Comparative study of PCR effect of rapid PCR instrument and common PCR instrument / 中国法医学杂志

Objective To compare the PCR effect of rapid PCR instrument and common PCR instrument. Methods The concentration of 9947A standard was diluted to 0.1, 0.05, 0.025, 0.0125, 0.0063 ng/μL, 100 samples from each concentration group to establish PCR reaction system with Identifiler? Plus PCR Amplification Kit, 50 samples of them test with rapid PCR instrument (Speed cycler2 thermocycler), the other 50 samples test with common PCR instrument (9700 thermocycler). Detection of PCR product with 3500xL Genetic Analyser, the STR typing of both groups of each concentration group should be compared. Results The success rate of both thermocyclers have no significant difference (P>0.05); The success number of STR typing of common PCR instrument (13.7±1.0; 11.3±1.5) were higher than rapid PCR instrument (13.1±1.3; 9.9±1.9) when the concentration of 9947A were 0.0125, 0.0063ng/μL (P=0.029; P<0.001); The peak height of DNA typing map obtained from common PCR instrument (18931±4625;13437±3165; 5752±1344) were higher than rapid PCR instrument (16929±4034; 11815±4120; 4865±1401) when the concentration of 9947A were 0.1, 0.05, 0.025ng/μL (P=0.023; P=0.030; P=0.002). Conclusions The rapid PCR instrument could achieve the equal success rate to the common PCR instrument with less time, which revealed that the rapid PCR instrument was suitable for application in practical cases; The quality of STR typing from common PCR instrument may be more higher.

Comparative study of DNA extration of bloodstain on the iflter paper with four methods of solid phase adsorption / 中国法医学杂志

Objective To discuss the effect of DNA extraction of bloodstain on the filter paper with four methods of solid phase absorption.Methods 180 bloodstain samples on the iflter paper, each one contains 1 microlitre anticoagulation peripheral venous blood, divided into 4 groups with 45 samples, respectively. All samples were treated with four methods of solid phase absorption, i.e. DNA IQ? System, D-shield sensitive DNA Extraction Kit, High efficiency Silica Bead DNA Extraction Kit and Conventional silica bead method. The concentration of DNA and the results of STR typing of four groups were compared each other.Results The concentration of DNA was 3.764±1.790μg/mL and 3.634±1.112μg/mL by using D-shield sensitive DNA Extraction Kit and High efifciency Silica Bead DNA Extraction Kit, respectively. However, the concentration of DNA by using Conventional silica bead method group (3.350±1.250) was not signiifcantly different from each other (P<0.05), while the concentration of DNA extracted with above three methods were higher than by using DNA IQ? System (1.864±1.207)(P<0.001); Signiifcant differences of peak height existed between DNA IQ? System and other three methods (P<0.001); As the same time, the peak height of samples by using High efficiency Silica Bead DNA Extraction Kit and Conventional silica bead method were signiifcantly different from D-shield sensitive DNA Extraction Kit (P<0.01).Conclusions The DNA extracted in bloodstain on the iflter paper by using D-shield sensitive DNA Extraction Kit, High efifciency Silica Bead DNA Extraction Kit and Conventional silica bead method was more than DNA IQ? System. Meanwhile, the quality of DNA using High efifciency Silica Bead DNA Extraction Kit and Conventional silica bead method may be higher.

Design of absorbable Bondi of dura / 中国医疗器械杂志

A sort of absorbable Bondi of dura, whose main body is glue capsule, to compensate the deficiency of previous craniotomy, which easily causes delayed epidural hematoma. This device will help conglutinate dura to skull plate tightly, to stop bleeding and other purposes.

Association study between the genetic polymorphism of 15 STR loci and the crime of rape / 中华行为医学与脑科学杂志

Objective To investigate the relationship between rapists and related allele genes based on the analysis of 15 short tandem repeats (STRs) loci genetic polymorphism. Methods The method of Genome-wide scan was being used. Buccal swab samples of 129 rapists and 156 random populations were collected and PCR compound amplification was carried out with the aid of AmpFISTR Identifiler system. Then the products were subjected to electrophoresis and gene detection with AB13100 type gene analysis system so as to calculate and compare the alleles of 15 STRs gene frequency in the two groups. Results All the 15 STRs loci allele gene frequency in rapists and random population was found to coincide with Hardy-Weinberg law(P>0. 05). Allele 28 of D21S11 (rapists: 1.55% ,control group:5. 13%) ,allele22 of FGA(rapists:24.03% ,control group:16.99%),allele23 of FGA(rapists: 17.05% ,control group:26.28%) ,allele 10 of TH01(rapists:1.16% ,control group:4.17%) ,allele 8 of TPOX(rapists:55.77% ,control group:63.77%),allele 12 of TPOX(rapists:4.26% ,control group: 1.28%) were different between the two groups (P< 0.05) .while it is no differ significantly in other STRs loci allele gene(P >0.05). Conclusion Allele 28 of D21 S11,allele 22 and 23 of FGA, allele 10 of TH01, allele 8 and 12 of TPOX may be associated with the violent crime of rape. It is suggested that there are existing sensitive or resistance genes about the violent crime of rape in chromosome 2,4,11,21.

