were randomly divided into 2 matched groups,A and B group.Each patient received one cycle of chemotherapy.In the study cycle,the pa- tients received a single subcutaneous injection of rhG-CSF 150 ?g before 24 hours of chemotherapy and in control cycle the patients only received chemotherapy.Efficacy and safety parameters were monitored.Results The incidence rates of leukopenia in the 26 valuable study cycles and 26 valuable control cycles were 19.23 % and 53.85 %,There were significant lower than those of group B(P

Objective This study was conducted to determine the prognostic factors of cervical adenocarcinoma.Methods Between July 2008 and June 2013, 211 consecutive patients were treated because of International Federation of Gynecology and Obstetrics(FIGO) stage IA-IVB cervical adenocarcinoma.Overall survival was analyzed by the Kaplan-Meier method.Results One-year OS rate was 96.2%,and 3-year OS rate was 88.6%,and 5-year OS rate was 74.7%.Patients'age,FIGO stage,postoperative clinicopathological factors such as depth of stromal invasion,number of pelvic lymph node metastasis and para-aortic lymph node metastasis,were analyzed to discriminate their prognostic role.In the univariate analysis, age,FIGO stge,deep stromal invasion, lymph-vascular space invasion, number of pelvic lymph node metastasis,para-aortic lymph node metastasis were associated with poorer survival(P< 0.05).In the cox analysis, age (RR=4.940,95%CI:1.925-12.678),pelvic lymph node metastasis(RR=2.391,95%CI:1.311-4.360),para-aortic lymph node metastasis (RR=6.344,95%CI:1.007-39.990)and FIGO stage(Ⅲ vs I,RR=5.315,95%CI:2.760-12.203;Ⅳ vs I,RR=9.932,95%CI:2.905-18.846)were independent prognostic factors of invasive cervical adenocarcinoma.Conclusion Early detection and early treatment is very important for cervical adenocarcinoma patients.Give the patient suitable and maybe more aggressive adjuvant therapy according to surgical pathology is helpful for imporving adenocarcinoma treatment.