Immunomodulatory effect and neuronal-like cells differentiation of bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:As a kind of undifferentiated precursor cells,the phenotypic differentiation of bone marrow mesenchymal stem cells (BMSCs) remains immaturity,thus it presents weak rejection following transplantation.However,the in vitro directional differentiation of BMSCs into neuronal cells is easy affected by various factors.OBJECTIVE:To observe the immunomodulatory effect and the potential of BMSCs differentiate into neuronal-like cells.DESIGN,TIME AND SETTING:A contrast observation was conducted at the Department of Cytology,Third People's Hospital of Wuxi from January 2008 to March 2009.MATERIALS:Bone marrow was harvested from chips of cancellous or ilium bone dudng hip joint surgery.METHODS:Firstly,the BMSCs were separated and cultured to establish mixed lymphocyte reaction (MLR) system.Secondly,2 samples of peripheral blood mononuclear cells (1×10~5/well) were added into 96-well plate,and then BMSCs treated by mitomycin were added according to different ratios (BMSCs/peripheral blood monouclear cells).At the end,the cells were cultured as follows:Method 1:DMEM+10% fetal calf serum+1 μmol/L RA +20 μg/L basic fibroblast growth factor+20 μg/L epidermal growth factor.Method 2:DMEM+2% dimethyl sulfoxide +100 μmol/L butylated hydroxyanisole.MAIN OUTCOME MEASURES:The growth rate of lymphocyte was measured by ~3H-Thymidine,and the effect of BMSCs on lymphocyte proliferation was observed.Additionally,the differentiation potential of BMSCs into neuronel cells was determined by immunofluorescenca and immunohistochemistrical staining.RESULTS:①The BMSCs inhibited lymphocyte proliferation in MLR system and the influence on proliferation of lymphocyte was direct related to ratio of BMSCs.②Under a light microscope,cytoplasm of BMScs were shrinkd,which presented typical perikaryon morphology at hour 2 after culture with method 1.The majority of BMSCs were formed neuronal-like cells without number changes at hours 3-5,which turned to be dipolar or multipolar neuronal shapes at day 3.There were 60%-70% neuronspecific enolase,45%-50% glial fibrillary acidic protein were positive expressed.However,the positive rate of nidogen was decreased 3.4%.Cells cultured with method 2 became smaller after 2 hours,formed dipolar or multipolar body cells,and most of cells were died after 48 hours.The 40%-50% neuronspecific enolase,35%-40% glial fibrillary acidic protein was found to be positive.The positive rate of nidogen was temporary increased to 63% at hour 2 after culture；however,it was decreased to 1.6% after 48 hours.CONCLUSION:BMSCs can differentiate into neuronal-like cells,as well as inhibit lymphocyte proliferation in MLR system,which possess down regulation effect on alloimmunity-reaction.

Electrochemical Properties of Sodium Selenite on a Bionic Membrane of Gold Electrode Modified by L-Cysteine Self-assembled Monolayer / 分析化学

The cyclic voltammetric behaviors of sodium selenite on the gold electrode modified by L-cysteine self-assembled monolayer (L-Syc,Au/SAMs) has been investigated in 0.05 mol/L H_2SO_4 solutions with sodium selenite. Sensitive anodic stripping current of selenium (Se) with the peak potential at 0.99 V (vs. SCE) has been observed. In comparison with the electrochemical properties of sodium selenite on the modified electrode with those on the naked gold electrode, it was found that selenite was reduced to selenium by the redox interaction with sulfur on the monolayer and the monolayer catalytically oxidizes Se as the electrode was positively scanned. According to the electrochemical properties of sodium selenite both on the naked and modified gold electrode, the Se deposition mechanism and catalytic mechanism as well as bionic membrane model were proposed for the bio-utilization of selenite through nano scale selenium.

Improved efficacy of P277 fused to heat shock protein 65 of Mycobacterium tuberculosis against diabetes in nonobese diabetic mice / 生物工程学报

To improve the efficacy of peptide P277 in preventing autoimmune diabetes, heat shock protein 65 kD (HSP65) of Mycobacterium tuberculosis var. bovis was fused with linear polypeptide epitope of P277 and expressed as soluble protein in Escherichia coli. The fusion protein HSP65-P277 was purified by anion exchange column chromatography and then used to immunize prediabetic NOD mice with three ip inoculations in absence of adjuvants. Serum samples from the immunized mice were collected monthly and the concentration of blood glucose was measured. The study showed that administration of HSP65-P277 to NOD mice could prevent the development of diabetes more efficiently than the peptide P277 itself or HSP65. Fused to heat shock protein 65 of Mycobacterium tuberculosis could improve the efficacy of diabetes prevention of P277 in nonobese diabetic mice. The results suggest the fusion protein of HSP65-P277 would be useful for treating insulin-dependent diabetes mellitus.

Natural History of Intracranial Aneurysms / 国际脑血管病杂志

Intracranial aneurysms are a severe cerebrovascular disease.Its incidences ranged from 2% to 4% in general population.With the population aging and the development of imaging technique,the prevalence of intracranial aneurysm is increasing year by year.The research of natural course of aneurysm has important significance for neurosurgeons to evaluate the validity of therapeutic measures.There are significant differences between the patients with aneurysm who have had subarachnoid hemorrhage and those who have not.The rates of ruptured intracranial aneurysms per year ranged from 0.066% to 2%.The risk factors that affect ruptured intracranial aneurysms including the size and place of aneurysm,whether it is multiple aneurysms or not,growth rate,and self-factors of patients,such as age,sex,and history of hypertension and smoking,etc.

Comparison of different genesis of olfactory ensheathing cell transplantation for intracerebral hemorrhage / 中国组织工程研究

0.05). CONCLUSION: There are no significant differences in cell characteristics and transplantation outcome using OB-OEC and OM-OEC transplantation for repairing neurological function.

Effect of Displaying P277 Peptide on Surface of L-asparaginase on Its Antigenicity and Efficacy of Autoimmune Diabetes Prevention in NOD Mice / 中国生物化学与分子生物学报

The recombinant chimeric enzyme of AnsB-TTP-P277 comprising L-asparaginase, a tetanus toxin peptide spacer and P277 was expressed as a soluble protein in Escherichia coli. The purified chimeric enzyme exhibited primary activity of the native asparaginase. Prediabetic NOD mice immunized with the chimeric enzyme could induce specific antibodies against P277 and the specificity of anti-P277 antibodies was verified by Western blot assay. The study showed that displaying the P277 epitope on the surface of asparaginase could effectively overcome the weak antigenicity of the P277 epitope and evoke a strong P277-specific immune response in mice. Moreover, the concentration of blood glucose was measured by an automated analyzer.Histochemical analysis of mice pancreas tissues showed that the administration of the chimeric enzyme to NOD mice could prevent the development of diabetes more efficiently than the peptide P277 itself.

Intranasal vaccination with p277 tandem repeat sequences carried by Hsp65 prevented type 1 diabetes in NOD mice / 中国临床药理学与治疗学

AIM: To improve the prevent efficacy of peptide p277 in autoimmune diabetes. METHODS: The recombinant expression plasmid pET28-Hsp65-6×p277 was constructed by inserting 6×p277 which were amplified by PCR into the vector pET28-Hsp65. The plasmid pET28-Hsp65-6×p277 was transformed into E.coli BL21 (DE3) and the fusion protein (Hsp65-6×p277) was expressed effectively as soluble protein after inducing by lactose. The fusion protein was purified and then used to immunize 4-week old female NOD mice with three times of i.n. inoculations in the absence of adjuvants. Serum samples from the immunized mice were collected at monthly interval. The concentrations of blood glucose and antibodies were measured by automatic analyzer. RESULTS: Administration with the Hsp65-6×p277 to NOD mice could prevent the development of diabetes. CONCLUSION: The fusion protein Hsp65-6×p277 might be further developed to a vaccine against insulin-dependent diabetes mellitus.

Screening for antiviral fractions from ethanol extract of Euphorbia Kansui Liou / 中成药

Objective: To elucidate antiviral fractions from ethanol extract of Euphorbia kansui .Methods: Initial separation of ethanol extract from Euphorbia kansui was performed by column chromatography. The test mice were infected with flu virus mouse pneumonia adaptive strain (FM 1) and then treated with separated fractions intragastrally for determining lung index inhibition rate. The T lymphocyte proliferation enhancement was observed by the dosimetry of 3 H TdR infiltrated into DNA of the cell.Result: Initial separation of the extract afforded to 15 fractions. Fractions 8～15 with the elution volume from 29500 to 95000mL were highly effective to the therapy of the mice pneumonia. The enhancement to the lymphocyte proliferation of the 8 fractions under low concentration were 2～3 times higher than that of the control.Conclusion: Fractions 5 and 8～15 exhibited strong in vivo antiviral activity to the test virus, which might be realized by stimulating lymphocyte proliferation, enhancing the capacity of killing the cells infected by the virus.